the complete Symposium Program Book - Merial

Transcription

Merial-NIH National
Veterinary Scholars Symposium
July 28 – 31, 2016
Transdisciplinary
Approaches to
Health and
Wellness
TABLE OF CONTENTS
Welcome ........................................................................................................................... 2
Sponsors............................................................................................................................ 3
Schedule of Events............................................................................................................ 4
Speaker Biographies
Thursday Evening Reception and Welcome............................................................... 8
Dignitary Biographies............................................................................................ 9
Friday Speakers........................................................................................................ 16
Keynote Speaker................................................................................................ 17
AVMA/AVMF Young Investigator Award Finalists............................................... 18
Friday Feature Speaker...................................................................................... 19
Breakout Session A Speakers............................................................................ 20
Breakout Session B Speakers............................................................................ 23
Friday Evening Speakers.......................................................................................... 25
Friday Evening Featured Speaker...................................................................... 27
Saturday Speakers.................................................................................................... 28
Saturday Feature Speaker.................................................................................. 29
Merial Research Award Recipients..................................................................... 29
AVMA/AVMF Award Recipients.......................................................................... 30
Breakout Session C Speakers............................................................................ 32
Breakout Session D Speakers............................................................................ 34
First Floor Map................................................................................................................. 37
Second Floor Map............................................................................................................ 38
Third Floor Map................................................................................................................ 39
Abstracts Listed Alphabetically......................................................................................... 40
Abstract Index By Research Area.................................................................................... 57
Symposium Participants by School.................................................................................. 75
Young Investigator Award Finalist Abstracts.................................................................... 86
Complete Abstracts (listed alphabetically)....................................................................... 91
Late-Submission Abstracts............................................................................................. 310
College of Veterinary Medicine
Office of Research & Graduate Studies
1900 Coffey Road
Columbus, OH 43210
Phone (614) 292-9203
Fax (614) 292-7185
July 28, 2016
Dear 2016 Merial-NIH National Veterinary Scholars Symposium Attendees,
The entire Ohio State University community welcomes you to the heartland and Buckeye country. We
are delighted to host this year’s Veterinary Scholars Symposium taking place at the downtown
Columbus Hyatt Convention Center and the College of Veterinary Medicine campus at The Ohio State
University. This year’s conference is once again projected to have record attendance with over 600
attendees.
The theme of the 2016 symposium, Transdisciplinary Approaches to Health and Wellness, provides you
with opportunities to explore a number of research topics including Infectious Diseases, Translational
Oncology, and Regenerative Medicine through keynote and featured presentations, breakout sessions,
and poster sessions. We also include a special breakout session entitled Research Career Pathways:
Three Journeys. This year’s keynote presentation entitled Combatting Emerging Viruses: One Health
Approach will be presented by Dr. Ab Osterhaus who is currently establishing new One Health
Institutes in Utrecht, Netherlands and Hannover, Germany. We are also excited to welcome Dr. Jan
Ramer to present her experiences in leading mountain Gorilla conservation efforts in Africa.
We hope that during your visit you will make new friends, meet future colleagues and employers, and
be inspired to be part of veterinary medical research and discoveries that will impact the health and
well-being of animals and humans.
Thank you to the sponsors and donors who have made the Merial-NIH Veterinary Scholars Symposium
possible, the guest speakers and awardees who have helped us build an outstanding agenda, and the
many Ohio State staff, students and faculty who have contributed their time and energy to bring this
symposium to fruition. In particular, special thanks to Michele Morscher and Kathy Froilan, who have
worked tirelessly to ensure every detail of the symposium has been considered, refined, and
implemented.
We hope you have a wonderful time with us and look forward to passing the torch to the National
Institutes of Health, who with support from Michigan State University, will be hosting the 2017
symposium.
Sincerely,
Patrick Green, PhD
Associate Dean for Research and Graduate Studies
The Ohio State University
2
Michael Oglesbee, DVM, PhD
Professor and Chair, Veterinary Biosciences
SPONSORS
Merial LTD
National Institutes of Health
Elsevier
Burroughs Wellcome Fund
Association of American Veterinary Medical Colleges
American Veterinary Medical Association
American Veterinary Medical Foundation
Comprehensive Cancer Center/College of Medicine
Discovery Theme Initiative in Infectious Disease
Office of Research
Ohio Veterinary Medical Association
American College of Laboratory Animal Medicine
Howard Hughes Medical Institute
Huntington Bank
Morris Animal Foundation
Society for Toxicologic Pathology
Thank you again for your support of the
2016 Merial-NIH National Veterinary Scholars Symposium!
3
2016 Merial-NIH National Veterinary Scholars Symposium
“Transdisciplinary Approaches to Health and Wellness”
July 28 – 31, 2016
Symposium Schedule
THURSDAY, JULY 28th
11:00 am -7:00
pm
12:00-1:00 pm
1:00-4:00 pm
4:00-6:00 pm
6:30-9:00 pm
Participants arrive for registration
(posters will be able to be hung after 7:00 pm in the UNION)
T Directors LUNCH meeting – Manuel Moro – NIH
Colloquium of Veterinary Training – ALL T DIRECTORS
Dual DVM/PhD Programs Meeting (Xinbin Chen from UC
Davis and Michael Atchison from U Penn)
Welcome Reception (beverages and appetizers)
7:00 pm
Welcome Address and Other Dignitaries:
REGENCY SOUTH
FOYER
MARION
MARION
MARION
REGENCY BALLROOM
Co-Organizers:
Patrick Green, PhD
Michael Oglesbee, DVM, PhD
Chair, Department of Veterinary Biosciences
Bruce A. McPheron, PhD
Executive Vice President and Provost
Rustin Moore, DVM, PhD
Dean, College of Veterinary Medicine
Roberto Alva, DVM, PhD
Senior Director, Clinical Research and Development
Executive Director, Merial Veterinary Scholars Program
Fabian Kausche, MS, Dr med vet
Global Head of R&D, Merial Inc.
Susan Sholtis, MBA
Head of North America Business Operations, Merial Inc.
Ted Y. Mashima, DVM
Associate Executive Director for Academic & Research Affairs,
AAVMC
Ed Murphey, DVM, PhD
Assistant Director, Education and Research Division,
AVMA
Manuel Moro, DVM, MPH, PhD
Program Official, Office of Research Infrastructure, Division of
Program Coordination, National Institutes of Health
FRIDAY, JULY 29th
7:00-8:00 am
8:00-8:15 am
8:15-9:15 am
Breakfast Served
Groups A & B posters put up by 8:00 am
Session Introduction: Michael Oglesbee, DVM, PhD
KEYNOTE SPEAKER
(Sponsored by The Ohio State University
Discovery Theme Initiative in Infectious Disease)
Ab Osterhaus, DVM, PhD
(University of Veterinary Medicine, Hannover)
Title: “Combatting Emerging Viruses: One Health Approach”
FRANKLIN
REGENCY BALLROOM
9:15 – 10:45 am
AVMA/AVMF Young Investigators Award Finalists
Moderated by Harm HogenEsch, DVM, PhD
(Purdue University)
Finalists:
REGENCY BALLROOM
1. Elshafa Ahmed, The Ohio State University
2. Yung-Yi Mosley, Purdue University
3. Megan Niederwerder, Kansas State University
4. Maciej Parys, Michigan State University
5. Nora Springer, Cornell University
10:45-11:00 am Morning Break – refreshments provided – OUTSIDE REGENCY BALLROOM
11:00-11:10 am
11:10-12:00 pm
12:00-1:00 pm
1:00-2:30 pm
1:00 - 2:30 pm
Session Introduction: Patrick Green, PhD
FEATURE SPEAKER
(Sponsored by The Ohio State University Comprehensive
Cancer Center & College of Medicine)
Cheryl London, DVM, PhD
(The Ohio State University/Tufts University)
Title: “Leveraging Comparative Oncology to Maximize
Translational Outcomes”
Lunch Break – boxed lunches
BREAKOUT SESSIONS
SESSION A
Title: Research Career Pathways:Three
Journeys
Sponsor: Society of Toxicologic Pathology
Moderator: Krista La Perle, DVM, PhD
(The Ohio State University)
ROOM LOCATION-FAIRFIELD
Speaker: Krista La Perle, DVM, PhD
Title: “Comparative Pathology of Animal
Models, Collaborative Research and Core
Facility Development: One Pathologist’s
Research Hat Trick!”
Speaker: Sarah Tannehill-Gregg, DVM,
PhD (Takeda Pharmaceuticals, Boston, MA)
Title: “A Career in Industry as a Toxicologic
Pathologist: My Journey in Joining the “Dark
Side”; Twists, Turns, Forks in the Road and
Loving What I Do!”
Speaker: Joelle Fenger, DVM, PhD
Title: “Of Mice, Dogs and Men: A Career in
Comparative and Translational Oncology”
REGENCY BALLROOM
FRANKLIN
SESSION B
Title: Translational Oncology
Sponsor: The Ohio State University
Comprehensive Cancer Center
Moderator: Cheryl London, DVM, PhD
(The Ohio State University/Tufts University)
ROOM LOCATION-TAFT – C&D
Speaker: Matthew Breen, PhD
(North Carolina State University)
Title: “Comparative Oncology: What Can
Dogs (and Sea Lions) Teach Us About
Cancer?”
Speaker: Ryan Roberts, MD, PhD
(Nationwide Children’s Hospital, Columbus,
OH)
Title: “Enlisting Man’s Best Friend in the
Fight Against his Children’s Worst Enemy:
Seeking Common Cures”
2:30-2:45 pm Break
OUTSIDE OF UNION
Refreshments provided - Sponsored by Society of Toxicologic Pathology
2:45-4:15 pm
POSTER SESSION A
UNION
4:15-4:20 pm
4:20-5:50 pm
BREAK – PRESENTER SWITCH
POSTER SESSION B
UNION
ALL GROUPS A & B POSTERS DOWN BY 6:00 PM
6:00 pm
6:30 pm
7:30 pm
BUSES START CHARTERING TO THE OHIO STATE
COLLEGE OF VETERINARY MEDICINE FOR DINNER
Dinner Service begins
Welcome
Patrick Green, PhD
CVM COURTYARD
CVM COURTYARD
Rustin Moore, DVM, PhD
Dean, The Ohio State University College of Veterinary Medicine
Caroline Whitacre, PhD
Vice President Ohio State Office of Research
Brad Fenwick, DVM, PhD
Professor and Jefferson Science Fellow US Dept of State
Senior Vice President Global Strategic Alliances, Elsevier
FEATURE SPEAKER
(Friday Night’s Event sponsored by Elsevier)
Introduction by Mike Oglesbee, DVM, PhD
Jan Ramer, DVM
Director of Conservation Medicine
The Wilds, Cumberland, OH
9:00-11:00 pm
Title: “One Health/Conservation Medicine, Abroad and in
Your Backyard”
Buses will shuttle back to the Hyatt
SATURDAY, JULY 30th
7:00-8:00 am
8:00-8:10 am
8:10-9:00 am
9:00-9:50 am
Breakfast Served
ALL GROUPS C & D posters put up NO LATER THAN 8:00
am
Session Introduction: Alicia Bertone, DVM PhD
FEATURE SPEAKER
(Sponsored by the Ohio Veterinary Medical Association)
James Cook, DVM, PhD (University of Missouri)
Title: “One Health – One Medicine: How Veterinary Medicine
is Fixing People Too”
FRANKLIN
Merial Veterinary Research Scholar Award
Presented by Monica D. Figueiredo, DVM, PhD
Merial Director Lead Finding and External Innovation
REGENCY BALLROOM
Laura LoBuglio, Third Year Veterinary Student
University of Minnesota
Title: “Identification and Inhibition of Oxidative
Stressed-Induced Apoptosis Pathways in
Beta Cells in Diabetes”
Merial Veterinary Research Graduate Student Award
Presented by Diane Larsen, DVM, PhD
Merial Head, Pharma Development Projects
Victoria Baxter, Postdoctoral Fellow, Johns
Hopkins Bloomberg School of Public Health
Title: “The Role of Interferon Gamma in the
Immunopathogenesis and Control of Sindbis Virus
During Nonfatal Alphavirus Encephalomyelitis“
REGENCY BALLROOM
9:50 -10:05 am Morning Break – refreshments provided – REGENCY SOUTH FOYER
10:05-11:30 am
AVMA Excellence in Research Awards (2 AWARDS):
Presented by Harm HogenEsch, DVM, PhD
REGENCY BALLROOM
AVMF/Winn Feline Foundation Research Award:
Andrea Fascetti, VMD, PhD
Professor, University of California, Davis
AVMA Lifetime Excellence in Research Award:
Susan Stover, DVM, PhD
11:30 – 1:00 pm
1:00 – 2:30 pm
1:00 - 2:30 pm
FRANKLIN
LUNCH BREAK – boxed lunches
MERIAL (MVSP) DIRECTORS LUNCH MEETING AT THIS
TIME IN THE MADISON
MADISON -DIRECTORS
BREAKOUT SESSIONS
SESSION C
SESSION D
Title: Regenerative Medicine
Title: Infectious Diseases
Sponsor: Ohio Veterinary Medical
Sponsor: The Ohio State University
Discovery Theme Initiative
Association
Moderator: Alicia Bertone, DVM, PhD
Moderator: Michael Oglesbee, DVM, PhD
ROOM LOCATION – FAIRFIELD
ROOM LOCATION – TAFT – C&D
Speaker: Alicia Bertone, DVM, PhD
Title: “Hybrid Veterinary Clinical Trials in
Regenerative Medicine”
Speaker: Kurt Hankenson, DVM, PhD
(Michigan State University)
Title: “Developing Novel Therapies for Bone
Regeneration: A Stem Cell Biology”
Approach”
Speaker: Daniel Gallego Perez, PhD
Title: “Nanotechnology-Enabled
Regenerative Medicine”
2:30-2:45 pm BREAK – refreshments provided
Speaker: Michael Oglesbee, DVM, PhD
Title: “Fever and Virus Infection of the
Brain: Good, Bad and Ugly?”
Speaker: Larry Schlesinger, MD
Title: “Approaches to the Study of
Tuberculosis”
Speaker: Gabriel Hamer, PhD
(Texas A&M)
Title: “Evaluating the Relative Role of
Different Mosquito Species to the
Transmission of Viruses”
OUTSIDE UNION
2:45-4:15 pm
POSTER SESSION C
UNION
4:15-4:20 pm
4:20-5:50 pm
BREAK – PRESENTER SWITCH
POSTER SESSION D
UNION
ALL POSTERS DOWN BY 6:00 PM
6:30 PM
DINNER SERVICE BEGINS
REGENCY BALLROOM
7:30 pm
REGENCY BALLROOM
8:00-10:00 pm
AVMA/AVMF Young Investigator Award Results
Harm HogenEsch, DVM, PhD
Entertainment
REGENCY BALLROOM
ST
SUNDAY, JULY 31
7:00-8:30 am
ALL DAY
Breakfast Served
DEPART FOR AIRPORT, HEAD HOME
FRANKLIN
THURSDAY EVENING RECEPTION
AND WELCOME
JULY 28, 2016
6:30 – 9:00 pm
REGENCY BALLROOM
8
Patrick L. Green, PhD
Robert H. Rainier Chair in Industrial Veterinary Medicine and
Research
Patrick Green obtained his PhD in oncology from the University
of Wisconsin-Madison McArdle Laboratory for Cancer Research
under the mentorship of Drs. Rex Risser and Howard Temin
studying the molecular pathogenesis of an infectious cancer
virus, Abelson murine leukemia virus. As a postdoctoral
researcher and research scientist at UCLA and subsequently as an assistant professor at
Vanderbilt University, he pioneered some of the first human T cell leukemia virus (HTLV)
mutational gene studies in an infectious proviral clone and was instrumental in developing cell
culture approaches still used today to measure viral replication and cellular transformation.
In 1997, Dr. Green joined the faculty at The Ohio State University where members of his
laboratory continue to focus on cutting-edge questions in the HTLV field making use of a
combination of HTLV infectious molecular clones, natural target cells, and a reproducible rabbit
animal model of infection. His laboratory is one of a few worldwide that strive to study HTLV
gene structure/function and pathogenesis in this most biologically relevant context. Dr. Green is
currently Professor and Associate Dean for Research and Graduate Studies and Director of the
Center for Retrovirus Research, and holds the Robert H. Rainier Chair in Industrial Veterinary
Medicine and Research.
Dr. Green has over 25 years of research experience in the field of murine and human retroviral
pathogenesis. His research has been continuously funded by the National Institutes of Health
(NIH) since 1991 and he currently serves as principal investigator on multiple NIH grants
including an $8.7M NCI program project grant on retrovirus models of cancer. Dr. Green has
been appointed as a member of numerous NIH study sections and scientific panels and
currently serves as editor of AIDS Research and Human Retroviruses and as a member of the
editorial boards of Retrovirology and Journal of Virology. Dr Green also served for over 10
years as the leader of the Ohio State Comprehensive Cancer Center Viral Oncology Program.
He has been recognized with a number of honors that include the International Retrovirology
Association Award and the Pfizer Award for Research Excellence as well as designation as an
American Cancer Society Fellow, Leukemia Society of America Scholar, American Association
for the Advancement of Science (AAAS) Fellow, American Society for Microbiology Fellow, Ohio
State University Distinguished Scholar, and member of the Board of Trustees of the
Leukemia/Lymphoma Society.
Dr. Green has a documented commitment to mentoring and graduate training and strives to be
an energetic and enthusiastic teacher and role model. To date his track record of training
include three junior faculty, 14 graduated PhD students, and five postdoctoral fellows, many of
whom have received awards for their research and subsequently have gained important
positions at academic and government institutions and in the pharmaceutical industry. He has
also served as a dissertation committee member for over 55 doctoral or master’s students. Dr.
Green, along with his colleagues and trainees, has co-authored over 100 research publications,
reviews, and book chapters.
9
Michael J. Oglesbee, DVM, PhD, DACVP
Professor and Chair, Department of Veterinary Biosciences
Director, Ohio State CVM Summer Research Program
Faculty Lead, University Discovery Theme in Infectious Disease
Dr. Oglesbee is professor of virology and veterinary
neuropathology in the Department of Veterinary Biosciences,
College of Veterinary Medicine, The Ohio State University.
Scientific contributions have focused on host determinants of viral
neurovirulence, and he was named a fellow by the American
Association for the Advancement of Science in recognition of
distinguished contributions to our understanding of virus-heat
shock protein interactions related to infection, virulence, and
impact on innate and adaptive antiviral immunity. Dr. Oglesbee is chair of the Department of
Veterinary Biosciences, faculty director of the College Summer Research Program for veterinary
medical students at The Ohio State University, and principal investigator on an NIH training
grant (T35) in support of that program. Dr. Oglesbee leads the college’s signature program in
infectious disease, and is also the faculty lead on a university investment in infectious disease
involving the colleges of Veterinary Medicine, Medicine, Nursing, Food Agriculture and
Environmental Sciences, Public Health, Pharmacy, and Arts and Sciences, and the Food
Animal Health Research Program at the Ohio State Wooster Campus. Focus of the investment
is development of interdisciplinary collaborative research networks in the programmatic areas of
antimicrobial resistance mechanisms and drug discovery, treatment and prevention of virus
infection, host response to infection and microbial communities, and the ecology of pathogen
emergence and spread. The ultimate goal is to enhance our ability to control the spread and
severity of infectious disease through the creation and dissemination of knowledge, practices
and products.
10
Bruce A. McPheron, PhD
Executive Vice President and Provost
Bruce A. McPheron became Ohio State’s permanent chief
academic officer on June 1, 2016, after serving in an interim
capacity since December 2, 2015. As executive vice president and
provost, he oversees the administration, coordination, and
development
of
all
academic
functions
of
the
university. Previously, he had served as the vice president for
agricultural administration and dean of the College of Food,
Agricultural, and Environmental Sciences, beginning his
appointment in November, 2012. He joined The Ohio State
University after serving for three and one-half years as Dean of
Penn State University’s College of Agricultural Sciences. McPheron received his BS degree
from The Ohio State University and his MS and PhD degrees from the University of Illinois. He
served as the 2013-14 chair of the Policy Board of Directors of Agriculture for the Association of
Public and Land-Grant Universities. Currently, Dr. McPheron is a member of the Discovery
Theme executive team and leads the university’s Food Production and Security theme.
In his previous role, Dr. McPheron had leadership responsibility for over 2,100 faculty, staff and
student employees across the state and more than 3,700 graduate and undergraduate students
on the Columbus and Wooster campuses. His administrative leadership portfolio also included
the research and outreach arms of the college, including Ohio State University Extension and
the Ohio Agricultural Research and Development Center.
Since arriving at The Ohio State University, Dr. McPheron has initiated strategic planning or
review processes that will position the college for a successful and sustainable future. The
college has completed a Facilities Master Plan, Animal Facilities Re-Envisioning plan and the
Ohio State ATI Re-Envisioning initiative. The Vice President’s Conversation on the Future of
Extension, convened by Dr. McPheron, will provide a roadmap to successfully address critical
issues facing Ohioans by 2035. Recognizing the importance of water quality, Dr. McPheron has
been instrumental in building statewide collaborations, led by OSU faculty and staff, to address
this critical issue in Ohio. In 2014, he announced the creation of the Field to Faucet initiative that
is serving as a coordinating effort to address water quality in Ohio.
Dr. McPheron’s research expertise focused on the use of genetic tools to examine population
structure in pest insects. Molecular diagnostic methods for source identification of the
Mediterranean fruit fly developed in his laboratory have been adopted by USDA-APHIS and
several state and international agencies. He has taught a wide range of classes in entomology
and has traveled internationally sharing his expertise.
11
Rustin M. Moore, DVM, PhD, DACVS
Dean
Ruth Stanton Chair in Veterinary Medicine
Dr. Rustin M. Moore, the Ruth Stanton Chair in Veterinary
Medicine, officially became the 11th dean of the College of
Veterinary Medicine on Sept. 1, 2015.
A two-time graduate of the college, Dr. Moore has served Ohio
State as chair of the Department of Veterinary Clinical Sciences,
executive director of the Veterinary Medical Center, associate
dean for Clinical and Outreach Programs, and associate executive dean.
He has taught at all levels of the undergraduate, professional and graduate curricula, both at
Ohio State and at Louisiana State University (LSU), where he served on the LSU faculty from
1994 to 2006. He has served as an advisor, co-advisor or committee member for 21 doctoral or
master’s students as well as the clinical advisor for more than 25 interns and residents.
His clinical interests include equine lameness, surgery, and colic and its associated
complications, and his research has led to his being a principal or co-investigator on
approximately 120 funded grants. In addition, he has authored or co-authored more than 15
book chapters, and his work has been published in 120 peer- or editor-reviewed manuscripts
and 175 scientific abstracts. His service on editorial review boards includes the journal
Veterinary Surgery, and he has served as a manuscript reviewer for several additional
prestigious journals. He is also a frequently invited speaker at national and international equine
veterinary clinical, research and educational symposia.
In addition to extensive and substantial service to his department, college and the university, Dr.
Moore has contributed wide-ranging service and outreach efforts to his discipline, to the
community and beyond. For example, while at LSU, he assumed emergency leadership of a
large-scale rescue effort of nearly 500 horses and other animals during the aftermath of
Hurricanes Katrina and Rita. A national leader, he has served as president of the American
Association of Veterinary Clinicians, on the board of directors of the American Association of
Equine Practitioners, on the board of regents of the American College of Veterinary Surgeons
and as an equine health advisory board member for the Ohio Department of Agriculture, among
others.
In addition, among his numerous awards, he received the Pfizer Award for Research
Excellence, the School of Veterinary Medicine’s Distinguished Faculty, and the university’s
Distinguished Scholar Award and Lifetime Achievement Award. While at LSU, he received the
Lifetime Achievement Award from the International Equine Conference on Laminitis and
Diseases of the Foot and was recently inducted into West Virginia University’s Academy of
Distinguished Alumni.
A native of Spencer, WV, he earned a BS, summa cum laude, from West Virginia University and
a DVM, summa cum laude, and PhD from The Ohio State University. In addition, he is a
diplomate of the American College of Veterinary Surgeons.
12
Roberto Alva, DVM, MS, PhD
Senior Director Clinical R&D
Pharmaceutical and Biological Research & Development
Executive Director Merial Veterinary
Scholars Program
Merial Limited
Dr. Alva received his DVM degree from the National Autonomous
University of Mexico in Mexico City. Following graduation, he
worked as a swine practitioner for the Mexican federal
government. Dr. Alva moved to the United States where he
earned an MS in Animal Science and a PhD in Veterinary
Pathology from Iowa State University. He has also conducted
postdoctoral research at the University of Bern in Switzerland.
In 1982, Dr. Alva joined Merck and Co. Inc. to work primarily on the development of antiparasitic
drugs. He became a project leader and a pathologist for the company, developing a wide range
of pharmaceutical products. He later moved to Georgia with the same company to work on
clinical operations for North America and regulatory affairs for Latin America.
Currently, Dr. Alva leads a group of talented scientists at Merial that are responsible for
establishing the safety and efficacy of new pharmaceutical drugs and vaccines and for
developing new claims for existing products. He is the author or co-author of over 60 scientific
articles and abstracts. Dr. Alva leads the Merial Veterinary Scholars Program.
Fabian M. Kausche, MS, Dr med vet
Global Head of Merial Research & Development
Merial, a Sanofi Company
Duluth, GA
Fabian Kausche is the Chief Scientific Officer and Global Head
of Research and Development for Merial, an animal health
company focused on providing a comprehensive range of
solutions focused on disease prevention and health
management of animals that includes vaccines, parasiticides,
wellness products and therapeutic medicines.
Fabian leads a group of approximately 800 scientists and
support staff at sites around the world in advancing the scientific
and technical aspects of Merial research and development
programs. He also serves as a member of the Merial Executive
Committee and Merial Leadership Team.
Holding both German and United States citizenships, Fabian received a veterinary degree and a
Dr. med. vet. (PhD equivalent) from the Hanover Veterinary School in Germany, as well as a
Masters of Science degree from Iowa State University. Prior to joining Merial, he held a variety
of positions at Novartis in the animal health and human health organizations, including Global
Head of R&D for Novartis Animal Health and Senior VP and Global Head of R&D for the
Novartis over‐the‐counter human health business. Previously he spent time with Pharmacia &
Upjohn (later acquired by Pfizer). He is the author of numerous scientific publications, abstracts
and invited presentations. Having completed the Advanced Management Program in 2005, he is
also an alumnus of Harvard Business School.
13
Susan Sholtis, MBA
Head of North America Business Operations
Merial Inc.
Susan has over 25 years of experience including
both global and domestic assignments in sales,
marketing and general management. She started as
a pharmaceutical sales representative in 1991 in
Portland, Oregon. This started a career that took her
through several life sciences sectors including:
pharmaceutical, animal health and infant and children’s nutrition. Susan previously spent eight
years at Merial beginning in 1996 where she had global responsibility for managing two of the
largest brands in animal healthcare, FRONTLINE® and HEARTGARD®. She is known as the
architect behind the successful global launch of Merial’s FRONTLINE® Plus product, the number
one flea and tick product in the world. Following her initial experience at Merial, Susan held
various positions in executive level operations, business development, sales and marketing.
She moved to Mead Johnson Nutrition in 2006, where her career evolved from Director, US
Marketing, to General Manager, Northern Europe in 2007 followed by Vice President, Sales and
Marketing, Europe in 2009. During this time she was instrumental in leading the European
business through an IPO as the company separated from Bristol-Myers Squibb Co. Susan’s last
role at Mead Johnson Nutrition was as Head of Global Marketing. Susan returned to Merial in
2016 as Head of North America Business Operations. Susan graduated from Butler University
in Indianapolis, Indiana with a B.S. degree in Public & Corporate Communications. She
completed her MBA at Emory University in Atlanta, Georgia.
Ted Y. Mashima, DVM, DACZM, DACVPM
Associate Executive Director for Academic and Research Affairs
Association of American Veterinary Medical Colleges
Ted Y. Mashima is the Associate Executive Director for
Academic and Research Affairs for the Association of American
Veterinary Medical Colleges, a nonprofit organization that
coordinates the affairs of the veterinary colleges in the US and
other academic veterinary medical institutions internationally. He
previously served as the president and executive director of the
Asian & Pacific Islander American Scholarship Fund, associate
director for the Center for Public and Corporate Veterinary
Medicine (Virginia-Maryland College of Veterinary Medicine) and
the projects director for the National Association of Physicians for the Environment. He has a
BA (zoology) from the University of Hawaii at Manoa and a DVM from Colorado State
University. He completed an internship in zoological medicine at Kansas State University and a
residency in zoological medicine, wildlife emphasis, at North Carolina State University. He is
board certified in both zoological medicine and veterinary preventive medicine. In the past, he
was the vice president of the American College of Zoological Medicine, on the board of directors
for the Alliance of Veterinarians for the Environment, research associate with the Smithsonian
Institution’s National Zoological Park, and on the faculties of the Envirovet Summer Institute and
AQUAVET. He is the co-editor of The Rhino with Glue-On Shoes and Other Surprising True
Stories of Zoo Vets and Their Patients.
14
Ed Murphey, DVM, PhD, DACVS
Assistant Director, Education and Research Division
American Veterinary Medical Association
Dr. Erle Douglas "Ed" Murphey III joined the AVMA in 2012 as
an assistant director in the Education and Research Division. His
primary responsibilities are working as the staff liaison for the
AVMA Council on Research and the AVMA American Board of
Veterinary Specialties, and he has directed the development of
the AVMA Animal Health Studies Database, which went live in
June 2016. Dr. Murphey received his DVM from The Ohio State
University in 1989 and completed an internship in New
Hampshire in equine medicine and surgery immediately after. He
began his career working at a mixed animal practice for two years in Ohio before completing his
residency in large animal surgery at Purdue University. Afterward, Dr. Murphey earned a
doctorate in experimental pathology in 2000 from the University of Texas Medical Branch
(UTMB) and became a Diplomate of the American College of Veterinary Surgeons the same
year. He worked as a clinical instructor at the Washington State University and University of
Georgia colleges of veterinary medicine before returning to UTMB, first as a National Institutes
of Health postdoctoral research fellow in trauma and burns, later as an instructor, and, finally, as
an assistant professor in the Department of Anesthesiology. During this time, Dr. Murphey
performed critical-care medical research at the Shriners Burn Institute, at UTMB in Galveston,
specifically researching the effect of toll-like receptor agonists on subsequent innate immune
function in models of injury and sepsis. He has published in, and served as a reviewer for,
journals ranging from the American Journal of Veterinary Research to the Journal of
Immunology.
Manuel H. Moro, DVM, MPH, PhD
Program Official
Office of Research Infrastructure
Division of Program Coordination
National Institutes of Health (NIH)
Dr. Moro is a Program Official at the Division of Comparative
Medicine, Office of Research Infrastructure Programs, Division of
Program Coordination, Planning and Strategic Initiatives, Office of
the Director, National Institutes of Health. Dr. Moro manages a
grant portfolio that incudes biomedical training for veterinarians,
animal resource centers, research grants, and resource-related
research grants. Prior to joining the NIH, Dr. Moro was a faculty
member at the College of Veterinary Medicine at Kansas State
University in Manhattan, Kansas, where he taught epidemiology,
zoonoses and public health courses to veterinary students. His research studies focused on the
ecology and immunopathology of tick-borne diseases.
Dr. Moro received a Doctor of Veterinary Medicine degree from the College of Veterinary
Medicine at Universidad Nacional Mayor de San Marcos in Lima, Peru. He practiced veterinary
medicine in large animal species. Later, he received a Master of Public Health from the Johns
Hopkins School of Public Health in Baltimore, Maryland. He received his PhD degree in
Veterinary Microbiology and Preventive Medicine at the College of Veterinary Medicine at Iowa
State University in Ames, Iowa. He did his postdoctoral training at the Laboratory of Molecular
Microbiology, Immunology and Immunogenetics at the Mayo Clinic in Rochester, Minnesota.
15
FRIDAY SPEAKERS
JULY 29, 2016
16
A.D.M.E. (Ab) Osterhaus, DVM, PhD
Professor
University of Veterinary Medicine, Hannover
Keynote Speaker
Sponsored by The Ohio State University Discovery Theme
Initiative in Infectious Disease
“Combatting Emerging Viruses: One Health Approach”
As a professor of virology in Rotterdam and Utrecht, The
Netherlands, and in Hannover, Germany, Ab Osterhaus has
a long track record as a scientific researcher and PI of
numerous major scientific projects.
At Erasmus MC, he ran the > 40-person virology diagnostic
lab and the > 100-person virology research lab. His research
programme followed a novel integrated “viroscience” concept, bringing together world-leading
scientists in molecular virology, immunology, epidemiology, pathogenesis, and intervention
studies on human and animal virus infections.
After having handed over the chairmanship of the Erasmus MC Viroscience lab in 2014, he is
currently establishing new One Health Institutes in Utrecht and Hannover. Major performances
include the discovery of more than 50 new viruses of humans and animals (e.g. human
metapneumovirus, human coronaviruses, influenza viruses), elucidation of the pathogenesis of
major human and animal virus infections, and development of novel intervention strategies. This
has enabled health authorities like WHO to effectively combat disease outbreaks like SARS and
avian influenza. The spin-off, Viroclinics Biosciences BV, is another societally relevant success,
allowing effective testing and refining of diagnostic tools and other intervention strategies.
Awards, prizes, guest lecture invitations, co-organizer-ship of international meetings and
editorships of scientific journals further highlight his international recognition. He has acted as
PhD mentor for more than 75 students, holds several key patents and is the author of more than
1100 papers in peer-reviewed journals, together cited more than 50,000 times, and his current
H index is 97. He is editor-in-chief of two major Elsevier Journals. Most of all, Ab Osterhaus
firmly believes scientists have a role to play in translating their knowledge for the benefit and
protection of society.
17
AVMA/AVMF Young Investigators Award
Finalists
Moderated by
Harm HogenEsch, DVM, PhD
Professor, Department of Comparative Pathobiology
Purdue University
Harm HogenEsch is Professor in the Department of
Comparative Pathobiology and Associate Dean for
Research and Graduate Programs in the College of
Veterinary Medicine. He grew up in the Netherlands and
earned his DVM degree cum laude from the University of
Utrecht in 1984. He studied in a combined pathology training/PhD program at the University of
Illinois at Urbana-Champaign from 1984 to 1989, where he did his graduate research in
Immunology under the mentorship of Dr. Peter Felsburg. He became a board-certified
veterinary pathologist in 1989. He then worked as a staff scientist at the TNO Institute for Aging
and Vascular Research in Leiden, the Netherlands from 1989 to 1993. He joined Purdue
University in 1993 and was promoted to Professor in 2001. His research interests include
vaccine adjuvants, vaccine formulation, and the pathogenesis of chronic inflammatory diseases.
He has published over 130 peer-reviewed journal articles and 18 book chapters. His research
has been funded by the NIH, USDA and various foundations and companies.
AVMA/AVMF FIVE YOUNG INVESTIGATOR AWARD FINALISTS:
1. Elshafa Ahmed, The Ohio State University
2. Yung-Yi Mosely, Purdue University
3. Megan Niederwerder, Kansas State University
4. Maciej Parys, Michigan State University
5. Nora Springer, Cornell University
See page 86 for abstracts from the Young Investigator Award finalists.
18
Cheryl London, DVM, PhD
Research Professor
Tufts School of Veterinary Medicine and Molecular Research
Institute at the Tufts Medical Center
Associated Faculty Professor
Feature Speaker
Sponsored by The Ohio State University Comprehensive
Cancer Center and College of Medicine
“Leveraging Comparative Oncology to Maximize
Translational Outcomes”
Dr. London is a Research Professor at Tufts School of Veterinary Medicine and Molecular
Research Institute at the Tufts Medical Center and an Associated Faculty Professor at The Ohio
State University College of Veterinary Medicine (OSU CVM). She is Director of the Clinical
Trials Office at the OSU CVM and Director of Translational Therapeutics at the Center for
Clinical and Translational Sciences, OSU College of Medicine. Prior to her time at OSU, she
was an Assistant Professor in the Department of Surgical and Radiological Sciences at the
University of California Davis. Dr. London earned her DVM at Tufts University and completed
her residency in medical oncology at the University of Wisconsin-Madison and her PhD in
Immunology at Harvard University, where she was also a postdoctoral fellow in the Department
of Pathology. Her research interests center primarily on targeted therapeutics and
translational/comparative oncology.
19
BREAKOUT SESSION A – RESEARCH CAREER PATHWAYS: THREE JOURNEYS
Sponsored by the Society of Toxicologic Pathlogy
Krista M. D. La Perle, DVM, PhD, DACVP
Associate Professor, Veterinary Biosciences
Director, Comparative Pathology and Mouse Phenotyping
Shared Resource
Director, Combined Pathology Residency/Graduate
Program
“Comparative Pathology of Animal Models,
Collaborative Research and Core Facility
Development: One Pathologist’s Research Hat Trick!”
Dr. La Perle joined the faculty in the Department of Veterinary Biosciences at The Ohio State
University College of Veterinary Medicine as Associate Professor in 2008 and serves as
Director of the Comparative Pathology and Mouse Phenotyping Shared Resource. She received
her BS and DVM degrees from North Carolina State University. She then completed
postdoctoral training in veterinary anatomic pathology and obtained her Ph.D. in Experimental
and Molecular Pathology from The Ohio State University where her National Institutes of Healthfunded research focused on animal models of thyroid cancer and developing gene therapy
strategies of sodium/iodide symporter-mediated radionuclide imaging and therapy for cancer.
Prior to returning to OSU, she was Director of the Laboratory of Comparative Pathology and
Genetically Engineered Mouse Phenotyping Service for Memorial Sloan-Kettering Cancer
Center, Weill Medical College of Cornell University and Rockefeller University in New York. Her
primary responsibilities involve providing clinical and anatomic veterinary pathology support to
investigative staff utilizing animal models to study human and veterinary disease, with an
emphasis on conducting extensive baseline phenotypic profiles of genetically engineered
animals and preclinical efficacy and toxicity studies. Dr. La Perle is actively engaged in
mentoring of pathology and laboratory animal trainees, serves as Director of OSU’s Combined
Pathology Residency/Graduate Program, and delivers continuing education for the Charles
Louis Davis DVM Foundation and the Charles River Short Course. She is also very involved in
the American College of Veterinary Pathologists, including 5 previous years on the Examination
Committee, prior Chair of the Student Chapter Committee, previous member of the Certifying
Examination Board, and a senior Councilor. Her publication record includes over 72
manuscripts in peer-reviewed journals and 6 book chapters, as well as numerous abstracts and
presentations at international meetings. When she isn’t working, you can find her scuba diving,
riding her purple Heritage Softail Classic Harley, or watching Columbus Blue Jackets hockey
games!
20
Sarah H. Tannehill-Gregg, DVM, PhD, DACVP
Director, Toxicology
Takeda Pharmaceuticals International Company
“A Career in Industry as a Toxicologic Pathologist:
My Journey in Joining the “Dark Side”; Twists,
Turns, Forks in the Road and Loving What I Do!”
Dr. Tannehill-Gregg received her BA in Zoology from
Miami University (Oxford, OH; 1992), DVM from The
Ohio State University (1996), board certification
(anatomic pathology) from the American College of
Veterinary Pathologists (ACVP, 2004), and PhD from The Ohio State University, Department of
Veterinary Biosciences (2005).
She started out her career as a private practice small animal associate veterinarian directly out
of veterinary school in 1996. Quickly realizing that her love for pathology, which started with a
fascination with dead animals as a child, was piqued with an awesome histology course in
undergraduate training and fine-tuned throughout veterinary school was more than a passing
fancy, she left private practice for graduate school and an anatomic pathology residency in
1997. Under the mentorship and in the laboratory of Dr. Thomas Rosol, Sarah investigated the
pathogenesis of humoral hypercalcemia of malignancy (including the roles of transforming
growth factor β and the Smad signaling proteins), the pathogenesis of skeletal metastases in
breast and lung cancer, and developing animal models of cancer and bone metastasis. In 2000,
she was awarded a prestigious NIH/NCI K08 Mentored Clinical Scientist Development Award to
fund her work.
In 2005, Sarah started her career as a toxicologic pathologist in industry at Bristol-Myers Squibb
in Mt. Vernon, Indiana, where some of her activities included reading and peer-reviewing
toxicology studies and acting as the toxicology representative on multi-disciplinary drug
development teams, where she participated in communicating risk for the use of new drugs in
humans to global regulatory agencies. In 2011, Sarah left Bristol-Myers Squibb to work as a
toxicologic pathologist for Amgen in Thousand Oaks, California, where she continued her work
in pathology and toxicology, assessing and communicating risk, and where she was able to add
experience in protein-based (large molecule) therapeutics. As of March, 2016, she is the
Director of Toxicology at the Boston site of Takeda Pharmaceuticals International Company,
where she looks forward to continuing her work in bringing safe, effective drugs to patients in
need.
21
Joelle Fenger, DVM, PhD, DACVIM (Oncology)
Assistant Professor, Veterinary Clinical Sciences
“Of Mice, Dogs and Men: A Career in Comparative and
Translational Oncology”
Dr. Joelle Fenger received her D.V.M. from the University of
California-Davis School of Veterinary Medicine in 2007 and
completed a Small Animal Medicine and Surgery rotating
internship at The Ohio State University Veterinary Medical
Center in 2008. She entered a combined PhD/Medical
Oncology Residency in the OSU Comparative and Veterinary
Medicine Graduate Program, and in 2013, she received her
board certification in oncology by the American College of Veterinary Internal Medicine. Dr.
Fenger completed her PhD in the OSU Comparative and Veterinary Medicine Graduate Training
Program and joined OSU CVM Department of Veterinary Clinical Sciences as a ResearchIntensive Assistant Professor in May 2015.
As a veterinary oncologist, Dr. Fenger’s work using spontaneous models of cancer in dogs has
focused on the comparative biology of canine and human cancers with the goal of identifying
common molecular pathways that provide a foundation for translational clinical trials. Dr.
Fenger’s research focuses on the role of microRNA dysregulation in spontaneous canine
malignancies. Her laboratory previously identified miR-9 as a mediator of invasive behavior and
metastasis in canine mast cell tumors and osteosarcoma. Dr. Fenger was awarded an NIH K01
training grant to study the role of miR-9 in mast cell biology in vitro and in a transgenic miR-9
mouse model she has developed with collaborators at the OSUCCC-James. She has ongoing
collaborations with researchers at the OSUCCC-James and Nationwide Children’s Hospital
investigating the molecular and biological consequences of miR-9 in osteosarcoma and, more
broadly, studying the impact of microRNA dysregulation in canine and human sarcomas.
22
BREAKOUT SESSION B – TRANSLATIONAL ONCOLOGY
Sponsored by The Ohio State University’s Comprehensive Cancer Center/College of Medicine
Matthew Breen, PhD, CBiol, FRSB
Oscar J. Fletcher Distinguished Professor of Comparative
Oncology Genetics
North Carolina State University
“Comparative Oncology: What Can Dogs (and Sea Lions)
Teach Us About Cancer?”
Matthew Breen completed his PhD in cytogenetics in 1990 and
then worked as a postdoc at the UK Medical Research Council’s
Human Genetics Unit in Edinburgh developing new techniques
as part of the emerging human genome project. After four years
at the University of Queensland, Australia, he returned to the UK
where he developed molecular cytogenetics reagents, resources and techniques for canine
genome mapping and cancer studies. In 2002 Dr. Breen relocated his laboratory to NCSU’s
College of Veterinary Medicine. Over the past decade, Dr. Breen and his team have developed
a series of cytogenetic tools and applied these to investigate cytogenetic changes in numerous
canine cancers. These studies have resulted in the development of molecular assays to
enhance diagnosis, early detection and prognosis. Taking a comparative approach, these
studies are also advancing what we know about human cancers and helping accelerate cancer
gene discovery. Dr. Breen’s research interests also include genome mapping, comparative
genomics, the genomics of military working dogs, and veterinary/wildlife forensics.
Dr. Breen is a Professor of Genomics in the Dept. of Molecular Biomedical Sciences at the
NCSU College of Veterinary Medicine and Associate Director of the NC State Forensic
Sciences Institute. He is a member of the NCSU Comparative Medicine Institute (CMI),
the Center for Human Health and the Environment (CHHE). Dr. Breen is on the steering
committee of the Consortium for Caine Comparative Oncology (C3O) in partnership with the
Duke Cancer Institute, a member of the the Cancer Genetics Program at the University of North
Carolina's Lineberger Comprehensive Cancer Center and a visiting scientist with the
Broad Institute of MIT and Harvard. Dr. Breen was a charter member, and serves on the Board
of Directors, of the Canine Comparative Oncology and Genomics Consortium (CCOGC), a
501c3 not-for-profit organization established to promote the role of the dog in comparative
biomedical research. He is also a charter member of the Sea Lion Cancer Consortium (SLiCC).
23
Ryan Roberts, MD, PhD
Physician-Scientist
Nationwide Children’s Hospital
Columbus, Ohio
“Enlisting Man’s Best Friend in the Fight Against His
Children’s Worst Enemy: Seeking Common Cures”
Dr. Roberts works as a physician-scientist at Nationwide
Children’s Hospital in Columbus, Ohio. His laboratory
focuses on identifying mechanisms of metastasis in
pediatric solid tumors—discovering how tumors spread to and grow in specific organs. His work
in osteosarcoma has demonstrated that IL-6 and IL-8 play critical roles in the tumor-lung
interactions that facilitate lung metastasis. His current work explores whether targeting these
pathways might prove useful in preventing or even treating metastatic disease. Dr. Roberts has
developed close relationships with his veterinary colleagues and sees great potential in
accelerating the development of novel therapeutics through coordinated investigation in both
client animals and children suffering from osteosarcoma.
24
FRIDAY EVENING
SPEAKERS
JULY 29, 2016
25
Caroline C. Whitacre, PhD
Senior Vice President for Research
Caroline C. Whitacre, Ph.D. serves as the Senior Vice President for
Research at The Ohio State University in Columbus, Ohio. She is a
Professor of Microbial Infection & Immunity and her research area is the
immunology of multiple sclerosis. She served as Associate Vice
President for Health Sciences Research and Vice Dean for Research in
the College of Medicine from 2002 to 2008. Prior to that, she served for
12 years as the Chair of the Department of Molecular Virology,
Immunology and Medical Genetics in the College of Medicine. In her
current role as VP for Research, Dr. Whitacre is responsible for the overall strategic planning
and infrastructure support for the university’s $983 million annual research program.
In recognition of her university activities, she was awarded the OSU Faculty Award for
Distinguished University Service in 2001 and the Distinguished Scholar Award in 2008. In 2004,
she was named a Fellow of the American Association for the Advancement of Science. In
2015, she was inducted into the National Academy of Inventors.
She serves on the boards of Rev1 Ventures of Columbus, Center for Science and Industry
(COSI), The Transportation Research Center, The Wellington School, BioOhio, the Multiple
Sclerosis Society and Oak Ridge Associated Universities. She chairs the Board of SciTech, the
Ohio State University Research Park.
Dr. Whitacre received her BA and PhD degrees at The Ohio State University.
Brad Fenwick, DVM, PhD
Senior Vice President Global Strategic Alliances
Elsevier
Dr. Brad Fenwick is a Professor of Pathobiology and Microbiology.
He holds a doctor of Veterinary Medicine and Masters of Pathology
from Kansas State University and Ph.D. in Comparative Pathology
from UC Davis. He completed a residency at UC Davis and is
board certified by the American College of Veterinary Microbiology
and board qualified in Pathology. Dr. Fenwick has received
numerous awards and recognitions for his research, holds several
patents, founded a biotechnology company, and consults globally
with companies, universities, and governments. He is a Fellow with
the American Council on Education, a Fellow with the American
Association for the Advancement of Science, and is currently a Jefferson Science Fellow and
Senior Science Advisor with the U.S. State Department where his portfolio includes Food
Security, Higher Education, and Science, Technology, and Innovation. He has held many
senior administrative positions, including Graduate Dean, Vice President for Research,
President for Intellectual Properties, Vice Chancellor for Research and Engagement, and Chief
Scientist with the USDA. As Senior Vice President for Global Strategic Alliances with Elsevier
he is charged with forging non-commercial partnerships with universities, research centers,
industry, governments, and funding bodies to enhance higher education and academic research
and innovation productivity and success.
26
Jan Ramer, DVM, DACZM
Director of Conservation Medicine/Staff Veterinarian
The Wilds
Feature Speaker
Sponsored by Elsevier
“One Health/Conservation Medicine, Abroad and in
Your Backyard”
Dr. Jan Ramer joined the Wilds as Director of
Conservation Medicine/Staff Veterinarian in February
2015. Jan’s route to veterinary medicine was somewhat
non-traditional. She earned her B.S. in Biology from Purdue University and used that education
in a 12-year career as an animal keeper, first at the Indianapolis Zoo followed by 10 years at the
Brookfield Zoo. During that time, she participated in lemur behavioral studies in Madagascar
and traveled to Rwanda in 1985 where she was fortunate to meet Dian Fossey and the majestic
mountain gorillas.
While she loved her career as an animal keeper, Jan was drawn to veterinary medicine, so she
made a career change, receiving her DVM from the University of Wisconsin in 1995. With the
goal of performing conservation medicine, she first worked in private practice and taught at the
University of Wisconsin. She then moved to the Indianapolis Zoo as the Associate Veterinarian
for 14 years. Jan became a Diplomate of the American College of Zoological Medicine in 2007.
She also served as President of the American Association of Zoological Veterinarians. She was
a member of the International Iguana Foundation board of directors and of the IUCN Iguana
Specialist group for many years. Jan found her passion in conservation medicine as Regional
Manager for the Gorilla Doctors, where she worked with a wonderful team of African
veterinarians in Rwanda, Uganda and eastern Congo for 3 ½ years, returning to the USA in
January 2015.
In her leisure time, Jan loves spending time with her family and friends. She is thrilled to be a new
grandma and tries to spend as much time with little Ben as possible!
Jan is excited to join the Wilds. She shares that “the Wilds has fantastic species in a unique
setting, is involved in great conservation science, and the dedicated staff feels like a family – it is an
honor to be part of the team!”
27
SATURDAY SPEAKERS
JULY 30, 2016
28
James (Jimi) Cook, DVM, PhD, OTSC
William and Kathryn Allen Distinguished Professor in
Orthopaedic Surgery
Director, Comparative Orthopaedic Laboratory,
Orthopaedic Research Division and Mizzou BioJoint
Center
University of Missouri
Feature Speaker
Sponsored by the Ohio Veterinary Medical Association
“One Health – One Medicine: How Veterinary Medicine
is Fixing People Too”
After receiving his B.S. degree from Florida State
University and competing for 5 years as a professional waterskier, Dr. James (Jimi) Cook
completed his DVM in 1994 and PhD in 1998. In 1999, he founded the Comparative
Orthopaedic Laboratory at the University of Missouri, a multi-disciplinary team of physicians,
veterinarians, engineers, and basic scientists dedicated to translational orthopaedic research.
He has over 180 peer-reviewed publications and over $20 million in research funding, has
received numerous awards including America’s Best Veterinarian (2007), holds 14 US patents
and has seen 3 biomedical devices through FDA approval to human clinical trials. He is
currently Director of the Mizzou BioJoint Center, Director of The Comparative Orthopaedic
Laboratory and the William and Kathryn Allen Distinguished Professor in Orthopaedic Surgery,
and serves as Director of the Division of Research for the Department of Orthopaedics at the
University Hospital’s Missouri Orthopaedic Institute. He is also co-founder of Be The Change
Volunteers a NGO dedicated to building schools in remote villages in the developing world
whose teams have built 32 educational facilities in 15 countries, providing educational
opportunities to more than 6,000 students.
MERIAL VETERINARY RESEARCH SCHOLAR AWARD
Presented by Monica D. Figueiredo, DVM, PhD, DACVIM,
Merial, Director Lead Finding and External Innovation
Laura LoBuglio, third year veterinary student at the University of Minnesota
“Identification and Inhibition of Oxidative Stress-induced Apoptosis Pathways in Beta
Cells in Diabetes”
MERIAL VETERINARY RESEARCH GRADUATE STUDENT AWARD
Presented by Diane Larsen, DVM, PhD, Merial Head, Pharma Development Projects
Victoria Baxter, Postdoctoral Fellow, Johns Hopkins Bloomberg School of Public
Health
“The Role of Interferon Gamma in the Immunopathogenesis and Control of Sindbis Virus
During Nonfatal Alphavirus Encephalomyelitis”
29
AVMF/WINN FELINE FOUNDATION RESEARCH
AWARDEE
Andrea J. Fascetti, VMD, PhD, DACVN, DACVIM
Scientific Director, Amino Acid Laboratory
Scientific Director, Feline Nutrition and Pet Care Center
Scientific Director, Feline Research Laboratory
Andrea Fascetti graduated from the University of
Pennsylvania School of Veterinary Medicine. Following graduation she completed an internship
and medicine residency at The Animal Medical Center in New York City. She holds a doctoral
degree in nutrition from the University of California, Davis. She is a diplomate of both the
American College of Veterinary Internal Medicine and the American College of Veterinary
Nutrition. Andrea is currently a Professor of Nutrition at the University of California, Davis. She
is the service chief for the Nutrition Support Service in the Veterinary Medical Teaching
Hospital, as well as the Scientific Director of the Feline Nutrition and Pet Care Center, the Feline
Research Laboratory and the Amino Acid Laboratory.
She has authored over 60 peer-reviewed publications. She also co-edited a textbook entitled,
“Applied Veterinary Clinical Nutrition.” Andrea has served as the Secretary/Treasurer of the
American College of Veterinary Nutrition and as a member at large on the executive board.
Her current research interests are trace mineral and amino acid metabolism, obesity, carnivore
nutrition, nutritional idiosyncrasies of the cat, improvement of pet foods and clinical nutrition.
30
AVMA LIFETIME EXCELLENCE IN RESEARCH
AWARDEE
Susan M. Stover, DVM, PhD, DACVS
Professor, Department of Anatomy, Physiology & Cell
Biology
Director, JD Wheat Veterinary Orthopedic Research
Laboratory
University of California, Davis
Dr. Stover is a Professor and Director of the JD Wheat
Veterinary Orthopedic Research Laboratory at the University
of California at Davis. She received her veterinary degree from Washington State University
and subsequently completed an Equine Surgery Internship and Residency at the University of
California at Davis. She was in equine practice in Washington State before returning to the
Veterinary Medical Teaching Hospital, UC Davis to teach clinical equine lameness and surgery
to veterinary students and residents. She became board certified by the American College of
Veterinary Surgeons while pursuing a PhD program focused on equine orthopedic research
(dorsal metacarpal disease (‘bucked shins’) in Thoroughbred racehorses). She now devotes
her time to equine orthopedic research, with over 200 research publications; mentoring
veterinary students, graduate students, and residents in research; and teaching musculoskeletal
anatomy, biomechanics, and pathology to veterinary students.
Her major research focuses are the prevention of musculoskeletal injuries in equine athletes
and the biomechanical function of musculoskeletal structures and treatment of orthopedic
disorders in domestic and non-domestic animals. Her key contributions to the safety and welfare
of the horse include development of arthrocentesis (joint fluid sampling) techniques, discovery
of lesions that predispose to catastrophic injuries in racehorses, and elucidation of factors that
contribute to injury development in racehorses. Current research efforts are focused on
understanding how training and injury affect bone adaptation or propensity for bone fracture, the
effects of performance surface materials and shoes on hoof and fetlock biomechanics and thus
propensity for injury in athletes, and the development of osteoporosis in horses associated with
inhalation of cytotoxic silicate-laden soil.
31
BREAKOUT SESSION C – REGENERATIVE MEDICINE
Sponsored by the Ohio Veterinary Medical Association
Alicia L. Bertone, DVM, PhD, DACVS, DACVSMR
Professor and Equine Orthopedic Surgeon
Director, Comparative Orthobiologics Research Laboratory
“Hybrid Veterinary Clinical Trials in Regenerative Medicine”
Dr. Bertone received her Bachelor of Science degree (1977),
DVM degree (1982) and internship (1983) from Cornell
University, subsequently completed a combined surgery
residency/PhD program (1987) at Colorado State University and
joined the surgery faculty at Louisiana State University that same
year. Dr. Bertone became board certified as a Diplomate in the
American College of Veterinary Surgeons in 1988 and joined the surgery faculty at The Ohio
State University late in 1989. Dr. Bertone focused her clinical and research efforts in the field of
orthopedics and became a full professor in 1997. Dr. Bertone has served as an equine
orthopedic surgeon at The Ohio State University Galbreath Equine Center for over 25 years,
managing cases and training veterinary students and equine surgery residents. Dr. Bertone has
mentored many PhD and Masters of Science graduate students, surgery residents and research
fellows and has developed a reputation for a quality and productive research program and a
strong orthopedic elective surgical caseload. Dr. Bertone has been recognized as one of the top
extramurally funded researchers in the College of Veterinary Medicine and has greater than 160
peer reviewed scientific publications. After completing a 1-year sabbatical at the Center for
Molecular Orthopedics at Harvard University (2000), Dr. Bertone has served two terms as the
Trueman Family Endowed Chair and established the Comparative Orthopedic Molecular
Medicine Suite and Applied Laboratories that specializes in regenerative medicine including
stem cell and growth factor therapies. Dr. Bertone has retained a clinical appointment at the
Galbreath Equine Center at The Ohio State University where she regularly performs surgery.
Dr. Bertone is a member of the The Sports Health and Performance Institute, The Dorothy M.
Davis Heart and Lung Research Institute, and The Regenerative Medicine and Cell Based
Therapy Center, and is an adjunct full professor in the Department of Orthopedics in the College
of Medicine. Dr. Bertone became board certified in Equine Sports Medicine and Rehabilitation in
2013. Dr. Bertone’s clinical focus is orthopedic surgery and her research focus is in the study of
comparative orthopedic medicine, regenerative medicine, and gene therapy for the treatment of
cartilage injury and bone repair. Dr. Bertone has been active in veterinary and human
orthopedic associations and has been regularly invited to participate in national and
international scientific meetings because of her expertise in regenerative medicine, orthopedics
and cell-based therapies.
32
Kurt D. Hankenson, DVM, PhD
Professor
Small Animal Clinical Sciences
Michigan State University
“Developing Novel Therapies for Bone Regeneration: A
Stem Cell Biology Approach”
Dr. Hankenson received his DVM (veterinary degree) from
the University of Illinois (1992), an MS from Purdue
University (1997) and his PhD from the University of
Washington, Department of Biochemistry (2001). He is currently the Co-Director of the
Laboratory for Comparative Orthopaedic Research (LCOR) at Michigan State University. A
former equine veterinarian, he began his independent research career at the University of
Michigan in 2002 as a faculty member in the Orthopaedic Research Laboratories (ORL). In
2006 he moved to the University of Pennsylvania School of Veterinary Medicine, where he was
the inaugural holder of the Dean W. Richardson Chair for Equine Disease Research. Dr.
Hankenson is an American Society for Bone and Mineral Research (ASBMR) Young
Investigator award winner (2002), received a John Haddad Fellowship from the ASBMR (2003),
and in 2008 was the first veterinarian awarded the Fuller Albright award by the ASBMR.
Dr. Hankenson has published over 85 peer-reviewed manuscripts. He is a former member of
the Scientific Medical Advisory Council for the Canadian Arthritis Network, is a member of the
Grayson Jockey Club Research Advisory Committee, and is a permanent member of both the
Veterans Affairs Medical Council Endocrinology B (EndoB) study section and the NIH Skeletal
Biology Structure and Regeneration (SBSR) study section. He has previously served on the
Board of Directors for both the Orthopaedic Research Society (ORS) and Advances in Mineral
Metabolism (AIMM), and is chair-elect for the North American Veterinary Regenerative Medicine
Association. Dr. Hankenson is an associate editor for Connective Tissue Research and the
Journal of Orthopaedic Research and an editorial board member of the Journal of Experimental
Orthopaedics. He has been the recipient of numerous extramural grants as both PI and coinvestigator (NIH, DoD, and DOE), receiving over $15 million in extramural funding since 2002.
He has submitted two US patents related to therapies to accelerate bone formation and repair.
Programmatically, his laboratory probes mechanisms regulating osteoblast differentiation and
bone formation and regeneration. He is focused on translating novel basic research findings to
clinically applicable therapies to heal and restore lost bone.
33
Daniel Gallego Perez, PhD
Assistant Professor, Surgery and Biomedical Engineering
Director, Nanomedicine Laboratory
“Nanotechnology-Enabled Regenerative Medicine”
Dr. Gallego-Perez is an assistant professor in the departments of
Surgery and Biomedical Engineering at The Ohio State
University. He is the director of the Nanomedicine Lab, which
focuses on the development and implementation of novel microand nanoscale technologies (or devices) for therapeutic or
fundamental applications. His major areas of research include gene delivery/therapy, cellular
reprogramming, lab-on-a-chip platforms, biomaterials, and tissue engineering.
BREAKOUT SESSION D – INFECTIOUS DISEASES
Sponsored by Ohio State’s Discovery Theme Initiative in Infectious Disease
Michael Oglesbee, DVM, PhD, DACVP
Professor and Chair, Department of Veterinary Biosciences
Director, Ohio State CVM Summer Research Program
Faculty Lead, University Discovery Theme in Infectious Disease
“Fever and Virus Infection of the Brain: Good, Bad and Ugly?”
Dr. Oglesbee is professor of virology and veterinary neuropathology
in the Department of Veterinary Biosciences, College of Veterinary
Medicine, The Ohio State University. Scientific contributions have
focused on host determinants of viral neurovirulence, and he was
named a fellow by the American Association for the Advancement of
Science in recognition of distinguished contributions to our
understanding of virus-heat shock protein interactions related to
infection, virulence, and impact on innate and adaptive antiviral immunity. Dr. Oglesbee is chair
of the Department of Veterinary Biosciences, faculty director of the College Summer Research
Program for veterinary medical students at The Ohio State University, and principal investigator
on an NIH training grant (T35) in support of that program. Dr. Oglesbee leads the college’s
signature program in Infectious Disease, and is also the faculty lead on a university investment
in infectious disease involving the colleges of Veterinary Medicine, Medicine, Nursing, Food
Agriculture and Environmental Sciences, Public Health, Pharmacy, and Arts and Sciences, and
the Food Animal Health Research Program at the Ohio State Wooster Campus. Focus of the
investment is development of interdisciplinary collaborative research networks in the
programmatic areas of antimicrobial resistance mechanisms and drug discovery, treatment and
prevention of virus infection, host response to infection and microbial communities, and the
ecology of pathogen emergence and spread. The ultimate goal is to enhance our ability to
control the spread and severity of infectious disease through the creation and dissemination of
knowledge, practices and products.
34
Larry S. Schlesinger, MD
Samuel Saslaw Professor of Medicine
Chair, Department of Microbial Infection and Immunity
Director, Ohio State Center for Microbial Interface Biology
“Approaches to the Study of Tuberculosis”
Larry S. Schlesinger earned a BA in Biology from Cornell University
and MD from Rutgers Medical School. He completed his residency
in Internal Medicine at the University of Michigan and clinical and
research fellowships in Infectious Diseases at UCLA. He joined the
faculty at The University of Iowa in 1991 where he rose from
Assistant Professor to Professor of Medicine and served as
Fellowship Director for the Division of Infectious Diseases and Associate Chair of the
Department of Medicine. He moved to The Ohio State University in 2002 where he served as
Director of the Division of Infectious Diseases, Department of Internal Medicine until 2011. He is
currently the Samuel Saslaw Professor of Medicine, first Chair of the Department of Microbial
Infection and Immunity and founding Director of the OSU Center for Microbial Interface Biology,
an interdisciplinary campus-wide program that focuses on infectious diseases of major concern
to human health. He is a cellular immunologist whose studies focus on the pathogenesis of
tuberculosis and other airborne infections due to intracellular pathogens that subvert lung
immune mechanisms. His discoveries have led to greater insight into the unique attributes that
soluble and cellular components of the innate immune system of humans bring to the microbehost interface, translating them into drug discovery platforms. He is a current NIAID Council
member; fellow of the AAAS, AAM and AAP; and OSU’s 2011 Distinguished Scholar and 2105
COM Distinguished Professor.
Dr. Schlesinger has placed great emphasis on education and mentoring throughout his career,
particularly in clinical and translational research, and has been committed to building strong
interdisciplinary academic programs. He has been/is a faculty member of 10 pre- and postdoctoral training programs (NIH and HHMI) and is PI of 2 NIH T32 training grants, including the
MSTP. In all, he has mentored ~135 trainees at all levels, several of whom have been awarded
national research fellowships (34 in total) and have gone on to academic or industry positions.
He became director of the OSU Medical Scientist Program (MD PhD granting) in 2008 and was
awarded the first ever NIH-funded MSTP in 2011 (renewed in 2016). He is a current member of
the AAMC GREAT MD-PhD Section Steering Committee and chair-elect of this group.
35
Gabriel L. Hamer, PhD
Assistant Professor
Department of Entomology
Texas A&M University
“Evaluating the Relative Role of Different Mosquito Species to
the Transmission of Viruses”
Gabe Hamer earned his Bachelors and Masters degrees at the
University of Illinois in the Department of Natural Resources and
Environmental Science. He obtained his PhD in Fisheries and
Wildlife at Michigan State University and then spent three years as a post-doctoral researcher at
Michigan State University and the University of Wisconsin. Gabe joined the faculty at Texas
A&M University in the Department of Entomology in 2012 and is currently an Assistant
Professor. Gabe’s research broadly investigates the ecology of infectious diseases of humans,
wild animals, and domestic animals, with particular attention to those transmitted by
mosquitoes, ticks, and kissing bugs. Gabe has 38 publications in peer-reviewed journals.
36
Nationwide A
Nationwide B
1st Floor
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North Drop Off
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Abstracts Listed Alphabetically
Abstracts are ordered alphabetically by last name.
Last Name
First Name Poster
#
Abdel Massih
Abeywardena
Adams
Cherein
Shanes
Daniel
1
2
3
Aguiar
Ariel
4
Alexander
Derecka
5
Alexander
Kayla
6
Allgood
Hillary
7
Anderson
Jessica
8
Arica
Andrea
9
Arroyo
Geraline
10
Asklof
Au
Kendra
Gina
11
12
Aust
Avalos-Cavero
Rebecca
Barbara
13
14
Bagnall
Hannah
15
Baker
Anna
16
Baley
Ballard
Bancroft
Susan
Kristen
Channing
17
18
19
Banducci
Caitlin
20
Banuelos
Rosa
21
Barbar
Megan
22
Barnett
Brian
23
Bartholomew
Kristen
24
Abstract Title
Page
#
Procoagulant microvesicles in canine packed red blood cells
Application of endocrine techniques for wildlife conservation
Determining the effects of SMAC mimetic drugs on vascular
endothelial cells
Characterizing immune regulation of canine heartworm Dirofilaria
immitis infection in Aedes aegypti mosquitoes
Socioeconomic status of population and negligence of dogs with
visceral leishmaniasis in Araçatuba-SP, Brazil
Flow cytometric evaluation of canine lymphocytes after oral
mycophenolate mofetil
Surveillance of Florida snails for Parelaphostrongylus andersoni and
Parelaphostrongylus tenuis
The impact of a cancer-specific diet on inflammation and side effects
associated with chemotherapy in dogs
Investigating the effects of EZH2 inhibition in canine B-cell
lymphoma
Comparison of MALDI-TOF and PFGE for strain-typing
Staphylococcus aureus isolated from cow’s milk
Determining the role of RIPK3 in early responses to Yersinia infection
Combinatory radiotherapy and cellular immunotherapy enhances
tumor cell killing
The role of CDK9 protein in transcription of herpes simplex virus I
Study of human and mouse lung cells responses to carbon nanotubes
to assess their pulmonary disease potential
An investigation into the epidemiology of B. odocoilei in Ontario ticks
and cervids
Volumetric analysis of the hippocampus and amygdala in California
sea ions with domoic acid induced epilepsy
Evaluation of a point-of-care coagulometer in two raptor species
Detecting circulating DNA in dogs with mast cell tumors
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91
92
Effects of 17-a-estradiol treatment on stifle joint morphology in aging
mice
Impact of Paternal and Maternal Preconception Stress on Offspring
Neurodevelopment and Stress Response
Prevalence and pathology of Trypanosoma cruzi in coyotes and
raccoons in a disease-endemic region of Texas
Evaluation of improvement in joint function from intra-articular
injection of NV-08 in dogs with OA joint pain
Assessing key proinflammatory cytokines in equine peripheral blood
mononuclear and monocytic cells, in vitro
Novel in vitro model system of canine degenerative myelopathy using
direct conversion of skin fibroblasts
40
92
93
93
94
94
95
95
96
96
97
97
98
98
99
99
100
100
101
101
102
102
Last Name
First Name Poster
#
Baumgardner
Rachel
25
Beasley
Erin
26
Becher
Jessica
27
Belling
Belter
Carolyn
Rebecca
28
29
Bennett
Joshua
30
Bentz
Kelsey
31
Bickers
Bridget
32
Binns
Jamie
33
Binstock
Jonah
34
Birkner
Elise
35
Bishop
Bishop
Brianne
Kaitlin
36
37
Black
Kelley
38
Blank
Carolyn
39
Bloom
Boire
Christopher
Francois
40
41
Bollman
Mary Kate
42
Brado
Glenn
43
Briggs
Brockhurst
Cassidy
Jacqueline
44
45
Bucak
Burns
Emily
Jordan
46
47
Byerley
Cabral
Sydney
Troy
48
49
Caceres
Cahill
Valeria
Bridgette
50
51
Abstract Title
Page
#
Investigating IgG subtype responses in horses infected with different
mutations of EHV-1
Pet carriage of Staphylococcus aureus and S. pseudintermedius in the
households of children with asthma
Hormonal effects on adenoma development in a rat model of human
familial colon cancer
Developing selective growth media for Ophidiomyces ophiodiicola
Comparison of enterotomy leak pressure among fresh, cooled, and
frozen/thawed swine jejunal segments
Acute toxicity evaluation in rats of potential chemical warfare agent
antidotes
Susceptibility of Amblyomma americanum, Dermacentor variabilis
and Rhipicephalus sanguineus to acaricides
The Effect of High Density Lipoproteins on Bovine Neutrophil
Activation
Sympathetic neurotransmitter induced vasoconstrictions in high fat
and control diet fed rats
Col3’s roles in the fibrotic response: therapeutic strategies to engineer
regenerative and tumor environments
Trypanosoma cruzi strain types infecting triatimone vectors and
primates at two research facilities in Texas
Characterizing changes in bovine veterinary medicine in Missouri
Pharmacodynamic assessment of a panel of immunosuppressant drugs
on ex-vivo canine T-lymphocyte proliferation
Development and performance analysis of a multiplex assay to detect
exposure to tick-borne diseases in dogs
Variation in bronchial size during respiration and cough in healthy
compared to malacic dogs
Characterizing apoptosis in a zebrafish mdm1 mutant
Optimizing the broth environment for large volume growth of
“Brachyspira hampsonii”
Application of additive manufacturing technology and stem cell
recruitment to improve osseointegration
Effect of intranasal cannabinoid administration on an experimentally
induced rhinosinusitis mouse model
Effect of resistant starch prebiotic on enteric immune status in swine
Evaluation of Plasmodium knowlesi in vitro susceptibility to
methotrexate
Sperm characteristics associated with bull fertility
Identification of biomarkers for xenoestrogenic exposure using a
prepubertal porcine model
Lacunar-canalicular morphology in tetrapod long bones
Improving adjunctive treatment of drug resistant diabetic foot
infections
Irx3 and Irx5 Disrupt Follicle Integrity in the Ovary
Linking canine distemper virus susceptibility to SLAM sequences in
wildlife species
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104
104
105
105
106
106
107
107
108
108
109
109
110
110
111
111
112
112
113
113
114
114
115
115
116
Last Name
First Name Poster
#
Callahan
Robert
52
Calle
Jasmine
53
Cardwell
Jason
54
Carlson
Ariel
55
Carothers
Currie
56
Carter
Samantha
57
Cash
Kaycee
58
Cashin
Kaitlyn
59
Casper
Castell
Caudill
Daniele
Natalie
Mitchell
60
61
62
Chan
Dennis
63
Chan
Jennifer
64
Chandler
A’Jah
65
Chang
Kevin
66
Charles
Jenna
67
Chen
Aolei
68
Chu
Colin
69
Chung
Sarah
70
Clark
Megan
71
Clark
William
72
Clarke
Samantha
73
Cleveland
Ashley
74
Cluett
Matthew
75
Coleman
Denver
76
Abstract Title
Page
#
Generation of a recombinant PIV5 virus expressing Zika antigen for
vaccine development
The role of Hus1 impairment in non-small cell lung cancer
tumorigenesis
Plasmin induces up regulation of matrix metalloproteinase-13 in
murine bone marrow derived macrophages
The effects of metformin and resveratrol on canine hemangiosarcoma
cell lines
Respiratory responses to short term exposure to second hand smoke in
a Guinea pig asthma model
Biomechanical risk factors in the development of osteochondrosis in
Standardbred pacers and trotters
In vitro evaluation of cell mediated immune responses to respiratory
antigens in high and low risk beef calves
Effect of low starch diet on health of sloth bears (Melursus ursinus) at
Cleveland Metroparks Zoo
Using nature’s trick to effectively deliver DNA for gene therapy
Characterization of Shark Ig/TCR Lymphocytes
The proline utilization system of Brucella abortus is required for
virulent infection
Development and validation of a PCR platform for the sensitive and
specific detection of Bartonella rochalimae
Fluoroscopic determination of thoracic dimensional changes in
healthy dogs
Characterizing the mechanisms of action of novel chemical inhibitors
of Mycobacterium smegmatis growth
Fecal toxin cytotoxicity as a measure of clinical disease in mice
infected with Clostridium difficile R20291
Comparison of NIBP measurements obtained in a veterinary clinic
versus a home setting in hypertensive dogs
Systematic literature review and elicitation of expert opinion to assess
the 1997 FMD epidemic in Taiwan
In vitro endothelial cell wound healing in response to red wine
polyphenols resveratrol and quercetin
Effect of FABP1/SCP-2/SCP-X Gene Ablation (TKO) on the
Endocannabinoid System in Mice
Characterization and optimization of ex vivo T cell expansion for redirected T cell therapy
Early effects of ivermectin on intraperitoneal Brugia pahangi
infection in Meriones unguiculatus
Alterations in myofilament-associated signaling complexes in dogs
with naturally occurring DCM
Anatomy of the lower respiratory system of the African Grey Parrot
(Psittacus erithacus erithacus)
Effects of IL-10 modulation on nerve pathology in C. jejuni induced
Guillain-Barre mouse models
Effect of temperature on Veronaea botryosa growth and cytotoxicity
to Acipenser transmontanus skin cell lines
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118
118
119
119
120
120
121
121
122
122
123
123
124
124
125
125
126
126
127
127
128
128
Last Name
First Name Poster
#
Condrey
Jillian
77
Courville
Cox
Emily
Amanda
78
79
Crilly
Nathan
80
Crombie
Kathryn
81
Croughan
Carolyn
82
Cutcliffe
Ross
83
Daney
Jolani
84
Daniels
Daugherty
De La
Espriella
Dellinger
Barbara
Elizabeth
Jose
85
86
87
Marion
88
Demars
Fanny
89
Devereaux
Dickson
Joseph
Rachel
90
91
Dionne
Madeleine
92
DiPastina
Ann
93
Draper
Janna
94
Duliepre
Dunfield
Eagalle
Stephie-Anne
Elizabeth
Thisuri
95
96
97
Ebelle
Danielle
98
Ehrlich
Allison
99
Emmitt
Nicole
100
Engel
Danielle
101
Esposito
Gena
102
Falsey
Rachel
103
Abstract Title
Page
#
Effect of mesenchymal stem cell conditioned media on TGF-b1
exposed equine fibroblasts
Ectopic Neuronal Connections in a Mouse Model of Autism
Role of inhibitor of differentiation (Id) proteins in fibrosis of the
human cornea
TAM Receptor Expression Is Altered During Zika Virus Infection in
Pregnant Mice
The genetics of familial hypertrophic cardiomyopathy in the Siberian
cat
Assessing mental health and identifying stressors among LSU School
of Veterinary Medicine students and alumni
Cancer Stem Cell-Targeted Vaccine Induces Antibodies Against
Conserved Vaccine Antigens
The prophalactic effect of acetylsalicylic acid on astrogliosis in a
mouse model of scrapie
Investigation of blood lead levels of dogs living in Flint, Michigan
Development of a PCR panel for Tick-Borne Pathogens
Purification and stability screens of conserved virulence factor A in
Streptococcus pyogenes
Do male sticklebacks use visual or olfactory cues to assess the history
of a potential mate?
Does in utero exposure to domoic acid, found in seafood, cause
epilepsy?
At what age does canine gait mature?
Evaluating the effect of Fortiflora probiotic therapy on feline
Tritrichomonas foetus infection
Evaluating the role of female sex hormones on left ventricular
mechanics in aortic-banded mini-swine
Optimizing antimicrobial use for mastitis to combat antimicrobial
resistance on US dairy farms
Microneutralization assay: an economical method for determining
vaccine recommendations for swine influenza
Roles for the 9-1-1 DNA damage response complex in meiosis
A survey of stereotactic radiotherapy in veterinary medicine
Determination of causes of mortality in captive psittacines submitted
to the Ontario Veterinary College
The Effect of Sandstorm Particles on Antigen-Presenting Dendritic
Cells
Breast cancer cell growth mediated by EGFR/HER1 expression is
regulated by p53 suppressor function
Role of Neural Inflammatory Processes in Estradiol Enhancement of
Cocaine Self Administration in Female Rats
Make food great again: effect of protein source and amount on wholebody metabolism and weight gain
Recreating geophagy in a captive psittacine species, Myiopsitta
monachus
Glutathione S-transferase-mu polymorphisms and cancer risk in
boxers
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130
131
131
132
132
133
133
134
134
135
135
136
136
137
137
138
138
139
139
140
140
141
141
142
Last Name
First Name Poster
#
Fasig
Joseph
104
Fedderly
Ferrero
Galya
Nicole
105
106
Fisk
Elizabeth
107
Fleming
Payge
108
Folkerts
Stephanie
109
Ford
Jordan
110
Fousse
Samantha
111
Frady
Kristine
112
Furst
Emmanuelle
113
Furst
Nicole
114
Galang
Kristopher
115
Gavitt
Ashley
116
Gavron
Nancy
117
Geisler
Jennifer
118
Ghearing
Gilfeather
Girens
Natasha
Christa
Renee
119
120
121
Glick
Melissa
122
Guan
Chiyu
123
Gustafson
Kevin
124
Haffner
125
Hagstrom
Abigail
(Abbie)
Melena
Haiderer
Elizabeth
127
Haight
Kimberly
128
126
Abstract Title
Page
#
Evaluating body condition index with DEXA and deuterium oxide in
big brown bats (Eptesicus fuscus)
Pathogenesis of influenza in a murine model of asthma
Development of a qPCR assay for Altered Schaedler Flora colonized
mice utilizing the groEL gene
Comparison of Contemporary Aqueous and Alcohol-Based Scrub
Agents for Rodent Surgical Skin Preparation
Nexelom cellometer live cell and viability verification and error
evaluation for the canine stem cell assay
Isolation of antibiotic-producing bacteria from New River Valley
(NRV) soil
Parvovirus detection by PCR and ISH is associated with myocarditis
and cardiomyopathy in young dogs
Does re-emerging St. Louis encephalitis virus in California show
increased fitness compared to older strains?
Characterization of the pathogenic potential among different species
in the spotted rickettsial group
Primary neuron culture autapses as a model for investigating opioid
receptor desensitization
Cardiovascular effects of epinephrine on Alligator mississippiensis
recovering from isoflurane anesthesia
Marble Burying for Assessing Postoperative Pain in Rats Treated with
Meloxicam or Sustained-Release Meloxicam
Custom 3D-printed drill guides for surgical stabilization of the canine
cervical spine - a cadaveric study
Refining approaches to assess microbiota and low-profile pathogens in
the tick vector, Amblyomma maculatum
miR146a is an endogenous regulator of both hematopoiesis and bone
mass
Preventing infectious diseases in dogs at shows
Cannabidiol Use and Its Potential Immune Effects in Modulating EAE
Calculation of body surface area using computed tomography-guided
modeling in dogs and cats
Estrogen protects progestin-treated mice against intravaginal
transmission of cell-associated HIV-1
Expression and Purification of PrM and E Proteins of Zika Virus in
Sf9 Cells
The use of a B cell tetramer to evaluate the memory immune response
to PRRSv
Investigating novel small compounds and their effect on inflammation
and chondrogenesis in joints
Vertical non-transovarial transmission of Rickettsia felis in the cat flea,
Ctenocephalides felis
Characterization of the spectrum of activity and mechanism of a pHselective inhibitor of M. tuberculosis
A porcine model of vascular cognitive impairment for dementia
prevention
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145
145
146
146
147
147
148
148
149
149
150
150
151
151
152
152
153
153
154
154
Last Name
First Name Poster
#
Hammes
Joseph
129
Hanes
Steven
130
Hanzel
Amanda
131
Hardy
Hargis
Jennifer
Jessie
132
133
Harris
Elizabeth
134
Hausmann
Katharine
135
Hayes
Allison
136
Hayter
Sydney
137
Healey
Eleni
138
Hecker
Kari
139
Hendrikx
Hendrix
Lysanne
Madeleine
140
141
Henson
Christopher
142
Herrold
Emily
143
Holdridge
Julie
144
Horng
Katti
145
Hosner
James
146
Howard
Hristova
Hsu
Jennifer
Teodora
Steven
147
148
149
Huff
Jaimie
150
Hughes
Courtney
151
Hughes
Peyton
152
Hunt
Hunter
Icaza
Janna
Jacquelyn
Melissa
153
154
155
Abstract Title
Page
#
Reproductive performance applied to genetic merit and increasing
insemination rate of anovular Holsteins
Compromising X-linked gene silencing in mature B cells through
deletion of the long noncoding RNA Xist
Effect of a Western-style or a high fat coconut oil diet on metabolism
and adipose tissue steroidogenesis
Serologic evidence of exposure to Ehrlichia spp. in horses in the U.S.
Targeting human breast cancer using a radiolabeled cyclic RGD
peptide
Optimization of a europium-based cytotoxicity assay for equine
mesenchymal stem cells
Examination of Pseudoloma neurophilia infection in the central
nervous system of zebrafish (Danio rerio)
Perinatal growth hormone imprinting of hepatic cytochrome p450
expression in female rats
Identifying antimicrobial resistance patterns in Salmonella enterica
Typhimurium from human and cattle sources
Genome-wide association study seeks connection between stifle
morphology and cranial cruciate ligament disease
Commercialization of an ELISA for Ornithobacterium rhinotracheale
using hemolytic and non-hemolytic isolates
Oxidative stress in stallion semen over time
Identification and characterization of Yersinia ruckeri from farmraised catfish in Mississippi
Role of the gerQ determinant in exosporium structure in Bacillus
anthracis
Role of DDR1 in revascularization and repair after myocardial
cryoinfarction in mice
Alterations in hepatic glucose metabolism following direct exposure
to organochlorine pesticide metabolites
Beneficial effects of probiotic microbes on inflamed gut mucosa in the
rhesus macaque model of HIV infection
Use of the Collaborative Cross to Assay the Impact of Inter-Individual
Variability on TCDD-mediated Signaling
Gut microbiota in shelter felines
Connectional modularity of the mouse inferior colliculus
Construction and expression of full-length canine circovirus molecular
clone
Direct regulation of MLKL by EFhd2 to suppress the necrotic
signaling pathway
LPS stimulated GRK2 expression in response to novel antiinflammatory drugs in raw 264.7 macrophage cells
155
Generation of functional b islet cells from feline adipose-derived
multipotent stromal cells
Phenotypic analysis of adipocyte-specific STAT5 knockout mice
Metagenomic sequencing analysis of Ixodes scapularis virome
Degradation of complement in the presence of Schistosoma mansoni
in human serum
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157
157
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159
159
160
160
161
161
162
162
163
163
164
164
165
165
166
166
167
167
168
Last Name
First Name Poster
#
Ienello
Janse van
Rensburg
Jeffreys
Lauren
Claire
156
157
Roxanna
158
Jimenez
Monica
159
Johnson
Catherine
160
Johnson
Lauren
161
Johnson
Tyler
162
Jones
Melissa
163
Jong
Jordan
Ruby
Mary-ruth
164
165
Joyner
Jalika
166
Kelley-Collier
Parrish
167
Khambholja
Chantelle
168
Khan
Kim
Michael
Eunbee
169
170
Kim
Sohyun
171
Kirby
Klein
Ashley
Hannah
172
173
Kluz
Michael
174
Knebel
Emily
175
Kobluk
Kristi
176
Koenig
Kelsey
177
Koestel
Zoe
178
Kopp
Rosalind
179
Kramer
Caroline
180
Kraneburg
Carol
181
Abstract Title
Page
#
Characterization of blood profiles in free-ranging moose in Minnesota
Do honey bee colonies (Apis mellifera) prefer feed contaminated with
neonicotinoids?
Characterization of the skin microbiome of dogs with strong body
odor and the effect of a spot-on product
Impact of Helicobacter pylori on the infant lung: airway microbiome
and immunomodulation of airway epithelium
MARCKS protein as a therapeutic target in the treatment of equine
asthma
Susceptibility of equine monocyte subsets to equine herpes virus type
I infection
Coagulopathy in Crotalus viridis envenomation, attenuation by carbon
monoxide releasing molecule - 2 in vitro
The Golden Retriever Lifetime Study: factors affecting owner
compliance after the first year of enrollment
Prevalence of feline coronavirus in a specific pathogen-free cat colony
Do you see what I see? Inter-rater reliability of a canine welfare
assessment used in dog breeding kennels
The efficacy of counter conditioning to attenuate signs of fear in dogs
at the veterinary clinic
The effect of chronic Bisphenol A exposure on estrogen-sensitive gene
expression (ER a & ER b, ERR g)
Using in-lab evolution to explain how protozoan parasite Toxoplasma
gondii is such an exceptional generalist
Oil and dispersant induced respiratory distress in Menidia beryllina
Accuracy of PetPace Collar Pulse Rate Compared to Holter Monitor
Heart Rate in Dogs
Isolation and cloning of feline Macrophage Colony Stimulating Factor
for optimizing macrophage differentiation
Expression of TRIM67 in immune stimulated cells
Dysbiosis and bile acid dysmetabolism in dogs with steroid responsive
enteropathy
Evaluation of vertebral articular process dysplasia and the
thoracolumbar myelopathies in pug dogs
Urine for a surprise: Evaluating the use of expanded quantitative urine
culture on feline samples
Genetic knockout screens identify host genes required for zika virus
replication
The effect of oral sodium bicarbonate “milkshake” on serum total
carbon dioxide concentration (TCO2) in horses
Is dietary exposure to bisphenol A causing phenotypic changes in
dogs?
Evaluation of bone marrow aspiration methods to increase yield and
reduce time for therapeutic dose expansion
Rates and Risk Factors for Surgical Site Infections at the Ohio State
University Veterinary Medical Center
Contraception of feral cats with phage-based vaccines targeting
gonadotropin releasing hormone (GnRH)
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Last Name
First Name Poster
#
Kuball
Kristin
182
Kucera
Cecilia
183
Kumar
Anisha
184
Kustasz
Lauren
185
Lake
Lamb
Bathilda
Holly
186
187
Landis
Casey
188
Landreth
Victoria
189
Lantz
Maya
190
Larrabee
Shannon
191
Laurence
Hannah
192
LaVine
Danielle
193
Lawnichak
Tyler
194
Ledesma
Eric
195
Lee
Amber
196
Li
Meng
197
Lieberman
Seth
198
Lin
Dian Dian
199
Lin
Megan
200
Lind
Lori
201
Linn
Sarah
202
Livengood
Kristen
203
Loeschel
Courtney
204
Long
Mackenzie
205
Abstract Title
Page
#
Using Cold Atmospheric Plasma as an Alternative Treatment for
Chronic, Antibiotic Resistant Wounds
Determining improved prognostic indicators in equine large colon
volvulus
Ataxic Horses Display Quantifiable Gait Abnormalities Determined
By Videography
Optimization of ultrasound settings for better determination of
cystolith size in vitro
Equine PCV, Erythrocyte Potassium and Transferrin Correlation
Evaluation of the roles of plasmin and CCL20 on nerve damage in an
inflammatory M. leprae infection model
Characterization of a co-culture model for observation of Francisella
tularensis infection
Exploration of tongue muscle pathology in canine degenerative
myelopathy
The effects of pet ownership on academic achievement among
veterinary students at St. George’s University
Discovery of a genetic mutation that causes sudden cardiac death in
toy Manchester terriers
The architecture of the Flavivirus 3’ untranslated region, pathogenic
RNA production, and viral cytopathicity
Yeast surface display library to yield prospective homing protein to
the ischemic myocardium
Influence of donepezil on the development of Toxoplasma gondii and
its potential relationship to Alzheimer’s
Validation of putative chromosome 14 risk alleles for osteochondrosis
in Standardbred horses
Examination of insectivorous bats (Molossus molossus) in Grenada
for trematodes and Neorickettsia DNA
Management practices and disease perceptions among Minnesota
backyard flock owners following a HPAI outbreak
Characterization of a Sox2-expressing subpopulation of cells in canine
lymphoma
Two methods of Clostridium difficile toxin purification for detection
by MALDI-TOF MS
Progression of vertebral bone pathology in mucopolysaccharidosis VII
dogs from birth to skeletal maturity
The effects of tongue injection of CTB-SAP on ventral hypoglossal
motor neurons: a novel model of ALS
Experimental modeling of the nonspecific protective effects with
measles virus vaccination
Biofilm formation by mastitis-producing Staphylococcus aureus and
inhibition by 2-aminoimidazole compounds
Evaluation of methods to preserve field samples for detecting elephant
endotheliotropic herpesviruses
Induced GI tract microbial dysbiosis does not accelerate disease
progression in SIV-infected Asian macaques
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183
184
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185
185
186
186
187
187
188
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189
189
190
190
191
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192
192
193
Last Name
First Name Poster
#
Lopez
Ariana
206
Lopez
Melissa
207
Lou
Jon
208
Lovering
Lucyshyn
Samantha
Danica
209
210
Ludwig
Allison
211
Ludwig
Lundquist
Luu
Latasha
Matthew
Savannah
212
213
214
Lyakhova
Tatyana
215
Mack
Tyler
216
Madden
Kelsey
217
Maeroff
Jacqueline
218
Maier
Kaitlin
219
Malcolm
Elizabeth
220
Mangosing
Sara
221
Mann
Olivia
222
Manning
Hannah
223
Mariampillai
Anujah
224
Marinoff
Jacqueline
225
Marshall
Martin
Breanna
Alison
226
227
Martinez
Martinez
Aileen
Samantha
228
229
Mason
Caitlin
230
Masters
Allison
231
Abstract Title
Page
#
Effect of dietary antioxidant deficiency on canine central vision: a
pilot study
Characterization of vascular remodeling in the canine brain following
Traumatic Brain Injury
193
b-cell glucagon-like peptide-1 receptor improves islet morphology
after vertical sleeve gastrectomy in mice
The role of Borrelia burgdorferi infection in canine myocarditis
Sudden acquired retinal degeneration syndrome in Western Canada: a
retrospective (2002-2016)
Evaluating human pluripotent stem cell-based photoreceptor
transplantation in rodent models
Innate T cells on the frontlines of host immunity and defense
Host immunomodulation by parasitic nematode exosome-like vesicles
Profile of gap junction intercellular communication in canine
mammary carcinoma
Soil-borne protozoans enable persistence of facultative intracellular
pathogens in the environment
Immediate treatment of hyperketonemia ameliorates negative impacts
on milk production but not comorbidity risk
NSAIDs inhibit wound healing in intestinal and reproductive
epithelial cell lines
Comparison of lymphocyte count and proviral load in beef and dairy
cattle infected with bovine leukemia virus
Investigation of sarcolipin as a cause of recurrent exertional
rhabdomyolysis in horses
Radiographic and echocardiographic assessment of left atrial size in
dogs with mitral regurgitation
Generation of Infectious Bronchitis Virus (IBV) M41 cDNA clones
for use in chimeric IBV virus construction
Characterization of N-linked glycan profiles in bat gastrointestinal
tissues specific to influenza A viruses
Using behavior to establish a timed oxytocin protocol for estrous
suppression in mares
Biomonitoring the effects of BPA and its metabolite BPAG during
fetal life
Optimization of extraction and quantitation of cfDNA from equine
plasma
Characterization of a calcium-binding protein from Toxoplasma gondii
Adoption of recommended hand hygiene practices to protect public
health at agricultural fairs
Gait Assessment after peripheral nerve injury using Rotarod
Evaluation of novel drug therapies against human breast and canine
mammary carcinomas
Native and recombinant PD-1, PD-L1, and PD-L2 variants for study
of porcine circovirus associated disease
Effects of anti-inflammatory glucocorticoids on hemodynamics and
cardiac function in clinically healthy dogs
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201
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202
203
203
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206
Last Name
First Name Poster
#
Mattia
Michael
232
May
Emily
233
McClung
George
234
McGeehan
Megan
235
McLean
Victoria
236
McMahon
Rachel
237
McMullin
McSweeny
Medland
Alan
Ian
Julia
238
239
240
Melhado
Ashlee
241
Meltzer
Melvin
Rebecca
Rachel
242
243
Mendoza
Gerardo
244
Mercer
Stephen
245
Mertens
Micaela
246
Middlebrooks
Miller
Sarah
Travis
247
248
Millman
Mills
Zachary
Kelly
249
250
Mironovich
Melanie
251
Mitchell
Elspeth
252
Mitchell
Freelie
253
Moorhead
Mosca
Wil
Leandra
254
255
Moses
Amber
256
Moyer
Muller
Elizabeth
Stefanie
257
258
Abstract Title
Page
#
Fate of exosporium proteins BclA, BclB, and BxpB in germinating
Bacillus anthracis spores
Antigen presenting cells promote protective immunity in catfish
exposed to live Edwardsiella ictaluri vaccines
Immunophenotype comparison of equine bone marrow-derived and
synovial membrane-derived mesenchymal stem cells
Characterization of tertiary lymphoid organs in a natural model of
Multiple Sclerosis
Morphology of peritoneal endometriotic lesions using an in-vivo
mouse model of retrograde menstruation
Obesity promotes breast tumor development and metastases in an
immunocompetent mouse model
Biomechanical Properties of the Porcine Optic Nerve
Chronic intermittent hypoxia increases breathing instability in mice
Sur1 and TRPM4 expression in the spinal cord of dogs with
progressive myelomalacia
Pathophysiologic environmental factors diminished efficacy of
ceftiofur and penicillin with gentamicin
Investigating the role of Ndr kinase in retina development
Oviduct morphology and Prolactin receptor expression in N.
American River Otters and Asian Small Clawed Otters
A single mutation in the vaccinia virus RNA polymerase inhibits
multiple host restriction factors
Domains of CD163 involved in infection of Type I and Type II
PRRSV
Monitoring bull behavior to determine characteristics of bull
dominance and the role in social ranking
T. Gondii Infection and Nurr1-null Genotype Affect Spatial Learning
Effect of a supplement containing curcumin on lameness due to
osteoarthritis or degenerative disease
Validating the Cell of Origin of T-Zone Lymphoma
Utility of ATP-bioluminescence measured by a luminometer to
monitor hygiene of milk replacer for dairy calves
STAT3 activation via the IL6/gp130 pathway in sepsis-related and
hyperinsulinemic laminitis
Antibiotic alternatives for the reduction of morbidity and mortality in
veal calves
Optimization of DNA Vaccine Targeting GnRH Receptor for Animal
Contraception
Gender differences in protein detection of swine muscle and fat tissue
Comparative immunogenicity of attenuated Salmonella vaccine
strains dependent on parental origin and genotype
Changes in periarticular bone secondary to osteoarthritis in
chikungunya-infected and naive mice
Evaluation of intradermal testing in five horses
Characterizing interactions between location, land use, and domestic
dog health in Panamanian communities
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207
208
208
209
209
210
210
211
211
212
212
213
213
214
214
215
215
216
216
217
217
218
218
219
219
Last Name
First Name Poster
#
Murray
Olivia
259
Mustonen
Allison
260
Myers
Laurel
261
Nagel
Nguyen
Jonathan
Chrystal
262
263
Nguyen
Niel
Steven
Kayla
264
265
Nietlisbach
Nicole
266
Nilles
Jacob
267
Oelfke
Thomas
268
Ofer
Oldeschulte
Oren
James
269
270
Olney
Oltman
Erika
Wesley
271
272
Onaga
Robert
273
Ondera
Caitlin
274
Pador
Page
Sean
Lauren
275
276
Pannone
Stephen
277
Patterson
Delaney
278
Pérez Irizarry
Cristian
279
Perier
Nadege
280
Perry
Laura
281
Phillips
Elsie
282
Piegols
Hunter
283
Pike
Courtney
284
Abstract Title
Page
#
Attempt to confirm absence of Avian Bornavirus in a previously
shedding flock of cockatiels (N. hollandicus)
How to smoke your bacon: An investigation on the effects of
environmental inhalants in a porcine model
Characterizing infection of Jamaican fruit bats with bat-derived
influenza-like viruses, HL17N10 and HL18N11
Characterization of the Dog Erythrocyte Antigen 1 protein and gene
Effect of Hypoxic Preconditioned Wharton Jelly Mesenchymal Stem
Cell Exosomal miRNA on Hypoxic Injured Neurons
Quality and accuracy of the smartphone ECG in cats
The growing feather as a model for novel evaluations of Marek’s
disease genetic resistance
Thermal nociception and gastrointestinal function following
administration of Simbadol in rats
Discovery of common genetic risk loci for canine cruciate ligament
rupture by validation GWAS
A proposed method for functional quantitation of carpal joint laxity in
canine mucopolysaccharidosis type I
Smartphone-based ECG vs. standard computer-based ECG in dogs
Development of large animal models for a compound heterozygous
human disease using CRISPR/Cas9
Interleukin-27 receptor expression in murine intestinal organoids
Seasonal variation of 25-hydroxy-vitamin D in two captive Eastern
black rhinoceros (Diceros bicornis michaeli)
Licorice root: a phytoestrogen supplement with neuroprotective
effects on cognition
Therapeutic intervention of hantavirus disease
Efficacy of a new diagnostic procedure, the Mini-FLOTAC, in the
diagnosis of canine Giardia infection
In vivo and in vitro evaluation of mesenchymal stem cell therapy for
primary osteoarthritis
Assessment of cellular response to synovial joint fluid in osteoarthritic
and healthy dogs
Effects of intravenous fluid therapy on serum osmolarity in
hospitalized dogs
The impact of mastitis on maternal behavior in dairy cows: a pilot
study
Neutralization of chlorhexidine-containing products in a clinical hand
hygiene efficacy study
Role of Mouse Sca1+ Lung Mesenchymal Stem Cells in Bacterial
Pneumonia
Investigation of novel chemotherapeutic therapies for feline oral
squamous cell carcinoma
Venturing into the unknown: characterization of the aerobic cultivable
bacteria in wounds of wildlife species
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222
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223
224
224
225
225
226
226
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228
228
229
229
230
230
231
231
232
232
Last Name
First Name Poster
#
Pinnell
Erin
285
Pires
Poole
Porter
Elena
Tyler
Stephanie
286
287
288
Powers
Prado-Sanchez
Jonathan
Evymarie
289
290
Price
Emma
291
Pruett
Katherine
292
Pulczinski
Jairus
293
Putman
Ramirez
Berrios
Ramos
Ashley
Karen
294
295
Meghan
296
Randall
Chelsea
297
Rasche
Brittany
298
Rash
Reinhard
Morgan
Natalie
299
300
Reising
Abby
301
Rettkowski
Rice
Rebecca
Deja
302
303
Rice
Meredith
304
Richardson
Rimmelin
Ella
Alesha
305
306
Roberson
Rachel
307
Rodenbaugh
Rose
Cassandra
Hailey
308
309
Rose
Tierra
310
Roseman
Brandi
311
Ross
Taylor
312
Abstract Title
Page
#
Optimal decellularization produces synovial extracellular matrix
scaffold for in vivo cartilage regeneration
Insights into homologous recombination DNA repair in mammals
Production of recombinant ferret cytokines in multiple cell substrates
Environmental Reservoirs of Francisella tularensis: Mechanisms for
Transmission of Tularemia
Anti-Toxoplasma efficacy of green algae extracts
Nickel homeostasis and bacterial pathogenesis: Examining the NikR
regulatory system in Brucella abortus
Characterization of the role of type I and type III interferons in acute
and chronic disease
Examining the relationship between affective state and social rank in
group-housed gestating sows
Maternal markers of inflammation and oxidative stress following air
pollution exposure during pregnancy
Cytochrome expression in bovine tissues and cells
Establishing baseline EEG parameters in reptiles
233
Biosensing of biofilm: detecting volatile organic compounds using a
dog’s nose
Evaluating body condition score with body weight on serum drug
levels of phenobarbital and potassium bromide
Environmental awareness: Recreating the natural tissue niche in vitro
for cancer research
The effect of cryopreservation on the size of the canine meniscus
Structural modifications to K+ channel, Kv1.1, in a wild rodent
resistant to scorpion venom
Demonstrating C. botulinum neurotoxin heavy chain cytosolic
localization via modified Kirby-Bauer assays
Development of a risk assessment map of fish kills across Minnesota
Mast cells in allergies and in asthma- does particulate matter (PM)
activate mast cells?
Active veterinary surveillance of emerging Ixodes scapularis tick
populations in Michigan
Competitive exclusion of Campylobacter jejuni in the chicken gut
Disparate virulence in two plaque variants of Theiler’s murine
encephalitis virus, DA strain
Endothelial colony forming cells as treatment for equine distal limb
wounds
Sonoclot evaluation of whole blood coagulation in chickens
Mutant XYZ is associated with hyper-activation of mTOR and T cell
senescence
Loss of parvalbumin-immunoreactive interneurons in epileptic
California sea lions
Parkinson disease modeling in parkin-deficient mice: testing synthetic
mitochondrial-targeted antioxidants
Investigating the role of Delta-like ligand 1, a Notch signaling ligand,
in breast cancer metastasis
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235
236
236
237
237
238
238
239
239
240
240
241
241
242
242
243
243
244
244
245
245
246
246
Last Name
First Name Poster
#
Rowbotham
Nicole
313
Rowe
John
314
Russell
Sablotny
Ellen
Elizabeth
315
316
SanabriaOjeda
Sato
Laura
317
Yu
318
Saturno
Julia
319
Schacher
Schade
Scheffe
Schenk
Schettler
Michael
Seana
Gretchen
Alex
Michael
320
321
322
323
324
Scheuermann
Logan
325
Schoenberger
Jenna
326
Schrock
Kelly
327
Schulte
Rachael
328
Schulze
Laura
329
Schuster
Chelsea
330
Scott
Jenna
331
Seelman
Amanda
332
Senft
Tyler
333
Shearin
Abigail
334
Si
Siddons
Catherine
Giles
335
336
Siegrist
Sirochman
Melissa
Anna
337
338
Sirois
Alexis
339
Abstract Title
Page
#
Involvement of increased CB1R activation in altered social play
induced by developmental chlorpyrifos exposure
Intestinal epithelial cells NF-kB regulates host response to ingestion
of low doses of cadmium
NLR inflammasome recognition of Clostridium difficile
Effect of litter type on survival of Tritrichomonas foetus in feline
feces
Effects of the JAK-inhibitor AZD1480 on activated dorsal root
ganglia and itch behavior in mice
Behavioral Temperament of rhesus macaques (Macaca mulatta) and
the association with gut microbiota
Temperature as a confounding variable in oncolytic virotherapy for
canine melanomas
Effects of GM-CSF on critically ill dog’s immune cell function
Investigation of the MDR1 Gene in Reptile Species
Knock-down of tmem150a increases cytokine release
Investigation of post-sterilization hyphema in shelter cats
No evidence for sexual dimorphism of nitric-oxide (NO) mediated
vascular control in rat skeletal muscle
Feasibility of viable synovial engineered transplants for paracrine
influence on cartilage regeneration
Effects of calorie restriction on radiation resistance of reserve
intestinal stem cells
Molecular etiology of intrahepatic portosystemic shunts in the Nova
Scotia Duck Tolling Retriever
Paraventricular nucleus neurons target nucleus tractus solitarii via
CRH receptors and oxytocin in hypoxia
Down stream effects of bovine respiratory disease complex on
myocardial gene expression
Intraperitoneal ethanol injection for humane euthanasia in zebra
finches (Taeniopygia guttata): a pilot study
Effects of deworming and vaccination on seroresponse to BVDV1 and
BHV1 in stocker calves
Identification of Arginine Vasopressin producing neurons in the
amygdala of domestic cats
Two trials to investigate techniques for canine Vitamin D
supplementation
Adipose AKT2 controls circulating free fatty acids and systemic
insulin resistance
Modeling breast cancer with patient-derived orthotopic xenografts
Schistosomes recruit host regulatory proteins to resist complement
attack
Development of differing complex microbiota in CD1 mice
Is there evidence for canalithiasis or cupulolithiasis as an etiology of
canine idiopathic vestibular disease?
Examination of genetic variation in CD18 of cattle as a marker for
resistance to respiratory disease
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250
251
251
252
252
253
253
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255
256
256
257
257
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258
259
259
260
Last Name
First Name Poster
#
Slabaugh
Taylor
340
Slimack
Katie
341
Smit
Michelle
342
Smith
Smith
Carolyn
Courtney
343
344
Smith
Katelin
345
Smith
Katherine
346
Smith
Kayla
347
Smith
Mallory
348
Smith
Richard
349
Smith
Smyth
Samuel
Lauren
350
351
Snyder
Kristin
352
Soltis
Emily
353
Sommer
Samantha
354
Sorenson
Sorg
Juli
Janna
355
356
Stabler
Emily
357
Stelly
Philip
358
Stenger
Robert
359
StenkampStrahm
Stephen
Chloe
360
Alexa
361
Stephenson
Olivia
362
Stern
Daniel
363
Stilin
Allison
364
Abstract Title
Page
#
Frequency and complexity of arrhythmias in rhesus macaques with
and without hypertrophic cardiomyopathy
The role of lidocaine and meloxicam in pain management for bull
castrations
Genetic investigations of the AMH and AMHR2 genes in canine
persistent Mullerian duct syndrome
Frequency of forelimb weight shifting in normal horses
Comparison of molecular methods in the characterization of
Malassezia on the skin of healthy and allergic dogs
Eosinophil activity is not altered by respiratory syncytial virus
infection in the cotton rat
Bacterial killing ability of vampire bats and associations with
individual body condition and feeding ecology
Tricaine methane sulfonate (MS-222) anesthesia in fishes:
Relationship to thermal divergence
In vitro effects of the chemotherapeutic agent water-soluble paclitaxel
on canine mast cell tumor cell lines
VP22 transport from differentiated Neuro2A cells in microfluidic
chambers to epithelial cells and vice versa
Do Noviplex cards have a future in veterinary clinics?
A Comparison of Lyme disease Strain Diversity in Michigan Island
and Mainland Sites
Why won’t you eat me? Investigating melanomas’ unique resistance
to phagocytosis through RNAi
The effect of prenatal alcohol exposure on respiratory regulation
during sleep in the early postnatal period
A new incapacitance meter for the measurement of static weightbearing forces in the hind limbs of mice
Creation of a mouse model with ‘equinized’ gut microbiota
The effect of more liberal milk allowance in the first week of life on
performance of preweaned dairy calves
Benefits of a therapeutic horseback riding program on veterans
suffering from PTSD or traumatic brain injury
260
gd T Cells play a critical role in the inflammatory response to
Paramyxovirus infection in mice
Helminths in mountain gorillas: conservation through intestinal
parasite monitoring
Microbial community structures in E. coli O157 shedding and nonshedding dairy cattle; a preliminary analysis
The effects of natural and synthetic retinoid therapy on feline
squamous cell carcinoma cell lines
Characterization of exosome miR-9 expression in canine
osteosarcoma
Use of susceptible and avirulent E. coli to reduce antimicrobialresistant coliforms in milk-fed calves
Discovery of the causal variant for acquired laryngeal paralysis
polyneuropathy in the dog
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267
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269
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270
271
271
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272
Last Name
First Name Poster
#
Storms
Suzanna
365
Stovall
Kelsie
366
Strathe
Erin
367
Streu
Shayna
368
Stricklin
Olivia
369
Stromberg
Stephanie
370
Strumpf
Sundaram
Alyssa
Ayswarya
371
372
Swanson
Tracy
373
Tageant
Connor
374
Tamas
David
375
Tang
Tang-Wing
Karena
Cassandra
376
377
Tarlowe
Stephanie
378
Tatro
Melissa
379
Taupier
Rachel
380
Tavella
Vincent
381
Taylor
Michelle
382
Tenborg
Elizabeth
383
Thompson
Alexis
384
Timonin
Touitou
Mary
Florian
385
386
Tran
Kim
387
Trearchis
Deanna
388
Tuncay
Mete
389
Abstract Title
Page
#
The effects of human interactions on gastrointestinal parasites in
Mediterranean mouflon in southern France
Transient Receptor Potential Melastatin 4 channel enhances
osteogenesis of rat dental pulp stem cells
A systematic review of the literature to identify and quantify host and
vector competence and abundance of JEV
A questionnaire-based study on predisposing and risk factors for
gastric dilatation volvulus in dogs
Effects of caffeine on injured and uninjured intervertebral discs in a
whole organ culture model
Genotype and phenotype of sudden acquired retinal degeneration
syndrome (SARDS) in dachshunds
Antimicrobial properties of human mesenchymal stem cells
Deciphering the interactions between feline mesenchymal stem cells
(MSCs) and CD8+ T cells in vitro
Defining the relationship between female sex hormones and
myocardial fibrosis in aortic-banded mini-swine
Vaccination of small ruminants against Haemonchus contortus using
fecal antigens
The effects of inflammation on the pharmacokinetics of flunixin in
steers using in vivo ultrafiltration
Morphometric study of feline amygdala using serial sections
Aedes Aegypti mosquito transmission and the development of STAT2
KO hamster animal model for Zika virus
Diagnostic accuracy of Keto-Test milk strips for cowside detection of
elevated milk b-hydroxybutyrate
Marek’s disease infectivity in genetically susceptible and resistant
chickens
Effects of Bedoukian compound concentrations on gastrointestinal
nematode development in sheep
Control of lupus nephritis by manipulating gut microbiota during
active disease
Evaluation of health in Texas tortoises via multiple patient-side
analytes
How does Toxoplasma gondii deliver effector proteins that control
host cell functions
Evaluation of refractometry to detect failed transfer of passive
immunity in pre-weaned beef calves
Iodixanol is an effective cryoprotectant for mouse spermatozoa
Development of less invasive functional assessments of malignant
hyperthermia (MH) genotypes
In vivo evaluation of a novel HER2 inhibitor, lapatinib, in
osteosarcoma
Prevalence of Failure of Passive Transfer, Dehydration, and Health
Outcomes in Veal calves on Day of Arrival
A Novel Mechanism that Regulates the Surface Levels of NKp46 on
Natural Killer Cells
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280
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283
284
284
285
Last Name
First Name Poster
#
Turinski
Tiffani
390
Turner
Zachary
391
Vadala
Clara
392
van Batavia
Ashley
393
van den
Hoogen
Varley
Marjan
394
Ashley
395
Velazquez
Eric
396
Vella
Gianna
397
Vermillion
Meghan
398
Vetter
Nicholas
399
Vijan
Stephanie
400
Vincent
Emily
401
Vonderohe
Caitlin
402
Voss
Wahl
Brittany
Alexandra
403
404
Walden
Katherine
405
Walker
Anna
406
Walker
Ashley
407
Walker
Meghan
408
Wanner
Nicole
409
Warcholek
Stanislaw
410
Ward
Kinnon
411
Ware
William
412
Watson
Claire
413
Abstract Title
Page
#
Characterization of the fecal microbiome from EHEC positive and
digital dermatitis negative beef cattle
Fescue toxicosis in grazing beef cattle: impact of environmental
temperature and humidity
Carnoy’s vs. Formalin: a comparison of two fixatives in colonic
samples from weaned piglets
Survey of infectious diseases in free-ranging Chilean Humboldt
penguins (Spheniscus humboldti)
Development of a Canine Functional Questionnaire for Assessing
Rehabilitation Outcomes
Comparison of virus isolation from fabric for use in herd surveillance
of viral bovine respiratory disease
Commensal Enterobacteriaceae Drive Variable Salmonella
Colonization Resistance of Mice from Different Vendors
Using human cell-specific data and correlation to cell proportion to
predict pig neutrophil-specific genes
Estriol protects female mice against influenza A virus by reducing
pulmonary inflammation
Rodent Models of Ozone-Induced Eosinophilic Rhinitis: Species- and
Strain-Dependent Variations
Association of progestogens with inflammation and immunity in
critically ill foals
Field research analysis of free-roaming cat populations on The Ohio
State University’s campus
The effect of antibiotic-free management on the environmental impact
of swine production
Purification of hantavirus cap snatching endonuclease
Florfenicol resistance in chloramphenicol-resistant canine isolates of
Staphylococcus pseudintermedius
Effects of src homology 2 containing inositol 5-phosphatase (SHIP)
Inhibitors on canine osteosarcoma cells
Assessment of gastrointestinal nematode parasitism in dairy calves
housed in calf hutches
Ambulatory ECG, heart rate variability and cardiac stress in cats with
subclinical hypertrophic cardiomyopathy
Development of a next-generation sequencing method for veterinary
forensics
Impact of a paternally inherited allele for polysaccharide storage
myopathy on muscle pain and myodegeneration
Tumor Microenvironment Analysis via High Throughput Proteomics
after Poly(I:C)ZnO Treatment
The effects of ischemic stroke on cardiac inflammation in
normotensive rats
The effects of target distance and gun type on accuracy of 8cc darts
used in remote drug delivery systems
Deletion of liver fatty acid binding protein increases liver fat, bone
marrow fat and bone mineral density
285
55
286
286
287
287
288
288
289
289
290
290
291
291
292
292
293
293
294
294
295
295
296
296
297
Last Name
First Name Poster
#
WegnerClemens
Weigel
Erik
414
Ellen
415
Werner
Melissa
416
Weyna
Alisia
417
Whisenant
Katrijn
418
Whittmore
Wickware
Jerrianne
Taylor
419
420
Wierenga
Lauren
421
Wilcox
Parker
422
Williams
Courtney
423
Williams
Jessica
424
Witschen
Patrice
425
Wong
Stephanie
426
Wright
Courtney
427
Wronski
Yang
Sarah
Stephanie
428
429
Yang
Young
Steven
Kimberly
430
431
Zabrecky
Kristin
432
Zhou
Xueying
433
Zlotnick
Marta
434
Zorn
Chelsea
435
Zuber
Emily
436
Zygelyte
Emilija
437
Abstract Title
Page
#
Characterizing ischemic wound healing in jejunal epithelial cells in
porcine neonates and juveniles
Diallyl trisulfide inhibits epithelial cell adhesion molecule signaling in
hepatocellular cancer
Accessing Potential Causes of Repeat Breeder Dairy Cows through
Cytology, Bacteriology, and Histopathology
Feline ocular and periocular squamous cell carcinoma: a retrospective
study of 374 cases
Targeting glycolysis reduces inflammation in a murine models of
rheumatoid arthritis and Crohn’s disease
Changes in the equine gut microbiome associated with hospital stay
Immortalization of Feline Mesenchymal Stem Cells with Feline
Telomerase and Novel Splice Variant Isoforms
Exploring the cellular origin of myelin repair in a chronic model of
demyelination and remyelination
Efficacy and safety of topical nitric oxide donors on decreasing
intraocular pressure in healthy canine eyes
Identification of Aeromonas hydrophila virulence genes providing
complement resistance
Effects of microaggregate filters on canine erythrocyte viability and
morphology
Hyaluronan processing and function in the progression of breast
cancer
Deleterious glutathione-S-transferase-pi (GSTP1) polymorphisms may
contribute towards Boxer dog lymphoma risk
Detection of influenza A virus on inanimate surfaces at agricultural
fairs
Host-oriented inhibitors of Lassa fever virus budding
Prenatal infection with Listeria monocytogenes and risk for
neurodevelopmental defects in pregnant mouse model
Antimicrobial Properties of Equine Mesenchymal Stem Cells
Phenotypic evaluation of multicopy single-stranded DNA mutants to
determine functional regions
Gut microbiota analysis of an Alzheimer’s Disease transgenic rat
model
Characterization of canine bone marrow mesenchymal stem cells
towards development of brain repair strategies
Comparison of infectivity of Zika virus strains in primary adult
murine neurons
Isolation of uniquely recognized salivary gland antigens to interfere
with feeding performance of ixodid ticks
Placental abnormalities and fetal growth restriction as a consequence
of preconception male alcohol exposure
Demethylase inhibitor reduces equine herpesvirus-1 expression in
vitro
297
56
298
298
299
299
300
300
301
301
302
302
303
303
304
304
305
305
306
306
307
307
308
308
309
Abstract Index by Research Area
The research areas are as follows:
Biomedical Engineering and Regenerative Medicine
Cell Biology and Cancer Cell Biology
Developmental Biology
Endocrinology
Epidemiology/Public Health
Genetics/Genomics
Hematology
Infectious Disease and Immunology
Molecular Biology and Biochemistry
Pathology
Pharmacology/Toxicology
Biomedical Engineering and Regenerative Medicine
Last Name
First Name Poster
#
Bollman
Mary Kate
42
Byerley
Condrey
Sydney
Jillian
48
77
Fleming
Payge
108
Gavitt
Ashley
116
Haffner
125
Haight
Abigail
(Abbie)
Kimberly
Harris
Elizabeth
134
Hughes
Peyton
152
Joyner
Jalika
166
Kim
Eunbee
170
Kopp
Rosalind
179
Kumar
Anisha
184
LaVine
Danielle
193
Lopez
Melissa
207
128
Abstract Title
Page
#
Application of additive manufacturing technology and stem cell
recruitment to improve osseointegration
Effect of mesenchymal stem cell conditioned media on TGF-β1
exposed equine fibroblasts
Nexelom cellometer live cell and viability verification and error
evaluation for the canine stem cell assay
Custom 3D-printed drill guides for surgical stabilization of the canine
cervical spine - a cadaveric study
Investigating novel small compounds and their effect on inflammation
and chondrogenesis in joints
A porcine model of vascular cognitive impairment for dementia
prevention
Optimization of a europium-based cytotoxicity assay for equine
Generation of functional β islet cells from feline adipose-derived
multipotent stromal cells
The efficacy of counter conditioning to attenuate signs of fear in dogs
at the veterinary clinic
Accuracy of PetPace Collar Pulse Rate Compared to Holter Monitor
Heart Rate in Dogs
Evaluation of bone marrow aspiration methods to increase yield and
reduce time for therapeutic dose expansion
Ataxic Horses Display Quantifiable Gait Abnormalities Determined By
Videography
Yeast surface display library to yield prospective homing protein to the
ischemic myocardium
Characterization of vascular remodeling in the canine brain following
Traumatic Brain Injury
111
57
114
129
144
148
153
154
157
166
173
175
180
182
187
193
Last Name
First Name Poster
#
Ludwig
Allison
211
Martinez
McClung
Aileen
George
228
234
McMullin
Nguyen
Alan
Chrystal
238
263
Nguyen
Ofer
Pannone
Steven
Oren
Stephen
264
269
277
Pinnell
Erin
285
Rash
Roberson
Morgan
Rachel
299
307
Scheuermann
Logan
325
Smith
Sommer
Carolyn
Samantha
343
354
Sundaram
Ayswarya
372
van den
Hoogen
Wierenga
Marjan
394
Lauren
421
Yang
Steven
430
Anderson
Jessica
8
Arica
Andrea
9
Au
Gina
12
Bartholomew
Kristen
24
Becher
Jessica
27
Belling
Binstock
Carolyn
Jonah
28
34
Bloom
Calle
Christopher
Jasmine
40
53
Abstract Title
Page
#
Evaluating human pluripotent stem cell-based photoreceptor
transplantation in rodent models
Immunophenotype comparison of equine bone marrow-derived and
synovial membrane-derived mesenchymal stem cells
Effect of Hypoxic Preconditioned Wharton Jelly Mesenchymal Stem
Cell Exosomal miRNA on Hypoxic Injured Neurons
In vivo and in vitro evaluation of mesenchymal stem cell therapy for
primary osteoarthritis
Optimal decellularization produces synovial extracellular matrix
scaffold for in vivo cartilage regeneration
Endothelial colony forming cells as treatment for equine distal limb
wounds
Feasibility of viable synovial engineered transplants for paracrine
influence on cartilage regeneration
A new incapacitance meter for the measurement of static weightbearing forces in the hind limbs of mice
Deciphering the interactions between feline mesenchymal stem cells
(MSCs) and CD8+ T cells in vitro
Development of a Canine Functional Questionnaire for Assessing
Rehabilitation Outcomes
Exploring the cellular origin of myelin repair in a chronic model of
demyelination and remyelination
Antimicrobial Properties of Equine Mesenchymal Stem Cells
196
Cell Biology and Cancer Cell Biology
The impact of a cancer-specific diet on inflammation and side effects
associated with chemotherapy in dogs
Investigating the effects of EZH2 inhibition in canine B-cell
lymphoma
Combinatory radiotherapy and cellular immunotherapy enhances tumor
cell killing
Novel in vitro model system of canine degenerative myelopathy using
direct conversion of skin fibroblasts
Hormonal effects on adenoma development in a rat model of human
familial colon cancer
Col3’s roles in the fibrotic response: therapeutic strategies to engineer
regenerative and tumor environments
The role of Hus1 impairment in non-small cell lung cancer
tumorigenesis
58
204
207
209
222
222
225
229
233
240
244
253
262
267
276
287
301
305
94
95
96
102
104
104
107
110
117
Last Name
First Name Poster
#
Carlson
Ariel
55
Cleveland
Ashley
74
Courville
Cox
Emily
Amanda
78
79
Cutcliffe
Ross
83
Ehrlich
Allison
99
Geisler
Jennifer
118
Girens
Renee
121
Hargis
Jessie
133
Hendrikx
Herrold
Lysanne
Emily
140
143
Huff
Jaimie
150
Hughes
Courtney
151
Hunt
Kucera
Janna
Cecilia
153
183
Lieberman
Seth
198
Lopez
Ariana
206
Luu
Savannah
214
Madden
Kelsey
217
Marshall
Martinez
Breanna
Samantha
226
229
McMahon
Rachel
237
Medland
Julia
240
Meltzer
Millman
Mironovich
Rebecca
Zachary
Melanie
242
249
251
Patterson
Delaney
278
Abstract Title
Page
#
The effects of metformin and resveratrol on canine hemangiosarcoma
cell lines
Anatomy of the lower respiratory system of the African Grey Parrot
(Psittacus erithacus erithacus)
Role of inhibitor of differentiation (Id) proteins in fibrosis of the human
cornea
Cancer Stem Cell-Targeted Vaccine Induces Antibodies Against
Conserved Vaccine Antigens
Breast cancer cell growth mediated by EGFR/HER1 expression is
regulated by p53 suppressor function
miR146a is an endogenous regulator of both hematopoiesis and bone
mass
Calculation of body surface area using computed tomography-guided
modeling in dogs and cats
Targeting human breast cancer using a radiolabeled cyclic RGD
peptide
Role of DDR1 in revascularization and repair after myocardial
cryoinfarction in mice
Direct regulation of MLKL by EFhd2 to suppress the necrotic signaling
pathway
LPS stimulated GRK2 expression in response to novel antiinflammatory drugs in raw 264.7 macrophage cells
Determining improved prognostic indicators in equine large colon
volvulus
Characterization of a Sox2-expressing subpopulation of cells in
canine lymphoma
Effect of dietary antioxidant deficiency on canine central vision: a pilot
study
Profile of gap junction intercellular communication in canine
mammary carcinoma
NSAIDs inhibit wound healing in intestinal and reproductive epithelial
cell lines
Evaluation of novel drug therapies against human breast and canine
mammary carcinomas
Obesity promotes breast tumor development and metastases in an
immunocompetent mouse model
Sur1 and TRPM4 expression in the spinal cord of dogs with
progressive myelomalacia
Investigating the role of Ndr kinase in retina development
STAT3 activation via the IL6/gp130 pathway in sepsis-related and
hyperinsulinemic laminitis
Assessment of cellular response to synovial joint fluid in osteoarthritic
and healthy dogs
118
59
127
129
130
132
140
149
151
157
160
162
165
166
167
182
189
193
197
199
203
205
209
210
211
215
216
229
Last Name
First Name Poster
#
Piegols
Hunter
283
Ramos
Meghan
296
Rasche
Brittany
298
Ross
Taylor
312
Saturno
Julia
319
Si
Smith
Catherine
Mallory
335
348
Snyder
Kristin
352
Stephen
Alexa
361
Stephenson
Stovall
Olivia
Kelsie
362
366
Stricklin
Olivia
369
Tang
Tran
Karena
Kim
376
387
Tuncay
Mete
389
Walden
Katherine
405
Warcholek
Stanislaw
410
WegnerClemens
Weigel
Erik
414
Ellen
415
Wickware
Taylor
420
Witschen
Zhou
Patrice
Xueying
425
433
Abstract Title
Page
#
Investigation of novel chemotherapeutic therapies for feline oral
squamous cell carcinoma
Biosensing of biofilm: detecting volatile organic compounds using a
dog’s nose
Environmental awareness: Recreating the natural tissue niche in vitro
for cancer research
Investigating the role of Delta-like ligand 1, a Notch signaling ligand,
in breast cancer metastasis
Temperature as a confounding variable in oncolytic virotherapy for
canine melanomas
In vitro effects of the chemotherapeutic agent water-soluble paclitaxel
on canine mast cell tumor cell lines
Why won’t you eat me? Investigating melanomas’ unique resistance to
phagocytosis through RNAi
The effects of natural and synthetic retinoid therapy on feline
squamous cell carcinoma cell lines
Characterization of exosome miR-9 expression in canine osteosarcoma
Transient Receptor Potential Melastatin 4 channel enhances
osteogenesis of rat dental pulp stem cells
Effects of caffeine on injured and uninjured intervertebral discs in a
whole organ culture model
In vivo evaluation of a novel HER2 inhibitor, lapatinib, in
osteosarcoma
A Novel Mechanism that Regulates the Surface Levels of NKp46 on
Natural Killer Cells
Effects of src homology 2 containing inositol 5-phosphatase (SHIP)
Inhibitors on canine osteosarcoma cells
Tumor Microenvironment Analysis via High Throughput Proteomics
after Poly(I:C)ZnO Treatment
Characterizing ischemic wound healing in jejunal epithelial cells in
porcine neonates and juveniles
Diallyl trisulfide inhibits epithelial cell adhesion molecule signaling in
hepatocellular cancer
Immortalization of Feline Mesenchymal Stem Cells with Feline
Telomerase and Novel Splice Variant Isoforms
Hyaluronan processing and function in the progression of breast cancer
Characterization of canine bone marrow mesenchymal stem cells
towards development of brain repair strategies
232
238
239
246
250
258
264
266
271
271
273
275
278
284
285
293
295
297
298
300
303
307
Developmental Biology
Banducci
Caitlin
20
Bucak
Carter
Emily
Samantha
46
57
Dellinger
Marion
88
Impact of Paternal and Maternal Preconception Stress on Offspring
Neurodevelopment and Stress Response
Biomechanical risk factors in the development of osteochondrosis in
Standardbred pacers and trotters
Do male sticklebacks use visual or olfactory cues to assess the history
of a potential mate?
60
100
113
119
134
Last Name
First Name Poster
#
Devereaux
Hayes
Joseph
Allison
90
136
Mertens
Micaela
246
Moorhead
Pruett
Wil
Katherine
254
292
Sato
Yu
318
Schenk
Soltis
Alex
Emily
323
353
Timonin
Mary
385
Abeywardena
Caceres
Emmitt
Shanes
Valeria
Nicole
2
50
100
Hanzel
Amanda
131
Lou
Jon
208
Manning
Hannah
223
Ondera
Caitlin
274
Pérez Irizarry
Cristian
279
Shearin
Abigail
334
Vijan
Stephanie
400
Watson
Claire
413
Abstract Title
Page
#
Perinatal growth hormone imprinting of hepatic cytochrome p450
expression in female rats
Monitoring bull behavior to determine characteristics of bull
dominance and the role in social ranking
Examining the relationship between affective state and social rank in
group-housed gestating sows
Behavioral Temperament of rhesus macaques (Macaca mulatta) and the
association with gut microbiota
The effect of prenatal alcohol exposure on respiratory regulation during
sleep in the early postnatal period
135
158
Endocrinology
Role of Neural Inflammatory Processes in Estradiol Enhancement of
Cocaine Self Administration in Female Rats
Effect of a Western-style or a high fat coconut oil diet on metabolism
and adipose tissue steroidogenesis
β-cell glucagon-like peptide-1 receptor improves islet morphology
after vertical sleeve gastrectomy in mice
Using behavior to establish a timed oxytocin protocol for estrous
suppression in mares
Licorice root: a phytoestrogen supplement with neuroprotective effects
on cognition
Effects of intravenous fluid therapy on serum osmolarity in
hospitalized dogs
Adipose AKT2 controls circulating free fatty acids and systemic insulin
resistance
Association of progestogens with inflammation and immunity in
critically ill foals
Deletion of liver fatty acid binding protein increases liver fat, bone
marrow fat and bone mineral density
213
217
236
249
252
267
283
91
115
140
156
194
202
227
230
257
290
297
Epidemiology/Public Health
Alexander
Derecka
5
Arroyo
Geraline
10
Bagnall
Hannah
15
Banuelos
Rosa
21
Beasley
Erin
26
Socioeconomic status of population and negligence of dogs with
visceral leishmaniasis in Araçatuba-SP, Brazil
Comparison of MALDI-TOF and PFGE for strain-typing
Staphylococcus aureus isolated from cow’s milk
An investigation into the epidemiology of B. odocoilei in Ontario ticks
and cervids
Prevalence and pathology of Trypanosoma cruzi in coyotes and
raccoons in a disease-endemic region of Texas
Pet carriage of Staphylococcus aureus and S. pseudintermedius in the
households of children with asthma
61
93
95
98
101
103
Last Name
First Name Poster
#
Birkner
Elise
35
Bishop
Black
Brianne
Kelley
36
38
Cashin
Kaitlyn
59
Charles
Jenna
67
Chen
Aolei
68
Croughan
Carolyn
82
Daniels
Demars
Barbara
Fanny
85
89
DiPastina
Ann
93
Dunfield
Engel
Elizabeth
Danielle
96
101
Gavron
Nancy
117
Ghearing
Hayter
Natasha
Sydney
119
137
Jones
Melissa
163
Jordan
Mary-ruth
165
Kramer
Caroline
180
Lantz
Maya
190
Li
Meng
197
Lucyshyn
Danica
210
Lyakhova
Tatyana
215
Mack
Tyler
216
Malcolm
Elizabeth
220
Martin
Alison
227
Mills
Kelly
250
Abstract Title
Page
#
Trypanosoma cruzi strain types infecting triatimone vectors and
primates at two research facilities in Texas
Development and performance analysis of a multiplex assay to detect
exposure to tick-borne diseases in dogs
Effect of low starch diet on health of sloth bears (Melursus ursinus) at
Cleveland Metroparks Zoo
Comparison of NIBP measurements obtained in a veterinary clinic
versus a home setting in hypertensive dogs
Systematic literature review and elicitation of expert opinion to assess
the 1997 FMD epidemic in Taiwan
Assessing mental health and identifying stressors among LSU School
of Veterinary Medicine students and alumni
Does in utero exposure to domoic acid, found in seafood, cause
epilepsy?
Optimizing antimicrobial use for mastitis to combat antimicrobial
resistance on US dairy farms
Make food great again: effect of protein source and amount on wholebody metabolism and weight gain
Refining approaches to assess microbiota and low-profile pathogens in
the tick vector, Amblyomma maculatum
Identifying antimicrobial resistance patterns in Salmonella enterica
Typhimurium from human and cattle sources
The Golden Retriever Lifetime Study: factors affecting owner
compliance after the first year of enrollment
Do you see what I see? Inter-rater reliability of a canine welfare
assessment used in dog breeding kennels
Rates and Risk Factors for Surgical Site Infections at the Ohio State
University Veterinary Medical Center
The effects of pet ownership on academic achievement among
veterinary students at St. George’s University
Management practices and disease perceptions among Minnesota
backyard flock owners following a HPAI outbreak
Sudden acquired retinal degeneration syndrome in Western Canada: a
retrospective (2002-2016)
Soil-borne protozoans enable persistence of facultative intracellular
pathogens in the environment
Immediate treatment of hyperketonemia ameliorates negative impacts
on milk production but not comorbidity risk
Radiographic and echocardiographic assessment of left atrial size in
dogs with mitral regurgitation
Adoption of recommended hand hygiene practices to protect public
health at agricultural fairs
Utility of ATP-bioluminescence measured by a luminometer to monitor
hygiene of milk replacer for dairy calves
108
62
108
109
120
124
124
131
133
135
137
138
141
149
150
159
172
173
180
185
189
195
198
198
200
204
215
Last Name
First Name Poster
#
Mitchell
Elspeth
252
Muller
Stefanie
258
Page
Lauren
276
Perry
Laura
281
Pike
Courtney
284
Rettkowski
Rice
Rebecca
Meredith
302
304
Smyth
Lauren
351
Sorg
Janna
356
Stabler
Emily
357
Stenger
Robert
359
StenkampStrahm
Stern
Chloe
360
Daniel
363
Storms
Suzanna
365
Strathe
Erin
367
Tarlowe
Stephanie
378
Thompson
Alexis
384
Trearchis
Deanna
388
Turinski
Tiffani
390
van Batavia
Ashley
393
Vincent
Emily
401
Vonderohe
Caitlin
402
Ware
William
412
Wright
Courtney
427
Abstract Title
Page
#
Antibiotic alternatives for the reduction of morbidity and mortality in
veal calves
Characterizing interactions between location, land use, and domestic
dog health in Panamanian communities
Efficacy of a new diagnostic procedure, the Mini-FLOTAC, in the
diagnosis of canine Giardia infection
Neutralization of chlorhexidine-containing products in a clinical hand
hygiene efficacy study
Venturing into the unknown: characterization of the aerobic cultivable
bacteria in wounds of wildlife species
Active veterinary surveillance of emerging Ixodes scapularis tick
populations in Michigan
A Comparison of Lyme disease Strain Diversity in Michigan Island
and Mainland Sites
The effect of more liberal milk allowance in the first week of life on
performance of preweaned dairy calves
Benefits of a therapeutic horseback riding program on veterans
suffering from PTSD or traumatic brain injury
Helminths in mountain gorillas: conservation through intestinal
parasite monitoring
Microbial community structures in E. coli O157 shedding and nonshedding dairy cattle; a preliminary analysis
Use of susceptible and avirulent E. coli to reduce antimicrobialresistant coliforms in milk-fed calves
The effects of human interactions on gastrointestinal parasites in
Mediterranean mouflon in southern France
A systematic review of the literature to identify and quantify host and
vector competence and abundance of JEV
Diagnostic accuracy of Keto-Test milk strips for cowside detection of
elevated milk β-hydroxybutyrate
Evaluation of refractometry to detect failed transfer of passive
immunity in pre-weaned beef calves
Prevalence of Failure of Passive Transfer, Dehydration, and Health
Outcomes in Veal calves on Day of Arrival
Characterization of the fecal microbiome from EHEC positive and
digital dermatitis negative beef cattle
Survey of infectious diseases in free-ranging Chilean Humboldt
penguins (Spheniscus humboldti)
Field research analysis of free-roaming cat populations on The Ohio
State University’s campus
The effect of antibiotic-free management on the environmental impact
of swine production
The effects of target distance and gun type on accuracy of 8cc darts
used in remote drug delivery systems
Detection of influenza A virus on inanimate surfaces at agricultural
fairs
216
63
219
228
231
232
241
242
266
268
269
270
270
272
273
274
279
282
284
285
287
291
291
296
304
Last Name
First Name Poster
#
Crombie
Kathryn
81
Falsey
Ferrero
Rachel
Nicole
103
106
Hammes
Joseph
129
Hanes
Steven
130
Healey
Eleni
138
Hosner
James
146
Howard
Hunter
Larrabee
Jennifer
Jacquelyn
Shannon
147
154
191
Ledesma
Eric
195
Maier
Kaitlin
219
Nilles
Jacob
267
Oldeschulte
James
270
Schade
Schrock
Seana
Kelly
321
327
Siegrist
Sirois
Melissa
Alexis
337
339
Smit
Michelle
342
Sorenson
Stilin
Juli
Allison
355
364
Stromberg
Stephanie
370
Vella
Gianna
397
Walker
Meghan
408
Wanner
Nicole
409
Whittmore
Jerrianne
419
Abstract Title
Genetics/Genomics
The genetics of familial hypertrophic cardiomyopathy in the Siberian
cat
Glutathione S-transferase-mu polymorphisms and cancer risk in boxers
Development of a qPCR assay for Altered Schaedler Flora colonized
mice utilizing the groEL gene
Reproductive performance applied to genetic merit and increasing
insemination rate of anovular Holsteins
Compromising X-linked gene silencing in mature B cells through
deletion of the long noncoding RNA Xist
Genome-wide association study seeks connection between stifle
morphology and cranial cruciate ligament disease
Use of the Collaborative Cross to Assay the Impact of Inter-Individual
Variability on TCDD-mediated Signaling
Discovery of a genetic mutation that causes sudden cardiac death in toy
Manchester terriers
Validation of putative chromosome 14 risk alleles for osteochondrosis
in Standardbred horses
Investigation of sarcolipin as a cause of recurrent exertional
rhabdomyolysis in horses
Discovery of common genetic risk loci for canine cruciate ligament
rupture by validation GWAS
Development of large animal models for a compound heterozygous
human disease using CRISPR/Cas9
Molecular etiology of intrahepatic portosystemic shunts in the Nova
Scotia Duck Tolling Retriever
Examination of genetic variation in CD18 of cattle as a marker for
resistance to respiratory disease
Genetic investigations of the AMH and AMHR2 genes in canine
persistent Mullerian duct syndrome
Discovery of the causal variant for acquired laryngeal paralysis
polyneuropathy in the dog
Genotype and phenotype of sudden acquired retinal degeneration
syndrome (SARDS) in dachshunds
Using human cell-specific data and correlation to cell proportion to
predict pig neutrophil-specific genes
Development of a next-generation sequencing method for veterinary
forensics
Impact of a paternally inherited allele for polysaccharide storage
myopathy on muscle pain and myodegeneration
Changes in the equine gut microbiome associated with hospital stay
64
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131
142
143
155
155
159
163
164
167
186
188
200
224
225
251
254
259
260
261
268
272
275
289
294
295
300
Last Name
First Name Poster
#
Wong
Stephanie
426
Zuber
Emily
436
Abstract Title
Page
#
Deleterious glutathione-S-transferase-pi (GSTP1) polymorphisms may
contribute towards Boxer dog lymphoma risk
Placental abnormalities and fetal growth restriction as a consequence
of preconception male alcohol exposure
303
Mast cells in allergies and in asthma- does particulate matter (PM)
activate mast cells?
Evaluation of health in Texas tortoises via multiple patient-side
analytes
Effects of microaggregate filters on canine erythrocyte viability and
morphology
91
99
168
183
242
Characterizing immune regulation of canine heartworm Dirofilaria
immitis infection in Aedes aegypti mosquitoes
Surveillance of Florida snails for Parelaphostrongylus andersoni and
Parelaphostrongylus tenuis
Assessing key proinflammatory cytokines in equine peripheral blood
mononuclear and monocytic cells, in vitro
Investigating IgG subtype responses in horses infected with different
mutations of EHV-1
The Effect of High Density Lipoproteins on Bovine Neutrophil
Activation
Pharmacodynamic assessment of a panel of immunosuppressant drugs
on ex-vivo canine T-lymphocyte proliferation
Optimizing the broth environment for large volume growth of
“Brachyspira hampsonii”
Evaluation of Plasmodium knowlesi in vitro susceptibility to
methotrexate
Linking canine distemper virus susceptibility to SLAM sequences in
wildlife species
Generation of a recombinant PIV5 virus expressing Zika antigen for
vaccine development
In vitro evaluation of cell mediated immune responses to respiratory
antigens in high and low risk beef calves
The proline utilization system of Brucella abortus is required for
virulent infection
Development and validation of a PCR platform for the sensitive and
specific detection of Bartonella rochalimae
92
308
Hematology
Abdel Massih
Baley
Ienello
Lake
Rice
Cherein
Susan
Lauren
Bathilda
Deja
1
17
156
186
303
Rodenbaugh
Taylor
Cassandra
Michelle
308
382
Williams
Jessica
424
244
281
302
Infectious Disease and Immunology
Aguiar
Ariel
4
Allgood
Hillary
7
Asklof
Aust
Barnett
Kendra
Rebecca
Brian
11
13
23
Baumgardner
Rachel
25
Bickers
Bridget
32
Bishop
Kaitlin
37
Boire
Francois
41
Briggs
Brockhurst
Cassidy
Jacqueline
44
45
Cahill
Bridgette
51
Callahan
Robert
52
Cash
Kaycee
58
Castell
Caudill
Natalie
Mitchell
61
62
Chan
Dennis
63
65
94
96
97
102
103
106
109
111
112
113
116
116
119
121
121
122
Last Name
First Name Poster
#
Chandler
A’Jah
65
Chang
Kevin
66
Clark
Megan
71
Clark
William
72
Cluett
Matthew
75
Coleman
Denver
76
Crilly
Nathan
80
Daugherty
Dickson
Elizabeth
Rachel
86
91
Draper
Janna
94
Fedderly
Folkerts
Galya
Stephanie
105
109
Fousse
Samantha
111
Frady
Kristine
112
Gilfeather
Glick
Christa
Melissa
120
122
Gustafson
Kevin
124
Hagstrom
Melena
126
Hardy
Hausmann
Jennifer
Katharine
132
135
Hecker
Kari
139
Henson
Christopher
142
Horng
Katti
145
Icaza
Melissa
155
Jeffreys
Roxanna
158
Jimenez
Monica
159
Abstract Title
Page
#
Characterizing the mechanisms of action of novel chemical inhibitors
of Mycobacterium smegmatis growth
Fecal toxin cytotoxicity as a measure of clinical disease in mice
infected with Clostridium difficile R20291
Characterization and optimization of ex vivo T cell expansion for redirected T cell therapy
Early effects of ivermectin on intraperitoneal Brugia pahangi infection
in Meriones unguiculatus
Effects of IL-10 modulation on nerve pathology in C. jejuni induced
Guillain-Barre mouse models
Effect of temperature on Veronaea botryosa growth and cytotoxicity to
Acipenser transmontanus skin cell lines
TAM Receptor Expression Is Altered During Zika Virus Infection in
Pregnant Mice
Evaluating the effect of Fortiflora probiotic therapy on feline
Tritrichomonas foetus infection
Microneutralization assay: an economical method for determining
vaccine recommendations for swine influenza
Isolation of antibiotic-producing bacteria from New River Valley
(NRV) soil
Does re-emerging St. Louis encephalitis virus in California show
increased fitness compared to older strains?
Characterization of the pathogenic potential among different species in
the spotted rickettsial group
Estrogen protects progestin-treated mice against intravaginal
transmission of cell-associated HIV-1
The use of a B cell tetramer to evaluate the memory immune response
to PRRSv
Vertical non-transovarial transmission of Rickettsia felis in the cat flea,
Ctenocephalides felis
Examination of Pseudoloma neurophilia infection in the central
nervous system of zebrafish (Danio rerio)
Commercialization of an ELISA for Ornithobacterium rhinotracheale
using hemolytic and non-hemolytic isolates
Role of the gerQ determinant in exosporium structure in Bacillus
anthracis
Beneficial effects of probiotic microbes on inflamed gut mucosa in the
rhesus macaque model of HIV infection
Degradation of complement in the presence of Schistosoma mansoni in
human serum
Characterization of the skin microbiome of dogs with strong body odor
and the effect of a spot-on product
Impact of Helicobacter pylori on the infant lung: airway microbiome
and immunomodulation of airway epithelium
123
66
123
126
126
128
128
130
133
136
137
143
145
146
146
150
151
152
153
156
158
160
161
163
168
169
170
Last Name
First Name Poster
#
Johnson
Catherine
160
Johnson
Lauren
161
Jong
Khambholja
Ruby
Chantelle
164
168
Kim
Sohyun
171
Kirby
Klein
Ashley
Hannah
172
173
Knebel
Emily
175
Kobluk
Kristi
176
Kuball
Kristin
182
Lamb
Holly
187
Landis
Casey
188
Lawnichak
Tyler
194
Lee
Amber
196
Lin
Dian Dian
199
Linn
Sarah
202
Livengood
Kristen
203
Loeschel
Courtney
204
Long
Mackenzie
205
Lundquist
Maeroff
Matthew
Jacqueline
213
218
Mangosing
Sara
221
Mason
Caitlin
230
Mattia
Michael
232
May
Emily
233
Abstract Title
Page
#
MARCKS protein as a therapeutic target in the treatment of equine
asthma
Susceptibility of equine monocyte subsets to equine herpes virus type I
infection
Using in-lab evolution to explain how protozoan parasite Toxoplasma
gondii is such an exceptional generalist
Isolation and cloning of feline Macrophage Colony Stimulating Factor
for optimizing macrophage differentiation
Dysbiosis and bile acid dysmetabolism in dogs with steroid responsive
enteropathy
Urine for a surprise: Evaluating the use of expanded quantitative urine
culture on feline samples
Genetic knockout screens identify host genes required for zika virus
replication
Using Cold Atmospheric Plasma as an Alternative Treatment for
Chronic, Antibiotic Resistant Wounds
Evaluation of the roles of plasmin and CCL20 on nerve damage in an
inflammatory M. leprae infection model
Characterization of a co-culture model for observation of Francisella
tularensis infection
Influence of donepezil on the development of Toxoplasma gondii and
its potential relationship to Alzheimer’s
Examination of insectivorous bats (Molossus molossus) in Grenada for
trematodes and Neorickettsia DNA
Two methods of Clostridium difficile toxin purification for detection by
MALDI-TOF MS
Experimental modeling of the nonspecific protective effects with
measles virus vaccination
Biofilm formation by mastitis-producing Staphylococcus aureus and
inhibition by 2-aminoimidazole compounds
Evaluation of methods to preserve field samples for detecting elephant
endotheliotropic herpesviruses
Induced GI tract microbial dysbiosis does not accelerate disease
progression in SIV-infected Asian macaques
Comparison of lymphocyte count and proviral load in beef and dairy
cattle infected with bovine leukemia virus
Generation of Infectious Bronchitis Virus (IBV) M41 cDNA clones for
use in chimeric IBV virus construction
Native and recombinant PD-1, PD-L1, and PD-L2 variants for study of
porcine circovirus associated disease
Fate of exosporium proteins BclA, BclB, and BxpB in germinating
Bacillus anthracis spores
Antigen presenting cells promote protective immunity in catfish
exposed to live Edwardsiella ictaluri vaccines
170
67
171
172
174
176
176
177
178
178
181
184
184
187
188
190
191
192
192
193
197
199
201
205
206
207
Last Name
First Name Poster
#
Mendoza
Gerardo
244
Mercer
Middlebrooks
Mosca
Stephen
Sarah
Leandra
245
247
255
Moyer
Murray
Elizabeth
Olivia
257
259
Myers
Laurel
261
Niel
Kayla
265
Olney
Perier
Erika
Nadege
271
280
Phillips
Elsie
282
Poole
Porter
Tyler
Stephanie
287
288
Powers
PradoSanchez
Price
Jonathan
Evymarie
289
290
Emma
291
Putman
Richardson
Rimmelin
Ashley
Ella
Alesha
294
305
306
Rose
Hailey
309
Rowe
John
314
Russell
Sablotny
Schacher
Scheffe
Scott
Ellen
Elizabeth
Michael
Gretchen
Jenna
315
316
320
322
331
Siddons
Giles
336
Smith
Courtney
344
Smith
Katelin
345
Abstract Title
Page
#
A single mutation in the vaccinia virus RNA polymerase inhibits
multiple host restriction factors
Domains of CD163 involved in infection of Type I and Type II PRRSV
Comparative immunogenicity of attenuated Salmonella vaccine strains
dependent on parental origin and genotype
Attempt to confirm absence of Avian Bornavirus in a previously
shedding flock of cockatiels (N. hollandicus)
Characterizing infection of Jamaican fruit bats with bat-derived
influenza-like viruses, HL17N10 and HL18N11
The growing feather as a model for novel evaluations of Marek’s
disease genetic resistance
The impact of mastitis on maternal behavior in dairy cows: a pilot
study
Role of Mouse Sca1+ Lung Mesenchymal Stem Cells in Bacterial
Pneumonia
Environmental Reservoirs of Francisella tularensis: Mechanisms for
Transmission of Tularemia
Nickel homeostasis and bacterial pathogenesis: Examining the NikR
regulatory system in Brucella abortus
Characterization of the role of type I and type III interferons in acute
and chronic disease
Disparate virulence in two plaque variants of Theiler’s murine
encephalitis virus, DA strain
Mutant XYZ is associated with hyper-activation of mTOR and T cell
senescence
Intestinal epithelial cells NF-kB regulates host response to ingestion of
low doses of cadmium
Effect of litter type on survival of Tritrichomonas foetus in feline feces
Effects of deworming and vaccination on seroresponse to BVDV1 and
BHV1 in stocker calves
Schistosomes recruit host regulatory proteins to resist complement
attack
Comparison of molecular methods in the characterization of
Malassezia on the skin of healthy and allergic dogs
Eosinophil activity is not altered by respiratory syncytial virus
infection in the cotton rat
212
68
213
214
218
219
220
221
223
226
230
231
234
234
235
235
236
237
243
243
245
247
248
248
250
251
256
258
262
263
Last Name
First Name Poster
#
Smith
Katherine
346
Smith
Richard
349
Stelly
Philip
358
Strumpf
Tageant
Alyssa
Connor
371
374
Tang-Wing
Cassandra
377
Tatro
Melissa
379
Taupier
Rachel
380
Tavella
Vincent
381
Tenborg
Elizabeth
383
Varley
Ashley
395
Velazquez
Eric
396
Vermillion
Meghan
398
Wahl
Alexandra
404
Walker
Anna
406
Williams
Courtney
423
Wronski
Yang
Sarah
Stephanie
428
429
Young
Kimberly
431
Zabrecky
Kristin
432
Zlotnick
Marta
434
Zorn
Chelsea
435
Zygelyte
Emilija
437
Adams
Daniel
3
Ballard
Casper
Kristen
Daniele
18
60
Abstract Title
Page
#
Bacterial killing ability of vampire bats and associations with
individual body condition and feeding ecology
VP22 transport from differentiated Neuro2A cells in microfluidic
chambers to epithelial cells and vice versa
γδ T Cells play a critical role in the inflammatory response to
Paramyxovirus infection in mice
Vaccination of small ruminants against Haemonchus contortus using
fecal antigens
Aedes Aegypti mosquito transmission and the development of STAT2
KO hamster animal model for Zika virus
Marek’s disease infectivity in genetically susceptible and resistant
chickens
Effects of Bedoukian compound concentrations on gastrointestinal
nematode development in sheep
Control of lupus nephritis by manipulating gut microbiota during
active disease
How does Toxoplasma gondii deliver effector proteins that control host
cell functions
Comparison of virus isolation from fabric for use in herd surveillance
of viral bovine respiratory disease
Commensal Enterobacteriaceae Drive Variable Salmonella
Colonization Resistance of Mice from Different Vendors
Estriol protects female mice against influenza A virus by reducing
pulmonary inflammation
Florfenicol resistance in chloramphenicol-resistant canine isolates of
Staphylococcus pseudintermedius
Assessment of gastrointestinal nematode parasitism in dairy calves
housed in calf hutches
Identification of Aeromonas hydrophila virulence genes providing
complement resistance
Host-oriented inhibitors of Lassa fever virus budding
Prenatal infection with Listeria monocytogenes and risk for
neurodevelopmental defects in pregnant mouse model
Phenotypic evaluation of multicopy single-stranded DNA mutants to
determine functional regions
Gut microbiota analysis of an Alzheimer’s Disease transgenic rat
model
Comparison of infectivity of Zika virus strains in primary adult murine
neurons
Isolation of uniquely recognized salivary gland antigens to interfere
with feeding performance of ixodid ticks
Demethylase inhibitor reduces equine herpesvirus-1 expression in vitro
263
Molecular Biology and Biochemistry
Determining the effects of SMAC mimetic drugs on vascular
endothelial cells
69
265
269
276
277
279
280
280
281
282
288
288
289
292
293
302
304
305
306
306
307
308
309
92
99
120
Last Name
First Name Poster
#
Chung
Sarah
70
Clarke
Samantha
73
De La
Espriella
Duliepre
Jose
87
95
Fasig
StephieAnne
Joseph
104
Guan
Chiyu
123
Haiderer
Elizabeth
127
Hendrix
Madeleine
141
Hristova
Koenig
Teodora
Kelsey
148
177
Kraneburg
Carol
181
Laurence
Hannah
192
Mann
Olivia
222
Marinoff
Jacqueline
225
Mitchell
Freelie
253
Nagel
Pador
Pires
Reinhard
Jonathan
Sean
Elena
Natalie
262
275
286
300
Reising
Abby
301
Schoenberger
Jenna
326
Schulte
Rachael
328
Schulze
Laura
329
Seelman
Amanda
332
Swanson
Tracy
373
Abstract Title
Page
#
Effect of FABP1/SCP-2/SCP-X Gene Ablation (TKO) on the
Endocannabinoid System in Mice
Alterations in myofilament-associated signaling complexes in dogs
with naturally occurring DCM
Purification and stability screens of conserved virulence factor A in
Streptococcus pyogenes
125
Evaluating body condition index with DEXA and deuterium oxide in
big brown bats (Eptesicus fuscus)
Expression and Purification of PrM and E Proteins of Zika Virus in Sf9
Cells
Characterization of the spectrum of activity and mechanism of a pHselective inhibitor of M. tuberculosis
Identification and characterization of Yersinia ruckeri from farm-raised
catfish in Mississippi
The effect of oral sodium bicarbonate “milkshake” on serum total
carbon dioxide concentration (TCO2) in horses
Contraception of feral cats with phage-based vaccines targeting
gonadotropin releasing hormone (GnRH)
The architecture of the Flavivirus 3’ untranslated region, pathogenic
RNA production, and viral cytopathicity
Characterization of N-linked glycan profiles in bat gastrointestinal
tissues specific to influenza A viruses
Optimization of extraction and quantitation of cfDNA from equine
plasma
Optimization of DNA Vaccine Targeting GnRH Receptor for Animal
Contraception
Structural modifications to K+ channel, Kv1.1, in a wild rodent
resistant to scorpion venom
Demonstrating C. botulinum neurotoxin heavy chain cytosolic
localization via modified Kirby-Bauer assays
Effects of calorie restriction on radiation resistance of reserve intestinal
stem cells
Paraventricular nucleus neurons target nucleus tractus solitarii via CRH
receptors and oxytocin in hypoxia
Down stream effects of bovine respiratory disease complex on
myocardial gene expression
Identification of Arginine Vasopressin producing neurons in the
amygdala of domestic cats
Defining the relationship between female sex hormones and
myocardial fibrosis in aortic-banded mini-swine
70
127
134
138
142
152
154
161
164
179
181
186
201
203
217
221
228
233
240
241
253
254
255
256
277
Last Name
First Name Poster
#
Touitou
Florian
386
Voss
Brittany
403
Baker
Anna
16
Bancroft
Channing
19
Belter
Rebecca
29
Bentz
Kelsey
31
Blank
Carolyn
39
Burns
Jordan
47
Cabral
Troy
49
Chan
Jennifer
64
Daney
Jolani
84
Dionne
Madeleine
92
Eagalle
Thisuri
97
Fisk
Elizabeth
107
Ford
Jordan
110
Hsu
Steven
149
Janse van
Rensburg
Johnson
Claire
157
Tyler
162
Kluz
Michael
174
Kustasz
Lauren
185
Landreth
Victoria
189
Lin
Megan
200
Lind
Lori
201
Lovering
Ludwig
Samantha
Latasha
209
212
Abstract Title
Page
#
Development of less invasive functional assessments of malignant
hyperthermia (MH) genotypes
Purification of hantavirus cap snatching endonuclease
283
Volumetric analysis of the hippocampus and amygdala in California
sea ions with domoic acid induced epilepsy
Effects of 17-α-estradiol treatment on stifle joint morphology in aging
mice
Comparison of enterotomy leak pressure among fresh, cooled, and
frozen/thawed swine jejunal segments
Susceptibility of Amblyomma americanum, Dermacentor variabilis
and Rhipicephalus sanguineus to acaricides
Variation in bronchial size during respiration and cough in healthy
compared to malacic dogs
Identification of biomarkers for xenoestrogenic exposure using a
prepubertal porcine model
Improving adjunctive treatment of drug resistant diabetic foot
infections
Fluoroscopic determination of thoracic dimensional changes in healthy
dogs
The prophalactic effect of acetylsalicylic acid on astrogliosis in a
mouse model of scrapie
Evaluating the role of female sex hormones on left ventricular
mechanics in aortic-banded mini-swine
Determination of causes of mortality in captive psittacines submitted
to the Ontario Veterinary College
Comparison of Contemporary Aqueous and Alcohol-Based Scrub
Agents for Rodent Surgical Skin Preparation
Parvovirus detection by PCR and ISH is associated with myocarditis
and cardiomyopathy in young dogs
Construction and expression of full-length canine circovirus molecular
clone
Do honey bee colonies (Apis mellifera) prefer feed contaminated with
neonicotinoids?
Coagulopathy in Crotalus viridis envenomation, attenuation by carbon
monoxide releasing molecule - 2 in vitro
Evaluation of vertebral articular process dysplasia and the
thoracolumbar myelopathies in pug dogs
Optimization of ultrasound settings for better determination of cystolith
size in vitro
Exploration of tongue muscle pathology in canine degenerative
myelopathy
Progression of vertebral bone pathology in mucopolysaccharidosis VII
dogs from birth to skeletal maturity
The effects of tongue injection of CTB-SAP on ventral hypoglossal
motor neurons: a novel model of ALS
98
Pathology
71
292
100
105
106
110
114
115
122
132
136
139
144
145
165
169
171
177
183
185
190
191
195
196
Last Name
First Name Poster
#
McGeehan
Megan
235
McLean
Victoria
236
McSweeny
Melvin
Ian
Rachel
239
243
Moses
Amber
256
Mustonen
Allison
260
Oelfke
Thomas
268
Rose
Tierra
310
Schuster
Chelsea
330
Sirochman
Anna
338
Slabaugh
Taylor
340
Streu
Shayna
368
Vadala
Clara
392
Walker
Ashley
407
Werner
Melissa
416
Weyna
Alisia
417
Whisenant
Katrijn
418
Abstract Title
Page
#
Characterization of tertiary lymphoid organs in a natural model of
Multiple Sclerosis
Morphology of peritoneal endometriotic lesions using an in-vivo
mouse model of retrograde menstruation
Oviduct morphology and Prolactin receptor expression in N. American
River Otters and Asian Small Clawed Otters
Changes in periarticular bone secondary to osteoarthritis in
chikungunya-infected and naive mice
How to smoke your bacon: An investigation on the effects of
environmental inhalants in a porcine model
A proposed method for functional quantitation of carpal joint laxity in
canine mucopolysaccharidosis type I
Loss of parvalbumin-immunoreactive interneurons in epileptic
California sea lions
Intraperitoneal ethanol injection for humane euthanasia in zebra finches
(Taeniopygia guttata): a pilot study
Is there evidence for canalithiasis or cupulolithiasis as an etiology of
canine idiopathic vestibular disease?
Frequency and complexity of arrhythmias in rhesus macaques with and
without hypertrophic cardiomyopathy
A questionnaire-based study on predisposing and risk factors for gastric
dilatation volvulus in dogs
Carnoy’s vs. Formalin: a comparison of two fixatives in colonic
samples from weaned piglets
Ambulatory ECG, heart rate variability and cardiac stress in cats with
subclinical hypertrophic cardiomyopathy
Accessing Potential Causes of Repeat Breeder Dairy Cows through
Cytology, Bacteriology, and Histopathology
Feline ocular and periocular squamous cell carcinoma: a retrospective
study of 374 cases
Targeting glycolysis reduces inflammation in a murine models of
rheumatoid arthritis and Crohn’s disease
208
208
210
212
218
220
224
245
255
259
260
274
286
294
298
299
299
Pharmacology/Toxicology
Alexander
Kayla
6
AvalosCavero
Barbar
Barbara
14
Megan
22
Bennett
Joshua
30
Binns
Jamie
33
Brado
Glenn
43
Cardwell
Jason
54
Flow cytometric evaluation of canine lymphocytes after oral
mycophenolate mofetil
Study of human and mouse lung cells responses to carbon nanotubes to
assess their pulmonary disease potential
Evaluation of improvement in joint function from intra-articular
injection of NV-08 in dogs with OA joint pain
Acute toxicity evaluation in rats of potential chemical warfare agent
antidotes
Sympathetic neurotransmitter induced vasoconstrictions in high fat and
control diet fed rats
Effect of intranasal cannabinoid administration on an experimentally
induced rhinosinusitis mouse model
Plasmin induces up regulation of matrix metalloproteinase-13 in
murine bone marrow derived macrophages
72
93
97
101
105
107
112
117
Last Name
First Name Poster
#
Carothers
Currie
56
Chu
Colin
69
Ebelle
Danielle
98
Esposito
Gena
102
Furst
Emmanuelle
113
Furst
Nicole
114
Galang
Kristopher
115
Holdridge
Julie
144
KelleyCollier
Khan
Koestel
Parrish
167
Michael
Zoe
169
178
Mariampillai
Anujah
224
Masters
Allison
231
Melhado
Ashlee
241
Miller
Travis
248
Nietlisbach
Nicole
266
Oltman
Wesley
272
Onaga
Robert
273
Pulczinski
Jairus
293
Ramirez
Berrios
Randall
Karen
295
Chelsea
297
Roseman
Brandi
311
Rowbotham
Nicole
313
SanabriaOjeda
Laura
317
Abstract Title
Page
#
Respiratory responses to short term exposure to second hand smoke in
a Guinea pig asthma model
In vitro endothelial cell wound healing in response to red wine
polyphenols resveratrol and quercetin
The Effect of Sandstorm Particles on Antigen-Presenting Dendritic
Cells
Recreating geophagy in a captive psittacine species, Myiopsitta
monachus
Primary neuron culture autapses as a model for investigating opioid
receptor desensitization
Cardiovascular effects of epinephrine on Alligator mississippiensis
recovering from isoflurane anesthesia
Alterations in hepatic glucose metabolism following direct exposure to
organochlorine pesticide metabolites
The effect of chronic Bisphenol A exposure on estrogen-sensitive gene
expression (ER α & ER β, ERR γ)
Is dietary exposure to bisphenol A causing phenotypic changes in
dogs?
Biomonitoring the effects of BPA and its metabolite BPAG during fetal
life
Effects of anti-inflammatory glucocorticoids on hemodynamics and
cardiac function in clinically healthy dogs
Pathophysiologic environmental factors diminished efficacy of
ceftiofur and penicillin with gentamicin
Effect of a supplement containing curcumin on lameness due to
osteoarthritis or degenerative disease
Thermal nociception and gastrointestinal function following
administration of Simbadol in rats
Seasonal variation of 25-hydroxy-vitamin D in two captive Eastern
black rhinoceros (Diceros bicornis michaeli)
Maternal markers of inflammation and oxidative stress following air
pollution exposure during pregnancy
118
Evaluating body condition score with body weight on serum drug
levels of phenobarbital and potassium bromide
Parkinson disease modeling in parkin-deficient mice: testing synthetic
mitochondrial-targeted antioxidants
Involvement of increased CB1R activation in altered social play
induced by developmental chlorpyrifos exposure
Effects of the JAK-inhibitor AZD1480 on activated dorsal root ganglia
and itch behavior in mice
73
125
139
141
147
147
148
162
174
175
179
202
206
211
214
223
226
227
237
238
239
246
247
249
Last Name
First Name Poster
#
Schettler
Michael
324
Senft
Tyler
333
Slimack
Katie
341
Smith
Kayla
347
Smith
Tamas
Samuel
David
350
375
Turner
Zachary
391
Vetter
Nicholas
399
Ward
Kinnon
411
Wilcox
Parker
422
Abstract Title
Page
#
No evidence for sexual dimorphism of nitric-oxide (NO) mediated
vascular control in rat skeletal muscle
Two trials to investigate techniques for canine Vitamin D
supplementation
The role of lidocaine and meloxicam in pain management for bull
castrations
Tricaine methane sulfonate (MS-222) anesthesia in fishes: Relationship
to thermal divergence
The effects of inflammation on the pharmacokinetics of flunixin in
steers using in vivo ultrafiltration
Fescue toxicosis in grazing beef cattle: impact of environmental
temperature and humidity
Rodent Models of Ozone-Induced Eosinophilic Rhinitis: Species- and
Strain-Dependent Variations
The effects of ischemic stroke on cardiac inflammation in
normotensive rats
Efficacy and safety of topical nitric oxide donors on decreasing
intraocular pressure in healthy canine eyes
252
74
257
261
264
265
278
286
290
296
301
Symposium Participants by
Vet School
Last Name
First Name
Poster
#
Atlantic Veterinary College
Auburn University CVM
Colorado State University CVM
Cornell University CVM
Cummings SVM at Tufts University
Iowa State University CVM
Perry
Kraneburg
Mitchell
Roberson
Cabral
Cutcliffe
Furst
Hunter
Johnson
Jong
Kumar
Lyakhova
Millman
Pannone
Pires
Porter
Stenkamp-Strahm
van den Hoogen
Arica
Bickers
Calle
Duliepre
Ford
Healey
Herrold
Johnson
Lieberman
Lou
Lovering
Onaga
Tarlowe
Walden
Yang
Zygelyte
Icaza
Nguyen
Siddons
De La Espriella
Ferrero
Laura
Carol
Freelie
Rachel
Troy
Ross
Emmanuelle
Jacquelyn
Tyler
Ruby
Anisha
Tatyana
Zachary
Stephen
Elena
Stephanie
Chloe
Marjan
Andrea
Bridget
Jasmine
Stephie-Anne
Jordan
Eleni
Emily
Lauren
Seth
Jon
Samantha
Robert
Stephanie
Katherine
Steven
Emilija
Melissa
Chrystal
Giles
Jose
Nicole
281
181
253
307
49
83
113
154
162
164
184
215
249
277
286
288
360
394
9
32
53
95
110
138
143
161
198
208
209
273
378
405
430
437
155
263
336
87
106
75
Vet School
Last Name
First Name
Poster
#
Kansas State University CVM
Lincoln Memorial University CVM
Louisiana State University SVM
Hagstrom
Kobluk
Lundquist
Masters
Mertens
Oelfke
Oltman
Vella
Wilcox
Zhou
Fleming
Guan
Luu
Madden
Mercer
Pérez Irizarry
Schettler
Strathe
Warcholek
Zuber
Rice
Alexander
Aust
Ballard
Blank
Bollman
Carothers
Charles
Chu
Cleveland
Courville
Croughan
Frady
Hargis
Hughes
Koenig
Lamb
Moses
Phillips
Ramirez Berrios
Rash
Smith
Smith
Stelly
Melena
Kristi
Matthew
Allison
Micaela
Thomas
Wesley
Gianna
Parker
Xueying
Payge
Chiyu
Savannah
Kelsey
Stephen
Cristian
Michael
Erin
Stanislaw
Emily
Meredith
Derecka
Rebecca
Kristen
Carolyn
Mary Kate
Currie
Jenna
Colin
Ashley
Emily
Carolyn
Kristine
Jessie
Peyton
Kelsey
Holly
Amber
Elsie
Karen
Morgan
Kayla
Richard
Philip
126
176
213
231
246
268
272
397
422
433
108
123
214
217
245
279
324
367
410
436
304
5
13
18
39
42
56
67
69
74
78
82
112
133
152
177
187
256
282
295
299
347
349
358
76
Vet School
Last Name
First Name
Poster
#
Michigan State University CVM
Michigan State University Undergraduate
Michigan State University Undergraduate
Michigan State University- undergraduate
Midwestern University CVM
Midwestern University CVM
Mississippi State University CVM
Stovall
Tageant
Taupier
Baumgardner
Binns
Brado
Cardwell
Cluett
Daniels
DiPastina
Fisk
Haiderer
Hosner
Hughes
Kelley-Collier
Kim
Kluz
Kustasz
Maeroff
Maier
Melvin
Niel
Piegols
Putman
Reinhard
Schade
Schenk
Smyth
Stern
Streu
Tatro
Vetter
Ward
Yang
Chandler
Ebelle
Rice
Olney
Rose
Alexander
Bennett
Bucak
Gavron
Gilfeather
Kelsie
Connor
Rachel
Rachel
Jamie
Glenn
Jason
Matthew
Barbara
Ann
Elizabeth
Elizabeth
James
Courtney
Parrish
Eunbee
Michael
Lauren
Jacqueline
Kaitlin
Rachel
Kayla
Hunter
Ashley
Natalie
Seana
Alex
Lauren
Daniel
Shayna
Melissa
Nicholas
Kinnon
Stephanie
A’Jah
Danielle
Deja
Erika
Hailey
Kayla
Joshua
Emily
Nancy
Christa
366
374
380
25
33
43
54
75
85
93
107
127
146
151
167
170
174
185
218
219
243
265
283
294
300
321
323
351
363
368
379
399
411
429
65
98
303
271
309
6
30
46
117
120
77
Vet School
Last Name
First Name
Poster
#
North Carolina State University CVM
Hayter
Hendrix
Holdridge
Mann
May
McMullin
Middlebrooks
Moorhead
Moyer
Rowbotham
Stenger
Thompson
Varley
Walker
Ware
Williams
Anderson
Avalos-Cavero
Barbar
Beasley
Becher
Chang
Folkerts
Gavitt
Harris
Johnson
Joyner
Kirby
Kucera
LaVine
Livengood
Lopez
Medland
Melhado
Sablotny
Sanabria-Ojeda
Scheffe
Scott
Si
Sommer
Tamas
Walker
Wegner-Clemens
Young
Sydney
Madeleine
Julie
Olivia
Emily
Alan
Sarah
Wil
Elizabeth
Nicole
Robert
Alexis
Ashley
Anna
William
Jessica
Jessica
Barbara
Megan
Erin
Jessica
Kevin
Stephanie
Ashley
Elizabeth
Catherine
Jalika
Ashley
Cecilia
Danielle
Kristen
Ariana
Julia
Ashlee
Elizabeth
Laura
Gretchen
Jenna
Catherine
Samantha
David
Meghan
Erik
Kimberly
137
141
144
222
233
238
247
254
257
313
359
384
395
406
412
424
8
14
22
26
27
66
109
116
134
160
166
172
183
193
203
206
240
241
316
317
322
331
335
354
375
408
414
431
78
Vet School
Last Name
First Name
Poster
#
Ohio State University CVM
Oklahoma State University CVM
Ontario Veterinary College
Purdue University CVM
Bartholomew
Cashin
Casper
Galang
Geisler
Ghearing
Glick
Kramer
Linn
Manning
Martin
Mironovich
Mitchell
Pinnell
Rowe
Scheuermann
Smith
Stephenson
Trearchis
Vijan
Vincent
Werner
Wickware
Wright
Zabrecky
Bentz
Hardy
Landis
Rodenbaugh
Sirois
Bagnall
Clarke
Eagalle
Ludwig
Saturno
Wong
Bloom
Byerley
Devereaux
Draper
Haffner
Jones
Jordan
Lin
Kristen
Kaitlyn
Daniele
Kristopher
Jennifer
Natasha
Melissa
Caroline
Sarah
Hannah
Alison
Melanie
Elspeth
Erin
John
Logan
Katelin
Olivia
Deanna
Stephanie
Emily
Melissa
Taylor
Courtney
Kristin
Kelsey
Jennifer
Casey
Cassandra
Alexis
Hannah
Samantha
Thisuri
Latasha
Julia
Stephanie
Christopher
Sydney
Joseph
Janna
Abigail (Abbie)
Melissa
Mary-ruth
Dian Dian
24
59
60
115
118
119
122
180
202
223
227
251
252
285
314
325
345
362
388
400
401
416
420
427
432
31
132
188
308
339
15
73
97
212
319
426
40
48
90
94
125
163
165
199
79
Vet School
Last Name
First Name
Poster
#
St Georges University SVM
Texas A&M University CVM
The University of Queensland
Tuskegee University SVM
University of Calgary
University of California - Davis SVM
Mustonen
Rasche
Rose
Smith
Strumpf
Vonderohe
Weigel
Lantz
Lee
Tang-Wing
Banuelos
Birkner
Castell
Chung
Esposito
Hanzel
Jeffreys
Klein
Loeschel
Martinez
Murray
Oldeschulte
Pulczinski
Rimmelin
Schulze
Smith
Taylor
Tran
Vadala
Wahl
Watson
Cahill
Hunt
Lopez
Miller
Roseman
Williams
Schuster
Chan
Coleman
Fousse
Horng
Hsu
Laurence
Allison
Brittany
Tierra
Samuel
Alyssa
Caitlin
Ellen
Maya
Amber
Cassandra
Rosa
Elise
Natalie
Sarah
Gena
Amanda
Roxanna
Hannah
Courtney
Samantha
Olivia
James
Jairus
Alesha
Laura
Courtney
Michelle
Kim
Clara
Alexandra
Claire
Bridgette
Janna
Melissa
Travis
Brandi
Courtney
Chelsea
Jennifer
Denver
Samantha
Katti
Steven
Hannah
260
298
310
350
371
402
415
190
196
377
21
35
61
70
102
131
158
173
204
229
259
270
293
306
329
344
382
387
392
404
413
51
153
207
248
311
423
330
64
76
111
145
149
192
80
Vet School
Last Name
First Name
Poster
#
University of Florida CVM
University of Georgia CVM
University of Guelph
University of Illinois CVM
Malcolm
Sato
Schrock
Slabaugh
Stromberg
Sundaram
Tenborg
Touitou
Velazquez
Walker
Whisenant
Allgood
Furst
Khan
Kopp
Mills
Mosca
Smith
Stephen
Barnett
Callahan
Cash
Clark
Condrey
Haight
Long
Marinoff
Marshall
Poole
Sirochman
Smith
Turner
Mariampillai
Carter
Dellinger
Engel
Hristova
Ledesma
McLean
Ondera
Pike
Powers
Randall
Reising
Elizabeth
Yu
Kelly
Taylor
Stephanie
Ayswarya
Elizabeth
Florian
Eric
Ashley
Katrijn
Hillary
Nicole
Michael
Rosalind
Kelly
Leandra
Mallory
Alexa
Brian
Robert
Kaycee
William
Jillian
Kimberly
Mackenzie
Jacqueline
Breanna
Tyler
Anna
Katherine
Zachary
Anujah
Samantha
Marion
Danielle
Teodora
Eric
Victoria
Caitlin
Courtney
Jonathan
Chelsea
Abby
220
318
327
340
370
372
383
386
396
407
418
7
114
169
179
250
255
348
361
23
52
58
72
77
128
205
225
226
287
338
346
391
224
57
88
101
148
195
236
274
284
289
297
301
81
Vet School
Last Name
First Name
Poster
#
University of Minnesota CVM
University of Missouri CVM
Storms
Baley
Bancroft
Carlson
CHEN
Daney
Emmitt
Gustafson
Hammes
Ienello
Kuball
Larrabee
Li
Rettkowski
Snyder
Sorg
Tuncay
Wanner
Witschen
Arroyo
Belter
Bishop
Bishop
Burns
Cox
Crombie
Daugherty
Dionne
Ehrlich
Fasig
Girens
Henson
Howard
Huff
Koestel
Landreth
Lind
Mattia
Patterson
Schacher
Schulte
Senft
Siegrist
Slimack
Suzanna
Susan
Channing
Ariel
AOLEI
Jolani
Nicole
Kevin
Joseph
Lauren
Kristin
Shannon
Meng
Rebecca
Kristin
Janna
Mete
Nicole
Patrice
Geraline
Rebecca
Brianne
Kaitlin
Jordan
Amanda
Kathryn
Elizabeth
Madeleine
Allison
Joseph
Renee
Christopher
Jennifer
Jaimie
Zoe
Victoria
Lori
Michael
Delaney
Michael
Rachael
Tyler
Melissa
Katie
365
17
19
55
68
84
100
124
129
156
182
191
197
302
352
356
389
409
425
10
29
36
37
47
79
81
86
92
99
104
121
142
147
150
178
189
201
232
278
320
328
333
337
341
82
Vet School
Last Name
First Name
Poster
#
University of Pennsylvania SVM
University of Saskatchewan WCVM
University of Tennessee CVM
University of Tennessee CVM
University of Wisconsin SVM
Smith
Soltis
Sorenson
Stabler
Stricklin
Swanson
Whittmore
Zorn
Adams
Aguiar
Asklof
Banducci
Binstock
Brockhurst
Clark
Hanes
Hayes
Jimenez
Knebel
Lin
McClung
McGeehan
Meltzer
Nagel
Price
Pruett
Ramos
Ross
Schoenberger
Shearin
Wronski
Lucyshyn
Crilly
Dickson
Belling
Caceres
Dunfield
Falsey
Fedderly
Hausmann
Ludwig
Mack
McMahon
McSweeny
Carolyn
Emily
Juli
Emily
Olivia
Tracy
Jerrianne
Chelsea
Daniel
Ariel
Kendra
Caitlin
Jonah
Jacqueline
Megan
Steven
Allison
Monica
Emily
Megan
George
Megan
Rebecca
Jonathan
Emma
Katherine
Meghan
Taylor
Jenna
Abigail
Sarah
Danica
Nathan
Rachel
Carolyn
Valeria
Elizabeth
Rachel
Galya
Katharine
Allison
Tyler
Rachel
Ian
343
353
355
357
369
373
419
435
3
4
11
20
34
45
71
130
136
159
175
200
234
235
242
262
291
292
296
312
326
334
428
210
80
91
28
50
96
103
105
135
211
216
237
239
83
Vet School
Last Name
First Name
Poster
#
Utrecht University
Utrecht University
VetAgro Sup
Vetagro Sup
Virginia-Maryland Regional CVM
Washington State University CVM
Western College of Veterinary Medicine
Western University of Health Sciences
Myers
Nietlisbach
Nilles
Stilin
Turinski
van Batavia
Vermillion
Weyna
Wierenga
Hendrikx
Smit
Demars
Perier
Caudill
Hecker
Lake
Lawnichak
Mason
Muller
Page
Prado-Sanchez
Russell
Tavella
Zlotnick
Au
Baker
Briggs
Khambholja
Boire
Janse van Rensburg
Timonin
Abdel Massih
Abeywardena
Black
Chan
Kim
Mangosing
Martinez
Mendoza
Nguyen
Ofer
Pador
Richardson
Seelman
Laurel
Nicole
Jacob
Allison
Tiffani
Ashley
Meghan
Alisia
Lauren
Lysanne
Michelle
Fanny
Nadege
Mitchell
Kari
Bathilda
Tyler
Caitlin
Stefanie
Lauren
Evymarie
Ellen
Vincent
Marta
Gina
Anna
Cassidy
Chantelle
Francois
Claire
Mary
Cherein
Shanes
Kelley
Dennis
Sohyun
Sara
Aileen
Gerardo
Steven
Oren
Sean
Ella
Amanda
261
266
267
364
390
393
398
417
421
140
342
89
280
62
139
186
194
230
258
276
290
315
381
434
12
16
44
168
41
157
385
1
2
38
63
171
221
228
244
264
269
275
305
332
84
Vet School
Last Name
First Name
Poster
#
Tang
Voss
Karena
Brittany
376
403
85
Young Investigator Award Finalist Abstracts
Development of a genetic risk profile for Epstein-Barr virus–associated post-transplant lymphoproliferative disease
Elshafa H. Ahmed, Robert A. Baiocchi, and Michael A. Caligiuri
Comparative and Veterinary Medicine Program, College of Veterinary Medicine (Ahmed), and Division of Hematology,
Department of Internal Medicine (Baiocchi, Caligiuri), The Ohio State University, Columbus, OH
Redacted
86
Unraveling the genetic basis of the immune response to pertussis vaccination
Yung-Yi C. Mosley, Josiah Radder, Annerose Berndt, and Harm HogenEsch
Department of Comparative Pathobiology, College of Veterinary Medicine, Purdue University,
West Lafayette, IN (Mosley, HogenEsch); and Department of Medicine, School of Medicine, University of Pittsburgh
Pittsburgh, PA (Radder, Berndt)
Whooping cough (pertussis) is a highly contagious bacterial disease caused by Bordetella pertussis. Pertussis is prevented
by vaccination with diphtheria-tetanus-acellular pertussis (DTaP) vaccines for children younger than age 7. In spite of high
vaccine coverage, the incidence has increased in children, adolescents, and adults in recent years, with nearly 110,000 cases
reported in the US during 2012-2014. Rapid waning of immunity induced by vaccination is thought to play a major role in
recent epidemics. Twin studies have shown that vaccine-induced immune responses are heritable and the genetic variation
between individuals can contribute to the heterogeneity of the immune responses to childhood vaccines. Hence, apart from
developing a new pertussis vaccine, we performed a genome-wide association study (GWAS) in mice to reveal the genetic
basis that could control the immune response to the current pertussis vaccine. Female mice of 27 inbred strains were vaccinated with DTaP at 6, 8, and 12 weeks of age. Total IgG antibody titer against the three pertussis antigens in the DTaP
vaccine including pertussis toxin (PT), filamentous hemagglutinin (FHA), and pertactin (Prn) were measured by ELISA at
weeks 14 and 24 of age. Antibody titer at week 14 represents the magnitude of the antibody response, and antibody longevity was calculated by IgG antibody titer at week 24 divided by that of week 14 of age. There were marked differences
in the immune responses among mouse strains, indicating that genetic variability contributes significantly to the magnitude
and duration of anti-pertussis IgG responses. The magnitude and longevity of antibodies induced by DTaP vaccine were not
significantly correlated. Comparison of IgG responses to PT, FHA, and Prn revealed stronger and more significant correlations for longevity than for antibody magnitude, suggesting that different mechanisms drive the longevity and magnitude
of the IgG response. Haplotype association mapping identified common genetic markers on chromosomes associated with
antibody longevity. Utilizing PolyPhen analysis to predict functional effects of single nucleotide polymorphism (SNP), six
genes were identified that potentially control antibody longevity following DTaP immunization. On the other hand, there
were no common genes that affected the magnitude of the antibody response to the three pertussis antigens. Further experiments are aimed at confirming the expression of the identified genes involved in antibody longevity by RT-qPCR as well as
determining the role of these candidate genes in shaping the antibody longevity. We will also investigate whether noncoding
RNAs are involved in controlling the immune responses to pertussis vaccination by examining the transcriptomes of the
draining lymph node in short- and long-longevity mouse strains. We expect that this research will uncover mechanisms that
shape vaccine longevity. This information could be further translated into personalized DTaP vaccination schedules as well
as improved vaccine formulations that induce a more effective immune response.
87
Microbiome associations in pigs with the best and worst clinical outcomes following coinfection with porcine
reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2)
Megan C. Niederwerder,, Crystal J. Jaing, James B. Thissen, Ada Giselle Cino-Ozuna, Kevin S. McLoughlin,
and Raymond R. R. Rowland
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University, Manhattan,
KS (Niederwerder, Cino-Ozuna, Rowland); and Physical & Life Sciences Directorate (Jaing, Thissen) and Computations
Directorate (McLoughlin), Lawrence Livermore National Laboratory, Livermore, CA
On a worldwide basis, coinfections involving porcine reproductive and respiratory syndrome virus (PRRSV) and porcine
circovirus type 2 (PCV2) are common and contribute to a range of polymicrobial disease syndromes in swine. Both viruses
compromise host defenses, resulting in increased susceptibility to infections by primary and secondary pathogens that can
affect growth performance as well as increase morbidity and mortality. An experimental population of 95 nursery pigs was
coinfected with PRRSV and PCV2. At 70 days postinfection (dpi), 20 representative pigs were selected as having the best
or worst clinical outcome on the basis of average daily gain (ADG) as well as the presence and severity of clinical disease.
Worst-clinical-outcome pigs had prolonged and greater levels of both PRRSV and PCV2 viremia as measured by qPCR.
Serum, lung, and fecal samples collected at 70 dpi were analyzed using a comprehensive DNA microarray technology, the
Lawrence Livermore Microbial Detection Array, which can detect over 8,000 microbes. Bacterial species, such as Bacillus
cereus, were detected at a significantly higher rate in the serum of worst-performing pigs. At the level of the fecal microbiome, the overall microbial diversity was significantly lower in the worst-clinical-outcome group. Members of the Proteobacteria phylum, specifically Escherichia coli, were detected at a significantly higher rate in the feces of best-performing
pigs. The results reinforce the importance of pathogen load in determining clinical outcome and suggest an important role of
microbial diversity as a contributing factor in disease. With growing pressure to eliminate antibiotic use and innovate new
management tools for infectious diseases in food animals, this work provides foundational knowledge about the microbiome
characteristics that may contribute to performance enhancement in the presence of common viral pathogens.
88
Characterization of immunomodulatory properties of feline mesenchymal stem cells
M. Parys, J. M. Kruger, and V. Yuzbasiyan-Gurkan
Department of Small Animal Clinical Sciences and Comparative Medicine and Integrative
Biology Program, College of Veterinary Medicine, Michigan State University, East Lansing, MI
Introduction: Inflammatory diseases are a major burden in both human and veterinary
medicine. Mesenchymal stem cells (MSCs), through their soluble and membrane-bound factors, are capable of affecting the
majority of inflammatory cells in the body, inducing their anti-inflammatory properties or blocking expansion/proliferation.
These factors include IDO1, iNOS, COX2, TGF-β1, HGF, IL-6, PD-L1, FASL, HMOX1, and IL-10. Cats spontaneously
develop a wide variety of inflammatory disorders such as asthma, inflammatory bowel disease, chronic hepatitis, chronic
kidney disease, and chronic idiopathic cystitis. These diseases may benefit from MSC-based therapies and may serve as
translational models for human disorders. Interestingly, interspecies differences between expressions of these genes exist.
For example, human MSC immunomodulation is considered to be IDO1 dependent, while iNOS is the key gene involved
in immunomodulation in mice. The aim of this project was to evaluate the immunomodulatory properties of feline MSC.
Materials and Methods: MSCs were isolated from adipose tissue specimens collected from purpose-breed cats. After collagenase I dissociation, cells were plated and expanded in KNAC medium supplemented with 5% FBS. Propagated cells
were characterized for expression of cell linage markers CD90, CD44, CD105, and MHCII, as well as trilineage differentiation. Cells from passages 3-6 were exposed to increasing amounts of INFγ (0.25 ng/mL to 50 ng/mL) and TNFα (0.1 ng/
mL to 10 ng/mL) for 24 hours. RNA was then extracted, quantified, and reversed transcribed using reverse transcriptase.
Relative expression of targeted genes involved in immunomodulation was determined by quantitative RT-PCR. Protein levels of IL-6, IL-10, TGF-β1, HGF, and PGE2 were quantified by ELISA. IDO1 activity was measured by Ehrlich’s assay after
3-day stimulation with INFγ. MSC-mediated inhibition of proliferation of ConA-stimulated peripheral blood mononuclear
cells was assessed by co-culture experiments. For these experiments, cells were plated at a 1:5 ratio of irradiated MSC to
effector cells in a 96-well plate and incubated for 72 hours. EdU, a thymidine analogue, was added for the last 24 hours of
the incubation period, and cell proliferation was measured using fluorescence plate reader.
Results: All experimental MSC lines strongly expressed CD90, CD44, and CD105 markers and differentiated into osteocytes, chondrocytes, and adipocytes. Unstimulated MSC expressed all immunomodulatory genes, except for IL-10 and
FASL. Levels of iNOS were low or undetectable at the RNA level. Four genes were significantly upregulated in a dose-dependent manner after INFγ stimulation: IDO1, IL-6, HGF, and PD-L1. After TNFα stimulation, only IL-6 expression was
increased. At the protein level, significant increases were detected in IL6 after stimulation with a single dose of TNFα and
INFγ as well as after TNFα and INFγ co-stimulation. Levels of HGF were significantly decreased after stimulation with
INFγ as well as both TNFα and INFγ. Three-day stimulation with INFγ resulted in significant increase in IDO produced
kynurenine in cell culture medium. In co-culture experiments, incubation of MSC with stimulated PBMCs significantly
blocked proliferation.
Conclusions: Feline MSCs are similar to human MSCs in their reaction to inflammatory stimuli and are capable of successfully blocking proliferation of stimulated PBMCs in co-culture experiments. Feline MSCs more closely parallel human
MSCs in their immunomodulatory properties than mouse MSCs and thus may be more appropriate models to study MSCbased therapies.
89
Biomaterial composition influences in vitro macrophage behavior
N. Springer and C. Fischbach
Biological and Biomedical Sciences Program, College of Veterinary Medicine (Springer, Fischbach), and Nancy E. and
Peter C. Meinig School of Biomedical Engineering (Springer, Fischbach), Cornell University, Ithaca, NY
Introduction: Mechanical perturbations to cells and tissue, mediated by extracellular matrix (ECM) remodeling, are an
established contributor to disease processes like tumorigenesis. The influence of biophysical forces on cancer cells is well
documented, but the role of mechanical regulation on other key cells in tumorigenesis, such as immune cells, has been largely ignored. Classically, immune cell function is examined independently of the microenvironment, either via flow cytometry
or by in vitro experiments performed on nonphysiologic tissue culture plastic. As immune cells, such as macrophages, adapt
their behavior to a variety of stresses and tissue elasticities during invasion and migration, mechanosignaling, the transduction of mechanical stimuli into biochemical signals that guide cellular behaviors, is likely important in immune cell
activation in vivo.
Objective: Elucidate the role of ECM biophysical and biochemical input on macrophage behavior utilizing synthetic and
natural biomaterial models.
Materials and Methods: Bone marrow–derived macrophages (BMDM) isolated from C57Bl/6 mice were cultured on fibronectin-functionalized polyacrylamide (PA) hydrogels mimicking the bulk stiffness of compliant (140 Pa), intermediate
(1 kPa), and fibrotic (10 kPa) tissues; decellularized native ECM assembled by adipose stromal cells isolated from lean
(less stiff) or obese (more stiff) mice; or fibronectin functionalized glass (gPa) to reflect standard tissue culture techniques.
BMDM proliferation (BrdU), morphology (aspect ratio), and polarization state (LPS or IL-4/13 stimulation and inducible
nitric oxide synthase [iNOS] or arginase-1 [Arg-1] production) were evaluated via immunofluorescence and image analysis.
Results: BMDM proliferation positively correlated to stiffness on PA gel and glass platforms with significant differences (p<0.05) in BrdU incorporation between all stiffnesses. Unexpectedly, BMDM proliferation was significantly greater
(p<0.05) on lean vs. obese ECM. Disruption of mechanosignaling by Y27632, a Rho-associated kinase inhibitor, significantly decreased proliferation; however, BMDM on native ECM were less sensitive to inhibition. Average aspect ratio did not
differ across PA gels, although total cell area was significantly increased on 10-kPa gels, indicating greater cell spreading
and tension on stiffer substrates. BMDM on glass had the highest aspect ratio and cellular area of all conditions. Average aspect ratio was similar in BMDM on lean and obese ECM, but BMDM on lean ECM had a much wider range of aspect ratios
indicating differences in BMDM adhesion. After stimulation to classical (LPS) or alternative (IL-4/13) polarized states, both
iNOS and Arg-1 immunoreactivity significantly decreased (p<0.05) with increasing stiffness including a marked decrease
in the glass condition. Unexpectedly, iNOS expression was significantly increased on obese vs. lean ECM; results for Arg-1
expression on native ECM are pending.
Conclusions: Our results indicate that compositional, in addition to mechanical, properties of biomaterials for in vitro model systems are integral to macrophage
behavior. Selection of biomaterials to recapitulate physiological conditions must be carefully considered in experimental
design and interpretation. These findings are broadly relevant to fields where investigating macrophage behavior is critical,
such as tumor microenvironment, immune-mediated disorders, regenerative medicine, and biomedical device development.
Experiments are underway to deconstruct the biophysical and biochemical input from native ECM on BMDM by coupling
decellularized ECM to the polyacrylamide hydrogel system.
Acknowledgements: Supported by NIH Awards #T32OD0011000 and #1R01CA185293, and the Cornell Center on the
Microenvironment and Metastasis, Award #U54CA143876
90
Complete Abstracts
Cherein Abdel Massih, Daniel Avenick, Stephanie Istvan, Keith Richter, Nicole Edwards, Nigel Mackman,
and Linda Kidd
Western University of Health Sciences, College of Veterinary Medicine, Pomona, CA (Abdel Massih, Kidd);
Veterinary Specialty Hospital, San Diego, CA (Avenick, Istvan, Richter, Edwards); University of North Carolina
at Chapel Hill (Mackman)
Microvesicles (MVs) are membrane fragments derived from cells including RBCs, platelets, and leukocytes. Tissue factor
(TF+) and phosphatidylserine (PS+) expressing MVs are procoagulant. They are released during hemolytic and proinflammatory states in-vivo. MVs also accumulate in packed RBCs (PRBC) during storage. We have shown that procoagulant
activity (PCA) associated with TF+ and PS+ MVs is increased in dogs with IMHA. Transfusion of aged PRBCs is associated with mortality in dogs with hemolysis. Thus MV PCA may contribute to thrombosis and inflammation in these patients.
We hypothesize that PS+ and TF+ MV PCA increases in canine(c) PRBC units during storage and leukocyte reduction
(LR) reduces its accumulation. Our objectives were to establish whether a centrifugation protocol that minimizes platelet
contamination and maximizes MV yield from plasma results in acceptably low platelet numbers in supernatant from cPRBCs, determine if a plasma based PS capture thrombin generation assay (Zymuphen MP-Activity Aniara) detects PS+MV
PCA in cPRBC supernatant, and to measure PS+MV and TF+MV PCA in LR and non-LR cPRBCs. Based on a previously
described semiquantitative platelet contamination score (PCS), residual platelet levels in cPRBC supernatant were similar
to citrated plasma (cPRBC n=9; mean PCS 0.2557 +/- 0.27024; citrated human plasma n=10; published mean PCS 0.25 +/0.4; p=0.9173). The PS+MV PCA assay detected PCA in cPRBC supernatant. Dilutions ranged from 1:20 for samples taken
the day of donation to 1:5,000 for samples from an expired hemolyzed unit. 5 LR and 5 NLR cPRBC units are currently
being sampled over 42 days to determine whether PS+ and TF+MV PCA increase during storage and whether it is affected
by LR.
Research Grant: WesternU Office of Research and Biotechnology, College of Veterinary Medicine Clinical Partnership
Matching Grant, and Veterinary Specialty Hospital.
Student Support: WesternU CVM Veterinary Scholars Program and WesternU Summer Research Fellowship Program.
Shanes C. Abeywardena and David C. Kersey
Western University of Health Sciences CVM, Pomona, CA
Maintaining health and reproduction is critical for sustaining threatened species in captivity. Because the endocrine system
helps maintain these functions, understanding basic physiology of metabolic and reproductive activity through hormone assessment allows management to meet conservation goals for the species. The objective of this study was to understand the
biologic activity of giant panda, red panda, and killer whale species via endocrine techniques. For the giant panda (Ailuropoda melanoleuca) an artificial insemination was performed in the spring of 2016. To track the hormone changes associated
with the variable and unique luteal phase we assessed urinary estrogens and progestagens. In the red panda (Ailures fulgens)
a lack of basic knowledge of male reproductive activity has hampered captive breeding efforts. This study validates an approach to assess androgen values in the feces of male red pandas and track circannual changes in testicular activity. In the
killer whale (Orcinus orca), little knowledge exists about the metabolic activity of the species. For this study we validated
thyroid hormone assays in the assessment of banked serum, with the goal of designing an approach to create a demographic
database of thyroid hormone levels for the species. From these combined studies we applied techniques for sample handling
and hormone extraction, immunoassay theory and methodology, validation of steroid and thyroid hormones from various
sample types, and assessment of hormone data in a biological context. This study demonstrates the broad value endocrine
evaluation provides for species conservation and the various techniques necessary to accomplish those goals.
Research Grant: None
Student Support: WesternU Summer Research Fellowship; CVM Veterinary Scholars Program
91
Determining the effects of SMAC mimetic drugs on vascular endothelial cells
Daniel J. Adams, Kelly A. McCorkell, and Michael J. May
Department of Biomedical Sciences, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA
Members of the Nuclear Factor-kB (NF-kB) family of transcription factors play crucial roles in regulating inflammation
and immune function. NF-kB can be activated by two different mechanisms: the classical and non-canonical pathways.
The non-canonical pathway is vital for immune function and development of secondary lymphoid organs. Aberrant non-canonical NF-kB contributes to autoimmune diseases such as ulcerative colitis and rheumatoid arthritis. In contrast to the
classical NF-kB pathway, the spectrum of genes regulated by non-canonical NF-kB is poorly defined. Vascular endothelial
cells (VECs) are a major cell type involved in immune and inflammatory responses. We previously showed that ligation of
the Lymphotoxin-b receptor (LTbR) by LIGHT induces gene expression via the non-canonical pathway in VECs. Our lab
has also found that a novel class of anti-cancer drugs named second mitochondria-derived activator of caspase (SMAC)
mimetics (SMs), also simulate non-canonical NF-kB in VECs. A crucial question that remains regarding the efficacy of
SMs centers on their ability to activate non-canonical NF-kB in normal cells. The goal of this study is to determine the effects of SM drugs on non-canonical NF-kB-driven gene expression in VECs. We stimulated human aortic endothelial cells
(HAEC) with TNFa, LIGHT, and the SM drug GT13072. We found by microarray that the SMs induces expression of a
range of pro-inflammatory and pro-angiogenic genes. We are currently establishing which of these genes are activated via
the non-canonical NF-kB pathway and we will pursue functional studies to determine if these genes contribute to inflammatory and angiogenic responses of VECs.
Research Grant: NIH 2R56HL096642-05
Student Support: NIH-Merial Veterinary Scholars Program
Characterizing immune regulation of canine heartworm Dirofilaria immitis infection in Aedes aegypti mosquitoes
Ariel M. Aguiar, James B. Lok, Thomas J. Nolan, Letitia K. Thompson, and Michael Povelones
Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA
Heartworm infection in canids and felids is caused by the parasitic nematode Dirofilaria immitis. D. immitis is transmitted
between hosts by mosquito vectors, which are required intermediate hosts for the parasite, supporting development from
the microfilaria to the infective third-stage larva. Heartworm is an important disease in North America due to its high prevalence. Due to evidence of resistance to heartworm preventatives, it is necessary to investigate other transmission control
methods. One such approach is targeting the mosquito vector itself. Aedes aegypti is a well-studied laboratory mosquito
model for D. immitis infection. Strains that are susceptible or refractory to infection are available. Preliminary data from our
lab suggests that genetically hyperactivating the REL1/NF-kB immune signaling pathway significantly reduces D. immitis
infection in susceptible mosquitoes. Here we test the hypothesis that REL1, or another related factor, REL2, contributes to
the natural resistance of a refractory Ae. aegypti strain. To test this, REL1 and REL2 were genetically silenced in refractory
mosquitoes by RNA-interference. Mosquitoes were then fed on blood containing infective D. immitis microfilaria. One
week post infection, mosquitoes were dissected, fixed, and fluorescently stained to assess the number, location, and development of the worms. Our data shows there is no difference between the mock-treated control group and those where REL1
or REL2 was silenced. We are currently measuring the gene silencing efficiency by quantitative RT-PCR. Based on our
preliminary data, we conclude that resistance to D. immitis infection in the refractory Ae. aegypti is likely not due to REL1
or REL2 signaling.
Research Grant: School of Veterinary Medicine Intramural Funding
Student Support: Grant from Merial Ltd. and NIH grant T35 OD-010919
92
Socioeconomic status of population and negligence of dogs with visceral leishmaniasis in Araçatuba-SP, Brazil
Derecka Jamie Alexander, Livia Castanhas Bregano, Hugo Ribeiro, Tamiris Firmiano de Jesus, Moara Martins,
Ingeborg Langohr, Gisele Fabrino Machado
School of Veterinary Medicine, Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State
University, Baton Rouge, LA, (Alexander, Martins, Langohr), Univ. Estadual Paulista- UNESP, Araçatuba, SP, Brazil
(Bregano, Ribeiro, Jesus, Machado)
Visceral leishmaniasis (VL) can lead to weight loss and emaciation in dogs, and the presence of other infectious diseases
associated with malnutrition can contribute to worsening of clinical signs. The law in Brazil determines that in the case of
a positive diagnosis of VL, the dog must be euthanized, with the intention to protect the human population. Treatment is
not recommended because it does not lead to a clinical cure. Moreover, there is no evidence of complete elimination of the
parasite. Nonetheless, despite violating the law, owners decline euthanasia procedures performed by the Zoonosis Control
Center (CCZ) of Araçatuba -- SP, and continue to house sick dogs. Seropositive dogs from the CCZ were grouped as asymptomatic/oligosymptomatic and multisymptomatic. We performed measurement of bone marrow (BM) fat at necropsy in
order to verify the existence of cases suggestive of animal negligence. We then correlated these data with a map of the socioeconomic status of the population. Our hypothesis was that dogs with low BM fat content would be found at boundaries of
the city, where the diagnosis of VL is more prevalent and the socioeconomic conditions are poor. Among 19 dogs evaluated
at necropsy, all presented BM fat content lower than 82%. Five of them were multisymptomatic dogs with less than 20% of
BM fat, collected in areas with better socioeconomic conditions. These results, even though preliminary, can serve as a basis
for the development of educational policies or punitive action aimed to reducing the mistreatment of animals.
Research Grant: São Paulo Research Foundation (FAPESP) 2016/02384-9
Student Support: National Institutes of Health (NIH)
Flow cytometric evaluation of canine lymphocytes after oral mycophenolate mofetil
Kayla Alexander, Charlee Mulligan, Barbara Kaplan, Andrew Mackin
Department of Clinical Sciences (Alexander, Mulligan, Mackin) and Department of Basic Sciences (Kaplan),
College of Veterinary Medicine, Mississippi State University, Mississippi State, MS
Mycophenolate mofetil (MMF) is an immunosuppressive agent that has been used in human medicine for organ transplant
recipients. The low cost of MMF has led to an increase in use in dogs for the treatment of autoimmune disease. MMF
modulates immune responses by inhibiting inosine-5’-monophosphate dehydrogenase (IMPDH), an enzyme needed for the
synthesis of purines. Two isotypes of IMPDH exist: IMPDH1 and IMPDH2. IMPDH2 is used exclusively by lymphocytes
during proliferation, whereas other cells are able to utilize a salvage pathway for the synthesis of purine nucleotides. Inhibition of IMPDH2 selectively inhibits lymphocyte mitosis, leaving cells unable to replicate. Researchers hypothesize that the
immunomodulatory effects of MMF are mostly due to an inability of lymphocytes to proliferate. Other studies suggest that
MMF also induces lymphocyte apoptosis. Ongoing studies in our laboratory have documented that MMF, after two weeks
of administration, inhibits lymphocyte proliferation. This current study was designed to use flow cytometry to determine
whether MMF in dogs, at maximally tolerated doses for two weeks, also causes lymphocyte death and apoptosis. In addition, flow cytometry was used to determine the effects of MMF on canine CD4+ and CD8+ lymphocyte populations. Our
results indicated that, compared to pretreatment values, oral MMF for two weeks had minimal effect on lymphocyte apoptosis (early or late), cell death, or populations of CD4+ and CD8+ cells. Our findings support the theory that MMF inhibits
lymphocyte proliferation primarily by causing cell cycle arrest, rather than cell apoptosis or death.
Research Grant: Mississippi State University College of Veterinary Medicine
Student Support: Morris Animal Foundation
93
Surveillance of Florida snails for Parelaphostrongylus andersoni and Parelaphostrongylus tenuis
Hillary E. Allgood, James F. Wellehan, Jr., Linda Archer, Heather S. Walden
Department of Small Animal Clinical Sciences (Allgood, Wellehan, Archer) and Department of Infectious Diseases and
Pathology (Walden), College of Veterinary Medicine, University of Florida, Gainesville, FL.
Metastrongyloid nematodes of the genera Parelaphostrongylus are common parasites of ruminants, specifically cervids,
which have a widespread distribution throughout North America. Parelaphostrongylus andersoni and P. tenuis both use the
white-tailed deer (Odocoileus virginianus) as their definitive host; terrestrial gastropods are intermediate hosts. Prevalence
of Parelaphostronglyus species in Florida has not been well documented. P. tenuis is prevalent in eastern North America,
but has only been found to inhabit Florida in 1972, when it was found in Collier County among a population of relocated
white-tailed deer from Wisconsin (Prestwood and Smith 1969). It was hypothesized that P. andersoni occurs in greater
numbers in Florida than what has been reported. The specific aim of this study was to survey 1,346 gastropods, comprising
of 25 species, from 15 counties in Florida for the presence of P. andersoni and P. tenuis. A quantitative polymerase chain
reaction (qPCR) assay targeting the cytochrome oxidase I (COI) gene, which was specific to each Parelaphostrongylus spp.,
was developed. qPCR data was used to estimate the prevalence and distribution of Parelaphostrongylus spp. in Florida, as
well as to determine specific snail species that can act as intermediate hosts. qPCR assays are currently being investigated
at the time of submission of this abstract.
Research Grant: Cervidae Health Research Initiative (CHeRI), Department of Wildlife Ecology and Conservation, Institute of Food and Agricultural Sciences, University of Florida, Gainesville, FL.
Student Support: Merial
The impact of a cancer-specific diet on inflammation and side effects associated with chemotherapy in dogs
Jessica S. Anderson and Korinn E. Saker
Department of Molecular Biomedical Sciences, College of Veterinary Medicine, NC State University, Raleigh, NC
Up to 30% of canine chemotherapy patients can experience significant side effects which negatively impact their gastrointestinal (GI) function, appetite, body weight and energy level, ultimately resulting in a diminished quality of life (QOL). A
plethora of research on the nutrient and functional food aspects of nutritional support for the human cancer patient exists;
however, evidence-based nutritional guidelines for the canine cancer patient are limited. The objectives of this study are to
evaluate the impact of a canine cancer-specific (CCS) diet on systemic and GI inflammation in dogs diagnosed with lymphoma and receiving chemotherapy; and to determine correlations between their inflammatory state and severity of side effects
experienced during treatment. Thirty client-owned dogs diagnosed with lymphoma and receiving chemotherapy treatment
through the oncology service at NC State Veterinary Hospital will be identified and enrolled for the study. Each dog will be
fed both the CCS diet and their normal home diet on an alternating schedule over a 3-month period. Owners will complete
a weekly feedback survey on their pet’s clinical symptoms and QOL. Additionally, blood and fecal samples will be obtained
for inflammatory marker analysis. Commercial canine C-reactive protein (CRP) and fecal calprotectin ELISA assays will
be utilized to measure systemic and GI inflammation, respectively. Assay reactivity will be evaluated prior to study sample
analysis to ensure each assay selected is appropriate for the study. The preliminary data for the CRP ELISA reactivity in
healthy, chronic and acutely inflamed dogs yielded 49928.12, 60007.03, 146952.70 ng/ml, respectively. The study is in
progress, so final conclusions are pending.
Research Grant: Canine Biologics LLC
Student Support: Merial Veterinary Scholars Program
94
Investigating the effects of EZH2 inhibition in canine B-cell lymphoma
Andrea E. Arica, Fiona Hennig, Corri Levine, Angela McCleary-Wheeler
Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY
Canine B-cell lymphoma is one of the most common malignancies in the dog. While treatable, it is often incurable. Canine
B-cell lymphoma has many similarities to human diffuse large B-cell lymphoma (DLBCL), but one area less studied in the
dog is the histone landscape. EZH2 is the catalytic subunit of the polycomb repressive complex 2, which is involved in
the transcriptional silencing of chromatin. EZH2 represses target gene expression by trimethylating histone H3 on lysine
27 (H3K27me3). Overexpression of or activating mutations in EZH2 contributes to the tumorigenesis of human DLBCL.
Increased EZH2 activity causes a global increase in H3K27me3. Several EZH2 target genes are involved in cell cycle regulation; therefore, their repression leads to unchecked cell proliferation and tumor development. Studies in human DLBCL
suggest that inhibition of EZH2 results in inhibition of cell proliferation. To assess the effects of an EZH2-specific inhibitor,
JQ EZ-005, on EZH2 activity in canine B-cell lymphoma, the canine B-cell malignancy cell lines CLBL-1 and 17-71 were
utilized. Western blots were performed to confirm a global decrease in the H3K27me3 mark in the treated cell lines. Quantitative reverse-transcription PCR was carried out to evaluate the effect of EZH2 inhibition on the expression of several
selected target genes. Flow cytometry was used to assess the cell cycle status of the treated cell lines. Evaluation of EZH2
activity in canine B-cell lymphoma will provide added insight into the epigenetic factors involved with this cancer and potentially lead to the development of novel therapeutic strategies that can be used to treat both canine and human lymphoma.
Student Support: NIH T35 Training Grant
Comparison of MALDI-TOF and PFGE for strain-typing Staphylococcus aureus isolated from cow’s milk
Geraline T. Arroyo, Pamela R. F. Adkins, John R. Middleton
Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri,
Columbia, Missouri
Staphylococcus aureus is one of the most common contagious mastitis pathogens of dairy cattle. Subspecies strain-typing is
used to differentiate contagious strains from sporadic ones within a herd to aid in management. The current “gold standard”
for strain-typing is pulsed-field gel electrophoresis (PFGE), but it is relatively expensive and time consuming. Matrix assisted laser desorption ionization time of flight (MALDI-TOF) mass spectrometry identifies bacterial isolates based on each
isolate’s ribosomal proteome and has been shown to be more cost effective and less time consuming for speciating bacterial
isolates. In addition, recent studies suggest that variability between spectrograms within a given genera and species of bacteria may be used to strain-type isolates. The purpose of this study was to determine whether the MALDI-TOF could differentiate strains of S. aureus isolated from cow’s milk with the same discriminatory power as PFGE. A total of 65 S. aureus
isolates from 8 dairy herds that had already been characterized by PFGE were studied. Isolates were cultured on blood agar
and prepared for MALDI-TOF analysis using the manufacturer’s described extraction method. A total of 24 spectra were
obtained for each isolate. From these spectra, a consensus spectra was created using MALDI Biotyper OC. PFGE data and
MALDI-TOF data were compared using Bionumerics 7.5. To date, consensus spectra have been obtained from 37 isolates.
Preliminary data shows clustering within farm. While some isolates clustered according to the previously determined PFGE
type, some did not. Further work is needed to determine if MALDI-TOF strain-typing is as discriminatory as PFGE and able
to define contagious strains of S. aureus.
Research Grant: USDA National Institute of Food and Agriculture, Animal Health project # 1008624
Student Support: USDA National Institute of Food and Agriculture, Animal Health project # 1008624
95
Kendra P. Asklof, Lance W. Peterson, Naomi H. Philip, Igor E. Brodsky
University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA, USA
Yersinia are pathogenic bacteria that infect animals and humans, causing disease such as severe gastroenteritis and plague.
Yersinia inhibits NF-kB and MAPK signaling pathways in infected cells, reducing pro-inflammatory cytokine release and
inducing cell death pathways. These cell death pathways are thought to be important for host protection during Yersinia
infection, but the precise mechanism of how cell death contributes to immune defense is unclear. The primary cell death
pathway involves signaling between RIPK1 and caspase-8, inducing apoptosis of infected host cells. In the absence of
caspase-8, another pathway involving a RIPK1 and RIPK3 complex causes necroptosis, suggesting a role as a back-up
pathway to ensure cell death during Yersinia infection. While the presence of caspase-8 inhibits RIPK3 activation, RIPK3
knockout mice succumb to Yersinia infection approximately 6-8 days following oral infection. This suggests that RIPK3
is not only a back-up cell death pathway, but rather is essential for host survival during Yersinia infection. In an effort to
illuminate RIPK3’s role during Yersinia infection, we analyzed CFU and cytokine expression through flow cytometry in
various organs harvested from RIPK3 -/- and wildtype mice at day 5 of infection. Interestingly, we observe no difference
in CFU levels in spleens, mesenteric lymph nodes, livers, or peyer’s patches and no difference in cytokine expression of
monocytes, neutrophils, or dendritic cells in spleens or mesenteric lymph nodes. This indicates that RIPK3 does not play a
role in controlling bacterial burdens or innate cytokine levels at this early time point during Yersinia infection, making its
role in controlling resistance to Yersinia as-yet undefined.
Research Grant: R01-AI103062 and R21-AI109267
Student Support: NIH T35-OD010919 and a grant from Merial
Combinatory radiotherapy and cellular immunotherapy enhances tumor cell killing
Gina G. Au, Laura L. Bronsart, Edward E. Graves
Department of Radiation Oncology, Stanford University, Stanford, CA
Natural killer (NK) cells have been shown to play an important role in the tumor immune response, and leukemic NK cell
lines are currently being developed as cellular immunotherapeutics. In this study, we investigated the outcome of combining
NK cell therapy with ionizing radiation to specifically target cancer cells. The natural killer group 2D (NKG2D) receptor
is expressed by all NK cells, and its ligands are induced by cellular stress. We hypothesized that ionizing radiation would
increase the cell-surface expression of NKG2D ligands in the breast cancer (MDA-MB-231) and lung adenocarcinoma
(A549) cell lines, enhancing their ability to activate NK cell-mediated killing. Flow cytometry revealed an increase in the
cell-surface expression of NKG2D ligands and an additional NK activation ligand, CD48, on the breast cancer cells following radiation. However, the effect in the lung adenocarcinoma cells was not as pronounced. We also tested the cytotoxic
effects of the NK leukemic cell line, NK92, and found a reduction in breast cancer cell survival upon addition of NK92 cells.
This effect was enhanced when NK92 cells were applied to irradiated cancer cells. These findings suggest that radiation
can induce NK-activating ligand expression on breast cancer cells, possibly making them more susceptible to the cytotoxic
effects of NK cells. By identifying the impact of irradiation on NK-activating ligands, we hope to develop a more targeted,
dual-modality approach to combatting cancer.
Research Grant: NIH R01: Radiation-Induced Tumor Cell Migration
Student Support: NIH Office of the Director, Division of Comparative Medicine
96
Rebecca Aust, Caitlin Anglin, Claire Birkenheuer, Shannon Dehghanpir, Kui Yang, and Joel Baines
Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA
Upon infection with herpes simplex virus 1, host cell RNA polymerase II (RNA Pol II) is altered to begin transcription of
viral mRNAs. In uninfected cells, RNA Pol II utilizes cyclin dependent kinase 9 (CDK9) for transcription progression. It
has been suggested that CDK9 is important for RNA Pol II to function after HSV1 infection by use of the non-specific CDK
inhibitor Flavopiridol. We hypothesized that CDK9 is an important component for RNA Pol II to properly transcribe HSV1
mRNA. This hypothesis predicts that CDK9 inhibitors such as Dinaciclib, Flavopiridol, and SNS-32 will reduce HSV1
replication and transcription. To test this hypothesis, CV1 cells were mock infected or infected with wild type HSV1 in
the presence of growth media containing subtoxic doses of the drugs or DMSO only. Viral replication was then measured
by plaque assay and gene expression was measured by Western blot with HSV-specific antibodies. The cells that were infected and treated with the CDK9 inhibitors showed a 15-100 fold reduction in the number of plaques when compared to
the DMSO control, indicating that the CDK9 inhibitors reduced viral replication. Similarly, the Western blots revealed a
reduction of the late stage viral infection protein UL-51 in the infected, drug treated samples indicating that gene expression
was reduced. These findings indicate that the CDK9 protein is playing an important role in the transcription of HSV1 and
suggest a new set of drugs that may be effective against herpesviruses.
Research Grant: Unknown
Student Support: Louisiana State University School of Veterinary Medicine Summer Scholars Program
Study of human and mouse lung cells responses to carbon nanotubes to assess their pulmonary disease potential
Barbara J. Avalos-Cavero, Alexia J. Taylor, James C. Bonner
College of Veterinary Medicine, North Carolina State University, Raleigh, NC. (Avalos-Cavero) Department of
Biological Sciences, North Carolina State University, Raleigh, NC. (Taylor, Bonner)
The innovative use of carbon nanotubes (CNTs) has been widely increasing in the nanotechnology industry. Their use in
drug delivery, cancer theragnosis, and the fabrication of batteries and structural materials has steered researchers to further
study their applications in science and technology. A way to enhance their unique properties is by means of functionalization. This process has led to the fabrication of numerous types of CNTs whose biological effects and potential toxicity are
still unknown. Previous studies done in rodents have demonstrated that CNTs have the potential to cause immunotoxic and
fibroproliferative responses in the human body. We aim to study the toxic effects that a group of purified and functionalized
multi-walled carbon nanotubes (MWCNTs) have on a human bronchial epithelium cell line, BEAS-2B, and a mouse alveolar type II epithelial cell line, E10, by exposing the cell lines to different dosages of MW-CNTs within a 24hr time frame.
Preliminary results have shown that BEAS-2B cells exposed to a dose of 100 mg/ml of chemically purified MWCNTs (NC3100) or carboxyl functionalized MWCNTs (NC-3151), exhibit less than a 0.5% cytotoxicity when analyzed using a Lactate
Dehydrogenase (LDH) assay. Additionally, more studies will be done comparing the data to E10 results, while using ELISA
and PCR analysis to detect the presence of the pro-fibrogenic cytokine, Transforming Growth Factor-b1 (TGF- b1) and the
anti-fibrogenic, Chemokine CXC- motif 10 (CXCL10). These studies will provide a better understanding of how surface
functionalization affects the toxicity of MWCNTs and their potential to cause disease in humans.
Research Grant: NSF International Collaborative on Nanotube Safety (ICONS) Grant 15-022.
Student Support: NIH- T-35 Interdisciplinary Biomedical Research Training Program and NIH Grant T35OD011070
97
An investigation into the epidemiology of B. odocoilei in Ontario ticks and cervids
Hannah Bagnall,Ellie Milnes, Dale A Smith, Nicole Nemeth
Department of Pathobiology (Bagnall, Smith, Nemeth), Ontario Veterinary College, University of Guelph, Guelph,
Ontario, Canada; Toronto Zoo (Milnes), Toronto, Ontario, Canada
Babesia odocoilei is an intraerythrocytic parasite transmitted by Ixodes ticks that can cause fatal infection in some cervid
species. While this parasite is endemic in the southern and eastern United States, it is now emerging as a cause of mortality
in captive and free-ranging cervids in Canada. In Ontario, cases have thus far been limited to captive reindeer and elk in a
zoological park. Given these recent diagnoses, the objective of this research is to describe the prevalence of B. odocoilei
in a variety of cervid species in Ontario, in both captive (e.g., zoos, game parks, commercial farms) and free-ranging environments, as well as in the arthropod vector (i.e., Ix. scapularis). Current sampling efforts include collecting ticks from
migratory birds at spring stopover sites at Long Point, Ontario to evaluate the potential for spread of infected ticks by birds.
Environmental sampling (tick dragging) was undertaken around Long Point and the Toronto Zoo to assess for the presence of B. odocoilei in native tick populations. In addition, spleens are being opportunistically collected from captive and
free-ranging cervids. A total of 42 ticks were found on 21/595 birds (3.5%) at Long Point between May 21-28, 2016. Tick
dragging yielded 27 ticks, predominately Dermacentor variabilis (64%). In addition, 9 spleens have been collected from
four cervid species. All tick and spleen samples are awaiting PCR for B. odocoilei. The results from this study will contribute to a better understanding of the potential impacts of B. odocoilei on wild and captive cervid populations in Ontario and
provide a baseline for future surveillance.
Research Grant: Toronto Zoo
Student Support: Andrea Leger Dunbar Summer Research Assistantship, Ontario Veterinary College
Volumetric analysis of the hippocampus and amygdala in California sea ions with domoic acid induced epilepsy
Anna L. Baker, Emily C. Abrams, Frances M. Gulland, Shawn P. Johnson, Paul S. Buckmaster
Utah State University - Washington State University, College of Veterinary Medicine, Pullman WA (Baker), Department
of Comparative Medicine, School of Medicine, Stanford University, Stanford, CA (Baker, Buckmaster, Abrams), Marine
Mammal Center, Sausalito, CA (Gulland, Johnson).
Temporal lobe epilepsy (TLE) is a common form of epilepsy in humans. It also occurs in California sea lions (Zalophus
californianus) exposed to the algal toxin domoic acid. Domoic acid is a glutamate receptor agonist that can cause status
epilepticus and permanent brain damage, resulting in TLE. Many human TLE patients have neuron loss in the hippocampus
and reduced hippocampal volume. In rodent models of TLE, hippocampal neuron loss and volume reduction is less severe.
However, rodent models show significant loss of amygdalar volume. In contrast, an MRI study of 241 human TLE patients
found no significant volume reduction of the amygdala. These differences in brain damage patterns may indicate differences
in pathogenesis. Consequently, potential treatments developed in rodent models might not translate successfully to humans.
The purpose of this study was to assess the degree of damage to the hippocampus and amygdala in sea lions with TLE induced by domoic acid toxicosis. Sea lions with TLE (n=6) and controls (n=4) were perfused with formaldehyde immediately
after euthanasia because of failed response to treatment and poor prognosis. Brains were sectioned coronally (40 mm), and
sections were stained with thionin. Series of sections are being analyzed using stereological methods and a Neurolucida system. Volumes of the amygdala and hippocampus are being estimated. Preliminary results suggest volume loss is more severe
in the hippocampus and less severe in the amygdala in epileptic sea lions, similar to human TLE patients. If confirmed, these
findings would suggest that sea lions may be a useful animal model for testing anti-epileptogenic treatments for human TLE.
Research Grant: NSF and NIH (NIEHS & NINDS)
98
Susan J. Baley, Julia B. Ponder, and Leslie C. Sharkey
The Raptor Center/Department of Veterinary Population Medicine (Ponder) and Department of Veterinary Clinical
Sciences (Sharkey), College of Veterinary Medicine, University of Minnesota, St. Paul, MN
Raptors are vulnerable to coagulopathies from secondary poisonings by consuming rodents poisoned with vitamin K anticoagulant rodenticides. Identification of rodenticide toxicity is difficult and a better diagnostic tool is needed to confirm
these toxicities in birds. A reliable point-of-care (POC) tool would be an invaluable diagnostic for raptor rehabilitators. The
specific aims of this study were: to determine the performance characteristics of a POC mechanical clot detection coagulometer for the measurement of prothrombin (PT) time and to assess the impact of the use of rabbit vs avian thromboplastin in
the measurement of PT time in raptors. We hypothesized that avian thromboplastin will give more precise results within the
reportable range of the analyzer as conspecific reagents have been previously reported to give less variable clotting times.
Rabbit thromboplastin was purchased from a commercial laboratory. Avian thromboplastin was made from the brains of
adult red-tailed hawks (Buteo jamaicensis)and great-horned owls (Bubo virginianus). Data collection is on-going. To date,
we have optimized the protocol to create avian thromboplastin and conducted multiple freeze/thaw trials to determine stability. Coefficient of variation (CV) is less than 8% in all trials. Initial data suggests that machine precision has been established
by measuring PT time using rabbit thromboplastin with quality controls from a commercial laboratory. CV was less than
5.4% for all three control levels. Future work includes troubleshooting consistently prolonged coagulation times with our
quality control material and rabbit thromboplastin sources.
Research Grant: Morris Animal Foundation
Kristen E Ballard, Ingeborg Langohr, Keith O Strother, Alma F Roy, Bonnie B Boudreaux
Louisiana Animal Disease Diagnostic Laboratory, Louisiana State University, Baton Rouge, LA (Ballard and Strother);
Department of Pathobiological Sciences (Langohr and Roy) and Department of Veterinary Clinical Sciences (Boudreaux),
School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA
Mast cell tumors (MCTs) are the most common skin tumor affecting dogs, therefore improved diagnosis and prognosis of
MCTs is crucial. Approximately 20% of MCTs are positive for a c-kit mutation that manifests as internal tandem duplications of exon 11 of c-kit gene. Tumors with the mutation exhibit a more aggressive behavior. Tyrosine kinase inhibitors
can be used specifically to target tumors with this mutation. Detection of a c-kit mutation requires tests to be completed on
histopathology samples. Detection of circulating c-kit gene and c-kit mutation in blood or urine (liquid biopsies) from dogs
with MCTs, however, would provide a less invasive technique to use for diagnostic, therapeutic and prognostic purposes. In
this study, cutaneous MCTs were removed and blood and urine were collected from eleven dogs; liquid biopsies were also
collected from five healthy, control dogs. Qiagen QIAamp kits were used to extract DNA from tissue samples and liquid
biopsies, and gel-based polymerase chain reaction was completed for detection of c-kit mutation. c-kit gene was detected
in the plasma of all dogs, including the control group, indicating that the presence of circulating c-kit gene in plasma from
dogs with MCTs is not beneficial for diagnosis. The detection of c-kit gene in urine had variable results, therefore urine is not
reliable for detection of circulating c-kit gene. Two of the eleven dogs with MCTs were positive for the mutation in biopsy
samples, but c-kit mutation in the corresponding liquid biopsies was not detected. In conclusion, the detection of circulating
c-kit gene in plasma was successful, but with the methods used in this study, detection of the c-kit mutation in liquid biopsies
was not accomplished.
Research Grant: Louisiana Animal Disease Diagnostic Laboratory and LSU SVM Department of Pathobiological Sciences
Student Support: Kenneth F. Burns Trust
99
Effects of 17-a-estradiol treatment on stifle joint morphology in aging mice
Channing Bancroft, Cathy Carlson, Richard Miller, and Richard Loeser
University of Minnesota CVM, St. Paul, MN
17-a-estradiol (EST), a non-feminizing estrogen that is closely related to 17-b-estradiol, recently has been shown to increase longevity in a natural aging study in male mice. The objective of the present study was to determine whether this
treatment influenced tissues in diarthrodial joints that are affected by aging, and hypothesized that osteoarthritic lesions in
the knee joint would be reduced in treated mice compared with untreated controls. The left hind limbs from 12 control and
12 EST-treated male mice, with ages ranging from 24-42 months (avg. 32), were received by our laboratory, fixed in formalin. After decalcification in 10% ethylene diamine tetra-acetic acid (EDTA), the stifle joints were routinely processed and
embedded in paraffin. Midcoronal sections from each block were cut at 4 mm and were stained with hematoxylin and eosin
(H&E). The integrity of the articular cartilage at four sites, including medial tibial plateau (MTP), medial femoral condyle,
lateral tibial plateau, and lateral femoral condyle, was evaluated using the Articular Cartilage Structure score previously
developed by our lab. Measurements of thickness and area of articular cartilage, calcified cartilage, and subchondral bone
in the MTP in each section were taken using the OsteoMeasure bone histomorphometry system. In addition, other lesions
associated with the joint tissues were observed and recorded. These included chondrification and ossification of ligaments,
osteophytes, degenerative changes in patellae and menisci, and tumors involving the bone marrow. The relationship of these
lesions with treatment group and age of the mice will be evaluated to determine if EST has an effect on the development of
age-related osteoarthritis.
Student Support: National Institutes of Health, Award Number T35OD011118
Impact of Paternal and Maternal Preconception Stress on Offspring Neurodevelopment and Stress Response
Caitlin M. Banducci, Bridget M. Nugent, Jennifer C. Chan, Tracy L. Bale
Department of Biomedical Sciences, College of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA
Changes in the environment that alter the homeostasis of an animal can impact neurodevelopmental programming of its
offspring. It is well established that the developing brain is sensitive to environmental factors, ranging from obesity to psychological stress during pregnancy such that negative psychosocial experiences in the mother can increase risk of schizophrenia, autism, anxiety disorders, and depression in her children. Although the effects of stress during pregnancy are well
established, the impact of lifetime stress, prior to conception, is less understood. Recent studies in the Bale lab suggest paternal stress exposures before conception can impact the offspring’s Hypothalamic-Pituitary-Adrenal stress axis (HPA) reactivity, suggesting that parental lifetime exposure to stress can have behavioral effects in offspring. HPA axis dysregulation
is common in neuropsychiatric disorders and often associated with behavioral endophenotypes similar to schizophrenia and
autism in animal models. The Bale lab has recently uncovered a mechanism whereby paternal stress before conception leads
to altered offspring neurodevelopment in mice. However, offspring outcomes following maternal stress prior to pregnancy
have not been thoroughly investigated. This research adds novel information to the Bale lab’s findings that parental stress
programs the developing brain by investigating the contributions of preconception stress in males and females on offspring
stress axis sensitivity and behavioral phenotypes. I hypothesize that like paternal stress, maternal preconception stress can
similarly reprogram offspring HPA reactivity and behavior by altering HPA axis programming and neurodevelopmental
gene expression.
Research Grant: R37MH108286, R27R33MH104184, PSOMH099910
Student Support: NIH Merial Veterinary Scholarship Program
100
Prevalence and pathology of Trypanosoma cruzi in coyotes and raccoons in a disease-endemic region of Texas
Rosa Banuelos, Carolyn Hodo, Erin Edwards, and Sarah Hamer
College of Veterinary Medicine, Texas A&M University, College Station, TX and Texas Department of State Health
Services, Uvalde, TX
Chagas disease is a vector-borne zoonotic disease caused by the protozoan parasite Trypanosoma cruzi. Transmitted by
blood-sucking insects of the genus Triatoma, Chagas disease is a major public health threat in Central and South America
and is increasingly recognized in the southern United States. Infected primates, humans, and dogs can develop chronic cardiac disease, but pathology in wildlife reservoirs is largely unknown. A limited number of wildlife studies in southern states
have revealed that up to 14% of coyotes are seropositive, and experimental infections have shown myocarditis in a small
proportion of infected raccoons. We hypothesized that wild populations of coyotes and raccoons in central Texas would be
infected with T. cruzi and show cardiac lesions similar to those found in infected dogs. Heart tissue and blood were collected
from 97 hunter-harvested coyotes and 24 raccoons from central Texas. Hearts were examined grossly and histologically.
Blood and heart samples were subjected to PCR to detect T. cruzi DNA and determine strain type. While there were no gross
lesions detected in any of the hearts, 5 coyotes (5%) and 14 raccoons (58%) were PCR-positive for T. cruzi and 3 positive
coyotes examined histologically displayed lymphoplasmacytic inflammation characteristic of Chagas disease. All of the
positive coyotes were infected with strain type TcI, while the raccoons were infected with TcIV. We conclude that T. cruzi
can cause histologic lesions in the hearts of coyotes in a similar fashion as it does in domestic dogs, and that these lesions
may be severe enough to lead to heart disease. Additional histologic results are forthcoming.
Research Grant: Harry L. Willet Foundation
Student Support: National Institutes of Health
Evaluation of improvement in joint function from intra-articular injection of NV-08 in dogs with OA joint pain
Megan Barbar, Masataka Enomoto, Beth Case, and Duncan Lascelles
Comparative Pain Research Laboratory (Barbar, Enomoto, Case, Lascelles) and Center for Comparative Medicine and
Translational Research, Department of Clinical Sciences (Lascelles), College of Veterinary Medicine, North Carolina
State University, Raleigh, NC.
Osteoarthritis is the most common orthopedic disease of dogs. Currently, there are limited approved treatment options and
these are administered systemically. In dogs with OA a single joint is often affected more than others. However, there are
currently no approved intra-articular (IA) treatments. Tumor necrosis factor-a (TNF-a) is a potential target for IA treatment.
A human TNF receptor Fc (TNFR Fc) domain fusion protein (Etanercept) decreased pain scores in knee OA with no adverse
effects. NexvetTM has developed a caninized TNFR Fc-fusion protein (NV-08). The aim of our study is to assess the potential for NV-08 to provide positive therapeutic effects when administered IA to dogs with OA. We conducted a pilot proof
of principle study using objective outcome measures: kinetic measures of limb use (AMTI force plates) and accelerometry
(Actical accelerometers). Using a non-randomized and open label design, dogs (n=11) with a predominantly single-limb
lameness received NV-08 injected into the affected joint. Kinetic data were collected prior to (baseline) and at 14 and 28
days following treatment and accelerometry data were collected continuously. Kinetic variables (peak vertical force [PVF]
and vertical impulse [VI]), and mean activity counts per minute over each week of the study were calculated. Changes from
baseline were derived for these variables and tested using paired statistical tests, and changes were compared to historical
placebo data. Analysis of these data will provide the first evidence of whether an intra-articular anti-TNF-a fusion protein
may provide pain relief in dogs with OA and inform larger, placebo controlled studies.
Research Grant: Nexvet Biopharma PLC
Student Support: VSP Summer Scholars Program
101
Assessing key proinflammatory cytokines in equine peripheral blood mononuclear and monocytic cells, in vitro
Brian G. Barnett, Kelsey A. Hart, Robert M. Gogal, David J. Hurley, Brina Gorham, Natalie A. Norton,
and Robert J. Williams.
Department of Large Animal Medicine (Barnett, Hart, Gorham, Norton) and Department of Veterinary Biosciences and
Diagnostic Imaging (Gogal, Williams), University of Georgia College of Veterinary Medicine, Athens, GA
The immune system responds to stimuli through a complex series of physiological events involving cell-cell contact, cytokine secretion and signaling. Maintaining a homeostatic balance between different cytokines is crucial for optimal immunity. Thus, when dealing with critically ill horses, the ability to quantify and interpret equine cytokine profiles can be an
effective diagnostic tool for assessing proinflammatory events and in guiding therapeutic protocols. In the present study,
peripheral blood mononuclear cell (PBMC) and monocytic populations were enriched from equine peripheral blood and
cultured in complete media, media + LPS (enterotoxin), and media + ConA (T cell mitogen). Supernatants were collected
after 6, 18, and 24 hour and evaluated for production of TNF-a, IL-1b, IL-6, and IL-10 via both ELISAs and the Milliplex
MAP Multiplex Assay. Griess assays were performed to measure nitric oxide. Cytokine production in media alone varied
by horse in both the PBMC and monocytic fractions, but did not change over 24 hours. In the presence of LPS, the PBMC
TNF-a and IL-10 levels significantly increased and IL-1b was increased at 24 hours compared to media, whereas with the
monocytes, TNF-a and IL-1b levels varied by horse and IL-10 levels significantly increased. In the presence of ConA, the
PBMC TNF-a and IL-10 levels were increased and IL-1b levels were comparable to media, whereas with the monocytes,
the cytokine levels were comparable to control. The Griess assay was unremarkable and the Milliplex data are still pending.
Our preliminary results show that PBMC and monocytic fractions varied in cytokine secretion based on stimulant with LPS
yielding the most significant effects for both cell fractions.
Student Support: Merial Ltd, Veterinary Medical Experimental Station, UGA College of Veterinary Medicine
Novel in vitro model system of canine degenerative myelopathy using direct conversion of skin fibroblasts
Kristen Bartholomew, Shibi Likhite, Kathrin Meyer, Sarah Moore, and Brian Kaspar
Department of Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH
(Bartholomew, Moore); Center for Gene Therapy, Nationwide Children’s Hospital, Columbus, OH (Likhite, Meyer,
Kaspar)
Canine degenerative myelopathy (DM) is a multisystem, adult onset degenerative disease of the central and peripheral
nervous system. Amyotrophic lateral sclerosis (ALS) and DM have similar disease progression culminating in paralysis
and death, with no treatments available. Both diseases likely share similar pathogenic mechanisms involving non-cell autonomous toxicity exerted by mutant Superoxide Dismutase 1 (mtSOD1). To better understand these mechanisms, a direct
conversion method was utilized to convert skin fibroblasts from DM-affected and control dogs into CNS cell types to create an in vitro model system. Dog skin fibroblasts were transduced with four retroviral vectors containing reprogramming
factors causing conversion to induced neural progenitor cells (iNPCs). After conversion to iNPCs, further differentiation
to astrocytes, neurons, and oligodendrocytes was accomplished using media specific to each cell type, and conversion was
confirmed with PCR and IF. To establish the in vitro model system and to test the hypothesis that astrocytes promote neuronal degeneration in DM, skin-derived astrocytes were co-cultured with GFP-expressing wild-type mouse motor neurons.
We expect that skin fibroblasts from DM-affected dogs can be readily converted to iNPCs and i-astrocytes. We expect that
DM-derived astrocytes will affect the axonal growth and survival of motor neurons similar to ALS astrocytes from mice and
humans. The focus of this work is to establish a fast and reliable in vitro model system to study disease pathogenesis and test
potential therapeutic intervention. Additionally, establishment of a direct conversion protocol for canine fibroblasts will be
helpful in understanding other canine neurological disorders.
Research Grant: NIH funding from multiple grants
Student Support: NIH T35 Training Grant 5T35OD010977-08
102
Investigating IgG subtype responses in horses infected with different mutations of EHV-1
Rachel M. Baumgardner, Carine L. Holz, Margaret E. Wilson, Gisela Soboll-Hussey
Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, Michigan State University,
East Lansing, MI
Equine herpesvirus-1 (EHV-1) is an alphaerpesvirus that causes respiratory disease, but can also cause epidemic abortion
storms and equine herpes myoencephalopathy (EHM). The pathology and immune response to EHV-1 in developing EHM
is not completely understood. To understand the contribution of different viral proteins for the development of EHM and
induction of immunity, a N752 mutation of the polymerase gene was generated to change the neuropathogenic potential
of EHV-1, and a glycoprotein D (gD) mutant was made by replacing gD1 with gD of EHV-4, which does not cause EHM.
It has been shown that IgG subtypes IgGa and IgGb are associated with protective humoral immunity, while the subtype
IgG(T) blocks complement activation and opsonization. Based on this information, the hypothesis of our project is that
infection with the less neuropathogenic strains (N752 and gD4 mutations) will induce a higher IgGb and IgGa levels than
the wild-type strain. Twenty-five yearling horses were infected with either a wild type EHV-1 Ab4 (n=8), EHV-1 N752
mutation (n=9), or EHV-1 gD4 mutation (n=8). Serum samples were collected prior to challenge, days 7, 14, 22, 28, 46,
and 74 post-infection. ELISAs were run to determine levels of IgG subtypes in all samples against a standard curve. We
assessed each isotype titer concentration for each group using this standard curve; statistical analysis was done by ANOVA.
All groups showed increases in IgGa and IgGb subtype titers starting on day 22 post-infection. IgGa titers were highest in
group gD4, but IgGb titers were highest in group N752. N752 horses also had the highest IgG(T) titers. Our experiment
provides a gateway for further research into EHV-1 immune modulation and response.
Student Support: NIH Grant No T35OD016477
Pet carriage of Staphylococcus aureus and S. pseudintermedius in the households of children with asthma
Erin Beasley, Shanna Ludwig, Andrea Christ, Kathryn Dalton, Elizabeth Matsui, Meghan F. Davis
College of Veterinary Medicine, North Carolina State University (Beasley); Department of Environmental Health
Sciences, Bloomberg School of Public Health, Johns Hopkins University, Baltimore, MD (Beasley, Ludwig, Christ,
Dalton, Matsui, Davis); School of Medicine, Johns Hopkins University, Baltimore, MD (Matsui)
Staphylococcus pseudintermedius is prevalent in pets, particularly dogs, and S. aureus, more common in people, has
been documented in pets. Prior work identified the mouth and perineum as the single most sensitive site for recovery of
S. aureus and S. pseudintermedius, respectively, from mammalian pets exposed to a person with MRSA infection. However, such results may not be generalizable to all populations. Therefore, the objective of this study was to determine the
prevalence and anatomical site carriage of S. aureus and S. pseudintermedius in mammalian pets from households selected
on the basis of enrollment in a study of childhood asthma. Households were visited three times at a three-month interval.
During each visit, available and amenable pets were sampled via swab at the nares, mouth, inguinal skin, and perineum.
Swabs were subjected to bacterial culture using previously-described methods and all presumptive coagulase-positive
staphylococcal isolates were identified to species via multiplex PCR. Statistical analyses were conducted in Excel and Stata 13. Between 1/2015 and 5/2016, 55 pets, including 36 mammals, from 32 households were enrolled and sampled. The
prevalence of S. aureus at the first sampling point was: cats (71% of 17), dogs (47% of 19). The prevalence of S. pseudintermedius was: dogs (42%) and cats (18%). The inguinum was the most sensitive single site for recovery of S. aureus, and
the perineum or nares were the most sensitive sites for recovery of S. pseudintermedius. These results differ from prior
work among MRSA-exposed pets, suggesting that exposure to human MRSA may impact the site distribution for staphylococcal recovery from dogs and cats, with both practice and research implications.
Research Grant: Davis NIH K01OD019918, Matsui NIH R01ES023447, Matsui NIH K24AI114769
103
Hormonal effects on adenoma development in a rat model of human familial colon cancer
Jessica A. Becher, Susheel B. Busi, James M. Amos-Landgraf
North Carolina State University CVM, Raleigh, NC (Becher); University of Missouri CVM, Columbia, MO
(Busi, Amos-Landgraf)
Colon cancer affects men at an earlier and higher rate compared to women. The Pirc (polyposis in the rat colon) rat model
demonstrates this sex disparity. It has been found recently that female hormones do not play a role in protection from colonic
tumors; however, the mechanism of male and stress hormone action on intestinal cancer is not known. We have also shown
that changes in the microbiome can influence tumor development. To determine the relationship of the testosterone and
corticosterone on tumor development and the gut microbiome, we used two cohorts of Pirc rats. We hypothesized that male
rats with increased corticosterone levels and female rats with DHT (dihydrotesterone) implants would have a higher tumor
burden. Male Pirc rats were cohoused in pairs or cohoused with a female. Fecal and serum samples were collected prior to
cohousing, and two weeks and six months after cohousing. Serum samples were analyzed for corticosterone concentrations
and fecal samples were analyzed for microbial OTU (operational taxonomic unit) percentages. Colonic tumor counts were
obtained at six months of age. The second study group consisted of female Pirc rats that were ovariectomized at one month
of age. Ovariectomized rats received DHT or placebo implants (n=10/group). Fecal samples were collected pre- and one
month-post surgery for microbiome analysis. Cohoused males with increased tumor burden had a trend of higher mean
corticosterone levels pre-cohousing and had a significant decrease in corticosterone levels between the two week and the
six month time points. These results potentially indicate that increased corticosterone levels overtime result in greater tumor
multiplicities long term. This investigation is ongoing.
Research Grant: University of Missouri faculty development grant
Student Support: American Society of Laboratory Animal Practitioners, GlaxoSmithKine, and IDEXX-BioResearch
Carolyn Belling, Jeffrey Lorch, and Michael Bonds
University of Wisconsin School of Veterinary Medicine, Madison, WI (Belling), U.S Geological Survey - National
Wildlife Health Center (Bonds, Lorch)
Snake fungal disease (SFD) is an emerging skin disease affecting wild snakes in eastern North America. While relatively little is known about its effects on wild snake populations, infections can be fatal and imperiled populations of timber
rattlesnakes and massasaguas are particularly vulnerable to the impacts of SFD. Ophidiomyces ophiodiicola (O.o.) is
the causative agent of SFD, but the disease can be challenging to diagnose due to the difficulty of isolating O.o. from the
plethora of environmental fungi and bacteria found on snake skin. To develop more selective conditions for isolating O.o.
from diagnostic samples, we cultured affected tissues at different temperatures and found that 328C is ideal for inhibiting
many non-target microorganisms while still allowing for growth of O.o. However, temperature alone still did not prevent
all non-target fungi and bacteria from overgrowing O.o. in some diagnostic samples. To further refine selective conditions,
sequential efforts are now focused on identifying bacterial and fungal species most commonly isolated from snakes, and
testing the effects of different culture media additives, such as antibiotics, on their growth. This information will then be
used to develop media that inhibits growth of these non-target species, while having little to no effect on O.o., making it
easier and more reliable to isolate and identify O.o., and to further investigate SFD. Studies examining the effects of media
additives are underway and will be presented.
Research Grant: U.S. Geological Survey
Student Support: School of Veterinary Medicine, University of Wisconsin, Madison
104
Comparison of enterotomy leak pressure among fresh, cooled, and frozen/thawed swine jejunal segments
Rebecca Belter, F. A. (Tony) Mann, John Middleton
Department of Veterinary Medicine and Surgery, University of Missouri, Columbia, MO
Secure enterotomy closure is essential because intestinal leakage may be fatal. Cadaveric tissue is used to test enterotomy
closure techniques; however, postmortem autolysis alters tissue integrity and may influence results. To accurately mimic
in vivo conditions, previous studies have used intestines within 5 hours after euthanasia, but the dependability of intestine
beyond 5 hours after death has not been evaluated. The objective of this study was to determine if stored tissue could be used
to obtain dependable leak pressure data after enterotomy closure at different times post mortem. Segments were divided
into three treatment groups: fresh, cooled overnight, and frozen/thawed. We hypothesized that there would be no difference
in leak pressure among the three treatment groups. Jejunum was harvested from three pigs immediately upon euthanasia,
and 36 jejunal sections (8 cm) were assigned to each group. For testing, each segment was suspended and sealed, and 2-cm
enterotomies were made and closed with a simple continuous suture pattern. Fluid was infused into each segment and leak
pressure was measured using a digital transducer. The mean leak pressures (mmHg) were 68.3, 55.3, and 14.4 with standard
deviations of 23.7, 28.1, and 14.8 for fresh, cooled, and frozen segments, respectively (p-values <0.001). Using the Bonferroni method, leak pressure differed between fresh and frozen, and between cooled and frozen, but not between fresh and
cooled. These findings, in light of 23 mmHg maximal intrajejunal pressure during peristalsis (dogs), indicate that segments
cooled overnight may be used for intestinal leak pressure testing, whereas frozen/thawed specimens should not.
Student Support: Department of Veterinary Medicine and Surgery, University of Missouri CVM
Acute toxicity evaluation in rats of potential chemical warfare agent antidotes.
Joshua P. Bennett, Edward C. Meek, Shane W. Bennett, Ronald B. Pringle, Howard W. Chambers,
and Janice E. Chambers.
Department of Basic Sciences (J. Bennett, Meek, S. Bennett, Pringle, and J. Chambers), College of Veterinary Medicine,
Mississippi State University, Mississippi State, MS. Department of Biochemistry, Molecular Biology, Entomology, &
Plant Pathology (H. Chambers), Mississippi State University, Mississippi State, MS.
Novel substituted phenoxyalkyl pyridinium oximes (US patent 9,227,937 B2), previously shown to reactivate rat brain
cholinesterase inhibited by nerve agent surrogates and enhance survival and decrease time to cessation of seizure-like
behavior following lethal dosages of nerve agent surrogates, were tested for potential acute toxicities in rats. Rat cohorts
were treated intramuscularly with 292 mmol/kg of oxime (2x the human equivalent dosage) or vehicle (Multisol) control
and were weighed daily after administration. There was no statistical difference detected in brain cholinesterase activity
between treatment groups at times of 1 hour, 24 hours, and 2 weeks post-dosing. In vitro testing of these oximes at 2 different concentrations using rat brain homogenate revealed a range of 2-18% inhibition of cholinesterase activity. In addition,
there were statistical differences detected in weight gain among Oximes 20 and 55 at 2 weeks post-dosing, but no difference
among treatments was found after 24 hours. Also in the 2-week cohort, a statistical difference was observed among the
kidney weights and indices of Oximes 1, 15, and 20. Post-mortem evaluation of the 2-week cohort’s organs showed changes
in kidney size and color, in some, of the rats treated with Oximes 1, 15, and 20. No other gross pathological changes were
noticed among treatment groups, except for irritation at the injection site. Previous studies performed in our lab have shown
that these oximes have exceptional promise as a potential therapy to prevent nerve agent-induced lethality and attenuation
of seizure-like behavior; therefore, further preclinical studies on safety and efficacy are needed.
Research Grant: NIH U01 NS083430
Student Support: Mississippi State University College of Veterinary Medicine
105
Susceptibility of Amblyomma americanum, Dermacentor variabilis and Rhipicephalus sanguineus to acaricides
Kelsey L. Bentz, Jennifer E. Thomas, and Mason V. Reichard
Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK
The purpose of this study was to evaluate the susceptibility of Amblyomma americanum, Dermacentor variabilis, and
Rhipicephalus sanguineus to three common acaricides: fipronil, permethrin and diazinon. Acaricide resistance has been
documented in multiple species of ticks across the world, and a recent study found resistance in R. sanguineus from the
United States. Laboratory-reared ticks are used experimentally to test the efficacy of acaricides and therefore must also be
evaluated for emerging resistance. We hypothesized that ticks from the Tick Rearing Facility at Oklahoma State University
were susceptible to current acaricides. Amblyomma americanum, Dermacentor variabilis, and Rhipecephalus sanguineus,
three of the most common species of ticks in the United States, were obtained from the Tick Rearing Facility at Oklahoma
State University. Each tick species was tested against four different concentrations of fipronil, permethrin and diazinon using the larval packet test. Dose-response curves were evaluated to assess susceptibility of tick larvae to the differing doses
of each acaricide. We found that laboratory-reared A. americanum, D. variabilis and R. sanguineus were susceptible to
fipronil, permethrin and diazinon. The average mortality 6 95% confidence interval was 99.6% 6 0.3 when exposed to
acaricides, while ticks not treated (i.e., controls) had an average mortality of 8.2% 6 3.0. Future research incorporating a
wider variety of acaricides, susceptibility of lab reared ticks in comparison to field strains, and additional species of ticks
will be discussed.
Research Grant: We acknowledge the support of the Oklahoma State University Center for Veterinary Health Sciences and
the Summer Scholars Research Program.
Student Support: Unkown
The Effect of High Density Lipoproteins on Bovine Neutrophil Activation
Bridget N. Bickers, Cheryl Wong, Daryl Nydam, Franco Leal-Yepes, Sabine Mann, Erica Behling-Kelly
Cornell University CVM, Ithaca, NY
High density lipoproteins (HDL) are responsible for the transport of cholesterol from peripheral tissues to the liver. In addition to HDL’s role in lipid transport, this lipoprotein has been shown to have anti-oxidative and anti-inflammatory properties
in mice and humans. Previous studies performed by our research team have shown that there is a marked decrease in HDL
during the transition period in cows. This is important because the transition period is the time when dairy cows are at their
highest risk for the development of costly inflammatory diseases. Therefore, we hypothesized that the decrease in HDL
we documented during the transition period contributes to immune dysregulation and the high incidence of inflammatory
disease seen in cows during the transition period. To begin to test this hypothesis, we first sought to determine if bovine
HDL has an effect on neutrophil activation. Whole blood from dry cows was treated with either lipopolysaccharide (LPS)
as a positive control, HDL, or HDL and LPS in combination. Using flow cytometry, changes in the expression of the adhesion molecule CD11b, a marker for neutrophil activation, were quantified. Lipopolysaccharide caused a robust increase in
CD11b expression whereas HDL decreased the effect of LPS. Assays measuring reactive oxygen species (ROS) formation
are being validated and will be used to determine if HDL has an anti-oxidative effect on bovine neutrophils. We are also
evaluating HDL isolated from cows during different life stages to determine if bovine HDL function changes with the physiologic and metabolic state of the cow.
Research Grant: None (Pending USDA application)
106
Sympathetic neurotransmitter induced vasoconstrictions in high fat and control diet fed rats
Jamie R. Binns, James J. Galligan , and Hui Xu
Biology Department (Binns) Talladega College, Talladega, Al and Department of Pharmacology and Toxicology
(Galligan and Xu) College of Human Medicine, Michigan State University, East Lansing, MI
Hypertension is a commonly diagnosed cardiovascular disorder affecting Americans. Studies have shown links between
obesity and hypertension. As higher vascular reactivity to sympathetic neurotransmitters causes arterial constriction, increases vascular peripheral resistance, and blood pressure, I hypothesized that obesity may increase the arterial responses to
sympathetic neurotransmitters (norepinephrine, NE; and ATP), which also contributes to obesity associated hypertension.
Male Sprague-Dawley rats were fed on high fat diet (HFD, 60% kcal n=5) or normal diet (NFD; 10% kcal from fat n=5)
for 20-22 weeks after weaning (3 weeks of age). Compared with NFD rats, HFD rats were obese (836639g vs 624656g,
P<0.05), normotensive (MAP, 10861 mmHg vs 10762 mmHg, P>0.05, measured by radiotelemeter). NE and ATP-induced
arterial constrictions were determined in cannulated and pressurized mesenteric arterial (~300mm innerdiameter, 60 mmHg)
from HFD and NFD rats in vitro. Vessels were perfused with Kreb’s buffer. Drug-induced changes in arterial inner diameter
were recorded. Concentration response curves for NE and ATP were plotted and the EC50 values of each drug (the concentration of a drug that gives half-maximal response) were calculated using Origin software. NE-induced maximal constriction
and pD2 (EC50) were 89610% and 6.360.2 (n=4) in NFD rats; 94% and 6.8 (n=1) in HFD rat. ATP-induced maximal
constriction and pD2 (EC50) were 44% and 1.2 (n=1) in NFD rats; 72% and 3.8 (n=1) in HFD rat. My studies indicated that
obesity may not be necessarily with hypertension in rats. The conclusions of altered arterial reactivity to neurotransmitter
cannot be made from my current studies, due to the lower animal numbers in HFD rats.
Research Grant: NIH 2P01HL070687-11A1
Student Support: NIH R25HL103156
Col3’s roles in the fibrotic response: therapeutic strategies to engineer regenerative and tumor environments
Jonah C. Binstock, Becky K. Brisson, Chelsea M Burgwin, and Susan W. Volk
Department of Clinical Studies, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA
Over 6 million Americans suffer yearly from chronic wounds or pathologic scarring, with health care costs exceeding $35
billion. Pathologic scars cause functional impairment, discomfort, and disfigurement. Myofibroblasts are contractile cells
important in wound repair; however, these cells can increase scar tissue. The Volk lab has shown that myofibroblasts can
be affected by type III collagen(Col3) in wounds. Col3’s regulation of myofibroblasts could provide treatments for chronic
wounds and aberrant scarring. We postulate that Col3 regulates myofibroblasts via its triple helical domain, with a11b1
integrins, and via a cysteine rich(CR) domain in the N-propeptide, by TGFb suppression. To elucidate the role of Col3 in
myofibroblast regulation, we exploited an in vitro fibroblast-populated stressed collagen contraction assay that mimics the
wound bed. Initial results show significant reduction in gel contraction by Col3WT cells with increasing Col3 (100%Col1
gels=24% contraction vs 50%Col1:50%Col3=10% contraction; p<0.0001). To test the CR domain’s role in myofibroblast
activity, the CR peptide was added to Col1 gels seeded with Col3KO cells. CR peptide reduced contraction by ~30%
(p<0.05). Future studies will determine if these Col3 domains decrease gel contraction via inhibition of myofibroblasts in
an a11 integrin and an TGFb-dependent manner. Several methods to quantitate Col3 have been published, including immunohistochemistry, polarized Pico Sirius Red, Herovici staining and Second Harmonic Generation imaging. Since these
methods have limitations, our second aim was to find the most accurate way to measure Col3. Work is ongoing using tissues
from Col3 WT, HET, and KO mouse embryos to decide which method is superior.
Research Grant: This work was supported by grants from the Mari Lowe Center for Comparative Oncology and NIAMS
(K08AR053945), as well as the Jack Miller-Ebrahimi Foundation.
Student Support: Supported by NIH grant T35 OD-010919 and a grant from Merial.
107
Trypanosoma cruzi strain types infecting triatimone vectors and primates at two research facilities in Texas
Elise C. Birkner, Carolyn L. Hodo, Stanton B. Gray, Gregory K. Wilkerson, Mark Cottingham, Geraldine Fleurie,
Rosa M. Bañuelos, Sarah A. Hamer
Department of Veterinary Integrative Biosciences (Birkner, Hodo, Bañuelos, Hamer), College of Veterinary Medicine
and Biomedical Sciences, Texas A&M University, College Station, TX; MD Anderson’s Keeling Center for Comparative
Medicine and Research (Gray, Wilkerson), Bastrop, TX; SNBL USA (Cottingham, Fleurie), Alice, TX
1
Chagas disease is a vector-borne zoonotic infection of mammals caused by the protozoan parasite Trypanosoma cruzi and
transmitted by triatomine insects (kissing bugs). T. cruzi is a genotypically heterogeneous species that has been divided into
six discrete typing units (DTUs), TcI - TcVI, each with varying geographic and epidemiologic associations. At a number
of nonhuman primate (NHP) facilities across the southern US, where T. cruzi is endemic in vectors and wildlife, primates
have become naturally infected with negative consequences for primate health and for their use as models for biomedical research. Documentation of the specific mechanisms of transmission of T. cruzi to NHPs is lacking, and there are no reports on
how the distribution of strain types in NHPs compares with that of the kissing bug vectors present in the immediate vicinity.
Our hypothesis is that the relative abundance of parasite strain types in infected NHPs is representative of that found in local
kissing bugs. At two NHP facilities in Texas with a history of Chagas disease in NHPs, we conducted passive and active
vector surveillance, and received blood and cardiac tissue samples from infected NHPs. All PCR-positive samples will be
strain-typed and preliminary results from 34 primates reveal a nearly equal infection rate with TcI and TcIV. Similarly, of 5
T. cruzi-positive bugs collected from one facility thus far, 2 are TcI, 2 are TcIV and 1 is a potential mixed TcI/IV infection.
Blood meal analysis using PCR and sequencing will also be performed on all kissing bugs. These findings will shed light
on the epidemiology of T. cruzi at NHP facilities and will help develop recommendations for controlling transmission of the
disease in the future.
Research Grant: Harry L. Willet Foundation
Student Support: NIH 5T35OD010991-12
Brianne Bishop, Dr. Scott Poock, Dr. David Patterson
College of Veterinary Medicine (Bishop), MU CVM Extension (Poock), MU Extension, Department of Animal Sciences
(Patterson), University of Missouri, Columbia, MO
As the number of food animal veterinarians across the United States has slowly declined, a proportion of practicing bovine
veterinarians have begun to offer new specialized services. The trend toward focusing on herd health rather than individual
animal health has been a long-standing goal for many veterinarians, but recently veterinarians have been adding reproductive, nutritional, and production management services to those offered for clients. It is unknown what effect these services
have had on the profitability of veterinarians and their clients, and if these services have any effect on the number of practicing food animal veterinarians. We hypothesize that veterinarians have successfully incorporated these services into their
practices for the last 5 to 10 years and that this has had a positive impact on their clients as well. Surveys will be sent to food
animal veterinarians throughout the state of Missouri in order to characterize the services being offered to clients and any
effect this may have on their practice. Surveys will also be sent to clients referenced by these veterinarians who can attest to
the improvements in their herds due to services that their veterinarian provides. It is expected that a majority of veterinarians
and clinics offer specialized services, and that this improves the profitability of their practices. Additionally, information will
be collected regarding training and programs that veterinarians receive or participate in that expand the use and availability
of these services. Characterizing changes in services provided by food animal practitioners across Missouri will answer
questions regarding the impact of these changes on veterinarians and producers across the state.
Student Support: USDA National Institute of Food and Agriculture, Animal Health project 1008625
108
Pharmacodynamic assessment of a panel of immunosuppressant drugs on ex-vivo canine T-lymphocyte proliferation
Kaitlin A. Bishop, Hans Rindt, Laura A. Nafe and Carol R. Reinero
University of Missouri, CVM, Columbia MO
A lack of understanding of specific immune defects underlying canine immune-mediated diseases hampers optimal therapy.
Failure to tailor treatment to an individual’s immune abnormality can result in lack of efficacy, added expense, and adverse
effects. We hypothesized that a small volume whole blood flow cytometric assay would show dose-dependent suppression
of T lymphocyte proliferation in response to dexamethasone, cyclosporine, mycophenolic acid, and the active metabolite
of leflunomide (A77 1726) in healthy dogs. Whole blood was collected from 6 healthy pet dogs, incubated for 4 days with
or without the mitogens concanavalinA and lipopolysaccharide, and with increasing concentrations of immunosuppressant.
Samples were subsequently stained with viability dye, and antibodies against the pan- T lymphocyte marker CD5 and the
cell proliferation marker Ki67. Percentages of proliferating T lymphocytes were determined by flow cytometry and the 50%
inhibitory concentration (IC50 ) was calculated. Inhibition of T-lymphocyte proliferation by the panel of immunosuppressants was shown in a dose-dependent manner, with marked variability among dogs. The mean 6 SD IC50 was 394.86 871
mM for dexamethasone, 18.896 36.2 ng/mL for cyclosporine, 106.36157.7 nM for mycophenolic acid, and 3.7466 6.8
mM for A77 1726. Results support the use of this assay for detecting efficacy of individual immunosuppressants to diminish
T lymphocyte proliferation. In the future it may be applied to pet dogs with spontaneous immune-mediated disease to help
tailor individual treatment regimens.
Student Support: A gift from Zoetis Animal Health
Development and performance analysis of a multiplex assay to detect exposure to tick-borne diseases in dogs
Kelley E. Black , Melinda J. Wilkerson, Marta Lanza-Perea, Bhumika Sharma, Kathryn Gibson, Diana M. Stone,
Anushka George, Arathy D. S. Nair, Roman R. Ganta, Michael W. Sanderson
Department of Diagnostic Medicine/Pathobiology and Kansas State Veterinary Diagnostic Laboratory (Black,
Wilkerson, George, Nair, Ganta, Sanderson), Kansas State University, Manhattan, KS; bSmall Animal Medicine and
Surgery Department School of Veterinary Medicine (Lanza-Perea) and Department of Pathobiology (Sharma, Gibson,
Stone), St. George’s University Grenada, West Indies
The goal of this study was to develop a serologic, multiplex bead-based assay to detect species-specific exposures to
Ehrlichia canis, Anaplasma platys, and E. chaffeensis in dogs for the purposes of surveillance and public health interventions. Peptides from specific outer membrane proteins of P30 for E. canis, OMP1X of A. platys, and P28-19/P28-14 of E.
chaffeensis were coupled to magnetic beads and assay (xMAP platform) was optimized. In experimentally infected dogs,
the multiplex assay successfully detected antibodies for E. canis and E. chaffeensis, but not A. platys. In 104 Grenadian
dogs, the multiplex assay, IFA, and SNAP 4Dx tests detected A. platys antibodies as well as Ehrlichia spp.. Kappa test
and Bayesian analysis showed good agreement of the multiplex assay with the IFA and 4Dx tests for E. canis, but had
poor agreement in detecting A. platys. In conclusion, the multiplex peptide assay performed comparably well in detecting
the seropositive status of dogs to E. canis as other commercial assays; however, more work needs to be done to assess
performance in populations of dogs with exposures to various Ehrlichia spp.. Future investigations are being planned to
determine the reasons for low seroreactivity to A. platys, for which we hypothesize is due to Grenadian dogs having strain
differences in the OMP1X peptide. *Note: Part of this work has been accepted for publication at Journal of Veterinary
Diagnostic Investigation.
Research Grant: Windward Islands Research and Education Foundation (WINDREF #SRGI-14005) located in Grenada,
West Indies, and Kansas State University Veterinary Diagnostic Laboratory
Student Support: NIH T32 Training Grant T35OD010979
109
Variation in bronchial size during respiration and cough in healthy compared to malacic dogs
Carolyn Blank, L. Abbigail Granger, Patricia Queiroz-Williams, Mat Stewart, Michael Kearney, and Lorrie Gaschen
Department of Veterinary Clinical Sciences (Blank, Granger, Queiroz-Williams, Stewart, Gaschen) and Department of
Pathobiological Sciences (Kearney), School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA
Airway malacia is a common cause of cough in dogs with tracheal collapse being most commonly diagnosed. Malacia of
bronchi as a sole entity is diagnosed using bronchoscopy and criteria for its diagnosis using less invasive modalities (fluoroscopy or CT) are poorly defined. Previous studies focus on classifying collapse in coughing dogs, and there is a lack
of information on what normal bronchial behavior is during respiration and cough in healthy dogs. A grading scheme for
airway collapse has been established using bronchoscopy. The purpose of this study is to document the amount of bronchial
collapse in healthy dogs during phases of respiration and cough using fluoroscopy and CT. The diameters of mainstem bronchi of 47 healthy dogs were measured and compared using fluoroscopy under respiration and during cough. Using positive
pressure CT and end-expiratory CT, mainstem bronchial diameters of 16 healthy dogs were measured and compared at
each phase. Fluoroscopic and CT images of dogs having airway collapse confirmed bronchoscopically were evaluated and
compared to measurements from the healthy dogs to determine if a reasonable cutoff distinguishing normal from excessive
airway collapse can be reported. Results are pending, but we expect that the percentage of bronchial lumen reduction during
respiration will be as high as 24%, correlating with the amount of collapse seen in the trachea using CT, and that more collapse will be seen during cough. We hypothesize that the collapse in bronchomalacia patients will be significantly greater
than that in healthy dogs. We also predict that the current criteria used to determine excessive airway narrowing (50% collapse) will overlap with measurements seen in non-coughing dogs.
Christopher J Bloom, GuangJun Zhang
Department of Comparative Pathobiology, Purdue University, West Lafayette, IN
Background: Cancer is essentially a genetic/genomic disease, as there are various gene mutations in cancer cell genomes.
Only a small proportion of gene mutations, called cancer drivers, are responsible for cancer initiation and development.
Cancer driver genes are key for development of new, targeted cancer therapies and novel markers for diagnosis and prognosis. Using zebrafish-human comparative oncogenomics, we have identified mdm1 as a cancer driver gene. Approach: To
elucidate molecular mechanisms of this new cancer gene, we have created a new zebrafish model for mdm1 using CRISPR.
In this study we aim to investigate apoptosis in the mdm1 mutant fish embryos. UV irradiation was employed to induce
apoptosis through DNA damage. Apoptosis was compared between mdm1 mutant and wild-type fish embryos using acridine
orange staining. To better track apoptosis in vivo, a reporter transgenic fish line was created using the apoptosis biosensor,
LSSmOrange-mKate2 caspase-3. Results: Fewer apoptotic cells were found in the mutant embryos compared to wild-type
after UV-irradiation. In addition, the new reporter transgenic fish line reliably detects apoptosis in vivo. Conclusions: Mdm1
mutants may be less sensitive to UV-induced DNA damage. The LSSmOrange-mKate2 caspase-3 reporter transgenic fish
can be used for further investigation of the roles of mdm1 in DNA damage response and apoptosis in the future.
Research Grant: Hayward Foundation
110
Optimizing the broth environment for large volume growth of “Brachyspira hampsonii”
François Boire, Matthew Loewen, and John Harding
Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, SK
“Brachyspira hampsonii” is a novel causative agent of swine dysentery (SD), a disease characterized by mucohaemorrhagic
diarrhea and typhlocolitis. Most common in grower-finisher pigs, SD is associated with acute or chronic diarrhea, and a
decrease in weight gain and feed efficiency. Among the many Brachyspira species, our lab is exploring the pathogenesis of
“B. hampsonii” clade II strain 30446, a strain first identified in Saskatchewan, Canada. Our conventional broth anaerobic
culture growth methods are capable of achieving a concentration of 108 to 109 copies target DNA per millilitre using RTPCR testing. However, the environmental conditions of the media during the culture period are yet unknown. In addition,
the media environment and culture densities have not been replicated at volumes exceeding 1.0 L. Production of larger
volumes would allow for the efficient purification of protein produced by “B. hampsonii” strain 30446 required for future
studies, and enhance the development of potential vaccine prototypes. Using Brachyspira hyodysenteriae as a model, we
hypothesized that growth conditions include an anaerobic atmosphere of 90% N2 and 10% CO2 with a media pH similar to
swine colons (6.1-6.6). Preliminary data supports the atmospheric conditions with culture density similar to that of volumes
less than 1.0 L. In addition, pH decreased from 6.8 to 5.1 within 43 hours of incubation, potentially limiting growth of the
culture determined to have a final density of 1.35 x109 target DNA copies/mL. The effect of maintaining the pH above 6.1
on final culture density is underway.
Research Grant: Alberta Livestock and Meat Agency (ALMA); Agriculture and Agrifood Canada Agriculture Innovation
program (AIP)
Student Support: 2016 Merial Veterinary Scholars Program
Application of additive manufacturing technology and stem cell recruitment to improve osseointegration
Mary K. Bollman, Raphael A. Malbrue, Shengmin Guo, and Shaomian Yao
Department of Comparative Biomedical Sciences (Bollman and Yao), Department of Laboratory Animal Medicine
(Malbrue), School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA and Department of Mechanical
and Industrial Engineering (Guo), Louisiana State University, Baton Rouge, LA
Osseointegration is the structural and functional connection between bone and a load-carrying implant, and establishment of
strong osseointegration is the key factor for successful implant surgery. The common problems causing failure of implant
surgeries include slow establishment of osseointegration, partial osseointegration and loss of osseointegration overtime. Selective laser melting (SLM) 3D printing is an additive manufacturing technology that is capable to produce patient-specific
metal implants. With SLM, implants designed to have a interconnecting porous structure were fabricated. The implants were
coated with a biodegradable polymer, i.e., Polycaprolactone (PCL)/gelatin and a stem cell chemoattractant (stromal-cell
derived factor-1 alpha or SDF-1a). We hypothesize that the implants with porous structure coated with PCL/gelatin can
reserve SDF-1a for sustained release to recruit endogenous stem cells to the implants, in turn, can promote osteogenesis to
establish strong osseointegration. To test the hypothesis, the implants were placed in medium and the medium were collected at different times. The SDF-1a released to the medium was determined by ELISA. An in vivo experiment was conducted
by surgically inserting the implants into the skulls of rabbits. The animals were sacrificed around 6 weeks post-surgery, and
osseointegration of the implants was analyzed by reverse torque test, scanning electron microscope analyses and histology
study. Generally, the implants coated with SDF-1a or PCL/gelatin appeared to have better osseointegration than non-coating control. Maximal osseointegration was seen on implant coated with 200ng SDF-1a. The relationship between SDF-1a
releasing and osseointegration is discussed.
Research Grant: LSU Biomedical Collaborative Research Program (LBCRP) grant
111
Effect of intranasal cannabinoid administration on an experimentally induced rhinosinusitis mouse model
Glenn E. Brado and Colleen C. Hegg
Department of Pharmacology and Toxicology, College of Veterinary Medicine, Michigan State University,
East Lansing, MI
Chronic rhinosinusitis significantly reduces the quality of life in both humans and animals. Prolonged inflammation of the
nasal epithelium causes hyposmia, which interferes with taste and appetite, leading to health problems such as malnutrition
and unintended weight loss. Activation of the CB2 cannabinoid receptor, found on most immune cells, induces anti-inflammatory processes. Targeting CB2 receptors in the nasal cavity with selective agonists may alleviate inflammation in
rhinosinusitis. We hypothesize that CB2 selective cannabinoids will significantly decrease vascular permeability, mucus
production and neutrophilic infiltration in an experimentally induced rhinosinusitis mouse model. Twenty-four 6-8 week old
Swiss-Webster mice were randomly separated into four treatment groups (n=6). Each mouse intranasally aspirated lipopolysaccharide (2 mg/kg LPS) or vehicle (0.9% saline) for three days followed by a selective CB2 receptor agonist, JWH 133 (5
mg/kg), or vehicle (5% ethanol/5% cremaphor) for 5 days. Evans blue extravasation was performed to assess and compare
vascular permeability between treatment groups using spectrophotometry. Tissue was fixed (4% paraformaldehyde) and 20
micron frozen tissue sections of the nasal cavity (T1-T3) were collected. Histochemistry and immunohistochemistry was
completed to evaluate the sections for mucus production and neutrophilic infiltration respectively. We predict that the JWH
133 treatment will significantly decrease vascular permeability, mucus production, and neutrophilic infiltration in the nasal
cavity when compared to the LPS-only treatment. These findings will contribute to the development of therapeutic strategies
for treatment of rhinosinusitis.
Research Grant: Michigan State University Institutional Funds
Cassidy Briggs, Julian Trachsel, Heather K. Allen, Crystal L. Loving
Iowa State University College of Veterinary Medicine Summer Scholars Program, Ames, IA (Briggs) FSEP USDA-ARS
National Animal Disease Center, Ames, IA (Trachsel, Allen, Loving) Interdepartmental Microbiology Doctoral Training
Program, Iowa State University, Ames, IA (Trachsel)
With recent limitations to antibiotic use in livestock species, alternatives are sought to maintain swine health. Dietary resistant starch (RS), such as raw potato starch, is known to modulate microbial populations, increase short chain fatty acids
(SCFA), and modify mucosal gene expression in the swine cecum and colon. SCFA’s, notably butyrate as the preferred energy source for colonocytes, are thought to improve intestinal health, but the exact mechanisms are not fully defined. In order
to better describe the effects of dietary RS on swine intestinal health a study was completed to determine in vivo changes in
host immune status after feeding a RS diet for 3 weeks post-weaning. Two groups of piglets were fed either a standard diet,
or the same diet amended with 5% raw potato starch and then euthanized for collection of samples from the large intestine.
Cecal contents from the RS group had increased butyrate levels compared to the control group. Lymphocyte populations
in cecal tissue were evaluated with flow cytometry and immunohistochemical (IHC) staining. Flow cytometry revealed no
differences in the percentage of CD3+ cells, but piglets fed RS had lower ratio of CD8a+ to CD4+ T cell populations, and
a greater percentage of regulatory T cells (CD25+/FoxP3+) within the CD8a+ T cell population. IHC staining substantiated
no overall difference in CD3+ cells, and there was no difference in the number of IgA-secreting cells. Changes in gene expression in cecal mucosal scrapings were evaluated with RT-qPCR. RS piglets had increased expression of MUC2 and IL-6
mRNA. Overall, these data show some modulation of cecal immune status after feeding RS and provide additional data on
the effects of RS on swine enteric immunity.
Research Grant: Food Safety and Enteric Pathogens Research Unit, USDA-ARS, National Animal Disease Center,
Ames, IA
Student Support: Merial Limited & Iowa State University College of Veterinary Medicine
112
Evaluation of Plasmodium knowlesi in vitro susceptibility to methotrexate
Jacqueline K. Brockhurst, Roberto R. Moraes Barros, Juliana M. Sa, Thomas E. Wellems
Malaria Genetics Section, Laboratory of Malaria and Vector Research, National Institute of Allergy and Infectious
Disease, NIH, Rockville, MD
Plasmodium knowlesi is an emerging zoonotic malaria parasite found naturally in macaques and capable of causing severe
disease in humans. Anti-folate drugs are commonly used to control P. knowlesi in clinical and laboratory settings. Methotrexate (MTX), an anti-folate drug, has been shown to be effective at suppressing blood parasite levels in a baboon model
of knowlesi malaria and is active in vitro against even drug-resistant strains of Plasmodium falciparum. To investigate
the potential usefulness of methotrexate susceptibility as a selectable marker in future P. knowlesi transfection studies, we
determined the half maximal inhibitory concentration (IC50) in the common laboratory “H strain.” Parasites maintained
in culture were plated on 96 well plates and exposed to serially diluted drug concentrations for 36 hrs. Two strains of P.
falciparum were tested as controls. Parasite growth was determined using SYBR Green I DNA dye. The average IC50 for
methotrexate in the H strain of P. knowlesi was found to be 431.2 nM, which is ten-fold higher than values determined
for the P. falciparum strains. The high concentration needed to inhibit parasite growth in vitro suggests that Plasmodium
knowlesi H strain has some natural high tolerance to methotrexate and more work needs to be done to characterize the safety
and efficacy of the drug in vitro and primate models.
Research Grant: Funding provided in part by the Division of Intramural Research/NIAID/NIH
Student Support: CBSTP Summer Internship Program in Biomedical Research for Veterinary Medical Students
Emily B. Bucak, Erika B. Menezes, Fagner C.P. Santos, Sonya M. Baird, Shien Lu, Frank T. Austin, Abdullah Kaya,
Arlindo A. Moura, Erdogan Memili
College of Vet Med (Bucak), Depts. of Anim & Dairy Sci (Bucak, Menezes, Santos, Memili), Biochem, Mol Bio,
Ent & Plant Path (Baird, Lu), Pathobiology & Pop Med (Austin), Mississippi State Univ, Mississippi State, MS, USA;
Dept. of Repro & AI (Kaya), Selcuk Univ, Konya, Turkey; Dept of Anim Sci (Moura), UFC, Fortaleza, Brazil
Bull fertility is crucial for sustainable and efficient reproduction of cattle. Despite producing large amounts of sperm with
normal motility and morphology, some bulls have low fertility data. Molecular, cellular and physiological mechanisms
causing low fertility are elusive. The objectives of this study were to determine the cellular parameters of cryopreserved
sperm from 10 commercial Holstein bulls with different fertility data and ascertain bacterial presence in whole semen samples. Sperm parameters were characterized using Computer Assisted Sperm Analysis (CASA), light microscopy, and flow
cytometry. Sperm viability, morphology, velocity parameters, as well as cell membrane, DNA, and acrosome integrity were
analyzed. Bacterial presence was evaluated using biochemical tests and Polymerase Chain Reaction (PCR). CASA analysis showed a statistically significant difference between high (HF) and low fertility (LF) bull samples in average pathway
velocity, amplitude of lateral head displacement, and straightness. Both light microscopy and flow cytometry analyses did
not show any statistical difference in sperm parameters between high and low fertility bulls. Results of the biochemical
tests showed that only two bull samples have bacterial presence; a HF bull with Pseudomonas aeruginosa/putida (100%),
and a LF bull with Comamonas testosteroni (75.48%), Pseudomonas alcaligenes (11.45%), and Pseudomonas flourescens
(8.67%). PCR analysis of 16S ribosomal subunit DNA and gel electrophoresis showed that all semen samples had detectable
bacterial DNA. These findings are significant because they help advancement in fundamental animal science and biotechnology.
Research Grant: MAFES, USDA-ARS Biophotonics Initiative #58-6402-3-018, Brazilian CNPq and CAPES
Student Support: Merial Veterinary Scholars and MSU College of Veterinary Medicine
113
Identification of biomarkers for xenoestrogenic exposure using a prepubertal porcine model
Jordan L. Burns, David Ledoux, Kim Dae Young, Tim J. Evans
Department of Veterinary Pathobiology and Veterinary Medical Diagnostic Laboratory, College of Veterinary Medicine
(Burns, Kim, and Evans), Division of Animal Sciences, College of Agriculture, Forestry, and Natural Resources (Ledoux),
University of Missouri, Columbia, Missouri
Exogenous compounds (xenobiotics) with estrogenic activity are referred to as xenoestrogens, and many naturally-occurring and synthesized chemicals have been identified as having xenoestrogenic potential. There are increasing societal
concerns about the risks posed to humans and domestic animals by low-level exposures to xenoestrogens, as well as other
chemicals capable of interfering with normal hormonal function. Zearalenone (ZEN), a common mycotoxin frequently
detected in cereal grains consumed by humans and domestic animals, is an example of a naturally-occurring xenoestrogen.
Since swine share many anatomical and physiological similarities with humans, as well as other domestic animals, porcine
models are gaining in popularity for use in comparative biomedical research. The well-recognized susceptibility of prepubertal gilts to ZEN-induced hyperestrogenism and their physiological and anatomical similarities to juveniles of numerous
other species, underscore the suitability of a prepubertal porcine model to identify potential antemortem and postmortem
biomarkers for xenoestrogenic exposures. It was hypothesized that morphometric biomarkers for xenoestrogenic exposure
will be identified or refined in prepubertal gilts following 21 days of exposure to 0, 0.5, or 1 mg of zearalenone/kg of diet.
Vulvar width and length were measured during the course of ZEN exposure, and sections of the vagina, uterine body, uterine horns, uterine tubes, and ovaries, were collected and fixed in 10% neutral-buffered formalin, and stained for histologic
examination. Treatment group means for vulvar area calculated several different ways and various measurements performed
on sections of specific tissues will be compared using ANOVA.
Research Grant: Ledoux Extramural Funding
Student Support: University of Missouri College of Veterinary Medicine
Sydney D Byerley, Kari A. Verner, Russell P. Main
College of Veterinary Medicine (Byerley) and Weldon School of Biomedical Engineering (Verner, Main) and Department
of Basic Medical Sciences, College of Veterinary Medicine (Main), Purdue University, West Lafayette, IN
Osteocytes are the primary cells composing mature, healthy bone tissue. Osteocytes play key roles in bone modeling and
remodeling throughout the body. The cytoplasmic processes of osteocytes connect and communicate with other osteocytes
in the bone to allow for response to mechanical stimulation or injury. The purpose of this study is to quantify and compare
osteocyte densities and the number of primary canaliculi per osteocyte in the femoral midshafts of representative species of
birds, mammals, and reptiles. Femoral bone samples were harvested from multiple guinea fowl, mice, and monitor lizards.
Each bone was fixed, dehydrated, and stained with fluorescein isothiocyanate isomer I (FITC). Bones were embedded,
sectioned, and placed on microscope slides. Images were taken with an inverted confocal microscope and analyzed using
digital image software. The number of osteocytes and primary canaliculi will be counted and compared between the three
species examined. Because bone is an ancient tissue, one would expect homogeneous osteocyte density and canaliculi
counts between species. Differences in these baseline values could relate to species differences in the skeletal response to
mechanical load and injury. Future research projects will study the response to mechanical stimulation when taking osteocyte density and primary canaliculi numbers into account.
Research Grant: National Science Foundation
114
Improving adjunctive treatment of drug resistant diabetic foot infections
Troy A. Cabral, Siana Hoffman, David F. Ackart, Alexandra Todd and Randall Basaraba
Department of Microbiology, Immunology and Pathology, Colorado State University, Fort Collins, Colorado
Diabetic foot infections (DFIs) with multidrug resistant bacteria (MDR) are a common and serious complication of uncontrolled diabetes. Diabetes is the number one cause of distal limb amputations in humans with recalcitrant DFIs. Among the
common causes of DFIs are biofilm forming strains of drug susceptible and multi-drug resistant Staphylococcus aureus
(MRSA). Currently, antibiotic treatment for DFIs are prolonged or ineffective due to genotypic resistance or through biofilm
formation. Newly isolated 2-aminoimidazole (2AI) compounds have recently shown effectiveness at reducing phenotypic
and genotypic resistance by disrupting biofilms and disabling enzymatic mechanisms that are essential to bacterial resistance. Such small molecules have the potential to significantly improve current antibiotic treatment responses when used as
an adjunctive therapy to disrupt MRSA biofilms. We hypothesize that an effective treatment for diabetic MRSA infections
may be established by combining these compounds with clinically appropriate antibiotics. Our in vitro model involves infecting a collagen-based substrate for MRSA biofilm formation to screen drug combinations in vitro prior to in vivo testing
in animals. The effectiveness of treatment is determined by counting CFUs or through standard MIC assays. This study
showed that combining 2AI with antibiotics led to significant decreases in logCFU/mL counts (1 to 5 logCFU/mL reduction
when combined with Vancomycin, Sulfamethoxazole/trimethoprim and Daptomycin), as well as substantial drops in MICs
(4- to 8-fold decrease in Vancomycin and Daptomycin MICs). These data suggest that 2AI compounds may have value in
the treatment of recalcitrant DFIs in diabetic patients.
Student Support: Merial, Ltd.
Valeria L. Caceres, Anqi Fu, Chi-chung Hui, and Joan S. Jorgensen
Department of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin - Madison, Madison,
WI (Caceres, Fu, Jorgensen), Department of Molecular Genetics, University of Toronto, Toronto, ON, Canada (Hui)
Premature ovarian insufficiency (POI) affects one percent of women. Its negative effects on women’s fertility and health are
of major concern; however, its specific cause remains unknown. The Jorgensen lab discovered that global disruption of Irx3
and Irx5 (Irx3/5) in mice caused defects in ovarian follicle morphology and massive oocyte death. Follicles had granulosa
cells that were misshaped and poorly organized. Interactions between the oocyte and surrounding granulosa cells were severely disrupted. These data suggest that Irx3/5 promote cell-cell interactions within the follicle to maintain oocyte and follicle survival. Notably Irx3/5 are expressed in both the oocyte and supporting granulosa cells at distinct times during follicle
formation. My hypothesis is that elimination of Irx3/5 specifically in pre-granulosa cells will be sufficient to disrupt follicle
integrity. Sf1Cre mice were bred to Irx3f/D;Irx5G/G mice to generate offspring where Irx3 is eliminated only in pre-granulosa
cells in the context of a global deletion of Irx5 (Sf1Cre;Irx3/5 sFD). Adult ovaries from Sf1Cre;Irx3/5 sFD, and two controls,
wild type and No Cre;Irx3/5 sFD, were embedded together as a tissue microarray. Every fifth section was stained with H&E
and imaged. Structures including primordial, primary, secondary, antral and atretic follicles and corpora lutea will be counted in each section for each ovary. The counting will be done blindly to ensure no bias. We expect mutant ovaries to have
more atretic follicles and fewer corpora lutea. This study is important to determine whether Irx3/5 expression in the ovarian
somatic cells contribute to the healthy development of ovarian follicles and will give us new insight into the cause of POI.
Research Grant: Research Grant: NIH-R01HD075079 (JSJ)
Student Support: NIH, T35OD011078 (PI: Bjorling) Short-term Research Training for Vet Students in WI
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Linking canine distemper virus susceptibility to SLAM sequences in wildlife species
Bridgette R. Cahill, Karen L. Bailey and Melinda A. Brindley
Faculty of Science, School of Veterinary Science, The University of Queensland, Gatton, QLD (Cahill),
Department of Infectious Diseases (Bailey), Department of Population Health, Center for Vaccines and Immunology,
College of Veterinary Medicine, The University of Georgia, Athens, GA (Brindley)
Canine distemper virus (CDV) is a morbillivirus with an unusual broad host range. Unlike measles that infects humans,
rinderpest that infects cattle, and peste des petits that infects sheep and goats, CDV infects a wide range of carnivore species,
including dogs, skunks, raccoons, bears, and large felids. It causes a multisystemic disease primarily affecting the gastrointestinal tract, respiratory and neurological systems, and progresses to fatal disease in some species. The viral receptor
binding protein (H) interacts with two different cellular receptors during entry. First it binds with the signalling lymphocyte
activating molecule (SLAM) receptor expressed on host monocyte cells and then uses nectin-4 during later infection for
entry into epithelial cells. SLAM is known to have high diversity among carnivore species, whereas nectin-4 is more highly
conserved. Sequence analysis of the SLAM binding H protein in infected animals has classified nine different CDV lineages
worldwide. The different groups correlated with geographic distribution of the samples. We hypothesized that the carnivore
species that are susceptible for CDV will contain a conserved receptor binding site, whereas those carnivore species that do
not get infected with CDV will contain altered amino acid residues in the SLAM protein, preventing CDV interaction. To
begin to address this hypothesis, we sequenced SLAM genes from the striped skunk, raccoon, northern river otter and gray
fox from North America and developed a phylogenetic tree alongside known SLAM sequences, extracted from GenBank
and DNA Data Bank of Japan, to analyze genetic relationships.
Generation of a recombinant PIV5 virus expressing Zika antigen for vaccine development.
Robert Trey Callahan, Zhuo Li, and Biao He.
Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA.
Zika virus, an enveloped, positive-sense, ssRNA virus in the Flavivirus family, is currently an international public health
concern that the Americas, parts of Oceania, and many of the Pacific Islands are facing today. The World Health Organization (WHO) declared Zika virus a Public Health Emergency of International Concern and the Centers for Disease Control
and Prevention (CDC) activated its Emergency Operations Center to Level 1, the highest level of action to fight the virus. A
mosquito-borne virus, Zika causes a mild disease that includes fever and rash, as well as some joint pain and conjunctivitis
in healthy adults. If pregnant women are infected, Zika can cause severe birth defects in the fetus including microcephaly.
With the virus rapidly continuing to spread and cases of Zika-related microcephaly continuing to rise, the need for a vaccine is critical. Parainfluenza virus 5 (PIV5), commonly known as canine parainfluenza virus, is a promising vaccine vector
in humans because the virus has shown to be an efficacious vector for vaccine development, as well as it does not cause
disease in infected humans. Previously, our lab has successfully generated vaccine candidates for influenza, HIV, and the
rabies virus using PIV5 as a vector. In this work, our goal is to generate a recombinant PIV5 virus expressing Zika antigens
and test them as potential vaccine candidates. The Zika envelope (E) protein and membrane (M) protein will be cloned into
a PIV5 lacking SH (PIV5DSH) backbone using molecular cloning techniques. The recombinant virus will be verified by
sequencing. The recovered virus will then be used to immunize mice and efficacy of the vaccine candidates will be tested.
Research Grant: Fred C. Davison Endowment
Student Support: NIH Office of Research Infrastructure Programs, Grant Number 4T35OD010433-10
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The role of Hus1 impairment in non-small cell lung cancer tumorigenesis
Jasmine G. Calle, Tina Abratte, and Kelly R. Hume
Cornell University College of Veterinary Medicine, Ithaca, NY
Lung cancer is the leading cause of cancer-related deaths in the United States. Non-small cell lung cancer (NSCLC) is the
most common form of lung cancer. As K-RAS is a gene implicated in NSCLC, mouse models with the activating mutation
K-rasG12D are frequently used to study this cancer. Our research explores impairment of the DNA damage response gene
Hus1 as a mechanism to halt lung cancer. HUS1 is part of a heterotrimeric Rad9-Rad1-Hus1 complex that plays key roles in
checkpoint signaling and DNA repair. Previous studies have suggested that there is a requirement for Hus1 following K-ras
oncogene activation as tumorigenesis progresses. Therefore, targeting Hus1 may offer insights for treatment of NSCLC,
though partial inactivation in mouse models is necessary as complete inactivation leads to early embryonic death. Due to its
role in the DNA damage response, we hypothesize that mice with partial Hus1 impairment will be more sensitive to DNA
damage, which may affect tumor growth. To achieve this, we use mice with partial Hus1 impairment (Hus1neo/D1n) that are
afflicted with NSCLC via oncogenic K-ras. Mice receive doxycycline to activate tumor formation. After 6 months of oncogene activation, tumor size is measured. This study may reveal the potential of impairing Hus1 as a treatment strategy for
NSCLC.
Research Grant: American Lung Association/American Lung Association of New York (RG-192135-N;KRH)
Student Support: Office of Graduate Education - Cornell University College of Veterinary Medicine
Plasmin induces up regulation of matrix metalloproteinase-13 in murine bone marrow derived macrophages
Jason Cardwell and Bryan Copple
Department of Pharmacology and Toxicology Michigan State University East Lansing, MI
Liver fibrosis is a disease in which excess connective tissue is deposited in the liver. It has many causes, including hepatitis
B, hepatitis C and alcohol. The agents produce chronic liver injury which transforms hepatic stellate cells into myofibroblasts that deposit excess collagen in the liver. Liver fibrosis may advance into cirrhosis, which can be fatal. Treatments for
liver fibrosis include liver transplant, antivirals, and cessation of alcohol consumption. Liver fibrosis has been shown to be
reversible, even in advanced stages, through an increase in production of matrix metalloproteinases by macrophages. It is
unclear what stimulates macrophages to produce MMPs during reversal of fibrosis. Previous studies from our laboratory
suggest that plasmin is critical for macrophage production of MMPs during liver injury. Therefore, these studies seek to
test the hypothesis that plasmin increases expression of MMPs in bone marrow macrophages of mice by activating protease-activated receptor-1 (PAR-1). To test this hypothesis, wild type mice and PAR-1 knockout mice will be euthanized and
macrophages will be derived from their bone marrow. These two groups will be further divided into no treatment groups,
and treatment groups, with the treatment group within each genotype receiving plasmin. We saw an increase in MMP13
levels in WT and PAR-1 KO mice treated with plasmin. PAR-1 KO mice had twice the amount of MMP13 as WT mice.
In the future we hope that the successful identification of the receptor that plasmin acts on will allow researchers to design
anti-fibrotic drugs to combat liver fibrosis.
Research Grant: Michigan State University
Student Support: MSU College of Veterinary Medicine Biomedical Research Program
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The effects of metformin and resveratrol on canine hemangiosarcoma cell lines
Ariel Carlson, Karen Hererra, Beshay Zordoky, and Marianne Grant
College of Veterinary Medicine, University of Minnesota, St. Paul, MN (Carlson), St. Catherine University, St. Paul, MN
(Hererra), and Department of Experimental and Clinical Pharmacology, University of Minnesota, Minneapolis, MN
(Zordoky, Grant)
Canine hemangiosarcoma (HSA) is an aggressive malignant tumor of vascular origin oftentimes with poor patient prognosis. Metformin, a common human anti-diabetic drug, and resveratrol, a natural plant polyphenol, have been shown to
possess strong anti-proliferative and/or pro-apoptotic effects in several human cancer cell lines and appear to be non-toxic
in dogs. The effects of both metformin and resveratrol are thought to be mediated by activating the AMP-activated protein
kinase (AMPK). We hypothesized that metformin and resveratrol have growth inhibitory effects on canine HSA cell lines.
HSA cell lines Frog and DD-1 were incubated with increasing concentrations of metformin and resveratrol with or without
doxorubicin, a common cytotoxic agent. The growth inhibitory effects of either metformin or resveratrol were assessed
by MTT assay. Western blotting was used to assess the activation of AMPK and the expression of several pro-apoptotic
proteins. In contrast to human cell line studies, metformin had a minimal inhibitory affect on the growth of Frog or DD-1
cells. Consistent with human cell line studies, resveratrol markedly inhibited both Frog and DD-1 cell growth. In both cell
lines, resveratrol induced the cleavage of caspase 3 and increased the expression of phospho-p53, two markers of apoptosis.
Resveratrol, but not metformin, increased the growth inhibitory effect of doxorubicin. These findings suggest that resveratrol may have growth inhibitory effects against canine HSA and could be a potential adjunct therapy to doxorubicin use in
canine HSA patient treatment.
Research Grant: Pre-K Career Development Award - CTSI, University of Minnesota [or] Start-up Fund,
College of Pharmacy, University of Minnesota
Student Support: College of Veterinary Medicine, University of Minnesota [or] The Skadron Family Fund
Respiratory responses to short term exposure to second hand smoke in a Guinea pig asthma model
Currie Carothers, Alexandra Noel, Zakia Perveen, Kelsey Legendre, Fabio Del Piero, Arthur Penn
Department of Comparative Biomedical Sciences (Carothers, Noel, Perveen, Penn) and Department of Pathobiological
Sciences (Legendre, Del Piero) School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA 70803
Second hand smoke (SHS) exposure is linked to increased risk of asthma, lung cancer, and chronic obstructive pulmonary
disease. Despite the current knowledge of the detrimental effects of SHS, there remains a paucity of research that elucidates
that effects that SHS has in domestic household pets. Recent surveys suggest that if pet owners were equipped with information about the deleterious effects SHS has on their pets, they would be motivated to change their smoking habits. Guinea
pigs have been proven to be excellent models for the study of asthma. For this reason, we used Guinea pigs as models for
household pets in order to determine if short-term SHS exposure, 4 hours of exposure a day for three weeks, would have
deleterious pulmonary effects on a companion animal. Our objectives were to assess respiratory immune response in Guinea
pigs that are subjected to short term SHS exposure and to evaluate changes in gene expression of Guinea pigs exposed to
SHS. After exposing 8 Guinea pigs to filtered air and 8 to an average of 10mg/m3 SHS for 21 days, the animals were sacrificed. Lung lavage and tissue collection were performed. The groups were then compared on the basis of histopathologic
changes to lung tissue, cells present in bronchoalveolar lavage fluid, and genetic changes involving the expression of IL-5
and IL-8.
118
Biomechanical risk factors in the development of osteochondrosis in Standardbred pacers and trotters
Samantha R. Carter, Christine T. Lopp, Annette M. McCoy
College of Veterinary Medicine, University of Illinois at Urbana-Champaign, Urbana, IL
Osteochondrosis (OC) is a developmental orthopedic disease in which the process of endochondral ossification is disrupted.
Although the etiology of OC is not clearly understood, there are genetic and environmental risk factors (i.e. diet, exercise)
that can increase the likelihood of developing the disease. In previous work, it was found that pacers and trotters had a predilection to develop hock OC lesions at different locations. Trotters were more likely to develop medial malleolus lesions,
whereas pacers developed distal intermediate ridge of the tibia lesions. The reason behind these differences is unknown, but
biomechanical differences between the pace and the trot may play a role. We hypothesize that the amount of time young
foals spend pacing or trotting in the field correlates with the subsequent lesions that develop, and with their severity. The
aim of this study is to quantify spontaneous activity of Standardbred pacer and trotter foals and correlate these findings with
differences in development and progression of OC lesions. Horses will be followed prospectively from 2 to 12 months of
age. Field observations will be made weekly, and hock radiographs taken every two months. Observations of activity (sleeping, nursing, walking, trotting/pacing, cantering/galloping) will be recorded at 30 second time intervals for a minimum of
2 hours per foal per week, focusing on the duration of time spent at either the pace or trot. At each radiographic time point,
4 standard views of hock radiographs will be taken and evaluated for the presence or absence of OC lesions. We expect the
lesions to be dynamic (i.e. capable of forming or healing) until 8 months of age and our work to contribute to knowledge of
OC pathophysiology.
Research Grant: Morris Animal Foundation D16EQ-311 and USDA Hatch Funds
In vitro evaluation of cell mediated immune responses to respiratory antigens in high and low risk beef calves
Kaycee R. Cash, David J. Hurley, and Brent C. Credille
Food Animal Health and Management Program, Department of Population Health, College of Veterinary Medicine,
University of Georgia, Athens, GA
Bovine respiratory disease (BRD) is the leading cause of morbidity and mortality in North American beef cattle. Stocker
cattle are at particularly high-risk of developing BRD as they have moved through multiple auction markets and often
have little health history. To prevent BRD in these cattle, vaccines are often administered upon arrival to stocker facilities.
Currently, little evidence exists to support this practice. The objective of this study was to compare cell-mediated immune
responses of low-risk, single source beef calves to high-risk, multiple source commingled calves. Complete blood counts
(CBC) were performed and peripheral blood mononuclear cells (PBMC) isolated using standard protocols. PBMCs were
stimulated in vitro with Concavalin A, BVDV-1 (strain NADL), BVDV-2 (strain A125), BHV-1, Mannheimia haemolytica,
and Pasteurella multocida. CBC parameters, lymphocyte proliferative responses, secretion of IFN-g and IL-17 into cell
culture supernatant, and expression of inflammatory cytokine genes were measured and compared between groups. Total
white blood cell and monocyte count were significantly higher in high-risk compared to low-risk cattle. Stimulation indexes
for concavalin A (1.28 vs 9.41), BVD 2 (0.25 MOI, .668 vs 1.42), M. haemolytica (.381 vs 1.58), and P. multocida (.603
vs 1.42) were significantly higher in high-risk compared to low-risk cattle. Levels of IL-17 in cell culture supernatants
were significantly higher for cells from high-risk calves stimulated with concavalin A. The findings of this study suggest
differences in immune responsiveness between low and high-risk cattle and might have implications for the development of
vaccination protocols in the future.
Student Support: Merial Ltd, Veterinary Medical Experimental Station, University of Georgia CVM
119
Effect of low starch diet on health of sloth bears (Melursus ursinus) at Cleveland Metroparks Zoo
Kaitlyn Cashin, Patricia M. Dennis, Laura Bernstein Kurtycz
The Ohio State University College of Veterinary Medicine (Cashin, Dennis), Cleveland Metroparks Zoo (Dennis,
Bernstein Kurtycz)
Captive tropical and polar bear populations are recognized as having a higher prevalence of biliary cancer than wild populations. These carcinomas are not seen commonly in domestic species. In humans, biliary carcinomas have been linked
to liver flukes and identified in populations that consume large proportions of raw fish in their diets. Chronic inflammation
may be a precursor to the formation of neoplasia and will be a focus of this study. Sloths bears in zoos are fed a diet that
is not typical of what would be consumed in the wild. The increased carbohydrates and starch in the captive diet may be
leading to increased inflammation and stereotypic behaviors. Serum inflammatory markers, insulin resistance parameters,
and behavior of the two sloth bears (Melursus ursinus) housed at the Cleveland Metroparks Zoo will be compared before
and after a lower starch diet is introduced. Specific markers to be analyzed include insulin, insulin: glucose ratio, and serum
amyloid A. Behavior observations are conducted using video recording while the bears are on and off exhibit. Continuous
sampling with focal follow is performed for coding the behavior. It is hypothesized that providing sloth bears in captivity
with a diet that is lower in starch will lead to decreased inflammation and stereotypic behaviors.
Research Grant: Cleveland Metroparks Zoo
Student Support: The Ohio State University Monahan research fellowship
Daniele T. Casper, Mingqun Lin, and Yasuko Rikihisa
Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, Ohio
Ehrlichia chaffeensisis a zoonotic bacterium that causes human monocytic ehrlichiosis when transmitted by the bite of infected ticks. Previous discoveries in the Rikihisa Laboratory have shown that E. chaffeensisbinds and enters human blood
cells using a specific bacterial surface protein, termed Entry triggering protein of Ehrlichia (EtpE), by directly binding to the
mammalian cell surface protein, DNase X. Since DNase X is known to cleave DNA, we hypothesized that DNase X activity
can be inhibited by EtpE-binding. Therefore, the purpose of this study was to examine whether EtpE can enhance gene delivery efficiency into mammalian cells. Mammalian RF/6A cells were incubated with recombinant C-terminal fragments of
EtpE (rEtpE-C), and a recombinant Ehrlichia-secreted protein Etf-1 as a negative control. Cells were then transfected with a
plasmid encoding green fluorescent protein (GFP). The amount of plasmid delivered into mammalian cells was determined
by quantitative PCR (qPCR). Results showed that the plasmid amount was slightly increased in RF/6A cells incubated with
rEtpE-C normalized with Gapdh gene. This suggests that EtpE might block the catalytic site of DNase X, allowing the
stabilization of exogenous DNA; therefore enhancing its delivery and expression into mammalian cells. These findings can
possibly improve the current method of gene therapy, which is often limited by DNA degradation.
Research Grant: Research Grant: OSU Accelerator Awards Project #29-100006
Student Support: Student Support: National Institutes of Health T35OD10977
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Natalie J Castell, Thaddeus C Deiss, and Michael F Criscitiello
Department of Veterinary Pathobiology, College of Veterinary Medicine & Biomedical Sciences, Texas A&M University,
College Station, TX.
Jawed cartilaginous fish represent the most primitive extant species which possess adaptive immune systems homologous to
those of mammals. Because of this, they act as a unique resource for investigating the evolution and function of the immune
systems found in both humans and other vertebrates. The primary objective of this project is to gain a better understanding
of an unusual and only recently discovered population of lymphocytes in the immune repertoire of the nurse shark. Previous work from this lab has found that a subset of cells within the thymus undergo functional somatic recombination which
employs immunoglobulin, specifically IgM and IgW, gene segments. These cells deviate from the traditionally distinct
lineages of T and B cells by producing and employing Ig/TCR chimeric receptors. To better understand this subpopulation,
characterization of their maturation and expression patterns is being performed using a histological approach. To visualize
and investigate the cells of interest present in the shark thymus, fluorescence in-situ hybridization has been performed to
probe for selected mRNA products. Specific mRNA probes for RAG, AID, TCRg, TCRd, TCRa, and IgW allow for classification of developing T cells as they mature and move outward through the thymus. The normalization of images and
quantification of mRNA product will be performed using ImageJ software. Neither the location and development, nor the
specific expression profiles of these B-like T cells has been demonstrated before; these different pieces of information will
collectively begin to reveal more about development pathways in the shark thymus, and give novel insights into the limits
of vertebrate B and T cell antigen recognition.
Research Grant: NSF award IOS1257829 to MFC
Student Support: Coll. Veterinary Medicine & Biomedical. Sci., Texas A&M
The proline utilization system of Brucella abortus is required for virulent infection
Mitchell T. Caudill, James A. Budnick, Lauren M. Sheehan, Christian R. Lehman, and Clayton C. Caswell
Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine, Virginia Tech,
Blacksburg, Virginia
The proline utilization system (Put) has been linked to virulence in a number of enteric pathogens. The importance of the
Put system in the intracellular pathogen Brucella abortus, however, has not been explored. In this poster, we show the Put
system, consisting of the genes putA (bab1_0518) and putR (bab1_0517), are critical for the survival of B. abortus in mice,
but not in macrophages. The Put system is composed of the enzyme proline dehydrogenase (PutA) and its transcriptional
regulator (PutR). A lacZ reporter fused to the putA promoter showed that PutR acts as an activator in Brucella, and electrophoretic mobility shift assays confirmed that PutR binds directly to the intergenic region of putA and putR. Additional
experiments are determining the exact sequence of the intergenic region bound by PutR and possible reasons for attenuation
in mice. Altogether, these data are the first to reveal that the Put system plays a significant role in the ability of Brucella
abortus to replicate and survive within its host.
Research Grant: VA-MD College of Veterinary Medicine
Student Support: American Veterinary Medical Foundation
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Development and validation of a PCR platform for the sensitive and specific detection of Bartonella rochalimae
Dennis Chan, Pedro Paulo V. P. Diniz
Western University of Health Sciences CVM, Pomona, CA
We developed a sensitive and specific PCR platform for detecting the recently discovered Bartonella rochalimae - a pathogen capable of causing bacteremia and other life threatening symptoms in humans and animals. Currently its detection relies
on the amplification of DNA using genus-level primers. This method is ineffective if other species of Bartonella are present
in the same sample, as seen frequently in vectors. We manually designed three new primers, targeting the genes Intergenic
Transcribed Spacer (ITS), citrate synthase (gltA) and b subunit of bacterial RNA polymerase (rpoB), which has a limit of
detection of 5 Genomic Equivalent(GE)/reaction, 5 GE/ reaction and 10 GE/reaction, respectively. These three PCR assays
were designed to generate amplicons of 425bp to 864bp in size, which is then sequenced for species confirmation. At optimal PCR conditions, our platform did not amplify DNA from 10 other distinct Bartonella spp., as well as human, canine
and feline genomic DNA. Despite highly stringent PCR conditions, the ITS and gltA PCR assays generated an amplification when tested against a very high concentration of B. clarridgeiae (1x106 GE/reaction). The distinction between these
species, however, can be done using restriction enzymes that only cleave B. rochalimae DNA, such as BspHI, CciI, PagI or
RcaI. Conversely, the rpoB PCR assay was highly specific and did not amplify the concentrated B. clarridgeiae samples.
Therefore, the combined use of these new assays can serve as a tool for the specific detection of B. rochalimae in humans,
animals and vectors. This will not only improve our diagnostic capabilities, but also enhance our knowledge in the ecological and epidemiological studies of such pathogens.
Fluoroscopic determination of thoracic dimensional changes in healthy dogs
Jennifer C. Chan, Lynelle R. Johnson, Craig S. Brown, Rachel E. Pollard
University of California Davis School of Veterinary Medicine, Davis, CA
Lung compliance, defined as the change in lung volume for a given change in airway pressure, is typically measured under
anesthesia using sophisticated pulmonary mechanics analysis. Interstitial lung diseases of various etiologies reduce lung
compliance. Fluoroscopy, a noninvasive imaging technique, allows real time assessment of lung area changes. We hypothesized that percent change in thoracic area between inspiration and expiration would be repeatable in healthy dogs and a proxy
for pulmonary compliance. 44 dogs free of respiratory disease were positioned in sternal recumbency. Fluoroscopic imaging
of the thoracic cavity was performed for 10 respiratory cycles during tidal respiration. Maximal inspiratory and expiratory
images were selected for 3 respiratory cycles. Percent change in lung area was calculated in 2 ways. “Total” measurement
yielded the number of pixels within the entire thoracic cavity. “Segmented” measurement excluded the mediastinum and
cardiac silhouette. Percent change in pixels between inspiration and expiration was calculated and averaged for each dog.
To compute a reference range, parametric (normal and log normal) and non-parametric methods (percentile, bootstrapping
statistics) were evaluated, and use of percentiles was chosen as the preferred method. The “total” measurement yielded a
median of 12.5% with a 95% confidence interval of 8.91-23.97%, and the “segmented” measurement had a median of 20.8%
with a confidence interval of 14.32-37.61%. This study confirmed the utility of fluoroscopy as a noninvasive method for
assessing lung area changes. Further studies are needed to assess the utility of this method in dogs with respiratory disease.
Student Support: Students Training in Advanced Research (STAR) program at the University of California, Davis
122
Characterizing the mechanisms of action of novel chemical inhibitors of Mycobacterium smegmatis growth
A’Jah A. Chandler, Emily E. Juzwiak, Robert J. Fillinger, Robert B. Abramovitch
Department of Microbiology and Molecular Genetics, Michigan State University, East Lansing, MI
Mycobacterium tuberculosis is the bacterium that causes the infectious disease, tuberculosis. One-third of the world’s population is infected with this bacterium. The key feature of tuberculosis disease is the granuloma, which is caused when
alveolar macrophages, infected with bacteria, are surrounded by other cells to form a protective barrier in the lung. The
granuloma provides a hypoxic and acidic environment that restricts bacterial growth and spread. These two factors cause the
bacterium to go into a dormant phase, in which antibiotics do not inhibit them as well. In attempt to further understand the
mechanism of action of drugs, our lab is trying to characterize chemical compounds that may inhibit the growth of dormant
bacterium. We hypothesize that in a hypoxic environment, the chemical compounds PH003, PH017, and tyrphostin will
inhibit the growth of this bacterium. The effects of chemical compounds were tested on a nonpathogenic strain, dormant
M. smegmatis. The dormant M. smegmatis was generated simply by adding a small concentration of the bacteria to a flask,
along with growth medium, and sealing the flask so there is no access to oxygen. Bacterial growth consumes the oxygen and
causes a gradual onset of hypoxia and bacterial dormancy. Hypoxia did in fact slow the growth of M. smegmatis. However,
the chemical compounds, PH003, PH017, and tyrphostin did not inhibit growth of the bacterium under these conditions.
Characterizing these chemical compounds may lead to new drugs that can possibly shorten the time it takes to treat tuberculosis. However, M. smegmatis may not be the best model for this study, so this experiment will be tested against M.
tuberculosis as well.
Research Grant: NIH-NIAID 1-R01-AI11660J-01
Fecal toxin cytotoxicity as a measure of clinical disease in mice infected with Clostridium difficile R20291
Kevin Chang, Jenessa A. Winston, Stephanie A. Montgomery, Casey M. Theriot
Department of Population Health and Pathobiology, College of Veterinary Medicine, North Carolina State University,
Raleigh, NC (Chang, Winston, Theriot) Department of Pathology and Laboratory Medicine, Lineberger Comprehensive
Cancer Center, University of North Carolina School of Medicine, Chapel Hill, NC (Montgomery)
Clostridium difficile is a Gram-positive, spore-forming anaerobic bacterium and is the major cause of antibiotic-associated
diarrhea and colitis in humans. C. difficile infection (CDI) is a leading nosocomial infection and poses an urgent threat to
public health. Clinical disease is mediated by C. difficile toxins, TcdA and TcdB, and virulence varies by strain. Previous
mouse studies have demonstrated a correlation between toxin induced cytotoxicity and histopathologic changes within the
cecum, however it is unclear whether a similar correlation exists between these parameters and fecal cytotoxicity. Our hypothesis is that fecal cytotoxicity correlates with both cecal cytotoxicity and histopathologic lesions in mice infected with C.
difficile R20291, a clinically relevant strain. C57BL/6J mice (5-8 wk) were treated with cefoperazone and later challenged
with 105 C. difficile spores. Mice were monitored for clinical signs of disease, including weight loss, for the first 4 days. At
the time of necropsy (Day 0, 2, and 4), samples were collected to measure toxin cytotoxicity in cecal contents and feces,
using a Vero cell cytotoxicity assay, and histopathologic changes to the cecum. Using spearman correlation, we observed a
correlation between fecal cytotoxicity and both cecal cytotoxicity (r=0.9999) and cecal inflammation (r=0.9997). Based on
these findings, fecal cytotoxicity levels may be an accurate alternative measure of other disease parameters in the gut during
CDI. Utilizing fecal cytotoxicity as a surrogate measurement for cecal cytotoxicity and histopathology will eliminate the
need for multiple necropsies throughout a study, thus reducing the total number of animals required for future experiments.
123
Comparison of NIBP measurements obtained in a veterinary clinic versus a home setting in hypertensive dogs
Jenna L Charles, Anderson F da Cunha, and Mark J Acierno
Department of Clinical Sciences, Louisiana State University SVM, Baton Rouge, LA
Hypertension is an important systemic illness of dogs that can result in kidney, eye, heart and brain damage. Typically, veterinarians rely on non-invasive blood pressure measurements to diagnosis hypertension as well as monitor patient response
to therapy. Previous studies have demonstrated that healthy dogs may experience a blood pressure elevation in response to
stressful situations such as veterinary visits. We hypothesized that some patients diagnosed as hypertensive may actually
be normotensive while others may be hypertensive but with exaggerated blood pressure measurements while they are at
the veterinary clinic. This could lead to unnecessary treatment of some patients and overmedication of others.This study
will determine if blood pressure taken in a clinical situation is significantly different than pressure measured in the home
environment in dogs diagnosed with hypertension. Furthermore, we will explore if these differences result in unnecessary
medication. Findings to be presented.
Student Support: Louisiana State University SVM Dean’s Fund
Systematic literature review and elicitation of expert opinion to assess the 1997 FMD epidemic in Taiwan
Aolei Chen, Amy C. Kinsley, Andres M. Perez
Department of Veterinary Population Medicine, College of Veterinary Medicine, University of Minnesota, St. Paul, MN
Foot and mouth disease (FMD) is a highly contagious disease affecting cloven-hooved animals that causes devastating economic losses to affected countries. Because the United States (US) has been FMD-free since 1929, little is known regarding
the likely size and extension of an FMD epidemic in the country. Disease models may help predicting epidemic dynamics
and identifying appropriate control measures, and utilizing field data from FMD epidemics are essential to increase model
accuracy. Here, a combination of systemic review of scientific literature and field interviews was used to study the features
of the 1997 FMD epidemic in Taiwan. Formal sources of information included 1 book, and 6 peer reviewed-publications
with 3 in English and the other 3 in Chinese. Informal sources of information included the interviews, in Taiwan, of 5 individuals with field experience at the time of the epidemic. Formal and informal sources of information agreed in many
aspects except some key epidemic parameters, such as the date of first introduction of FMD virus into Taiwan, percentage
of under-reported cases, and effectiveness of control measures. Because formal and informal sources are subject to different types of bias, a combination of literature review and elicitation of expert opinion may help to reconstruct the history of
epidemics more accurately than if a unique source was used. Results here will help the parameterization of an FMD spread
model in U.S. swine farms with the ultimate goal of supporting early detection and mitigation of impact of a hypothetical
FMD epidemic in the U.S.
Research Grant: National Pork Board and the MnDrive program
Student Support: Minnesota Veterinary Medical Foundation and College of Veterinary Medicine, University of MN
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In vitro endothelial cell wound healing in response to red wine polyphenols resveratrol and quercetin
Colin T Chu, Merilyn H Jennings, and Tammy R Dugas
Department of Comparative Biomedical Sciences, Louisiana State University School of Veterinary Medicine,
Baton Rouge, Louisiana
Treatment of coronary artery disease (CAD) faces frequent complications, as the bare metal stents (BMS) used to reopen
atherosclerotic arteries often cause chronic restenosis, or vessel renarrowing. Stent placement initially damages the surrounding endothelium and delays endothelial cell (EC) healing, leading to a chronic and invasive inflammatory reaction
that causes long term vascular smooth muscle cell (VSMC) proliferation and vessel restenosis. Drug-eluting stents (DES)
coated with antimitogenic agents reduce restenosis, but also inhibit EC proliferation, further delaying wound healing and
leading to late in-stent thrombosis. Two red wine polyphenols, resveratrol (R) and quercetin (Q), are of interest as potential
DES agents. R has been shown to promote EC wound healing, while R & Q have been shown to interact more than additively to reduce injury-induced neointimal hyperplasia in a mouse model, both in vivo. We hypothesized that additionally R
& Q would interact to enhance EC wound healing in vitro. Artificial wounds were induced to cell-seeded microscope slides
and initial wound width was measured. Cells were then exposed to R, Q, and a 1:1 combination of R+Q for 18h. Wound
width was then reevaluated and % wound healing calculated for each treatment. Under static culture conditions we found
that R, Q, and R+Q negatively impact EC wound healing. However, ongoing studies are aimed at assessing R and Q effects
on wound healing under laminar flow conditions that mimic blood flow.
Effect of FABP1/SCP-2/SCP-X Gene Ablation (TKO) on the Endocannabinoid System in Mice
Sarah Chung, Gregory G. Martin, Lawrence J. Dangott, Huan Huang, Danilo Landrock, Kirsten Landrock,
Avery McIntosh, Sherrelle Milligan, Friedhelm Schroeder, Ann B. Kier
Department of Physiology and Pharmacology (Chung, Huan, Martin, McIntosh, Schroeder) and Department of Pathobiology, Texas A&M University, College Station, Texas, USA (Chung, Landrock, Milligan, Kier) Department of Biochemistry
and Biophysics, Texas A&M University, College Station, Texas, USA (Dangott)
The endocannabinoid (EC) pathway influences neurophysiology (behavior, appetite, pain, and inflammation), embryonic
development, and hepatic lipogenesis. Abnormal EC regulation is associated with pathophysiology such as obesity, non-alcoholic fatty liver disease (NAFLD), and alcoholic fatty liver disease (AFLD). Much of current EC research has focused on
pain and inflammatory components of the EC pathway, particularly in novel pain therapies. Cannabinoid receptors (CB) in
the EC pathway and enzymes that degrade EC ligands such as anandamide (AEA) and 2-arachidonoylglycerol (2-AG) are
the focus of new therapeutics. While this has produced several drugs in clinical trials, many have severe side effects. Our
research focuses on an alternative strategy-targeting cytosolic EC binding/chaperone proteins. Our studies have shown that
liver fatty acid binding protein (FABP1) and sterol carrier protein-2 (SCP2) are major EC binding proteins in both the brain
and liver. Moreover, FABP1 ablation (LKO) indirectly increases brain EC by removing FABP1’s ability to facilitate hepatic
clearance of ARA (precursor of AEA and 2-AG). However, unaltered levels of SCP2 complicate interpretation of this phenotype. The objective of our study was to show how ablation of FABP1 and SCP-2/SCP-X genes (triple knock-out, TKO)
affects EC levels in mice. TKO mitigated the LKO-induced increase in AEA and 2-AG in brain as well as the LKO-induced
increase of 2-AG (but not AEA) in liver. TKO also ameliorated some of the effects of high-fat diet on EC levels. These findings indicate the importance of both FABP1 and SCP-2 as regulators in the EC system, making them potential therapeutic
targets.
Research Grant: NIH RO1 DK41402 (Schroeder, Kier)
Student Support: AVMA/AVMF 2nd Opportunity Summer Research Scholarship
125
Characterization and optimization of ex vivo T cell expansion for re-directed T cell therapy
Megan Clark, M. Kazim Panjwani, Nicola Mason
Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA
Non-Hodgkin’s lymphoma (NHL) is the most common hematological malignancy and second most common cancer in
dogs. While combinations of chemotherapy and systemic cytotoxic agents remain the treatments of choice for canine NHL,
such therapy is often ineffective, as 85-90% of patients relapse with drug-resistant disease within 6 to 9 months of diagnosis
and treatment. Adoptive immunotherapy using re-directed T cells to recognize and kill tumor cells represents a powerful
strategy to target primary, metastatic, and relapsing neoplastic disease. In this approach, autologous T cells are activated,
transduced to express chimeric antigen receptors (CAR) specific for a tumor cell surface antigen, and expanded ex vivo prior
to adoptive transfer back into the patient. We have developed CD20-targeting CAR T cells to use in dogs with relapsed B
cell lymphoma and our early results are encouraging. However, the success of this approach depends upon the ability of
these cells to engraft and persist for the lifetime of the host. The T cell subsets used for adoptive transfer have been shown to
have a significant impact on engraftment and expansion of CAR T cells in vivo, with T cells of central memory phenotype
resulting in superior engraftment. Using flow cytometry and qPCR, we are evaluating which cytokine conditions are most
favorable for the ex vivo generation of canine memory T cell subsets. This identification of which T cell growth condition
promotes optimal engraftment and CAR T cell persistence will be essential for long term, effective therapies for dogs with
relapsed or refractory B cell lymphoma.
Research Grant: The Richard Lichter Charity for Dogs
Student Support: Supported by NIH grant T35 OD-010919 and a grant from Merial
Early effects of ivermectin on intraperitoneal Brugia pahangi infection in Meriones unguiculatus
William A. Clark, Adrian Wolstenholme, Chris C. Evans, Katie M. Day, Michael T. Dzimianski, Kaori Sakamoto,
and Andrew R. Moorhead.
Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA
Lymphatic filariasis is a parasitic disease, caused by Wuchereria bancrofti and Brugia malayi, which affects over 120 million
people worldwide. Recent evidence suggests that ivermectin, an anthelmintic used in global filariasis elimination programs,
may prevent nematodes from secreting proteins that permit evasion of the host immune response. Mongolian gerbils, or
jirds (Meriones unguiculatus), are an accepted rodent model for filariasis research. We hypothesized that exposure of B.
pahangi third-stage larvae (L3) to ivermectin would reduce larval yields and motility after intraperitoneal (IP) infection in
jirds. Male jirds were infected IP with 100 B. pahangi L3s; 12 jirds received L3s that were cultured overnight with ivermectin in propylene glycol, while the remaining 12 jirds received L3s that were cultured with vehicle only. Within each group,
half of the jirds received an IP ivermectin injection upon infection, while the remaining jirds were treated with vehicle. At
13-17 days post-infection, jirds were euthanized and their peritoneal cavities were lavaged to collect surviving fourth-stage
larvae (L4). Recovered L4s were counted, and larval motility was measured using the “Worminator” video imaging system.
Data were analyzed using pairwise Mann-Whitney U tests with Bonferroni correction. Mean larval recovery and motility
were not significantly different among the four treatment groups (control L3/control jird: 12.8 larvae; control L3/treatment
jird: 14.8 larvae; treatment L3/control jird: 10.2 larvae; treatment L3/treatment jird: 9 larvae). These findings suggest that
parasite clearance from the jird peritoneum due to ivermectin treatment does not occur in the first two weeks of infection.
Research Grant: NIAID R01AI103140
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Alterations in myofilament-associated signaling complexes in dogs with naturally occurring DCM.
Samantha B Clarke, Ilka Lorenzen-Schmidt, Yuanhua Cheng, Michael Regnir, M Lynne O’Sullivan, W Glen Pyle
Centre for Cardiovascular Investigations, Department of Biomedical Sciences (Clarke, Lorenzen-Schmidt, Pyle) and the
Department of Clinical Science (O’Sullivan), Ontario Veterinary College, University of Guelph, Guelph, ON Department
of Bioengineering, University of Washington, Seattle, Washington (Cheng, Regnir)
Dilated cardiomyopathy (DCM) is the most common heart disease in large and giant breed dogs. Studies of canine DCM
have failed to definitively identify the cause. Cardiac myofilament mutations are the most common cause of familial DCM
in humans and myofilament dysfunction contributes to disease progression. No studies have examined cardiac myofilament
alterations in naturally occurring canine DCM. We hypothesized that cardiac myofilament function is impaired along with
altered levels of myofilament-associated intracellular signaling molecules in spontaneous canine DCM. Dogs were assessed
by echocardiography and myocardial samples used to isolate myofilaments. Myofilament function was assessed using an actomyosin Mg2+-ATPase assay and contraction/relaxation measured using myofibrils. Myofilament proteins were separated
by gel electrophoresis and stained with ProQ Diamond for total phosphorylation or immunoblotting for protein levels. DCM
myofilaments had increased ATP use and impaired contraction/relaxation compared to non-failing samples. Desmin and tropomyosin phosphorylation were increased in DCM as were total desmin protein levels. Total troponin I phosphorylation was
unchanged, but S23/S24 phosphorylation was reduced in DCM. Myofilament-associated PKCd and PKC-zeta were elevated
in DCM, PKCe was reduced, and PKCa changed. Myofilament associated PP1g, PP2A-A and PP2A-C were elevated, while
PP1a was reduced in DCM hearts compared to controls. These data are the first to show cardiac myofilament modifications
in naturally occurring canine DCM, and support our hypothesis that alterations in cardiac myofilaments contribute to DCM.
Research Grant: OVC Pet Trust
Anatomy of the lower respiratory system of the African Grey Parrot (Psittacus erithacus erithacus)
Ashley O. Cleveland, Alexandra K. Ford, Emma R. Schachner
School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA (Cleveland) School of Veterinary Medicine,
Louisiana State University, Baton Rouge, LA (Ford) Department of Cell Biology & Anatomy, School of Medicine,
Louisiana State University Health, New Orleans, LA. (Schachner)
The avian respiratory system has been the focus of detailed study since Thomas Henry Huxley’s mid 19th century descriptions of the lungs of the brown kiwi and the duck. Since then, the gross and microscopic anatomy of the lower respiratory
system of numerous species of birds has been described in detail based upon dissections, casts, and light and scanning
electron microscopy. Recent advances in digital imaging techniques, specifically computed tomography (CT) and magnetic
resonance imaging (MRI) have revolutionized the study of anatomy, and powerful visual analysis programs have facilitated
the creation of three-dimensional (3D) digital models of skeletal and soft tissue structures in vertebrates for use in basic biology and paleontology. Surprisingly, these methods have not been fully integrated into veterinary medicine, despite the clear
benefits for diagnostic and surgical planning purposes. Here we propose to map out the 3D anatomy of the lower respiratory
system in a popular avian pet, the African Grey Parrot (Psittacus erithacus erithacus), with the aim of investigating the relationships between the thoracic organs in situ, the extent of normal variation in these structures, and to develop an atlas of
the pulmonary structures relative to topographical and radiological skeletal landmarks. The results of this project will be integrated into the Grey Parrot Anatomy Project being conducted by avian clinicians and researchers at the University of Utah.
Student Support: 2016 Merial/NIH Veterinary Summer Scholars Program
127
Effects of IL-10 modulation on nerve pathology in C. jejuni induced Guillain-Barre mouse models
Matthew M. Cluett, Jean Brudvig, Linda Mansfield
Comparative Enteric Diseases Laboratory, Departments of Large Animal Clinical Sciences and Microbiology and
Molecular Genetics, College of Veterinary Medicine, Michigan State University.
Guillain-Barre syndrome (GBS) is an auto-immune disorder of the peripheral nervous system that results in ascending
flaccid paralysis. 7% of patients die and 20% have long term disability, yet no effective therapeutics exist. Antecedent infections with several pathogens, most prominently Campylobacter jejuni, have been linked to development of autoreactive
antibodies thought to mediate GBS through cross-reaction with peripheral nerve gangliosides. Mouse models that develop
GBS nerve lesions subsequent to C. jejuni infection are needed to elucidate the pathogenesis and to establish targets for new
treatments. We hypothesized that interleukin (IL)-10 deficient mice orally inoculated with a C. jejuni GBS patient strain
will have higher numbers of macrophage infiltrates in peripheral nerves, and higher autoantibody levels will correlate with
more severe nerve lesions. BALB/c wild-type (WT) and IL-10-/- mice were inoculated with a GBS-associated C. jejuni strain
260.94 or sham inoculated (vehicle)(4 groups, 10 mice/group) and sacrificed after five weeks. Sciatic nerves and lumbar
dorsal root ganglia were dissected from representative subsets of each of the four treatment groups, embedded en bloc, sectioned and labeled immunohistochemically with an F4/80 macrophage marker. Macrophage infiltration in DRG was scored
quantitatively using morphometry. Infected WT and IL-10-/- mice had significant increases in anti-C. jejuni antibodies, yet
there was no correlation between autoantibody levels and the number of infiltrating macrophages. Infected IL-10-/- mice had
a trend to significantly higher macrophage infiltrates, but there was no significant difference between sham and infected WT
mice. Results from remaining mice are pending.
Research Grant: These studies were funded by NIAID, NIH, Department of Health and Human Services, under Grant No.
U19AI090872 Enterics Research Investigational Network, Cooperative Research Center.
Student Support: NIH Grant No T35OD016477 to Michigan State University
Effect of temperature on Veronaea botryosa growth and cytotoxicity to Acipenser transmontanus skin cell lines
Denver Coleman, Susan Yun, and Esteban Soto
Department of Medicine and Epidemiology, School of Veterinary Medicine, University of California, Davis, Davis, CA
95616, USA.
Sturgeon aquaculture is particularly important since wild populations are significantly affected due to overfishing, habitat
destruction and pollution. In the Pacific North-west, white sturgeon (Acipenser transmontanus) culture is a multi-million
dollar industry. The production of globally recognized high quality caviar made this product one of the few aquaculture-generated exports for the United States. However, the emergent fungal pathogen, Veronaea botryosa, causes high mortality rates
that result in significant economic losses for the industry. This melanized mold is the etiologic agent of systemic phaeohyphomycosis, also known as “fluid belly”. Cultured white sturgeons are subjected to a range of temperatures during their
lifetime, especially in aquaculture where higher temperatures accelerate growth and decrease time to reproductive maturity
and caviar production. Little is known about the environmental conditions that allow V. botryosa to colonize and infect
fish. Infection is prevalent in adult white sturgeon, particularly when the water temperature is above 188C. Environmental
conditions such as water temperature can play critical roles in the development of both the innate and adaptive immunity.
A better understanding of the immune response of cultured white sturgeon at different life stages, temperatures, and water
quality would provide direct benefits for the aquaculture industry. In this research, we investigated the germination and in
vitro growth of V. botryosa at 20 and 258C in seven different nutrient media. Additionally, we present a novel in vitro culture
method to investigate the interactions between V. botryosa spores with a previously reported white sturgeon skin cell lines
at 15, 20 and 258C.
Research Grant: University of California Davis
Student Support: Student Teaching and Research, UC Davis, CA
128
Effect of mesenchymal stem cell conditioned media on TGF-b1 exposed equine fibroblasts
Jillian A. Condrey, Merrilee Thoresen, and John F. Peroni
Department of Large Animal Medicine, College of Veterinary Medicine, University of Georgia, Athens, GA 30602
Exuberant granulation tissue (EGT) is a complication in distal limb wounds of horses leading to delayed healing and poor
outcomes. Normal equine skin contains ~ 1 ng/mL of transforming growth factor-b1 (TGF-b1) which increases to ~ 6 ng/
mL following injury. In distal limb wounds of horses TGF-b1 remains elevated 2-4 weeks following injury and likely contributes to the formation of EGT. Mesenchymal stem cells (MSCs) have been shown to contribute to wound healing and
cutaneous homeostasis due to their inherent paracrine anti-inflammatory effects. Our aim was to determine if treatment with
bone marrow derived MSC conditioned media (CM) or activated conditioned media (ACM; MSCs pretreated with 10 ng/
mL tissue necrosis factor a (TNFa)) would counteract the pro-fibrotic effect of high levels of TGF-b1 over a 96-hour culture
period. Equine dermal fibroblasts from 2 horses were cultured in fibroblast conditioned media (control), CM, or ACM with
either 1 or 6 ng/mL of TGF-b1. Cells were harvested at 24, 48, and 72 hours following treatment and assayed for viability,
proliferation, and gene expression. Our preliminary results showed that fibroblasts treated with CM and ACM decreased
their expression of pro-fibrotic genes, a-smooth muscle actin and type I collagen, as compared to fibroblasts cultured in control media. We also found that the total number of cells at each time point and the doubling times remained consistent across
all treatments, while the results of our viability assay were variable and did not exhibit any consistent trends. In conclusion,
our results demonstrate that MSCs could potentially ameliorate the stimulatory effect of TGF-b1 which contributes to the
sustained fibroproliferative response typical of EGT.
Research Grant: Regenerative Biological Therapies Laboratory, College of Veterinary Medicine, University of Georgia
Emily Courville, Olalekan M. Ogundele, Charles C. Lee
LSU School of Veterinary Medicine, Baton Rouge, La
Mutations in the CNTNAP and 16p11.2 genes in humans results in a form of Autism Spectrum Disorder (ASD), characterized by abnormal language development, social interactions, and repetitive behavior/restricted interests. Transgenic CNTNAP2 and 16p11.2 knockout mice exhibit ‘similar’ ASD behavioral phenotypes. In addition, brain development is altered
in these knock-out mice. Specifically, the normal formation of the cerebral cortex is altered, such that excessive ectopic neurons are found in the lower cortical layers. Neurons in lower cortical layers 5 and 6 of the auditory cortex typically project
to the medial geniculate body of the thalamus. However, it is possible for the neurons to project to other areas of the cortex.
We hypothesize that the extra ectopic neurons will also project to the thalamus, although these cells could also project to
different areas of the cortex. This excessive ‘drive’ of the thalamus could result in some of the restrictive and repetitive behaviors observed in these animals.
Research Grant: NIH/NIMH grant R03MH104851, Louisiana Board of Regents grant RCS RD-A-09
Student Support: Kenneth F. Burns Foundation
129
Role of inhibitor of differentiation (Id) proteins in fibrosis of the human cornea
Amanda Cox, Govindaraj Anumanthan, Sudhanshu P. Raikwar, Suneel Gupta, Ratnakar Tripathi, Prashant R. Sinha,
and Rajiv R. Mohan
One-Health One-Med Ophth Research Ctr, Vet Med and Surgery and Biomed Sciences, Coll of Vet Med(Cox,
Anumanthan, Raikwar, Gupta, Tripathi, Sinha, Mohan), and Mason Eye Inst., School of Medicine (Mohan),
Univ of Missouri, Columbia, MO, Harry S. Truman Memorial Veterans’ Hosp, Columbia, MO (Anumanthan,
Raikwar, Gupta, Tripathi, Sinha, Mohan)
Ocular trauma and infection result in corneal fibrosis and vision loss in human and veterinary patients. Fibrosis is induced
by Transforming Growth Factor b1 (TGFb1) as a part of a complex healing process and is characterized by the differentiation of fibroblasts to myofibroblasts. Inhibitor of differentiation (Id) genes are known to regulate cellular differentiation.
Recently, we have characterized expression of Id genes 1-4 in the human cornea and human corneal fibroblasts (HCF), and
found that TGFb1 modulates Id protein expression in HCF. However, the role of Id genes in HCF differentiation and corneal
healing is still unknown. The purpose of the present study is to understand how Id proteins regulate corneal fibrosis in the
presence of TGFb1. We hypothesize that Id2 and Id3 over-expression in HCF functions as a molecular switch and drives
cellular fate for corneal fibroblast transdifferentiation. Primary cultures of HCF were established from donor human corneas
and delivered Id2 or Id3 gene either via Lipofectamine 3000 or jetPEI nanoparticles. Id gene overexpressing HCF cultures
were identified. These clones were grown in the presence/absence of TGFb1, and analysis of fibrotic markers aSMA,
Collagen I, IV, and fibronectin by qPCR and immunocytochemistry is underway. We expect to detect decreased expression
of fibrotic markers and differentiation in cultures with overexpression of Id2 and Id3 compared to positive controls (HCF
exposed to TGFb1). This information will increase our understanding of the interaction of TGFb1 with Id2 and Id3 genes
in the cornea, may offer a novel gene-based modality for treating blindness, and benefiting patients.
Research Grant: 1I01BX00035701 (RRM) Veteran Health Affairs, Washington, DC, RO1EY17294-07 (RRM) National
Eye Institute, Bethesda, MD, and Ruth M. Kraeuchi Missouri Endowed Chair Ophthalmology Fund (RRM)
Student Support: Zoetis Animal Health
TAM Receptor Expression Is Altered During Zika Virus Infection in Pregnant Mice
Nathan Crilly, Meghan Vermillion, Jun Lei, Yahya Shabi, Victoria Baxter, Michael McLane, Diane Griffin,
Andrew Pekosz, Irina Burd, Sabra Klein
W. Harry Feinstone Department of Molecular Microbiology and Immunology (Crilly, Vermillion, Baxter, Griffin,
Pekosz, Klein), Bloomberg School of Public Health; and Department of Gynecology and Obstetrics, School of Medicine
(Lei, Shabi, McLane, Burd); Johns Hopkins University, Baltimore, MD.
Zika virus (ZIKV) infection in pregnant women is associated with placental infection and congenital disease, and represents
a major global public health threat. Research using both in vitro and in vivo models shows that the placenta serves as a site
of productive ZIKV replication. However, cell entry receptors in the placenta remain ill defined. TAM receptors are tyrosine
kinase receptors that can be used as cell entry receptors for flaviviruses, including ZIKV. We explored the in vivo role of
TAM receptors in the context of ZIKV infection using a mouse model, hypothesizing that if TAM receptors are used for
cell entry in the placenta, then TAM receptor expression would be upregulated in placenta compared with other peripheral
tissues, and TAM receptor expression would be altered in the placenta during ZIKV infection. Timed-pregnant CD1 mice
received an intrauterine inoculation of either 106 TCID50 ZIKV or vehicle on embryonic day (E)10. Two days after infection,
spleen and placenta were collected and tested for ZIKV RNA copies and infectious titers, as well as the relative expression
of TAM receptors and associated proteins (Gas6, Axl and Mertk). In both ZIKV-infected and mock-infected tissues, the
expression of the TAM receptor Axl was upregulated in the placenta compared with the spleen. Moreover, Axl expression
was greater in the placentas of infected dams compared to mock-infected dams. Together, these results suggest that Axl may
play a role in the preferential targeting of ZIKV infection to the placenta during pregnancy and therefore may represent a
promising therapeutic target.
130
The genetics of familial hypertrophic cardiomyopathy in the Siberian cat
Kathryn F. Crombie, Nicholas Gustafson, Leslie A. Lyons
College of Veterinary Medicine, University of Missouri - Columbia, Columbia, MO (Crombie, Gustafson, Lyons),
School of Veterinary Medicine, University of California - Davis, Davis, CA (Lyons)
Hypertrophic cardiomyopathy (HCM) is the most common inherited cardiovascular disease. To date, a large number of
genetic studies have established that HCM is caused by mutations in 11 or more genes encoding the contractile components
of the sarcomere or adjacent Z-disc. Despite the identification of numerous causative mutations, the pathogenic and genetic
processes are still poorly understood, making a large animal model of familial HCM especially useful. Causative mutations
have been identified in Maine Coon and Ragdoll cats, however, HCM is thought to be inherited in other breeds, including
the Siberian cat, where the novel causative element remains unknown. Utilizing the candidate gene approach to focus on
associations between genetic variation within multiple important sarcomeric and cardiac genes, this study aims to identify
the causative element of HCM within the Siberian cat breed. Genes implicated in 8 cardiac diseases, including HCM, were
selected based on their known biological, physiological, and/or functional relevance to cardiomyopathies. Non-synonymous
single nucleotide polymorphisms were identified in genes encoding Ankyrin 2 (ANK2), Syntrophin Alpha 1 (SNTA1), and
Titin (TTN) in an affected Siberian cat, all absent from 9 unaffected genomes and 73 of unknown HCM status. Variants
within these genes will be compared between affected cats, cats with an affected family member, and cats diagnosed as
normal upon ultrasound. By identifying the causative element in Siberian HCM, the development of a genetic test for the
disease could help eradicate the disease in future Siberian populations and provide an additional large animal biomedical
model of HCM.
Assessing mental health and identifying stressors among LSU School of Veterinary Medicine students and alumni
Carolyn Croughan, Stephanie Johnson, Michael Kearney, Carmen Asuaga, and Joseph Taboada
Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Louisiana State University, Baton Rouge,
Louisiana
Previous research found that the suicide rate among veterinarians may be as high as four times the general population and
twice that of other healthcare professions. The American Veterinary Medical Association recently found that 67% of vet
students nationally have experienced depression. Survey Monkey was used to survey 246 Louisiana State University School
of Veterinary Medicine (LSU SVM) students and 495 LSU SVM alumni. The two surveys included questions on substance
abuse and major stressors, as well as the Center for Epidemiologic Studies Depression Scale (CES-D) and the Mental Health
Inventory (MHI) Anxiety Subscale. The CES-D revealed that 45% of students’ scores indicated potential symptoms of clinical depression and the MHI Anxiety Subscale showed that 70% of students reported symptoms of anxiety. Finances were
identified as the highest overall stressor among LSU SVM students and alumni. Besides finances, alumni report having the
most difficulty with balancing work with personal and family life and dealing with patients with unexpected outcomes. In
2014, the National Association of State Public Health Veterinarians and the CDC found that one in six veterinarians have
considered suicide. This is three times the national adult mean. LSU SVM alumni fell slightly below the national average.
Alumni also reported that suicide is a problem in the veterinary profession.
Research Grant: LSU School of Veterinary Medicine Summer Scholars Program National Institute of Health
Student Support: LSU School of Veterinary Medicine Summer Scholars Program National Institute of Health
131
Cancer Stem Cell-Targeted Vaccine Induces Antibodies Against Conserved Vaccine Antigens
Ross E Cutcliffe, Jade Kurihara, John Coy, Dan Regan, Steven Dow, and Amanda Guth
Center for Immune and Regenerative Medicine, Department of Clinical Sciences, and Flint Animal Cancer Center,
Veterinary Teaching Hospital, Colorado State University, Fort Collins CO
Cancer stem cells (CSCs) are increasingly becoming recognized as a critical cellular subcomponent of tumors. They are
believed to be involved in cancer initiation and cancer therapy resistance, due to their self-renewing capabilities and various
means of averting chemotherapy and radiation therapy. Due to their inherent resilience, there has been difficulty in developing a therapeutic approach directed toward the destruction of CSCs. However, our lab developed a novel CSC vaccine,
consisting of tumor cell lysates derived from tumor cells grown in CSC-enriched conditions, that significantly slowed tumor
growth in mice compared to a conventional vaccine, and that is now undergoing a canine clinical trial. We hypothesize that
dogs vaccinated with our CSC vaccine will induce a potent antibody response against novel, immunogenic CSC antigens. In
order to substantiate this hypothesis, we analyzed vaccines created from CSC-enriched stem cells compared to conventional
tumor lysate vaccines in murine models. Western blotting was used to identify immunogenic protein bands, combined with
microarray analysis to determine genes of interest. This led to the discovery of conserved immunogenic antigens that were
upregulated in the CSC-vaccinated mice in comparison to conventional-vaccinated mice, and substantial differential gene
expression across both groups. Serum samples from the canine CSC-vaccine clinical trial were collected for Western blot
analysis, resulting in the detection of novel protein bands. Further analysis of these proteins could lead to increased understanding of CSCs, and the development of a more potent option of immunotherapy to efficiently initiate an immune response
specified toward the destruction of CSCs.
Research Grant: Shipley Foundation Program in Applied Regenerative Medicine and the National Institute of Health T35
The prophalactic effect of acetylsalicylic acid on astrogliosis in a mouse model of scrapie
Jo Daney, Damani Bryant, Caitlin Feiock, Davis Seelig
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, Saint Paul, MN
The misfolded form of the prion protein (PrPD) causes fatal neurodegenerative disease. Prion diseases have been identified in many species including humans (Creutzfeldt-Jakob disease), cows (BSE), and sheep (scrapie). Prion diseases are
characterized by deposition of PrPD throughout the central nervous system, neuronal and synaptic loss, spongiosis, clinical
dementia, and neuroinflammation represented by astrogliosis and microgliosis. There is no treatment for prion disease and
survival after diagnosis in humans is less than 2 years. Acetylsalicylic acid (ASA) has many anti-inflammatory effects including COX2 inhibition, NF-kB regulation, and inhibition of ERK-12 pathways. These pathways contribute to astrogliosis
and increased expression of glial fibrillary protein (GFAP). We tested whether or not the administration of ASA would impact astrocytosis in a scrapie model system. Mice were intraperitoneally infected with 20 ml of 10% scrapie-positive brain
homogenate and oral ASA was administered either as a prophylactic (i.e. prior to prion infection) or as an intervention (i.e.
after the mice began to show clinical signs based upon rotarod testing). All mice were euthanized at terminal disease and
their brains were harvested, fixed, sectioned, and stained for GFAP using fluorescent immunohistochemistry. GFAP expression was quantified using a custom macro in Image J. To date, the treated and untreated mice show statistically significant
increased GFAP expression compared to controls (two-way ANOVA, p-value#.05), but a significant effect of ASA has yet
to been seen. Future study will include IHC for PrPD deposition and microgliosis, an increase in sample size (from n=3 to
n=6), and analysis of time-matched sacrifice group.
Student Support: National Institutes Of Health, Award Number T35OD011118
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Barbara L. Daniels, John P. Buchweitz, John B. Kaneene, Heather N. DeFore, Vilma Yuzbasiyan-Gurkan,
and Daniel K. Langlois
Department of Small Animal Clinical Sciences (Daniels, DeFore, Yuzbasiyan-Gurkan, Langlois), Department
of Pathobiology and Diagnostic Investigation (Buchweitz), and the Center for Comparative Epidemiology (Kaneene),
College of Veterinary Medicine, Michigan State University, East Lansing, MI
Exposure to lead as a major public health problem has garnered considerable attention due to the water crisis in Flint, MI
following a water source change in 2014. For over one year, many residents and their pets were exposed to lead-contaminated water, which could result in a multitude of health problems. Although most relief efforts focused on human health, pets
were also at great risk of lead exposure, exemplifying the need for a “One Health” approach. The purpose of our study was
to investigate blood lead levels (BLLs) and clinical abnormalities in dogs residing in Flint, MI. We hypothesized that dogs
in Flint would have higher blood lead levels (BLLs) than those of a non-Flint control population. We further hypothesized
that BLLs would be higher in young dogs and in those with clinical signs consistent with lead toxicity. Blood samples were
obtained via public screening clinics for Flint, MI dogs and via a veterinary clinic in East Lansing, MI for control dogs. Lead
concentrations were measured using inductively coupled plasma mass spectrometry. Overall, Flint dogs had significantly
higher BLLs than control dogs, and younger dogs had higher BLLs than older dogs. Spatial analysis suggested some associations between BLL’s and city ward water lead levels. Associations between clinical signs and BLLs are likely, but data
analysis is ongoing. These results suggest that pets residing in Flint were impacted by the water crisis and further magnifies
the importance of considering both human and animal well-being in a public health crisis.
Research Grant: Michigan State University Gifted and Endowed Research Funds
Student Support: NIH Grant T35OD016477 to Michigan State University
Elizabeth Daugherty, R. W. Stich, Kelly Straka, Shuping Zhang
University of Missouri College of Veterinary Medicine, Department of Veterinary Pathobiology
Missouri represents an epicenter of tick-borne pathogens. Among the contiguous United States, Missouri is a national leader
in reported cases of Rocky Mountain spotted fever and ehrlichiosis. Tick-borne disease also represents a highly underdiagnosed condition in human and veterinary medicine (Gleim et al., Fihn et al.). This phenomenon makes epidemiological
surveillance of limited use. Improved veterinary surveillance of wild mammal populations, food animal operations, and
companion animals could serve as a foundation for a more robust tick-borne pathogen monitoring system. Our goal is to
develop a polymerase chain reaction (PCR)-based diagnostic test panel for detecting a diverse range of tick-borne pathogens in mammalian blood as well as in tick tissues. The objective of this project is to develop and to utilize a single positive
control for PCR assays to detect tick-borne Anaplasma, Borrelia, Ehrlichia and Rickettsia species. Our approach to this
work was to select previously validated primer sets for incorporation in tandem into a synthetic gene block. PCR parameters were optimized for each primer set prior to adaptation to real-time PCR. Primer analytical sensitivity with the g-block
control will also be determined. Pathogens within field-collected ticks must be inactivated for safe handling in the lab, so
this g-block control will also be tested in the context of different tick fixation and template preparation methods. Lastly,
ticks opportunistically collected from white-tailed deer in Missouri will be screened with this panel of PCR assays. Future
objectives will include using these procedures to adapt and perfect this tick-borne pathogen assay for detection of additional
disease agents in Missouri wildlife.
Research Grant: Missouri Department of Conservation, MU Veterinary Medical Diagnostic Laboratory, USDA National
Institute of Food and Agriculture, Animal Health project 1008645
Student Support: CVM Department of Veterinary Pathobiology
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Purification and stability screens of conserved virulence factor A in Streptococcus pyogenes
Jose De La Espriella, Suryatej Akavaram, Katherine Turnbull, Kyu Hong Cho, Gabriela C. Perez-Alvarado
and Brian M. Lee
Biomedical Sciences, (De La Espriella, Akavaram, Turnbull, Lee) and Veterinary Microbiology and Preventive Medicine
(Perez-Alvarado), College of Veterinary Medicine, Iowa State University, Ames, IA 50011. Department of Biology,
College of Arts and Sciences, Indiana State University, Terre Haute, IN 47809 (Cho).
Conserved virulence factor A (CvfA), also known as RNase Y, is a transmembrane protein in Streptococcus pyogenes and
is conserved in many Firmicutes bacteria such as Bacillus subtilis and Staphylococcus aureus. S. pyogenes causes mild superficial skin infections, pharyngitis and necrotizing fasciitis. CvfA has been shown to influence the expression of virulence
factors in S. pyogenes by regulating mRNA processing leading to either degradation or activation for translation. CvfA has
four domains which include a transmembrane domain that allows CvfA to attach to the cell membrane, a coiled-coil region
that is used for dimerization, a KH domain that is used to recognize nucleic acids, and finally an HD domain that may cleave
the nucleic acids. The goal is to characterize the structure and function of the KH and HD domains of CvfA. Two constructs
were designed, one containing the KH domain (residues 157-320) and another with the KH and HD domains (residues
157-515). To prepare for structural studies, the CvfA constructs were purified using affinity, ion exchange, and gel filtration
chromatography. Purified CvfA constructs were then tested under various solution conditions using thermofluor assays to
measure protein stability. The most stable solution conditions for the smaller KH domain construct (CvfA 157-320) will
be used for structural and functional studies by NMR spectroscopy. The thermofluor assays for the dual KH-HD domain
construct (CvfA 157-515) have provided optimal buffer, pH, salt and additives for crystallization screens that may allow us
to collect X-ray diffraction data and determine the structure of both the KH and HD domains.
Research Grant: NIH-NIGMS (7 R15 GM101603-02 MPI Lee, Pérez-Alvarado and Cho)
Student Support: Center for Advanced Host Defense Immunobiotics and Translational Comparative Medicine (Lee).
Do male sticklebacks use visual or olfactory cues to assess the history of a potential mate?
Marion Dellinger, Jennifer Hellmann, Alison M. Bell
Department of Animal Biology, School of Integrative Biology, University of Illinois at Urbana-Champaign, Urbana, IL
Differential allocation occurs when individuals alter their reproductive investment based on their mate’s traits. Previous
studies have shown that male threespined sticklebacks (Gasterosteus aculeatus) alter their behavior toward a potential mate
depending on her previous experience with predators: when a male encountered a female who had been exposed to predators
while yolking her eggs, he showed fewer courtship displays as well as less care to her offspring relative to an encounter
with an unexposed female. Although both chemical and visual cues mediate conspecific interactions and mate choice in this
species, we do not know the mechanism males use to detect previous predator-exposure in mates. To test the extent to which
those cues aid in male detection of female predation exposure, we compared male courtship behavior across four treatments
in a 2x2 repeated measures design manipulating a potential mate’s previous experience (predator-exposed or not) as well as
the presence or absence of olfactory cues from predator-exposed females. We confined females to a transparent box providing visual cues only, while olfactory cues were added via water from the tanks of predator-exposed females (experimental)
or predator-naive ones (control). We predicted that if males use olfactory cues to assess female predation exposure, males
exposed to experimental water would behave differently compared to males exposed to control water, regardless of female
predation status. If males use visual cues, however, only the female status should alter male courtship. Determining the relative importance of visual versus chemical cues will help us better understand the mechanisms by which males adjust their
behavior based on female phenotypes.
Research Grant: NSF grant #1121980, NIH grant #GM082937
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Does in utero exposure to domoic acid, found in seafood, cause epilepsy?
Fanny Demars, and Paul Buckmaster
VetAgro-Sup School of Veterinary Medicine, Lyon, France (Demars) and Department of Comparative Medicine, School
of Medicine, Stanford University, Stanford, CA (Demars, Buckmaster)
Harmful blooms of domoic acid-producing algae are a growing problem worldwide. As a glutamate receptor agonist, domoic acid can cause gastrointestinal and neurological symptoms. Among other species, humans are at risk of severe consequences as the Canadian incident in 1987 of massive ‘amnesic shellfish poisoning’ showed. After eating contaminated
mussels, 3 patients died, and ~40% of patients developed neurological symptoms like amnesia and seizures. Despite the
establishment of tests for seafood, concerns persist as monitoring doesn’t occur everywhere. Plus, consequences of chronic
exposure to levels below the regulatory limit are unknown. Partially based on previous rodent studies done by others, it has
been proposed that dietary exposure to doses of domoic acid that are subclinical in pregnant women may damage the fetal
hippocampus and initiate epileptogenesis. To test that hypothesis, we are using continuous, telemetric video-EEG seizure
monitoring for 1-month-long periods in mice exposed to 1.2 mg/kg of domoic acid at embryonic day 13, which coincides
with the peak of hippocampal neurogenesis. Preliminary data reveal epilepsy in 4/14 mice exposed in utero to domoic acid
and in 0/4 vehicle-treated mice. In epileptic mice, 9-32 seizures/month were observed, and average seizure duration was
31-56 seconds. Nissl and Timm staining are being performed to test for pathological changes typical of temporal lobe epilepsy such as hippocampal neuron loss and mossy fiber sprouting. This study will help evaluate the potential risk of in utero
domoic acid exposure in humans.
Research Grant: NSF and NIH (NIEHS)
Student Support: School of Medicine, Stanford University
Joseph Devereaux, K. Kazmierczak, G. Breur
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, IN
Gait maturation is the developmental process by which young animals develop the pattern of walking that is considered
normal in adults. This is an area that has been explored in human medicine but much less is known about gait maturation in
animals. This information is valuable to determine if a deviation from what is considered normal in healthy adults is pathologic or simply a product of development. In human medicine this information has allowed for early intervention and better
outcome in diseases that cause gait abnormalities in children due to early detection and treatment. The goal of this study is
to identify the differences, if any, between the gait of healthy mature dogs and that of puppies and to determine when gait
has fully matured. We hypothesized that gait will be mature before the age at which we begin testing. Kinetic and kinematic
data was gathered on five puppies starting as young as possible continuing until 32 weeks of age. Trends were examined
in the data set and compared to reference intervals (RIs) previously established on adult dogs. Most parameters examined
fell within the reference ranges for healthy adults. The most obvious and well supported trend seen in the data is a negative
association between the coefficient of variation (CV) of many measured variables and the puppies’ age. At 20 weeks of age
all parameters appear to have normalized. A similar trend has been seen in children up to 3-4 years of age and has been
attributed to continued neural development. Given these preliminary findings it seems justified to conduct further research
into canine gait development and possibly establish reference ranges for healthy puppies to better identify when pathology
is present in young animals.
Student Support: Purdue University Summer Research Fellowship
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Evaluating the effect of Fortiflora probiotic therapy on feline Tritrichomonas foetus infection
Rachel E. Dickson, David A.Bemis, M. Katherine Tolbert
Department of Small Animal Clinical Sciences (Dickson, Tolbert) and Department of Biomedical and Diagnostic Sciences
(Bemis), College of Veterinary Medicine, University of Tennessee, Knoxville, TN
Tritrichomonas foetus (Tf) is a highly prevalent enteric protozoa, which causes chronic diarrhea in domestic cats. Ronidazole is the only effective treatment for feline Tf infection. However, drug toxicity and resistant strains of Tf warrant identification of alternative therapies. Probiotics have been purported to be effective against Tf infection, however little to no
work has been done to prove their efficacy. Probiotics may aide in controlling pathogens by influencing the intestinal environment, antagonizing pathogens, and modulating the host’s immune system. Demonstrating that probiotics reduce clinical
signs and recurrence of Tf infection would completely change the approach to treatment of feline Tf. Therefore, the study
objectives were to evaluate the effect of probiotic therapy on Tf growth, as well as adhesion and cytotoxicity of Tf to the
intestinal epithelium using an in vitro co-culture model. The commercially available veterinary probiotic containing Enterococcus faecium (Ef), was used as it is a widely-available and is single species. The lyophilized product was reconstituted,
inoculated on blood agar plates, and verified to be Ef by several methods. Co-culture with Ef inhibited Tf replication at all
concentrations tested with ratios as low as 10 Tf:1Ef resulting in complete inhibition of Tf growth. This effect is thought
to be the result of a pH change induced by Ef or its secretions. Indeed, when the pH was neutralized, no significant effects
of Ef or its secretions were observed on TF growth or adhesion to intestinal epithelial monolayers in the co-culture model.
Further work is ongoing to determine if the Ef-containing probiotic would be an effective adjunctive for treatment of feline
trichomonosis.
Student Support: UTCVM Center of Excellence
Evaluating the role of female sex hormones on left ventricular mechanics in aortic-banded mini-swine
Madeleine Dionne, Jessica A. Hiemstra, T. Dylan Olver, Jenna C. Edwards, Tracy Swanson, Pamela K. Thorne,
Jan R. Ivey, Craig A. Emter
University of Missouri- Columbia, College of Veterinary Medicine, Department of Biomedical Sciences
Heart failure with preserved ejection fraction (HFpEF) comprises approximately 50% of all human heart failure cases. HFpEF has twice the incidence in woman compared to men, despite previous studies suggesting estrogen may be protective
against the development of heart failure. The purpose of this study was to determine the role of female sex hormones on
left ventricular (LV) mechanics of the heart following chronic pressure-overload using two-dimensional speckle-tracking
echocardiography (2D-STE). Recent studies have proposed increasing the clinical use of 2D-STE given its practicality and
reduced expense compared to MRI. For this study, 28 Yucatan miniature swine were placed into four groups (N=7/group):
intact control (CON), intact aortic-banded (AB), ovariectomized control (CON-OVX), and ovariectomized aortic- banded
(AB-OVX). Six segments of the LV and septum were generated from apical four-chamber and short-axis two-dimensional
views (acquired at the mitral-valve and apex levels) and averaged to determine global strain, strain rate, and displacement in
the longitudinal, transverse, radial, and circumferential dimensions over three cardiac cycles. Torsion was calculated as the
difference between mitral and apical end systolic rotation (degrees) and normalized to both LV hypertrophy (wall thickness)
and end diastolic chamber length. We hypothesized that global strain would be reduced primarily in the longitudinal direction in the AB group, and that this finding would be further impaired by the loss of female sex hormones.
Research Grant: NIH/NHLBI R01 HL112998 (CAE)
Student Support: NIH/NHLBI R01 HL112998 (CAE)
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Optimizing antimicrobial use for mastitis to combat antimicrobial resistance on US dairy farms
Ann E. DiPastina, Karun Kaniyamattam, and Yrjo T. Grohn
Michigan State University College of Veterinary Medicine (DiPastina), East Lansing, MI; Cornell University College
of Veterinary Medicine (Kaniyamattam and Grohn), Ithaca, NY
The development of resistant microbes is a threat to animal and human health which can be abated, but not reversed. Antimicrobial resistance (AMR) has been propagated by the inconsiderate application of antimicrobials in humans, animals, and
plants. As such, combating AMR requires a concerted effort from animal and plant agricultural industries as well as human
and veterinary medical professionals. System dynamics is a useful method of describing complex issues such as AMR and
has been used to study complex diseases such as diabetes and HIV. In this project, we used system dynamics to describe
the relationship between antimicrobial use (AMU) and AMR. Although AMR is a global concern, we focused on AMU
within the United States, particularly in animal agriculture, and with an emphasis on the dairy industry. Our first objective
was to create a concept map which identifies factors contributing to AMU at the farm level as well as the consequences of
AMU. From this map, we designed a dynamic model to simulate the relationship between AMU for mastitis (an assumed
risk factor for AMR) and farm profitability. Ideally, optimal AMU on dairy farms improves profit and minimizes AMR in
both animals and humans. Understanding AMU in animal agriculture is a necessary step toward developing effective regulations to control their use. With this approach, we have gained a very specific understanding of AMU at the farm level on a
typical US dairy farm. This model can be extended beyond the farm to investigate the effect of AMU on the onset of AMR
in consumers of dairy products. This project is one highly focused component of the unified effort to slow the development
of resistant microbes.
Microneutralization assay: an economical method for determining vaccine recommendations for swine influenza
Janna Draper, Samuel Yingst, Katrina Miller
College of Veterinary Medicine, Purdue University, West Lafayette, IN (Draper) Assistant Professor of Molecular
Diagnostics, Animal Disease Diagnostic Laboratory, Purdue University, West Lafayette, IN (Yingst) Laboratory
Supervisor of Virology, Animal Disease Diagnostic Laboratory, Purdue University, West Lafayette, IN (Miller)
Following diagnosis of Influenza A, rapid antigenic characterization of virus isolates and/or antibody is vitally important
in order to make informed decisions regarding control measures. The current characterization techniques include hemagglutinin inhibition (HI) and virus neutralization (VN). These tests yield good results but HI is not as sensitive or specific, and VN is substantially slower than the microneutralization assay. The microneutralization assay is used to detect
virus-specific antibodies of serum to influenza virus or to antigenically characterize influenza viruses using a reference
panel. The aim of this research, was to adapt the microneutralization assay so that it can be used to identify which vaccine
is the best to prevent the spread of Influenza A. The microneutralization test consists of three stages; (1) virus titration,
(2) virus microneutralization, and (3) ELISA. Variables that had to be taken into account and modified included media,
viruses, incubation times, Madin-Darby Canine Kidney (MDCK) prep, and ELISA plates. To establish the test in the lab,
control serum was tested against H3N2 and H1N1. The results have been varied and further testing is required. Additionally, the control serum will be tested against H3N2 and H1N1 using HI. The results of the MN and HI will be compared
and analyzed. The results of our testing so far indicate that the microneutralization assay is achievable in the lab setting
and can be utilized in the laboratory for determining which vaccine is best suited to prevent an Influenza outbreak.
137
Stephie-Anne Cassagnol Duliepre, Catalina Pereira, Ellen Sunmin Hong, and Robert Weiss
Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY
Due to the volume of endogenous and exogenous stresses encountered by our cells every day, a process known as the DNA
damage response (DDR) plays a critical role to help maintain proper genomic integrity. The DDR has been shown to allow
germ cells to successfully advance through meiosis, particularly Prophase I, where double-stranded breaks (DSBs) occur
to facilitate pairing of homologous chromosomes and crossing over. A central part of the DDR pathway is the RAD9ARAD1-HUS1 (9-1-1) complex, a heterotrimeric clamp that acts as a molecular scaffold for DNA repair and checkpoint signaling activation, through interaction with several DNA damage response proteins. It has been previously established that
testis-specific inactivation of Hus1 or Rad9a leads to; germ cell depletion, and severe meiotic defects. Recently paralogs of
9-1-1 subunits, RAD9A and HUS1, termed RAD9B and HUS1B, have been proposed to form alternative, non-canonical
meiotic 9-1-1 complexes (RAD9B-RAD1-HUS1B and RAD9B-RAD1-HUS1), which are still poorly understood. We used
Hus1b knockout mice (Hus1b-/-), in conjunction with Hus1 inactivation (hypomorphic Hus1 allele or conditional knockout
targeted to the testis) to better understand the roles of these alternative 9-1-1 complexes. We hypothesize that disruption
of multiple 9-1-1 complexes with loss of both Hus1b and Hus1 will disrupt proper progression through meiotic Prophase
I. Supporting our hypothesis, preliminary results show germ cell loss in both males (increased lumen space) and females
(decreased primordial follicle counts). These studies will help elucidate the diverse roles of the 9-1-1 complex on fertility
and overall DNA stability.
Student Support: Office of Graduate Education – Cornell University College of Veterinary Medicine
Elizabeth M. Dunfield, Neil I. Christensen, and Michelle M. Turek
Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI
Radiotherapy has played a significant role in providing curative or palliative treatment for both veterinary and human oncology patients. Conventional radiotherapy involves daily low-dose treatments over the course of several weeks. A novel
method of radiation delivery, Stereotactic Radiotherapy (SRT), utilizes specialized technologies to improve accuracy and
precision so that tumors can be effectively treated in one to three high-dose fractions, whilst avoiding normal surrounding
tissues. The potential benefits of SRT include equal or improved tumor control, decreased cumulative radiation doses, and
improved sparing of normal tissue. In human medicine, a task group recently assessed SRT practices and used available
literature to report “best practice guidelines.” Similar guidelines in the veterinary field are not available, and a major impediment to their development is a lack of knowledge regarding current practices. To better understand SRT usage, we conducted a survey of veterinary radiation oncologists in the United States, Canada, Europe, and Australia. We hypothesized
that there is significant variation among veterinary radiation facilities providing SRT in terms of equipment types, treatment
planning and protocols, treatment goals, and quality assurance methods. Response accrual is ongoing, but we also expect to
determine the prevalence of SRT use in veterinary medicine and to evaluate future directions of SRT in clinical and research
settings. With the gained understanding of how, when and why SRT is utilized in veterinary medicine, we hope to provide a
foundation upon which best practice guidelines can be established to further advance this novel method of cancer treatment.
Student Support: University of Wisconsin School of Veterinary Medicine
138
Determination of causes of mortality in captive psittacines submitted to the Ontario Veterinary College
T. Eagalle, N. Nemeth, H. Beaufrere, L. Susta
University of Guelph, Department of Health Sciences (Beaufrere) and Department of Pathobiology (Eagalle, Nemeth,
Susta), Ontario Veterinary College, Guelph, Ontario, Canada
Psittacines are increasingly common in households, aviaries and zoological collections. With the advancement of avian
medicine, necropsy and biopsy are becoming increasingly important for accurate disease diagnosis. However, comprehensive information about disease conditions in birds, including pathological descriptions, is often limited. We retrospectively
reviewed diagnostic data from psittacines submitted to the Ontario Veterinary College (OVC) to assess baseline prevalence
and pathological features of the most commonly diagnosed diseases. Diagnostic data from psittacine necropsy and biopsy
cases submitted to the OVC and Animal Health Laboratory from 1995-2015 were reviewed. Data included signalment and
morphologic diagnoses, which were categorized and ranked to identify primary cause(s) of death. Eighty-three species
(n=1149) were represented, with the African grey and cockatiel as the most common. Infectious agents were most commonly identified as the primary cause of death (n=409; 35.6%), including 216 viral (52.8%; some suspect), 132 bacterial
(32.3%), and 61 fungal (14.9%) infections. The most common respective etiologies were bornavirus (n=154), Mycobacterium spp. (n=18), and Aspergillus spp. (n=18). Neoplasia was diagnosed in 100 cases (8.7%), which most commonly involved
the alimentary tract (n=18); squamous cell carcinoma was the most frequently diagnosed neoplasia (n=9). Better recognition
and understanding of diseases that impact psittacines will aid clinicians and pathologists to formulate more effective differential diagnoses and targeted diagnostic procedures, ultimately improving the health of captive psittacines in Ontario and
elsewhere.
Research Grant: Pet Trust Fund
Student Support: Andrea Leger Scholarship
The Effect of Sandstorm Particles on Antigen-Presenting Dendritic Cells
Danielle Ebelle, Nicholas Buglak, Bin Wang, Furong Deng, Ning Li
Department of Pathobiology and Diagnostic Investigation, Michigan State University, East Lansing, MI Institute
of Reproductive and Child Health/ministry of Health Key Laboratory of Reproductive Health, School of Public Health,
Peking University, Beijing, China
Asian Sand Dust (ASD), also known as sandstorm, is a severe dust wind originating primarily in the Gobi desert in China.
This has led to serious health concerns among many Asian countries due to the worsening of air pollution in China; i.e.
extremely high levels of particles with aerodynamic diameter < 2.5 mm (PM2.5) with high polycyclic aromatic hydrocarbon
(PAH) content. Exposure to PM2.5 is a risk factor for cardiopulmonary diseases including asthma. We hypothesize that
ASD-associated PM2.5 can induce oxidative stress [activation of heme oxygenase-1(HO-1)] and inflammatory responses
[increase in interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNFa) release] in dendritic cells (DC), one of the central
regulators of the immune response. PM2.5 samples were collected in 2004 in Baotou, China during non-sandstorm weather
days and during two sandstorms. DC were stimulated with these particle samples for 16 hours. Our results show that all samples increased IL-6 release and induced HO-1 expression, while all but one sample increased the amount of TNFa released.
However, the strongest effects were observed in the cells exposed to one of the non-sandstorm weather day samples. The
metal contents of our samples positively correlated with the observed inflammatory response: silica, nickel, and copper with
both cytokines; cadmium with TNFa; and chromium with IL-6. Transition metals can undergo redox reactions leading to
cellular oxidative stress. No positive correlation was found between the PAH content and the inflammatory response. Taken
together, this suggests that the particle-associated metals may be the major component of our ASD samples responsible for
the inflammatory response of DC leading to asthma exacerbation.
Research Grant: International Visiting Research Scholar Award and Startup Fund
139
Breast cancer cell growth mediated by EGFR/HER1 expression is regulated by p53 suppressor function
Allison L. Ehrlich and Bimal K. Ray
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia, MO
Our goal is to prevent cell growth in breast cancer by suppression of EGFR gene expression. By preventing cell growth, we
hope to stop the spread and improve treatability of breast cancer. How p53 affects EGFR transcription remains unknown.
We hypothesize that large amounts of p53 protein available for binding silences EGFR transcription. We cultured and prepared extracts of one normal human mammary epithelial cell line, MCF10A, and three breast cancer epithelial cell lines,
MCF7, MDA-MB-231, and MDA-MB-468. Gel electrophoresis and western blot were used to measure p53 levels in each
cell line, followed by an electrophoretic mobility shift assay to look for protein binding of p53 to EGFR promoter. EGFR
expression is high in breast cancer cells. A 40 kDa derivative of p53 is present at a significantly lower level in breast cancer
cells compared to normal breast epithelial cells. Normal cells had high levels of binding of p53 derivative, while significantly lower levels of such binding were found in breast cancer cells, particularly in MDA-MB-231 and MDA-MB-468 which
both overexpress EGFR. These results indicate that loss of p53 binding to EGFR promoter is at least partly responsible for
EGFR overexpression in breast cancer cells. Increased binding of p53 or its derivative may reduce EGFR expression. This
suggests that EGFR could be targeted directly for breast cancer treatment in the future.
Research Grant: This work was supported in part by the United States Army Medical Research and Material Command
Grant W81XWH-09-1-0084 and the University of Missouri Faculty Research Activity Grant
Student Support: Student was supported by the University of Missouri Faculty Research Activity Grant
Role of Neural Inflammatory Processes in Estradiol Enhancement of Cocaine Self Administration in Female Rats
Nicole Emmitt, Luis Martinez, Robert Meisel, Paul Mermelstein
College of Veterinary Medicine (Emmitt), Department of Neuroscience (Martinez) Department of Neuroscience
and Graduate Program of Neuroscience (Meisel and Mermelstein), University of Minnesota, Minneapolis, MN
Drug abuse is an enduring problem across all populations, but women initiate drug use at younger ages and progress to
clinical addiction faster than their male counterparts. Estradiol is thought to be a major contributor to this sex differences,
conveying increased vulnerability to addiction by remodeling neuronal synapses in brain reward pathways. Although the
precise mechanisms remain unclear, microglia have been identified as potential mediators of this drug-induced plasticity.
In response to chronic exposure to cocaine, M1 glial cells are activated leading to release of the pro-inflammatory factor
TNF-a. Release of TNF-a leads to internalization of AMPA receptors and a dampening of behavioral responses to cocaine.
In females, estradiol acts to convert M1 pro-inflammatory glial cells to M2 anti-inflammatory glial cells, thus resulting in
decreased TNF-a levels and a subsequent decrease in neural inflammation. We hypothesize that the switch from M1 to M2
microglia is critical to the enhanced addictive behaviors seen in females. To test this hypothesis, ovariectomized female
rats received jugular vein catheters and sc estradiol capsules, and then were allowed to freely administer cocaine for 6 hrs
each day for 10 days. Control animals received no catheter implants (and did not self-administer cocaine) and/or no hormone capsule implants. At the conclusion of testing, brain tissue was collected and processed to identify activation state of
microglia (via immunohistochemistry for Iba1) as well as expression of TNF-a. These studies will provide insight into the
role of neuroinflammatory processes in addictive behaviors in females, and help identify therapeutic targets for treating this
disease in women.
Research Grant: National Institutes of Health, Award Number R01 DA035008
Student Support: National Institutes of Health, Award Number T35 OD011118
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Make food great again: effect of protein source and amount on whole-body metabolism and weight gain
Danielle Engel, Kristy Du, and Justin Rhodes
College of Veterinary Medicine (Engel), Department of Animal Sciences (Du) and Department of Psychology (Rhodes),
University of Illinois at Urbana-Champaign, Urbana, IL
Recent studies have shown that over 66% of the U.S. adult population is overweight or obese. Approximately 30% of dogs
and cats are overweight, indicating that obesity is a significant problem in the pet population as well. Protein is an important
element of the daily diet. Previous studies indicated that certain types of protein, such as egg white (EW) and wheat gluten
(WG), have satiating and anti-satiating effects on rats, respectively. The goal of this study was to determine if animal and
plant-based protein sources affect metabolism, leading to weight loss or slower weight gain in rats over a 30-day period.
Higher metabolic rates and slower weight gain were expected in WG treatment groups due to a predicted increase in energy
expenditure based on amino acid composition. After an overnight fast, adult Sprague-Dawley rats were provided 30 minutes
of access to a meal of manipulated protein source and amount. These meals were either 20% or 35% WG or EW and the
amount was calculated to be 10% of daily food consumption. Food intake and body weight were recorded daily. EchoMRI
was used to measure body composition on days 1, 15, and 30. Columbus Lab Animal Monitoring System (CLAMS) cages
were used to analyze respiratory exchange ratio (RER) and heat produced during the first and last 3 days of the study. No
significant difference was found between treatment groups with regard to weight gain and metabolism. RER data suggested
that as rats adapted to their new diet regimen, protein became a larger source of fuel. Although the results did not support
the overall hypothesis, RER data suggest that long-term effects of high protein diets on protein digestion merit further study.
Research Grant: National Institute of Diabetes and Digestive and Kidney Diseases, NIH, DK082609
Student Support: Office of the Director, NIH, T35 OD011145
Recreating geophagy in a captive psittacine species, Myiopsitta monachus
Gena Esposito, EV Voltura, Donald J Brightsmith
Texas A&M University College of Veterinary Medicine & Biomedical Sciences, College Station, TX
Geophagy, the consumption of soil, is a phenomenon that occurs on every continent except Antarctica and amongst many
different taxa. In birds, the most common reasons cited for geophagy behavior include protection from dietary toxins and
mineral supplementation. In Southeastern Peru, 28 bird species as well as many insects and mammals have been documented consuming soil. Geophagy has substantial costs in terms of time, energy, and predation risk. Preferred soils in the region
are high in sodium and toxin-binding clays, but these variables are tightly correlated making isolation difficult. Average
food plants are low in sodium, but little data is available on sodium requirements for parrot species. The objective of this
study was to establish a model system of geophagy in a captive psittacine species, Myiopsitta monachus (Quaker Parakeet),
and to better understand what might drive parrots to consume clay. Quaker parrots (n=20) were housed in pairs and fed
either a control diet containing 1500 ppm sodium or a low sodium diet containing 300 ppm sodium. All birds were offered
a kaolin-based clay containing 2500 ppm sodium similar to preferred clays consumed in SE Peru. Daily food and clay food
intake was monitored for each cage. Birds were monitored daily for adverse health events and received physical exams before, during, and after the study. This experimental geophagy model will be used to better understand geophagy in parrots in
SE Peru and establish daily sodium requirements for Quaker parrots. The findings of this and future studies will help identify
keystone resources for Peruvian fauna and will provide valuable information to enhance the health of captive psittacines.
Research Grant: none
141
Glutathione S-transferase-mu polymorphisms and cancer risk in boxers
Rachel M. Falsey, Joanne L. Ekena, Katherine R. Luethcke, Lauren A. Trepanier
Department of Medical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI
Glutathione S-transferases (GSTs) are enzymes found in the liver and other organs that detoxify reactive environmental carcinogens and endogenous free radicals. In humans, low-activity polymorphisms in the genes encoding GST-mu, GST-theta,
and GST-pi are risk factors for lymphoma. Boxers are one dog breed with a higher incidence of lymphoma, but the underlying risk factors are not understood. Our laboratory is interested in the role of GST polymorphisms in breed-specific
cancers in dogs, and how these interact with environmental exposure. For this study, we are focusing on GST-mu (encoded
by GSTM1 in boxers. We hypothesize that deleterious GSTM1 polymorphisms predicted to decrease enzyme expression
or activity will be over-represented in the boxer breed as a whole. To test this, we are resequencing all 8 GSTM1 exons in
genomic DNA from pure-bred boxers and non-boxer dogs, and analyzing variants for predicted functional effects using in
silico software. To date, we have begun screening for GSTM1 variants in 28 boxers and have banked DNA from an additional 45-50 non-boxer dogs. Identification of GSTM1 variants that may impair detoxification of DNA-reactive chemicals will
support further work to identify gene-environment risk factors for the development of lymphoma in boxers and other dogs.
Student Support: Laboratory of Lauren Trepanier, DVM, PhD
Evaluating body condition index with DEXA and deuterium oxide in big brown bats (Eptesicus fuscus)
Joe Fasig, Sybill Amelon, Sarah Hooper
University of Missouri CVM VRSP, Columbia, MO (Fasig), USDA Forest Service, Columbia, MO (Amelon),
Department of Veterinary Pathology, University of Missouri CVM, Columbia MO (Hooper)
Body condition indices have been developed when assessing subjects in order to numerically score and place subjects into
categories. In bats, body condition index (BCI) is determined by mass to forearm length ratio. Although, BCI can be easily
defined, measuring forearm length can be variable among researchers, creating different scores. The purpose of this study is
to determine body composition measurements in big brown bats (Eptesicus fuscus) by using dual-energy x-ray absorptiometry (DEXA) and deuterium oxide and comparing with given body condition indices. This experiment can also determine
if deuterium oxide can be used as more objective tool in the field. We hypothesized that DEXA and deuterium oxide can
provide an objective measurement of body condition and reduce variability between observers. Big brown bats ranked on a
body condition scale of 1 to 5 with 1 being emaciated to lean and 5 being excessive body fat were anesthetized and measured
for body condition index by dividing mass by forearm length then, scanned with DEXA, and injected with deuterium oxide
to determine rate of water turnover and body condition by evaluating rate of deuterium oxide enrichment decline in water
extracted from blood collected. Preliminary results of the heaviest class of individuals (n=9) mean of all bats forearm 47.33
(SD + 2.37). Mean forearm length between male (n=4) and female (n=5) was 49.39 mm (0.61) for males and 45.78 c=mm
(1.88) for females. Males had a mean body condition score (BCS) of 4.03 (0.71) and a mean percent body fat of 26.5 (1.97).
Females had a mean BCS of 4.05 (0.82) and a mean percent body fat of 28 (8.96).
Research Grant: USDA Forest Service
Student Support: An endowment established by IDEXX-Bioresearch
142
Galya Fedderly, M. Suresh
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin, Madison, Wisconsin
Asthma is a chronic airway disease that affects >300 million people worldwide. In asthmatic individuals, disease exacerbation is often precipitated by respiratory viral infections. Further, infection with respiratory viruses during childhood augments
the risk for developing allergic asthma. However, the mechanistic link between influenza infection and asthma is poorly
understood. Bach2 is a transcription factor that governs multiple aspects of T and B cell responses. Significantly, Bach2 gene
variants are associated with a various immune-mediated diseases including asthma in humans. Therefore, Bach2-deficient
(Bach2-/-) mice could serve as a genetically susceptible model to study interactions between allergic asthma and influenza
A virus (IAV) infection. We hypothesize that prior influenza virus infection of Bach2-deficient mice enhances experimental allergic asthma. Our second hypothesis is that experimentally induced asthma in Bach2-deficient mice accelerates and
exacerbates influenza virus-induced lung disease. We will test these hypotheses by: (1) inducing allergic asthma in wild
type (WT) and Bach2-/- mice by administration on chicken ovalbumin (OVA) followed by infection with IAV; (2) infecting
WT and Bach2-/- mice with IAV followed by allergic asthma induction. We expect that experimental allergic asthma will
compromise viral control and exacerbate the virus-induced inflammatory response in lungs, especially in Bach2-/- mice. We
also predict that prior IAV infection augments the inflammatory response to OVA leading to enhanced airway disease. These
studies will pave the way for mechanistic investigations to elucidate the link between asthma and influenza virus infection.
Research Grant: NIH, UO1AI124299
Development of a qPCR assay for Altered Schaedler Flora colonized mice utilizing the groEL gene
Nicole Ferrero, Gregory Phillips, Alexandra Proctor
Department of Veterinary Microbiology (Ferrero, Phillips, Proctor), College of Veterinary Medicine,
Iowa State University, Ames, Iowa
The mammalian gastrointestinal microbiota is composed of hundreds of bacterial species and species variants that, collectively, vastly outnumber the intestinal cells. The importance of this complex relationship and its effect on host health, immunity and nutrient acquisition is well established through numerous studies. However, the complexity of these microbial
populations makes it difficult to identify mechanisms by which individual bacterial species impact the host. To address this
problem, we are exploiting the altered Schaedler flora (ASF) gnotobiotic mouse to study host-microbe and inter-microbial
interactions within the gut. The ASF was developed in the 1970s and represents a group of eight specific bacterial species
that form a stable community in the murine intestine. The goal of this study is to develop an improved real-time, quantitative PCR (qPCR) assay to measure the abundance of the ASF from the mouse intestine. For this, we have designed qPCR
primers that target the groEL gene. This is a conserved gene found in all members of the ASF community in single copy,
but with sufficient diversity to differentiate between ASF species. We have also developed a synthetic ASF microbiome to
improve the accuracy of quantifying the ASF community. The development of this qPCR assay will be used to better understand how diet, antibiotics, inflammation, infection and other changes that impact the host also influence the number and
localization of the ASF community.
Research Grant: NIH T35 Training Grant 4T35OD012199-14
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Comparison of Contemporary Aqueous and Alcohol-Based Scrub Agents for Rodent Surgical Skin Preparation
Elis Fisk, Jacquelyn Del Valle, F. Claire Hankenson
College of Veterinary Medicine (Fisk, Hankenson), Campus Animal Resources (Del Valle, Hankenson), Michigan State
University, East Lansing, Michigan.
Rodent surgery is a common technique used in comparative medical research. However, methods for surgical site disinfection of rodent skin are variable. Since contemporary husbandry practices result in a relatively clean population of laboratory mice, we questioned whether the more traditional skin preparation technique utilizing three alternating scrubs prior to
incision is needed to achieve skin sterility. Furthermore, by comparing both aqueous and alcohol-based scrub solutions we
aimed to determine which agents are most efficacious in terms of antimicrobial properties, reduction of surgical site contamination, and maintenance of intraoperative body temperature during a surgical procedure (laparotomy). We compared
the performance of sterile saline, 70% ethanol (EtOH), Betadine , Vedco ChlorHex-Q Scrub , Sterillium Fragrance Free
Surgical Rub, 3MTM AvagardTM Surgical Hand Antiseptic, and Purell Instant Hand Sanitizer for rodent skin surgical preparation. Skin was cultured between each round of surgical scrub preparation to assess site contamination. Core temperatures
were recorded every minute throughout the 45 minute procedure, including laparotomy and skin closure. Photographs of
the surgical site were scored for evidence of inflammation following skin shaving, incision site closure, and at 4 and 7 days
post-operatively, prior to euthanasia. A final biopsy of the surgical site was assessed using histology for evidence of tissue
changes. Our preliminary data suggests that waterless alcohol-based solutions are comparable to aqueous-based povidone
iodine and chlorhexidine in prevention of postoperative infection and moderation of hypothermia, while requiring fewer
alternating applications to achieve skin sterility.
Research Grant: Campus Animal Resources Michigan State University Internal Funds
Student Support: College of Veterinary Medicine (Nathan Brewer Endowment)
Nexelom cellometer live cell and viability verification and error evaluation for the canine stem cell assay
Payge B. Fleming, Adrienne B. Cromer, Mark L. Weiss
Department of Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS
Data validation and reproducibility are critical steps in science and concerns about data reproducibility between laboratories
are important. Cellometers are instruments used to count the number of live and dead cells in a sample automatically. The
technology automates the manual hemocytometer method. Currently, no research has been done to validate that counts are
uniform across multiple cellometers, and having such information is valuable for cell product quality assurance and quality
control. Here, we evaluated cell counts performed using two Nexelom Visions and one Nexelom Auto 2000. Three trials
using three different canine stem cell samples were aliquoted and stained four separate times. The samples were then counted using the three cellometers to enumerate reproducibility within and uniformity between systems. Specification from the
instrument manufacturer indicated that the coefficient of variation within each machine should be <10%. The CVs for cell
viability for all machines were <10%. The CVs for live cell counts were not <10%, with the Vision having the highest CV
values overall. The viability values were within 5% of each other on average. In contrast, live cell count CVs, accounting
for cumulative error, averaged 89.6% between the two Visons and 51.2% between the Vision and the Auto. Our goal is to
determine why the differences exist between the Visions and the Auto so that cell counts across machines can be reduced
to within 5% error of each other. If errors larger than 5% are consistent between machines, then the cellular quantifications
cannot be considered comparable and accuracy cannot be validated. This would be a barrier to reproducing results across
laboratories.
Research Grant: Contract with Medivet Biologics
Student Support: NIH T35OD010979
144
Isolation of antibiotic-producing bacteria from New River Valley (NRV) soil
Stephanie Folkerts, Nammalwar Sriranganathan
NC State University CVM, Raleigh, NC (Folkerts) and Department of Biomedical Sciences & Pathobiology, VA-MD
CVM, Blacksburg, VA (Sriranganathan)
The use and misuse of antibiotics in medicine and as growth promoters in livestock production appear to have selected for
drug-resistant bacteria like those in the ESKAPE pathogen group (E. faecium, S. aureus, K. pneumoniae, A. baumannii, P.
aeruginosa, and Enterobacter spp.). As a result, there is a need for the discovery of novel antimicrobials. Four soil samples
from two soil sites were collected from the NRV for analysis of bacteria with antimicrobial activity. Sites were chosen based
on the presence of aquatic and terrestrial soil in close proximity so that activity in these different types of soils could be
compared. Soil dilution plating was used to identify bacterial colonies and these colonies were plated onto agar mixed with
S. aureus and E. faecalis. Bacteria with antimicrobial activity were identified and were further tested using the cross-streak
method. If the bacteria showed inhibition of S. aureus or E. faecalis, their culture supernatants were further evaluated using
paper discs. Aquatic soil samples did not clearly demonstrate more or less activity than terrestrial samples. Eight bacterial
colonies showed strong antimicrobial activity in the original agar plates and on cross-streak plates. One isolate’s supernatant
showed activity on disc diffusion plates. This suggests that most culture supernatants did not have high enough activity to
produce entirely clear zones of inhibition. It may suggest that the antimicrobial production requires an induction process
i.e., presence of another microorganism. Further research should incorporate media which supplies necessary stimuli such
as broth cultures which use dialysis tubing to allow different bacteria’s metabolites to interact with isolates of interest.
Research Grant: NIH grant 4T35OD011887-10
Student Support: NIH grant 4T35OD011887-10
Parvovirus detection by PCR and ISH is associated with myocarditis and cardiomyopathy in young dogs
Jordan Ford, Alex Molesan, Laura McEndaffer, and Kathleen Kelly
Perinatal parvoviral (PV) infection of pups born to naive dams causes necrotizing myocarditis resulting in cardiac failure,
sudden death, and high mortality at 3-4 weeks of age. Acute infections are characterized by the presence of viral inclusions,
necrosis, and colocalization of PV antigen to cardiomyocytes. Viral inclusions and detection of antigen are rapidly lost in
pups surviving acute infection. Given the heart’s stereotypic response to injury, we hypothesized myocarditis or cardiomyopathy in dogs <2years was frequently initiated by perinatal PV infection. For our study, DNA was extracted from archived
formalin-fixed paraffin-embedded (FFPE) tissues from 41 such cases and age-matched controls from the NYS Animal
Health Diagnostic Center archives. Samples were examined histologically and scored for myocardial necrosis, inflammation, and fibrosis by a blinded pathologist. Conventional PCR and sequencing was performed on FFPE tissue to detect the
VP2 region of CPV-2. PV DNA was detected in 12/41 myocarditis/fibrosis cases and 2/41 controls. We found that myocardial PV DNA was significantly associated with myocarditis and/or fibrosis (p=0.007). PCR results were confirmed by in situ
hybridization (ISH) which identified PV DNA in cardiomyocytes of juvenile and young adult dogs (median age 61 days)
and in juveniles with PV enteritis, indicating a longer window of cardiac susceptibility to PV myocarditis than previously
reported. PV DNA was also detected in cardiomyocyte nuclei of dogs with moderate to severe myocardial fibrosis (age
range 126-243 days) suggesting earlier myocardial damage by PV. Given the temporal limitations of antigen-based testing,
ISH offers expanded diagnostic utility for the identification of PV.
Student Support: AVMA-AVMF 2nd Opportunity Research Scholarship
145
Does re-emerging St. Louis encephalitis virus in California show increased fitness compared to older strains?
Samantha Fousse and Lark L. Coffey
Integrative Genetics and Genomics Graduate Group (Fousse) and Department of Pathology, Microbiology and
Immunology (Coffey), School of Veterinary Medicine, University of California, Davis
St. Louis encephalitis virus (SLEV) is a flavivirus that periodically causes severe and sometimes fatal disease in humans.
The last SLEV activity detected in mosquitoes in California prior to 2015 was in 2004. In 2015, there was an SLEV outbreak
in Arizona resulting in one fatality, followed by detection of the virus in Southern California one month later. The circumstances promoting re-emergence of SLEV in California are not known. Emerging strains may have increased their fitness
relative to older strains from California (CA03) and their closest genetic relative, a 2005 strain from Argentina (AR05). The
goal of this project is to compare relative fitness of old and re-emerging SLEV. We hypothesize that a re-emerging 2015
California SLEV strain (CA15) shows augmented in vitro fitness, detected as faster replication and higher titers, compared
to older CA03 and AR05 strains. Replication kinetics of the three SLEV strains were compared using one-step growth
curves generated 0 to 72 hours post-inoculation in duck fibroblasts. Mean SLEV RNA levels were enumerated at each time
point using quantitative real-time reverse transcription polymerase chain reaction. Results were compared using repeated
measures ANOVA with p<0.05 considered statistically significant. Although analyses are in progress, data in support of
our hypothesis will show that the re-emerging CA15 strain displays increased fitness in avian cells compared to CA03 and
AR05 strains. This may allow contemporary SLEV to spread easier among hosts and may have contributed to the recent
re-emergence of SLEV in California.
Research Grant: Start-up funds provided by the Pathology, Microbiology and Immunology Department to Dr. Coffey.
Student Support: NIH T35 Training Grant (UC Davis STAR Program)
Characterization of the pathogenic potential among different species in the spotted rickettsial group
Kristine K. Frady, Pedro Curto, Isaura Simoes, and Juan J. Martinez
Louisiana State University, School of Veterinary Medicine, Baton Rouge, LA (Frady) Department of Pathobiological
Sciences, Louisiana State University, School of Veterinary Medicine, Baton Rouge, LA (Curto, Simoes, Martinez)
Several members the spotted fever group (SFG) rickettsiae are responsible for precipitating severe human and animal tickborne diseases, while other species in this group are non- pathogenic. Members of the SFG are known to survive within
their mammalian host’ endothelial cells. However, recent studies have shown pathogenic species of Rickettsia are also able
to persist within a variety of host phagocytic cells, including monocytes, macrophages and neutrophils. This discovery
suggests that the ability of a rickettsial species to persist inside of a phagocytic cell line may have a major influence on its
pathogenic potential. Here, we evaluated the ability of two SFG Rickettsia species, R. rickettsii (a recognized human pathogen) and R. montanensis (a non-pathogenic species) to survive in murine macrophage cells (RAW) and human endothelial
cells (EAHY926). In order to do this, we cultured the RAW and EAHY926 cells on well plates, inoculated them with their
respective bacteria and then observed the cells at days 1, 3 and 5 of infection via fluorescent microscopy and DNA extraction
followed by qPCR. We predict the pathogenic species, R. rickettsii, will persist in the macrophage cell line (RAW), while
the non-pathogenic species, R. montanensis, will not. The ability of pathogenic rickettsia to surivie within host phagocytic
cells, while non-pathogenic rickettsia cannot, is an interesting phenotypical difference. This difference could possibly lead
to advances in vaccines, treatment plans, and future diagnostic tests for newly isolated strains. What are the genetic determinants that govern growth within these professional phagocytes? This is something the Martinez lab is actively studying.
Research Grant: National Institute Of Allergy And Infectious Diseases of the NIH R01 AI072606
Student Support: National Institutes of Health LSU School of Veterinary Medicine Summer Scholars Program
146
Primary neuron culture autapses as a model for investigating opioid receptor desensitization
Emmanuelle A. Furst, Marissa J. Metz, and Shane T. Hentges
Department of Biomedical Sciences, Colorado State University, Fort Collins, CO
Opioids are the most effective drugs currently available for pain management, but their use for chronic pain comes with the
risk of tolerance and addiction. In order to design better opioid drugs, it is necessary to understand the regulation of opioid
receptor function since adaptive processes such as acute receptor desensitization and internalization may be key in the development of opioid tolerance. Despite the fact that presynaptic and postsynaptic m-opioid receptors (MORs) have the same
known structure and intrinsic properties, the presynaptic MORs appear to be resistant to acute desensitization. The goal of
the present study was to establish a system whereby MORs located in the pre- and postsynaptic neuronal compartments
could be studied in a single neuron. Proopiomelanocortin (POMC) neurons from the arcuate nucleus of the hypothalamus
were chosen as a model cell type because they are regulated both presynaptically and postsynaptically by MORs. Primary cultures of fluorescently-labeled POMC neurons were prepared at low density to encourage the formation of recurrent
synapses (autapses). In cells with autapses, it was possible to use patch-clamp electrophysiology to simultaneously monitor
the ability of MOR agonists to induce a postsynaptic outward potassium current and to inhibit the presynaptic release of
neurotransmitter. If the previously described differential desensitization of the pre- and postsynaptic responses is present in
this autapse preparation it will indicate a compartment-specific mechanism for the resistance to desensitization. The autapse
system will provide a powerful model for future investigations into the mechanisms underlying presynaptic resistance to
acute desensitization.
Research Grant: NIH R01DA032562 and a gift from the Monfort Family Foundation (STH)
Cardiovascular effects of epinephrine on Alligator mississippiensis recovering from isoflurane anesthesia
Nicole Furst, Bonnie Gatson, Simon Swift, and Jim Wellehan
Department of Anesthesia (Gatson), Department of Cardiology (Swift), and Department of Zoological Medicine
(Wellehan), College of Veterinary Medicine, University of Florida, Gainesville, Florida
Crocodilians are commonly anesthetized using inhalant agents during procedures to minimize risk to patients and personnel. However, crocodilians often times are slow to recover from inhalant anesthesia, which delays return to their enclosures
and increases costs to clients. Previous studies indicate that intramuscular epinephrine significantly reduces recovery time
from inhalant anesthesia in American alligators (Alligator mississippiensis). It is conceivable that cardiovascular right to
left shunting, which occurs naturally during underwater submersion, plays a role in delaying anesthetic recovery. However,
it is unknown how volatile anesthetics and epinephrine affect the complex cardiovascular anatomy and shunting patterns of
the crocodilian heart. In this blinded cross-over study, six American alligators underwent isoflurane anesthesia for ninety
minutes. After isoflurane was discontinued, epinephrine (0.1mg/kg) or equal volume of saline was administered intramuscularly. Echocardiography was performed before anesthetic induction, during maintenance of anesthesia, and immediately
following administration of treatment. Physiologic parameters (heart rate and respiratory rate), concentration of airway gases, and recovery parameters were recorded every five minutes during anesthetic recovery. Description of shunting patterns
observed by echocardiography during different stages of anesthesia and recovery will be described. Also, the time from
discontinuing isoflurane to spontaneous ventilation, return of palpebral and withdrawal reflexes, and spontaneous movement
will be compared between treatment groups.
Research Grant: Association of Reptilian and Amphibian Veterinarians Grant, University of Florida College of Veterinary
Medicine intramural funds
147
Marble Burying for Assessing Postoperative Pain in Rats Treated with Meloxicam or Sustained-Release Meloxicam
Kristopher G. Galang and Robert K. Onaga, Jessica D. Ayers, and Lon V. Kendall
Laboratory Animal Resources, Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine
and Biomedical Sciences, Colorado State University, Fort Collins, Colorado
Rats are a common model for research studies involving the use of pain-inducing events that must be ameliorated with analgesics. Rats bury novel objects as a function of their natural behavior to new objects and is a common method to evaluate
anxiety in rats. This study investigates marble burying as a unique means to assess pain in laboratory rats. We hypothesize
that rats in pain will bury less marbles. Male Sprague-Dawley rats were castrated or received anesthesia only and divided
into 3 respective treatment groups of 6 rats each: meloxicam (2 mg/kg SC once daily for 3 days), sustained-release meloxicam (SRM; 4 mg/kg SC once), or saline. Assessments were done at baseline and 1, 6, 12, 24, and 48 h post-surgery. Rats
were observed for 5 minutes to evaluate behavioral indicators of pain including orbital tightening, wound licking, grooming,
and rearing. Then, rats were placed in individual marble-containing cages and given 30 m to bury marbles. Castrated rats
given SRM had the highest rearing activity post-surgery, while castrated rats given meloxicam had the highest grooming
and the least wound licking activities post-surgery. Castrated rats in the saline treated group buried fewer marbles and had
the lowest magnitude increase in marble burying from 1 h to 48 h post-surgery as compared to castrated rats that received
analgesics and the anesthesia-only groups. This suggests the marble burying test may be used as an adjunct to other pain
assessment indicators to identify pain in rats.
Student Support: ASLAP Foundation
Custom 3D-printed drill guides for surgical stabilization of the canine cervical spine - a cadaveric study
Ashley Gavitt, Kristen Malinak, Christopher Mariani, Ola Harrysson, Denis Marcellin-Little
Departments of Clinical Sciences, College of Veterinary Medicine (Mariani, Marcellin-Little), and Industrial and Systems
Engineering, College of Engineering (Harrysson), North Carolina State University (NCSU), Raleigh, NC
The conventional method for surgical stabilization of the canine cervical spine involves placement of metal implants and
polymethylmethacrylate via a freehand technique. Ideal implant placement into the vertebrae is extremely difficult as both
the vertebral arteries and the spinal cord are in close proximity and at risk of injury. The objective of this study was to ascertain if patient-specific 3D-printed drill guides with pre-determined trajectories would result in accuracy and safety acceptable
for clinical use. Computed tomographic (CT) scans of cervical spines from 12 canine cadavers were imported into Mimics
(Materialise) and ideal drill trajectories were planned for vertebrae C4 - C7 for each cadaver. Trajectories were designed
for two transverse process implants and two vertebral body implants for each vertebra. These trajectories were utilized to
design drill guides within 3Matic (Materialise). The custom drill guides and a biomodel of each cadaver spine were printed
on a Projet printer (3D Systems) in VisiJet M2 RCL. The drilling step of the stabilization procedure was simulated on the
biomodels and then performed on cadaver spines followed by repeat CT scanning. Accuracy will be evaluated by measuring
angular and translational deviation between planned and actual trajectories. Safety will be evaluated by a grading scheme
assessing breaches of neurovascular structures. Drill guides and biomodels for 3 cadavers have been printed and drilling
successfully simulated on these models. The cadaver spines have been drilled and the analysis is ongoing. In conclusion,
the creation of custom 3D-printed drill guides for the canine spine is feasible appears accurate based on preliminary studies.
Student Support: NCSU College of Veterinary Medicine, Fund for Discovery, and NIH grant #T35OD011070
148
Refining approaches to assess microbiota and low-profile pathogens in the tick vector, Amblyomma maculatum
Nancy A. Gavron, John V. Stokes, Si Hong Park, Steven C. Ricke, Jung Keun Lee, Kaitlin J. Graham,
Sharon E. Cannaliato, and Andrea S. Varela-Stokes
Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS
(Gavron, Stokes, Lee, Graham, Cannaliato, Varela-Stokes), Division of Agriculture, Department of Food Science,
Center for Food Safety, University of Arkansas, Fayetteville, AR (Park, Ricke)
Amblyomma maculatum, Gulf Coast ticks (GCT), harbor species of bacteria and protozoa, including emerging zoonotic
pathogens, Rickettsia parkeri and Panola Mountain Ehrlichia (PME), as well as the canine protozoal pathogen Hepatozoon
americanum. While infection rates of R. parkeri in GCT populations have been recently documented, little is known about
infection rates of PME and H. americanum. Further, recent studies on tick microbiota are limited; with reports analyzing
extracts not enriched for prokaryotic DNA, and protozoal contributions not considered. Our objectives were to explore the
bacterial microbiome of individual adult GCT, while evaluating infection rates of H. americanum and PME in archived
GCT. We hypothesized that the microbial community of our GCT would include bacterial species previously not detected,
while PME would be rarer than H. americanum. We collected questing adult GCT and extracted DNA using a protocol
that selects for prokaryotic DNA; extracts were submitted for sequencing using an Illumina MiSeq platform, with results
pending. To evaluate infection rates in archived GCT, we first addressed challenges in detection assays. The rarity of PME
prompted production of a plasmid control to be used in a TaqMan quantitative (q)PCR. Initial assays for H. americanum
used published 18S rRNA primers in a nested PCR. We could successfully detect H. americanum in blood but not in ticks
due to significant similarities in the primer sequences to GCT sequences. New primers were designed and utilized in a
SYBR green qPCR. The optimization of these protocols will be used to better understand the infection rate of these pathogens in GCT and evaluate the influence of microbial diversity on pathogen transmission.
Research Grant: NIH COBRE and USDA “Hatch Act” CRIS
miR146a is an endogenous regulator of both hematopoiesis and bone mass
Jennifer A. Geisler, Blake E. Hildreth III, James Lee, Michael C. Ostrowski, Sudu Sharma
College of Veterinary Medicine, The Ohio State University, Columbus, OH (Geisler), Comprehensive Cancer Center,
The Ohio State University, Columbus, OH (Hildreth, Lee, Ostrowski, Sharma)
MicroRNAs (miRNAs) are non-coding RNAs that regulate cellular activity by binding to protein-coding RNAs, suppressing translation or causing RNA degradation. One microRNA, miR-146a, is a key regulator of inflammation, where it is a
physiologic break on immune activity. In the skeleton, bone-resorbing osteoclasts share a common progenitor with macrophages of the myeloid lineage within the hematopoietic heirarchy. miR-146a has been shown to negatively regulate
osteoclast differentiation and function by our laboratory and others in vitro. Because of this, we wanted to investigate the
role of miR-146a in bone biology and hematopoiesis in vivo. We used or designed two transgenic mouse models for this
purpose - a knock-out (KO) mouse where miR-146a has been deleted from the mouse genome and a knock-in (KI) model
where miR-146a is overexpressed in the mouse genome. Male and female mice were aged 6-7 months and full necropsies
and tissue samples were collected for phenotyping. Weights of the spleen and liver (hematopoietic organs) were obtained as
well as skeletons for radiographic and hematopoietic evaluation. In both male and female KO mice and male KI mice there
was a significant increase in spleen weight compared to wild-type controls. Female KO mice had significantly greater liver
weights. These findings suggest changes in the number of cells within the hematopoietic hierarchy. KO mice had decreased
bone density and KI mice had increased density, suggesting that miR-146a is also a negative regulator of osteoclast function
in vivo. These findings indicate that miR-146a regulates both hematopoiesis and bone mass. Further phenotyping is currently underway to provide insight into the role of miR-146a in these processes.
Research Grant: NIH-NIAMS R01AR0447-15, OSU-CCC Pelotonia Post-doctoral fellowship
149
Natasha Ghearing, Jason Stull
Department of Preventative Veterinary Medicine, College of Veterinary Medicine, The Ohio State University,
Columbus, Ohio
Little is known about the husbandry, preventative health practices, and pathogen prevalence within dogs participating in
American Kennel Club (AKC) conformation dog shows. These shows provide cause for concern due to the close interaction
of many dogs from various geographical locations, differing preventative care standards, and infectious disease risks which
may contribute to the spread of pathogens, including those with zoonotic potential. A cross sectional survey was used to
obtain information on current dog husbandry and preventative health practices at AKC shows in Ohio. Along with the surveys, matching dog fecal samples were collected and processed for multidrug resistant ESBL organisms (blaCMY, blaCTX-M,
blaKPC, blaNDM-1), Salmonella spp., and common gastro-intestinal parasites. To date 38 samples have been tested. Results
suggest Salmonella spp. is infrequently identified in show dogs (n=2; both reportedly fed a raw meat diet), however 32%
(n=12) and 38% (n=13) of fecal samples carried bacterium resistant to Cefoxitin, a second generation cephalosporin, and
Nalidixic Acid, a quinolone antibiotic, respectively. No other tested resistance profiles were detected. The findings to-date
suggest high antimicrobial resistance in the show dog population and resulting potential concern for dog and human health.
Research Grant: American Kennel Club and Ohio State University Grants
Student Support: American Kennel Club and Ohio State University Grants
C.L. Gilfeather, G.Yray, J. Nichols, E. Kummari, S. Dhital, and B.L.F. Kaplan
College of Veterinary Medicine, Mississippi State University, Mississippi State, MS
Multiple sclerosis (MS) is an autoimmune disease in which the immune system attacks cells in the central nervous system
(CNS) that provide neuronal myelination. A plant-derived cannabinoid, cannabidiol (CBD), is currently being investigated
to help reduce spasticity in MS patients.With its potential for increased use, it is important to understand the mechanism
by which CBD is effective in MS. We used a mouse model of MS, experimental autoimmune encephalomyelitis (EAE), to
study disease following a 5-day oral treatment with CBD. EAE was induced by injecting mice with a self-peptide, myelin
oligodendrocyte glycoprotein (MOG35-55), which is found in myelin. We hypothesized that CBD would suppress T cells in
EAE. Clinically, CBD delayed initiation of disease by 2 days, although mice at end stage disease exhibited similar clinical
signs to EAE mice. Upon necropsy, splenocytes (SPLC) and draining lymph node (LN) cells were stimulated with MOG
or MOG + anti-CD3/28 beads for 48 hrs. No CBD effect was observed on extracellular production of IFN-g or IL-17A in
either lymphoid organ as assessed by ELISA. There was a slight decrease by CBD in intracellular IFN-g in both CD4+ and
CD8+ T cells in LN as compared to EAE mice as assessed by flow cytometry. Similarly, CBD decreased intracellular IFN-g
in CD4+ T cells in SPLC. Direct analysis of CD25 and CD69 activation markers on CD4 or CD8 cells revealed a slight increase in CD69 in LN cells with disease but no further alteration by CBD. In conclusion, while CBD delayed disease there
was little effect on immune function in SPLC and LN at end-stage disease. The results suggest a longer CBD exposure might
be effective for clinical signs and produce immune suppression in EAE.
Research Grant: Funding Provided by Research Grant P20GM103646
Student Support: Summer Research Program funding from MSU CVM
150
Calculation of body surface area using computed tomography-guided modeling in dogs and cats
Renee E. Girens, Kim A. Selting, Charles M. Maitz, Jimmy C. Lattimer
MU VRSP (Girens) and Department of Veterinary Medicine and Surgery (Selting, Maitz, Lattimer),
College of Veterinary Medicine, University of Missouri, Columbia, MO.
Cancer chemotherapeutic drugs are often administered using an estimation of body surface area (BSA). Increased chemotherapy toxicity may be seen in small dogs using BSA-based dosing, while large dogs may be under-dosed leading to
decreased efficacy. Shape and conformation differences among dogs are vast, and using one uniform shape constant may
be inaccurate. Computed tomography (CT) can accurately determine BSA and could modify the existing BSA formula. Retrospectively, full-body CT scans were used from dogs of varying sizes requiring diagnostic imaging. BSA was determined
using XiO radiation treatment planning software, then compared to the estimated BSA based on weight in kilograms and to
another treatment planning system, RayStation. Paired T-test was used to compare the two computer-based methods. Additional cases from both dogs and cats were collected prospectively to document morphometry and body condition scoring.
Of the 16 retrospective scans, results show discrepancies up to +/- 17% between XiO CT-based and calculated BSA, and
up to 15% using RayStation. Paired T-test showed no significant difference between computer methods, but a significant
difference between computer and formula-based calculations (P=0.008). Prospectively, discrepancies up to 8% have been
found. CT-calculated BSA did not correlate with estimated BSA. This magnitude of error could result in significant over- or
under-dosage of chemotherapy. Adjusting the K constant alone may be inadequate to correct this. These findings will help
refine the existing formula to better estimate the appropriate starting dosage for dogs and cats.
Student Support: an endowment established by IDEXX-BioResearch and Merial, a Sanofi company
Estrogen protects progestin-treated mice against intravaginal transmission of cell-associated HIV-1
Melissa E. Glick, Nirk E. Quispe Calla, Rodolfo D. Vicetti Miguel, Janelle Gabriel, Jesse Kwiek, and Thomas L. Cherpes
College of Veterinary Medicine (Glick), Department of Microbial infection & Immunity, College of Medicine
(Quispe Calla, Vicetti Miguel, Gabriel, Kwiek, Cherpes) and Department of Obstetrics & Gynecology,
College of Medicine (Cherpes), The Ohio State University, Columbus, OH
The progestin-only hormonal contraceptive depot-medroxyprogesterone acetate (DMPA) is commonly used by young women in sub-Saharan Africa and other HIV endemic areas. While clinical data indicates that DMPA use increases risk of HIV
acquisition, the underlying biological mechanism for this association is uncertain. Herein, we sought to define this relationship using humanized mice. Female humanized NOD scid gamma (NSG) mice were reconstituted with human HIV-negative
peripheral blood mononuclear cells (hPBMC). Mice were treated with DMPA or DMPA and intravaginal estrogen cream 5
days before infection. All mice were vaginally inoculated with IL-2 stimulated HIV-1 infected hPBMCs 14 days after reconstitution. At 10 days post infection, mice were euthanized and spleens were harvested and processed to generate single cell
suspensions that were then stimulated with human IL-2 for 8 days. HIV infection status of the cell culture was determined
using a luciferase reporter gene assay in TZM-BL cells. These results showed DMPA treated mice were 100% susceptible to
cell-associated HIV-1 infection, whereas all mice treated with DMPA and estrogen cream were protected against infection.
Vaginal tissues were also processed for expression of a cell adhesion protein, desmoglein-1a (DSG-1a) by RT-PCR. We
found DSG-1a expression was significantly downregulated in mice treated only with DMPA. Together, our results suggest
DMPA increases susceptibility to genital HIV-1 infection by decreasing vaginal epithelial integrity, and estrogen restores
this integrity, protecting DMPA-treated mice from HIV-1 infection. Our results may offer a strategy for providing hormonal
contraception that is less compromising of genital mucosal barrier function.
Research Grant: The Ohio State University College of Medicine
151
Expression and Purification of PrM and E Proteins of Zika Virus in Sf9 Cells
Chiyu Guan, Yonghai Li, Wenjun Ma
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University,
Manhattan, KS.
Zika virus (ZIKV) is brought to our attention after its worldwide case reports. ZIKV belongs to Flaviviridae family and
Flaviviurs genus, with a single positive strand of RNA. ZIKV is mainly transmitted through bites from Aedes aegypti
mosquito. It can cause microcephaly, an untreatable brain defect in fetuses and mostly influenza-like symptoms in adults.
There is no specific treatment or vaccine currently available for ZIKV disease. Previous studies showed that two structural
proteins unique to ZIKV, precursor membrane (prM) and envelop protein (E), interacted with various receptors on the host
cell surface and were most immunogenic. In this project, prM and E proteins were produced via BaculoDirectTM Baculovirus
Expression System in Sf9 cells. Electrophoretic bands for the PCR products of prM and E DNAs showed expected sizes on
the agarose gel, suggesting the primers designed amplified the DNA fragments of interest. Using the same primers bacterial
colonies were verified to contain DNAs of interest after the fragments were transformed into a pENTRTMTOPOR vector. An
expression construct was then generated by performing an LR recombination reaction between the entry clone and BaculoDirectTM Linear DNA and transfected into Sf9 cells. The cells showing CPE were collected and the proteins were purified
by Ni-NTA agarose and verified by Western blot using prM/E protein antisera and anti 6X His-tag, respectively. This suggested the proteins we produced via BaculoDirectTMBaculovirus Expression System in Sf9 cells had sensitive and specific
antigen-antibody reaction with prM/E protein antisera. They can potentially be used as vaccine candidates, diagnostic agents
and for further studies on ZIKV.
Research Grant: College of Veterinary Medicine intramural funds
The use of a B cell tetramer to evaluate the memory immune response to PRRSv
Kevin L. Gustafson, Michael C. Rahe, and Michael P. Murtaugh
Department of Veterinary and Biomedical Sciences, University of Minnesota, St. Paul, MN
Vaccination is critical for preventing and controlling disease is swine herds. It enhances animal welfare and saves producers
resources which would otherwise be needed for treatment following infection. However, testing vaccine efficacy, which is
vital for the development of new vaccines, requires expensive scientific trials involving disease challenge, and is laborious
and slow. A promising alternative is assessing the memory B cell response, since memory B cells mediate immune protection, and can be assessed in simple blood samples at various times following vaccination. B cell tetramers may be used to
track an adaptive memory immune response to a pathogen in times of a disease outbreak. Therefore, we developed a B cell
tetramer reagent using the non-structural protein 7 (nsp7) of porcine reproductive and respiratory syndrome (PRRS) virus
for the detection of PRRSV NSP7 specific memory B cells. The tetramer has a fluorescently labeled complex of four nsp7
molecules linked to one streptavidin through biotinylation. The product was tested by immunoblot and gel electrophoresis
for quality, and tested with specific reactivity with PRRSV-immune serum for functionality. Specific anti-nsp7 antibody-B
cell tetramer complex was isolated using magnetic microbeads and evaluated by staining an SDS-PAGE gel to visualize
the porcine antibodies. This well-characterized tetramer reagent will be used to assess memory B cell responses in pigs to
natural infection and vaccination. Additionally, this method may prove useful in various food animal systems for the characterization of the memory immune response following vaccination or natural exposure to pathogens.
Research Grant: American Association of Swine Veterinarians
Student Support: Funding was provided by Merial Limited and the College of Veterinary Medicine, University of MN
152
Investigating novel small compounds and their effect on inflammation and chondrogenesis in joints
Abigail L. Haffner, Manoel F Neto, C. Samuel Umbaugh, and Marxa Figueiredo
Department of Basic Medical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, Indiana;
Center for Drug Discovery, Purdue University, West Lafayette, Indiana
Osteoarthritis affects over 27 million Americans age 25 and older and is a debilitating disease in which articular cartilage
degradation leads to inflammation and pain in the joint. Medical management has not been very successful, as cartilage
is avascular and has a difficult time regenerating itself. In joints, there are laminin receptors (LR) which bind to PEDF
(pigment epithelial derived factor). When PEDF is bound to a LR, it has anti-inflammatory, pro-chondrogenic and anti-angiogenic characteristics. However, the relatively large size and low stability of PEDF at 258C indicates that smaller, more
stable molecules could be an alternative for therapeutic purposes. We developed seven small compounds that bind to the
PEDF docking zone at the LR, with the goal to mimic the effects of PEDF. In order to test our drugs in a pro-inflammatory environment, THP-1 macrophage cultures were stimulated with lipopolysaccharide. Quantitative real-time polymerase
chain reaction (qPCR) was used to assess up- and down-regulation of inflammatory genes, including IL-1b. Compound 3
(C3) appears to be the best PEDF mimic as it reduced IL-1b expression by ~8-fold. A cell pellet chondrogenesis assay using
adipose-derived stem cells (ASCs) was used to examine the effect of compounds on upregulating cartilage-specific genes
at day 8 of differentiation. In the future, ASC pellets will be harvested at differentiation days 7, 14 and 21 to determine the
timing of cartilage-specific gene expression following treatment with the small compounds. From our results, C3 appears
to be the most promising drug for future development since it is able to reduce expression of pro-inflammatory gene IL-1b
and promote ASCs to express cartilage-specific genes.
Research Grant: Showalter Trust Foundation
Vertical non-transovarial transmission of Rickettsia felis in the cat flea, Ctenocephalides felis
Melena R. Hagstrom, Sean P. Healy, Krit Jirakanwisal, and Kevin R. Macaluso
Cat flea spotted fever is an emerging human disease caused by the obligate intracellular bacterium, Rickettsia felis.
Blood-sucking arthropods carry the pathogen, with the cat flea (Ctenocephalides felis) being the primary vector. Within a cat
flea population, R. felis is maintained horizontally (transmitted between adult fleas through mating or sharing a food source)
and vertically (transmitted transovarially from an infected mother to her offspring). These two methods do not account for
the persistence of R. felis seen in infected wild cat flea colonies. Previous studies exhibited a horizontal transmission rate of
3.3-40% and a vertical transmission rate of 2.5% by the 12th generation of fleas. These findings fail to culminate in a 100%
transmission rate which would be required to maintain R. felis in a cat flea population as observed in the wild. On the basis
of this fact, we hypothesized that R. felis could be introduced to flea larvae through injection and that the infection could be
maintained through two molts, pupation, and into adulthood. In order to assess these hypotheses, two replicates of 132 flea
larvae were injected with a suspension of R. felis and the larvae were allowed to pupate and emerge as adults. Quantitative
qPCR showed the presence of R. felis in the adult fleas that emerged from the experiment. Eggs produced by the emerged
adults were assessed via an immunofluorescence assay. These findings reveal a novel route by which R. felis may be maintained in wild cat flea colonies.
Research Grant: NIH R01 AI122672
Student Support: NIH R01 AI122672
153
Characterization of the spectrum of activity and mechanism of a pH-selective inhibitor of M. tuberculosis
Elizabeth R. Haiderer, Chris J. Colvin, Ben K. Johnson, and Robert B. Abramovitch
Department of Microbiology and Molecular Genetics, College of Natural Science, Michigan State University,
East Lansing, MI
PH036 was isolated as a pH-dependent inhibitor of Mycobacterium tuberculosis (Mtb). It was found ineffective in any condition towards Mycobacterium smegmatis (Msm), a non-pathogenic model organism for Mtb. It has been found that PH036
induces reductive stress in Mtb. To determine the spectrum of activity of PH036, half maximal effective concentrations
(EC50) were determined using diverse strains of bacteria: Pseudomonas aeruginosa, Escherichia coli, Proteus vulgaris,
Staphylococcus aureus, and Enterococcus faecalis. The Gram positive strains were found to be more susceptible to PH036
than the Gram negative strains. To explore the mechanism of action of PH036, the oxidation status of Msm exposed to
PH036 was monitored using redox-sensitive green fluorescent proteins (roGFPs). Msm exhibited a more reduced cytoplasm
in acidic conditions, which was enhanced by the presence of PH036 in a dose-dependent manner.
Research Grant: NIH NIAID (R01-AI116605)
A porcine model of vascular cognitive impairment for dementia prevention
Kimberly D. Haight, Erin E. Kaiser, Madelaine N. Wendzik, Simon R. Platt, David C. Hess, Franklin D. West
Regenerative Bioscience Center, College of Veterinary Medicine, University of Georgia, Athens, GA
Dementia is a major public health concern that develops from white matter damage to the brain that is linked to poor blood
supply. Vascular cognitive impairment (VCI) includes anything on the scale from mild cognitive dysfunction to severe
vascular dementia. Currently, there is no effective treatment to prevent dementia in people. There are preliminary results on
a safe and inexpensive treatment termed remote ischemic conditioning (RIC) that has successfully prevented white matter
damage and cognitive impairment in a rat model of VCI. However, due to the differences between a rat and a human brain
in terms of composition and size, there is a need to test this treatment in a large animal model. The aim of this study was to
design and implement a chronic hypoperfusion model in the pig that can accurately model VCI in humans. Surgeries were
done on 4 adult male pigs to constrict the common carotid artery by placing ameroid constrictors around the artery to reduce
cerebral blood flow over time and induce white matter damage. To determine whether this was an accurate model of VCI in
the pig, white matter damage, motor function, and behavioral deficits were tested. MRI was done to assess the changes in
the cerebral blood flow. Gait analysis was done to test for motor function by measuring parameters such as velocity, stride
length, stance percentage, and total pressure index. Open field testing and object recognition tests were done to test for
behavioral changes over time. The results of these assessments showed that the pigs were successfully given VCI. With an
accurate large animal model of VCI established, treatments such as RIC can now be tested in this model in order to optimize
the treatment for future human testing.
Research Grant: UGA-GRU Seed Grant Program
154
Reproductive performance applied to genetic merit and increasing insemination rate of anovular Holsteins
Joseph Hammes, Bobwealth Omontese, Laisa Garcia, Augusto Vilela, Rafael Bisinotto
Department of Veterinary Population Medicine - College of Veterinary Medicine, University of Minnesota
Reproductive efficiency is vital for successful dairy farms. Genomic markers for fertility traits have become increasingly
available to producers, though specific biological events identified by such markers remain largely unexplored. We evaluated pregnancy per artificial insemination (P/AI) and risk of pregnancy loss between d35 and d80 after first AI in Holsteins
(n=372). Cows were classified into quartiles based on genomic predicted transmitting ability (GPTA) for daughter pregnancy rate (DPR), sire stillbirth (SSB) and daughter stillbirth (DSB). Effects of DPR, DSB, and SSB on P/AI and pregnancy
loss were evaluated by logistic regression (proc GLIMMIX, SAS 9.4). The top 25% for each trait (Q4 for DPR and Q1 for
SSB and DSB) was compared with remaining cows by using a contrast statement. Overall P/AI and pregnancy loss were not
affected (P>0.10) by DPR, DSB, or SSB. Top 25% cows for DPR had greater P/AI on d35 (Q4=50.5 vs. Q3=37.1, Q2=41.5,
Q1=42.7%; P=0.09) and 80 (Q4=49.5 vs. Q3=33.7, Q2=37.0, Q1=35.8%; P=0.02). Pregnancy loss was smaller (P=0.10)
for Q4 compared with Q3, Q2, and Q1 (2.1, 8.8, 10.5, and 15.0%, respectively). Because anovular cows are less likely to
be detected in estrus, we are evaluating an alternate program to anticipate first AI postpartum in anovular cows, increasing
overall insemination rate. Cows without CL at the end of a presynchronization (presynch) series (PGF2a at 3663, 5063d)
will be assigned randomly into one of two groups, long and short. Short cows will receive 2 CIDRs and begin a 5d timed AI
(TAI) program on that day. Long and with CL cows will begin a TAI program 12d after the presynch. This ongoing study
will serve as another tool in the assessment of anovular dairy cattle.
Research Grant: Zoetis, Florham Park, NJ - Kamar, Zionsville, IN
Student Support: Department of Veterinary Population Medicine - University of Minnesota - CVM
Compromising X-linked gene silencing in mature B cells through deletion of the long noncoding RNA Xist
Steven Hanes, Camille Syrett, and Montserrat Anguera
University of Pennsylvania SVM, Philadelphia, PA
Females have a greater immunological advantage than men, yet they are more prone to autoimmune disorders. The basis
for this sex bias lies in the X-chromosome, which contains many immunity-related genes. Systemic Lupus Erythematosus
(SLE) is one autoimmune disorder with a strong female bias (~85%), where the number of X-chromosomes increases risk
for SLE development. Females use X-chromosome inactivation (XCI) to transcriptionally silence one X-chromosome,
which equalizes X-linked gene expression between sexes. XCI is initiated during female embryonic development by the
long noncoding RNA XIST (Xist in murine models), which recruits heterochromatin modifiers to form the inactive X (Xi).
After initiation, XCI enters the maintenance phase, where XIST associates closely with the Xi after each cell division to
support gene repression. Heterochromatic modifications and DNA methylation also remain on the inactive X, forming a
memory of transcriptional silencing. Recently, we have found that XCI is maintained differently in B cells compared to
other female somatic cells. Naive mature B cells lack Xist RNA and heterochromatin enrichment on Xi, and these marks
return after stimulation. My hypothesis is that Xist deletion will partially reactivate Xi in splenic B cells, resulting in overexpression of immunity-related X-linked genes. Through use of female wildtype, heterozygous, and homozygous mice for
Xist deletion, and analysis methods like RNA FISH, immunofluorescence, and RNA sequencing, we will determine the
requirement for Xist RNA to maintain heterochromatin enrichment on Xi and the level of gene expression from Xi.
Student Support: NIH-Merial Summer Research Program
155
Effect of a Western-style or a high fat coconut oil diet on metabolism and adipose tissue steroidogenesis
Amanda Hanzel, Cassandra Skenandore, Corinne Bromfield, Romana Nowak, Rebecca Krisher,
Annie E Newell-Fugate
Texas A&M University, CVM, Dept. of Veterinary Physiology and Pharmacology (Hanzel, Skenandore, Newell-Fugate),
College Station, TX; University of Illinois at Urbana-Champaign, AACUP (Bromfield), Urbana, IL; University of Illinois
at Urbana-Champaign, College of ACES, Dept. of Animal Sciences (Nowak), Urbana, IL; National Foundation
for Fertility Research (Krisher), Lone Tree, CO
Seventy percent of American women are overweight or obese which puts them at increased risk for diabetes and elevated
serum androgen levels. Increased androgen levels in these patients may stem from abnormal ovarian or adipose steroidogenesis. Coconut oil has been touted as beneficial for weight loss but there has been little research to support claims of its ability
to modulate metabolism. We hypothesized that female pigs fed a high fat diet containing virgin coconut oil would demonstrate improved glucose homeostasis, have decreased circulating and adipose tissue long chain fatty acids, and have lower
adipose tissue androgen concentrations than female pigs fed a high fat diet containing hydrogenated soy bean oil (Western
Style Diet; WSD). We fed female Ossabaw pigs 2200 kcal of a corn and soy meal diet (control; C; n=11), 6000 kcal of WSD
(n=9), or coconut oil high fat diet (COC; n=3) for a period of 8 months. Blood was collected at standing heat for 7 estrous
cycles and was assessed for glucose, insulin, and free fatty acids. Subcutaneous and visceral adipose tissue was collected at
euthanasia for assessment of testosterone and androstenedione concentrations. Control and COC groups had lower fasting
glucose than the WSD group (p<0.050; C: 64 6 1.5 mg/dL; COC: 66.2 6 1.7 mg/dL; WSD: 80.9 6 5.1 mg/dL). Control
pigs weighed less and had smaller morphometric measurements compared to COC or WSD groups (p<0.05). There was
no difference in subcutaneous adipose tissue testosterone between groups (p=0.13). In conclusion, we find that, despite inducing obesity, a high fat diet containing coconut oil modulates glucose homeostasis similar to a lean diet, but has minimal
impact on adipose tissue androgen concentrations.
Research Grant: NIH K01 RR031274-01A21
Jennifer Anne Hardy, Brian H. Herrin, Susan E. Little, Todd C. Holbrook
Department of Pathobiology, College of Veterinary Health Sciences, Oklahoma State University, Stillwater, OK.
Ticks and tick-borne pathogens are commonly recognized in many veterinary species, but there is currently limited serologic data describing equine infection with Ehrlichia spp. and no true Ehrlichia spp. is known to cause clinical disease in
horses. Geographic analysis of the distribution of horses with antibodies to Ehrlichia spp. is key to describing the most likely tick vector or vectors responsible for transmitting these organisms, and therefore, can provide information about which
Ehrlichia spp. may infect horses. To determine the prevalence of horses exposed to Ehrlichia spp., fifty serum samples were
obtained from each of five geographically distinct diagnostic labs in the United States (GA, OK, PA, TX and WI) (n=250)
and tested by indirect immunofluorescence antibody assay (IFA) to assess the presence of antibodies reactive to Ehrlichia
spp. Of the horses sampled, 13.2% (33/250) had antibodies reactive to E. chaffeensis by IFA. There were significantly more
seropositive horses in Oklahoma, 14/50 (28%), than Georgia or Wisconsin, 4/50 (8%) and 2/50 (4%) respectively. Antibodies were identified in at least one horse from each geographic region, but regions known to support higher populations
of Amblyomma americanum, the known vector of several human and canine Ehrlichia spp., had more seropositive horses
than regions where the tick is less common. These data show that there is a broad distribution of horses with antibodies to
Ehrlichia spp., thus, further sampling is needed to accurately determine the true prevalence of horses exposed to Ehrlichial
agents throughout the US and further describe the likely tick vectors responsible for transmitting the infections to horses.
Research Grant: Krull-Ewing Endowed Professorship in Veterinary Parasitology, June Jacobs Endowed Professorship in
Veterinary Clinical Sciences, OSU CVHS Research Advisory Council
156
Targeting human breast cancer using a radiolabeled cyclic RGD peptide
Jessie Hargis and J. Michael Mathis
Louisiana State University School of Veterinary Medicine (Hargis) and Department of Comparative Biomedical Sciences,
Louisiana State University School of Veterinary Medicine, Baton Rouge, Louisiana (Mathis)
Breast cancer is the second leading cause of cancer deaths among US women. Despite advances detection methods, the mortality rate has remained relatively unchanged over the past several decades. There is an urgent need to identify innovative
alternatives for early detection and therapy. Many breast tumors express a family of surface integrin receptors involved in
tumor metastasis, tumor induced angiogenesis, restenosis, osteoporosis and inflammatory processes. Peptides containing
Arginine-Glycine-Aspartic Acid (RGD) have been shown to bind and inhibit integrin receptors. To test the ability to bind
breast tumor cells in vitro and in vivo for detection, an RGD-containing peptide was commercially synthesized and radiolabeled with Tc99m using EDDA as a co-ligand, then purified using a carbon-18 (C18) spin column. The peptide was examined for specificity in several in vitro assays. Harvested human breast cancer cells (MDA-MB-231-luc) where incubated at
room temperature with different amounts of unlabeled RGD, unlabeled RGE (a control peptide), labeled RGD and labeled
RGE. Samples were washed and radioactivity was recorded. There was no difference between sample groups. An additional
assay was performed on adhered cells, using an endogenous competitor, fibronectin and a synthetic fibronectin peptide,
in varying concentrations. Results of this assay were inconclusive. Immunodeficient mice (Nu/Nu) were also inoculated
with MDA-MB-231-luc cells and tumors were allowed to grow. The purified peptide will be injected IV and one hour post
injection, mice will be imaged by Single Photon Emission Computed Tomography (SPECT). Images will be evaluated for
localization of radioactivity as an indicator of RGD binding and tumor uptake.
Research Grant: LSU Biomedical Collaborative Research Program
Student Support: National Institute of Health
Optimization of a europium-based cytotoxicity assay for equine mesenchymal stem cells
Elizabeth A. Harris, Alix K. Berglund, Lauren V. Schnabel
College of Veterinary Medicine, North Carolina State University, Raleigh, NC
Previous studies have shown that donor equine MHC-mismatched mesenchymal stem cells (MSCs) incite both a recipient
cell-mediated and humoral immune response. In addition to concerns for recipient safety, these findings lead to concerns
over donor MSC clinical efficacy if the cells are targeted for death before exerting their maximal beneficial effects on healing. Recent studies have shown that Transforming Growth Factor-b2 (TGF-b2) treatment is able to significantly downregulate MHC expression on the surface of MSCs. Studies are now needed to determine if this reduced MHC expression results
in reduced cell-mediated cytotoxicity. Although Chromium(Cr)-51 release has traditionally been used in cytotoxicity assays
to evaluate target cell death, the radioactive isotope poses human safety concerns. A newer assay using Europium (Eu) has
demonstrated higher sensitivity than Cr51 for certain applications without the associated risk. The objective of this study,
therefore, was to optimize a protocol to evaluate MSC cytotoxicity using Eu-based labeling. MSCs were loaded with the
fluorescence ligand, BATDA and incubated in the presence or absence of lysis buffer. Next, Eu solution was added to form
the stable fluorescent chelate, EuTDA, used to indicate cytotoxicity by time-resolved fluorometry. Background signal was
successfully eliminated and spontaneous release minimized through addition of stabilizing agents. Unfortunately, the standard protocol resulted in a consistently low fluorescent signal for lysed MSCs independent of MSC concentration, indicating
that the assay requires further optimization for this cell type. Current efforts are focused on adjusting loading conditions and
plate reader settings for signal maximization.
Research Grant: NIH 1K08AR060875 (Schnabel), MAF D16EQ-405 (Berglund)
Student Support: North Carolina State University College of Veterinary Medicine, Fund for Discovery
157
Examination of Pseudoloma neurophilia infection in the central nervous system of zebrafish (Danio rerio)
Katharine E. Hausmann, Kevin A. Lanham, and Michael R. Taylor
School of Veterinary Medicine (Hausmann), University of Wisconsin-Madison, Madison, WI; Pharmaceutical Sciences
Division (Hausmann, Lanham, Taylor), School of Pharmacy, University of Wisconsin-Madison, Madison, WI
Pseudoloma neurophilia is the most common zebrafish parasite that affects greater than 74% of zebrafish facilities worldwide and currently there are no effective treatments. This microsporidian infects the central nervous system of the zebrafish
and has the potential to influence the outcome and interpretation of studies being conducted on the fish. We developed and
optimized new P. neurophilia DNA primers for polymerase chain reaction (PCR) and confirmed an infection in the brains
and spinal cords in some fish displaying clinical signs. Recent studies have suggested that the spores of the P. neurophilia
are autofluorescent and are transmitted through fecal material. In our experiments, individual fish and dissected tissues were
examined for fluorescent nodules and digested to obtain P. neurophilia DNA. Unlike these previous studies, we found no
fluorescent nodules in infected tissues using both the EGFP and mCherry filters. We next obtained fecal material from both
infected and uninfected fish and were analyzed for traces of P. neurophilia DNA using PCR. Interestingly, we found no P.
neurophilia DNA across 13 samples of fish fecal material and 12 samples of detritus from two separate sources. Our experiment is the first to specifically examine fecal material as a source for horizontal transmission and demonstrates that this is
likely not the source of infection. Due to this finding, we hypothesize that P. neurophilia DNA will be found in sperm, eggs,
or both, demonstrating vertical transmission. To test this hypothesis, we plan to collect both sperm and eggs from live fish
and screen for P. neurophilia DNA by PCR. Our studies will potential reveal the source of infection and provide insights
into treatment strategies.
Research Grant: University of Wisconsin-Madison, School of Pharmacy (Startup Funds)
Perinatal growth hormone imprinting of hepatic cytochrome p450 expression in female rats
Allison M. Hayes, Sarmistha Banerjee, Bernard H. Shapiro
Department of Biomedical Sciences, University of Pennsylvania, School of Veterinary Medicine, Philadelphia, PA
Hormonal imprinting is the event when a developing receptor responds to its target hormone for the first time during a
critical period that permanently establishes its response. It is known that androgens masculinize the brain and reproductive tract, however imprinting has yet to be found in females. Previous studies have shown that hepatic cytochrome p450
monooxygenases (CYPs) are apparently imprinted by growth hormone (GH) in male mammals. The degree of variation
of drug metabolism between sexes is due to the many isoforms of sex dependent cytochrome p450 enzymes. The hepatic
CYPs expression is regulated by the sexually dimorphic secretory GH profile. The male secretory profile is characterized
as episodic where there are periods of time with no detectable amount of hormone and the female profile is continuous. The
objective of this research is to determine whether the female hepatic CYPs are also irreversibly imprinted by GH. Newborn
female rats were injected with monosodium glutamate (MSG), a total GH blocker, GH, MSG+GH, or vehicle. After 120
days, the neonatally treated adult female rats were renaturalized with the feminine GH profile. We analyzed the protein and
mRNA levels of female specific hepatic CYP2C12 expression as well as other isoforms. The results suggest that the sexually
dimorphic expression of the female CYP is irreversibly imprinted shortly after birth by GH demonstrating for the first time
that a sexually dimorphic characteristic needs to be imprinted by the same hormone for both sexes. Molecules of the signal
transduction pathway will next be analyzed to differentiate between the possible imprinted pathways for each sex.
Research Grant: U.S. National Institutes of Health, Eunice Kennedy Shriver National Institute of Health and Human Development (grant HD-061285)
Student Support: NIH/ Merial Grant
158
Identifying antimicrobial resistance patterns in Salmonella enterica Typhimurium from human and cattle sources
Sydney R. Hayter, David R. Smith
Department of Pathobiology and Population Medicine (Smith), College of Veterinary Medicine, Mississippi State
University, Starkville, MS.
Antimicrobial resistance is an issue important to the health of humans, companion and food-producing animals. These
concerns affect public health policy. The National Antimicrobial Resistance Monitoring System (NARMS) is a collaborative effort between the CDC, USDA, and FDA to collect and report data regarding antimicrobial resistance in foodborne
pathogens isolated from human clinical cases, food-producing animals, and retail meat samples. The objective of this study
was to determine the pattern of antimicrobial resistance among Salmonella enterica serotype Typhimurium among human
clinical samples and beef carcass samples from the NARMS datasets. Fifteen years of data (1998-2013) were analyzed to
determine the effects of source (human or beef carcass) and year on the probability of reporting S. Typhimurium resistant
to tetracycline, ceftiofur, gentamicin, ciprofloxacin, multidrug resistance (MDR) as defined by CDC, or an MDR pattern
known as ACSSuT. Data were analyzed for linear trend over time using the logistic regression and the Chi square test for
trend. Significant interactions between source and time existed for MDR (p<0.0001), tetracycline (p<0.0001), and ceftiofur
(p=0.0014). Tetracycline (OR = 0.92, p<0.0001) and MDR (OR = 0.9, p<0.0001) resistance from human samples decreased
over time. ACSSuT resistance significantly decreased over time (OR = 0.92, p=0.0071). The probability to recover ceftiofur-resistant samples from cattle increased (OR = 1.1, p<0.0001) over time. No significant relationships were detected for
gentamicin or ciprofloxacin resistance. The probability of isolating resistant S. Typhimurium was dependent on the time and
source of sampling, and differed among drugs of public health importance.
Research Grant: The Beef Cattle Population Health and Reproduction Program at Mississippi State University. Supported
by the Mikell and Mary Cheek Hall Davis Endowment for Beef Cattle Health and Reproduction
Genome-wide association study seeks connection between stifle morphology and cranial cruciate ligament disease
Eleni Healey, Jessica Hayward, Adam Boyko, and Rory Todhunter
Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY
Cranial cruciate ligament disease (CCLD) is a widespread canine orthopedic condition. Rupture of the cranial cruciate
ligament generates instability in the stifle joint during ambulation, leading to chronic pain, lameness and osteoarthritis. The
evident heritability of CCLD has inspired efforts to characterize genomic regions where susceptibility or resistance may be
encoded. In this study we seek to identify genetic influences on stifle morphology which may affect expression of CCLD
using a genome-wide association study (GWAS). 174 dogs representing 60 breeds had stifle radiographs taken at the Cornell
University Hospital for Animals and were genotyped on the Illumina version 2 High Density mapping array by the Cornell
BioBank. Clinical diagnosis of CCLD for cases (n=161) and controls (n=13) was confirmed by clinical evaluation and
radiographic analysis. We performed 9 quantitative measurements on the stifle radiographs for each dog. Principal component analysis (PCA) of these measurements enabled us to describe the size and shape of stifle joint features. Some of these
features reflect CCLD. PC1 concerned average stifle size. PC2 concerned primarily the tibial plateau angle, also correlated
with the infrapatellar fat pad and tibial tuberosity length. We will conduct a GWAS on both the raw measurements and the
first 4 PCs to identify genetic loci associated with stifle shape which may lead to loci associated with CCLD. If we find any
genomic regions of interest, in-depth analysis of potential candidate genes will be warranted. While the power of the study
is limited by population size and the clinical nature of the data, we hope to provide some insight into the genetic factors
influencing stifle morphology and CCLD.
Research Grant: Genotyping was funded previously by a grant from Zoetis Inc.
Student Support: Student support provided by Merial Foundation.
159
Commercialization of an ELISA for Ornithobacterium rhinotracheale using hemolytic and non-hemolytic isolates
Kari Hecker, N.P. Evans, and F.W. Pierson
Department of Population Health Sciences, Virginia-Maryland College of Veterinary Medicine, Virginia Tech,
Blacksburg, VA
Ornithobacterium rhinotracheale (ORT) is a gram negative, rod shaped, non-zoonotic bacterium causing respiratory disease
in poultry. Our laboratory was previously able to isolate a strain of ORT displaying a-hemolysis in turkeys. This a-hemolytic isolate was used in conjunction with a traditional non-hemolytic isolate to enhance antibody binding in an enzyme-linked
immunosorbent assay (ELISA), which had been standardized and optimized in our laboratory. It was determined through
ANOVA and Tukey’s HSD test that coating plates with previously frozen hemolytic and non-hemolytic antigens mixed
together led to significantly higher optical density (OD) readings for ORT-positive samples. This data was used for the commercialization and scale-up of ELISA plates to determine conditions that give accurate results while extending shelf life.
Plates were tested for viability in various storage conditions (dried or stored wet after being rinsed, as well as being stored at
freezer (-208C) or refrigerator (-48C) temperatures) over a week period. Data through the first four days displayed significant
differences between permutations (average + OD: 0.9737 refrigerator/1.0756 freezer (1 day), 1.0978 refrigerator/1.2281
freezer (2 days), 1.1417 refrigerator/1.0476 freezer (4 days)), with freezer mean values fluctuating. Average negative sample
values for both freezer and refrigerator plates increased through the first four days (27.77% freezer, 38.87% refrigerator)
although the difference between temperatures on a given day was not significant (p>0.05). This indicates that OD readings
increase for both positive and negative samples regardless of storage conditions, but after extended periods of time plates
would lose accuracy.
Bruce W. Christensen, Stuart Meyers, Lysanne Hendrikx
Christensen: Department of Population Health & Reproduction, University of California, Davis Meyers: Department of
Anatomy, Physiology & Cell Biology, University of California, Davis Hendrikx: student faculteit Diergeneeskunde,
Universiteit Utrecht, Utrecht
Artificial insemination using cooled, shipped semen is an integral part of the equine breeding industry. Over time, sperm
produce reactive oxygen species (ROS). ROS interferes with sperm mitochondria membranes. Since mitochondria produces
ATP, damage to mitochondria results in a decrease in sperm motility. Therefore, ROS production is responsible for functional impairments associated with infertility. The use of semen extenders, coupled with storing semen at cooled temperatures,
attempts to minimize the effects of oxidative stress and prolong the life span of semen. The aim of this study is to determine
whether cooling and the use of semen extenders are protective against oxidative stress, and therefore result in improved
sperm motility, over time. Two ejaculates each from 5 different stallions will be collected. Each ejaculate will be divided
into 4 aliquots: INRA 96 (a semen extender) at 25o C, INRA 96 at 9o C, BWW w/PVA (a phosphate salt buffer) at 25o C
and BWW w/PVA at 9o C. All aliquots will be checked for motility, morphology and oxidative stress at 0, 24, and 48 hours.
Motility and morphology will be measured and recorded using a computer assisted semen analysis system, Oxidative stress
will be measured by flow cytometry using dihydroethidium (DHE) and sytox. The expectation would be that oxidative stress
would increase and motility and normal morphology will decrease at 24 and 48 h. An increase over time in oxidative stress,
correlated with a decrease in sperm motility and/or normal morphology, in non-extended or room-temperature samples as
compared to extended and chilled samples will indicate the necessity of using semen extenders, such as INRA 96, and chilling in maintaining higher viability in semen.
Research Grant: Research Grant: Dr. Christensen discretionary funds
Student Support: Student Support: Merial Veterinary Scholar Program 2016
160
Identification and characterization of Yersinia ruckeri from farm-raised catfish in Mississippi
Madeleine V. Hendrix, Stephen R. Reichley, Lester H. Khoo, Patricia S. Gaunt, David J. Wise, and Matt J. Griffin
Thad Cochran National Warmwater Aquaculture Center, College of Veterinary Medicine, Mississippi State University,
Stoneville, MS
Yersinia ruckeri is the causative agent of enteric redmouth disease in salmonid fish. Although most commonly isolated
from trout, there have been limited documented cases in farm-raised catfish. In spring of 2016, Y. ruckeri was isolated from
diseased channel (Ictalurus punctatus) x blue (I. furcatus) hybrid catfish from multiple ponds on a single operation. It is
unknown if this isolation indicates a potentially emerging pathogen or a fluke occurrence. Farm-raised catfish is the largest food fish aquaculture industry in the United States. Given the economic importance to Mississippi and its neighboring
states, it is critical the susceptibility of catfish to this pathogen be evaluated. To this end, channel and hybrid catfish were
challenged with Y. ruckeri by intraperitoneal injection as well as bath immersion to fulfill Koch’s postulates and determine
the relative virulence of Y. ruckeri in catfish. Results suggest hybrids may be more susceptible to Y. ruckeri. Interestingly,
under the conditions used in this study the bacterium did not cause disease by immersion, even under experimentally induced oxygen stress, suggesting the pathogenicity in catfish may be limited. Additional studies at lower temperature are
warranted. Furthermore, Y. ruckeri isolates from both catfish and trout were subjected to phenotypic and genetic analyses
using commercial phenotypic test kits (BBL, Biolog, API20E), motility and TSI slants, estimations of G+C content, gyrB
sequencing, plasmid characterization, and repetitive extragenic palindromic PCR (rep-PCR). These analyses indicate genotypic and phenotypic variations exist between isolates from catfish and trout, although the biological implications of these
differences are presently unknown.
Role of the gerQ determinant in exosporium structure in Bacillus anthracis.
Chris M. Henson, Hsin-yeh Hsieh, George C. Stewart
Department of Veterinary Pathobiology and Bond Life Sciences Center, University of Missouri.
Bacillus anthracis is a gram-positive, spore forming bacterium that is the etiologic agent of anthrax. This zoonotic pathogen is associated with high levels of mortality in infected ruminants and humans with the pulmonary form of the disease.
When nutrients are sparse, such as in the soil, the bacteria will undergo the process of sporulation, to produce the dormant,
specialized structure, the spore. The spore is important as it is the infectious form of the bacterium. The outermost layer of
the spore is the exosporium, which consists of a basal layer and a hairlike glycoprotein nap layer. The exosporium is the
structure which directly interacts with the host innate immune system during the initial stages of the infection. The protein
composition of the exosporium is not well understood. Some of the proteins have been shown to have an effect on exosporium development, such as BclA (the prominent nap layer glycoprotein), BclB (found just below the nap layer), and BxpB
(a basal layer protein). Therefore, we are striving to learn more about which proteins are integral in the formation and anchoring of the exosporium. Previous studies performed with Bacillus cereus indicated that mutants lacking the GerQ protein
had a more fragile exosporium. We used allele replacement mutagenesis to create a B. anthracis bacterium that lacks the
GerQ protein, and purified spores from this strain. Using a Western Blot approach with antibodies targeting known exosporium proteins, we are investigating whether the loss of the GerQ protein affects structural stability of the exosporium in B.
anthracis strain Sterne spores.
Research Grant: The anthrax research is supported by grant R21AI112725 from the National Institute of Allergy and
Infectious Diseases.
Student Support: Supported by the Department of Veterinary Pathobiology
161
Role of DDR1 in revascularization and repair after myocardial cryoinfarction in mice
Emily Herrold, Frank Lee, Shaun Reining, Jane Lee, Megan Jibilian, and Michael Kotlikoff
Myocardial infarction is a major cause of disease in humans in the developed world. Previous studies in this lab have shown
that revascularization and receptor tyrosine kinase activity are important components to myocardial repair and recovery of
cardiac function. While c-kit, a Receptor Tyrosine Kinase (RTK), is known to play a role in revascularization and myocardial repair, other RTKs may be involved. This study further investigates the role of one particular RTK, the discoidin
domain receptor 1 gene (DDR1), in recovery after cryoinfarction in mice. DDR1 null mice that also carry the c-kitBAC-EGFP
transgene were surgically cryoinfarcted. Infarcted hearts from the null and wildtype littermates were analyzed for cardiac
function using echocardiography, revascularization using immunohistochemistry staining with anti-CD31 antibodies, c-kit
activation using anti-GFP antibodies, and infarct size using sirius red and trichrome stainings. In this poster, we present the
findings based on echocardiography, immunohistochemistry staining, sirius red and trichrome staining.
Research Grant: National Heart Lung and Blood Institute 5R01HL127219-02 Vascular Precursors and Cell-Cell Signaling
in Heart Vasculogenesis
Student Support: National Institutes of Health T35 Training Grant 5T35OD010941
Alterations in hepatic glucose metabolism following direct exposure to organochlorine pesticide metabolites
Julie Holdridge, Sandeep Kondakala, Margaret Davis, and George Howell III
Department of Basic Sciences, College of Veterinary Medicine, Mississippi State University, Mississippi State, MS 39762
Diabetes mellitus is an increasingly prevalent disease in both in the United States and worldwide with type II diabetes mellitus, characterized by hyperglycemia driven by insulin resistance, accounting for 90-95% of cases. Although a number of
risk factors for type II diabetes are known, there is evidence that exposure to certain organochlorine pesticides are associated
with the development of type II diabetes as well. Previous studies have demonstrated exposure to p,p’-dichlorodiphenyldichloroethylene (DDE), one of the most prevalent organochlorine compounds, promotes pathological alterations in glucose
homeostasis. However, the underlying cellular mechanisms governing these effects remains poorly understood. The goal of
the present study was to determine if direct exposure to the organochlorine compounds DDE and trans-nonachlor alters glucose homeostasis in the liver utilizing rat primary hepatocytes as an “ex vivo” model of hepatic function. To assess effects
on glucose homeostasis, glucose production, glucose uptake, and glycogen content in the hepatocyte were examined. After
16 hours of exposure, DDE and trans-nonachlor significantly decreased glucose production in the primary hepatocytes in
a concentration-dependent manner. While both DDE and trans-nonachlor decreased hepatocyte glucose production, glucose uptake and glycogen content were not significantly altered. These organochlorine effects on glucose production were
observed at non-cytotoxic concentrations of both DDE and trans-nonachlor. In summary, our current data indicate direct
exposure to select organochlorine pesticides negatively impacts glucose metabolism by decreasing hepatocyte glucose production which may promote fasting hypoglycemia.
Research Grant: Mississippi State University College of Veterinary Medicine Office of Research and Graduate Studies
Preliminary Data Grant
Student Support: The present work was supported by National Institutes of Health grant #T35OD010432
162
Beneficial effects of probiotic microbes on inflamed gut mucosa in the rhesus macaque model of HIV infection
Katti Horng, Anne Fenton, Irina Grishina, Lauren Hirao, Beau Parry, Amir Kol, Satya Dandekar
Department of Medical Microbiology & Immunology (Horng, Fenton, Grishina, Hirao, Parry, Dandekar), University
of California, Davis, Davis, California; Department of Pathology, Microbiology, and Immunology (Kol), University
of Veterinary Medicine, University of California, Davis, Davis, California
Chronic immune activation and persistence of viral reservoirs remains a challenge for achieving complete clinical resolution
in Human immunodeficiency virus (HIV) infected patients despite long-term retroviral therapy. Simian immunodeficiency
virus (SIV) infected rhesus macaques provide an excellent nonhuman primate model of AIDS that manifest clinical, immunologic, and pathologic changes similar to that of HIV infection in humans. HIV infection driven CD4+ T cell depletion
in peripheral blood is progressive over a long period of time, but very rapid and severe in gut associated lymphoid tissue
(GALT), leading to impaired mucosal immunity, gut epithelial barrier disruption, and viral persistence. Pilot studies show
that probiotic bacteria provide reversal of CD4+ T cell loss, inflammation, and metabolic disorders in HIV infection. Utilizing a ligated ileal loop model in SIV-infected rhesus macaques, we investigated chronic gut mucosal responses following
L. plantarum and B. infantis administration. Comparative analyses of viral loads, T cell subset distribution, mucosal gene
expression profiles, and epithelial integrity were performed by RT-PCR, flow cytometry, DNA microarrays, and fluorescent
immunohistochemistry. Results suggest that these commensal bacteria dampen inflammatory genes and IL-1b signaling,
which may induce cell cycling and repair epithelial integrity in gut mucosa. This study provides a unique opportunity to
characterize the molecular signatures of immune function in combination with histopathological changes in the gut epithelium during infection and following the exposure to probiotic microbes.
Research Grant: NIH R01 #AI043274
Student Support: UC Davis STAR, NIH T-35, VSTP Program
Use of the Collaborative Cross to Assay the Impact of Inter-Individual Variability on TCDD-mediated Signaling
James Hosner, Peter Dornbos, John LaPres
Department of Biochemistry and Molecular Biology, Michigan State university (James Hosner), Institute for Integrative
Toxicology, Michigan State University (Dornbros, Laprey), Center for Mitochondrial Science and Medicine, Michigan
State University, East Lansing, Michigan, USA.
Very little is known about how an individual’s genotype influences their response to chemical exposures. It is known that
there is inherent variability in this response that is based on many things, including the individual’s genotype, past exposures, age, and sex. Understanding the relationship between population-level variability and response to chemical exposures is critical for accurate risk assessment, however, inter-individual variability is not usually assessed in toxicological
studies. Addressing this knowledge gap has the potential to impact public health on many levels, so newly impregnated
females of a genetically diverse panel of mice called the collaborative cross (CC) were treated with the environmental pollutant, 2,3,7,8 tetrachlordibenzo-p-dioxin (TCDD). TCDD activates the aryl hydrocarbon receptor (AHR), a transcription
factor that regulates the expression of a wide-array of genes. Of these genes, several members of the cytochrome p450 superfamily, such has Cyp1A1 and Cyp1B1, are commonly induced upon TCDD exposure. We plan to analyze the expression
of Cyp1A1 and Cyp1B1 in the liver of CC mouse strains that were exposed to various concentrations of TCDD. Since the
CC panel is a good model for the diversity within the human population, we predict that the variation within the Cyp1A1
and Cyp1B1 expression data will give us an impact of genotype on chemical exposure response. It will allow us to address
how this variation will influence our ability to accurately assess risk. This study will add to the understanding of the role of
inter-individual variability in determining a population-level dose-response relationship along with identifying susceptible
variants associated with TCDD-induced toxicity.
Research Grant: NIH R25HL103156
Student Support: Michigan State Summer Research Program
163
Jennifer Howard, Amanda Burling, Craig Franklin
University of Missouri Veterinary Research Scholars Program (Howard) Department of Veterinary Medicine and Surgery,
College of Veterinary Medicine, University of Missouri (Burling) Department of Veterinary Pathobiology and MU
Metagenomics Center, College of Veterinary Medicine, University of Missouri (Franklin)
The gut microbiota (GM) of most species studied to date has been shown to benefit host health by aiding in digestion, immune system development and disease resistance to pathogens. Moreover, changes in GM have often been correlated with
both intestinal and extra-intestinal disease. Study of the feline GM is in its infancy and it is largely unknown how composition and/or complexity change with changes in environmental factors or disease states. Millions of domestic cats are housed
in shelters each year and these cats are exposed to many unique factors that may predispose them to disease. The aims of our
study are to 1) compare the GM of shelter housed felines to that of cats in household settings and 2) characterize the GM
of shelter felines exposed to a variety of environmental factors including, but not limited to housing style (single vs group),
perceived stress levels, and daily enrichment. To determine GM composition and diversity, DNA is extracted from fecal
samples and subjected to next generation sequencing and Qime analysis. We anticipate that significant differences in GM
composition and/or diversity will exist between shelter-housed and client-owned felines. We are also expecting to find GM
differences among felines that have differing housing settings, behaviors, fecal consistency scores, or dermal, ocular and
aural disease states. By correlating changes in specific bacterial species with environmental and disease parameters, we will
set the stage for future studies designed to establish causal roles for select bacterial species and develop novel therapeutic
measures to prevent GM shifts associated with disease.
Research Grant: University of Missouri Metagenomics Center and PI research incentive funds
Student Support: Stipend for Jennifer Howard is supported by a grant from Merial, a Sanofi Company
Teodora S. Hristova, Alexandria M. Lesicko, and Daniel A. Llano
College of Veterinary Medicine (Hristova), Neuroscience Program (Lesicko, Llano) and Department of Molecular and
Integrative Physiology (Llano), University of Illinois at Urbana-Champaign, Urbana, IL
Auditory and somatosensory structures in the brain send information to the lateral cortex of the inferior colliculus, a structure that is believed to be important in multisensory integration. Previous studies have shown that this nucleus contains a
network of modules, or compartmental zones which stain densely for certain neurochemical markers, such as glutamic acid
decarboxylase-67 (GAD-67). Though the function of these modules remains unknown, the termination patterns of sensory
inputs to the lateral cortex appear structurally similar to the distribution of neurochemical modules. The goal of the present
study is to determine whether the auditory and somatosensory inputs to the lateral cortex project into the modules, interdigitate with the modules, or show no relationship with these neurochemical structures. The auditory and somatosensory
projections to the lateral cortex were labeled with the anterograde tracer, biotinylated dextran amine (BDA) and GAD-67
immunohistochemistry was done to expose the modular network in the lateral cortex of the mouse brain. GAD-labeled
modules and BDA-labeled input terminals within the inferior colliculus were visualized using confocal microscopy, then
images were imported in Neurolucida , a software program in which axonal reconstructions were performed. Analysis revealed that somatosensory inputs from the somatosensory cortex and dorsal column nuclei project into the modules, while
auditory inputs from the auditory cortex and central nucleus of the inferior colliculus interdigitate with the modules. These
results suggest that the auditory and somatosensory inputs to the mouse lateral cortex form isolated processing pathways.
Research Grant: NIH R01 DC013073 and NSF IGERT Fellowship 0903622
164
Construction and expression of full-length canine circovirus molecular clone
Steven Hsu, Rie Watanabe, Steven Kubiski, Patricia Pesavento
Department of Pathology, Microbiology, and Immunology, School of Veterinary Medicine,
University of California - Davis, Davis, CA
Since the discovery of canine circovirus (CaCirV) in 2012, there have been multiple reports describing its association with
enteric and systemic disease in dogs and other canids. Because CaCirV can also be detected in the feces of normal, healthy
dogs, the presence of the virus alone is not useful in determining disease outcome. While continuing research on natural
cases is unequivocally valuable, the establishment of an in vitro viral culture system is necessary to identify cell targets of
infection, host-receptor interactions, viral growth rates, and factors that regulate virulence. Therefore, a full-length molecular clone of CaCirV was produced and transfected into Madin-Darby canine kidney (MDCK) and human embryonic kidney
(HEK) cell lines. Viral production and translation of viral capsid protein were analyzed using a combination of polymerase
chain reaction (PCR) and Western blotting procedure. Finally, both immunofluorescence assay (IFA) and transmission
electron microscopy (TEM) were used for localization of virus production and assembly to specific cellular compartments.
This is the first study to analyze an infectious clone of canine circovirus in an in vitro system.
Research Grant: The Morris Animal Foundation
Student Support: Student Training in Advanced Research Program (STAR)
Direct regulation of MLKL by EFhd2 to suppress the necrotic signaling pathway
Jaimie Huff, Paula J Klutho, and Christopher P. Baines
College of Veterinary Medicine, University of Missouri-Columbia, Columbia, MO
Necrotic cell death underlies several diseases such as cardiac infarction, liver cirrhosis, and renal failure. Thus researching
how necrosis is signaled is vital for advancing the treatment of multiple diseases. Necrosis occurs when TNF receptors
on cells are activated leading to the phosphorylation and activation of the kinases RIP1 and RIP3. RIP3 then binds to and
phosphorylates the pseudokinase MLKL. Phosphorylated MLKL then oligomerizes and translocates to the plasma membrane where it forms a non-specific pore that kills the cell. We have previously identified EFhd2 as a novel MLKL-binding
protein and shown that it can suppress TNF-induced necrosis. However, the molecular mechanism by which EFhd2 inhibits
necrosis remains to be elucidated. We hypothesized that EFhd2 inhibits necrosis by preventing the phosphorylation, and
hence oligomerization, of MLKL. To test this, we are overexpressing (adenovirus) or depleting (siRNA) EFhd2 in mouse
embryonic fibroblasts (MEFs) and examining the effects on TNFa-induced necrosis using Sytox staining. Western blots for
the phosphorylated forms of RIP1, RIP3, and MLKL are being run to assess activation of the necrosis pathway. Finally,
we are determining the effects of EFhd2 manipulation on MLKL oligomerization using non-reducing westerns. We expect
to see decreased cell death in the EFhd2 overexpressing MEFs and more cell death in the EFhd2-depleted MEFs. We also
expect to see reduced phosphorylated and oligomerized MLKL with the MEFs overexpressing EFHD2 and the opposite in
the MEFs with reduced EFhd2. Such results would indicate that EFHD2 inhibits necrosis through the prevention of MLKL
activation, information that can be used to develop new treatments for several diseases.
Research Grant: Research Grant: NIH R01 HL094404
Student Support: Student Support: IDEXX-BioResearch
165
LPS stimulated GRK2 expression in response to novel anti-inflammatory drugs in raw 264.7 macrophage cells
Courtney D. Hughes, Michael Steury, Narayan Parameswaran
Department of Physiology, Michigan State University, East Lansing, MI-48823
G-protein coupled receptor kinases (GRKs) have been known to play a crucial role in the pathogenesis of inflammatory diseases. GRKs are responsible for regulating desensitization of G-protein coupled receptors (GCPRs), the largest and most diverse class of membrane receptors in eukaryotes. Specifically, GRK2 activity and expression levels have been widely shown
to be effected by inflammatory disease conditions. Previous studies have shown that through the stimulation of murine
peritoneal macrophages with LPS, GRK2 expression levels will increase. Therefore, an expansion on this data will be done
by stimulating raw 264.7 cells (a murine macrophage cell line) with LPS to observe the cellular GRK2 expression. Furthermore, novel anti-inflammatory drugs, LG100268 and I-BET, will be tested with LPS stimulated macrophage cells to test the
effect these drugs have on GRK2 expression. Raw 264.7 macrophage cells were grown and stimulated with LPS (1ug/ml)
and treated with novel anti-inflammatory drugs and to determine their effect on GRK2 expression, GRK2 was measured at
both 6 and 24 hours. GRK2 expression levels were determined through SDS-PAGE electrophoresis and quantified through
densitometry. This study will expand on the relationship between GRK2 expression and its response to LPS specifically in
the context of the anti-inflammatory drugs LG100268 and I-BET. Knowledge on GRK2 mechanisms and implications on
inflammatory diseases and responses will allow for future research to be done on novel anti-inflammatory drugs to ensure
anti-inflammation and increase effectiveness in new market drugs.
Generation of functional b islet cells from feline adipose-derived multipotent stromal cells
Peyton Hughes, Wei Duan, and Mandi J. Lopez
Laboratory for Equine and Comparative Orthopedic Research, Department of Veterinary Clinical Sciences, School of
Veterinary Medicine, Louisiana State University, Baton Rouge, LA
Adipose-derived multipotent stromal cell (ASC) therapies are becoming increasingly popular. Pancreatic b islet cells from
multipotent stromal cells have been tested to treat diabetes in mice and humans. Diabetes mellitus is a serious feline problem that is increasing in prevalence. We hypothesized that feline ASCs can differentiate into functional b islet cells (BIC).
ASCs were isolated from reproductive adipose tissue routinely removed during feline sterilization and grown to passage (P)
1. Cell doubling times (CD) and colony forming unit (CFU) frequencies were used to determine expansion rate and multipotentiality of the cells. Passage 2 cells were cultured in BIC induction medium on Ultra Low Attachment cultureware for
18 days. Cells were then evaluated with Dithizone (DTZ) staining for high concentrations of zinc associated with insulin
storage in pancreatic b cells and immunohistochemistry for insulin. Female and male cells doubled in 0.82 6 0.04 and 0.93
6 0.04 days (mean 6 SEM), respectively. There was no difference in adipocytic, osteoblastic or fibroblastic CFU frequencies or CD between sexes. Positive DTZ and insulin staining in cells cultured in BIC differentiation medium confirmed
successful BIC differentiation. Feline ASCs were able to expand in vitro and exhibit multipotency. With no significant
differences between sexes, both male and female reproductive tissues can be used to isolate feline ASCs. The viability of
the cells after islet induction and presence of insulin confirm that feline ASCs can differentiate into insulin-producing BICs.
These results indicate that implantation of BICs from stem cells may provide a new therapeutic option to treat or cure feline
diabetes.
Research Grant: Winn Feline Foundation
Student Support: LSU School of Veterinary Medicine, Laboratory for Equine and Comparative Orthopedic Research
166
Janna Hunt, Allison J. Richard, Hardy Hang, Carrie Elks and Jacqueline M. Stephens
LSU School of Veterinary Medicine, Baton Rouge, LA. Pennington Biomedical Research Center, Baton Rouge, LA.
Significant advances towards understanding obesity and diabetes have been made by studying the function of proteins that
modulate fat cell function. Our studies focus on understanding the function of STAT5 proteins in fat cells to determine how
they contribute to lipid metabolism and insulin action. We have generated mice that lack both STAT5 genes specifically in
adipocytes. Our specific aim is to assess the contribution of adipocyte STAT5 proteins to glucose and lipid metabolism in
adipocytes and overall energy balance. We will complete phenotypic analysis of the male mice and then assess the lipid
responses and gene expressions for both male and female mice. We hypothesize that adipocyte-specific knockout of both
STAT5 genes in vivo will affect fat cell functions and whole body glucose disposal and reveal the primary function(s) of
STAT5 proteins in adipocytes.
Research Grant: R01DK052968-15 NIH to JMS
Student Support: NIH
Jacquelyn A Hunter, James Weger, Greg Ebel, Mark Stenglein.
Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences,
Colorado State University, Fort Collins, CO
Pathogens vectored by ticks pose a significant danger to human and animal health. The other viruses and microorganisms
that infect ticks may alter their ability to harbor and transmit these pathogens. Additionally, viruses and other microbes
that are at present specific to ticks have the potential to become human or animal pathogens. Despite the risk, the tick virome and microbiome remain largely unexplored. We therefore used metagenomic sequencing to comprehensively characterize the microbial communities associated with wild ticks. For a pilot study, we prepared sequencing libraries from RNA
extracted from 16 Ixodes scapularis ticks that were collected in northern Wisconsin. We performed paired-end Illumina
sequencing and taxonomically assessed non-tick sequences. We identified a number of organisms, including human and
animal pathogens. For instance, in 13/16 ticks, we identified Borrelia burgdorferi, the causative agent of Lyme disease.
We also identified the sequences of various virus in 16/16 ticks, including sequences from South Bay Virus, Blacklegged
tick virus 1 & 2, and Suffolk virus. Expanded studies will begin to investigate if and how levels of these viruses or other
microbes correlate with the presence of B. burgdorferi and other pathogens.
Research Grant: Colorado State University College of Veterinary Medicine and Biomedical Sciences, and the Department
of Microbiology, Immunology, and Pathology
167
Degradation of complement in the presence of Schistosoma mansoni in human serum
Melissa Icaza, Patrick Skelly, Akram Da’darah
Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine, Tufts University,
North Grafton, MA
Schistosomiasis is a parasitic disease of major public health concern, infecting approximately 200 million people and countless animals every year. Despite living in the blood stream of immunocompetent hosts, they are largely refractory to the host
immune system, including the complement cascade. Previous work in this lab has indicated that complement protein C3 is
degraded more rapidly in the presence of the worms and has suggested C4, Factor B, and Clusterin may be similarly affected. In this project, we demonstrate that C4 and Factor B are cleaved more rapidly in the presence of the schistosomes. In
contrast, no effect was seen on the degradation of Clusterin. Immunofluorescence assays using live schistosomes show that
C4 and Factor B both attach to the surface of schistosomes. This suggests that the worms are interacting with complement
components yet do not appear to be killed by complement. The enhanced degradation of selected complement components,
as demonstrated here, represents a potential pathway for schistosomes to circumvent this aspect of host immunity.
Research Grant: NIH
Student Support: In part supported by a grant from NIH (5T35OD010963-07)
Lauren Ienello, Michelle Carstensen, Larissa Minicucci, Erik Hildebrand, Chris Jennelle, Glenn DelGuidice,
Jed Overmann.
Minnesota Department of Natural Resources, University of Minnesota College of Veterinary Medicine
Free-ranging moose (Alces alces) in Minnesota have recently been declining and health-related causes comprise a majority
of documented deaths in the latest studies. The use of blood profiles to assess the overall health of animals is important in
understanding influences on survival. Additionally, reference intervals for hematological and serum biochemistry parameters are lacking for wild moose populations. The Minnesota Department of Natural Resources has compiled hematological
and serum biochemistry profiles from presumed healthy hunter-harvested (n=223, 99% males, 1% females, 2010-2012) and
live-captured (n=331, 25% males, 75% females, 2010-2015) moose. Live-capture blood samples prove to be vital in the
assessment of wild populations, and hunter-harvested samples may provide additional value. Presented are the established
normal reference intervals and a preliminary investigation of these blood profiles to improve our understanding of what
constitutes good health in free-ranging moose. Reference limits were established following the ASVCP Quality Assurance
and Laboratory Standards Committee (QALS) Guidelines for the determination of Reference Intervals in Veterinary Species
and other related topics: SCOPE. Biological factors examined include age (0.5-16 years), sex, body condition, time of year
and fate. Pre-analytical factors examined include blood handling times from time-of-kill or capture to time-of-blood spun.
An in depth literature review has also been conducted to compare these findings from free ranging-moose in Minnesota to
other wild and captive cervid populations. To our knowledge, this is the largest blood profile dataset of free-ranging moose
and the first to include hunter-harvested data.
Research Grant: Minnesota Department of Natural Resources.
Student Support: Merial Limited and the College of Veterinary Medicine, University of Minnesota.
168
Do honey bee colonies (Apis mellifera) prefer feed contaminated with neonicotinoids?
Claire Janse van Rensburg, Sarah Wood, Ivanna Kozii, Roman Koziy, and Elemir Simko
Department of Veterinary Pathology, Western College of Veterinary Medicine, University of Saskatchewan,
Saskatoon, SK
Neonicotinoids are the most commonly used insecticides worldwide. High mortality rates in honey bee colonies have drawn
attention to widespread neonicotinoid use. Laboratory studies confirm that sublethal neonicotinoid exposure negatively
affects honey bee health, however more research is required to study the impact of neonicotinoids on whole colonies under
natural conditions. Kessler et al. reported that small groups of bees prefer feed containing neonicotinoids under laboratory
conditions (Nature, 2015, 531:74-88). The objective of this study was to investigate whether complete honey bee colonies
with preserved eusocial order prefer feed containing neonicotinoids. Top-feeders were used to give colonies free choice of
sucrose syrup containing clothianidin, imidacloprid, or thiamethoxam at 0nM, 4nM, 40nM, and 400nM (n=4 hives/neonicotinoid); bees had direct access to syrup from within the hive. Changes in the weights of feeders were used as a proxy to
measure syrup consumption. Each concentration was expressed as a percentage of the total consumed feed in three different
feeding trials and analyzed by ANOVA. Results indicated that honey bee colonies do not prefer feed containing higher concentrations of neonicotinoids. Syrup consumption ranged from 24.3-25.8%, 24.5-25.5%, and 23.6-26.2% between the four
concentrations of clothianidin, imidacloprid, and thiamethoxam respectively. The neonicotinoid concentration in the syrup
did not influence its consumption. These results contrast with studies performed under artificial conditions and suggest that
honey bees with preserved eusocial order at the colony level are not attracted to contaminated crops. This offers encouragement to beekeepers and agriculture alike.
Research Grant: Saskatchewan Beekeepers Association, Western Grains Research Foundation, SaskCanola, Canadian
Honey Council, Pollinator Partnership, NAPPC, WCVM Wildlife Health Fund
Characterization of the skin microbiome of dogs with strong body odor and the effect of a spot-on product
Roxanna Jeffreys, Courtney M. Smith, Brandon Dominguez, Adam P. Patterson, Sara Lawhon,
and Aline Rodrigues Hoffmann
Department of Veterinary Pathobiology (Jeffreys, Smith, Lawhon, and Rodrigues Hoffmann), Department of Large
Animal Clinical Sciences (Dominguez), and Department of Small Animal Clinical Sciences (Patterson),
Texas A&M University College of Veterinary Medicine and Biomedical Sciences, College Station, TX
Next-generation sequencing (NGS) analysis has been used to evaluate the microbiota on canine skin. The aim of this study
was to investigate bacterial and fungal microbiota on the skin of dogs with strong body odor and to determine if body odor
was mitigated with the use of Dermoscent Essential 6 Spot-On Product. We hypothesized that dogs with strong body odor
would have a dysbiosis of their skin microbiota with an increased abundance of Staphylococcus and Corynebacterium sp.
To test this hypothesis, we collected surface skin swabs from two body regions (axilla and dorsum) from dogs with or without strong body odor at day 0 and after 4 weeks. Dogs with strong body odor received either a topical essential oil product
treatment or a placebo vehicle once per week for four weeks. DNA was extracted and sent off for 16S NGS. Samples were
also evaluated using quantitative PCR for Malassezia, Corynebacterium, Staphylococcus, and Staphylococcus pseudintermedius. At day, 0 the strong odor body group had a significantly higher average body odor grading compared to the control
group. After four weeks of treatment with Dermoscent Essential 6 Spot-On Product, the average body odor grade was
significantly reduced for the treatment group, but not for the placebo group. NGS and quantitative PCR are underway. The
information obtained in this study will be used for identification of microorganisms responsible for odor in dogs, and can be
further used to support the development and improvement of topical veterinary products used to diminish strong body odor
and maintain the protective skin barrier.
Research Grant: Laboratoire de Dermo-Cosmetique Animale
169
Impact of Helicobacter pylori on the infant lung: airway microbiome and immunomodulation of airway epithelium
Monica T. Jimenez, Clarissa S. Rocha, Matthew Rolston, Lori M. Hansen, Jay V. Solnick, and Lisa A. Miller
California National Primate Research Center (Jimenez, Miller), School of Medicine (Rocha, Rolston, Solnick),
and Center for Comparative Medicine (Solnick, Hansen), University of California, Davis, Davis, CA
An inverse relationship between Helicobacter pylori infection and childhood asthma has been demonstrated by epidemiologic studies, however the mechanism is unknown. We hypothesized that H. pylori infection may alter the microbiome of the
lung during infancy. We further speculated that aspiration of H. pylori from the gastrointestinal tract could directly induce
innate immune responses in airway epithelium. To test this hypothesis, we utilized the infant rhesus macaque as a model of
postnatal development. Gastric biopsies, BAL, and oral swabs were collected from 25 monkeys at 4-7 months of age for 16s
sequencing. The effect of H. pylori on airway epithelium was determined by cultures with the 16-HBE cell line and rhesus
primary tracheobronchial epithelial (TBE) cells. Contributions of the H. pylori cag pathogenicity island (cag PAI) were
measured using cag knockouts of H. pylori strains PMSS1 and J166. 16s sequencing data showed that the lung and oral
microbiomes of infant rhesus macaques have distinct microbial populations. The oral cavity is dominated by Proteobacteria
and Firmicutes while the lung is dominated by Actinobacteria. Following H. pylori infection in vitro, IL-8/IL-6 secretion
by airway cultures was significantly increased in a MOI- and age-dependent manner. H. pylori cagY and cagA knockouts
reduced IL-8 secretion in 16-HBE and TBE cultures. Our data supports the idea that the lung microbiome is not an amalgam
of oral aspirates; rather it represents a distinct microbiome. H. pylori can directly promote airway epithelial cell expression
of proinflammatory cytokines in an age- and cag PAI-dependent manner, suggesting a potential window of susceptibility for
immunomodulation of the lung during early life.
Research Grant: NIH R56AI091893 and P51 OD011107
Student Support: NIH T32 Training Grants 5T32GM007170-41 and 4T32AI070077-09
MARCKS protein as a therapeutic target in the treatment of equine asthma
Cathy Johnson, Mary Sheats
Department of Clinical Sciences, North Carolina State University CVM, Raleigh, NC
Equine asthma is a debilitating and costly disease of horses that negatively impacts their performance and overall well-being. Exposure to environmental allergens elicits an overwhelming inflammatory response in asthmatic horses, leading to
the accumulation of neutrophils in the airways and subsequent injury to respiratory tissues. Alternative therapies are needed
that specifically target inhibition of this damaging neutrophil response. One key regulator of neutrophil functions previously
identified by our lab is the MARCKS (Myristoylated alanine-rich C kinase) protein. Given the prominent role that neutrophils play in equine asthma, we hypothesize that MARCKS is integral to the inflammatory response within the airways of
asthmatic horses and that inhibition of MARCKS will reduce this response. We will first determine whether MARCKS
expression is up-regulated in the pulmonary cells of asthmatic horses. The quantity of MARCKS protein in pulmonary
lysates from 6 healthy and 6 asthmatic horses will be evaluated ex vivo using biochemical analysis. Antibody selection is
currently being optimized for this experiment. Next, we will evaluate whether MARCKS inhibition, via the MANS peptide,
will attenuate the ex vivo cytokine response of LPS-stimulated alveolar macrophages from asthmatic horses. Cytokine expression and release will be measured via qRT-PCR and ELISA, respectively. This experiment is currently being conducted
in LPS-stimulated equine peripheral blood leukocytes for future application in equine alveolar macrophages. Overall, our
goal is to identify novel treatment options for horses with asthma and successful completion of this project will establish
MARCKS as a viable target for such therapies.
Student Support: Fund for Discovery, Merial Veterinary Scholars Program
170
Susceptibility of equine monocyte subsets to equine herpes virus type I infection
Lauren A. Johnson, Priscila B. S. Serpa, Tracy Stokol
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University,
Ithaca, NY
Equine herpes virus type I (EHV-1) is an endemic, highly contagious equine pathogen that causes respiratory illness, abortion, and encephalomyelopathy. Outbreaks can cause high morbidity and significant financial loss, particularly in racing
and breeding operations. While vaccines are available, they have limited effectiveness and do not protect against all virus
strains. To inform future vaccine and treatment development, a greater understanding of the interactions between EHV-1 and
immune cells, such as monocytes, is crucial. Pro- and anti-inflammatory monocyte subsets have been identified in the blood
of other animals and humans. Viruses such as HIV differentially infect and alter the proportion of these monocyte subsets,
which may reduce the efficacy of the antiviral immune response. EHV-1 can infect equine monocytes and virus DNA can be
amplified from monocytes isolated from the blood of infected horses. A subset of equine monocytes express the scavenger
receptor CD163, a marker of anti-inflammatory monocytes. We hypothesized that equine CD163+ and CD163- cells will
show different susceptibility to EHV-1. To test this hypothesis, we are infecting peripheral blood mononuclear cells with
EHV-1 strains that express green fluorescent protein and utilizing antibody staining and flow cytometry to identify and
quantify the percentage of infected CD163+ and CD163- monocytes. Additionally, we are purifying and separating both
monocyte subsets from equine blood and examining their susceptibility to infection. The second approach will allow future
investigation of cytokine production by each subset in response to EHV-1 infection.
Research Grant: Morris Animal Foundation D16EQ-815
Student Support: Office of Graduate Education- Cornell University College of Veterinary Medicine
Coagulopathy in Crotalus viridis envenomation, attenuation by carbon monoxide releasing molecule - 2 in vitro
Tyler Ellis Johnson, Amy Bell, Raegan J. Wells, Christine S. Olver
Department of Microbiology, Immunology and Pathology (Johnson, Olver), College of Veterinary Medicine
and Biomedical Sciences, Colorado State University, Fort Collins, CO; Veterinary Specialty Hospital (Bell),
San Diego, CA; BluePearl Veterinary Partners Arizona (Wells), Phoenix, AZ
Rattlesnake venom fibrinogenases cause decreased clot strength, and in the presence of tissue plasminogen activator (tPA)
in vitro, a markedly increased rate of clot lysis. Carbon Monoxide Releasing Molecule -2 (CORM-2), an emerging therapeutic in human medicine, enhances plasmatic coagulation and attenuates fibrinolysis in vitro in human, rabbit, and horse
plasma, and ameliorates hypocoagulation and hyperfibrinolysis secondary to venom exposure in human plasma in vitro.
Once solubilized, CORM-2 liberates carbon monoxide (CO). CO interacts with a heme group on fibrinogen, changing
its configuration so that the fibrin clot is strengthened and more resistant to fibrinolysis. We hypothesized that CORM-2
enhances coagulation and attenuates fibrinolysis in canine plasma exposed to C. viridis venom. We measured the effects
of C. viridis venom on clot strength, rates of coagulation and fibrinolysis in both pooled canine plasma and plasma from
individual naturally envenomated dogs, with and without CORM-2, using thromboelastography (TEG). We tested venom
effects on coagulation using tissue factor (TF) activated TEG, and on both coagulation and fibrinolysis using TF activated
TEG with added tPA. We found that 17.9mg/mL of venom causes a mean 26.4% decrease in clot strength, a 61.8% decrease
in maximum rate of thrombus generation, 75% faster clot lysis, a 226% increase in maximum rate of lysis , and a 92% decrease in total clot life span (CLS). CORM-2 ameliorated these effects, increasing CLS in the presence of venom by 603%.
Additionally, we showed that CORM-2 has identical effects in vitro on plasma from naturally envenomated dogs. Future
studies with CORM-2 and venom in whole blood and with other snakes’ species are to come.
Research Grant: The Center for Companion Animal Studies, Young Investigator Grant; Colorado State University
Student Support: Merial, Ltd
171
The Golden Retriever Lifetime Study: factors affecting owner compliance after the first year of enrollment
Melissa Jones, Audrey Ruple, Missy Simpson, and Rodney Page
Departments of Veterinary Clinical Sciences and Comparative Pathobiology, College of Veterinary Medicine,
Purdue University, West Lafayette, IN (Jones, Ruple); Morris Animal Foundation (Simpson); Flint Animal Cancer Center,
College of Veterinary Medicine and Biomedical Sciences, Colorado State University, Fort Collins, CO (Page)
Longitudinal cohort studies are a valuable way to obtain a large amount of information about a population over a period of
time. In veterinary medicine, few large cohort studies have been undertaken due primarily to the great amount of time and
expense required to complete them. The Golden Retriever Lifetime Study (GRLS) currently being conducted by Morris
Animal Foundation is the largest dog cohort study of its kind ever initiated in the U.S., and understanding factors that affect
owner compliance with research requirements will help improve cohort study design and participant recruitment in the future. Thus, the objective of this study was to determine factors affecting dog owner non-compliance by the end of the first
year of GRLS based on information provided by owners at the time of enrollment. The study population consisted of Golden
Retrievers (n=3044) whose owners elected to participate in GRLS. Logistic regression modeling was used to determine
factors associated with owner non-compliance at the end of the first year post-enrollment. The results showed that owners
of dogs that sleep in areas other than their bedroom are at increased risk for non-compliance, especially those with dogs that
sleep in a garage at night. Owners who do not bathe and/or groom their dogs at home and who do not vaccinate their dogs are
also at increased risk for non-compliance. This study shows that survey questions about a dog’s sleeping location at night,
home grooming habits, and vaccination status are important indicators of an owner’s likelihood of compliance in a cohort
study. Similar questions could be used in future cohort studies to screen for owners likely to be compliant, thus increasing
the likelihood of success of the study.
Ruby E. Jong, Simona Kraberger, Justin S. Lee, Esther Musselman, Sue VandeWoude
Department of Microbiology, Immunology, and Pathology, College of Veterinary Medicine and Biomedical Sciences,
Feline Coronavirus (FCoV) is an enveloped RNA virus (~29kbp) that infects domestic cats. FCoV is highly prevalent,
detected in approximately 90% of multi-cat households and 50% of single-cat households. In 5-10% of infections, FCoV
undergoes genetic changes that can lead to a syndrome known as Feline Infectious Peritonitis (FIP). FIP causes a fatal
systemic disease and is a leading cause of death in young cats less than 3 years of age. The aim of this study was to survey
the prevalence of FCoV in a specific pathogen-free (SPF) cat colony (n=38) with a history of 50% FCoV seroprevalence.
The goal was to develop and apply an optimized PCR protocol on blood, saliva and fecal samples, and compare results
with FCoV ELISA serology diagnostics. RNA was isolated from peripheral blood mononuclear cells (PBMC), plasma and
feces, and subjected to a validated and sensitive nested PCR assay. Preliminary PCR screening results indicate 40% of cats
tested positive for FCoV in at least one sample type. Sample types from each cat were not consistently positive. Eight (21%)
plasma samples and nine (24%) PBMC samples were PCR positive. Four (57%) of seven fecal samples tested to date were
positive. Based on these preliminary results, feces seem to be the most reliable source for FCoV screening. Sporadic results
from individual cats suggest PBMC and plasma samples are not reliable sample types, but do indicate low level viremia in
a significant number of asymptomatic cats. Future studies will include screening of the remaining fecal samples, saliva samples, and comparing PCR to serologic tests. We will also optimize a qPCR protocol to investigate viral load and elucidate
FCoV transmission and infection characteristics in a closed colony.
Student Support: Merial, Ltd
172
Do you see what I see? Inter-rater reliability of a canine welfare assessment used in dog breeding kennels
Mary E. Jordan, Amy Bauer, and Candace Croney
Purdue University CVM, West Lafayette, IN (Jordan), Center for Animal Welfare Science and Department
of Comparative Pathology, Purdue University, West Lafayette, IN (Croney, Bauer)
Accurate assessments of dog behavior and welfare are important to determine the wellness of kenneled dogs and to evaluate the adequacy of their environments. These evaluations are especially important in facilities where dogs are housed for
extended periods of time such as breeding kennels or shelters. In order for the results of canine welfare assessments to be
valuable, they must be proven valid and reliable. In this study, a canine welfare assessment was developed to evaluate the
overall wellness of breeding dogs at three Amish breeding facilities in southwest Indiana. Two novice raters evaluated the
behavior and welfare of 20 dogs at each breeding facility in order to assess the inter-rater reliability (IRR) of the tool. IRR
was calculated by using Cohen’s Kappa in SPSS. When all facilities were included in the analysis, raters gave the same
score 81.13% of the time. When chance agreement was accounted for by use of kappa, there was moderate agreement, kappa= 0.626, p<0.005. Agreement improved when dogs were evaluated with their primary caretakers present as raters gave
the same score 85.71% of the time and when chance agreement was accounted for there was moderate agreement, kappa=
0.728, p<0.005. Because there was moderate agreement, training of raters is recommended to further improve reliability
results. Altogether, this canine welfare assessment is a useful tool for evaluating the wellness of kenneled dogs in breeding
facilities and may be helpful in developing corresponding recommendations for individual dogs.
Research Grant: World Pet Association, Pet Food Institute
Student Support: Merial, Purdue University College of Veterinary Medicine
The efficacy of counter conditioning to attenuate signs of fear in dogs at the veterinary clinic.
Jalika Joyner, Sherrie Yuschak, Katherine Walker, John Majikes, Justin Kuhn, Rita Brugarolas, Alper Bozkurt,
David Roberts, and Barbara Sherman
Department of Clinical Science- College of Veterinary Medicine (Joyner, Yuschak, Sherman), Department of Electrical
and Computer Engineering (Walker, Kuhn, Brugarolas, Bozkurt), Department of Computer Science (Majikes, Roberts),
North Carolina State, Raleigh, NC
Fearful dogs at the veterinary clinic pose healthcare, welfare, and safety concerns. Our goals are to characterize the behavioral and physiological traits of fearful dogs and to determine if fearful dogs treated with on-site counter conditioning,
a behavior modification technique, show reduced signs of fear compared to untreated fearful dogs. Healthy client-owned
dogs with a history of fear in the veterinary clinic were enrolled and assigned a priori to treatment or control groups. Prior
to the start of the session, each dog was instrumented with non-invasive heart and respiratory rate recording devices and
aural body temperature was taken. The owner was seated with his/her dog in a veterinary consultation room for 21 minutes.
During this time, the treatment group received counter conditioning instructions from a veterinary technician, while the
control group did not. We video recorded each dog’s behavior and recorded its physiological measures over seven 3-minute
time intervals. After the session, the post-session aural body temperature was collected. A multivariate statistical model will
be used for data analysis. This study is currently on-going. We hypothesize that the behavior signs of fear will be correlated
with physiological signs of stress. Further, we predict that dogs exposed to counter conditioning will exhibit a greater reduction in behavioral signs of fear, heart rate, respiratory rate, and body temperature over time compared to control dogs. The
null hypothesis is that counter conditioning does not significantly decrease signs of fear when compared to controls. This
study will enhance our understanding of how to attenuate fear in canine veterinary patients.
Research Grant: The National Science Foundation #557751
Student Support: The National Science Foundation #557751 Fund for Discovery NCS-College of Veterinary Medicine
173
The effect of chronic Bisphenol A exposure on estrogen-sensitive gene expression (ER a & ER b, ERR g)
Parrish D. Kelley-Collier, Robert Crawford, Norbert E. Kaminski
Department of Pharmacology and Toxicology (Crawford, Kaminski), Comparative Medicine and Integrative Biology
program (Kaminski), Center for Integrative Toxicology (Crawford,Kaminski), Michigan State University, East Lansing ,
MI 48824.
Abstract: Production of Bisphenol A (BPA) worldwide is enormous, over 6 billion pounds a year worldwide. It uses include
binding with polycarbonate plastics, baby bottles, water bottles, medical devices, compact discs, dental sealants and canned
processed foods. BPA has been shown to possess estrogenic activity, because of this it is in a drug class called an Endocrine
Disrupting Chemical (EDC). These endocrine disruptors can either mimic endogenous hormones, causing overstimulation,
or binding to a receptor and block the endogenous hormone from binding. Numerous studies show that BPA binds estrogen
receptors alpha and beta (ER a, ER b), estrogen related receptors gamma (ERR g). However, it is poorly understood if the
levels of these receptors change in leukocytes as a direct result in BPA signaling, especially with ERR g. Conformational
changes to constant Bisphenol A low dosage exposure to these receptors can cause an increase or decrease in leukocyte
expression of these receptors ether via receptor desensitization, or through increased degradation which could impair the
ability of leukocytes to respond to endogenous estrogen. This study will determine if BPA chronic exposure changes the
mRNA expression levels of ERa and ERb and ERRg in rat splenocytes at post natal day 21 that have been dose with BPA
(2.5 and 25000 ug/kg/day). The hypothesis of this study is that exposure to chronic exposure to BPA will decrease estrogen
receptor expression (ER Alpha & Beta, ERR gamma).
Research Grant: National Institute of Health (R25HL103156)
Student Support: Student Support: SROP program (MSU), Institute of Integrative Toxicology.
Using in-lab evolution to explain how protozoan parasite Toxoplasma gondii is such an exceptional generalist
Chantelle M. Khambholja, John C. Boothroyd, Adit Naor, and Terence C. Theisen
College of Veterinary Medicine, Washington State University, Pullman, WA 99164 (Khambholja) Department
of Comparative Medicine, Stanford School of Medicine, Stanford, CA 94305 (Khambholja), and Department
of Microbiology and Immunology, Stanford School of Medicine, Stanford, CA 94305 (Boothroyd, Naor, Theisen)
Toxoplasma gondii is one of the world’s most accomplished parasites. With members of the family Felidae serving as its
definitive host, the obligate intracellular protozoan parasite Toxoplasma possesses the astounding ability to infect virtually
all endothermic vertebrates and all nucleated cell types. Possessing a worldwide distribution, it is one of the most ubiquitous
known parasitic infections in existence, infecting 30% of the human population. The question of what enables Toxoplasma
to infect such an array of hosts is the subject of this research. Previous research has demonstrated that the SAG1-related
sequence (SRS) family of surface antigen genes potentially serve at least two roles as immunomodulators and ligands to
enable adhesion. Thus, post-infection alterations in this family in different host species could enable rapid parasitic adaptation and successful invasion. RNASeq data demonstrated variations in transcript abundance following passaging through
various intermediate species’ cell lines. The data additionally indicated mRNA transcript level alterations of several SRS
cell surface proteins, with up to 30-fold increases in certain species. Therefore, we hypothesize that Toxoplasma alters its
surface proteins to suit individual hosts to invade and persist. The questions remain of how fast this occurs and to what degree. Through observing short term changes in transcript levels, we can shed light on this aspect of parasite biology. qPCR
assays in conjunction with a time-course experiment and in-lab evolution study allow a more precise understanding of said
time-scale. By understanding how Toxoplasma successfully invades we can work towards preventing infection in vulnerable
populations.
174
Michael Z. Khan, Kevin J. Kroll, Nancy D. Denslow
College of Veterinary Medicine (Khan) Department of Physiological Sciences (Kroll, Denslow)
Corexit 9527© is a commonly used dispersant in the process of cleaning up oil spills. Dispersants solubilize the hydrophobic oil, turning the large slicks into small micelles. This sinks the oil into the water and possibly makes it bioavailable
for bacterial degradation. However, the dispersants also make the toxic and mutagenic polycyclic aromatic hydrocarbons
(PAHs) more available for aquatic organisms, and can have a profound impact on the native species’ mortality and the
ecosystem’s sustainability. Menidia beryllina (Atlantic Silverside) is a common fish that resides along the Atlantic and Gulf
coast, site of the Horizon oil spill. Pervious work with these animals has shown exposure to oil and dispersants results in
reduced cardiac output. In this study we look at the bioenergetics of the Silverside embryo (48 hours post-fertilization) to
determine if exposure to the water accommodated fraction of the oil, dispersant, and the active ingredient butoxyethanol
impair basal respiration, maximal respiratory capacity and respiratory reserve. We pair this information with cardiac and
mitochondrial gene expression to demonstrate the chemicals are acting on a molecular level, damaging the cardiac myocyte’s ability to properly respire. If mitochondrial respiration is inhibited there will be a significant decrease in the amount
of ATP available for the embryo inhibiting its growth and development, ultimately leading to increased mortality and severe
decrease of the native population.
Research Grant: CVM Faculty Research Development Program
Accuracy of PetPace Collar Pulse Rate Compared to Holter Monitor Heart Rate in Dogs
Eunbee Kim and Robert Sanders
Michigan State University CVM, East Lansing, MI
One of the most common supraventricular tachycardias in dogs is Atrial Fibrillation (AF) but its non-specific clinical signs
make it is difficult to diagnose. However, it can be done definitively with electrocardiography. ECG is useful during the
treatment and management of AF. An in-house ECG is not ideal because of its time constraints and possibility of a higher
heart rate due to stress. The gold standard for at-home ECG monitoring is a Holter monitor due to its accuracy and low
level of invasiveness. Emerging technology has allowed for the development of collars such as PetPace that are even less
invasive, more cost effective, and claim to effectively monitor a pulse rate. If this collar can accurately measure pulse rate in
dog, this could be advantageous in the future for the diagnosis and treatment of AF. To test the baseline accuracy of the pulse
rate measured by the new device, 12 dogs wore the PetPace collar and a Holter monitor simultaneously for a 24-hour period
to compare the averages collected by both methods during a 30-minute period of rest, activity, a walk. The Holter monitor
was consistently able to provide a 2-minute averages for the entire 24 hours while the PetPace collar was significantly less
successful. When looking at the analogous data between the Holter monitor and collar during 3 30 minute periods, there was
no significant difference between the two. The type of activity predicted the ability of the collar to obtain a pulse rate with a
rest state being most likely and a walk state being least likely. The collar’s 2 minute pulse averages were within 10 beats of
the Holter’s averages 65.8% of the time. These findings provide insight into the clinical applicability of the PetPace collar.
Research Grant: MSU CVM Department of Small Animal Clinical Science Research
Student Support: Merial/Michigan State University College of Veterinary Medicine and Graduate School Fellowship
175
Isolation and cloning of feline Macrophage Colony Stimulating Factor for optimizing macrophage differentiation
Sohyun Kim and Yvonne Drechsler
College of Veterinary Medicine, Western University of Health Sciences, Pomona, CA
Macrophage Colony Stimulating Factor (CSF-1) is an essential cytokine for macrophage growth and survival, required for
stimulating differentiation and proliferation of peripheral blood monocytes into macrophages. In human and mouse models,
macrophage cell culture techniques have been optimized by using commercial CSF-1 to improve cell survival and differentiation in research; but this reagent is not available for felids. Thus, the main goal of this project is to clone and express the
feline CSF-1. I used screening primers made from predicted feline CSF-1 sequence (NCBI GenBank) to run RT- PCR on
feline blood samples and cell lines. This initial set of primers did not amplify products of the expected band sizes despite
various attempts. To determine the cause of failure, I used Clustal Omega to perform a comparative study of the canine,
feline, and tiger predicted CSF-1 sequences from two sources (NCBI GenBank and Ensembl) to define the gene structure.
This study revealed that there was a highly conserved exon among the three species, so I constructed a new screening primer
to target that specific portion of the gene. There was also a discrepancy in the start codon of the feline CSF-1 gene between
the two sources. The Ensembl-derived annotation start codon is predicted to have a Kozac’s sequence adjacent to the start
codon, and is the basis for our new amplification strategy.
Research Grant: Western University Summer Research Fellowship Program
Student Support: Western University Summer Research Fellowship Program and CVM Veterinary Scholars Program
Ashley J. Kirby, Amanda N. Kortum, and Jeffrey A. Yoder
Department of Molecular and Biomedical Sciences and Comparative Medicine Institute, College of Veterinary Medicine,
North Carolina State University, Raleigh, NC.
Tripartite motif containing (TRIM) proteins, which mediate the ubiquitination of a wide range of substrates, are highly conserved across vertebrate lineages. TRIM9 is highly expressed in neurons where it plays a role in regulating axonal migration.
Our lab recently has shown that Trim9 is important for macrophage motility in zebrafish (Danio rerio). In contrast, very
little is known about the function of TRIM67, which is highly similar to TRIM9. In one study, macrophage dendritic-like
cells of Atlantic salmon (Salmo salar) exposed to infectious salmon anemia virus showed an increase in TRIM67 expression
twenty-four hours and seventy-two hours post infection. Based on these observations, we hypothesize that the transcriptional response of TRIM67 to immune stimulation is conserved across vertebrate species. To address this hypothesis, an in
vivo zebrafish embryo model and an in vitro mammalian cell culture system was employed. TRIM67 transcript levels were
measured using quantitative RT-PCR in zebrafish embryos after exposure to immune agonists as well as J774.2 murine
macrophage-like cells after exposure to lipopolysaccharide (LPS). An increase in Trim67 transcript levels was observed in
response to immune agonists in zebrafish embryos at 36 hours, which provides evidence to support pursuing additional time
points. Additional preliminary data suggest there is a decrease in TRIM67 transcript levels in response to LPS stimulation
by murine macrophages. Although changes in transcript levels appear conserved across zebrafish and Atlantic salmon, these
data suggest it may not be conserved in mammals. Knowledge gained by determining whether TRIM67 plays a role during
infections is relevant to veterinary and human medicine.
Research Grant: This work was funded by a grant from the NCSU College of Veterinary Medicine.
Student Support: Veterinary Scholars Program and Fund for Discovery
176
Dysbiosis and bile acid dysmetabolism in dogs with steroid responsive enteropathy
Hannah Klein, Blake Guard, Julia Honneffer, Michelle Jonika, Jan Suchodolski
Texas A&M University CVM, College Station, TX
Chronic enteropathy (CE) is a common gastrointestinal disease that affects all dog breeds. Previous studies have indicated
that dogs with chronic enteropathy have a microbiota imbalance which may have an impact on the severity of the disease.
The purpose of this study was to characterize long term dysbiosis and functional changes in dogs with steroid responsive
CE (SRE). We compared the dysbiosis index and fecal bile acid concentrations (BA) between healthy and diseased dogs.
Fecal samples were collected from healthy dogs (n=20) and dogs with SRE (n=9). Fecal BA were analyzed by gas chromatography coupled with mass spectrometry and the dysbiosis index was evaluated by combining the qPCR values for E. coli,
Streptococcus, Turicibacter, Faecalibacterium, Fusobacterium and Blautia. Healthy dogs had a significantly lower dysbiosis index compared to dogs with SRE (P<0.001). The % of primary BA was significantly higher and the % of secondary BA
were significantly lower in the diseased dogs as compared to healthy dogs (P=0.019). However, no significance was seen
when comparing the total BA, secondary BA or primary BA of both groups. The dysbiosis index, the total BA and the total
secondary BA, were significantly different between time of first diagnosis and three weeks later. In conclusion, the use of
the dysbiosis index and bile acid results is helpful in indicating CE in dogs. The dogs with CE had a significant imbalance in
their intestinal microbiota, and in total and secondary BA as compared to healthy dogs. Future studies are needed to evaluate
the manipulation of fecal bile acids as therapeutics in dogs with SRE.
Research Grant: Veterinary Medical Scientist Research Training Program
Student Support: Comp. Gastroenterology Soc. Vet. Student Research Program
Evaluation of vertebral articular process dysplasia and the thoracolumbar myelopathies in pug dogs
Michael J. Kluz, Elizabeth A. Ballegeer, Kathryn M. Winger, and Jon S. Patterson
College of Veterinary Medicine (Kluz), Department of Small Animal Clinical Sciences (Ballegeer, Winger), Department
of Pathobiology and Diagnostic Investigation (Patterson), Michigan State University, East Lansing, MI
Hind limb weakness and ataxia is considered the second most important health concern in pugs. Signs can progress to paraplegia and include urinary and/or fecal incontinence. Multiple pathologies, including intervertebral disc disease (IVDD) and
spinal arachnoid diverticulum (SAD), have been implicated in the condition broadly termed “Pug Myelopathy.” Caudal articular process dysplasia, a vertebral defect commonly found in pugs, is suspected to cause spinal instability and play a role
in the development of spinal cord lesions in the T3-L3 region. Pugs presenting to the Michigan State University Veterinary
Teaching Hospital with hind limb weakness and/or ataxia referable to the T3-L3 spinal cord were enrolled in our study. Each
patient underwent computed tomography (CT) and magnetic resonance imaging (MRI). Imaging results were analyzed for
a correlation between vertebral caudal articular process dysplasia location and site(s) of spinal cord lesions. Although cord
lesions were most prevalent in the T11-L1 segments, there was no correlation between the sites of caudal articular process
dysplasia and spinal cord damage in the T9-L3 region. Disease progression was analyzed in terms of survival time and time
to onset of paraplegia, urinary incontinence and fecal incontinence. We established that survival time was not significantly
different among diagnostic categories and development of secondary signs was not linked to a particular spinal cord diagnosis; however, fecal incontinence was more prevalent than urinary incontinence or paraplegia.
Research Grant: Michigan Animal Health Foundation and the Pug Dog Club of America
Student Support: Michigan State University College of Veterinary Medicine and Graduate School Fellowship Funds
177
Urine for a surprise: Evaluating the use of expanded quantitative urine culture on feline samples
Emily T. Knebel, Stephen, D. Cole, Shelley, C. Rankin
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA
The microbial flora of the urinary bladder of humans has recently been characterized using an Expanded Quantitative Urine
Culture (EQUC) method, suggesting the presence of a urinary microbiome. Further studies have addressed the role of the
microbiome in urogenital tract disease. Feline lower urinary tract disease (FLUTD) encompasses a common, but very complex spectrum of diseases including urinary tract infections (UTI), urolithiasis and feline idiopathic cystitis (FIC). FIC is
clinically indistinguishable from UTI; however, standard urine culture is negative for FIC patients. The EQUC method has
been shown to enhance the detection of microorganisms by using increased volumes of urine with a variety of culture media
and atmospheric conditions. The aim of this study was to use the EQUC method to explore FLUTD. EQUC was used to
culture urine obtained via cystocentesis from two clinically distinct groups of patients (FLUTD and non-FLUTD cats). Over
the study period, 38 urine samples were available, but 7 were excluded by diagnosis of UTI on standard culture. In total 11
samples were categorized as FLUTD and 20 were categorized as non-FLUTD based on the final diagnosis in the medical
record. EQUC testing was positive for 10/11 (91%) FLUTD and 10/20 (53%) non-FLUTD patient samples. Coagulase-negative Staphylococcus species predominated in both groups (13/24 [54%] of non-FLUTD isolates, and 7/15 [47%] of FLUTD
isolates). This proof-of-concept study showed that EQUC can be used to isolate living organisms from feline urine samples
that are negative by standard culture.
Genetic knockout screens identify host genes required for zika virus replication
Kristi Kobluk, Yaw Shin Ooi, Jonathan Diep, Karim Majzoub, Andreas S. Puschnik, Caleb D. Marceau, and Jan E. Carette
College of Veterinary Medicine, Iowa State University (Kobluk), Ames, IA Department of Microbiology
and Immunology, School of Medicine, Stanford University (Kobluk, Ooi, Diep, Majzoub, Puschnik, Marceau, Carette),
Stanford, CA
Zika virus is a positive sense RNA enveloped arbovirus that belongs to the family Flaviviridae. Zika virus is the first identified Flavivirus that is sexually transmitted and linked to congenital defects. Currently, there are no effective anti-virals or
vaccines against zika virus. Previously, haploid genetic screen technology was used to identify host genes for multiple medically important viruses. We aim to use a similar approach to identify important host factors in zika virus infection that could
be potential drug targets. Genomic scale haploid genetic screens identified host proteins involved in zika virus infection.
Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)/Cas9 nuclease technology was used to generate host
knockout subclones to validate the hits recovered from the haploid genetic screens. Hap1 knockouts were verified through
genotyping to ensure the host protein was mutated. To validate the effects of the host gene knockout on zika virus infection,
assays will be performed. The results will be scored by plaque assays, RT-qPCR, and immunofluorescence assays. The
plaque assays will be used to determine zika virus titer of infectious particles. RT-qPCR will be used to detect the amount
of RNA viral abundance. Immunofluorescence assays will be used to detect the presence of viral protein. By identifying
host proteins that zika virus requires for infection, this research can provide the foundation for creating effective anti-virals
against zika virus.
Research Grant: NIH Directors New Innovator Award
178
The effect of oral sodium bicarbonate “milkshake” on serum total carbon dioxide concentration (TCO2) in horses
Kelsey Koenig, Frank Andrews, Michael Keowen, Travis Miller, Stephen Gaunt, Levent Dirikolu
Equine Health Studies Program, Department of Veterinary Clinical Sciences, (Koenig, Andrews, Keowen, Miller), Clinical Pathology Laboratory, Department of Pathobiological Sciences (Gaunt), and Equine Medication Surveillance Laboratory (Dirikolu), Louisiana State University, School of Veterinary Medicine, Baton Rouge, LA.
During strenuous and intense exercise, lactic acid builds up in the blood leading to poor performance. To buffer lactic acid
and improve performance, a “milkshake” containing sodium bicarbonate (500 grams) is given to horses 2-6 hours before
races. Sodium bicarbonate is thought to buffer lactic acid in the blood, leading to increased performance by preventing
fatigue. Because of its performance enhancing effects, administration of sodium bicarbonate is controversial and illegal in
horse racing. The purpose of this study was to determine if sodium bicarbonate administered via nasogastric tube (NG) to
Thoroughbred horses increased serum TCO2 concentration, an indicator of buffering capacity. Four Thoroughbreds were
studied in a 2x2 crossover design. The horses were housed in stalls during the study. Two horses were randomly selected to
receive a sodium bicarbonate milkshake (500g in 2 liters of water, via NG tube). During period 1, two horses received the
bicarbonate and the other two received no treatment. During period 2, treatments were switched. There was a one week wash
out between periods. Blood samples from the horses were collected before treatment, then hourly for eight hours, and then
24, 48, 72 and 96 hours after treatment in tubes containing no anticoagulant. Blood samples were centrifuged at 1,500g for
15 minutes and serum submitted in the same tubes (without removing the tops) to the clinical pathology laboratory. TCO2
was measured in quadruplicates using a chemistry analyzer (Beckman Coulter A680 analyzer). We expect to find serum
TCO2 significantly increased in the bicarbonate-treated horses, which could enhance performance during exercise.
Research Grant: Department of Comparative Biomedical Sciences
Student Support: Merial Summer Scholars Program, Louisiana State University, School of Veterinary Medicine
Is dietary exposure to bisphenol A causing phenotypic changes in dogs?
Zoe L. Koestel, Robert C. Backus, Kaoru Tsuruta, Juliana Amorin, Savannah Smith, Sarah A. Johnson, Angela B. Javurek,
Kurunthachalam Kannan, Mark R. Ellersieck, Cheryl S. Rosenfeld
Bond Life Sciences Center (Rosenfeld), Department of Veterinary Medicine and Surgery (Backus and Tsuruta), University
of Missouri, College of Veterinary Medicine, Columbia, MO
Bisphenol A (BPA) is a widely prevalent chemical found in common products, including baby bottles, canned items, and
drink packaging. It is an endocrine disrupting chemical (EDC) that is ubiquitous in the environment, ensuring exposure for
generations to come. BPA acts as a weak estrogen and has the potential to cause an array of phenotypic abnormalities such as
modulations in DNA methylation, reproductive function, fetal development and cognitive ability. Animals may be exposed
to BPA when ingesting food products, especially canned items. Past studies have examined effects of non-dietary exposure
to BPA in rodents, fish and turtles. To our knowledge, BPA effects that may result in dogs after consuming canned dog food
lined with BPA-containing material have not been reported. To address this critical gap in knowledge, dogs were fed one of
two different canned diets over a 14 day period. One was lined with material containing presumably scant amounts of BPA
and the other was lined with material containing presumably higher amounts of BPA. Changes in CBC/general chemistry
panel, fecal microbiome and DNA methylation before and after the 14 day period are currently being assessed and correlated with internal concentrations of BPA. Additionally, studies are underway to determine the presence of BPA in the can
lining and BPA concentration in the dog foods. These studies will provide insight as to whether there is cause for concern
in feeding dogs canned diets lined by BPA-containing material and, by translation, humans. Such information may be vital
to veterinary clinicians, nutritionists, and owners in terms of making educated decisions on which dog food minimizes BPA
exposure.
Research Grant: Mizzou Advantage Grant
Student Support: Morris Animal Foundation Student Scholar
179
Evaluation of bone marrow aspiration methods to increase yield and reduce time for therapeutic dose expansion
Rosalind Kopp, Taralyn McCarrel, Kelly Kirkendall-Merritt
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, University of Florida,
Gainesville, Florida
Musculoskeletal injuries in the horse are among the most common applications of adult mesenchymal stem cell (MSC) therapy as well as an active area of research for the benefit of the equine and human patient. One of the major limiting factors
for the utility of MSC therapy is the time required for isolation and culture expansion of an adequate dose resulting in delay
of MSC injection or cell based surgeries for 3 to 4 weeks. Collection of larger numbers of MSC at the time of aspiration
will require fewer cell doublings to achieve therapeutic doses and lessen the current delay in therapy. The objective of this
study was to compare three different sternal bone marrow aspiration techniques for the purpose of mesenchymal stem cell
isolation and culture expansion to determine which technique results in the greatest MSC yield. Six adult Thoroughbred
horses of mixed gender will have bone marrow collected using two different techniques from the fifth sternebra at 1-month
intervals. The number of MSC collected at the time of bone marrow aspiration and the time to achieve a therapeutic dose of
cells will be compared between groups to determine the optimum technique for bone marrow collection from the sternum.
We hypothesize that bone marrow aspiration needle type and technique can be optimized to increase the initial yield of
MSC, thereby reducing culture time and passage number to achieve clinically relevant doses of cells. We propose that a new
multi-site aspiration technique using a fenestrated bone marrow biopsy needle will be clinically feasible and result in greater
stem cell numbers in fewer passages in comparison to single site aspiration with fenestrated bone marrow biopsy needles.
Student Support: Merial Veterinary Research Scholars Program
Rates and Risk Factors for Surgical Site Infections at the Ohio State University Veterinary Medical Center
Caroline Kramer, Joshua Daniels
Department of Veterinary Clinical Sciences (Daniels), The Ohio State University College of Veterinary Medicine,
Columbus, Ohio
Surgical site infections (SSIs) are among the most devastating complications of surgical procedures, resulting in prolonged
pain and recovery time, increased expenses, subsequent procedures, and occasionally death. It is therefore preferable to
prevent SSIs than to treat them. Furthermore, it is more difficult to treat infections caused by antibiotic-resistant bacteria,
which are the cause of many SSIs. Understanding the factors that contribute to SSIs is key to decreasing their prevalence.
The prevalence of SSIs in canine Tibial Plateau Leveling Osteotomies (TPLOs), a common orthopedic procedure, has been
reported to range from 3% to 13%. This retrospective study seeks to determine the prevalence and risk factors for SSIs in
TPLOs performed at the OSU Veterinary Medical Center. Medical records from canine patients (n=865) that underwent
TPLOs between 1/1/2012 and 12/31/2014 were analyzed and evaluated. Data collected (36 variables) included patient demographics, clinical history, anesthesia duration, type of procedure (arthroscopy or arthrotomy, as well as any additional
procedures performed), postoperative antibiotics prescribed, postoperative followup, and diagnosis and treatment of postoperative infections. Of the 865 cases evaluated, 147 cases were lost to followup and 51 surgical site infections were recorded.
The prevalence of SSIs, not including the 147 TPLOs lost to followup, was 7.1%. This study will greatly assist the OSU
VMC in monitoring the prevalence of SSIs and implementing new procedures and policies to lower the occurrence of these
devastating complications.
Student Support: Dr. Michael Rohovsky Fellow
180
Contraception of feral cats with phage-based vaccines targeting gonadotropin releasing hormone (GnRH)
Carol Jeanne Kraneburg, Aime Johnson, Anna Cochran, Jessica Cannon, James Wright, Carla Barstow,
and Tatiana Samoylova
Scott-Ritchey Research Center (Kraneburg, Cochran, Cannon, Samoylova), Department of Clinical Sciences (Johnson,
Barstow) and Department of Pathobiology (Wright, Samoylova), College of Veterinary Medicine, Auburn University,
Auburn, AL
Overpopulation of feral animals, including cats, is a growing concern in the United States and in many other parts of the
world. It was estimated that there are 70 million feral cats in this country alone. Controlling animal populations with anti-fertility vaccines is one of the most promising approaches. The focus of our research is on development of anti-fertility
vaccines that are composed of whole phage particles carrying peptides with contraceptive properties for use in feral animals. The vaccines are designed to stimulate potent antibody responses against GnRH, a master reproductive hormone.
Anti-GnRH antibodies inactivate endogenous GnRH that in turn causes reduced release of gonadotropic hormones leading
to gonadal atrophy in adult animals or lack of development in sexually immature animals. Previously, several phage-GnRH
constructs with potential contraceptive properties were generated via selection from a phage display library. When tested
in mice, such phage-GnRH constructs stimulated production of anti-GnRH antibodies that resulted in suppression of serum
testosterone, a major indicator of impaired fertility. The goal of the present study is to test these phage-GnRH vaccines for
contraceptive potentials in cats. Five domestic male cats (8-9 months old) were characterized as to their reproductive parameters and then immunized with a phage-GnRH vaccine. Blood and semen samples as well as testicular size data were
collected during a 4-week period post-immunization. GnRH antibodies and testosterone in cat serum, testicular volume,
and quality and quantity of sperm are being evaluated. To assess levels and duration of the immunization-associated changes, the experiment will continue for an additional five months.
Research Grant: Auburn University Intramural Grants Program, Auburn, AL.
Student Support: Merial Summer Program for Veterinary Scholars and Scott-Ritchey Research Center.
Using Cold Atmospheric Plasma as an Alternative Treatment for Chronic, Antibiotic Resistant Wounds
Kristin Kuball, Seshagiri Nandula, Chi Phan, Vighneswara Siva Santosh Kumar Kondeti, Peter Bruggeman,
Ryan Hunter and Jennifer Granick
Department of Veterinary Clinical Sciences (Kuball, Nandula, Granick), Department of Mechanical Engineering
(Kondeti, Bruggeman) and Department of Microbiology (Phan, Hunter), College of Veterinary Medicine,
University of Minnesota, St. Paul, MN
The healing and management of chronic wounds is a challenge in both human and veterinary species. Persistent wounds can
be complicated by bacterial infections and many become progressively resistant to antibiotic therapy. Commonly methicillin- resistant Staphylococcus aureus (MRSA) and multi-drug resistant Pseudomonas aeruginosa (MDRPA) are implicated;
these bacteria are capable of forming biofilms that are not resolved by standard treatments. We explored a new therapy, cold
atmospheric plasma (CAP), which we hypothesized has bactericidal effect and the ability to diminish biofilm integrity. CAP
is a partially ionized gas composed of reactive oxygen (ROS) and nitrogen species (RONS), which creates a hostile environment for bacteria while stimulating host cell growth. Our initial studies involved optimizing biofilm growth using MRSA
strain USA300 and MDRPA strain PA14 in vitro followed by treatment with CAP for further microscopic and bioluminescent observations regarding the bactericidal effect and biofilm stability. Previously optimized conditions of a 5-minute CAP
treatment time at a 6mm distance from the biofilm were used. Our preliminary data suggests that under these circumstances,
there is an improvement in the bacterial biofilms not seen with conventionally used antibiotics.
Student Support: National Institutes of Health T35 Training Grant
181
Determining improved prognostic indicators in equine large colon volvulus
Cecilia R Kucera, Laura Stranahan, Liara M Gonzalez
Department of Clinical Sciences, College of Veterinary Medicine, North Carolina State University, Raleigh, NC
Large colon volvulus (LCV) is a devastating disease in horses that necessitates surgical intervention. Stem and progenitor
cells play a critical role in epithelial repair following ischemic events in the intestine. Protein biomarkers are used to identify
these progenitor cells in the colon. This study aims to determine if a critical number of intestinal progenitor cells can predict
tissue viability and outcome in cases of LCV. The objectives were to 1) identify intestinal progenitor cells using biomarkers:
proliferating cell nuclear antigen (PCNA), sex determining region Y box 9 (SOX9), phosphorylated histone 3 (PH3), and
Ki-67, 2) define cutoff values for critical numbers of positive cells and 3) determine if case prognosis is associated with
cutoff values. Twenty-three cases of LCV $360o with pelvic flexure biopsy and that recovered from surgery were included
in this study. Thirteen of 23 (57%) horses survived to discharge. Immunohistochemical analyses of formalin fixed biopsies
were performed and positive cells per crypt were counted. The optimal cutoff value for positive cells was determined using
receiver operator curves. The association of cutoff values with survival was determined using a Fisher’s exact test. A cutoff
value of <2.1 PH3 positive cells per crypt correctly predicted death with 100% sensitivity (95% CI; 69.15%-100%) and
84.62% specificity (95% CI; 54.55%-98.08%). LCV cases with <2.1 PH3 positive cells per crypt were 96.6 times more likely to die (95% CI; 4.142-2253 and p<0.0001). Biomarkers PCNA, SOX9, and Ki-67 were not predictive of case outcome.
PH3 immunohistochemical analysis provides accurate prognosis in cases of LCV in the post-operative period.
Research Grant: NIH SERCA [or] K01OD0199
Student Support: NIH Training Grant T35OD011070 [or] George H. Hitchings New Investigator Award
Ataxic Horses Display Quantifiable Gait Abnormalities Determined By Videography
Anisha Kumar, Megan E. Aanstoos, L. Ray Whalen, Yvette S. Nout-Lomas
Department of Clinical Sciences, College of Veterinary Medicine and Biomedical Sciences, Colorado State University,
Fort Collins, CO
Scoring severity of neurological gait abnormalities in horses has poor intra-observer reliability. There is a need to develop
unbiased measures of ataxia in horses. The objective of this study was to analyze footfall patterns and quantify gait abnormalities in sound and ataxic horses. We hypothesized that ataxic horses would have a different footfall pattern and more
observable and quantifiable gait abnormalities compared to sound horses. Twenty-three adult horses were used; 11 were
ataxic based on neurologic examination and 12 were sound. From video footage, lateral views were truncated to 40-60 consecutive steps at the walk and gait patterns were analyzed by one observer. Data were compared using Fisher’s Exact Test
and unpaired T-test. Data is reported as average6 standard error and significant is set as p<0.05. The study was approved
by the CSU IACUC. All horses exhibited a 4-beat lateral gait as seen in other quadrupeds. Although all horses displayed
gait abnormalities, in particular interference, significantly more ataxic horses displayed toe dragging (11/11, p= 0.001) and
abnormal limb movement from neutral (6/11, p= 0.03) compared to sound horses (4/12 and 1/12, respectively). Further,
ataxic horses exhibited toe dragging in a significantly higher percentage of steps (26.466.37%) when compared to sound
horses (1.1760.52%; p<0.001). Ataxic horses also have more abnormal limb movements from neutral when compared to
sound horses. Although these results are not surprising from a clinical perspective, these gait abnormalities have not before
been quantified in ataxic horses and our findings suggest that quantifiable parameters could be of use in development of a
more objective grading scale for ataxia in the horse.
Student Support: USDA-NIFA Animal Health & Disease Research Program Funding 2016-36100-06008
182
Optimization of ultrasound settings for better determination of cystolith size in vitro
Lauren E Kustasz, John M Kruger, Nathan C Nelson
College of Veterinary Medicine (Kustasz, Kruger, Nelson), Michigan State University, East Lansing, MI; Department of
Small Animal Clinical Sciences (Kruger, Nelson), Michigan State University, East Lansing, MI
Accurate estimation of cystolith size is a critical factor in assessing the biological behavior of urocystoliths, their response
to dissolution therapy, and their potential for removal by minimally invasive procedures. A previous study found that ultrasound, using a curved array transducer at a frequency of 10 MHz, overestimated cystolith size compared to other imaging techniques. The purpose of this study was to further evaluate ultrasonography as an imaging technique for measuring
cystoliths, comparing transducer types, different frequencies, and the use of tissue harmonics imaging and spatial compound
imaging using an in vitro bladder phantom model. Thirty cystoliths were imaged using combinations of the ultrasound variables mentioned. The accuracy of cystolith measurement was determined by taking the difference between the measurement
obtained from the ultrasound image and the true size of the cystolith determined by a digital caliper. The accuracy of the
measurements obtained from the linear transducer was significantly greater than the accuracy of the measurements obtained
from the curved array transducer (p < 0.05). Measurements from the linear transducer showed a significant decrease in accuracy of cystolith size estimation when spatial compound imaging was used (p < 0.05 ). Independent of actual cystolith size,
the linear transducer tended to overestimate cystolith size by 0.16 cm on average and the curved transducer overestimated
by 0.43 cm on average. Subjectively, there were more artifacts seen in the images taken with the curved transducer and,
especially with smaller cystoliths, these artifacts superimposed the cystoliths making them difficult to measure.
Bathilda Lake, Kurt Zimmerman, Sharon Witonsky, W. Kent Scarratt, Harold C. McKenzie, III, Tony Huffman,
Stephanie Todd, and James Holt
Biomedical Sciences and Pathobiology (Lake, Zimmerman), Large Animal Clinical Sciences (Witonsky, Scarratt,
McKenzie), Equine Field Services (Huffman), Infectious Disease Research Facility (Todd), Virginia Maryland College
of Veterinary Medicine, Virginia Tech, Blacksburg, VA; New Holland Sales Stable (Holt), New Holland, PA
Differentiating between horses with regenerative and non-regenerative anemia can assist the clinician in determining the
cause and treatment. However, evaluating regenerative status in equine anemias is difficult because horses do not release
reticulocytes into circulation. The purpose of this pilot study was to determine if there is a relationship between intracellular
erythrocyte potassium concentration (K+) and surface receptor transferrin expression. It has been shown that transferrin expression increases with erythrocyte immaturity. The hypothesis of this study was that intracellular erythrocyte K+ would increase in positive correlation with transferrin expression; signifying the utility of intracellular erythrocyte K+ as a biomarker
for identifying equine regenerative anemias. The following were measured in 13 horses: blood packed cell volume (PCV) by
microhematocrit tube centrifugation of heparinized blood; serum and lysed heparinized blood K+ by ion-selective electrode
method; mean cell K+ concentrations (MCKC) normalized for statistical analysis; and transferrin by commercial ELISA assay on lysed heparinized blood. Results included: PCV range 19-54%; MCKC range 63.4-92.4 mEq/L; and transferrin range
-43.7-63.7 pg/L. Data were found to be normally distributed. Pearson correlation results were significant for PCV-MCKC
(coefficient 0.599, P-value 0.018) and approached significance for MCKC-transferrin (coefficient -0.471, P-value 0.077).
However, no association was found between PCV-transferrin (coefficient 0.059, P-value 0.835). These findings do not support the hypothesis that MCKC increases in association with surface receptor transferrin expression in equine erythrocytes.
Research Grant: Virginia Maryland College of Veterinary Medicine
Student Support: None.
183
Evaluation of the roles of plasmin and CCL20 on nerve damage in an inflammatory M. leprae infection model
Holly Lamb, Vilma T. Marks, Angelina T. Deming, & Linda B. Adams
DHHS, HRSA, Healthcare Systems Bureau, National Hansen’s Disease Programs, Baton Rouge, Louisiana
Leprosy, or Hansen’s disease, is a chronic infection caused by the acid-fast bacterium Mycobacterium leprae that can lead
to deformities and disabilities if not treated early and appropriately. While most of the deformities associated with Hansen’s
disease are due to peripheral nerve infiltration and damage, it has been difficult to reproduce this neuropathy in existing
murine models. With the crossing of the nitric oxide synthase knockout (NOS2-/-) mouse and the IL- 10 knockout (IL-10-/-)
mouse, a new double knockout murine strain (10NOS2-/-) has been established which exhibits an exaggerated inflammatory
response with infiltration of CD4+ T cells into damaged nerves. A closer look at the 10NOS2-/- mice revealed a >100 fold
increase in the expression of the inflammatory chemokine, CCL20. Other studies have shown that binding of mycobacterial
antigens to plasminogen can activate its conversion to plasmin, and plasmin can induce CCL20 at sites of inflammation. It
has also been demonstrated that CCL20 is upregulated at the site of CD4+ T cell entry into the central nervous system in
an experimental model for multiple sclerosis. The purpose of this project will be to look more closely at the interactions
between M. leprae antigens, plasmin, and the inflammatory chemokine, CCL20, to ascertain whether these interactions may
play a role in the neuropathy associated with Hansen’s disease.
Research Grant: Study funded in part by HRSA, American Leprosy Missions, Society of St. Lazarus of Jerusalem,
and Heiser Foundation.
Characterization of a co-culture model for observation of Francisella tularensis infection
Casey Landis, Willie Collins, Jerry Ritchey, Ashish Ranjan, Jerry Malayer
Department of Physiological Sciences and Department of Pathobiology (Ritchey), Center for Veterinary Health Sciences,
Oklahoma State University, Stillwater, OK.
Francisella tularensis is the etiologic agent of Tularemia, a potentially fatal disease of both animals and humans. As an
intracellular bacterium, F. tularensis has evolved the means to evade processing by cells of the immune system, processes
that have been studied in mouse models, but there is a lack of studies concerning the infection process in human cells. We
hypothesize that a co-culture of macrophages and hepatocytes can provide an adequate model for infection. THP-1 monocytes, and HEP2 cell lines were used for a human co-culture. For comparison J774a.1 and AML-12 cell lines were used
for a murine co-culture. The live vaccine strain (LVS) of F. tularensis expressing green fluorescent protein (GFP) was used
to infect the co-cultures for observation. The co-cultures were characterized by confocal microscopy, flow cytometry and
qRT-PCR. For microscopy, cells were stained with DAPI and cytoID to visualize the genetic material and cell phagosomes,
and the GFP F. tularensis. Flow cytometry was used to quantify cells at intervals over a 48 hour period to evaluate infection
status and expression of MHC II, CD206, CD119, and CD89 cell surface markers. One-step qRT-PCR was used to identify
and measure expression of cytokines and cell markers in response to infection. Preliminary results indicate clear differences between infected and non-infected host cells. Confocal microscopy visualized the process of infection, replication, and
release. Flow cytometry revealed a decrease in the cell surface markers, supporting the efforts of F. tularensis to remain
undetected by the immune system. Results to date confirm the co-culture model as an adequate representation for a living
animal in observation of F. tularensis infection.
Student Support: We acknowledge the support of the Oklahoma State CVHS and the Summer Scholars Research Program
184
Exploration of tongue muscle pathology in canine degenerative myelopathy
Victoria Landreth, Shelby Mancini, Joan R. Coates, and Teresa Lever
Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, Missouri
Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disorder in people that primarily affects motor neurons,
resulting in progressive muscle atrophy and paralysis. The pathogenesis of this disease is largely unknown and difficult to
study in ALS patients. We propose that canine degenerative myelopathy (DM) may be a suitable animal disease model, given that DM similarly results in adult-onset, progressive neuromuscular degeneration. In addition, DM and familial ALS are
linked to mutations in the superoxide dismutase 1 gene (SOD1). In ALS, respiratory and swallowing impairment are hallmark features, both seen with tongue muscle pathology, particularly of the genioglossus muscle. Atrophy of tongue muscles
has not yet been investigated in DM, although impaired tongue movement has been reported in later stages of the disease.
The genioglossus is normally composed predominantly of type 2 myofibers; however, based on findings that intercostal
muscles of DM-affected dogs developed a type 1 myofiber predominance, we expect to find a similar pattern in the genioglossus. To see if this is the case, we are investigating post-mortem genioglossus samples from age- and breed size-matched
controls (n=11) and DM-affected dogs spanning all disease stages (n=16). Cross sections of paraffin-embedded samples will
be stained using histochemical and immunohistochemical methods for characterization of markers of muscle degeneration.
Compared to controls, we hypothesize the DM-affected genioglossus will have: 1) reduced abundance of type 2 fibers, 2)
SOD1 protein aggregation, and 3) altered myofiber diameters. Results of this study may provide novel evidence of tongue
muscle pathology in DM to parallel findings in ALS patients.
The effects of pet ownership on academic achievement among veterinary students at St. George’s University
Maya Lantz, and Satesh Bidaisee
College of Veterinary Medicine, St. George’s University, Grenada, West Indies
An extensive amount of research has been done studying pet ownership and its effects on health and education, but to date
no research has addressed how pet ownership affects the academic achievement of veterinary students. The present study
examined the effects of pet ownership of veterinary students at St. George’s University. All participants completed a survey
and answered questions about demographics, pet ownership, academics and social life. A total of 53 responses were collected; 96.2% (N=51) females and 3.8% (N=2) males, ages ranged from 20 to 41 with an average of 26.39 years. Pet owners
made up 92.5% (N=49) of respondents. The average number of pets owned per pet owner is 2.49 pets with a range of 1 to
8 pets. Average reported stress levels of pet owners was 7.29 and 7.25 for non-pet owners using a Likert scale from 1-10.
When asked if having their pets with them decreased their overall stress 69.8% (N=37) said yes and zero responded with
no. Results show that pet owners, students who own pets (and have their pet with them) while in veterinary school have a
perceived decrease in stress due to the human-animal bond between them and their pets.
Student Support: Merial and Island Veterinary Scholars Program
185
Discovery of a genetic mutation that causes sudden cardiac death in toy Manchester terriers
Shannon N. Larrabee, Paula S. Henthorn, Nicole M. Tate, Katie M. Minor, Shannon A. Martinson, Eva Furrow
College of Veterinary Medicine, University of Minnesota, St. Paul, MN, USA (Larrabee, Tate, Minor, Furrow),
School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA, USA (Henthorn), University of Prince
Edward Island, Charlottetown, Prince Edward Island, Canada (Martinson)
A juvenile cardiomyopathy has previously been reported in toy Manchester terrier (TMT) dogs. Affected dogs experience
sudden death at a young age (10 weeks to 1 year). There are little to no preceding clinical abnormalities, with the exception
of cryptorchidism in males. Necropsy findings are consistent with a dilated cardiomyopathy type disorder, with myocardial
degeneration and fibrosis on histopathologic review. The aim of this project was to perform a genome-wide association
study (GWAS) and follow-up sequencing to locate the genetic mutation responsible for juvenile cardiomyopathy in the
TMT. Twelve cases (puppies with histopathology-confirmed cardiomyopathy) and 36 controls (healthy adults >2 years old)
were included in the study. The GWAS identified a highly significant association on chromosome 27 (p-value of 2.7 x 10-15).
Positional candidate genes included two genes encoding cardiac potassium channels that have been linked to sudden death
or adult DCM in people. Using Sanger sequencing, one gene was found to have a missense mutation in a highly conserved
amino acid that was predicted to be deleterious. Mutation testing confirmed the mutation was present in a homozygous
state in all affected individuals (12 original cases plus 2 additional). Samples from 102 healthy TMT dogs were also tested.
A single 8 week old puppy was found to be homozygous for the mutation; this dog was cryptorchid, and thus suspected to
be subclinically affected. The other 101 dogs were either clear or carriers of the mutation (0.08 mutant allele frequency).
In summary, we discovered an autosomal recessive mutation in a cardiac potassium channel that is associated with a fatal
juvenile cardiomyopathy in TMT dogs.
Research Grant: Private donations from Manchester terrier breeders and owners
Student Support: Merial Limited and the College of Veterinary Medicine, University of MN
The architecture of the Flavivirus 3’ untranslated region, pathogenic RNA production, and viral cytopathicity
Hannah M. Laurence, Benjamin M. Akiyama, Jeffrey S. Kieft
Howard Hughes Medical Institute (Laurence), UC Davis School of Veterinary Medicine, Davis, CA (Laurence),
Department of Biochemistry & Molecular Genetics, University of Colorado School of Medicine, Aurora, CO (Laurence,
Akiyama, Kieft)
Flaviviruses (FVs), which include Zika, Dengue, West Nile, and Yellow Fever, are growing global threats with no available
anti-viral therapy. In studied Flavivirus (FV) infections, a viral non-coding RNA is produced to high levels. This subgenomic Flaviviral RNA (sfRNA) has been directly linked to viral cytopathicity. sfRNAs are produced when a host cell’s exoribonuclease, Xrn1, degrades the viral RNA in the 5 prime to 3 prime direction, but stalls at specific sites in the viral RNA’s 3
prime untranslated region (UTR) and cannot complete degradation. The structures that halt Xrn1 are Xrn1 resistant RNAs
(xrRNAs) and the multiple xrRNAs in each FV result in production of sfRNAs of specific lengths and quantities. Biochemical techniques have been used to investigate the kinetics of Xrn1-mediated RNA degradation in the context of different
FV RNA constructs, including Dengue, West Nile, and Zika virus. To examine this, we developed an in vitro assay using
radiolabeled RNAs to quantitate resistance to Xrn1 over time and analyzed the results using denaturing polyacrylamide gel
electrophoresis to identify degradation products based on relative size. We are using virological and cell culture techniques
to examine the cytopathic effects of the major FVs in both mammalian and mosquito cell lines. Viral effects differ in host
versus vector cells, as vectors remain unaffected by overt clinical signs. The goal is to understand how the identity, pattern,
and location of xrRNAs produce specific sfRNAs, to determine how Zika virus biochemistry compares to known FVs, and
ultimately how the pattern of sfRNAs affects the cytopathicity of a Flavivirus to identify targets for therapeutic intervention.
Research Grant: R01 GM081346
Student Support: Howard Hughes Medical Institute Medical Research Fellowship
186
Yeast surface display library to yield prospective homing protein to the ischemic myocardium
Danielle E. LaVine, Ke E. Cheng, Balaji M. Rao, Carlos Cruz-Teran, Adam Mischler, Teresa C. DeFrancesco,
Kathryn M. Meurs
North Carolina State University, College of Veterinary Medicine, Raleigh, NC (LaVine, Cheng, DeFrancesco, Meurs),
Department of Chemical and Biomolecular Engineering, North Carolina State University, Raleigh, NC (Rao, Cruz-Teran,
Mischler)
Myocardial infarction (MI) remains a common cause of morbidity and mortality in the United States, affecting approximately 1.5 million humans annually. Current in vivo target therapy to the ischemic heart has been achieved using phage display in
mice. The yeast surface display library (YSD) has yielded additional proteins when compared to phage display. The purpose
of this study was to discover a novel peptide which exhibits preferential binding to the ischemic myocardium using a YSD
library relative to control heart. In vitro peptide selection for ischemic myocardium was accomplished using a YSD library.
Biopanning was performed using a cryopreserved control and MI-induced rat heart sections. MI was induced by ligating the
left anterior descending artery. EBY100 strain yeast were induced with SGCAA media and grown with SDCAA media. Two
negative selections were performed during each round of biopanning. Six rounds of biopanning were performed. Stringent
negative selection (liver, kidney, lung, skeletal muscle, brain) will be performed. After 6 rounds of biopanning we concluded
that our cryopreserved sections yielded non-specific binding to the myocardium of both control and MI induced rats. Our
goal is to run a stringent negative control that will yield a protein that binds specifically to the myocardium. With future
investigations, this peptide may yield a targeted intravascular delivery method for stem cells and other therapeutics which
will provide an attractive alternative to more invasive delivery methods.
Student Support: North Carolina State University, CVM, Fund for Discovery and NIH grant # T35OD011070
Influence of donepezil on the development of Toxoplasma gondii and its potential relationship to Alzheimer’s
Tyler J. Lawnichak, David S. Lindsay
Blacksburg, Virginia
Donepezil hydrochloride (DH) is an acetylcholinesterase inhibitor used for the treatment of mild to severe Alzheimer’s type
dementia. DH is thought to exert its therapeutic effect through the enhancement of cholinergic function by increasing the
concentration of acetylcholine through reversible inhibition of hydrolysis by acetylcholinesterase. Toxoplasma gondii is an
obligate intracellular parasite in the phylum Apicomplexa. Cats and other felids are the parasite’s definitive hosts however,
T. gondii infects a wide variety of warm-blooded intermediate hosts including humans. T. gondii is of special interest for
brain related dysregulation because of its known affinity for brain tissue and its capacity for stable, lifelong infection. T.
gondii has been implicated as a cause of a variety of neuropsychiatric disorders particularly schizophrenia. In recent years,
it has been suggested that a potential relationship with other neurobiological diseases, such as Parkinson’s disease and
Alzheimer’s disease, may also exist. In this study, infection of mammalian host cells with T. gondii and treatment of this
infection with DH was performed to observe the response of T. gondii tissue cyst production and maintenance as well as the
potential relationship this may have to Alzheimer’s patients being treated with DH. Our preliminary studies indicate that a
single treatment with DH may actually increase the number and size of tissue cysts. The early observed effects on tissue cyst
development and maintenance may be relevant in interpreting reports of DH treatment failures in patients with confirmed
Alzheimer’s disease.
Research Grant: NIH 4T35OD011887-10
187
Validation of putative chromosome 14 risk alleles for osteochondrosis in Standardbred horses
Eric J. Ledesma, Megan Darragh, and Annette M. McCoy
Osteochondrosis (OC) is a developmental orthopedic disease characterized by focal failure of endochondral ossification. It
is believed that vascular failure leading to ischemic chondronecrosis results in OC, although the etiology of this pathology is
unknown. Both genetic and environmental factors are believed to play a role in the risk of developing OC. Previously, two
regions of equine chromosome 14 were validated for association with risk of hock OC in Standardbreds. We hypothesize
that specific variants within these regions have a moderate effect on hock OC risk. In this study, specific single nucleotide
polymorphisms (SNPs) located within these regions were assessed for association with OC in an independently sampled
Standardbred population. This population will be prospectively assessed radiographically for OC development, between 2
and 12 months of age, with a definitive OC diagnosis made at >8 months. Primers surrounding 14 SNPs of interest were
designed using the assembly Equus caballus 2.0. Subsequently, NEBcutter v2.0 was used to design restriction enzymes for
genotyping via restriction fragment length polymorphism (RFLP). SNPs for which RFLP could not be designed were genotyped by Sanger sequencing. Simultaneously, candidate genes in these regions were Sanger sequenced to discover novel
SNPs. These will be tested by similar methods in the experimental population. Our long-term goal is to develop a diagnostic
genetic panel to identify at-risk animals, which would allow clinicians and owners/trainers to make management changes
to decrease environmental risk factors and overall risk of developing OC. The diagnostic genetic panel would also allow
more-informed breeding decisions within high-risk pedigrees.
Research Grant: Morris Animal Foundation D16EQ-311 and USDA Hatch Funds
Examination of insectivorous bats (Molossus molossus) in Grenada for trematodes and Neorickettsia DNA
Amber L. Lee, Kathryn E. Gibson
Department of Pathobiology, St. George’s University School of Veterinary Medicine, St. George’s, Grenada, West Indies
Neorickettsia, obligate intracellular bacteria implicated in both human and animal diseases, are the only known bacteria to
utilize a digenetic trematode host to complete their own life cycle. In this study, the life cycle of Neorickettsia risticii and its
trematode host, which utilize an insectivorous bat as the naturally-infected host of the bacterium and definitive host of the
trematode is explored on the Caribbean island of Grenada. There are no previous studies on Neorickettsia or digenetic trematodes that could harbor Neorickettsia in Grenada. Our hypothesis is that trematodes, duodenums, livers and/or spleens from
insectivorous velvety free tailed bats (Molossus molossus) in Grenada contain DNA of N. risticii, or similar species, and
the trematode vector will be identified as belonging to the family Lecithodendriidae. Duodenums, spleens, and livers were
checked for the presence of trematodes and/or Neorickettsia. Morphologically, 23.1% (6/26) of duodenums had identifiable
trematodes. Conventional PCR results utilizing primers for the ITS2 region indicated that 73.1% (19/26) of duodenums had
trematode DNA, which was identified as Lecithodendrium sp. by sequencing. PCR to identify a 382-bp Neorickettsia 16S
rRNA gene fragment yielded a larger (~500-bp) band in 50% (3/6) of trematodes, 10% (1/10) of intestines, and 17% (2/12)
of spleens/livers, which was unidentifiable. While Neorickettsia DNA was not identified, the presence of trematodes from
the family Lecithodendriidae strongly supports the claim that Grenada is a suitable setting for Neorickettsia to survive.
Further studies will have to be conducted to definitively conclude its non-existence on the island and if it has the potential
to cause disease in Grenada.
Research Grant: SGU Small Research Grant Initiative (SRGI) no. 12004
Student Support: Merial Veterinary Scholars Program (MVSP) cosponsored by Merial and St. George’s University
188
Management practices and disease perceptions among Minnesota backyard flock owners following a HPAI outbreak
Meng Li, Larissa Minicucci, Riikka Soininen, Gilbert Patterson, Lou Cornicelli, Wayne Martin.
University of Minnesota, Center for Animal Health and Food Safety, St. Paul, MN (Li, Minicucci, Soininen, Patterson),
Minnesota Department of Natural Resources, St. Paul, MN (Cornicelli) and University of Minnesota Extension, St. Paul,
MN (Martin).
During the 2015 outbreak of highly pathogenic avian influenza (HPAI) in Minnesota, over 9 million birds, 108 farms and
23 counties were affected. The economic impact of the outbreak on the Minnesota economy was estimated to be over 650
million US dollars. However, only one of the 108 infected farms was a backyard flock. Understanding factors that may affect
disease transmission and control among backyard flocks would be beneficial in defining disease risk and prevention strategies amidst a possible future outbreak. A pilot survey project was conducted to better understand flock management and
biosecurity practices, environmental exposures, and owner knowledge of poultry diseases. Additionally, owner perceptions
surrounding HPAI were evaluated. A convenience sample of 244 backyard flocks in Minnesota was selected from a list of
registered egg and poultry producers. A forty-seven-question survey was sent by email to the selected sample. As of July
14th 2016, 29 poultry owners responded, for a response rate of 11.9%. The preliminary results indicate that 62% (18/29)
of owners believe they have adequate biosecurity measures in place to prevent HPAI. Interestingly, 31% (9/29) were not
concerned about HPAI when asked about biosecurity. Few biosecurity measures have been changed by flock owners since
the 2015 HPAI outbreak with only 24% (7/29) of owners making adjustments. The most common change described was
limiting visitor access. This initial data suggests that education efforts may be warranted for backyard flock producers to
raise awareness and understanding of biosecurity practices in relation to diseases such as HPAI.
Research Grant: Minnesota Department of Agriculture as part of the University of Minnesota Response to Avian Influenza
Grant
Student Support: University of Minnesota Center for Animal Health and Food Safety
Characterization of a Sox2-expressing subpopulation of cells in canine lymphoma
Seth Lieberman, Dania Villarnovo, Kristy Richards
Department of Biomedical Sciences, Cornell University College of Veterinary Medicine, Ithaca, NY
Cancer stem cells (CSC) are a subgroup of tumor cells thought to play a critical role in the initiation, metastasis, and chemoresistance of neoplasms. Sox2, a member of the sex determining region Y box family, is mostly found in embryonic stem
cells where it promotes pluripotency and suppresses differentiation. Previous research has shown that Sox2 is overexpressed
in some forms of solid tumors where it has been correlated with a poor prognosis and has served as a marker for CSCs. Our
laboratory is investigating a subpopulation of cells in canine lymphoma patient tissues and cell lines that are positive for
Sox2 by immunohistochemistry on the tissues and immunocytochemistry on the cell lines. We are currently optimizing a
Western Blotting protocol in order to verify Sox2 protein expression in these same cell lines. RT-PCR will also be employed
to test for the presence of Sox2 RNA expression. Finally, detection of the Sox2-expressing cells with flow cytometry will
allow us to isolate these cells in order to investigate which other CSC properties they exhibit. These properties include the
expression of members of the membrane-bound ABC (ATP-binding cassette) family of transporters including ABCB1,
ABCG2, and ABCC1 and by sphere and colony-forming abilities. By further characterizing the CSC properties of Sox2-expressing cells, we aim to identify a subpopulation that may be responsible for causing resistance to therapies and which can
potentially be targeted in relapsed patients.
Research Grant: Dr. Kristy Richards start-up fund from Cornell University
Student Support: Office of Graduate Education at Cornell University College of Veterinary Medicine
189
Two methods of Clostridium difficile toxin purification for detection by MALDI-TOF MS
Dian Dian Lin, Christina R. Wilson, and G. Kenitra Hammac
College of Veterinary Medicine, Purdue University, West Lafayette, IN (Lin, Wilson, Hammac), Department
of Comparative Pathobiology, Purdue University, West Lafayette, IN (Wilson, Hammac), Indiana Animal Disease
Diagnostic Laboratory, West Lafayette, IN (Wilson, Hammac)
Clostridium difficile is an important cause of diarrhea and enterocolitis in horses, pigs, dogs, cats and humans. Two large
exotoxins, toxin A (TcdA), an enterotoxin, and toxin B (TcdB), a cytotoxin, are essential virulence factors and account for
the clinical signs of Clostridium difficile-associated diseases. Since not all C. difficile strains are toxigenic, accurate diagnosis relies on the detection of toxin from fecal samples or bacterial isolates. Previously, our laboratory has assessed the
use of matrix assisted laser desorption ionization - time-of-flight mass spectrometry (MALDI-TOF MS) to efficiently detect
proteolytic digests of toxin A and toxin B from commercially available, purified toxins. In the present experiment, toxins
were purified from two ATCC C. difficile strains by two methods: (1) using a centrifugation filter devise for size exclusion;
or (2) via binding to galactose agarose. Subsequently, masses obtained by MALDI-TOF MS analysis were compared to
expected masses available from an online database and to the masses generated from commercially available, purified toxins. Few matches were found between our results and expected peptide masses of TcdA and TcdB. In addition, the masses
from sample isolates were poorly matched with masses generated from purified, commercially available toxins. Our results
therefore suggest that MALDI-TOF MS may be unsuitable as a method for the diagnosis of C. difficile infections. Further
studies regarding toxin purification methods and MALDI-TOF MS methods of analysis are required.
Progression of vertebral bone pathology in mucopolysaccharidosis VII dogs from birth to skeletal maturity
Megan Lin, Sun H. Peck, Jennifer L. Kang, Neil R. Malhotra, Patricia O’Donnell, Mark E. Haskins, Margret L. Casal,
Lachlan J. Smith
School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA (Lin, O’Donnell, Haskins, Casal),
Departments of Neurosurgery and Orthopaedic Surgery, Perelman School of Medicine, University of Pennsylvania,
Philadelphia, PA (Peck, Kang, Malhotra, Smith)
Mucopolysaccharidosis (MPS) VII (Sly Syndrome), is a genetic lysosomal storage disease characterized by mutations in
the b-glucuronidase (GUSB) gene, which leads to deficient enzyme activity and abnormal accumulation of glycosaminoglycans. Spinal abnormalities are prevalent in human and canine patients. Currently, there is no treatment for the disease.
MPS VII spines exhibit delayed and incomplete cartilage-to-bone conversion in vertebral secondary ossification centers.
Previous research demonstrated that secondary ossification occurs by 14 days-of-age in healthy dogs, but not until 30 daysof-age in MPS VII dogs. At 9 and 14 days-of-age, trabecular bone content is the same in both healthy and MPS VII dogs,
but beyond 30 days-of-age, MPS VII vertebrae exhibit lower trabecular bone volume fraction and mineral density. Based on
these findings, we hypothesize that as age increases, MPS VII dogs will have decreased bone volume and mineral density, as
well as growth plate abnormalities compared to healthy control dogs. Using the naturally-occurring MPS VII canine model,
our objectives are to establish differences in 1) trabecular bone volume and mineral density in vertebral primary ossification
centers and 2) the morphology of vertebral growth plates, including heights, areas, and chondrocyte cell numbers of proliferative and hypertrophic zones between control (heterozygous) and MPS VII dogs. Vertebrae from control and MPS VII dogs
(n=3-5) at 1.5, 3, 6, and 12 months-of-age will be analyzed using micro-computed tomography and histology. The results
will further elucidate the progression and underlying mechanisms of MPS VII spine disease and inform future research for
treatment of skeletal manifestations of all MPS subtypes.
Research Grant: P30 AR050950, P40 OD010939, R03 AR065142
Student Support: NIH/Merial Veterinary Scholars Grant
190
The effects of tongue injection of CTB-SAP on ventral hypoglossal motor neurons: a novel model of ALS
Lori A. Lind, Teresa E. Lever, and Nicole L. Nichols
College of Veterinary Medicine (Lind), Department of Otolaryngology-Head and Neck Surgery, School of Medicine
(Lever), and Department of Biomedical Sciences, College of Veterinary Medicine (Lever, Nichols),
University of Missouri, Columbia, MO
Amyotrophic lateral sclerosis (ALS) is a progressive disorder in which most patients lose the ability to breathe and have
to be placed on a ventilator, and many experience dysphagia which often leads to placement of a feeding tube. Although
transgenic rodents are available for research, they take months to develop ALS and are highly variable regarding type of impairment exhibited. Researchers working on treatments for ALS could benefit from an inducible model in which symptoms
can be quickly and reliably produced. Nichols et al. (2015) developed a model using 25mg cholera toxin B conjugated to
saporin (CTB-SAP) which mimicked aspects of ALS related to respiratory deficits. The purpose of this study is to determine
if CTB-SAP can be used to recapitulate the dysphagia aspect of ALS. We hypothesize that genioglossal injections of CTBSAP will produce targeted cell death in the ventral hypoglossal (XII) nucleus. Anesthetized adult male rats will receive a
tongue injection of either: 1) 25mg CTB-SAP + 20mg CTB; or 2) 20mg CTB unconjugated to 25mg saporin (control). Lick
rate and swallow assays will be conducted to determine if resultant deficits are similar to those observed in ALS mouse
models. Subsequently, XII motor output will be recorded, rats will be perfused, brainstems will be sectioned, and immunohistochemistry will be used to detect and count surviving CTB(+) stained XII motor neurons. We predict that rats injected
with CTB-SAP will have fewer surviving XII motor neurons than controls. Besides being rapidly inducible, this model
should produce animals with none of the other clinical signs associated with ALS, thus making it ideal to study treatments
aimed specifically at dysphagia.
Research Grant: University of Missouri College of Veterinary Medicine Committee on Research (COR) Grant and
Richard Wallace Faculty Incentive Grant
Experimental modeling of the nonspecific protective effects with measles virus vaccination
Sarah C. Linn, Stefan Niewiesk
Department of Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH
The administration of a vaccine can have non-specific effects that are protective against unrelated pathogens in an infant
patient. A recent study in Denmark observed that children whose most recent vaccine was the live measles-mumps-rubella
vaccine had a lower rate of respiratory syncytial virus hospitalization compared to children who had the inactivated DTaPIPV-Hib3 vaccine. In addition, an epidemiological link has been found between measles vaccination and Streptococcus
pneumoniae infection. In our study, we aimed to provide the experimental basis for the nonspecific protective effects of measles virus immunization. S. pneumoniae was grown overnight and OD600 values were used to determine the colony forming
units (CFU)/mL to use for infection. Cotton rats were infected with 10uL or 100uL doses of 5x105 CFU intranasally to assess
bacterial growth in both the nasal passages and the lungs. Nasal washes and bronchoalveolar lavages were performed with
5ug/mL gentamicin in PBS 1 week post infection. No bacterial growth was observed in the lungs but an average of 1.5x105
CFU/mL colonized the nasal passages. To determine the effects of measles vaccination, cotton rats were either not immunized or immunized with measles virus intranasally or subcutaneously and infected with S. pneumoniae at different time
points post-immunization. Bacterial titers from nasal washes 1 week after infection suggests there is a protective effect of
measles virus vaccination 1 week before challenge. Titers from additional time points are still being tested. Potential explanations for the non-specific protective effects of vaccination are cross-reactive immune responses and cytokine secretion as
well as epigenetic changes of the immune response.
Research Grant: Niewiesk Time Release Fund
Student Support: American Veterinary Medical Foundation 2nd Opportunity Research Scholarship
191
Biofilm formation by mastitis-producing Staphylococcus aureus and inhibition by 2-aminoimidazole compounds
Kristen E. Livengood, Kevin L. Anderson, and Daina Zeng
North Carolina State University College of Veterinary Medicine, Raleigh, NC (Livengood, Anderson) and Agile Sciences,
Inc., Raleigh, NC (Zeng)
Staphylococcus aureus commonly causes chronic subclinical bovine mastitis, negatively impacting milk production and
quality of life for affected cows. Mastitis caused by S. aureus is difficult to cure. One factor that may reduce treatment success is the ability of S. aureus isolates to produce biofilm, a protective extracellular matrix of polysaccharide, protein, and
nucleic acid. Recently, 2-aminoimidazole (2-AI) compounds derived from the sea sponge product, ageliferin, were shown to
inhibit biofilms, representing new strategies for antibiotic therapy. In this study, we tested seven genotypes of mastitis-producing S. aureus and successfully demonstrated their ability to form biofilms in vitro. Additionally, we demonstrated the
ability of three 2-AI compounds to inhibit biofilm production. Biofilm production by diverse S. aureus genotypes (B1-B7)
was quantified using a crystal violet stain based biofilm assay. The most robust biofilm producer (B6) was tested in a biofilm
inhibition assay using three 2-AI compounds, AGL-348, AGL-396, and AGL-324, at concentrations from 100 mg/mL to
0.78 mg/mL. Percent of biofilm formation was determined using untreated control wells, and 50% inhibitory concentration
values (IC50s) were calculated. Biofilm production varied among genotypes, with B6 being the most robust biofilm producer. Biofilm inhibition varied among 2-AI compounds, with AGL-324 giving the lowest IC50 of 0.81 mg/mL. Biofilm
production of B6 may be a factor in its widely dispersed presence in mastitis in NC. The 2-AI compounds inhibited biofilm
production. Future steps include evaluation of AGL-324 in reducing antibiotic minimum inhibitory concentration values
against antibiotic-resistant S. aureus mastitis isolates.
Student Support: North Carolina State University CVM and Fund for Discovery
Evaluation of methods to preserve field samples for detecting elephant endotheliotropic herpesviruses
Courtney Loeschel and Paul Ling
College of Veterinary Medicine, Texas A&M University, College Station, TX (Loeschel); Department of Molecular
Virology and Microbiology, Baylor College of Medicine, Houston, TX (Loeschel and Ling)
Lethal elephant endotheliotropic herpesvirus (EEHV)-associated hemorrhagic disease in juvenile elephants has been documented worldwide, including in wild elephants. While EEHV has impacted captive elephants significantly in North America
and Europe, its impact on range country elephant populations remains unknown. To better understand the impact of this
disease, increased surveillance of samples from elephants in range countries for EEHV is needed. A barrier towards this
goal is the lack of adequate storage and temperature control in many locations in range countries, which may compromise
biological samples prior to arrival in a testing laboratory. The goal of this project was to evaluate methods to preserve DNA
in blood and mucosal samples in which the samples were subjected to potential conditions that might be encountered in
the field, particularly exposure to high temperatures. Samples of blood and mucosal swabs were spiked with EEHV1 viral DNA and incubated at 378C for three or seven days with Qiagen RNAprotect, Qiagen RNAlater, or no stabilizer, and
analyzed using an established EEHV qPCR assay. Surprisingly, we found that EEHV viral DNA in whole blood was very
stable without any need for a stabilizing agent, even when incubated at 378C for seven days. In contrast, preliminary data
indicates that mucosal samples at 378C might require stabilizing solutions. This project validated a protocol to detect EEHV
in whole blood samples subjected to conditions potentially experienced in the field. This will facilitate better detection and
characterization of EEHV in elephants in range countries.
Research Grant: A grant from Houston Zoo to Dr. Paul Ling
192
Induced GI tract microbial dysbiosis does not accelerate disease progression in SIV-infected Asian macaques
Mackenzie E. Long, Alexandra M. Ortiz, Miriam Quinones, and Jason M. Brenchley
National Institutes of Health, National Institutes of Allergy and Infectious Diseases, Laboratory of Parasitic Diseases,
Bethesda, MD
Disruption of the intestinal epithelium during acute HIV infection results in the translocation of microbial products from
the intestine to other areas of the body. These microbial products then stimulate the immune system and exacerbate disease
progression. Recent data have suggested that microbial translocation is associated with alteration of the composition of the
GI tract microbiome in HIV-infected individuals. However, this microbial dysbiosis is not observed in SIV-infected rhesus
macaques and the dysbiosis observed among HIV-infected individuals is possibly attributed to risk factors for HIV and not
the virus infection itself. To empirically assess the hypothesis that microbial dysbiosis contributes to disease progression via
increasing systemic immune activation, a vancomycin treatment protocol was established to induce sustained dysbiosis in a
non-human primate model for HIV infection. To ensure the subjects of the study exhibit sustained dysbiosis using the vancomycin protocol, 16S sequencing is used to identify fecal microbial members in samples collected at different time points
prior to and post SIV infection for each subject of the study. Despite induction of significant GI tract microbial dysbiosis,
we observed that animals treated with vancomycin had similar SIV viral loads, CD4 T cell loss, and inflammation compared
to control animals. Moreover, the animals with GI tract microbial dysbiosis progressed to simian AIDS at a similar rate to
control animals. Results from this study suggest that microbial dysbiosis does not significantly contribute to SIV disease
progression.
Research Grant: Funding for this study was provided in part by the Division of Intramural Research/NIAID/NIH. The
content of this publication does not necessarily reflect the views or policies of DHHS
Student Support: CBSTP Summer Internship Program in Biomedical Research for Veterinary Medical Students
Effect of dietary antioxidant deficiency on canine central vision: a pilot study
Ariana P. Lopez, Melanie L. Foster, Korinn E. Saker, Jessica Harris, Danielle Myzk, Christine M. McGahan,
Freya M. Mowat
Department of Clinical Sciences, (Lopez, Foster, Mowat), Department of Molecular Biomedical Sciences (Saker, Harris,
McGahan), Department of Population Health and Pathobiology, (Myzk), College of Veterinary Medicine, North Carolina
State University, Raleigh, NC
Oxidative damage to the retina is an important component of the prevalent blinding human disease, age-related macular degeneration (AMD). This disease targets the cone-dense central region of the retina known as the macula, but the exact mechanisms of the cone damage have not been established. Aspects of macular susceptibility cannot be modeled accurately in
rodents, as they do not possess a region where cones are concentrated. Canines, however, have a well-characterized central
cone-rich region called the area centralis. We sought in this pilot study to examine the clinical effects of oxidative damage
to this area of the retina in five normal beagle dogs using an established model of dietary antioxidant (vitamin E) deficiency.
Every two weeks, the retinal function was evaluated using electroretinography and the testing of visual navigation ability;
retinal morphology was examined using retinal photography and optical coherence tomography. Baseline examinations
were performed, and then three subjects underwent dietary deficiency in the antioxidant vitamin E through their diet, while
two subjects remained on a diet containing normal levels of vitamin E. After six weeks of deficiency, no significant differences in visual function or retinal morphology could be identified between dogs fed the vitamin E deficient diet and dogs fed
the control diet. The plasma alpha-tocopherol assay showed only mild deficiency in dogs fed the vitamin E deficient diet. A
diet modification to further reduce levels of vitamin E is currently underway, in addition to planning ocular administration
of an oxidant to accelerate disease.
Research Grant: NCSU Comparative Medicine Institute, Saker Nutrition Research Fund
Student Support: NIH T35 Interdisciplinary Biomedical Research Training Program Grant
193
Characterization of vascular remodeling in the canine brain following Traumatic Brain Injury
Melissa Lopez, Amanda D. Hazy, Michelle H. Theus, Bernard S. Jortner
Tuskegee School of Veterinary Medicine, Tuskegee, AL (Lopez), Department Biomedical Sciences and Pathobiology,
Virginia-Maryland Regional College of Veterinary Medicine, Blacksburg, VA (Hazy, Theus, Jortner).
Every year in the U.S., traumatic brain injury (TBI) accounts for 1.7 million visits to emergency rooms and of those patients
who survive, they add to the 5.3 million Americans (2% of the population) who are left with permanent long-term cognitive
and motor disabilities (1). Research into neuroprotection, regeneration, and repair following TBI can be applied to various
age categories and fields such as: combat related head trauma, children who acquire trauma while on the playground, the
elderly who suffer from a fall and sports related trauma. Stimulation of new blood vessel growth and remodeling, neovascularization, has gained recent attention in the brain due to its inextricable link to neuronal recovery. A better understanding
of the mechanism(s) that help support and repair the damaged cerebrovascular network, will have a tremendous impact on
functional outcome after TBI. The current study aimed to characterize microvascular and arteriole remodeling in the canine
brain using necropsied samples of a 2-year old boxer at 10 days post-TBI. Tissue samples from the (results pending) region
showed (results pending). Further studies investigating the gene expression changes in these vessels will help improve our
understanding of the molecular underpinnings controlling the vascular response following TBI.
b-cell glucagon-like peptide-1 receptor improves islet morphology after vertical sleeve gastrectomy in mice
Jon Lou, Darline Garibay, Anne K. McGavigan, Seon A. Lee, James V. Ficorilli, Amy L. Cox, M. Dodson Michael,
Kyle W. Sloop, Bethany P. Cummings
Department of Biomedical Sciences (Lou, Garibay, McGavigan, Lee, Cummings), College of Veterinary Medicine,
Cornell University, Ithaca, New York, USA and Endocrine Discovery (Ficorilli, Cox, Michael, Sloop), Lilly Research
Laboratories, Eli Lilly and Company, Indianapolis, Indiana, USA
Vertical sleeve gastrectomy (VSG), a surgical obesity treatment, results in type 2 diabetes (T2DM) remission days after
surgery prior to significant weight loss. However, the mechanisms by which this occurs have not been established. VSG
increases postprandial glucagon-like peptide-1 (GLP-1) secretion. This incretin hormone stimulates insulin production, insulin secretion, and b-cell protection. Surprisingly, studies using whole body GLP-1R knockout mouse models or GLP-1R
antagonists suggest that GLP-1R signaling does not contribute to glucose regulation after VSG. Recently, our lab confirmed
that GLP-1R signaling contributes to improved glucose regulation after VSG using a b-cell specific tamoxifen-inducible
knockout mouse model, eliminating potential compensatory pathway development that may have confounded previous
studies. We hypothesized that b-cell GLP-1R does this by improving islet morphology and insulin production. Male 2
month old inducible b-cell specific Glp-1r+/+ (WT) and Glp-1r-/- (KO) littermates were placed on a high fat diet (HFD) for
1.5 months, then switched to tamoxifen supplemented HFD for the rest of the study. Four month old mice underwent VSG
or sham surgery. One and a half months after surgery, tissues were collected. Paraffin embedded sections (n=3) were fluorescently stained for insulin and quantified. VSG WT mice had decreased b-cell area per islet compared with WT sham-operated (S WT) mice. However, b-cell area per islet did not differ between S KO and VSG KO (avg. b-cell area per islet: S
WT = 56486190, VSG WT = 35786501, S KO = 496561194, VSG KO = 589561927 mm2; P<0.05 S WT vs VSG WT).
This demonstrates for the first time that VSG improves islet morphology through b-cell GLP-1R signaling.
Research Grant: Cornell University, SUNY Graduate Diversity Fellowship, Cornell Comparative Cancer Biology Training
Program, President’s Council of Cornell Women, Eli Lilly and Company, and R21CA195002-01A1
Student Support: NIH Training Grant 5T35OD010941-07
194
Samantha Lovering, Alex Molesan, and Kathleen Kelly
Department of Biomedical Sciences, Cornell University College of Veterinary Medicine, Ithaca, NY
Myocarditis is considered rare in dogs, but its occurrence is associated with devastating effects, including death or permanent cardiac damage. While the etiology is often unknown, systemic viral infections or bacterial agents are often implicated.
The northeastern United States is a Lyme disease endemic area with high rates of Borrelia burgdorferi infection in dogs.
Clinical disease associated with B. burgdorferi infection of dogs is infrequent and generally vague with clinical signs including fever, arthritis, anorexia, lymphadenopathy, glomerulonephritis, and, rarely, myocarditis. Given the high incidence of B.
burgdorferi infection in dogs, we hypothesized that Lyme carditis is under-recognized in canine patients and may account
for myocarditis of unknown etiology. For our study, DNA was extracted from archived formalin-fixed paraffin-embedded
tissues from 28 myocarditis cases identified from the New York State Animal Health Diagnostic Center archives. Controls
were matched based on age. Conventional polymerase chain reaction, gel electrophoresis, and sequencing were used to
detect B. burgdorferi DNA in heart samples. Tissues were examined histologically and scored for myocardial necrosis, inflammation, and fibrosis by a blinded pathologist. Identifying etiologies common to canine myocarditis will have important
clinical benefit and may direct empirical treatments for suspected myocarditis. Please see poster for results.
Student Support: NIH Training Grant 5T35OD010941-07
Sudden acquired retinal degeneration syndrome in Western Canada: a retrospective (2002-2016)
Danica R. Lucyshyn, Bianca S. Bauer, Stephanie C. Osinchuk, Bruce H. Grahn, and Lynne S. Sandmeyer
Department of Small Animal Clinical Sciences, Western College of Veterinary Medicine, University of Saskatchewan,
Saskatoon, SK
Sudden Acquired Retinal Degeneration Syndrome (SARDS) is a leading cause of sudden, permanent blindness in dogs.
Little is known about the etiology or pathogenesis and no treatment exists. SARDS is reported to affect overweight, spayed,
middle-aged, female, small and mixed breed dogs. Dogs often present with signs of hyperadrenocorticism (HAC). This
study investigated signalment, clinical manifestations and laboratory results of dogs diagnosed with SARDS at the Western College of Veterinary Medicine (WCVM). Medical records search identified SARDS dogs presenting to the WCVM
from 2002-2016. Age, breed, sex, weight, ophthalmic examination, concurrent signs of HAC and laboratory results were
obtained. The population (n=91) was 47% spayed and 5% intact females, 41% castrated and 7% intact males. Mean age was
8.18 +/- 2.50 years. Purebreds (65%, n=59) were more affected than mixed-breeds (35%, n=32); Dachshunds (17%), Miniature Schnauzers (20%), and Pugs (15%) were overrepresented. Most (67%, 39/58) weighed under 11.3 kg. Systemic signs
of HAC were seen in 75% (36/48); 6 were tested but only 2 diagnosed with HAC. Ophthalmic examination showed 59%
(24/43) had reduced pupillary light reflexes and 50% (45/91) had mild retinal degeneration. Blindness was most often present for 30 days (24%, 19/78) before diagnosis. The most common abnormal laboratory test was the chemistry panel (95%,
20/21) with increased liver enzymes (71%, 15/21). Proteinuria was present in 77% (13/17). Results support the reported
predisposition of middle-aged, small breed dogs with signs of HAC but contradict reported high incidence in females. Low
concurrent diagnosis of HAC and SARDS exists. Many patients show ophthalmoscopic changes at time of diagnosis.
195
Evaluating human pluripotent stem cell-based photoreceptor transplantation in rodent models
Allison L. Ludwig, Joe Phillips, Patrick Barney, Christopher Lee, Jee Min, Shivani Jain, David M. Gamm
School of Veterinary Medicine (Ludwig), Waisman Center (Phillips, Barney, Lee, Min, Jain, Gamm), McPherson Eye
Research Institute (Ludwig, Phillips, Gamm), and Department of Ophthalmology and Visual Sciences (Gamm),
University of Wisconsin, Madison, WI
Vision loss due to photoreceptor (PR) dysfunction and death is a leading cause of blindness worldwide. Despite advances
in human pluripotent stem cell (hPSC) therapies, no PR replacement treatments currently exist for human patients as the
safety and efficacy of such studies must first be tested in animal models. Using existing rodent models, the ongoing aims of
our study are to optimize hPSC-derived PR production, refine cell delivery techniques, evaluate visual function, and investigate cell survival and migration. To track PR production in hPSC cultures, a PR specific reporter line was created. Using
protocols developed in the Gamm lab, hPSC-PRs were generated and differentiated to day 80-90 for peak PR precursor cell
production. hPSC-PR production was confirmed via PR-specific antibody immunostaining, including CRX, RECOVERIN,
RXRG, and S-OPSIN. Cells were then dissociated for transplantation and animals were monitored for 2 weeks. Immunohistochemistry allowed for analysis of surgical technique, survival of differentiated PRs, and cellular activity after delivery.
Transplanted cells were identified histologically with antibodies specific for reporter expression, human nuclei, and PR
cytoplasm, showing subretinal PR survival at 2 weeks. Electroretinography and optokinetic tracking were performed to establish baseline retinal electrical function and visual acuity for different lines. Several lines were tested, including immune
compromised Foxn1-RNU rats and J/NU mice to test safety and cell survival as well as s334ter rats and rd10 mice to test
efficacy. Ongoing studies will focus on determining the time course for transplanted hPSC-PR survival, integration, and
potential functional rescue in retinal disease models.
Research Grant: Foundation Fighting Blindness, Retina Research Foundation, McPherson Eye Research Institute, Reeves
Foundation, NIH P30 HD03352, Choroideremia Research Foundation
Latasha Ludwig, Rebecca Egan, Monica Baquero, and Brandon L. Plattner
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Canada
Mucosal surfaces, including those of the gastrointestinal and skin, are the frontlines of host defense against a wide variety
of infectious agents and pathogens. It is at these surfaces where initial host defenses either fail or are overcome by invading
pathogens in order for infectious disease to become established in any host. Our laboratory is working toward understanding the role of innate lymphocytes, especially gamma-delta T cells, during early host-pathogen interactions, and how they
contribute to host defense. It is known that these cells are among the earliest of all cells recruited to infection sites in several
species and that they preferentially localize to mucosal surfaces. We recently showed that distinct subsets of gamma-delta
T lymphocytes are present in the epithelium lining the gastrointestinal tract of young calves. The purpose of this study is to
determine the distribution and character of innate T lymphocytes throughout the gastrointestinal tract and skin in calves as it
is hypothesized that these cells are central to successful defense of mucosal surfaces. A multispectral fluorescence imaging
approach was used on tissues from the gastrointestinal tract and skin of healthy calves and our data show that the highest
proportion of gamma-delta T lymphocytes are present in the distal intestine and oropharynx. Distribution of innate T cells
in the distal intestine was further investigated using flow cytometry. These locations have an abundance of lymphoid tissue
(Peyers patches), and are hypothesized to be common primary sites of pathogen entry. This data will be a baseline for future
work to compare lymphocyte recruitment in these tissues after infection.
Research Grant: OMAFRA, NSERC
Student Support: Andrea Leger Dunbar Summer Research Assistantship, Dairy Farmers of Ontario
196
Matthew Lundquist, Hiruni Harischandra, Michael Kimber
Department of Biomedical Sciences, College of Veterinary Medicine, Iowa State University, Ames, Iowa
Parasitic nematodes pose a significant medical and veterinary global health concern. Lymphatic Filariasis (LF) is a Neglected Tropical Disease caused by filarial nematode worms including Brugia malayi. Over 120 million people are afflicted with
LF and 40 million people have the debilitating and social stigmatizing clinical form known as elephantiasis. The interaction
of host and parasite during LF is delicately balanced but poorly understood. Here we explore a novel mechanism by which
B. malayi modulate the host immune response with RNA and proteins delivered by extracellular vesicles called exosomes.
Macrophages are key mediators of the early immune response to infection. We examined the modulatory effect on cultured
macrophages by exosome-like vesicles (ELVs) collected from different life stages of B. malayi. By looking at macrophages
treated with these ELVs, we can see that different life stages of B. malayi elicit a differing response in macrophages. Macrophages were treated with purified ELVs and their responses were measured using ELISA, Griess assay and qPCR. Increased
IL-6 production was observed in the ELISA assay by macrophages treated with ELVs secreted by L3 stage parasites. High
levels of nitric oxide production were observed in the Griess assay by L3 treated macrophages compared with low or nonexistent levels in macrophages treated with other life stages. Supporting this observation, there was an upregulation of inducible nitric oxide synthase 2, demonstrated by qPCR. These data describe stage-specific classical activation of macrophages
in contrast to alternative activation as may have been expected and support a novel mechanism of immunomodulation by
filarial nematode parasites.
Research Grant: NIH/NIAID R21 AI117204
Student Support: NIH/NIAID R21 AI117204
Profile of gap junction intercellular communication in canine mammary carcinoma
Savannah Luu and Annelise Nguyen
Department of Diagnostic Medicine/Pathobiology, College of Veterinary Medicine, Kansas State University,
Manhattan, KS
Incidence of mammary carcinoma in dogs is three times that of humans. Mammary tumors are the most common type of
cancer in intact female dogs and account for about half of all neoplasms in these dogs. Previous studies showed that cancerous cells have a loss of cell-cell communication. The goal of this study is to characterize cell-cell communication in canine
mammary carcinoma. Three cell lines - CMT12, CMT27, and CF41.Mg - derived from canine mammary tumors, were used
to profile the cell communication pathway, consisting of connexin proteins and the protein kinase C (PKC) family. Seven
connexins and three PKC antibodies were used in immunofluorescence and Western Blot analyses. The results show a clear
differential pattern of connexins in these three cell lines. A significant decrease of connexin 43 in CMT27 cells compared
to connexins 26, 32, 36, 45, 46, and 50 was observed in both approaches of immunofluorescence and western blot analyses.
Previous data demonstrated that CMT27 cells have a relatively high rate of proliferation, suggesting that the loss of connexin 43 may contribute to cancer formation. Further analysis into the mechanism of the loss of connexin 43 in CMT27 cells
was investigated and results show that PKC beta translocates from the cytosol to the membrane, suggesting that PKC beta
may impact the downregulation of connexin 43. Overall, the study provides an insight into the role of cell-cell communication in canine mammary carcinoma and potential for a therapeutic target.
Research Grant: Johnson Center for Basic Cancer Research
Student Support: NIH-T35OD010979
197
Soil-borne protozoans enable persistence of facultative intracellular pathogens in the environment
Tatyana Lyakhova, David Markman
Department of Microbiology, Colorado State University, Fort Collins, CO
Yersinia pestis, the etiological agent of plague is a natural pathogen of prairie dogs with spillover potential into human and
domestic animal populations. Although prairie dogs are a keystone species in many ecosystems, they are no longer a major
contributor to biodiversity due to mass culling campaigns attempting to reduce sylvatic plague frequency. Understanding
infection pathways and the environmental persistence of Y. pestis will enable prediction of outbreaks in prairie dogs, thus
contributing to conservation of prairie dogs and public safety. Although, it is well known that fleas serve as vectors for
plague, the source of infection for the origin-cases remain unclear, with current evidence suggesting the existence of a
cryptic environmental reservoir. Previous studies report that free-living amoeba (FLA) can be infected by Legionella pneumophila, which can further multiply within the protozoa. Furthermore, several other bacteria from the Yersinia genus are
known to resist amoebal digestion. FLA have two life stages, an active trophozoite stage and an inactive, environmentally
stable cyst stage. This study determines the ability of various facultative intracellular bacteria to survive inside FLA cysts
for prolonged periods of time. Thus, FLA may act as reservoirs of bacterial pathogens in soil over inter-epizootic timespans.
Part of this research assesses the amoebal and bacterial diversity of plague-endemic soils in North-eastern Colorado to identify candidate species for which intra-amoeba persistence may occur. This research will allow specific targeting of disease
emergence hotspots and FLA species susceptible to bacterial infection in order to control future outbreaks.
Research Grant: T35OD015130
Student Support: NIH T35 Fellowship Trainee Grant
Immediate treatment of hyperketonemia ameliorates negative impacts on milk production but not comorbidity risk
Tyler N. Mack, Rafael C. Oliveira, Ryan S. Pralle, Garrett R. Oetzel, Heather M. White
Department of Medical Sciences, University of Wisconsin-Madison School of Veterinary Medicine, Madison, WI (Mack,
Oetzel); Department of Dairy Science, University of Wisconsin-Madison, Madison, WI (Oliveira, Pralle, White)
Hyperketonemia (HYK) is one of the costliest diseases facing the dairy industry, affecting 40 to 60% of cows (primarily
in the first three weeks post-calving). It is a metabolic response to negative energy balance that leads to excessive mobilization of fat and buildup of ketones in the bloodstream. The objectives of this study were 1) to examine milk production,
risk for displaced abomasum (DA) and herd removal for cows with HYK; and 2) to evaluate milk fat:protein ratio (FPR) as
a diagnostic tool for HYK. A convenience sample of 1,291 cows from two dairy farms located in southern Wisconsin was
enrolled. Blood b-hydroxybutyrate (BHB) was measured from each cow four times between 3 and 18 days in milk. Cows
with blood BHB concentrations $1.2mM were classified as HYK and treated immediately with oral propylene glycol, a
standard ketosis treatment. Mean blood BHB was 1.79 vs. 0.76mM for HYK vs. non-HYK cows (P<0.01). Milk samples
were collected at monthly intervals post-calving; milk production was greater for cows with HYK at the first nine monthly
milk tests(P<0.05). Cows with HYK were at greater risk for DA (9% vs. 2%, P<0.01) and removal from the herd (33% vs.
25%, P<0.01). Milk FPR $1.4 has been postulated to indicate HYK. However, 66% of HYK and 47% of non-HYK cows
had FPR $1.4; therefore, FPR is not adequate for diagnosing HYK. Early detection and prompt treatment of HYK ameliorates the drop in milk yield that typically follows ketosis; however, cows with HYK still remain at high risk for other adverse outcomes with negative economic implications. Early detection and prompt treatment of HYK is not sufficient; dairy
producers must emphasize prevention of HYK through management and nutritional strategies.
Research Grant: USDA Critical Agricultural Research and Extension (CARE) Grant, NIFA #2015-67028-23572
Student Support: School of Veterinary Medicine, University of Wisconsin-Madison
198
NSAIDs inhibit wound healing in intestinal and reproductive epithelial cell lines
Kelsey Madden, Bruce Schultz and Jim Lillich
Department of Anatomy and Physiology, College of Veterinary Medicine, Kansas State University, Manhattan, KS
Non-steroidal anti-inflammatory drugs (NSAIDs) inhibit migration of IEC-6 cells (epithelioid cell line derived from rat
duodenum) following mechanical wounding, but model systems to assess recovery of transepithelial electrical resistance
(Rte), a more sensitive measure of epithelial restitution, have not been reported. Additionally, work has not been conducted
to determine whether reductions in prostaglandin synthase (PGHS) activity could account for these observations. IEC-6
cells along with PVD9902 cells, derived from pig vas deferens, were employed. Cells were grown on solid substrates for
migration assays and treated with NSAIDs (ibuprofen, celebrex, SC-560, indomethacin) or vehicle for 48 (PVD9902) or 72
h (IEC-6). Monolayers were wounded and migration was measured 4 and 6 h later, respectively. PVD9902 cells were also
cultured on permeable supports for 2 wks. After confirming that 24 h NSAID exposure did not affect Rte, monolayers were
wounded and their resistance assessed every 6 h for 3 d. Prostaglandins (PGs) were quantified in mucosal and serosal media
before and 24 h after NSAID treatment. Each NSAID caused a significant reduction in migration in both cell types and a
12-18 h lag in Rte recovery. PGs were depressed 50-90% with NSAID treatment in both compartments. While decreased
PG concentrations correlated with reduced migration and delayed Rte recovery, it is not clear if this change accounts for
the diminished wound healing. Interestingly, mucosal concentrations of PGs were far greater than serosal concentrations,
suggesting that cells preferentially transport PGs to the lumen. This observation may help explain clinical manifestations of
NSAID toxicity which might have been previously overlooked.
Research Grant: 1 R15 DK091791-01
Student Support: (NIH)T35OD010979
Comparison of lymphocyte count and proviral load in beef and dairy cattle infected with bovine leukemia virus
Jacqueline E. Maeroff, Oscar J. Benitez Rojas, Vickie J. Ruggiero, Bo Norby, Rebecca LaDronka,
Paul Bartlett and Dan Grooms
Department of Large Animal Clinical Sciences (Maeroff, Benitez Rojas, Norby, Bartlett, Grooms) and the Comparative
Medicine and Integrative Biology Program (Ruggiero, LaDronka), College of Veterinary Medicine, Michigan State
University, East Lansing, MI.
Bovine leukemia virus (BLV) is a retrovirus of cattle responsible for enzootic bovine leukosis, which causes leukemia and/
or lymphosarcoma in some animals and can result in significant economic losses for producers. Cattle with BLV antibodies
were identified by milk or serum ELISA (AntelBio, East Lansing, MI). Blood samples from these cows were evaluated for
lymphocyte count (LC) by using QScout technology (Advanced Animal Diagnostics, Morrisville, NC) and proviral load
(PVL) using BLV-CoCoMo-qPCR (RIKEN, Wako, Japan). We found that higher lymphocyte counts moderately correlate
with higher PVL in both beef and dairy cattle with R2 values of 0.44 and 0.52 respectively. Optical density from the ELISA
test was poorly correlated to both PVL and LC in dairy and beef cattle with R2 values less than 0.23. LC and PVL means
were numerically higher in dairy (7607.405 lymphocytes/uL, 37447.64 copies/uL) than in beef cattle (7496.356 lymphocytes/uL, 35531.87 copies/uL). On farms with high BLV prevalence, it’s unrealistic to cull all BLV positive cattle. Cattle
with higher PVL are considered to be more infectious to their herdmates than low PVL cattle. Therefore, culling of cattle
with higher proviral loads is preferred. The current test for proviral load, which consists of qPCR, can be costly and time
consuming. Running qPCR tests on all ELISA positive cows is also unrealistic. This correlation between PVL and LC could
serve as a useful tool in identifying the most infectious cows with less cost and time required. We conclude that there is a
moderate, positive correlation between PVL and LC, and therefore LC could serve as a surrogate test to identify cows that
are at the highest risk of transmitting BLV to the rest of their herd.
Research Grant: United States Department of Agriculture National Institute of Food and Agriculture Award #2015-6702823652 and 2014-68004-21881
199
Investigation of sarcolipin as a cause of recurrent exertional rhabdomyolysis in horses
Kaitlin Maier, Shea Anderson, Keri Gardner, Carrie Finno, Stephanie Valberg
University of Minnesota (Anderson), Michigan State University (Maier, Gardner, Valberg)
Recurrent exertional rhabdomyolysis (RER) affects 5-7% of racing Thoroughbreds, Standardbreds and potentially Quarter
Horses and is characterized by cramping and muscle damage after moderate exercise. Dysregulation of sarcoplasmic reticulum intramuscular calcium cycling has been proposed to cause RER based on skeletal muscle contracture testing where
exposure of muscle to halothane or caffeine in a bath produced a lower contracture threshold in RER vs. normal horses. RER
is suggested to be inherited, but no genetic markers have been identified associated with RER and linkage studies have not
implicated calcium regulatory genes including RYR1 [calcium channel], CACNA1S [transverse tubule voltage sensor], and
ATP2A1 [SERCA1 ATPase]. We theorize that calcium dysregulation in RER horses may be caused by mutations in genes
encoding newly discovered proteins such as sarcolipin (SLN) that modify the activity of sarcoplasmic reticulum calcium ATPase (SERCA). Sanger sequencing of SLN was performed on DNA isolated from blood or muscle tissue of horses selected
from a repository of RER cases (6 Quarter Horses, 5 Standardbred horses, and 6 Thoroughbred horses) and 2 Thoroughbred
controls. Whole genome sequencing data from 31 horses of various breeds was also available as a control dataset. SLN
sequence was aligned with the reference genome and variants were compared between affected and control horses. Seven
intergenic variants were identified with two variants significantly associated with RER by x2 analysis. These two variants
were located 700-1000bp downstream of the SLN stop codon. As gene regulation is often influenced by variants in the 3’
untranslated region, these variants warrant further investigation in additional horses.
Research Grant: Morris Animal Foundation
Student Support: Merial Veterinary Scholars Program Grant and the MSU Graduate School
Radiographic and echocardiographic assessment of left atrial size in dogs with mitral regurgitation
Elizabeth L. Malcolm, Lance C. Visser, Kathryn L. Phillips, Lynelle R. Johnson
Department of Medicine & Epidemiology (Malcolm, Visser, Johnson), Department of Surgical & Radiological Sciences
(Phillips), School of Veterinary Medicine, University of California, Davis, Davis, CA.
Left atrial (LA) size is a powerful indicator of severity of myxomatous mitral valve disease (MMVD), the most common
heart disease in dogs. Thoracic radiography and echocardiography are two commonly used modalities to assess LA size.
However, determining LA size radiographically is primarily subjective and its comparison to echocardiography (clinical
gold standard) has not been well-studied. The objective of this study is to determine the feasibility and repeatability of an
objective measurement of LA size on radiographs called the vertebral left atrial size (VLAS), and to determine if VLAS
correlates with LA size determined echocardiographically. In this prospective observational study, LA size was determined
echocardiographically (indexed to the aorta [LA:Ao] in short and long axis) and radiographically via VLAS by a single,
blinded investigator in 103 dogs (88 with varying degrees of MMVD; 15 controls). VLAS was measured from a lateral thoracic radiograph by drawing a line from the carina to the caudal most border of the LA (where it summates with the caudal
vena cava) and indexed to the vertebral bodies. Preliminary analysis showed that VLAS exhibited a strong correlation to
LA:Ao in short (r = 0.71; P < 0.0001) and long (r = 0.76; P < 0.0001) axis. VLAS was feasible to measure in all dogs and
demonstrated to be a repeatable measurement when measured in right vs left lateral radiographs (average coefficient of variation 6.1%). Future work will determine the diagnostic accuracy of VLAS for diagnosing LA enlargement and cutoffs will
be compared to a radiologist’s assessment. Preliminary results suggest that VLAS may be a diagnostically valuable tool in
objectively assessing LA size on radiographs in dogs with MMVD.
Student Support: University of California, Davis, School of Veterinary Medicine Funds
200
Generation of Infectious Bronchitis Virus (IBV) M41 cDNA clones for use in chimeric IBV virus construction
Sara J. Mangosing, Greg Brennan, and Yvonne Drechsler
Infectious Bronchitis is a contagious upper-respiratory tract disease in birds that results in substantial losses in the poultry industry. It is caused by the avian gamma coronavirus, Infectious Bronchitis Virus (IBV), a 30-Kb, enveloped, positive-sense RNA virus that is transmitted through air or fecal shedding. It results in signs that include watery eyes, mucus in
the nares and trachea, coughing, nephritis, decreased egg production and quality, and decreased weight gain. It’s currently
unclear which IBV genes are important for pathogenesis; however, studies using chimeric IBV have shown that the replicase
gene complex, which had been thought to be strictly involved in viral replication, may contribute to pathogenesis through
interaction with host-cell pathways. In order to investigate regions involved in pathogenesis, such as the replicase genes, we
plan to generate chimeric IBV using vaccinia virus. This system will allow us to modify the full cDNA clone of avirulent
Beaudette-IBV with specific fragments of m41-IBV (respiratory pathogenic) by homologous recombination. As a first step,
I have specifically amplified four fragments of m41-IBV, using primers that were positioned to avoid instabilities, spanning
nucleotides 20420 to 27453 (structural genes). I cloned these fragments into Topo-XL and submitted them for Sanger sequencing. These fragments will then be cloned with a eukaryotic selection marker under control of VACV 7.5K vaccinia
promoter into a pUC19 vector via Gibson cloning. I have also started designing primers flanking the open reading frames
for non-structural proteins within the replicase complex. Upon successful generation of the chimeric viruses, infection will
be evaluated in ovo and in vivo.
Research Grant: Merial, Western University of Health Sciences College of Veterinary Medicine intramural funds
Student Support: Merial, Western University of Health Sciences College of Veterinary Medicine intramural funds
Characterization of N-linked glycan profiles in bat gastrointestinal tissues specific to influenza A viruses
Olivia Mann, Chun-Kai Yang, Scott Rush, Matthew E Grilliot, George Peng Wang, and Xiu-Feng Wan
Department of Basic Sciences (Mann, Yang, Grilliot, Wan), Mississippi State University College of Veterinary
Medicine, Mississippi State University, Mississippi State, MS; Department of Chemistry (Wang), Georgia State
University, Atlanta, GA
Bats are a potential reservoir for Influenza A virus (IAV), and it has recently been discovered that bats carry a newly-classified influenza-like virus. It has been demonstrated that this virus does not bind canonical human or avian receptors. The
central hypothesis is that bats harbor unique glycans to which this new influenza-like virus binds, and that bat-origin influenza A virus has unique receptor glycans distinct from human-origin influenza A viruses. To identify glycans in bat tissue,
sections of bat small intestines were harvested, processed, and subjected to hydrophilic interaction liquid chromatography
before being permethylated and identified via mass spectrometry. In addition, influenza-specific glycans were harvested
using Nickel-ionized HisTrap columns and purified recombinant H1 and H17 proteins. Glycan profiles of two different bat
species, Myotis austroriparius (My au) and Eptesicus fuscus (Ep fu), revealed that the majority of glycans resides in the
duodenum, with the former species having more total glycans than the latter. H1-specific glycans of the two species showed
that among the three portions of small intestine, the ileum contains more H1-specific glycan content, and most H1-specific
sialyation are alpha 2,6-linked. In addition, the H1-specific glycans from bat tissues are commonly fucosylated. Interestingly, My au had far more H1-specific glycans than did Ep fu by a ratio of approximately 4 to 1. Additional experiments are
still ongoing to identify H17 specific glycans, and validating the binding between the identified receptor and the viruses.
This project helps our understanding of natural history and tropisms of IAV, especially the newly-identified bat-origin IAV.
Research Grant: NIH Grant #T35OD010432 and NIH Grant R151R15AI107702-01
Student Support: NIH Grant #T35OD010432 and NIH Grant R151R15AI107702-01
201
Using behavior to establish a timed oxytocin protocol for estrous suppression in mares
Hannah Manning, and Erin Runcan
The Ohio State University CVM, Columbus, OH
Poor performance and undesirable behavior during estrus are common complaints made by horse owners and trainers.
Previous studies show an oxytocin injection protocol between days 7 and 14 of ovulation, reliably suppresses estrus in a
majority of mares (70%). However, the current protocol requires serial costly veterinary visits to determine the exact date of
ovulation. Thus, we hypothesize that by using behavioral indicators of estrus alone; we can use a timed injection protocol to
reliably suppress estrus without knowing the exact date of ovulation. All research was conducted at the Alice Lloyd Finley
Memorial Veterinary Research Farm utilizing 15 light-breed mares of various ages. The control group (n=4) was used to
confirm that the mares are cycling normally once every 21-28 days. Experimental mares (n=11) were determined to be in estrus by introducing them to a stallion and observing them for signs sexual receptivity. When such behaviors were observed,
this was considered Day 0 of the experimental protocol. Oxytocin injections were administered once daily from Day 8 to 17
following the initial signs of estrus. Serum progesterone levels were analyzed to evaluate for persistent elevation, suggesting
prolongation of the corpus luteum. Awaiting completion of the study, preliminary results show a trend of estrus suppression
in a majority of mares (6/11). Pending final results and statistical analysis, this study could create an inexpensive and more
practical way for clients to suppress estrus in performance mares.
Student Support: Groppe Research Fellow
Biomonitoring the effects of BPA and its metabolite BPAG during fetal life
Anujah Jenifer Mariampillai, Veronique Gayrard, Nicole Picard-Hagen, Catherine Viguie and Glenn Gauderat
Ontario Veterinary College, University of Guelph, Guelph, ON (Mariampillai). INRA, UMR 1331 TOXALIM-E3 GPE:
Gestation and Perturbation Endocrinienne, Ecole Nationale Veterinaire de Toulouse, Toulouse, France
(Gayrard, Picard-Hagen, Viguie, Gauderat).
Bisphenol A (BPA) is an endocrine-disrupting chemical widely used in polycarbonate plastic and epoxy resins production.
Its widespread human exposure is of great concern particularly during the sensitive period of fetal development. Previous
studies in our laboratory have shown BPA’s inactive metabolite, bisphenol A glucuronide (BPAG), to undergo deglucuronidation reactions back to free-BPA at the level of the fetal-placental unit. Thus maintaining a constant low level of basal fetal
BPA concentration. Our current study uses the in vivo pregnant sheep model to compare the effects of equimolar exposure
of BPA or BPAG on fetal tissue biomarkers. Two approaches were considered, a non-targeted proteomic approach to explore
proteomic shifts triggered by active BPA and a second approach to explore the targeted effects on insulinemia. Most of the
functions from the proteins altered were in relation to immunity and hormonal pathways. Fetuses treated with BPAG had
the ratio of mean plasma insulin concentrations before and after treatment, decrease with the dose administered. Glucose
stimulated insulin secretion was also decreased in the BPAG treated fetus compared to control. These finding suggest that
BPAG is not an inactive metabolite and should be considered when looking at BPA risk assessment in human exposure.
Research Grant: French Midi-Pyrenees, French National Research Agency, French National Center for Scientific
Research, UK Royal Society
202
Optimization of extraction and quantitation of cfDNA from equine plasma
Jacqueline L. Marinoff, Anna-Claire Bullington, and Maria S. Ferrer
Department of Large Animal Sciences, College of Veterinary Medicine, University of Georgia, Athens, GA
The aim of this study was to optimize a method to extract cell-free DNA (cfDNA) from equine plasma. A column-based
(QIA; QIAamp DNA Blood Midi Kit), magnetic (MAG; MagMAX cfDNA isolation kit), and phenol:chloroform:isoamyl
alcohol+glycogen (PCI) extraction method were compared. MAG was predicted to produce the best cfDNA yield, as it is designed to target small cfDNA fragments. Eight 2-mL samples of frozen equine plasma were processed by each method. The
products were quantitated using UV-Vis spectrophotometry (N; Nanodrop) and fluorometry (F; Qubit), and the sequence
of the housekeeping gene GAPDH was amplified using RT-PCR. PCI produced the highest cfDNA concentration (reported in ng/mL and mean6SEM) measured by N (PCI:79.568.7, MAG:3.460.3, QIA:3.760.3; P<0.0001; ANOVA) and F
(PCI:0.2960.06, MAG:0.00960.001, QIA:0.00760.0006; P=0.001; ANOVA). CtGAPDH was different among methods
(PCI:30.2860.32, MAG:33.0760.33, QIA:34.9660.58; P<0.001; ANOVA). There was also a difference in the frequency
of samples that yielded concentration below F’s detection limits (PCI: 0%, MAG:37.5%, QIA:62.5%; P=0.02; x2). QIA
was considered the poorest extraction method. So, 7 additional 2-mL plasma samples were processed using MAG and PCI.
cfDNA concentration was higher with PCI (N:131.57620.01, F:0.354 6 0.048) than MAG (N:2.4760.34, F:0.05760.023)
(P<0.001; T test). CtGAPDH was lower in samples extracted with PCI (29.8260.27) compared to MAG (31.5460.43;
P<0.001; T test). It was concluded that PCI provided the best results in terms of cfDNA yield and amplification of GAPDH.
These findings will be used to develop a RT-PCR-based assay to determine fetal sex through amplification of the SRY gene
from extracted fetal cfDNA in maternal plasma.
Research Grant: Theriogenology Foundation and College of Veterinary Medicine intramural funds
Breanna K. Marshall, Karla Marquez-Nogueras, and Silvia N J Moreno
University of Georgia College of Veterinary Medicine, Athens, GA (Marshall), Center for Tropical and Emerging Global
Diseases, University of Georgia, Athens, GA (Marquez-Nogueras, Moreno)
Toxoplasma gondii is an Apicomplexan parasite that infects one third of the world’s population. T. gondii is a zoonotic
parasite and can infect both animals and humans. The disease, toxoplasmosis, can be devastating in immunocompromised
patients and in the unborn fetus of pregnant women. Transmission occurs via the shedding of oocysts through the feces
of cats, which can infect a variety of intermediate hosts. Once inside the host, parasites transform into tachyzoites, which
replicate inside host cells resulting in their lysis, a process called lytic cycle, which consists of invasion, replication, egress
and finding of a new host cell. Previous studies have demonstrated that calcium enhances virulence traits in T. gondii, as it
regulates molecules that are essential for its lytic cycle. Nonetheless, most of the molecular players involved in these signaling pathway are unknown. We hypothesize that calcium binding proteins regulate important proteins involved in calcium
entry and calcium homeostasis. In order to understand this pathway, we characterized the role of a calcium-binding protein,
TgEFHDCP60. Calcium binding proteins are known for having EF-hand domains and for regulating other molecules/
enzymes. Using C-terminal tagging, we localized this protein in both extracellular and intracellular parasites. Using the
CRISPR-Cas9 system, we created knockouts of this protein, which was validated through Quantitative PCR. Plaque assays
revealed smaller plaques in comparison to the wild type cells suggesting a growth phenotype. In addition, using Fura-2AM
loaded parasites, we observed higher calcium entry than in wild type cells. Altogether, our data suggests that TgEFHDCP60
plays a role in the regulation of calcium entry.
Research Grant: NIH AI096A36, NIH AI110027
203
Adoption of recommended hand hygiene practices to protect public health at agricultural fairs
Alison R. Martin, Michele M. Zentkovich, Jaqueline M. Nolting, Dimitria A. Mathys, Dixie F. Mollenkopf,
Andrew S. Bowman
Department of Veterinary Preventive Medicine, College of Veterinary Medicine, The Ohio State University,
Columbus, OH
Agricultural fairs are a prime location for zoonotic disease transmission. Specifically, the transmission of influenza A virus
(IAV) from exhibition swine to humans occurs in this setting via direct and indirect contact. Public health organizations
have established hand hygiene guidelines for animal settings to prevent the spread of zoonotic disease. We hypothesized that
although these guidelines are well established, adoption of these recommendations by fair officials and/or fairgoers is low.
Over 3 years, 107 fairs were analyzed to determine presence and functionality of hand hygiene stations, as well as presence
of signage for human health risks and hand wash procedures. Overall, 87.9% of fairs had functional hand hygiene stations.
Interestingly, 9 fairs added hygiene stations the year after exhibition swine tested IAV positive. During 2016, presence of
IAV and bacteria on these stations, and their usage by fairgoers, was tested at 8 fairs. Up to 5 functional hand hygiene stations were tested per fair representing animal housing and human only use areas. Stations were wiped with an electrostatic
cleaning cloth, moistened with BHIB and taken to the lab for testing. All 35 samples were RRT-PCR negative for IAV and
are undergoing bacterial testing. For each fair, up to 2 of the tested stations were observed for an hour each to determine the
proportion of fairgoers using stations upon exiting an animal area. Only 9.1% of people used these stations. Overall, fairs
have followed hand hygiene guidelines by providing hygiene stations. However, the gap identified between these guidelines
and their adoption by fairgoers illustrates a strong need to educate the public on zoonotic disease transmission and prevention.
Research Grant: St. Jude Center of Excellence for Influenza Research and Surveillance
Aileen Martinez, Sidney I. Roberts, Joy E. Tomlinson, Jonathan Cheetham
Cornell University, CVM, Ithaca, NY
Over 300,000 Americans are affected by peripheral nerve injuries each year. Macrophages play a key role in repair after
injury. We are investigating how manipulation of macrophage phenotype affects functional recovery. Here we investigate
use of a rotarod technique, which has traditionally been used to evaluate central nervous system deficits, to evaluate recovery after peripheral nerve injury. We performed a pilot study to evaluate differences between fixed speed and accelerating
approaches and determined the appropriate amount of pre injury training. We then evaluated the effect of global deletion for
Interferon gamma receptor on recovery after crush injury. Mice were trained for 7 days, and then subjected to proximal or
distal sciatic crush or transection with ligation (negative control). Recovery was evaluated on post-operative days 5, 7, 10,
13, 18, 21, and 24 using an accelerating rotarod test. Overall, the rotarod may serve as useful test for longitudinal assessment
for functional recovery after nerve injury.
Research Grant: RO3 NIDCD Manipulation of macrophage phenotype in peripheral nerve injury
204
Evaluation of novel drug therapies against human breast and canine mammary carcinomas
Samantha Marie Martinez, Tasha Miller, Kim Tran, Michael Bittner, Heather Wilson-Robles
Small Animal Clinical Sciences Department, Texas A&M University, College Station, TX (Martinez, Miller, Tran,
Wilson-Robles); The Translational Genomics Research Institute (Bittner)
Carcinoma of the breast is the most common invasive cancer in women. This year alone over 240,000 women will be diagnosed with invasive breast cancer and over 40,000 women are expected to die from the disease. Dogs are the most commonly affected domestic species with a prevalence in intact female dogs being two times higher than women. MLN9708 (ixazomib) is a next-generation proteasome inhibitor, lapatinib is a small molecule inhibitor of Her2/neu (a growth factor receptor
commonly mutated in aggressive breast cancer) and SH-4-54 is a small molecule, nonphosphorylated STAT3 inhibitor. For
this study, MLN9708, lapatinib, and SH-4-54 were used against human and canine mammary carcinoma cell lines. Both canine and human cell lines were shown to have significant decreases in viability when treated with MLN9708 and lapatinib.
However, SH-4-54 did not significantly affect viability of any of the cell lines. Forty-eight hour apoptosis assays supported
this finding demonstrating that MLN9708 and lapatinib were more effective at cell killing than SH-4-54. Downstream gene
targets of the drugs were analyzed using quantitative real time PCR. The human cell lines had more predictable responses
with up regulation of pro-apoptotic factors and down regulation of survival factors. The canine cell lines had more variable
responses suggesting an altered mechanism for response. Future studies will focus on evaluating these drugs in combination
and further elucidating the mechanisms of these drugs in vitro and in vivo.
Research Grant: The Fred and Vola Palmer Chair in Comparative Oncology
Native and recombinant PD-1, PD-L1, and PD-L2 variants for study of porcine circovirus associated disease
Caitlin Mason, William R. Huckle, and Tanya LeRoith
Department of Biomedical Sciences and Pathobiology, Virginia-Maryland College of Veterinary Medicine,
Blacksburg, VA
Co-infection of porcine circovirus type 2 and other pathogens, such as porcine reproductive and respiratory syndrome virus,
can result in porcine circovirus associated disease (PCVAD), which leads to high rates of mortality among swine. Viral-induced lymphocyte depletion is the hallmark of systemic PCVAD. One mechanism by which this depletion may occur is
through upregulation of programmed death ligands (PD-L1 and PD-L2) on the surface of dendritic cells. These ligands
bind to programmed cell death protein 1 (PD-1), a receptor present on T cells, promoting T cell anergy and apoptosis. With
upregulation of the ligand, the frequency of binding increases and leads to lowered immune defenses. We hypothesize that
truncated variants of PD-1 and its ligands, either those occurring naturally or provided as recombinant agents, may modulate
PD-L1/PD-1 mediated dendritic cell/T cell interactions in PCVAD. To test this hypothesis, we sought evidence of native
pre-mRNA processing variants using end-point PCR and 3’-RACE analysis. Results to date suggest that RNA splice variants of PD-L1 and PD-L2 that omit key extracellular domains occur in the normal porcine lymph node. Characterization
of these variants, including assessment of their expression patterns and functional integrity, is ongoing. In parallel with this
screen for endogenous variants, we have prepared synthetic cDNAs encoding truncated forms of PD-1, PD-L1, and PD-L2
predicted to be secreted from cells. These recombinant forms, proteins that lack transmembrane domains but still capable of
recognizing their cognate binding partners, will be useful in further investigation of porcine dendritic cell/T cell interaction
in the normal immune response as well as in PCVAD.
Research Grant: NIH 4T35OD011887-10
205
Effects of anti-inflammatory glucocorticoids on hemodynamics and cardiac function in clinically healthy dogs
Allison K Masters, Darren J Berger, Johann F Coetzee, Wendy A Ware, Jessica L Ward
Department of Veterinary Clinical Sciences (Masters, Berger, Ware, Ward) and Department of Veterinary Diagnostic and
Production Animal Medicine (Coetzee), College of Veterinary Medicine, Iowa State University, Ames, Iowa
The use of glucocorticoids is presently limited in patients with heart disease due to the concern for possible precipitation of
congestive heart failure. The objective of this study was to investigate previously proposed mechanisms by which glucocorticoids could cause progression of heart disease, including fluid retention from mineralocorticoid effect, transient hyperglycemia causing intravascular fluid shift, increased afterload due to systemic hypertension, and direct cardiac remodeling. In
this prospective clinical trial, clinically healthy dogs with allergic dermatitis (n = 11) were given anti-inflammatory doses of
oral prednisone (1mg/kg/day) for 14 days, followed by a taper and washout period. Hemodynamic, biochemical, and echocardiographic variables were measured prior to prednisone administration (d0) and at days 7, 14, and post-washout (d35).
We hypothesized that prednisone administration would cause no changes in any measured variables. To test this, paired
two sample t-tests were used to compare variables between baseline and d7, d14, and d35. Results showed no significant
changes from baseline in blood glucose or sodium/potassium ratio. Significant increases from baseline did occur in blood
pressure at d7 (p = 0.0061), normalized IVSd at d35 (p = 0.012), and myocardial relaxation at d7 (p = 0.012). Overall, this
study demonstrates that anti-inflammatory doses of glucocorticoids in clinically healthy dogs have the potential to impact
cardiac structure and function. Further data analysis for other measured variables and investigation using matched control
dogs is currently underway.
Research Grant: ISU CVM Seed Grant
Student Support: Boehringer Ingelheim Vetmedica
Fate of exosporium proteins BclA, BclB, and BxpB in germinating Bacillus anthracis spores
Michael J. Mattia, Hsin-Yeh Hsieh, George C. Stewart
MU Veterinary Research Scholars Program (Mattia) and Department of Veterinary Pathobiology and Bond Life Sciences
Center (Hsieh, Stewart), University of Missouri, Columbia, MO, USA
Bacillis anthracis is a spore-forming, Gram-positive rod that is widely distributed throughout the world and responsible for
zoonotic disease in mammals, especially ruminants. When the environment lacks sufficient resources, the bacteria undergo
sporulation and enter a metabolically inert state, becoming surrounded by a hardy spore coat. B. anthracis spores also possess an additional outer layer, the exosporium, consisting of a basal layer and a hairlike glycoprotein nap layer. Exosporium
proteins include BclA (in the nap layer), BclB (thought to reside beneath the nap layer), and BxpB (in the basal layer). The
exosporium contains a “cap” region (accounting for about 25% of the exosporium) and “non-cap” region (the remaining
75%). When external conditions are again suitable to sustain growth, the cell will undergo germination and emerge from the
spore via the opening of the cap. Precisely how the cap opens to allow exit of the bacterium from the spore is not understood.
BclB is found exclusively in the non-cap region. It has been demonstrated that mutants lacking BclB form spores with a
defective cap, suggesting an essential role of BclB in cap formation. We hypothesize that BclB may be a target of proteases,
serving to open the cap after activation and outgrowth of the bacterium. To test this, we harvested spores and stimulated
germination in a nutrient-rich medium while inhibiting cell growth with antibiotics. We extracted exosporium proteins by
boiling in the presence of detergent, size-fractionated the extracts via SDS-PAGE, and identified BclA, BclB, and BxpB proteins with Western blotting with antiserum specific for these proteins. The amount and size of the proteins were compared
to those of ungerminated spores.
Research Grant: Grant R21AI112725 from the National Institute of Allergy and Infectious Diseases.
Student Support: An endowment established by IDEXX-BioResearch.
206
Antigen presenting cells promote protective immunity in catfish exposed to live Edwardsiella ictaluri vaccines
Matty May, Adef Kordon, Hossam Abdelhamed, Hamada Ahmed, Joo Youn Park, Attila Karsi, and Lesya Pinchuk
Department of Basic Science, College of Veterinary Medicine, Mississippi State University, Starkville, Mississippi
We have identified the presence of Langerhans-like cells (LCs) containing Birbeck granules in the gill of uninfected catfish
by immunohistochemistry. Also, we showed that catfish peritoneal macrophages are capable of actively phagocytosing and
killing E. ictaluri opsonized with sera from catfish vaccinated with live attenuated vaccine (LAV) candidates compared
to control fish. These results lay a foundation for our present study that is to determine the vaccine-induced production of
innate and adaptive immune responses through catfish antigen presenting cells, such as dendritic cells, macrophages, and B
cells. In this study, we demonstrate that in vivo E. ictaluri infection dramatically changes the numbers of LCs from visually
undetectable in uninfected fish to highly abundant numbers of LCs in the infected catfish gill. Furthermore using a qPCR
approach, we found several principle differences in the CD8a T cell co-receptor gene expression levels in both anterior
kidney and spleen between the vaccinated and non-vaccinated groups suggesting the development of T cell-mediated immunity. Our in vivo infection study also showed a significant correlation in colonization of vaccine strains, active antigen
uptake, and bacteria killing properties of anterior kidney mononuclear cells. We expect that these findings will be important
in understanding the mechanisms of innate and adaptive immune responses against the most effective vaccines.
Research Grant: United States Department of Agriculture
Student Support: National Institutes of Health T35OD010432
Immunophenotype comparison of equine bone marrow-derived and synovial membrane-derived
George T. McClung, Renata Linardi, Kyla F. Ortved
Department of Clinical Studies at the University of Pennsylvania School of Veterinary Medicine New Bolton Center,
Kennett Square, PA
In the equine patient, osteoarthritis (OA) can lead to severe disabilities, and is both highly prevalent and very costly to
manage. Injury to the hyaline cartilage is a key instigating cause of OA, which is a progressive disease with no definitive
cure. Several studies have investigated the use of mesenchymal stem cells derived from bone marrow (BM-MSCs) to
enhance cartilage repair; however, full chondrogenesis remains a significant challenge. Synovium-derived mesenchymal
stem cells (SD-MSCs) may have superior chondrogenic capabilities allowing them to be a more effective cell source for
cartilage repair. The broad objective of this experiment was to compare the immunophenotype of BM-MSCs and SD-MSCs
from healthy and osteoarthritic joints. We hypothesized that these cell types would have similar immunophenotypes. Bone
marrow was obtained from the sternabrae of and synovial membrane was obtained from healthy and osteoarthritic joints of
healthy horses. MSCs were culture expanded to passage 2 prior to immunophenotyping via flow cytometry. Cells were
stained with antibodies against CD29, CD44, CD90, CD105, CD45RB, CD79alpha, MHCI, and MHCII. Flow cytometry
was used for quantification of fluorescence. Comparison of immunophenotypes will allow characterization of SD-MSCs
and direct further quantification of cultured cell populations. Future aims of this project will investigate the growth rates and
chondrogenic potential of these cell types.
Research Grant: Raymond Firestone Trust Research Grant
Student Support: NIH grant T35-OD-010919 and Merial
207
Characterization of tertiary lymphoid organs in a natural model of Multiple Sclerosis
Megan McGeehan, Priscilla Farias, Miles Miller, Melissa Sanchez, Jorge Ivan Alvarez
Department of Pathobiology, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, PA
Multiple sclerosis (MS) is an idiopathic demyelinating disease characterized by leukocyte infiltration across the central
nervous system (CNS) vasculature leading to the formation of demyelinating lesions and major neurological impairment.
Granulomatous meningoencephalomyelitis (GME) is an idiopathic demyelinating disease in dogs that closely resembles
pathological features of MS. In MS, immune cell infiltrates resembling tertiary lymphoid organs (TLOs) have been correlated with accelerated clinical disease. TLOs are highly organized lymphoid aggregates arising from the interaction of distinct
immune cell subsets and resident stromal/endothelial cells at sites of chronic inflammation. They are architecturally and
functionally similar to secondary lymphoid organs and support the differentiation of lymphocytes. It is unclear how TLO
genesis occurs in the CNS so to better understand their development, we characterized immune cell infiltration in 40 GME
cases. GME is characterized by lymphohistiocytic infiltration in the white matter and cortex localized predominantly in the
perivascular spaces. The subarachnoid space is more heavily infiltrated by lymphocytic populations bias towards B cells.
These infiltrates highly resemble lymphoid follicles and the cells within are organized along a conduit system. Our ongoing
histo-immunological analysis is determining the prevalence of various cell types composing meningeal TLO-like structures.
To date, we have detected immune and stromal cells and factors known to support TLO development. This study demonstrates the presence of TLO-like structures in the subarachnoid space of GME cases and is the first documenting TLO-like
structures in chronically inflamed CNS.
Student Support: NIH/Merial Veterinary Scholars Program
Morphology of peritoneal endometriotic lesions using an in-vivo mouse model of retrograde menstruation
Victoria McLean, Quanxi Li, Romana Nowak
Department of Animal Sciences (McLean, Nowak) and Department of Comparative Biosciences (Li),
University of Illinois at Urbana-Champaign, Urbana, IL
Endometriosis is a painful, chronic disease in which endometrial tissue grows outside of the uterus in women of childbearing age. The condition commonly causes pelvic pain, dysmenorrhea, and dyspareunia, while also resulting in infertility
in 30-50% of cases. The theory of retrograde menstruation is the most commonly accepted mechanism of endometriosis,
stating that blood and endometrial fragments flow backward through the oviduct and exit through the ovarian bursa into the
peritoneal cavity. The purpose of this preliminary experiment is to investigate the morphology of peritoneal lesions using
a mouse model of endometriosis that mimics the process of retrograde menstruation found in humans. Endometrial tissues
from female mice primed with pregnant mare serum gonadotropin (PMSG) were transplanted into the peritoneal cavities
of ovariectomized, syngeneic mice treated with estradiol (E2) at regular intervals. Immunohistochemistry (IHC) was performed on lesion sections using antibodies against epithelial marker claudin-4, proliferation marker Ki67, endothelial marker CD31, smooth muscle cell marker ASMA, and mesothelial marker calretinin. Previous studies reveal an abundance of
proliferating stromal cells, myofibroblasts, and mature blood vessels within these sections, which this study also confirms,
but it is unknown what processes are occurring in the mesothelial cells that are in contact with the endometrial cells. If the
results confirm the disappearance of mesothelial cells, further studies will determine whether these cells are undergoing a
mesothelial-to-mesenchymal-transition (MMT), causing them to lose their adhesive properties and secrete factors that may
aid in the establishment of endometriotic lesions.
Research Grant: NIH 1R21 ES026388
Student Support: NIH T35 OD011145
208
Obesity promotes breast tumor development and metastases in an immunocompetent mouse model
Rachel Q. McMahon, Lauren E. Hillers, and Lisa M. Arendt
Department of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin, Madison, WI
Obesity has been established as a significant risk factor associated with the development of breast cancer in postmenopausal
women. Furthermore, obesity is correlated with larger, higher grade breast tumors and increased incidence of lymph node
metastasis. Under conditions of obesity, visceral fat is associated with chronic inflammation and infiltration of macrophages. Recent studies in our laboratory suggest that macrophages recruited to an obesity-like microenvironment promote
tumor growth. Based on this observation, we hypothesized that obesity will accelerate tumor growth and promote increased
frequency of metastasis to the lungs in obese mice, potentially with increased recruitment of macrophages. Using an immortalized breast cancer cell line, EO771 cells, we directly examined breast tumor development and metastasis using a high
fat diet model of obesity. EO771 cells were implanted into the inguinal mammary glands of obese and lean C57Bl/6 female
mice. Obese mice demonstrated decreased latency to tumor formation along with significantly larger tumor volumes at end
stage compared to lean mice. Using histologic examination, lung metastases were identified at a significantly greater frequency in obese mice than lean mice. Using immunohistochemistry, we evaluated the recruitment of alveolar macrophages
and reactive fibroblasts within lung tissue using markers CD44 and smooth muscle actin (SMA). No significant differences
were detected in the number of alveolar macrophages, however, we detected SMA+ fibroblasts surrounding metastases. Our
findings support the hypothesis that obesity promotes breast tumor growth and metastasis, but the mechanism by which this
occurs remains unclear.
Research Grant: Susan G. Komen CCR15332611
Scott McMullin, Sammira Rais-Rohani, Bryn Brazile, and Jun Liao
Mississippi State University CVM (McMullin), Starkville, MS Department of Biological Engineering
(Rais-Rohani, Brazile, Liao), Mississippi State University, Starkville, MS
In space, astronauts experience several physical abnormalities due to microgravity. Some of these include intraorbital and
intracranial pressure changes similar to the effects of idiopathic intracranial hypertension. When exposed to microgravity,
many astronauts experience an enlarging of the optic nerve, resulting in the buckling of the optic nerve sheath. These abnormalities may result in blurred vision, which can ultimately lead to permanent blindness. It is believed that the optic nerve
may be an integral part of both the cause and the solution to preventing or treating this affliction. Although there have been
several studies concerning the correlation between intracranial pressure and the optic nerve, to the best of our knowledge
there is no study that investigates the biomechanical properties of the optical nerve itself. Porcine heads were obtained from
a local abattoir, and the optic nerves were dissected. The optic nerves were trimmed of all excess material leaving the sheath
intact and placed in phosphate buffered saline (PBS). Compression tests were performed on the optic nerves by mounting
the nerves onto a Test Resource compression machine and wetted with PBS solution to obtain the tissue buckling behavior.
The compression test shows that the optic nerve shows an increasing linear relationship between strain and stress until the
stress reaches ~15Pa, at which point it plateaus from strains roughly 0.22 to 0.26. The stress then shows a sharp exponential
increase after this until the end of the test. This study established a strong basis for optical nerve biomechanics. Future work
will include histology, bending tests, and computational simulation of the effects of abnormal pressures.
Research Grant: unknown
209
McSweeny I, Weltman J, and Baker T
University of Wisconsin-Madison
Sleep apnea (SA) is characterized by recurrent reductions (hypopnea) or absence (apnea) of breathing during sleep. ~70% of
insulin resistant individuals (IR) exhibit SA. While it has been proposed that coincident IR and SA are linked primarily by
body composition, recent work in rats showed development of SA with IR induction regardless of body composition. Since
chronic exposure to intermittent hypoxia (CIH) also increases the incidence of apnea and hypopnea during sleep in rats and
humans, IR may lead to a self-perpetuating phenomenon wherein IR leads to periodic disruptions in breathing during sleep
causing cyclical exposures to hypoxia which beget further hypopnea/apnea. The purpose of this study is to test the hypothesis that coincident dietary induced obesity (DIO) and CIH increase hypopneas and apneas in the mouse. BL6 mice were
randomized to normal chow or high fat chow (12 wks, n=4, respectively) for 6 wks prior to baseline plethysmography. Subsequently all mice underwent CIH (90 sec, 6.5% O2, 90 sec, 21% O2, 12 hrs, 14 days) and plethysmography was performed
12 hrs following the final exposure. Data were analyzed for tidal volume and respiratory, apnea and hypopnea frequency.
Preliminary results suggest an increase in hypopnea following CIH (12.5/hour +/- 24 DIO, 19.7/hour +/- 10.8 normal chow)
with no effect of diet. We speculate that while CIH leads to destabilized breathing in mice as it does in rats and humans, the
addition of high fat diet for 6 weeks does not further destabilize breathing. Additional studies are necessary to confirm these
findings. It is possible that a longer duration of DIO and/or DIO induced by high sugar diet may be necessary to reveal the
impact of IR on breathing during sleep.
Research Grant: NIH R01-HL10551
Student Support: NIH T35OD011078 and NIH T32-OD010423-09
Sur1 and TRPM4 expression in the spinal cord of dogs with progressive myelomalacia
Julia E. Medland, Baoyan An, Barbara Sherry, and Natasha J. Olby
Progressive myelomalacia (PMM) is a rare, fatal condition in dogs occurring secondary to spinal cord injury (SCI). Necrosis
and hemorrhage extend from the lesion epicenter along the spinal cord. In rodent models of SCI, the ion channel TRPM4Sur1 is upregulated in the vasculature around the lesion, resulting in a massive influx of calcium into the cell, inducing cell
death and destruction of the local microvasculature. We hypothesized that TRPM4-Sur1 upregulation results in progressive
endothelial cell death in PMM. Study aims were to establish level of expression of TRPM4-Sur1 in normal canine spinal
cords and to compare this to their level of expression in spinal cord from dogs with PMM. Spinal cord segments from three
dogs with diagnosed PMM and four control dogs were collected at the time of euthanasia for RNA extraction. qPCR was
performed to quantify expression of TRPM4 and SUR1, VWF (endothelial cells), and RBFOX3 (neurons), with GAPDH.
TRMP4 and Sur1 were expressed in the normal canine spinal cord. In affected dogs, TRPM4 and Sur1 expression was low
in the thoracic and lumbar cord around the lesion epicenter, but increased with distance from lesion, with the highest expression at the level of C2. Neuronal RBFOX3, but not VWF, mirrored this expression pattern. Level of expression of Sur1
at C2 was significantly higher than controls (p=0.017). Our data suggest SUR1 is upregulated in neurons surrounding the
injury site, causing progressive necrosis. Following verification of this finding, controlling the expression of this channel in
canine SCI may improve outcomes.
Student Support: NIH T35 Interdisciplinary Biomedical Research Training Program (IBRTP)
210
Pathophysiologic environmental factors diminished efficacy of ceftiofur and penicillin with gentamicin
Ashlee C. Melhado, Theresa Beachler, Jennine Lection, Scott Bailey
Veterinary Student (Melhado, Lection), Department of Clinical Sciences (Beachler, Bailey), College of Veterinary
Medicine, North Carolina State University, Raleigh, NC
Ceftiofur is frequently used to treat equine reproductive diseases. Efficacy has been established in vitroand drug penetration
to the uterus has been confirmed in non-pregnant animals. However, little is known about the influence of pathophysiologic
environmental factors on drug efficacy. The objective of this study was to investigate the effect of different fluid environments on ceftiofur (CEF), in comparison with penicillin and gentamicin (PG). We hypothesized that either CEF or PG would
have a bactericidal effect on Escherichia coli(EC) and Streptococcus equi zooepidemicus (SZ) in Mueller Hinton Broth
(MHB), equine lochia (PPF), and purulent fluid obtained from a mare with pyometra (PYF). A modified time kill trial was
used to determine bacterial concentration at 8 and 24 hours (h) after inoculation in each fluid, with and without antibiotics
added based on published pharmacokinetic data. Antibiotics were considered bactericidal or effective when they resulted
in $3-fold decrease in colony forming units, per the CLSI guidelines. CEF was effective against EC in MHB at 8h, but not
at 24h. Against SZ, CEF was ineffective by 8h, but effective at 24h. CEF was ineffective in PPF and PYF against EC at 8
and 24h. CEF was ineffective in PPF and PYF against SZ at 8h and bactericidal at 24h. PG was bactericidal in PPF against
EC and SZ at 8 and 24h. PG was ineffective in PYF against EC at either 8 or 24h and against SZ at 8h, but bactericidal at
24h. The efficacy of both antibiotics was significantly impacted by fluid type. PG, but not CEF maintained broad spectrum
bactericidal efficacy in PPF. These findings suggest that CEF may not be the best treatment choice for postpartum mares at
risk for uterine infections.
Student Support: Veterinary Scholars Program at NCSU CVM
Investigating the role of Ndr kinase in retina development.
Rebecca Meltzer, Eliot Smith, Frank Luca
Retinal development is a complex and highly coordinated process that involves the interplay of multiple signaling networks
that govern cellular development, function, spatial organization, and establishment of intricate neuronal connections between
many different cell types. Mutations that disrupt retinal development commonly lead to retinal degeneration and blindness.
Recently, a mutation in the canine Ndr2 gene was shown to cause early retinal degeneration (ERD), as characterized by
increased cell proliferation, apoptosis, and progressive retinal degeneration. Despite the discovery of Ndr2’s involvement in
ERD, the mechanisms of the kinase in retinal development and function are unknown. Because Ndr2 and its paralog Ndr1
negatively regulate the transcriptional co-activator YAP in small intestinal epithelial cells via phosphorylation, and Yap is
also known to influence retina development, we hypothesize that Ndr1/2 kinases regulate retinal cell proliferation via YAP.
We will test this hypothesis by determining the relationship between Ndr kinases and YAP in retinal cell proliferation and
apoptosis in zebrafish. If this hypothesis is correct, then Ndr gene deletions should cause increased cell proliferation and
apoptosis and a corresponding increase in YAP mediated transcription. Thus, we will examine retinal cell proliferation and
apoptosis in Ndr+/+ and Ndr-/- zebrafish retinas using immunohistochemistry and qPCR of cell cycle markers. We will also
quantify differences in YAP-mediated gene transcription in wild type and Ndr-/- mutant fish. Resolving the relationship
between Ndr, YAP, and retinal development may lead to the development of therapies to induce photoreceptor regeneration
and prevent degeneration.
Research Grant: NIH R01 GM097327
Student Support: NIH T35 Training Grant T35OD010919 and Merial
211
Oviduct morphology and Prolactin receptor expression in N. American River Otters and Asian Small Clawed Otters
Rachel Melvin and Dalen Agnew
Michigan State CVM, East Lansing, MI
Otter populations worldwide have been declining. Gaps in knowledge of their reproduction have slowed captive breeding
efforts. The North American River Otter (NARO), Lontra Canadensis, and the Asian Small-Clawed Otter (ASCO), Aonyx
cinerea, are well represented in captivity and have two distinct reproductive cycles. The NARO is monoestrus and seasonal
with a marked embryonic diapause, whereas the ASCO is polyestrus and non-seasonal and lack diapause. There is little
information on the role of the oviduct in delayed implantation. I hypothesized that NARO and ASCO will differ in oviduct
length and histomorphology, as well as prolactin receptor expression in the ovary and uterus. Oviducts from both species
were dissected from archived tracts held by the Reproductive the Reproductive Health Surveillance Program. The oviduct
length was measured and cross-sectional segments from the proximal, middle, and distal third of its length were trimmed,
embedded, sectioned, stained with H&E, and the histomorphology of each will be analyzed (ImagePro Plus ). Slides were
labeled with antibodies for prolactin (PRL) receptor and the density, intensity, and location were analyzed. We found the
NARO oviducts are significantly longer than the ASCO oviducts. Histomorphology is pending; however the ratio of non-ciliated:ciliated cells along the oviduct epithelium of NARO oviducts is expected to be greater than ASCO. PRL receptor
antibodies successfully labeled otter reproductive tissues; however, preliminary findings found no significant difference in
PRL receptor expression. Additional knowledge of their reproductive tract and in particular, the physiology of diapause will
allow improved reproductive interventions in these threatened species.
Research Grant: AAZV, AZA Reproductive Management Center
Student Support: Merial/Michigan State University College of Veterinary Medicine and Graduate School Fellowship
A single mutation in the vaccinia virus RNA polymerase inhibits multiple host restriction factors
Gerardo Mendoza Jr, Stefan Rothenburg, Greg Brennan
College of Veterinary Medicine (Mendoza, Brennan) Western University of Health Sciences, Pomona, CA. Division of
Biology (Rothenburg), Kansas State University, Manhattan, KS
About 70% of diseases that have recently emerged within the human population, such as HIV/AIDS, Ebola, and Zika virus,
have a zoonotic origin. To infect new hosts, viruses must overcome host restriction factors such as protein kinase R (PKR)
and the oligoadenylate synthetase (OAS)/Ribonuclease L (RNase L) pathway. Both PKR and RNase L are activated by intracellular dsRNA, a pathogen associated molecular pattern (PAMP) produced during many viral infections. PKR prevents
viral replication by inhibiting translation and the OAS/RNase L pathway degrades both cellular and viral RNA leading to
cellular apoptosis. The Brennan lab has identified a mutation in the vaccinia virus RNA polymerase, A24R, that inhibits
PKR (A24R*). Similarly, a different mutation in A24R (IBTR90) has been shown to inhibit RNase L by reducing the accumulation of dsRNA during replication. The A24R* mutation does not appear to inhibit PKR through the same mechanism;
therefore, it is unclear whether A24R* can also inhibit RNase L. To investigate this question, I performed RNase L degradation assays on infected cells. In this assay, cells with activated OAS/RNase L will degrade ribosomal RNA in a stereotypical
pattern. I infected cells with VC2, A24R*, IBTR90, A35R* and DE3L at an MOI of 3.0, then isolated total RNA 24 hours
post infection. I found that A24R* infected cells have an inhibitory effect on RNase L comparable to that of IBTR90. Our
data suggests that these two independent A24R mutations inhibit RNase L through two distinct mechanisms. Furthermore,
mutations in the RNA polymerase complex may be a mechanism to inhibit multiple host restriction factors.
Research Grant: Merial Veterinary Scholars Program; Western University of Health Sciences Intramural Funding
Student Support: Merial Veterinary Scholars Program; Western University of Health Sciences Intramural Funding
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Domains of CD163 involved in infection of Type I and Type II PRRSV
Stephen Mercer, Ana Stoian, Bob Rowland
Department of Diagnostic Medicine and Pathobiology, College of Veterinary Medicine, Kansas State University,
Manhattan, KS
Porcine Reproductive and Respiratory Syndrome has been shown to be one of the most economically destructive diseases
affecting swine worldwide. In the U.S. alone, the most recent estimates indicate the disease caused $664 million in losses
from the national breeding and growing herd annually from 2005-2010. CD163, present on macrophages, has been shown
to play a critical role in infection as a putative receptor for both PRRSV and African Swine Fever Virus. Physiologically,
CD163 functions as a scavenger receptor for the hemoglobin-haptoglobin complex. CD163 is composed of nine scavenger
receptor cysteine-rich domains. Deletion mutants are being produced that will allow determination of which domains are
important to viral infection by Type I & II PRRSV. In previous work, a number of constructs were created including domains
2-end, 3-end, 5-end, 7-end, and 9-end. These constructs were used to transfect the non-permissive cell-line HEK293T. The
cells were then challenged with Type II PRRSV and levels of infection were measured. Currently, these constructs are being
used to transfect HEK293T cells which will be infected with Type I PRRSV to determine if the same domains are important
to both Type I and Type II infection. New constructs are also in production including domains 1-2, 1-5, 1-7, 1-9, and 6-end.
Constructs are being produced by using PCR to amplify specific regions of the CD163 gene. The DNA fragments obtained
are cloned into a TOPO vector and transformed into competent cells. A double restriction enzyme digestion is performed to
remove the fragments from the TOPO vector. The fragments are then ligated into pcDNA3.1 which is used to transfect the
permissive cells.
Research Grant: Kansas NBAF Transition Funds
Student Support: Center of Excellence for Emerging and Zoonotic Animal Diseases
Monitoring bull behavior to determine characteristics of bull dominance and the role in social ranking
Micaela L. Mertens, David K. Hardin, and Richard F. Randle
College of Veterinary Medicine, Iowa State University, Ames, IA (Mertens), Department of Veterinary and Biomedical
Sciences, College of Veterinary Medicine, University of Nebraska - Lincoln, Lincoln, NE (Hardin, Randle)
Natural mating accounts for more than 90% of pregnancies in beef cattle and the male is an important factor. Factors that
influence the ability for a bull to achieve paternity include semen quality, scrotal circumference, mating ability, and social
dominance. The first three qualities are measurable, but social dominance is not easily determined, yet becomes an important factor when multiple bulls are used in breeding situations. Social dominance is defined as the social ranking that
develops among bulls. The purpose of these studies is to observe bull behavior to better determine criteria for ranking bulls
based on their social dominance order. In study one, 5 synchronized yearling heifers were placed with three Angus bulls,
15 to 20 months old. The bulls were individually marked and behaviors were recorded for two days. In the second study,
79 adult cows, previously synchronized and timed bred were placed with two 15 month old Angus bulls and one 4 year old
Angus cross bull. Observations were recorded for seven days. Observed behaviors of the bulls included following females,
sniffing vulvas, mounting and aggressiveness toward other bulls. At times females in the sexually active group appeared to
court a bull by licking and mounting the bull. In study one, 1 of the 3 bulls spent more time with the sexually active females
on day 1 while on day 2, one of the other bulls assumed this role. In study two, the 4 year old bull spent the most time with
the sexually active females. Based on our observations, we conclude that the dominance order was not fully established in
the yearling bulls in study one while the dominance order in study two appeared to be established and stable with the older
bull appearing to be dominant.
Research Grant: Merial
Student Support: Merial and Iowa State University College of Veterinary Medicine
213
Sarah Middlebrooks, Jeffrey Eells, Shirley Guo-Ross, Ciarra Smith
Mississippi State University CVM, Starkville, MS
Schizophrenia is a poorly understood disease that alters the CNS by eliciting positive symptoms and cognitive symptoms;
however, most of the etiopathology is unknown resulting in inadequate management strategies. Toxoplasma gondii is an
intracellular protozoan that infects one third of the population worldwide. Many studies have shown that T. gondii is an
important risk factor for schizophrenia via its ability to encyst in the brain and alter neurotransmitter levels. Another risk
factor is the mutation of the Nurr1 nuclear receptor that is needed for dopamine synthesis and dopamine neuron survival. To
investigate the mechanisms through which a genetic mutation and infection with T. gondii can contribute to risk for schizophrenia and cognitive deficits, we determined the behavioral effects of T. gondii infection in adult mice. Furthermore, we
incorporated the Nurr1-null heterozygous (+/-) mice into this study to observe any cumulative effects that could contribute
to cognitive deficits. Our hypothesis was that the +/- mice will be more sensitive to T. gondii infection which will elicit more
severe cognitive deficits. Adult mice were tested for open field activity and spatial learning using a Barnes maze. Both genotype and treatment effects were observed in which Nurr1-null +/- and T. gondii infected mice exhibited longer total escape
latency times in the Barnes maze. In an open field test, Nurr1-null +/- mice were more active. Another important effect was
that both T. gondii treated groups experienced weight loss. By combining these factors into one model, we were able to
observe cognitive deficits through quantitatively scoring our mice models based on their behavior.
Research Grant: Funding provided by National Institutes of Health 5T35OD010432
Student Support: National Institutes of Health
Effect of a supplement containing curcumin on lameness due to osteoarthritis or degenerative disease
Travis Miller, Laura Riggs, Mandi Lopez, Michael Keowen, Kelsey Koenig, Catherine Takawira, Frank Andrews
Equine Health Studies Program, Department of Veterinary Clinical Sciences(Miller, Riggs, Keowen, Koenig, Andrews),
Laboratory for Equine and Comparative Orthopedic Research (Lopez, Takawira), Louisiana State University, School of
Veterinary Medicine, Baton Rouge, LA
Lameness is an abnormal stance or gait caused by either a structural or a functional disorder of the locomotor system which
results in the inability to move normally. Recently, studies have suggested that curcumin may be effective against the
pain and inflammation associated with osteoarthritis and rheumatoid arthritis in people. The purpose of this project is to
determine the clinical efficacy of an oral herbal supplement containing curcumin (Longvida) for improvement of lameness
in horses due to osteoarthritis. Two healthy thoroughbreds horses from the Equine Health Studies Program research herd
were identified with front limb lameness, localized to the forelimb (fetlock, carpus) by a board certified equine surgeon.
Radiographs of the affected joint confirmed osteoarthritis. Horses were studied in a crossover design, and housed in stalls
for the duration of the study. On day 0, force plate analysis as well as lameness exams were performed. During phase 1,
starting on Day 1, one horse received the supplement (Curcumin, 2 ounces top-dressed on grain, once daily), and the other
horse was the control and received grain alone. Force plate analysis and lameness exams were performed again on day 15,
and will be performed again on day 30. Changes in impulse due to variations in stance time were noted at days 0 and 15.
An improvement in lameness is expected by day 30. Blood samples were collected through IV catheters on days 0 and 14,
and centrifuged at 3000 rpm for 15 minutes. The plasma was separated and stored in cryogenic vials at -80C, and will be
used for pharmacokinetic evaluation. Phase 2 will begin following a 2 week washout period, and the treatment and control
will be switched.
Research Grant: Equine Health Studies Program
Student Support: Merial Summer Scholars Program, LSU School of Veterinary Medicine
214
Zachary G. Millman, Kelly L. Hughes, Anne C. Avery
Department of Microbiology, Immunology and Pathology, College of Veterinary Medicine and Biomedical Sciences,
Colorado State University, Fort Collins, CO.
T-zone lymphoma (TZL) is an indolent, non-effacing subtype of T-cell lymphoma characterized by the loss of expression
of CD45, unique cytological and histological features, and in some cases signs of immunosuppression. Due to their rarity,
T-cell lymphomas are poorly studied in human patients compared with B-cell lymphomas. TZL is common in dogs and has
a strong breed predilection, making it a useful model for studying the origins and function of one type of T-cell lymphoma.
Previous gene expression profiling performed in our laboratory using NanoString technology, implicated the Th2 subtype
of T helper cells as the cell of origin for TZL. This gene expression data also revealed significant expression of the proteins
galectin-1 and galectin-3, which can drive T helper cell differentiation toward a Th2 phenotype. Based on this data, we hypothesize that TZL should express high levels of the Th2 transcription factor GATA-3, and low levels of the Th1 transcription factor, T-bet. We also suspect that the loss of expression of the CD45 antigen may cause upregulation of galectins 1 and
3, which normally bind CD45 on the cell surface to induce apoptosis. Using canine antibodies for GATA-3, T-bet, CD45,
galectin-1, and galectin-3, we seek to validate the gene expression studies and establish Th2 cells as the cell of origin for
canine TZL via western blots, immunohistochemistry, and flow cytometry.
Research Grant: None.
Utility of ATP-bioluminescence measured by a luminometer to monitor hygiene of milk replacer for dairy calves
Kelly Mills, Fiona Maunsell, G. Art Donovan
University of Florida CVM, Gainesville, FL
Bioluminescence uses the enzyme luciferase which reacts with ATP produced by living organisms and creates a photon of
light, which is then measured. ATP-bioluminescence has been used to assess bacterial contamination of surfaces and liquids.
Studies have demonstrated reliability of ATP-bioluminescence for detecting bacteria in raw and ultra-heat treated milk. Recently, hand held portable ATP-bioluminescence units (luminometers) that can read ATP activity in less than a minute have
been marketed. The luminometer could offer significant advantages over standard bacterial culture as results are available
immediately. In calf milk replacer (MR) feeding systems, bacterial contamination of >105 cfu/ml is associated with reduced
health. The goal of this project was to investigate the hypothesis that a luminometer (SystemSure Plus , Hygeina, LLC,
Camarillo, CA) would measure calf MR systems hygiene rapidly and reliably. Water and MR inoculated with various levels
of bacteria were tested, and free and total ATP readings were compared with bacterial culture results. The luminometer was
unable to detect bacteria in undiluted MR, likely due to high levels of background interference, so samples were diluted
10-fold in water. The luminometer was able to detect $107 colony forming units (cfu/ml) of bacteria in the diluted MR,
with a sensitivity of 0.83 and a specificity of 0.93 using a cutoff of 300 relative light units of free ATP. In spiked water, the
luminometer was able to detect bacteria at <104 cfu/ml. These findings indicate that for MR, there is reduced ability to detect
bacterial contamination compared with water. Use of the hand-held luminometer for determination of MR system hygiene
is not recommended.
Research Grant: University of Florida Foundation- Dr. Maunsell
Student Support: 2016 Merial Summers Scholars Program
215
STAT3 activation via the IL6/gp130 pathway in sepsis-related and hyperinsulinemic laminitis
Melanie Mironovich, Mauria Watts, Kathryn Dern, and James K. Belknap
Department of Clinical Sciences, The Ohio State University College of Veterinary Medicine, Columbus OH
Laminar failure in equine laminitis is characterized by cytoskeletal and adhesion protein dysregulation on laminar basal
epithelial cells, leading to displacement of the distal phalanx and severe lameness. Similarly, these are the first two steps
in epithelial cells undergoing transformation to tumor cells (epithelial to mesenchymal transition/EMT). Specifically, these
early events in epithelial tumorigenesis are reported to occur due to aberrant activation of STAT3 via the IL-6 family of cytokines and the IL6-R/gp130 complex. Due to similarities between cellular events in laminar and intestinal epithelial cells, we
investigated the roles of IL-6/IL6-R and gp130 in STAT3 activation, a key player in the progression from an inflammatory
state to cancer observed in inflammatory bowel disease. On the basis of these observations, we hypothesized that laminar
failure in laminitis is due to gp130/STAT3 activation induced by elevated levels of IL-6 and IL6-R. Analysis of archived
samples from both equine metabolic syndrome (EMS) laminitis, replicated using the eugylcemic hyperinsulinemic clamp
(EHC) model and sepsis-related laminitis (SRL), simulated using the oligofructose overload (OF) model revealed up to a
2,000 fold increase in IL-6 concentrations, along with marked phosphorylation of STAT3 in both the OF and EHC models
of laminitis. IL6-R was found to be increased in the EHC model, and has not yet been evaluated in the OF model. We will
continue to investigate IL6-R and gp130 levels. The data obtained suggests IL-6 activation of gp130/STAT3 signaling may
play a significant role in the development of laminitis, and needs to be further studied to determine its specific roles and
targets for pharmaceutical intervention.
Research Grant: Grayson Jockey Club Research Foundation, USDA
Antibiotic alternatives for the reduction of morbidity and mortality in veal calves
Elspeth Mitchell, Jessica Pempek, Katy Proudfoot, Mary Masterson, Jeff Lakritz, and Greg Habing
Department of Veterinary Preventative Medicine (Proudfoot, Masterson, Habing) and Department of Veterinary Clinical
Sciences (Lakritz), College of Veterinary Medicine, The Ohio State University, Columbus, OH
Diarrhea is a top cause of morbidity and mortality in veal and dairy calves. Calves are fed milk replacers containing antibiotics to reduce the severity and incidence of illness. This study tests the efficacy of antibiotic alternatives in preventing
diarrhea in veal calves. Lactoferrin, an iron-binding protein found in milk, has been shown to increase calf performance.
Cinnamaldehyde, an essential oil of the cinnamon plant, has been demonstrated to have antimicrobial properties in multiple
in vitro studies. It is hypothesized that the cinnamaldehyde-eugenol-thymol blend and the lactoferrin tested in this study will
increase weight gain and decrease the incidence of diarrhea in veal calves. This study was conducted as a randomized controlled trial, with calves randomized to the lactoferrin, cinnamaldehyde, or control groups. Calves in the treatment groups
received 1g/day of either lactoferrin or the cinnamaldehyde blend. Primary outcome measures were body weight gain and
incidence of diarrhea. Secondary outcome measures included the incidence of respiratory disease and non-specific illness.
Health assessments were performed twice weekly for 21 days, and body weight was measured at 0, 21, 42, and 56 days. Differences in growth and the incidence of diarrhea between treatments will be compared using a generalized linear regression
model. The expected outcome is that calves supplemented with lactoferrin or the cinnamaldehyde blend will have increased
weight gain and decreased incidence of diarrhea as compared to the control group. The results of this study may indicate the
viability of using antibiotic alternatives as a treatment option for both veal and dairy producers.
Research Grant: Health Aging Nutrition, LLC, Columbus, IN
Student Support: Merial Veterinary Scholar program
216
Optimization of DNA Vaccine Targeting GnRH Receptor for Animal Contraception
Freelie L. Mitchell, Alexandre Samoylov, Anna Cochran, Bettina Schemera, James Wright, Tatiana Samoylova
Scott-Ritchey Research Center (Mitchell, A. Samoylov, Cochran, T Samoylova), College of Veterinary Medicine, Auburn
University, Auburn, AL 36849, USA; Laboratory Animal Health (Schemera), College of Veterinary Medicine, Auburn
University, Auburn, AL 36849, USA. Department of Pathobiology (Wright, T. Samoylova), College of Veterinary
Medicine, Auburn University, Auburn, AL 36849, USA.
1
Overpopulation of feral cats and stray dogs is a serious animal welfare as well as public health problem worldwide. Non-surgical, safe, cost-effective methods to reduce the number of animals born are urgently needed as a practical solution. The
focus of our research is on development of DNA-based contraceptive vaccines that target gonadotropin releasing hormone
receptor (GnRHR), a crucial player in animal reproduction. Previously, such vaccines were shown to stimulate immune
responses against GnRHR resulting in suppression of testosterone (indirect indicator of vaccine contraceptive value) in
immunized mice. While the desired functional effects were achieved, they appeared to be transient, most likely, because
amount of the expressed antigen was not high sufficiently and duration of the expression was not long enough to establish
immunological memory. The goal of the present study is to enhance and extend GnRHR expression for induction of strong
immunological responses leading to immunological memory and subsequent loss of fertility in immunized animals. Here,
we report the first step towards the goal - generation of two new DNA constructs with potentially improved properties. One
of the constructs contains an additional promoter (two promoters total) for increased antigen expression. The other construct contains CpG-free promoter for prevention of promoter methylation and expression silencing. The constructs were
designed, generated, and confirmed to have correct sequences. Successively, they will be tested for immune responses and
contraceptive efficacy in animals.
Research Grant: Found Animals Foundation, Michelson Grant in Reproductive Biology D1213-F13.
Student Support: Merial Summer Program, Scott-Ritchey Research Center, and Auburn University CVM.
Wil A Moorhead, Casey L Durfey, Gustavo D A Gastal*, Peter L Ryan, Scott T Willard, Jean M Feugang
Departments of Animal and Dairy Sciences (Moorhead, Durfey, Ryan, Willard, Feugang), Pathobiology & Population
Medicine (Ryan), Biochemistry, Molecular Biology, Entomology & Plant Pathology (Willard), Mississippi State
University, Mississippi; Department of Animal Sciences and Food nutrition, Southern Illinois University, IL
Two commonly assessed proteins in the swine industry, myoglobin (MYO) and fatty acid synthase (FAS), are considered
biochemical indicators of meat quality. MYO carries oxygen within tissues and gives meat its red color. FAS assembles the
various fatty acids that have differential effects on meat quality. The purpose of this study was to determine if these proteins
are expressed in muscle and fat tissues at different levels between genders. Gilts maintained in our experimental farm were
inseminated with extended boar semen and farrowing occurred at approximately 114 days post-insemination. At weaning,
twenty piglets (10 female and 10 male) were randomly selected and allowed to grow to market weight. Longissimus muscle
and subcutaneous back fat samples were collected and subsets were subjected to western immunoblotting (WIB) to confirm
specificity of protein targets using MYO and FAS antibodies. Remaining samples were prepared for immunofluorescence
(IF) detection. Standard protocols were used for both WIB and IF. Images were captured (EVOS microscope) and intensities were quantified (ImageJ). Data were analyzed (ANOVA/Wilcoxon) to compare protein fluorescence intensities within
tissues across gender. Results revealed high levels of MYO protein in male muscle and female fat. Females showed higher
muscular FAS content, while no significant difference was found in gender detection of FAS in fat tissues. These biochemical differences have implications on meat quality as differential FAS content could affect intramuscular fat accumulation or
marbling, a subjective indicator of palatability to consumers. The higher MYO content of male muscle will produce a darker
red meat, which is typically associated with greater tenderness.
Research Grant: USDA-ARS Grant#58-6402-3-018
217
Comparative immunogenicity of attenuated Salmonella vaccine strains dependent on parental origin and genotype
Leandra Mosca, Shilpa Sanapala, Roy Curtiss III
Department of Infectious Diseases and Pathology, College of Veterinary Medicine, University of Florida, Gainesville, FL
Since Salmonella is especially adept at invading and colonizing lymphoid tissue in the gut, it is considered the most
capable vaccine vector for oral immunization as a recombinant attenuated Salmonella typhimurium vaccine (RASV).
Prior work in our lab has suggested that the initial virulence, colonizing ability, and genotype of the RASV parental strain
is critical to the vaccine’s ability to elicit effective mucosal, cellular, and systemic immune responses and prevent infection by that bacterial pathogen. Salmonella vaccine strains of different parental origins and genotypes were compared for
their ability to elicit an immune reaction and for their effectiveness in protecting against challenge. Identical attenuation
means were used for the Salmonella typhimurium UK-1 strain, the most virulent strain known, and the 14028 strain that
is much less virulent. Identical attenuation means were also used for lysis and non-lysis RASVs x11829(pYA3342), and
for x11829(pYA4763) and x11803(pYA4763)(SifA+). We hypothesized that the more virulent Salmonella typhimurium
UK-1 strain would be more effective in inducing protection against challenge than the less virulent 14028 parental strain.
We also predicted that the lysis strain x11829(pYA3342) with arabinose-dependent murA expression would protect against
challenge more effectively than the non-lysis strain. Finally, we predicted that insertion of the normally-present sifA gene
back into the x11803(pYA4763) strain would decrease vaccine immunogenicity and therefore not protect as effectively
against challenge. These findings would provide basis on which to develop a more effective vaccine vector that minimizes
reactogenicity while increasing immunogenicity.
Research Grant: Uknown
Student Support: Merial, Florida Veterinary Scholars Program
Changes in periarticular bone secondary to osteoarthritis in chikungunya-infected and naive mice
Amber A. Moses, Brad A. Goupil, Christopher N. Mores, Heather Richbourg, and Margaret A. McNulty
Department of Comparative Biomedical Sciences (Moses, Richbourg, McNulty) and Department of Pathobiological
Sciences (Goupil, Mores), Louisiana State University School of Veterinary Medicine, Baton Rouge, Louisiana
Chikungunya virus (CHIKV) is a mosquito-borne alphavirus that is currently a threat to the U.S. due to a large outbreak in
the Caribbean and South America. The disease is associated with debilitating polyarthralgia, which has been shown to develop into chronic joint pain in 32-60% of those infected. Underlying joint disease has been associated with the development
of severe ongoing arthralgia following CHIKV infection. Previous work completed in this lab has characterized progressive
bone & joint lesions secondary to CHIKV infection in mouse models. The purpose of this study was to determine whether
CHIKV infection results in more severe disease in a model of post-traumatic osteoarthritis. We hypothesized that joints
surgically induced with OA and infected with CHIKV will have thicker subchondral bone and a reduction in epiphyseal
trabecular bone than control joints. Female C57/B16 mice underwent either surgical destabilization of the medial meniscus
(DMM) (n=31) or sham surgery (n=20) of the right hindlimb at 11 weeks old. One week after surgery, mice received either
an intra-articular inoculation of CHIKV (n=31) or a sham injection (SI) (n=20). Mice were sacrificed at 8 weeks (n=24) or
4 weeks (n=27) post-surgery. Hindlimbs were collected and routinely prepared for scanning with micro-computed tomography. Established parameters of bone volume and trabecular morphology were evaluated for the regions of interest: trabecular and subchondral bone in the proximal tibia. A significant difference in BV/TV in the proximal epiphyseal trabecular bone
of the tibia was identified at 8 weeks post-op (p=0.034); DMM+CHIKV mice was significantly higher than DMM+SI mice
(p=0.028). No significant differences were noted 4 weeks post-op.
Research Grant: NIH Centers of Biomedical Research Excellence Phase III Award 1P30GM110760-01 (CNM), NIH
5T32-oD011124-09 (BAG)
Student Support: Kenneth F. Burnes Trust
218
Moyer L, Tucker A, Gunter J, Bowser J, Cooley JA, and Swiderski CE
Mississippi State University CVM, Starkville, MS
Intradermal skin testing (IDT) is commonly applied in the diagnosis and management of allergic diseases in humans and
dogs. Allergen panels, developed and refined for human allergy, have been successfully adapted for the dog, a species that
cohabitates with man. Due to inconsistencies with testing outcomes, IDT has been widely criticized and considered of limited utility in horses. However, the environmental allergens of horses are radically different from humans and dogs.Insect bite
hypersensitivity, an IgE-mediated skin disease of horses, substantiates horses develop IgE sensitizing antibodies that arm
skin mast cells. Accordingly, we hypothesize that limited utility of IDT in horses is influenced by a bias towards antigens
of significance in human allergy, and an absence of replicates in IDT protocols from which to make valid assessments. To
test this hypothesis, we performed intradermal skin on 3 with pasture-associated asthma, and two non-diseased controls,
employing antigens of relevance to the environmental exposure of the horse. We performed replicate injections for each
antigen. Wheal size (mm)and induration (0-4) were evaluated at 20 minutes and 6 hours (reactions considered primarily
IgE- mediated) and also at 24 and 48 hours. Reactions were considered positive for an antigen when wheal and induration
were identified at all replicate sites. Consistent reactions to grass and mold antigens were identified within individual horses. IgE-mediated reactions were documented by response to replicate injections of Johnson Grass, Bipolaris spp, Meadow
Fescue, and Grass Smut at 20 minutes and 6 hours. We conclude that modifications of testing protocols for horses are likely
to improve the diagnostic utility of IDT.
Student Support: MSU College of Veterinary Medicine
Characterizing interactions between location, land use, and domestic dog health in Panamanian communities
S. M. Muller, J. L. Dyer, C. Mertzlufft, M. Perea, C. A. Rigg, I. Rodriguez, A. M. Santamaria, J. Tello, S. N. Tanner,
J. Velasquez Runk, J. E. Calzada, A. Saldaña, D. Alvarez, and N. L. Gottdenker
Department of Pathology, College of Veterinary Medicine (Muller, Gottdenker), Departments of Anthropology (Dyer,
Tanner, Velasquez Runk), and Geography (Mertzlufft), University of Georgia, Athens, GA. Instituto Conmemorativo
Gorgas de Estudios de la Salud (Perea, Rigg, Rodriguez, Santamaria, Tello, Calzada, Saldaña, Alvarez)
Human, animal, and ecological health interact in many ways. Research suggests that marginalized communities near areas
of recent deforestation and forest edges are at greater risk of exposure to zoonotic diseases. This study evaluates relationships between domestic dog health parameters and forest cover surrounding their home communities. We hypothesize that
dogs in geographically, socioeconomically marginalized communities near forests are in poorer overall condition and that
hematologic abnormalities are complexly related to deforestation due to variation in pathogen exposure. 277 dogs were
sampled from a total of six communities to the east (N=98) and the west (N=179) of the Panama Canal surrounded by one
of the following habitat matrices: primarily forested, fragmented forest, and deforested/pasture. We assessed the body condition score (BCS) of each dog and performed complete blood counts. 59.3% of dogs were underweight, with a significant association between habitat and BCS (N=273, Kruskal-Wallis x2=16.5, df=2, p<0.001). Forested habitats had the lowest mean
BCS (3.13, sd 1.68). We observed hematological abnormalities in 95% of dogs (N=222). The highest proportions of anemia,
leukocytosis, and thrombocytopenia were found in dogs living in fragmented forest, forested, and pasture habitats, respectively (67.0%, N=97; 65.8%, N=76; 82.1%, N=56). Dogs in communities east of the canal tended to have higher levels of
anemia and thrombocytopenia; leukocytosis was higher in western communities. Results suggest most dogs are exposed a
wide range of pathogens that differ based on habitat and geographic location. Ongoing studies assess household-level dog
health, integrating socioeconomic and zoonotic disease exposure data.
Research Grant: UGA-CDC Collaborative Research Seed Grant
219
Attempt to confirm absence of Avian Bornavirus in a previously shedding flock of cockatiels (N. hollandicus)
Ian Tizard, Olivia Murray, Kristen Streeter
The Schubot Exotic Bird Health Center (Tizard) Veterinary Pathobiology Department, Texas A&M University (Tizard),
College of Veterinary Medicine, Texas A&M, College Station, TX
Avian Bornavirus (ABV), the proven etiologic agent of Proventricular Dilatation Disease in parrots, is a major cause of concern in the avian health community. Due to intermittent shedding in the urates sampled via cloacal swab, as well as a lack of
alternative diagnostic methods, it has been unable to be determined whether or not affected birds are able to rid themselves
of the virus. For the last five years, the Schubot Exotic Bird Health Center at Texas A&M University has been acquiring
individual shedding data on a flock of sixty-six ABV infected cockatiels. They found that the occurrence of shedding tapered off after their first year at Texas A&M and eventually ceased. This lack of ABV positive cloacal swabs suggests that
cockatiels may be able to overcome ABV infection. To study this possibility further, seven birds were humanely euthanized
and necropsied according to an extensive pathogenesis protocol. Their tissues were tested for presence of ABV via two-step,
conventional Reverse Transcriptase-Polymerase Chain Reaction (RT-PCR) and all were negative for presence of the virus.
Immunohistochemistry results are still pending. However, due to the sensitivity of conventional RT-PCR and the extensive
sampling of the organs, it seems likely that the birds’ tissues are, in fact, rid of the virus. These findings, and future research,
may give hope to aviaries and aid veterinarians in the counsel of their clients with ABV infected cockatiels.
Research Grant: Richard M. Schubot Endowment
How to smoke your bacon: An investigation on the effects of environmental inhalants in a porcine model
Allison Mustonen, P. Sivasankar, A. Durkes
College of Veterinary Medicine (Mustonen, Durkes) and the Department Comparative Pathobiology (Durkes), Purdue
University, West Lafayette, IN; Department of Speech, Language & Hearing Sciences (Sivasankar), College of Health
Sciences, Purdue University, West Lafayette, IN
Approximately 7.5 million Americans suffer from difficulty using their voice. Phonatory difficulties are often multifactorial
with Reinke’s edema being one of the most common underlying pathologies. Common causes of Reinke’s edema include
smoking, voice overuse, and reflux of stomach acids into the larynx. The purpose of this study was to develop an in vivo
model of Reinke’s edema by challenging two group of six pigs with cigarette smoke in a customized inhalation chamber.
The first group of pigs was exposed to three cigarettes per day for 20 treatments (1 time per day, 5 days per week, 4 weeks).
The second group of pigs was exposed to five cigarettes per treatment for 60 total treatments (3 times per day, 5 days per
week, 4 weeks). A week prior to treatment, each group of pigs was habituated to the chamber and human contact to reduce
stress and ease treatment administration. Behavior modification was accomplished using positive reinforcement. At the end
of the four-week study, pigs were humanely euthanized and vocal fold, tracheal, and nasal mucosal samples were obtained
for histologic evaluation, RT-qPCR, and TEM. The week of habituation for each group of pigs successfully increased the
efficiency and reliability of treatments. The pigs willingly entered in the chamber and tolerated enclosure for the allotted
time. RT-qPCR and TEM are currently in progress. The data obtained from this study offers a potential novel experimental
methodology to simulate the development of Reinke’s edema in healthy pigs where there is currently no reproducible animal model. The success of this methodology could impact future research on prevention, improving early diagnosis, and
therapeutic options for Reinke’s edema.
Research Grant: NIH R01 DC011759
220
Characterizing infection of Jamaican fruit bats with bat-derived influenza-like viruses, HL17N10 and HL18N11
Laurel Myers, Kaitlyn Miedema, Ashley Malmlov, and Tony Schountz
Colorado State University College of Veterinary Medicine and Biomedical Sciences, Fort Collins, CO (Myers, Miedema,
Malmlov, Schountz); University of Wisconsin - Madison School of Veterinary Medicine, Madison, WI (Myers)
The genomes of two novel influenza-like viruses, HL17N10 and HL18N11, were recently identified in little yellow-shouldered bats (Sturnira lilium) and flat-faced fruit bats (Artibeus planirostris), respectively. Rescued viruses were categorized
as ‘Influenza A-like’ due to lack of canonical sialic acid receptor-binding activity and aberrant neuraminidase function.
These viruses have not yet been examined in their natural hosts. To this end, we studied the response of Artibeus jamaicensis
to inoculation with HL17N10 and HL18N11. Artibeus jamaicensis is a member of the same genus as A. planirostris, and the
same family as S. lilium; therefore, we used A. jamaicensis as a model. Three male bats were inoculated intranasally with
HL17N10, three with HL18N11. Nasal, oral and rectal swabs were collected for viral RNA (vRNA) isolation (via qRT-PCR)
and blood for ELISA. Animals were euthanized 28 days post infection (dpi) and tissues were collected for viral isolation
and histopathology. Daily temperatures remained within normal limits and animals showed no signs of clinical disease.
HL18N11 was identified from rectal swabs 2, 4 and 7 dpi, and, interestingly, HL17N10 was not detected at all. Serology
and histopathology results are pending. These data indicate that A. jamaicensis may be susceptible to HL18N11, but not to
HL17N10. The presence of HL18N11 vRNA in rectal swabs implies fecal-oral transmission, similar to avian influenza virus
transmission between birds. Further investigation is necessary to confirm this transmission route. Finally, it is imperative to
characterize neuraminidase-like enzyme activity in Influenza A-like viruses to better understand their replication cycle and
their inability to infect multiple species.
Research Grant: Startup funds to TS
Jonathan Nagel, Keijiro Mizukami, Karthik Raj, Anna Eringis, Noa Safra, Urs Giger
Section of Medical Genetics, School of Veterinary Medicine, University of Pennsylvania, Philadelphia, Pennsylvania
(Nagel, Mizukami, Raj, Eringis, Giger), School of Veterinary Medicine, University of California - Davis, Davis,
California (Safra)
Dog Erythrocyte Antigen 1 (DEA 1) can cause serious acute hemolytic transfusion reactions in DEA 1- dogs previously
transfused with DEA 1+ blood. About half of all dogs are DEA 1- and half are DEA 1+ (weakly to strongly positive). Thus
far, the DEA 1 protein and genetic basis have not been well defined. This study attempted to characterize the DEA 1 protein
by immunoblot and the DEA 1 gene by genotyping DEA 1- and weakly to strongly DEA 1+ dogs. Ghost cells were prepared
from a DEA 1+ and DEA 1- dog and used in immunoblot. To preserve the structure of the protein, the protein was exposed
to different conditions before gel electrophoresis (+/- heating, +/- reducing). Three bands at approximately 100, 70, and 40
kDa were found in samples from both the DEA 1+ and DEA 1- dogs. These bands are likely unspecific binding, and thus,
further conditions and immunoprecipitation will be tested to try to preserve the structure of the DEA 1 protein. To further
characterize the DEA 1 gene, DEA 1 typed dogs of different breeds were genotyped to investigate a SNP that appears to be
associated with DEA 1 in a previous genome-wide association study. With rare exception, DEA 1- dogs were homozygous
for C at this SNP, while DEA 1+ dogs were either heterozygous or homozygous for T at this SNP. These results confirm the
autosomal dominant inheritance of DEA 1 and that this SNP is a good genetic marker for separating DEA 1+ from DEA
1- dogs, but it does not differentiate the degree of DEA 1 expression. This SNP may be useful for determining the location
for the DEA 1 gene. In conclusion, advancements were made in characterizing the DEA 1 system in dogs, and additional
studies are in progress to further define DEA 1 at the molecular level.
Research Grant: NIH OD010939
Student Support: Supported by NIH grant T35 OD-010919 and a grant from Merial
221
Effect of Hypoxic Preconditioned Wharton Jelly Mesenchymal Stem Cell Exosomal miRNA
on Hypoxic Injured Neurons
Chrystal Nguyen, Asma Chaudri, Dawn Meola, Andrew Hoffman
Regenerative Medicine Laboratory, Cummings School of Veterinary Medicine at Tufts University, North Grafton, MA
Background: Stroke or spinal cord infarction patients often suffer neuronal damage from ischemic reperfusion. A possible
treatment could be stem cell therapy due to their protective and regenerative properties. Mesenchymal stem cells (MSC) are
known to secrete exosomes, nanoscale vesicles (~100 nm diameter) which contain proteins, mRNA, and miRNA known to
mitigate hypoxic ischemic injury. The effects of MSC are enhanced by exposure to low O2 levels (hypoxic preconditioning
- HP) which increases the release of anti-inflammatory, anti-apoptotic, and neuronal growth factors. It is unknown whether these effects are conveyed via exosomes. We hypothesize that subjecting canine Wharton Jelly MSC (WJMSC) to HP
will increase production of neuroprotective (NP) miRNA in exosomes, leading to improved therapeutic potential of MSC.
Methods: WJMSC were placed in 1-5% O2 to test viability. WJMSC were then exposed to 1% O2 for various times (24,
48, 72 hrs vs normoxia controls) and cellular miRNA (n=277) was analyzed. Results: HP WJMSC (vs normoxia) for up to
72 hrs maintain high viability (95-100%). Following HP, the concentrations of NP miRNA changes markedly compared to
normoxia. Of the top 25 upregulated miRNA, at 24 hrs, 3 are NP(miR-17,-210,-124); whereas at 48 hrs, 1 is NP (miR-223)
and 3 neuroinjurious (miR-29b,-140,-145); and at 72 hrs, 2 are NP (miR-210,-124). Conclusion: These data suggest that HP
for 24 or 72 hrs leads to an upregulation of NP miRNA without neuroinjurious miRNA. In the future, we will test miRNA
content of exosomes derived from HP WJMSC to assess their content of miRNA and their anti-apoptotic effects on oxygen
glucose deprived neuronal cell lines as an intermediate step before translation into animal models.
Research Grant: Shipley
Student Support: In part supported by a grant from Merial Veterinary Scholars Program
Steven Nguyen, Edward Vasquez, Oren Ofer, Pedro Paulo V. P. Diniz
College of Veterinary Medicine, Western University of Health Sciences, Pomona, California
Approximately 15% to 30% of all cats presented at a veterinary clinic may have undiagnosed cardiac disease and/or arrhythmias, which can only be precisely detected by electrocardiography (ECG). In 2013, a Smartphone-based ECG device
(AliveCor ) was introduced into both human and veterinary medicine, and is capable of recording ECG traces and storing
them online for local and long-distance review. Our objective was to evaluate the quality and accuracy of ECG traces from
cats generated by the smartphone ECG (hereafter “S-ECG”) when compared to standard six-lead computer-based ECG
(BASi Vetronics, hereafter “PC-ECG”). Thirty-four healthy client-owned cats were enrolled. Currently, ECG traces from
eleven out of 34 cats have been reviewed. Preliminary results indicate that ECG traces obtained from the S-ECG had better
quality than the PC-ECG. Body weight, percentage of body fat, coat type, and age did not significantly affect QRS amplitudes detected by the S-ECG. When comparing the MEA between the smartphone and the PC, the results did not correlate,
with 33% to 66% of cats being considered within normal limits by computer-based ECG but abnormal by smartphone ECG.
In conclusion, based on our preliminary results, the S-ECG seems to generate traces of similar or better quality from cats
than the traditional PC-ECG. Directional statistics of the total number of cats enrolled will confirm if the polarity of the
waves detected by S-ECG are significantly different from the PC-ECG. If the accuracy of traces is confirmed, our results
will support the use of smartphone ECG in feline medicine, which potential reduction in mortality due to occult cardiac
disease that would have gone undetected otherwise in a third of unscreened cats.
Research Grant: College of Veterinary Medicine Innovation of DVM Curriculum Grant, Western University of Health
Sciences.
Student Support: Merial- NIH Grant
222
The growing feather as a model for novel evaluations of Marek’s disease genetic resistance
Kayla Niel, Cari Hearn, and Hans Cheng
College of Veterinary Medicine, Michigan State University, East Lansing, MI (Niel) and United States Department of
Agriculture, Agriculture Research Service, Avian Disease and Oncology Laboratory, East Lansing, MI (Hearn, Cheng)
A recent assay has been developed using growing feathers (GF) to monitor immune responses of chickens. In the assay, test
samples are injected into GF and, at various times, the cellular pulp is analyzed. This assay could prove advantageous for
comparing immune responses of chickens that are genetically resistant or susceptible to Marek’s disease (MD), a lymphoproliferative disease induced by MD virus (MDV), an oncogenic herpesvirus. The GF model could eliminate the need to
sacrifice animals to access materials, while providing the ability to monitor an individual’s response to MDV over time. The
present study investigated the potential of this model for use in genetically unique populations at the ARS Avian Disease
and Oncology Laboratory to study MD resistance mechanisms. GF were clipped above the level of the epidermal cap and
injected with peptidoglycan, an antigen for Gram-positive bacteria, in MD resistant (line 63) and susceptible (line 72) hens.
GF were then collected over time, pulp from each GF extracted, and recovered cells immunofluorescently labeled for analysis by flow cytometry to determine resultant immune response profiles. Line 63 injected GF pulp had greater percentages
of lymphocytes overall, and CD4+ and CD8+ cells, than line 72. Percentages of KUL01+ cells were similar among 63 and 72.
These results suggest that there are distinctions in immune response at the feather follicle between MD resistant and susceptible birds to a common antigen, and that there may be important differences in their response to MDV. The success of the
GF as a dermal test site in this study suggests that this model may be useful for future studies to determine the influence of
immune response(s) on MD resistance.
Research Grant: USDA, ARS
Thermal nociception and gastrointestinal function following administration of Simbadol in rats
Nicole Nietlisbach, Molly Allen, Rebecca A. Johnson
Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI
Buprenorphine, a partial m-opioid agonist administered at 0.05 mg/kg subcutaneously every 6-8 hours, is a common peri-operative analgesic used in both laboratory and companion rats. This frequency of injection may increase patient stress and
discomfort, which may compromise research and be unsatisfactory to pet owners. A novel, high-concentration buprenorphine formulation, Simbadol , is effective in cats for up to 24 hours of peri-operative analgesia without major adverse effects
when administered at a high dose of 0.24 mg/kg (compared to the standard feline dose of 0.02 mg/kg) subcutaneously. In
a blinded cross over study, four doses of Simbadol (0.075, 0.15, 0.30, and 0.60 mg/kg) and a control (5% dextrose solution) will be administered subcutaneously to 8 male Sprague-Dawley rats with a one-week washout period between treatments. Anti-nociception will be assessed by Hargreaves plantar thermal latency testing and gastrointestinal function will
be assessed by measuring food intake, fecal number, and fecal mass. Thermal latency measurements will be taken before
treatment and at 1, 4, 8, 12, 24, and 36 hours after treatment. Food intake and fecal output will be measured every 12 hours
for 36 hours before and 36 hours after treatment. Rats will be housed singly in wire-bottom cages to facilitate measuring
food intake and fecal output. We hypothesize that Simbadol will be effective (prolonged thermal latency) in rats for up to
24 hours, albeit with some degree of reduced gastrointestinal function. Provided any adverse effects are within acceptable
limits, we anticipate that the results of this study will implicate Simbadol as an alternative for improved peri-operative pain
management in research and companion rats.
Research Grant: Companion Animal Grant- University of Wisconsin
Student Support: University of Wisconsin- Madison
223
Discovery of common genetic risk loci for canine cruciate ligament rupture by validation GWAS
Jacob Nilles, Lauren Baker, Emily Binversie, Alex Piazza, Guilherme Rosa, Brian Kirkpatrick, Daniel Gianola,
Susannah J Sample, Peter Muir
Department of Surgical Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI
Canine non-contact cruciate rupture (CR) has breed and familial predispositions and is common, representing about 20% of
canine lameness. CR is a complex trait disease with genetic and environmental factors. We hypothesize the genetic contribution to CR in dogs is comprised of many risk loci with small additive effects, including common loci in different breeds
of dog. We conducted a validation genome-wide association study (GWAS) in Rottweiler dogs with and without CR (n=92)
to study CR genetics. Genotyping used the Illumina CanineHD Beadchip single nucleotide polymorphism (SNP) array. We
studied 74 cases, 18 controls, 41 males and 51 females. Preliminary data analysis suggests that two risk loci exist on CFA4
in the Rottweiler, which contain 19 genes. In an initial discovery GWAS in the Labrador Retriever, we identified 99 candidate loci in the genome associated with CR. This suggests that CR is a highly polygenic disease. The associated regions on
CFA4 in the Rottweiler are distinct from the Labrador Retriever loci, suggesting that disturbances to biological pathways
that contribution to CR pathogenesis may differ between breeds of dog. The polygenic nature of CR suggests that clinical
genetic testing will need to use genomic prediction by whole genome regression. Our data suggest that it may be difficult
to perform between-breed prediction. Our Rottweiler recruitment is ongoing and we expect to have recruited 200-300 dogs
by the end of the validation GWAS.
Research Grant: Melita Grunow Family Professorship in Companion Animal Health
Student Support: School of Veterinary Medicine, University of Wisconsin-Madison
A proposed method for functional quantitation of carpal joint laxity in canine mucopolysaccharidosis type I
Thomas F. Oelfke, Andrew Hess, N. Matthew Ellinwood
College of Veterinary Medicine (Oelfke, Ellinwood), Department of Animal Science (Hess, Ellinwood), College of Agriculture and Life Sciences, Iowa State University, Ames, IA
The mucopolysaccharidoses are rare inherited lysosomal storage diseases of both humans and canines, in which individuals
lack specific enzyme activity in the degradation pathway of glycosaminoglycans (GAGs). Human mucopolysaccharidosis
type I (MPS I) patients have progressive connective tissue and bone disease including joint restriction. While much of the
pathology in the canine model is similar, canines are unusual in that they have profound joint laxity, most notable as carpal
hyperextension. The aim of our study is the quantification and comparison of carpal joint extension in both normal and MPS
I affected dogs. Kinematic data capture will use infrared sensors, reflective markers, and tracking software. Eight infrared
cameras (Vicon ; Oxford Metrics, Oxford, UK) will be used to track the positions of four 15mm optical markers attached
to specific points on each of the animals’ forelimbs with adhesive Velcro . Markers will be placed which define the radius
and the the fifth metacarpal. Coordinates x, y, and z of the optical markers will capture the course of a walking gait on a
treadmill. The linear projections of the limb segments proximal and distal to the carpal joints will be mapped over time to
find their maximum angle of intersection. Data will be analyzed by repeated measure ANOVA. Twenty-four subjects (14
male and 10 female, most of which are sibling pairs (22)) of which 12 are MPS I affected and 12 are unaffected controls
will be evaluated. Ages range from 3 to 50 months of age. The development of a fully quantitative method to assess carpal
hyperextension in the MPS I dog model will provide a rigrorous pre-clinical outcome mearsure by which to evaluate further
improvements in therapy for this disease.
Research Grant: NIH R01NS085381 Awarded to Patricia I Dickson
224
Oren A. Ofer, Edward Vasquez, Steven Nguyen, Pedro Paulo V. P. Diniz
Electrocardiography (ECG) remains the gold-standard test for arrhythmogenic cardiac disease. However, the use of current
ECG devices in veterinary medicine has several limitations: cost, equipment size, lead wires, and data-sharing ability across
various platforms. Our objective was to evaluate the use of a smartphone-based ECG device (AliveCor , hereafter “S-ECG”)
in dogs. Thirty-five client-owned dogs of different breeds, ages and coat-types were enrolled. S-ECG recordings were analyzed and compared to standard 6-lead computer-based ECG (BASi Vetronics, hereafter “PC-ECG”) traces. The quality
of each was graded on a 1-5 standardized scale by each investigator. Currently, ECG traces from ten out of 35 dogs have
been reviewed. Preliminary results indicate no significant differences in ECG quality between S-ECG and PC-ECG for three
different body positions. When the mean electrical axis (MEA) was classified as normal or abnormal, 30-45% of dogs with
an MEA considered normal on the PC-ECG had their values shifted to abnormal when measured by S-ECG. Body weight,
percentage of body fat and age did not interfere with QRS amplitudes detected by S-ECG. Amplitudes of QRS complexes
were significantly lower in dogs with wire or long hair than in dogs with short hair when recorded with the S-ECG. This
suggests that coat type may interfere with the detection of cardiac electricity. Directional statistics will confirm if wave polarity detected by S-ECG is significantly different from PC-ECG. Once traces of all enrolled dogs are analyzed, our results
may support the use of smartphone ECG in canine health, which could potentially benefit many dogs in shelters, rural areas,
ambulatory clinics, and military or law-enforcement.
Research Grant: College of Veterinary Medicine Innovation of DVM Curriculum Grant, Western University of Health
Sciences
Development of large animal models for a compound heterozygous human disease using CRISPR/Cas9
James L. Oldeschulte, Carlos A. Pinzon, Diarra Williams, Shannon Huggins, Larry J. Suva, and Charles R. Long
Department of Veterinary Physiology and Pharmacology, College of Veterinary Medicine, Texas A&M University,
College Station, TX
Hypophosphatasia (HPP) is a rare inborn error of metabolism caused by mutations in the tissue-nonspecific alkaline phosphatase (TNSALP) gene that present with bone and tooth abnormalities. Many of the viable mutations are loss-of-function
missense mutations in one of the 12 exons of TNSALP. Current murine models harboring TNSALP mutations do not accurately represent the spectrum of TNSALP function, as mouse and human serum levels are not comparable. Since sheep
and humans have similar serum TNSALP levels, and bone remodeling is analogous, we hypothesized that targeting specific
gene regions in isolated sheep fibroblasts using CRISPR/Cas9 would model different allelic variants associated with HPP.
To accomplish this, 14 short guide RNAs targeting four unique regions in three different exons of the sheep TNSALP gene
were designed using bioinformatic web tools. Primers to capture the regions 2 kb up and down-stream of these targets were
designed using the NCBI primer-blast tool and the ThermoFisher Multiple Primer Analyzer. All guide RNAs were cloned
into the Px459v2.0 plasmid. Sheep fibroblasts were transfected with 11 of the guides using Lipofectamine 3000. A T7
endonuclease assay will be performed to determine the effectiveness of the guides. Once optimized, the different sgRNAs
and DNA templates containing orthologous HPP mutations will be used to modify sheep embryos by intracytoplasmatic
microinjection. This will create four unique allelic variants of HPP representing the most common, yet viable, mutations at
the TNSALP gene. These variants will be used to model the human compound heterozygous HPP phenotype in sheep.
Research Grant: LINK Equine Research Endowment
225
Erika D. Olney, Caroline Andrews, Scott K. Durum
Cancer and Inflammation Program, National Cancer Institute, Frederick, MD
Inflammatory bowel disease (IBD) is a debilitating group of conditions thought to result from loss of epithelial barrier integrity and subsequent inappropriate exposure of intestinal immune cells to luminal antigens. Interleukin-27 (IL-27) is a promising potential therapy for IBD, having demonstrated efficacy in three murine colitis models. Expression of both subunits of
the IL-27 receptor (IL-27R) has been demonstrated in epithelial cells from human colon biopsies and colorectal cancer cell
lines; however, its expression has not been characterized in the intestinal epithelium of mice, which serve as valuable models for IBD research. The objectives of this study were to investigate the expression of the IL-27R in murine small and large
intestinal epithelial cells and to characterize how inflammation may alter this expression. Small and large intestinal crypts
from C57BL/6 mice were cultured in vitro to produce intestinal organoids. Organoids were stimulated with IL-27, interleukin-1b, tumor necrosis factor-a, interferon-g, or endotoxin for 2, 24, and 48 hours, followed by RNA extraction and real
time quantitative PCR analysis for each subunit of the IL-27R (IL-27Ra and gp130). Complementary to previous findings
in our laboratory that IL-27 did not directly activate murine small intestinal epithelial cells within 60 hours, preliminary data
suggests that unstimulated small intestinal organoids or those incubated under the conditions above lack IL-27R expression.
Data for large intestinal organoid expression of IL-27R is pending. These data provide insight into interactions between the
intestinal epithelium and immune system and inform future investigations into the role of IL-27 in intestinal inflammation.
Research Grant: Intramural Research Program of the National Institutes of Health, National Cancer Institute
Student Support: National Cancer Institute
Seasonal variation of 25-hydroxy-vitamin D in two captive Eastern black rhinoceros (Diceros bicornis michaeli)
Wes Oltman, June Olds, Drew Makowski, Lou L. Keeley, and Ju Ji
Department of Veterinary Clinical Sciences, Iowa State University College of Veterinary Medicine, Ames, Iowa (Oltman,
Olds); Heartland Assays, Iowa State University Research Park, Ames, Iowa (Makowski); Blank Park Zoo, Des Moines,
Iowa (Olds, Keeley); Department of Statistics, Iowa State University, Ames, Iowa (Ji)
The black rhinoceros (Diceros bicornis) is a critically endangered species, with approximately 65 individual animals housed
in captivity within AZA-accredited zoos in the United States, and less than 5500 individual animals of all subspecies surviving in the wild. Previously published reference values in Diceros spp. for 25OHD3 [55.7 (+/- 34.2) ng/ml] were based
upon a small number of free-ranging black rhinoceros [n=28]. Recent human research has highlighted the importance of
sub-clinical vitamin D deficiency with links to increased risks for developing a variety of diseases such as cancer, autoimmune conditions, cardiovascular disease and infectious diseases such as tuberculosis. Subclinical vitamin D deficiency
is also likely to reduce breeding success in animals through diminished fertility and a reduced ability to maintain calcium
levels sufficient for normal fetal growth. Serum samples were collected opportunistically from two captive Eastern black
rhinoceros (Diceros bicornis michaeli) housed with seasonal access outdoors in a North American zoo to test for 25-hydroxy-vitamin D (25OHD) levels over a three-year period. A commercially prepared pelleted diet which included dietary
vitamin D3 was also fed to both rhinos. This study correlates environmental UVB, dietary supplementation, and seasonal
serum 25OHD levels to compare to known 25OHD3 levels in free-ranging African black rhinoceros. Results from these two
individual animals suggest that Diceros bicornis are dependent upon sunlight or UVB for measurable circulating 25OHD
and that dietary supplementation has little to no effect in captive Diceros housed in northern latitudes. Additional research
is needed to understand the function of 25OHD in Diceros spp.
Research Grant: Blank Park Zoo
Student Support: Merial and the Iowa State University College of Veterinary Medicine
226
Marble Burying for Assessing Postoperative Pain in Rats Treated with Meloxicam or Sustained-Release Meloxicam
Robert K. Onaga and Kristopher G. Galang, Jessica D. Ayers, and Lon V. Kendall
Laboratory Animal Resources, Department of Microbiology, Immunology, and Pathology, College of Veterinary
Medicine and Biomedical Sciences, Colorado State University, Fort Collins, Colorado
Rats are a common model for research studies involving the use of pain-inducing events that must be ameliorated with analgesics. Rats bury novel objects as a function of their natural behavior to new objects and is a common method to evaluate
anxiety in rats. This study investigates marble burying as a unique means to assess pain in laboratory rats. We hypothesize
that rats in pain will bury less marbles. Male Sprague-Dawley rats were castrated or received anesthesia only and divided
into 3 respective treatment groups of 6 rats each: meloxicam (2 mg/kg SC once daily for 3 days), sustained-release meloxicam (SRM; 4 mg/kg SC once), or saline. Assessments were done at baseline and 1, 6, 12, 24, and 48 h post-surgery. Rats
were observed for 5 minutes to evaluate behavioral indicators of pain including orbital tightening, wound licking, grooming,
and rearing. Then, rats were placed in individual marble-containing cages and given 30 m to bury marbles. Castrated rats
given SRM had the highest rearing activity post-surgery, while castrated rats given meloxicam had the highest grooming
and the least wound licking activities post-surgery. Castrated rats in the saline treated group buried fewer marbles and had
the lowest magnitude increase in marble burying from 1 h to 48 h post-surgery as compared to castrated rats that received
analgesics and the anesthesia-only groups. This suggests the marble burying test may be used as an adjunct to other pain
assessment indicators to identify pain in rats.
Student Support: ASLAP Foundation
Licorice root: a phytoestrogen supplement with neuroprotective effects on cognition
Caitlin E. Ondera, Payel Kundu, William G. Helferich, Donna L. Korol, Ikhlas A. Khan, and Susan L. Schantz
Dept of Comp Biosci (Ondera, Schantz), Neurosci Prog (Kundu, Schantz), Beckman Inst (Schantz), and Dept of Food
Sci & Human Nut (Helferich), University of Illinois Urbana-Champaign, Urbana, IL; Dept of Biol (Korol), Syracuse
University, Syracuse, NY; Natl Ctr for Natural Prod Res (Khan), University of Mississippi, University, MS
Licorice root (Glycyrrhiza glabra) is a dietary supplement that is widely used for the treatment of menopausal symptoms
in women. It contains the phytoestrogens liquiritigenin and isoliquiritigenin that exhibit estrogenic activity. Phytoestrogens
are a promising alternative to estrogen replacement therapy, which increases the risk of cancer, stroke, and cardiovascular
disease, but their efficacy and safety are not well understood. Estradiol administration to ovariectomized rats improves performance on hippocampal-dependent tasks but causes deficits on prefrontal and striatum-dependent tasks. Conversely, our
previous research showed isoliquiritigenin improves performance on a hippocampal task and has no effect on a prefrontal
task. This study focused on the effects of licorice root on the striatum to determine if it causes deficits similar to estradiol.
Licorice root powder, licorice root extract, and isoliquiritigenin were administered orally to ovariectomized Long Evans
rats. Estradiol positive and negative control groups were also included. After 3 weeks of exposure, rats were subjected to a
striatum-dependent object recognition task. Rats were placed in a testing chamber and allowed to habituate to two stimulus
objects. The familiar objects were then replaced with two novel objects. Object exploration time was recorded, and a pattern
separation index was calculated between habituation and test trials. Rats that recognize the second pair of objects as novel
have a greater pattern separation index, indicating better performance. This work may lead to the identification of a dietary
supplement for treatment of menopausal symptoms that confers neuroprotective effects without the negative effects of other
estrogens.
Research Grant: NIH, 5 P50 AT006268-06
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Sean Pador, Jeeva Subbiah, Brittany Voss, and Mohammad Mir
College of Veterinary Medicine, Pomona, CA
Hantaviruses are rodent borne, enveloped, negative sense RNA viruses of the Bunyaviridae family. The hantavirus genome
is comprised of three RNA segments: Small (S), Medium (M), and Large (L). S encodes the nucleocapsid protein (N), M
encodes the glycoprotein precursor, and L encodes the RNA dependent RNA polymerase. While rodent infections are asymptomatic, human infections may result in hantavirus cardiopulmonary syndrome and/or hemorrhagic fever with renal
syndrome. Currently, no treatments are available for this viral infection. N preferentially facilitates the translation of viral
mRNA in host cells. N binds to both the 40S ribosomal subunit and viral mRNA 5’ UTR. N associated ribosomes are loaded
on viral transcripts to boost their translation in infected cells. Our lab has previously identified a chemical compound (K31)
that interrupts the interaction between N and viral mRNA 5’ UTR. A single dose of this inhibitor inhibits viral replication
by 95 percent within 24 hours post infection. To synthesize more potent derivatives of K31, a co-crystal structure of the
N-K31 complex needs to be resolved. The wild type N undergoes aggregation upon purification and thus poses problems
upon crystallization. However, it has been reported that N mutants having deletions at the N and C terminal regions do not
aggregate. I cloned, expressed and purified an N protein mutant having deletions at N and C terminal regions. The purified
protein will be tested for K31 binding and used to solve the K31-N co-crystal structure. This structure will play a vital role
in the synthesis of new derivatives having high target binding affinity and improved efficacy as antiviral therapeutics.
Research Grant: Merial Veterinary Scholars Program, Western University of Health Sciences Intramural Funding
Student Support: Merial Veterinary Scholars Program, Western University of Health Sciences Intramural Funding
Efficacy of a new diagnostic procedure, the Mini-FLOTAC, in the diagnosis of canine Giardia infection
Lauren Page, Meriam Saleh, Katelynn Monti, Brian Herrin, and Anne Zajac
Virginia-Maryland College of Veterinary Medicine, Virginia Tech, Blacksburg, VA (Page, Saleh, Monti, and Zajac),
Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK (Herrin)
The Mini-FLOTAC is a new diagnostic procedure used in the microscopic examination of feces for detection of parasitic
eggs and cysts. While its efficacy for diagnosing Giardia duodenalis infections in humans has been studied, little is known
about its efficacy in recovering canine Giardia cysts. Previous studies have indicated that zinc sulfate centrifugal fecal flotation is the most sensitive microscopic procedure for detecting G. duodenalis cysts in veterinary clinical practice. However,
many practitioners continue to use passive fecal flotation procedures. Therefore, centrifugal fecal flotation was used as
a reference to compare mini-FLOTAC and passive fecal flotation procedures, in order to determine if the mini-FLOTAC
could provide greater sensitivity for G. duodenalis detection than passive flotation without requiring a centrifugation step.
An overall G. duodenalis prevalence of 8.77% was found in the 114 canine samples that were evaluated, based upon the
reference procedure. The sensitivity for passive fecal flotation compared to the reference test was 60%, and the sensitivity
for the mini-FLOTAC compared to the reference test was 70%. Both had a specificity of 100%. Sensitivities were compared
using McNemar’s test for significance of differences, and there was no statistically significant difference in sensitivities between the two tests. The results of this study do not support the idea that the mini-FLOTAC is significantly more sensitive
than a passive fecal flotation in G. duodenalis cyst recovery.
Research Grant: Virginia-Maryland College of Veterinary Medicine
Student Support: Virginia-Maryland College of Veterinary Medicine
228
In vivo and in vitro evaluation of mesenchymal stem cell therapy for primary osteoarthritis
Stephen C. Pannone, Valerie Johnson, Aimee C. Colbath, Kelly S. Santangelo and Steven W. Dow
Department of Microbiology, Immunology and Pathology (Pannone, Santangelo), Colorado State University, Fort Collins,
CO, Department of Clinical Sciences (Johnson, Colbath, Dow), College of Veterinary Medicine, Colorado State
University, Fort Collins, CO
Primary osteoarthritis (OA) is a debilitating disease associated with pain and decreased range of motion. This condition results from irreversible degradation of cartilage, intermittent soft tissue inflammation, and remodeling of subchondral bone.
Current therapy is limited to pain management, lifestyle changes, and joint replacement. Administration of mesenchymal
stem cells (MSCs) may be a promising cellular therapy for OA due to the anti-inflammatory and regenerative properties
of these cells. Our laboratory has demonstrated that pre-activation of MSC (aMSC) with an inflammatory stimulus may
increase the anti-inflammatory and immunomodulatory capacity of these cells. To investigate this novel treatment we will
evaluate intravenous administration of MSCs in a spontaneous model of OA and evaluate the ability of these cells to alleviate systemic signs of OA. Our hypothesis is that intravenous MSCs, particularly aMSCs, will ameliorate clinical signs and
decrease levels of inflammatory mediators in the joints of guinea pigs with chronic OA. To evaluate efficacy of this therapy,
we will administer intravenous resting or aMSCs to animals and monitor tracking to joints via IVIS imaging. The effect of
both resting and aMSCs on chondrocytes will also be examined in vitro to elucidate mechanisms involved in this therapy.
Cultured chondrocytes, stimulated with inflammatory mediators and pre-incubated with MSC or aMSC conditioned media,
will be examined to evaluate changes in cytokine production and inflammatory gene expression. These studies are critical to
improve the current treatment of this debilitating disease, as well as to elucidate the mechanism by which these cells exert
an effect on the local environment of the joint.
Research Grant: Shipley Foundation
Student Support: NIH T35 Training Grant NIH T35OD015130
Assessment of cellular response to synovial joint fluid in osteoarthritic and healthy dogs
Delaney F. Patterson and Aaron M. Stoker
Comparative Orthopaedic Laboratory, College of Veterinary Medicine, University of Missouri, Columbia, Missouri
Osteoarthritis (OA) is a debilitating disease that impairs movement in both dogs and humans. Synovial fluid lubricates the
joint and is the mechanism for avascular cartilage tissue to obtain nutrition and remove waste, and transmits signaling peptides of inflammation and degradation between the tissues of the joint associated with OA development and progression.
Therefore, for this study the cellular response of chondrocytes, synoviocytes, and meniscal fibrochondrocytes to synovial
fluid obtained from dogs with and without OA will be analyzed. Our hypothesis is that the gene expression of monolayer
cell cultures exposed to OA synovial fluid will significantly increase for degradative enzymes and inflammatory biomarkers,
and significantly decrease for extracellular matrix proteins compared to cells exposed to healthy synovial fluid. With ACUC
approval, synovial fluid was collected from 5 healthy dogs euthanized for reasons unrelated to this study and 6 dogs undergoing a surgical procedure for OA of the stifle at the MU Veterinary Health Center. Primary cell cultures of chondrocytes,
synoviocytes, and meniscal fibrochondrocytes were created from healthy dogs euthanized for reasons unrelated to this study.
Cells were cultured on 24 well plates in 0.5 ml of media containing 10% normal or OA synovial fluid. Control cells were
not exposed to synovial fluid. Cells were cultured for 24 or 72 hours and then total RNA was extracted and relative gene expression was determined by real time RT-PCR. Genes associated with inflammation, tissue extracellular matrix production,
and degradative enzyme production and regulation will be assessed and presented for each cell type.
Research Grant: Comparative Orthopaedic Laboratory, University of Missouri
Student Support: Endowment established by IDEXX-BioResearch
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Effects of intravenous fluid therapy on serum osmolarity in hospitalized dogs
Cristian Perez, Thomas Schermerhorn
Department Clinical Sciences (Perez, Schermerhorn), College Veterinary Medicine, Kansas State University, Manhattan,
KS
Serum hypertonicity is defined by an elevated concentration of solutes that regulate water movement across cell membranes.
Sodium (Na) and glucose (GLU) are the major endogenous solutes that affect serum tonicity (OsmE). A hypertonic state
has been associated with a variety of conditions (geriatric age, obesity, diabetes) and may affect disease progression and
prognosis. A better understanding of hypertonicity in dogs could allow clinicians to better manage fluid balance and impact
clinical decisions regarding fluid therapy. The objective of the study was to examine how intravenous fluid therapy affects
serum tonicity in hospitalized dogs. A prospective study design compared serum osmolality, pertinent solute concentrations
and other indicators of tonicity in clinically-ill dogs (n= 14) receiving intravenous isotonic crystalloid fluid for 48 hrs. Blood
samples for complete blood count, serum chemistry and serum total osmolarity (OsmT) were collected before (pretreatment) and 24-hr and 48-hr after continuous intravenous administration of isotonic fluid (0.9% NaCl or Lactated Ringer’s
Solution). The results (means are shown) reveal that OsmT decreases over 48-hrs (DOsmT = -8 mOsM). Changes in OsmT
were mostly due to changes in serum Na and blood urea nitrogen (BUN). Serum Na increased (DNa = 3 mEq/L) during the
48-hr period while BUN decreased (DBUN = -16mg/dl). Any increase in Na also increases OsmE. A decrease in BUN does
not affect OsmE but decreases OsmT. In conclusion, fluid therapy is associated with Na gain but a decrease in OsmT. The
observed OsmT decrease exceeds the BUN decrease and may reflect loss of unmeasured osmoles. Future work will examine
the clinical impact of Na and OsmT changes in hospitalized dogs.
Research Grant: Funded by Department of Clinical Sciences Research Grant
Student Support: CEVA Biomune.
The impact of mastitis on maternal behavior in dairy cows: a pilot study
Nadege Perier, Margit Bak Jensen, Deanna Marie Trearchis, Emma Bratton, and Kathryn Proudfoot
Vetagro Sup, Lyon, France (Perier), The University of Aarhus, Denmark (Jensen), The Ohio State University CVM,
Columbus, OH (Trearchis, Bratton, Proudfoot)
Dairy cows are at high risk of becoming ill after calving. Sick cows behave differently than healthy cows, but no research
has studied the effect of illness on maternal behavior. The objective of this study was to conduct a pilot study assess the
effect of mastitis on maternal behavior after calving. Ten multiparous Danish Holstein dairy cows were included in this
study. After calving, all cows were housed in individual pens with their calves for 4 d, and were screened for mastitis twice
daily. All calves were provided 4 L of colostrum by bottle within 6 h of birth. Five cows had been diagnosed with mastitis
within 4 d of calving; these cows were matched for parity with five healthy cows. Dam grooming behavior, and the amount
of time the calf spent drinking colostrum (from the dam or bottle) and searching for the udder, was measured continuously
by video recording for 24 h after calving. Due to the low sample size, descriptive data will be presented. Healthy cows
spent an average of 177.6 6 44.4 min/24h (mean 6 SD) grooming their calves, whereas mastitic cows spent 130.3 6 22.2
min/24h grooming. Calves from healthy dams spent about 28.4 6 23.7 min/d drinking colostrum, and about 4.6 6 1.9 min/d
searching for the udder. Calves from mastitic cows spent 31.04 6 18.2 min/d suckling, and 12.3 6 7.1 min/d searching for
the udder. The preliminary results from the study suggest that there may be some differences in behavior between mastitic
and healthy cows, but a larger sample size is needed to confirm these differences. Changes in maternal behavior after calving
may be a useful early indicator of mastitis, and may impact the way that producers manage calves from sick dams if they
choose to keep them together after calving.
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Neutralization of chlorhexidine-containing products in a clinical hand hygiene efficacy study
Laura R. Perry, Matthew Saab, Nora Biermann, J T. McClure and Aimie Doyle
Department of Health Management, Atlantic Veterinary College, University of Prince Edward Island, Charlottetown, PE
Surgical hand preparation products containing chlorhexidine-gluconate (CHG) have been proposed to need neutralization
in efficacy studies because of residual activity post scrub/rub. A neutralizing solution (NS) has been suggested for either
collection or dilution solutions. The addition of NS directly to surgeon’s hands in a clinical setting is undesirable, potentially
increasing patient risk of post-surgical complications. The objective of this study was to determine if dilution solutions with
NS effect bacterial counts after hand preparation using products containing CHG in an equine clinical setting. Hand preparations were randomly assigned each week: 4% CHG brush (CHG scrub), 4% CHG rub, 61% ethanol/1% CHG rub (EtCHG),
and 30% 1-propanol/45% 2-propanol rub. Surgeons were sampled pre and post scrub/rub and at the end of surgery using the
glove juice method. Aliquots were immediately diluted in PBS tween (PBST) with and without NS and plated in duplicate
within 3 hours to 3M Petrifilm Aerobic Count (AC) Plates. Bacterial counts were performed using a petrifilm reader. Data
were analyzed using a linear mixed model where the log-average difference was the outcome of interest and surgeon was
included as a random effect. Significance was set at p#0.05. Five surgeons participated and samples from 64 surgeries were
included for analysis. Bacterial counts were significantly higher in the NS samples post scrub from hands prepared with
CHG scrub (p=0.001) and in NS samples at the end of surgery for hands prepared with CHG scrub (p=0.045) and EtCHG
(p<0.001). Further work investigating the efficacy of each hand preparation product and the impact of neutralizer on CHG
products post collection in a clinical setting is ongoing.
Research Grant: Boehringer Ingelheim Equine Research Award and Atlantic Veterinary College internal grant
Student Support: Boehringer Ingelheim Equine Research Award and Atlantic Veterinary College internal grant
Role of Mouse Sca1+ Lung Mesenchymal Stem Cells in Bacterial Pneumonia
Elsie L. Phillips, Tirumalai Rangasamy, Shanshan Cai, Sagar Paudel, Laxman Ghimire, and Samithamby Jeyaseelan
Lung Biology Laboratory, Department of Pathobiological Sciences, School of Veterinary Medicine, Louisiana State
University, Baton Rouge, Louisiana
Pneumonia affects approximately 450 million people in the world and results in about 4 million deaths a year. In the United
States, nearly 3 million people are affected yearly with 5% of that population succumbing to the disease. One of the dangers associated with bacterial pneumonia is the possibility for the progression to sepsis, a condition marked by a dramatic
upregulation of the body’s immune response. Klebsiella species are a remarkably common isolate in intensive care units,
with Klebsiella pneumoniae frequently responsible for pneumonia based mortality in both immunocompetent and immunocompromised individuals. The increasing prevalence of antimicrobial drug resistance, such as carbapenem-resistant K.
pneumoniae, and a lack of effective vaccines has led to a dire need for novel treatments of bacterial based pneumonia. Past
studies have demonstrated that bone marrow derived stem cell therapy has the potential to both modulate and refine the
immune response generated during bacterial pneumonia. Despite this, little has been done to examine to potential benefits of
lung derived mesenchymal stem cells (LMSCs). This in vitro study has found that the presence of LMSCs causes a marked
decrease in inflammatory cytokines produced by both macrophages and neutrophils, following exposure to K. pneumoniae.
Additionally, it was determined that LMSC have the potential to induce more effective bacterial killing, either alone or in
combination with neutrophils. These finding have solidified the potential for lung mesenchymal stem cell based therapies
for bacterial pneumonia, establishing a baseline for future in vivo studies.
Research Grant: NIH pilot grant COBRE p30 GM110760-02 NIH grants 2RO1HL-091958 and 1RO1AI113720-01A1
Student Support: NIH T35 4T35OD011151-13
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Investigation of novel chemotherapeutic therapies for feline oral squamous cell carcinoma
Hunter Piegols, Maciej Parys, Marilia Takada, Taylor Aiello, and Vilma Yuzbasiyan-Gurkan
College of Veterinary Medicine, Michigan State University, East Lansing, MI
Feline oral squamous cell carcinomas (SCC) are highly aggressive neoplasms with short survival times despite multimodal
treatment. We hypothesize that more effective therapeutic strategies can be developed which target the bulk of the tumor as
well as putative stem cell fractions and increase the radiosensitivity of neoplastic cells. The current study seeks to identify
novel chemotherapeutics that fulfill these criteria. We previously performed a high throughput drug screen with over 2,000
drugs. Two of the drugs identified in the drug screen and two drugs with similar molecular mechanisms to drugs found to
be efficacious in the screening were selected for this study. We hypothesized that these drugs will inhibit the growth of three
SCC cell lines: SCCF1, 2, and 3 (kindly provided by Dr. T. Rosol of OSU) at levels achievable in plasma. Drug inhibition
profiles identifying the IC50 were generated for each drug and cell line using MTS assay. In addition, the effects of the drugs
of interest on cell cycle progression were analyzed via propidium iodide DNA labelling and flow cytometry to provide insight into the modes of action. Our findings show effectiveness of at least one of the drugs at low nanomolar concentrations.
With two of the drugs, evidence for arrest in G1 and G2 was found in the cell cycle studies. Investigations into the molecular
targets of each drug and the ability of the drugs to increase the radiosensitivity of SCC cell lines and target putative stem
cell fractions are under progress. Through these studies, our ultimate goal is to develop novel therapeutic strategies for the
treatment of SCC to provide better care for affected cats which can also inform translational studies in human squamous
carcinoma.
Research Grant: Endowed Research Funds of the College of Veterinary Medicine and Center for Feline Health and
Wellbeing Michigan State University
Venturing into the unknown: characterization of the aerobic cultivable bacteria in wounds of wildlife species
Courtney Pike, Lois Hoyer, Carol Maddox, Matthew Allender, Julia Whittington
Traumatic injuries, including blunt-force trauma and bites, are common in wildlife species. A lack of data detailing the
microbiota of wildlife wounds, as well as unknown history and limited resources, hamper treatment of these injuries. The
goal of this work was to characterize the aerobic cultivable bacteria from wounded animals brought to a wildlife hospital
during summer 2016. We hypothesized that the bacterial populations in wounds and on the peripheral skin would be similar.
Our secondary hypothesis was that wounds caused by contaminated sources would contain additional bacteria not found on
the skin. Upon admission, open wounds and the peripheral skin of wildlife species were individually sampled with a double-tipped swab, taking care to avoid cross-contamination. One swab from each site was used to plate the sample on blood,
MacConkey, and Columbia Naladixic Acid agar, then incubated for 16 hours. The second swab was used for Gram staining.
Pure cultures were derived for further identification. Isolates were analyzed using Matrix Assisted Laser Desorption Ionization-Time of Flight (MALDI-TOF) mass spectrometry, which is a newly emerging, cost-effective, and time-saving method
in diagnostic microbiology. Genus-level identifications were accurately achieved and deemed clinically sufficient for this
study. Preliminary data suggested that the MALDI-TOF is an attractive and inexpensive option for the wildlife veterinarian.
Understanding the normal bacterial populations on the skin of wildlife species provides insight regarding bacteria that may
be present in a wound and facilitates judicious antimicrobial use.
Research Grant: University of Illinois College of Veterinary Medicine
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Optimal decellularization produces synovial extracellular matrix scaffold for in vivo cartilage regeneration
Erin F. Pinnell, Logan M. Scheuermann, Nathalie A. Reisbig, Hayam A. Hussein, Alicia L. Bertone
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH
Decellularized extracellular matrices [ECM] can serve as biologic structural scaffolds for tissue regeneration. Prior work
in our laboratory [Reisbig, Pinnell, Hussein, Bertone, In Press, AJVR, 2016*] produced a novel synovial origin ECM (synECM) meeting specifications for transplantation of low DNA and cellularity while maintaining tissue integrity. The objective of our current study was to optimize cell migration and engraftment in various scaffold sizes designed for application in
animal knee models. Synovial tissue was collected and decellularization was performed using 0.1% peracetic acid (PAA) as
published* with either magnetic agitation or shake agitator for 6 hours, repeated twice. DNA content and fragment size, cell
viability, and histology were evaluated on the synECMs. Subsequently, 6mm and 2mm synECM scaffolds were positioned
in 6mm inserts of co-culture wells (0.4mm pore size) in a medium with a 30% fetal bovine serum gradient. Viable synoviocytes were seeded onto the 6mm and 2mm synECMs in quantities of 0.5, 1, or 2 x 106 cells. After 5 days of culture histology, flow cytometry, and trypan blue staining were performed to assess distribution, cell number and viability of engrafted
synovial cells on the synECM. Objective outcomes will be compared between methods by Students-t test with a significant
level of P<0.05. Both techniques produced synECMs with low DNA content and no DNA fragments and the histology for
tissue integrity is still pending. Outcome assessments for the cell seeding experiment is ongoing. The impact of our work
is anticipated to produce consistent large numbers of living synECM grafts that could be engineered and implanted in vivo
adjacent to cartilage defects to promote regeneration.
Research Grant: College of Veterinary Medicine Research Council Grant
Student Support: Wolfe Research Fellow Grant
Elena Pires, Neelam Sharma, David Maranon, Douglas Thamm, Claudia Wiese
Department of Environmental and Radiological Health Sciences, College of Veterinary Medicine and Biomedical
Sciences (Pires, Sharma, Maranon, Wiese) and Flint Animal Cancer Center, Veterinary Teaching Hospital (Thamm),
DNA double-strand breaks (DSBs) are considered to be the most detrimental DNA lesions induced by ionizing radiation
(IR), radio-chemicals, and other damaging agents. Cells respond to DSBs by activating DNA damage response pathways
that induce DNA damage repair. Flawless DNA repair is essential for correcting DSBs, maintaining genome integrity, and
preventing cancer. One vital DNA repair pathway for mending DSBs is homologous recombination (HR) DNA repair.
Defects in HR lead to cancer in humans and dogs, although the role of HR for tumor suppression in dogs remains to be
elucidated. Moreover, the mechanism by which molecular players function in the HR reaction is still poorly understood.
In this study, we monitored the level of ongoing HR DNA repair in a large set of canine cancer cell lines through two
surrogate markers of DSB repair, gH2AX and RAD51, by immunofluorescence. Our preliminary findings demonstrate
that these markers work successfully in dog cells, and their staining pattern exhibits a time and dose dependency. We also
characterized the DNA-binding properties of recombinant human RAD51AP1, a protein that is critical for proper HR and
for enhancing RAD51 activity in HR. Using the immobilized template assay, we find that full-length RAD51AP1 avidly
associates with both naked and chromatinized double-stranded (ds)DNA. To further define its DNA- and chromatin-binding
regions, we divided the protein into three fragments: F1, F2, and F3. Based on our findings with these fragments and other
results, we propose a model on the function of RAD51AP1 in HR. Taken together, the outcomes of our investigation will
help improve clinical treatment strategies in mammalian cancer patients with HR DNA repair defects.
Research Grant: NIH R01 ES021454; 2016 Young Investigator Grant Program, Center for Companion Animal Studies,
Colorado State University
Student Support: DVM/PhD graduate student stipend, College of Veterinary Medicine and Biomedical Sciences, CSU
233
Tyler Poole, Cheryl Jones, and S. Mark Tompkins
Department of Infectious Diseases, College of Veterinary Medicine, University of Georgia, Athens, GA
Ferrets are considered the gold standard for study of influenza infection, but are not fully utilized due to the lack of immune
reagents such as antibodies and cytokine proteins. Therefore, we have designed and cloned ferret cDNAs and show them
to express predicted full-length protein sequences for four ferret cytokines (l1, l3, b,and g). Here we are testing available
cell culture systems for optimal recombinant protein expression and purification. Synthesized ferret cDNAs for these proteins were cloned into the expression plasmid pcDNA 3.1 (-) A myc-His and transformed into bacteria JM109 for plasmid
DNA production. Since these vectors include a 6x Histidine tag and are secreted, the recombinant proteins can be purified
from cell culture supernatants using nickel column chromatography. Plasmid DNA was purified and the cDNA inserts
were confirmed by restriction digest and gel electrophoresis. The plasmids (pdDNA3.1- l1, pdDNA3.1- l3, pdDNA3.1- b,
and pdDNA3.1- g) were then used in transfections to assay protein expression. High-efficiency transfection systems were
used to express proteins in HEK-293 Freestyle and Expi-CHO cells. The Freestyle 293 cells were transfected using Freestyle Max or PEI as transfection reagents; the CHO cells were transfected using Epifectamine. The cells and supernatants
were collected over a time course and tested for protein production by western blot and Immunohistochemistry. Optimum
expression conditions will be used for scale-up. Purified proteins will be confirmed for biological activity and as antigens
for monoclonal antibody production. Ultimately these cytokines and antibodies will significantly improve the utility of the
ferret model for influenza and other research.
Research Grant: This work was funded in part by the NIAID Centers of Excellence in Influenza Research and
Surveillance (CEIRS), contract number HHSN272201400004C (to SMT).
Environmental Reservoirs of Francisella tularensis: Mechanisms for Transmission of Tularemia
Stephanie M. Porter, and Richard A. Bowen
Department of Biomedical Sciences, Colorado State University, Fort Collins, CO
Francisella tularensis is the causative agent of the zoonotic disease tularemia. Numerous mammalian species are susceptible to infection, and disease is commonly diagnosed in wild animals such as lagomorphs and various rodents. In susceptible
species, infection with type A strains typically results in septicemia and is frequently fatal without antibiotic treatment.
Transmission can occur by ingestion, inhalation, exposure of mucous membranes, and via arthropod-bite, but importantly,
a prominent unanswered question with respect to tularemia is how the bacterium is deposited in and survives in the environment, and is subsequently transmitted to mammals. Our objective was to obtain a more detailed understanding of the
dynamics of transmission of F. tularensis among natural hosts living in an ecologically relevant environment. In order to
do so, we characterized transmission of field isolates F. tularensis among wild mice (Peromyscus nasutus) housed in an
artificial ecosystem. Mice from the genus Peromyscus have been implicated previously as natural hosts of F. tularensis, and
cannibalism has been documented as a mode of transmission in an experimental setting.
Research Grant: Animal Models Core CSU
Student Support: USDA-NIFA Animal Health & Disease Research Program Funding 2016-36100-06008
234
Jonathan L. Powers, William H. Witola
Department of Pathobiology, College of Veterinary medicine, University of Illinois Urbana-Champaign, Urbana, IL
Toxoplasma gondii is an obligate, intracellular, zoonotic, parasitic protozoa that infects about one third of the world population. There are no drugs nor effective vaccines to control T. gondii in livestock. In humans, current drugs used against T.
gondii are limited by hypersensitivity and toxicity, and are only effective against the tachyzoite stage of T. gondii. Treatment
is necessary in immunocompromised patients and infants infected congenitally who may experience severe symptoms such
as eye damage, neurological pathologies, and difficulty breathing. The main goal of this study is to evaluate the efficacy of
green algae extracts in killing T. gondii proliferative-tachyzoite stages in vitro. Algae has been shown to possess antibacterial and antifungal properties, however, no studies have been performed to test whether it has any anti-protozoal effects.
We obtained a pure culture of green algae, Chlorophyceae, and an isolate of pond green algae. The samples were dried and
treated with either methanol or hexane. The methanol and hexane extracts were dried by evaporation and reconstituted in
DMSO. Confluent human foreskin fibroblasts were infected with T. gondii tachyzoites expressing YFP. Varying concentrations of the algae extracts were added and the cultures incubated for 48 h. Parasite growth was quantified by fluorescent
microscopy. We found that both the methanol and hexane extracts from the pure culture and pond algae had concentration-dependent anti-Toxoplasma effect at low microgram concentrations. We will derive anti-Toxoplasma IC50 values and
determine the cytotoxic IC50 values in HFF cells. Possible implications of this work may include a new treatment for a T.
gondii infection in humans and animals.
Research Grant: USDA Agriculture and Food Research Initiative
Nickel homeostasis and bacterial pathogenesis: Examining the NikR regulatory system in Brucella abortus
Evymarie Prado-Sanchez, James A. Budnick and Clayton C. Caswell
Blacksburg, VA
Brucella spp. are Gram-negative bacteria that infect a range of domestic and wild mammalian species causing abortions,
infertility and a debilitating febrile illness in humans. Some Brucella strains are considered zoonotic; therefore posing significant public health concern. As an intracellular pathogen, the bacteria invades phagocytes (i.e., macrophages, dendritic
cells, placental trophoblast), and compromise the regular activity of the infected cells. The overall goal of this study is to decipher the role of a metal responsive transcriptional regulatory protein called NikR in the pathogenesis of Brucella abortus.
Metals are essential micronutrients for the support of cellular metabolism and physiology of the brucellae, and in particular,
nickel is a co-factor required for the urease enzyme of Brucella. Overall, there is a need for novel therapeutic interventions
for both Brucella infections in mammals and humans and, in the end, it may be possible to target nickel homeostasis systems
to treat infections by Brucella. To accomplish our goal, we developed a plasmid that contains an in-frame deletion of nikR
for Brucella abortus. The plasmid will be introduced in the parental B. abortus 2308 strain and a variety of assays will be
performed to assess the role of nikR in nickel homeostasis and virulence of Brucella. Additionally, a recombinant Brucella
NikR protein (rNikR) was produced in E. coli BL21 and purified by affinity chromatography. Electrophoretic mobility shift
assays will be employed with different DNA promoter regions of nickel related operons to assess the DNA binding capacity
of the rNikR protein. These findings will serve to characterize the genetic regulatory events mediating nickel homeostasis
in Brucella.
Research Grant: Virginia-Maryland College of Veterinary Medicine
235
Characterization of the role of type I and type III interferons in acute and chronic disease
Emma Price, Yan Sun, and Carolina B. Lopez
Department of Pathobiology, University of Pennsylvania School of Veterinary Medicine
Respiratory viruses are major causes of hospitalization in humans and can cause chronic post-viral lung pathology such
as asthma, or even death. As protection against viral infections, humans and animals have evolved an extensive system to
detect foreign particles and trigger a rapid innate immune response. Within the nose, trachea, and lung, type I (IFN-I) and
type III interferons (IFN-III) are the first defense stimulated. However, the kinetics and distribution of IFN-I/III within
these respiratory compartments and their impact on viral clearance are still unclear. Furthermore, clinical studies suggest
that abnormally high or prolonged levels of IFN-I/III could trigger a malfunctioned state and thus contribute to disease
pathogenesis rather than having a protective role. Therefore, it is necessary to fully characterize the kinetics and roles of
both IFN-I and IFN-III during different time points and in different compartments of the respiratory tract during respiratory
viral infection. Here we use the murine parainfluenza virus Sendai virus (SeV) as a model to profile IFN expression in WT
mice and begin assessing their impact on tissue pathology. Also, using IFN-I receptor knockout (IFNAR KO) and IFN-III
receptor knockout (IL28aR KO) mice, we will investigate the impact of different IFNs on viral transmission and clearance.
Based on recent preliminary data, we hypothesize that at the acute phase of viral infection, IFN-III is primarily induced in
the upper respiratory tract while IFN-I is mostly expressed in the lung. Our study will provide a basis for the role of IFN-I/
III in different respiratory compartments during acute and chronic viral infection, leading to new targets for treatment and
prevention of lung disease.
Research Grant: NIH
Student Support: NIH and University of Pennsylvania Institute for Immunology
Examining the relationship between affective state and social rank in group-housed gestating sows
Katherine Pruett, Kristina Horback, Thomas Parsons
University of Pennsylvania School of Veterinary Medicine, Philadelphia PA
Societal concerns about the welfare of farm animals are driving initiatives to transition gestating sows from individual
stalls to group housing. In the absence of gestation stalls, animals have greater mobility but the establishment of the social
hierarchies can result in injuries and competition for resources. Common methods of measuring sow welfare tend to focus
on physical ailments such as lameness, poor body condition or skin lesions while neglecting measures of psychological
welfare. Cognitive bias testing is a novel tool researchers can use to assess the psychological welfare, or affective state, of
animals. There is evidence that animals experiencing negative affective states (such as anxiety or fear) are more likely to
interpret ambiguous information as threatening. Applying this information, 18 group-housed sows were trained a go/no-go
task using a positive rewarded stimulus and a negative punished stimulus. Each sow’s response toward an ambiguous stimulus was recorded as a measure of a sow’s affective state, and thus its welfare. This information, combined with feed order
hierarchical rank, intergroup fighting data, and body lesion scoring was used to determine how sow welfare is affected by
social rank in group pens. It is hypothesized that sows of lower rank would receive more aggression from other sows, have
more body lesions, and would be less likely to react positively to an ambiguous stimulus. Knowledge of individual differences among sows and hierarchies in group pens, and how this relates to coping and welfare, will be useful for improved
stockmanship and sow organization if and when group-housing is implemented.
Research Grant: Kraft Foods/ The Oscar Mayer Company
Student Support: NIH/Merial Summer Program, NIH T35 Training Grant
236
Maternal markers of inflammation and oxidative stress following air pollution exposure during pregnancy
Jairus Pulczinski, Kristal Rychlik, Natalie M. Johnson
Department of Environmental and Occupational Health, School of Public Health, Texas A&M University,
College Station, TX
Exposure to fine particulate (PM2.5) in utero has been shown to increase the risk of many diseases, including asthma in
offspring. However, the mechanistic link between maternal exposure and subsequent offspring asthma state remains unclear.
As inflammation and oxidative stress have been implicated in the skewing towards T Helper 2 dominate phenotype, a characteristic trait of allergic asthma, we used a One Health approach to examine the effects of PM2.5 exposure on 8-isoprostane and C-reactive protein, markers of oxidative stress and inflammation, respectively in both pregnant women and mice
exposed to air pollution in an effort to examine the conservation of air pollution induced inflammatory markers. Pregnant
women (n=15) PM2.5 exposure was quite variable with 24 hour averages ranging from 5 to 67 mg/m3. C-reactive protein
levels were consistently elevated across most participants, and ranged from 0.1 to 26.9 mg/L, with 80% of participants
above normal values. 8-isoprostane levels were generally lower than published normal values and ranged from 21.6 to
148.7 pg/ml. Among the rodent experiment model, mice exposed to air pollution had consistently lower levels of circulating
8-isoprostane, than their free air counterparts and ranged from 4.2 to 43.9 pg/ml. The human data failed to produce significant associations between exposure and plasma markers, likely a byproduct of the complex physiological changes occurring during pregnancy, as well as variation in personal habits, exposures, and baseline health status. Additional work will
examine the relationship of vitamin status, baseline health status, and other maternal factors to tease apart the contributors
to overall inflammation and oxidative stress.
Research Grant: Texas A&M School of Public Health REDI Texas A&M One Health On-Campus Summer Research Program
Student Support: Texas A&M One Health On-Campus Summer Research Program
Ashley K. Putman, Matt J. Kuhn, Jeffrey C. Gandy, and Lorraine M. Sordillo
Department of Large Animal Clinical Sciences, College of Veterinary Medicine, Michigan State University,
East Lansing, MI
Cytochrome P450 (CYP) molecules are oxidative catalysts that exert action on xenobiotics and endogenous compounds,
such as vitamin D. Cytochromes, along with cyclooxygenase and lipoxygenase, are able to metabolize polyunsaturated
fatty acids into oxylipids, which are potent inflammatory mediators. Depending on which cytochromes are expressed, metabolism of fatty acids through the CYP pathway can result in the biosynthesis of oxylipids with either anti-inflammatory
or pro-inflammatory activities. However, little is known about the expression of cytochromes in the bovine species. We
hypothesize that the basal expression of cytochromes will be differentially expressed in various bovine cells and tissues.
Bovine mammary and kidney epithelial cells and aortic endothelial cells were cultured in preparation for RNA extraction.
RNA will also be extracted from bovine kidney, lung, mammary, uterine, and liver tissues and then converted to cDNA.
RNA concentration and purity will be assessed using spectrophotometry and integrity analyzed using a bioanalyzer. qRTPCR will be used to amplify and quantify which cytochromes are expressed in the different cell and tissue types. The liver
expressed all genes that were assayed for, with the exception of CYP27B1. CYPs 2J2, 3A4, 4A11, and 24A1, along with
soluble epoxide hydrolase, were common amongst all tissue types. All cell types expressed CYPs 3A4, 24A1, and 27B1.
This study provided insight on the basal cytochrome P450 expression of various bovine cells and tissues. Future studies
directed towards the impact of inflammation on bovine cytochrome P450 expression are necessary to assess their potential
as novel targets during the treatment of inflammatory processes, such as mastitis.
Research Grant: MSU CVM Meadow Brook Laboratory
Student Support: NIH 4T35OD016477-14 grant
237
Karen Ramirez, Rhett Stout, Anderson DaCunha, Javier G. Nevarez
Louisiana State University, Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Baton Rouge, LA
70803
The objective of this study was to establish baseline electroencephalographic (EGG) parameters in reptiles when awake and
anesthetized. Eight adult, captive, box turtles (Terrapene carolina spp.) were tested. EEG needles were placed subcutaneously on the head. Two recording electrodes and one ground electrode were used. Turtles were administered alfaxalone at
20 mg/kg IM in the front legs. None of the turtles reached an anesthetized state with the dose of alfaxalone. Instead they
were sedated as evidenced by continuous voluntary movement of the limbs. There was however decreased tone of the head
and jaw tone and the animals did not appear to be responsive to visual stimuli. All animals had a characteristic posture of
extension and increased tone of the hind limbs with flaccid front limbs, head and neck. Awake and sedated readings were
obtained for all turtles. Analysis of the EEG’s is pending, however there was a visible decrease in the amplitude of the waves
in sedated versus awake state and a return to baseline upon recovery from sedation. By measuring the brain activity of these
species during these conscious and sedated states will help improve welfare and care of these animals in future anesthesia
and euthanasia studies.
Student Support: Department of Veterinary Clinical Sciences, LSU School of Veterinary Medicine
Biosensing of biofilm: detecting volatile organic compounds using a dog’s nose
Meghan T. Ramos, Thomas P. Schaer, and Cynthia M. Otto
Penn Vet Working Dog Center (Ramos, Otto), and Comparative Orthopedic Research Laboratory (Schaer),
University of Pennsylvania School of Veterinary Medicine, Philadelphia, PA
A biofilm is a microbe-derived sessile community characterized by cells that are attached to a substrate, or an indwelling
implant such as an orthopedic or cardiovascular device. Biofilm communities are covered with a “slime layer” composed of
a complex structure made of aggregates of microbial cells within a matrix of extracellular polymeric substances (EPS). Once
established, biofilms often cause persistent infection refractory to antimicrobial therapy. The release of toxins, inflammatory
mediators and streamers (planktonic bacterial cells) can further contribute to host morbidity. Biofilm infections are a diagnostic challenge for clinicians, resulting in increased healthcare costs, pain, and suffering of infected patients. Recent literature has demonstrated success in the utilization of medical detection dogs for scent detection of volatile organic compounds
(VOC) in human cancers. Based on the success of using dogs to detect the VOC of cancer tissues, we hypothesized that dogs
can identify the VOC biomarkers from Staphylococcus aureus biofilms. Three medical detection dogs were imprinted on
VOCs of cultured S. aureus. After the imprinting stage, the dogs systematically searched the scent detection wheel containing the S. aureus (positive) sample and appropriate negative controls. Once the dogs identified the positive sample within
the wheel they performed a trained indication. The three dogs demonstrated that they can detect the VOC biomarkers from
10^4 colony forming units of S. aureus with greater than 85% sensitivity and an 95% specificity over 5 trials.
Student Support: Supported by NIH grant T35 OD-010919 and a grant from Merial.
238
Evaluating body condition score with body weight on serum drug levels of phenobarbital and potassium bromide
Chelsea F. Randall, Devon W. Hague
Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois at Urbana-Champaign,
Urbana, IL.
Epilepsy is one of the most common neurological diseases affecting companion animals and seizures are managed through
the use of anticonvulsants. Anticonvulsants require regular therapeutic drug monitoring and medication adjustment by the
use of a first-order kinetics equation; however it is not always successful. The aim of this retrospective project was to assess
whether a patient’s body condition score (BCS) affects the serum level of phenobarbital and potassium bromide and determine if inclusion of this factor would result in an improved approach for medication adjustments. Data were collected from a
retrospective review of canine epilepsy patients at University of Illinois Veterinary Teaching Hospital from January 1, 2000
to June 1, 2016. Records that reported a phenobarbital and/or potassium bromide dosage (mg/kg), serum level, and BCS
were included in the study. The data set currently includes 113 dogs on phenobarbital, 29 dogs on potassium bromide, and 27
dogs on both therapies. Once the data set is complete, statistical analysis will assess whether BCS affects serum drug levels.
A novel drug adjustment approach using BCS could transform epilepsy treatment and increase the patient’s quality of life.
Research Grant: None.
Student Support: Merial Veterinary Scholars Program.
Environmental awareness: Recreating the natural tissue niche in vitro for cancer research
Brittany Rasche, Shirisha Chittiboyina, Farzaneh Atrian, and Sophie A. Lelievre
Department of Basic Medical Sciences, College of Veterinary Medicine, Purdue University, West Lafayette, IN
Despite many advances in cancer research, in vitro models that accurately represent the environment in which cancer develops have not been established. One example of the environment’s role is the link between increased stromal density and
increased risk for onset and progression of aggressive forms of breast cancer. Our hypothesis is that stromal stiffness and
cells affect the phenotypes of non-neoplastic and preinvasive mammary epithelial cells by influencing the morphology of the
cell nucleus. In a first approach, we are building a model that includes cancer cells, non-neoplastic epithelial cells, basement
membrane component (laminin), and stromal components (collagen I and fibroblasts). First, non-neoplastic and preinvasive
cells were cultured separately in the presence of collagen I matrix of adjustable stiffness. The nuclear morphology was
assessed based on DAPI staining (for nuclear areas and circularity) and SC35 immunostaining (for normal differentiation)
using ImageJ. The non-neoplastic cells showed a significant difference in nuclear area and circularity as well as differentiation, while the preinvasive cells displayed a significant difference in nuclear circularity depending on stiffness. Next, the
non-neoplastic and preinvasive cells were cocultured on top of the collagen I matrix. A significant alteration in nuclear area
for preinvasive cells was measured as stiffness changed. In future experiments, fibroblasts will be embedded in the collagen
matrix underneath the coculture of epithelial cells. The data collected from these experiments will be compared with data
from real breast tissue to determine which local environment allows cells to establish a phenotype that most closely resembles the real tissue.
Research Grant: Trask Innovation Fund to SAL
239
Morgan H. Rash, Karanvir S. Aulakh, Katherine H. Barnes, and Harmeet K. Aulakh
Louisiana State University, Veterinary Clinical Sciences, School of Veterinary Medicine, Baton Rouge, LA
Surgical resection of the damaged portion of the meniscus, while leaving as much normal tissue intact as possible, is recommended in dogs with meniscal tears. This procedure is associated with poor long-term outcomes in dogs. In humans,
meniscal transplantation is frequently performed with good outcome. However, studies have shown that improperly size
matched grafts have a much higher rate of failure, and the estimated tolerable margin of error is only about 10%. In veterinary literature, information regarding the utility of allograft transplantation is sparse. Consistently obtaining fresh graft material is difficult, necessitating the development of preservation and storage methods for the menisci. Cryopreservation has
been shown to have little to no effect on the morphology or biochemical properties of canine meniscus, however, effects of
cryopreservation on meniscal size has not been evaluated. This study aims to examine the effect of cryopreservation on the
size of the canine meniscus. We hypothesized that the size of the menisci will decrease with increasing duration of storage
time until reaching a fraction of their original size at which there will be no further decrease. Forty-eight menisci were taken
from a total of 14 canine cadavers. The hind limbs were disarticulated at the stifle joint and the size of each meniscus was
measured before being removed from the tibia. Once removed from the tibia, the measurements were repeated. All measurements were taken using a pair of Starrett digital calipers. The menisci were then preserved using a previously described
cryopreservation technique. Twenty-four menisci,12 lateral and 12 medial, were thawed and measured at 3 and 6 weeks.
Their size was recorded and compared.
Research Grant: Summer Scholar Grant, LSU
Student Support: Louisiana State University School of Veterinary Medicine
Structural modifications to K+ channel, Kv1.1, in a wild rodent resistant to scorpion venom
N Eden Reinhard, Kaitlyn R Sherer, Ashlee H Rowe
College of Veterinary Medicine (Reinhard, Sherer), Neuroscience Program (Sherer, Rowe), and Department
of Integrative Biology (Rowe), Michigan State University, East Lansing, MI
The Arizona Bark Scorpion (Centruroides sculpturatus) inflicts lethal and painful venomous stings to deter predators.
However, a species of grasshopper mice (Onychomys torridus), carnivorous rodents native to the southwestern US, prey on
these arthropods and are resistant to the painful and lethal effects. A more northern species, Onychomys arenicola, would
be killed if it tried to prey upon the Arizona Bark Scorpion and got stung, yet it feeds on the less toxic (but still potentially
lethal) Striped Bark Scorpion (Centruroides vittatus). Scorpion toxins are small peptides that interact with sodium (Na+)
and potassium (K+) channels which control neuronal firing. The toxins cause hyperexcitability by activating Na+ channels
to depolarize neurons, and by blocking K+ channels to prevent repolarization. Because the intensity of pain is directly correlated with the number of action potentials fired, scorpion toxins that more effectively cause excessive depolarization of
neurons in the pain pathway and inhibit K+ channel’s ability to control AP firing will cause more intense pain and damage.
Biologically, what allows O. torridus the ability to prey on more dangerous scorpions? We performed a combination of
reverse transcriptase polymerase chain reactions and Sanger sequencing, as well as cloning using TOPO-XL vectors. We
aligned the sequence of amino acids in the kcna1 (K+ channel) gene from dorsal root ganglion and trigeminal ganglion
to that of sensitive species to find sequence variation. One unique variant was observed in O. torridus, on the segment 4
voltage sensor, that was not found in O. arenicola. We hypothesize that this novel variant could play a role in the increased
resistance of O. torridus to scorpion toxin.
Research Grant: NSF IOS Award #1448393
240
Demonstrating C. botulinum neurotoxin heavy chain cytosolic localization via modified Kirby-Bauer assays
Abby Reising, Mengfei Ho, Jamie Perry, Melissa Pires-Alves, Ema Khan, Brenda A. Wilson
College of Veterinary Medicine (Reising) and Department of Microbiology, School of Molecular and Cellular Biology
(Ho, Perry, Pires-Alves, Khan, Wilson), University of Illinois at Urbana-Champaign, Urbana, IL
Clostridium botulinum produces the most potent toxins known to man, which inhibit acetylcholine release from neuronal
cells causing flaccid paralysis. The long half-life of the toxins in cells paired with the lack of an available antitoxin that
can act once the protein has entered the cell makes exposure untreatable and lethal after the critical prophylactic window.
This notion and the possibility of toxin use in bioterrorism makes it crucial to find a novel antitoxin. This study harnesses
the binding domain (BD) and the translocation domain (TD) of the heavy chain (HC) of botulism neurotoxin serotype A
(BoNT/A) to bind to neuronal cells, transfer from vesicles into the cytosol, and deliver therapeutic machinery necessary
to prevent further neurotransmitter inhibition. The heavy chain was linked to a prototype cargo, beta-lactamase (Bla), to
exemplify a means by which the cargo was successfully delivered to the cytosol with enzymatic activity intact. To test
for this delivery, modified Kirby-Bauer assays were developed. The positive control protein of Bla-LD-BD that utilized a
modified translocation domain (LD) and negative control protein of Bla-BD that lacks a TD or LD have been expressed,
purified, and assessed for their activity via Kirby-Bauer assays. Absence of a zone of clearing around an ampicillin-treated
disk, indicating the localization of beta-lactamase in the cytosol and the maintenance of its ability to cleave the beta-lactam
ring of ampicillin, reflects successful delivery of the Bla cargo. If efficacious, the HC of BoNT/A or the modified LD-BD
could be used not only as a vehicle for delivery of antitoxin cargo, but also as a means to deliver other new therapeutics to
neuronal cells.
Research Grant: National Institute of Allergy and Infectious Diseases, NIH, R33 AI101504
Rebecca M. Rettkowski, Luis E. Escobar, Alyssa A. Gohr, Nicholas B.D. Phelps
College of Veterinary Medicine (Rettkowski, Gohr), University of Minnesota, Saint Paul, MN; Minnesota Aquatic
Invasive Species Research Center (Escobar, Phelps), University of Minnesota, Saint Paul, MN
Fish are ecologically and economically important to the State of Minnesota. They are a source of food, provide recreational
value, and studying them helps us to better understand the health of our environment. Fish kills - defined here as a die-off
of more than five fish with similar signs, in close proximity, and over a short period of time - are, unfortunately, common
events in Minnesota, but limited research has been conducted to determine the causes of these kills. Within the last four
years, reporting systems and protocols have been significantly updated to better record and investigate fish kills across the
state. Due to these advances in reporting, recording, and organizing the information from the kills, a risk map was able to be
developed. It was hypothesized that fish kills in Minnesota are randomly occurring events in space and time. Using QGIS,
ArcGIS, and Maxent technologies, remote sensing variables explaining land use, climate, bathymetry, and human density
were analyzed, and their relationships to fish kills were determined. It was found that the maximum and average night temperature, cropland, and human density were all highly associated with fish kills, suggesting that climate and anthropogenic
disturbances could be linked to this phenomenon. A risk map of fish kills in Minnesota was developed to visualize this
analysis to allow future conservation officials to better understand the conditions associated with fish kills and to guide mitigation and surveillance efforts to prevent future fish kills.
Student Support: Funding was provided by Merial Limited and the College of Veterinary Medicine, University of MN
241
Mast cells in allergies and in asthma- does particulate matter (PM) activate mast cells?
Deja Rice, Christopher Occhiuto, Dennis Shubitowski, and Hariharan Subramanian
Department of Physiology, Michigan State University, East Lansing, MI-48820
Allergic disease are among the major cause of illness and disability in the United States. About 30 million Americans
suffer from allergic asthma and billions of dollars are being spent annually for hospitalization and treatment of asthmatic
patients. Therefore, there is an urgent need to understand the pathophysiology of these diseases in order to develop novel
therapeutics. Mast cells (MC) are immune cells that are critical mediators of allergic diseases. Activation of MC cause the
release of granules (via degranulation) that contain inflammatory mediators that result in symptoms associated with allergic
diseases. While allergies are specifically caused by allergens, it has been reported that environmental pollutants such as
diesel exhaust particles (PM) can amplify allergic diseases. However, it is unknown if PM can induce allergy via mast cell
activation. Thus the major goal of the study was to determine if PM activates human and mouse MC in vitro. Both human
and mouse MC were stimulated with PM obtained from different sources. (Detroit and Beijing). MCs were exposed to different concentrations of PM for 30 minutes. MC degranulation was assessed calorimetrically by estimating the release of the
enzyme b-Hexoseaminidase. We used the complement component C3a and IgE/antigen as our positive control. While C3a
and IgE/antigen induced a robust degranulation response in human and mouse MC (~40%) respectively, PM from Detroit
and Beijing did not induce degranulation in MC (<10%). These data indicate that PM do not activate MC. However PM
may synergize with other MC activation pathways (for e.g.,IgE/antigen) to amplify allergic disease. This possibility will be
investigated in the future.
Active veterinary surveillance of emerging Ixodes scapularis tick populations in Michigan
Meredith K. Rice, Megan Porter, Jean I. Tsao
College of Veterinary Medicine, Lincoln Memorial University, Harrogate, TN (Rice); College of Veterinary Medicine
(Porter) and Dept. of Large Animal Clinical Sciences (Tsao), Michigan State University, East Lansing, MI
Ixodes scapularis is the vector for Borrelia burgdorferi, which causes Lyme disease in humans and dogs. These ticks are
established in western Michigan and appear to be spreading eastward across the state. Understanding the locations of emerging tick populations will aid in the prevention of tick-borne disease. Because companion canines often encounter the same
environment as humans and are good hosts for blacklegged ticks, we developed a network of veterinarians in Michigan
to actively survey the state with broader coverage than prior efforts. Veterinary clinics and animal shelters were chosen at
random and were asked to submit ticks and information on each dog’s travel history, use of tick preventatives, and status of
Lyme disease vaccination during routine check-ups during Spring 2016. After mapping the zip codes of veterinary-submitted ticks, there was evidence of emerging populations in eastern Michigan. Several locations were chosen for field sampling
to investigate the presence of local, established ticks populations. To assess if these possibly emerging tick populations pose
disease risk, we assayed ticks for B. burgdorferi. From this study, we can conclude that veterinary surveillance can identify
potentially emerging blacklegged tick populations, which can help educate veterinarians and public health workers regarding the local risk of Lyme disease and target public health measures more efficiently.
Research Grant: NIH Grant No. T35OD016477 to Michigan State University; CVM Endowed Companion Animal Fund
No. RT082792-JT
Student Support: NIH Grant No. T35OD016477 to Michigan State University
242
Ella Richardson and Brian B. Oakley
Consumption of poultry contaminated with Campylobacter jejuni is an important cause of bacterial gastroenteritis worldwide. Consequently, reducing cecal colonization by Campylobacter jejuni during poultry production could have profound
food safety benefits. The use of subtherapeutic levels of antibiotics in animal feed has typically been used to modulate gut
pathogens and promote animal growth, but has led to the emergence of antibiotic resistance. One promising alternative to
the use of antibiotics in food animal production is the use of probiotic supplements isolated from naturally occurring gut
bacteria. Introducing pre-established bacterial communities to competitively exclude pathogenic bacteria is a strategy that
has been implemented in people receiving fecal microbiome transplants to treat refractory Clostridium difficile infections.
Similarly, we tested the efficacy of cecal microbiome transplants as a competitive exclusion strategy against Campylobacter
jejuni in the chicken gut. Using qPCR and fluorescence in-situ hybridization, we aim to quantify Campylobacter jejuni in
the cecal contents of chickens treated with and without cecal microbiome transplants.
Research Grant: USPOULTRY Foundation and College of Veterinary Medicine intramural funds
Student Support: CVM Research Scholars Program and Western U Summer Research Fellowship
Disparate virulence in two plaque variants of Theiler’s murine encephalitis virus, DA strain
Alesha Rimmelin, Megha Bijalwan, C. Jane Welsh, Colin Young, Ishita Bansal, Julian Leibowitz, and Judy Pham
Department of Veterinary Integrative Biosciences, College of Veterinary Medicine (Rimmelin, Bijalwan, Welsh, Young,
Bansal), and Department of Microbial Pathogenesis and Immunology, Health Science Center (Leibowitz, Pham), Texas
A&M University, College Station, TX
Theiler’s Murine Encephalitis Virus (TMEV) is a neurotropic single stranded RNA cardiovirus of the Picornoviridae family. It is useful as a model for epilepsy, progressive paralysis, encephalomyelitis, and demyelination. A variety of TMEV
strains exist, with different phenotypic expressions of virulence in different strains of mice. The DA strain of TMEV has
been shown to induce epileptic seizures in C57BL/6 mice. Two plaque variants of the DA strain, C and D, have previously
been shown to have differences in virulence, with C being markedly less virulent than D. In this study, these two plaque
variants were intracranially injected into C57BL/6 mice. Mice were observed twice daily and terminated at seven days post
infection. Seizure incidence, weight loss, and viral titers in the brain were significantly higher in the D-infected mice. Viral
titer in the brain correlated positively with seizure incidence, while viral titers in the spinal cord correlated negatively with
incidence of seizures. Brain weights of D-infected mice at termination were also significantly higher. Histological examination of brain sections showed the highest degree of perivascular cuffing, mononuclear infiltration, and pyramidal neuron
damage in the hippocampus of D-infected mice. In conclusion, the two plaque variants are demonstrably different in virulence which corresponds to an array of phenotypic changes in systemic and CNS health. Higher virulence correlated with
increased seizure rate and severity, viral replication in the CNS, microglial and macrophage activation, inflammation, and
neuronal damage. Currently, sequence comparisons of the two plaque variants are underway to characterize the molecular
differences between the plaque variants.
Research Grant: 1R21AI121807 (Leibowitz)
243
Endothelial colony forming cells as treatment for equine distal limb wounds
Rachel L. Roberson, Randolph L. Winter, Yuan Tian, Wen J. Seeto, Ashley N. Sharpe, Kelly A. Himeback,
Fred J. Caldwell, Elizabeth A. Lipke, Anne A. Wooldridge
Department of Clinical Sciences, College of Veterinary Medicine (Roberson, Winter, Sharpe, Himeback, Caldwell,
Wooldridge) and Department of Chemical Engineering, Samuel Ginn College of Engineering (Tian, Seeto, Lipke),
Auburn University, Auburn, AL
Endothelial colony forming cells (ECFCs) are progenitor cells that function in vascular repair and neovascularization.
Therefore, ECFCs may be useful in the treatment of conditions characterized by poor blood supply. An equine distal limb
wound model was used to evaluate the effects of treatment with ECFCs on wound surface area and granulation tissue formation. To improve cell survival and localization upon injection, ECFCs were also encapsulated in polyethylene glycol-fibrinogen (PEG-fibrinogen). Three horses had two 6.25 cm2 dermal wounds created on each distal limb. Each wound randomly
received 1 of 4 treatments by subcutaneous injection: serum, PEG-fibrinogen microspheres (MS), ECFCs, or ECFCS encapsulated in PEG-fibrinogen microspheres (ECFC-MS). Wound healing was assessed weekly using digital image wound
surface area (WSA) analysis and granulation tissue scores (GS) assigned by blinded observers. Four wounds per horse (1 per
treatment) were biopsied at baseline and then weekly while the other four wounds per horse (1 per treatment) were biopsied
at baseline and week 4 only. The effect of treatment group on the percent change in WSA was significant (p=0.0002), with
ECFCs alone having the smallest WSA measurements. The ECFCs alone and ECFC-MS groups had significantly smaller
WSA compared to MS alone (p=0.0005 and p=0.0014, respectively). Compared to serum, ECFCs alone had a WSA decrease which trended toward significance (p=0.0742). GS were greater for hindlimb wounds (p=<0.0001) and for wounds
biopsied weekly (p=0.0039). GS were also different due to the individual horse (p=<0.0001). Injection with ECFCs alone
or ECFC-MS significantly reduced wound surface area, which may indicate enhanced healing.
Research Grant: Grayson Jockey Club Research Foundation
Cassandra Rodenbaugh, Andrew Hanzlicek, Ian Kanda, Mark Payton, Shane Lyon, Theresa Rizzi, and Joao Brandao
Department of Veterinary Clinical Sciences (Rodenbaugh, Hanzlicek, Kanda, Lyon, Brandao) and Department
of Veterinary Pathobiology (Rizzi), Center for Veterinary Health Sciences, Oklahoma State University, Stillwater, OK.
Department of Statistics (Payton), Oklahoma State University, Stillwater, OK.
Avian blood takes longer to clot than mammalian blood, due to differences in circulating cells and clotting factors. These
differences make currently available mammalian coagulation assays of limited use in birds. In pursuit of a new, alternative
methodology, we used a Sonoclot Coagulation and Platelet Function Analyzer to measure the coagulation time of blood
samples from 30 privately owned hens. Using a balanced incomplete block design, we compared the coagulation profile
(fibrin formation and clotting rate) of fresh blood and sodium citrated blood, while stimulating coagulation with 1 of 3 activators: glass beads, tissue factor, or kaolin clay. Citrated samples were associated with an increased time until initial fibrin
formation, with glass bead-stimulated samples taking four times longer than fresh samples, and tissue factor and kaolin
samples taking twice as long as fresh samples. Citrated samples had decreased clotting rates of approximately 30% (kaolin),
40% (tissue factor), and 90% (glass beads) when compared to fresh blood with the same activator. Results of this study
suggest that sodium citrate leads to relative hypocoagulability of avian blood when compared with fresh blood. Although
commonly used as an anticoagulant for mammalian coagulation assessment, the results of this study indicate that sodium
citrate may not be optimal for chickens. Further research is necessary to determine whether the hypocoagulability associated
with the citrated samples is a result of physiological or in vitro interactions between sodium citrate and avian blood, which
are not observed with mammalian samples.
Research Grant: Oklahoma State University Center for Veterinary Health Sciences and Joan Kirkpatrick Chair in Small
Animal Medicine
244
Mutant XYZ is associated with hyper-activation of mTOR and T cell senescence
Hailey Rose, Jill Fritz, and Michael Lenardo
Molecular Development of the Immune System Section, Laboratory of Immunology, National Institute of Allergy and
Infectious Diseases, National Institutes of Health, Bethesda, Maryland
A 14 year old female Turkish patient from a consanguineous family has a clinical phenotype of reactive airway disease,
multiple lymphadenopathies, hemolytic anemia, low IgA and undetectable IgE, and positive antinuclear antibodies. Using
whole exome sequencing, a homozygous missense mutation in gene XYZ was identified in the patient and predicted to be
possibly damaging and disease-causing. XYZ is a kinase that participates in the biogenesis of phosphatidylinositol 4,5-bisphosphate (PIP2), an important signaling molecule involved in T cell activation. The mutation results in a tryptophan to
arginine amino acid change at position 410, which is located in the activation loop of the C-terminus. The patient T cells
have lower XYZ protein levels compared to controls. Further analysis of peripheral blood mononuclear cells in the patient
revealed that the frequency of memory B cells was severely reduced in the patient in association with low levels of classswitched immunoglobulins. The patient also had fewer naive T cells and increased senescent effector T cells. These findings
were further supported by a higher frequency of T cells that expressed CD57, a marker of T senescence. The patient also
exhibited reduced cell death in response to restimulation of the T cell receptor, a process that relies on active cell cycle entry.
Patient T cells stimulated with anti-CD3 had increased phosphorylation of S6, a signaling molecule downstream of mTOR.
Overall these findings suggest that the mutation in XYZ leads to hyper-activation of mTOR, thereby driving increased terminal differentiation and senescence in T cells.
Student Support: Comparative Biomedical Scientist Training Program
Loss of parvalbumin-immunoreactive interneurons in epileptic California sea lions
Tierra Rose, Starr Cameron, Raisa Glabman, Emily Abrams, Shawn Johnson, Frances Gulland, Paul Buckmaster
Purdue University, College of Veterinary Medicine (Rose), West Lafayette, IN. Stanford University, School of Medicine,
Department of Comparative Medicine (Cameron, Abrams, Buckmaster), Stanford, CA. University of Pennsylvania,
School of Veterinary Medicine (Glabman), Philadelphia, PA. Marine Mammal Center (Johnson, Gulland), Sausalito, CA
Temporal lobe epilepsy is common in humans. Seizures typically originate in the hippocampus, but the cause is unknown.
The hippocampal dentate gyrus of human patients displays neuropathological abnormalities, including the loss of parvalbumin-immunoreactive interneurons. Normally, parvalbumin interneurons strongly inhibit excitatory neurons. Loss of
parvalbumin interneurons in temporal lobe epilepsy might cause seizures. Rodent models of temporal lobe epilepsy fail to
replicate the parvalbumin interneuron loss found in human patients, so better animal models are needed. California sea lions
(Zalophus californianus) develop temporal lobe epilepsy after exposure to the excitatory neurotoxin domoic acid, which
enters the marine food chain during harmful algal blooms. We hypothesized that epileptic sea lions would display significant loss of parvalbumin interneurons in the dentate gyrus. To test this hypothesis, sea lions were intracardially perfused
with formaldehyde immediately upon euthanasia because of failed response to treatment and poor prognosis. Brains were
sectioned (40 mm) coronally. Hippocampi were isolated and processed for parvalbumin-immunocytochemistry. Stereology
and a Neurolucida system are being used to estimate the number of parvalbumin-positive interneurons per dentate gyrus.
Preliminary data suggest that epileptic sea lions display severe loss of parvalbumin interneurons, similar to human patients
with temporal lobe epilepsy. If confirmed, these findings would suggest that epileptic sea lions can be used as a large animal model of human temporal lobe epilepsy. Sea lions could serve as candidates to test novel anti-epileptogenic treatments
before human clinical trials.
Research Grant: NSF and NIH (NINDS & OD)
245
Parkinson disease modeling in parkin-deficient mice: testing synthetic mitochondrial-targeted antioxidants
Brandi Roseman
College of Veterinary Medicine, Tuskegee University, Tuskegee, AL. Merial Veterinary Scholars Program,
Auburn University College of Veterinary Medicine
Parkinson’s disease (PD) is the second most common neurodegenerative disease after Alzheimer’s disease. The etiology of
PD is complex, usually with a combination of both genetic and environmental factors. Previous studies have shown environmental hazards such as herbicides/pesticides can induce PD symptoms. For modeling PD in rodents, a pesticide called rotenone, is used for specific inhibition of complex 1 of the electron transport chain, causing oxidative stress and mitochondrial
dysfunction. Genetic factors include mutations in a protein called parkin, which is encoded by the PARK2 gene, leading to
defects in mitochondrial recycling and cell death by the intrinsic apoptotic pathway. PD is characterized by degeneration
of dopaminergic neurons in a particular part of the brain (called the Substantia Nigra pars compacta). Diminished levels
of dopamine in the brains of PD patients affects regulation of movements and emotions. In this study we tested synthetic
mitochondrial-targeted butyrylcarnitine compound (PMX550DBr) in the treatment of rotenone-induced mitochondrial dysfunction in a transgenic mouse model of PD. Intra-peritoneal (IP) injections of rotenone induced PD like symptoms in mice
harboring a PARK2 gene mutation. The mice were divided into 3 groups (control, rotenone only, and rotenone+ PMX550DBr) and assessed by neuromotor/behavioral testing and western blot from tissue samples to test whether co-treatment of
rotenone-treated mice with, PMX550DBr, protected mitochondria and improved the PD- like phenotype.
Research Grant: The MitoCure Foundation
Student Support: Merial & Auburn University College of Veterinary Medicine
Investigating the role of Delta-like ligand 1, a Notch signaling ligand, in breast cancer metastasis
Taylor Ross and Rumela Chakrabarti
Breast cancer is a major health threat and the second leading cause of cancer related death in women in the United States.
Notch signaling is shown to regulate both normal development of mammary glands and breast cancer, but the mechanism
is not understood. Toxicity associated with targeting Notch signaling with GSI drugs is a big clinical problem. Most prior
studies focus on Notch receptors, not ligands. We looked at the Notch ligand Dll1, which has not been studied in the context
of breast cancer, in hopes of safer ligand based treatment options. Unpublished research from the Chakrabarti lab suggests a
role of Dll1 in tumor initiation. Correlation data for metastasis in patient datasets shows a subtype specific correlation. This
data led to the hypothesis that Dll1 may have a subtype specific function in breast cancer. Dll1+ and Dll1- cells were sorted
using FACS analysis and confirmed by qPCR in a basal breast cancer cell line (4T1). In vitro experimentation shows that
Dll1- 4T1 cells have a proliferative advantage, whereas Dll1+ 4T1 cells have a migratory advantage and higher cancer stem
cell like activity. Preliminary in vivo data supports these findings. Mice that received Dll1- cells showed an early advantage
in tumor growth, but over time, the Dll1+ tumors grew faster. These Dll1+ tumors have a larger population of myeloid
derived suppressor cells compared to Dll1- tumors, possibly explaining this change. The lungs, livers, and spleens of these
mice are being analyzed for metastasis. Also, shRNAs are being validated for knockdown in basal and luminal breast cancer cell lines for further functional experimentation. In parallel, in vitro drug treatment data suggests that this gene can be
modified epigenetically.
Research Grant: ACS-IRG grant from Abramson Cancer Center, K22-NCI grant and Start up
Student Support: NIH grant T35 OD-010919 and a grant from Merial
246
Involvement of increased CB1R activation in altered social play induced by developmental chlorpyrifos exposure
Nicole E. Rowbotham, Carole A. Nail, Jenna A. Mosier, Aubrey M. Lewis, Russell L. Carr
Center for Environmental Health Sciences, Department of Basic Sciences, College of Veterinary Medicine, Mississippi
State University, Mississippi State, MS
Childhood exposure to chlorpyrifos (CPF), an organophosphorus insecticide, results in negative long-term neurologic effects. We have previously reported that low-level developmental exposure of rats to CPF disrupts endocannabinoid (EC)
degradation through fatty acid amide hydrolase (FAAH) inhibition and leads to increased social play behavior once adolescence is reached. Increased activity in the EC system may be responsible for the observed altered behavior. Phosphorylation
of the cannabinoid receptor (CB1R) is an indicator of EC activation. This study compared the phosphorylation of the CB1R
in brain regions of control and treated rats immediately following behavioral testing. On postnatal day (PND) 10, rats were
exposed orally to either corn oil, 0.5 mg/kg CPF, 0.75 mg/kg CPF, 1.0 mg/kg CPF or 0.02 mg/kg PF-04457845 (a specific
FAAH inhibitor) daily for 7 days. On PND 36, social behavior was monitored and all treated groups spent more time playing than did controls. Western blotting was used to quantify the amount of CB1R and phosphorylated CB1R proteins in the
hippocampus, amygdala, prelimbic cortex, agranular insular area, and nucleus accumbens. There were no significant effects
of treatment on the amount of CB1R protein in any of the five brain regions. Increased CB1R phosphorylation occurred
in all brain regions following social play, but no effects of CPF treatment were observed. This suggests that increased EC
activation is not a causative factor in the increased social play observed in CPF treated rats. It remains unclear whether
developmental CPF exposure induces an alteration in EC tone or an alteration in other neurotransmitter systems that could
explain the observed changes in social behavior.
Research Grant: NIH R15 ES023162
Intestinal epithelial cells NF-kB regulates host response to ingestion of low doses of cadmium
John C. Rowe, Eunsoo Kim, Haley E. Steiner, Estelle Cormet-Boyaka, Prosper Boyaka
Department of Veterinary Biosciences, College of Veterinary Medicine, Columbus, OH
Minor disruptions to the homeostatic equilibrium of the intestinal tract can lead to Inflammatory Bowel Disease (IBD),
Crohn’s disease, and ulcerative colitis, which affect approximately 1.4 million people in the United States. Low levels of
cadmium are commonly found in water runoff and accumulation can occur in plants, seafood, and soft tissues of mammalians. This heavy metal is now listed 7th in the priority list of hazardous substances and is believed to promote inflammation.
This study explored the role of intestinal epithelial cells (IECs), and more specifically the canonical NF-kB pathway of these
cells, in host mucosal responses to repeated ingestion of cadmium. Control wild-type C57BL/6 and IKKbDIEC mice, which
lack IKKb in IECs, were maintained in conventional SPF housing (n=5 per group) and provided cadmium as CdCl2 (10 uM
or 2 ppm) in drinking water for 14 days. Analysis of total sIgA in fecal samples collected on days 0, 7, and 14 showed that
cadmium treatment reduces sIgA levels in both groups of mice. We also found that repeated ingestion of cadmium differentially affected the frequency of lymphocyte subsets in mesenteric lymph nodes (MLNs) of IKKbDIEC and control wildtype mice and increased percentage of B cells while reducing the percentage of T cells in IKKbDIEC mice. Furthermore,
cadmium treatment enhanced gut TGFb and TNFa mRNA responses to the bacterial product cholera toxin in IKKbDIEC
mice. Taken together, our data suggest that the canonical NF-kB in intestinal epithelial cells plays a key role in host response
to environmental pollutants and subsequent inflammatory status in the gastrointestinal tract.
Research Grant: NIH
Student Support: Ohio State SOLAR Research Fund
247
Ellen Russell, Veronica Ringel, A. Jane Duncan, Daniel Slade, Lera Brannan, James Boone, Yufeng Qin, Paul Wade,
Irving C. Allen
Department of Biomedical Science and Pathobiology (Allen, Ringel) Virginia-Maryland CVM (Russell) and Department
of Biochemistry, College of Agriculture and Life Science, (Duncan, Slade), Virginia Tech, Blacksburg, VA TECHLAB,
Blacksburg, VA (Boone, Brannan) National institute of Environmental Health Sciences (Qin, Wade)
When a NOD-like receptor (NLR) senses a pathogen associated molecular pattern (PAMP), it triggers the creation of an
inflammasome. Inflammasome activation results in pyroptosis and cleavage of pro-inflammatory cytokines proIL-18 and
proIL-1b to their active forms. Consequently, the absence or malfunction of an NLR in the gut has been shown to result in
dysbiosis. Clostridium difficile is a commensal bacterium of the gut that can cause mild to deadly cases of colitis in patients
with a compromised microbiome. There is significant evidence linking dysbiosis with chronic inflammation in the gut and
colon cancer. Prior work in our laboratory has revealed that mice lacking specific NLR inflammasomes are highly susceptible to experimental colitis and colitis-associated tumorigenesis. Here, we extend these findings and show that ASC-/- and
Nlrp1-/- mice have an outgrowth of Clostridium in their resting microbiome, implying that the absence of a specific NLR
Inflammasome may permit excessive growth of Clostridium species. To investigate these findings further, we utilized bone
marrow derived macrophages from mice lacking canonical or non-canonical inflammasome components and evaluated
the effects of inflammasome dysfunction following infection with a variety of C. difficile strains. Our results indicate that
mice lacking canonical inflammasome components have diminished immune responses to C. difficile. Together, these data
suggest that the absence of specific NLR family members may create a permissive niche for Clostridium species in the gut,
which may result in increased colitis pathogenesis and ultimately tumorigenesis. Future work will better resolve which NLR
is responsible for inflammasome response to C. difficile.
Research Grant: NIH/NIDDK K01-DK092355
Effect of litter type on survival of Tritrichomonas foetus in feline feces
Elizabeth L. Sablotny, Stephen H. Stauffer, and Jody L. Gookin
Tritrichomonas foetus is a protozoal cause of lifelong large bowel diarrhea in cats. The only treatment has a narrow safety
margin and resistance is common. Use of litter types capable of rapidly killing T. foetus could ameliorate fecal-oral spread of
the infection. The objective of this study is to develop a simulated litter box model of T. foetus-infected feline diarrhea and
use this model to test the effect of litter type on T. foetus survival. Simulated T. foetus infected feces was created using equal
weights of stools collected from four cat-owning households, homogenized with DPBS solution to form a loose consistency,
and inoculated with 106 feline T. foetus per gram feces. Equal-weighted aliquots of fecal homogenate were both buried and
placed on the surface of each litter type. Samples from the original fecal homogenate and each litter box specimen were
collected at predetermined times, subcultured in T. foetus media, and examined for live trophozoites using an inverted microscope. Simulation of diarrheic feces was accomplished using an average 1.31:1 (w:v) ratio of stool to DPBS solution to
generate an average fecal wet weight of 77.75%. There was high variability in the number of T. foetus-positive subcultures
obtained directly from the fecal homogenate, ranging from 0-100%. Successful experimentation revealed that, when buried
in sand, T. foetus-inoculated feces survived a maximum of 22 hours compared to feces tested with crystal litter, which did
not yield viable T. foetus at any time point. These results show promise for this model as a method to determine if certain litters are more effective at decreasing environmental survival of T. foetus in feces to potentially lessen the spread of infection.
Research Grant: NCVMF Support for Tritrichomonas foetus Research, Innovation, and Veterinary Education (STRIVE)
Student Support: NIH Grant #T35OD011070
248
Effects of the JAK-inhibitor AZD1480 on activated dorsal root ganglia and itch behavior in mice
Laura Sanabria-Ojeda, Tomoki Fukuyama, and Wolfgang Baumer
Department of Molecular Biomedical Sciences, College of Veterinary Medicine, North Carolina State University,
Raleigh, NC
Janus Kinase (JAK) inhibitors are therapeutics for cancer and inflammatory diseases, such as atopic dermatitis (AD), that
block the signaling of cytokines, like IL-31, involved in itch perception. AZD1480 is a JAK-inhibitor with similar inhibitory
profile as oclacitinib, a JAK-inhibitor licensed for canine AD. The objective of this study was to determine the exact mechanism by which different JAK-inhibitors, in particular AZD1480, provide such a quick onset of action in controlling the itch
response both in vitro and in vivo. In vitro, the dorsal root ganglia (DRG) of untreated female BALB/c mice were collected
and enzymatically dissociated. Fura-2 ratiometric calcium imaging was used to evaluate the itch response of DRG neurons
to IL-31, histamine, and chloroquine, after being treated with JAK-inhibitors. In vivo studies were performed with female
BALB/c mice that were treated orally with AZD1480, 30 minutes prior to an intradermal injection of IL-31, histamine,
and chloroquine. The mice’s scratching behavior was recorded for 30 minutes after stimulus injection. Pre-treatment with
AZD1480 resulted in significant reduction of the in vitro response of DRG neurons to IL-31, histamine, and chloroquine.
Mice treated with AZD1480 also showed significant reduction in scratching behavior after stimulus with IL-31, histamine,
and chloroquine by 60.1%, 80.7%, and 83.0%, respectively. In conclusion, besides reducing pruritus induced by IL-31 as
expected, AZD1480 lowered the reaction induced by histamine and chloroquine. These are pruritic mediators that are not
controlled through the JAK STAT pathway, suggesting that JAK-inhibitors might work through more than one pathway
involved in itch perception.
Student Support: NIH- T35 Interdisciplinary Biomedical Research Training Program and NIH Grant #T35OD011070
Behavioral Temperament of rhesus macaques (Macaca mulatta) and the association with gut microbiota
Yu Sato, Amir Ardeshir
School of Veterinary Medicine at UC Davis (Yu Sato), California National Primate Research Center (Amir Ardeshir)
The role of gut microbial communities in host health, such as immune response has been studied rigorously in the past.
However, the link of microbiota - gut - brain axis in host behavior and psychological health remains unclear. Here, we
investigate the link between the early life gut microbial communities and the semi-quantitative assessment of the nervous
temperament in rhesus macaques. Our goal is to determine whether the composition of gut microbiota is associated with
nervous temperament of infant macaques. We collected 198 mother - infant dyads from 7 different social groups from outdoor colony at the California National Primate Research Center. All infants were born in 2015, and they were 3 to 4 months
of age at the time of behavioral assessment and fecal collection. Infant behavior was scored, and categorized into different temperament scale by using number of standardized colony-wide behavioral tests, BioBehavioral Assessment. We are
currently characterizing microbial communities of two cohorts, most (n=20) and least “nervous” (n=20) infant macaques
using 16S rRNA gene amplicon sequencing. To test whether infant macaques that demonstrate more nervous personality
have distinct gut microbiota, we apply dimension reduction methods and cluster analysis techniques in R. This study has a
potential to understand the role of gut microbes in human behavior and health, as well as to develop probiotics for humans
to treat psychiatric illnesses.
249
Temperature as a confounding variable in oncolytic virotherapy for canine melanomas
Julia Saturno, Jacob van Vloten and Byram Bridle
Department of Pathobiology, Ontario Veterinary College, University of Guelph, Guelph ON
High-grade canine melanomas are almost uniformly fatal despite aggressive interventions. The current standards of care are
expensive and time consuming, making treatment an unfeasible option for many pet owners. We published a strategy to use
oncolytic viruses (OVs) as booster vaccines to synergize benefits between immuno- and oncolytic virotherapies and have
developed infrastructure to test this in both companion animal and human clinical trials. The purpose of this study is to identify an ideal OV platform for canine melanoma therapy. The oncolytic potential of Maraba virus (MG1), vesicular stomatitis
virus (VSV), Newcastle disease virus (NDV) and vaccinia virus (VV) was evaluated in the patient-derived canine melanoma cell lines ICCI25cl.5 and CML1. Madin-Darby Canine Kidney (MDCK) epithelial cells were included as non-malignant
controls. To simulate elevated body temperatures of canines compared to humans, account for induction of fevers in patients
treated with OVs, and recapitulate the conditions of tumours located in diverse anatomical locations, efficacy was assessed
across a range of temperatures. A high-throughput resazurin dye-based metabolic assay was used to assess the viability of
OV-treated cells. All of the viruses demonstrated differential killing with various degrees of cytotoxicity in melanoma but
not MDCK cells. At higher temperatures both MG1 and VSV lost oncolytic activity while NDV and VV remained efficacious. Through heat-adaptation, we were able to restore the replication potential of MG1 at elevated temperatures. Clinical
considerations include using OVs that are less oncolytic but retain efficacy over a broad range of temperatures or utilizing
heat-adaptation for temperature-sensitive viruses.
Research Grant: National Center of Excellence in Biotherapies for Cancer Treatment (BioCanRx), Ontario Veterinary
College Pet Trust Fund
Michael Schacher, Amy E. DeClue, Sandra M. Bechtel, and Juliana Amorim
University of Missouri CVM, Columbia, MO
Sepsis is the leading cause of death in people in Intensive Care Units and a leading cause of death overall in the US. Recently, new insights into the pathophysiology of sepsis have led to the realization that immunosuppression along with the traditionally implicated cytokine storm contributes to morbidity and mortality. Moreover, we have preliminary data indicating
that some dogs with critical illness develop an immunosuppressive phenotype similar to that of people with sepsis. GM-CSF
reverses a similar immunosuppressive phenotype observed in dogs with cancer and thus might be a candidate intervention
for reversing the immunosuppressive phenotype in critically ill dogs. We hypothesized that GM-CSF would improve phagocytic function, oxidative burst capacity, phagocytic expression of TLR4 and HLA-DR, and LPS-stimulated production of
cytokines in cells from dogs with critical illness-induced immunodysfunction. Critically ill dogs were identified through the
University of Missouri, Veterinary Health Center. Blood was obtained and incubated with PBS (control), GM-CSF 0.1nM,
or GM-CSF 1.0nM for 2hrs. Then, phagocytosis of opsonized FITC labeled E. coli, and E. coli-induced respiratory burst
function were assessed. Fluorescently labeled HLA-DR and TLR4 antibodies were used to assess cell surface expression of
pattern recognition receptors on monocytes. Analysis of phagocytosis, oxidative burst capacity, and HLA-DR/TLR expression were evaluated via flow cytometry. Analysis of lipopolysaccharide-induced leukocyte TNF and IL-10 production will
be performed using a canine specific multiplex cytokine assay. At this time, blood has been collected from 4 critically ill
dogs. Results of the immunologic assays are pending.
Student Support: Stipend for Michael Schacher is supported by an endowment established by IDEXX-BioResearch
250
Seana M. Schade, Patrick Venta, Vilma Yuzbasiyan-Gurkan
Michigan State University College of Veterinary School (Schade, Venta, Yuzbasiyan-Gurkan)
Certain reptile species, especially turtles and tortoises, sometimes experience severe neurotoxic effects (including lethargy,
paralysis, coma and death) when given therapeutic doses of the anti-parasite drug ivermectin. The cause of this sensitivity
has yet to be determined. Similar symptoms are experienced by certain breeds of herding dogs when given the drug. This
is due to a mutation in the MDR1 gene. This study aims to compare the sequences of the MDR1 gene in several species of
ivermectin sensitive animals to those that are not as sensitive to determine if it is the causal gene in reptiles. To do this, the
predicted amino acid sequences for the MDR1 gene in all sensitive species for which sequences are available (the painted
turtle, Chinese softshell turtle, green sea turtle, and alligator) were aligned with those of all reptile species thought to be
insensitive, (the anole, garter snake, and Burmese python). The encoded protein sequences were aligned with the nearest
outgroup species for which the MDR1 gene had been sequenced (the chicken), as well as the human MDR1 amino acid
sequence. From this alignment, several amino acids and amino acid deletions were found to be unique to the ivermectin-sensitive species compared to the unaffected species. Many of these unique amino acid residues are located in structurally
significant regions of the protein. However, the associations by themselves do not prove causation. Further studies are underway to determine the significance of these changes and how they may contribute to the ivermectin intolerance observed
in these species.
Research Grant: Michigan State University College of Veterinary Medicine funds
Student Support: Morris Animal Foundation/Michigan State University Graduate School Fellowship
Gretchen Scheffe, Jessica Romanet, and Jeffrey A. Yoder
North Carolina State University College of Veterinary Medicine, Department of Molecular Biomedical Sciences
and Comparative Medicine Institute, NCSU CVM, 1060 William Moore Drive, Raleigh, NC
Cytokines are secreted proteins that are vital for cellular communication during immune responses. Cytokines trigger the
activation of immune cells including the release of free radicals, nitric oxide, and antimicrobial contents. Cytokine activation is essential to host survival, yet when unchecked, can result in a wide range of illnesses including septic shock, cancer
growth and autoimmune diseases. Thus, understanding the factors that mediate cytokine release is not only critical for
understanding the cellular biology of the immune response, but also is crucial for determining new therapeutic targets for
suppressing cytokine production. The TMEM150a gene encodes a membrane protein of unknown function that is highly
conserved across vertebrate species. Our lab has determined that loss of TMEM150a results in increased cytokine release
in a human cell culture system. We hypothesize that TMEM150A is a negative regulator of cytokine production and release
and that this function is conserved across all vertebrate species. In order to test this hypothesis, TMEM150a was knocked
down in zebrafish embryos with morpholino antisense oligonucleotides. Larvae were then exposed to an immune agonist
(Pam3CSK4) and RNA was isolated at 96 hours post fertilization/24 hours post immune stimulation. RNA was reverse
transcribed into cDNA and cytokine transcript levels will be measured by quantitative PCR. The demonstration that loss of
TMEM150a results in increased cytokine expression in both human cell culture and in zebrafish larvae would validate our
hypothesis and justify future experiments to elucidate the mechanism by which TMEM150a regulates cytokine production.
Research Grant: North Carolina State College of Veterinary Medicine
Student Support: IBRTP/T35 Scholars Program NIH Grant #T35OD011070
251
Alex P. Schenk, Cheryl A. Beatty, Ann M. McGrath, Sheilah A. Robertson, and Andras M. Komaromy
Michigan State University CVM, East Lansing, MI (Schenk, Robertson, Komaromy). Capital Area Humane Society,
Lansing, MI (Beatty, McGrath).
Anecdotal data obtained from shelter veterinarians tells of a phenomenon in which cats undergoing sterilization surgery
develop hyphema during recovery. Hyphema, defined as blood in the anterior chamber of the eye, can cause secondary
glaucoma and decreased visual capacity potentially leading to blindness. Although hyphema can be triggered by numerous
other factors such as trauma or low intraocular pressure (IOP), there are no previous formal reports of spontaneous hyphema
being linked to sterilization surgery. The aim of this study was to confirm reports of spontaneous hyphema in cats and to lay
the groundwork for future investigations about underlying mechanisms. In Part 1, short surveys were sent to veterinarians
in shelters as well as High-Quality High-Volume Spay-Neuter (HQHVSN) practices, and 20 responses were collected. Of
the 20 survey responses, 9 reported having witnessed post-sterilization hyphema at some point during their career. In Part 2,
cats to be spayed/neutered at the Capital Area Humane Society (CAHS) underwent ophthalmic exams, including tonometry,
before and after surgery. One cat out of 191 examined (~0.5%) developed hyphema following surgery during the 8-week
duration of this study. Survey data and first-hand experience support the existence of spontaneous hyphema following sterilization surgery in cats.
No evidence for sexual dimorphism of nitric-oxide (NO) mediated vascular control in rat skeletal muscle
Michael J. Schettler, Jesse C. Craig, Trenton D. Colburn, K. Sue Hageman, David C. Poole, Timothy I. Musch
Kansas State University CVM, Department of Anatomy and Physiology, Manhattan, KS
Gender disparity for cardiovascular disease (CVD) exists with premenopausal females evidencing a reduced risk for CVD
compared with age-matched males. Female estrogen levels, which decrease at menopause, may improve vascular function
and protect against CVD. The mechanistic basis for such estrogen-conferred protection is thought to be facilitated by NO
synthase (NOS) and enhanced NO (a major endothelial-derived vasodilator). We hypothesized that, if estrogen increases
NO bioavailability, female skeletal muscle would exhibit a fundamentally different balance between O2 delivery and utilization; especially during muscle contraction. To test this hypothesis, the spinotrapezius muscle of rats (Male=5; Female=5)
was isolated surgically and electrically stimulated at 1 Hz. Oxyphor G4 was injected into the muscle and phosphorescence
quenching employed to determine the temporal profile of muscle interstitial space O2 partial pressure (PO2, determined
by O2-delivery/utilization ratio). This was performed under 3 conditions: control, 300mM sodium nitroprusside (SNP)
superfusion, 1.5mM L-arginine methyl ester (LNAME; NOS blockade) superfusion. No male-female PO2 differences
were evident under any experimental conditions at baseline (CON: 19.561.5 vs. 16.762.1; SNP: 34.164.8 vs. 36.163.8;
LNAME: 13.862.0 vs. 15.561.9 mmHg; all p>0.05) or in DPO2 during contractions (CON: 10.161.4 vs. 10.361.7; SNP:
15.461.2 vs. 15.762.4; LNAME: 8.061.6 vs. 9.861.5 mmHg; all p>0.05). These results refute the presence of enhanced
estrogen-mediated NO-signaling in females. Therefore, at rest and during muscle contractions, the effect of estrogen on NO
bioavailability and vascular control is insignificant or redundant to other vasodilatory pathways.
Research Grant: National Institutes of Health HL-2-108328
252
Feasibility of viable synovial engineered transplants for paracrine influence on cartilage regeneration
Logan M. Scheuermann, Erin F. Pinnell, Nathalie A. Reisbig, Hayam A. Hussein, Alicia L. Bertone
Synovium, a tissue lining the inside of joints, synthesizes growth factors that are anabolic to articular cartilage and improve shock-absorbing matrix. Our objective was to recapitulate intrinsic signaling with viable engineered synovial grafts
designed to overexpress the growth factor BMP-2 and transplant the grafts adjacent to injured cartilage. Frozen 293A cells
were thawed, expanded and used to propagate adenoviral (Ad) BMP-2 vector. Cryopreserved, viable equine synoviocytes
were thawed, cultured and high numbers were transduced with AdBMP-2 to seed on prepared decellularized synovial scaffolds. ELISA confirmed the living grafts for BMP-2 overproduction. Viable cells (1x106), measured by flow cytometry, were
driven by serum gradient to engraft in the scaffolds. Living grafts were secured to synovium adjacent to cartilage in cadaver
rat knees. We successfully produced a >90% viable synovial graft with a mean of 1x106 cells that overproduced and released
soluble BMP-2 (mean 1.04x10-5 ng/cell). Grafts were transplanted subpatellar using a standard surgical approach that can
be transposed to an arthroscopic approach and lie directly appositional to cartilage for potential paracrine influence on cartilage regeneration. Our work is the first to report a method for novel synovial engineered tissue grafts to potentially promote
cartilage regeneration, as opposed to extensive literature on the, as of yet, poorly successful cartilage grafts for cartilage
replacement. We documented the feasibility of this strategy for knee application that may benefit humans and animals with
knee cartilage damage and osteoarthritis. Our future goal is to move to a clinical model of knee injury in dogs and horses to
validate cartilage influence.
Research Grant: College of Veterinary Medicine Research Council Grant
Effects of calorie restriction on radiation resistance of reserve intestinal stem cells
Jenna Schoenberger, Maryam Yousefi, and Christopher Lengner
Calorie restriction (CR) promotes regeneration of the intestinal epithelium in response to injury such as radiation enteropathy associated with cancer radiation therapy, Celiac disease, ischemia-reperfusion injury, and autoimmune enteropathies for
example. Acute injury leads to atrophy of the intestinal villi and subsequently, malabsorption and potentially dehydration
and cachexia. In response to injury, epithelial regeneration is mediated by activation of a rare, injury-resistant quiescent
reserve intestinal stem cell (ISC). This study will provide important insight into how nutrient intake modulates the ability
of these reserve ISCs to respond to injury and reestablish function of the villi. We have demonstrated that activation of the
mTORC1 nutrient sensing-complex is sufficient to drive ISC activation, and that premature activation and cell cycle entry
of reserve ISCs sensitizes animals to epithelial injury. Conversely, CR is known to have protective effects on the intestinal
epithelium in response to similar injuries. Thus, we hypothesize that CR preserves ISCs in their quiescent state, rendering
them injury resistant, therefore, enhancing the ability of the intestinal epithelium to respond to acute injury. We will test
this hypothesis by 1) assessing the state of ISC quiescence (residence in the G0 state outside of the cell cycle) in response to
CR, 2) assessing mTORC1 activation in reserve ISCs of calorie-restricted mice, and 3) assessing the effects of CR on DNA
damage susceptibility in reserve ISCs. If our hypothesis is correct, the results will have immediate clinical ramifications by
ameliorating intestinal injury, particularly acute radiation enteropathy, through dietary intervention prior to radiotherapy.
Research Grant: R01 CA16865 and P30DK050306
Student Support: National Institute of Health Merial Veterinary Scholars Program
253
Molecular etiology of intrahepatic portosystemic shunts in the Nova Scotia Duck Tolling Retriever
Kelly Schrock, Emily Brown, and Danika Bannasch
Department of Population Health and Reproduction, School of Veterinary Medicine, University of California Davis,
Davis, CA
Intrahepatic portosystemic shunts are a congenital condition characterized by inappropriate vascular development of the
portal vein resulting in blood from the intestines bypassing the liver and entering directly into systemic circulation. Consequently, dogs with this condition present with hepatic encephalopathy due to a build-up of toxic metabolites in systemic
circulation. Preliminary research in the Bannasch laboratory identified a 1.9 Mb region of association on chromosome 18
in the Nova Scotia Duck Tolling Retriever (NSDTR), which included a compelling candidate gene, VEGF-B. Subsequent
whole genome sequencing using 2 NSDTR intrahepatic portosystemic shunt cases and 99 unaffected controls yielded 119
segregating SNPs and 39 segregating small insertion/deletions in the region, however no segregating variants were present
within VEGF-B. Therefore, these variants will be investigated for continued segregation in a larger cohort of 200 unaffected
dogs. Additionally, 7 segregating large insertion/deletions in the interval were identified in the 2 sequenced cases. These
will be genotyped using PCR to investigate segregation in a larger cohort of NSDTR cases and controls. The purpose of
this study is to narrow down the number of segregating variants previously found using whole genome sequencing by genotyping additional cases and controls in the NSDTR breed in order to pinpoint the causative mutation on chromosome 18
responsible for intrahepatic portosystemic shunts.
Research Grant: Bannasch CCAH Portosystemic Shunt I 36194 and Bannasch CCAH Portosystemic Shunt II 42983
Student Support: STAR Program at UC Davis School of Veterinary Medicine, Merial Veterinary Scholars Program
Paraventricular nucleus neurons target nucleus tractus solitarii via CRH receptors and oxytocin in hypoxia
Rachael N. Schulte, Brian C. Ruyle, and Eileen M. Hasser
Department of Biomedical Sciences, College of Veterinary Medicine, University of Missouri, Columbia, MO
The arterial chemoreflex functions to control blood O2 levels during hypoxia (Hx). Chemoafferents project to the nucleus
tractus solitarii (nTS); the nTS integrates this information and sends efferents to the ventrolateral medulla (VLM) and the
paraventricular nucleus (PVN), which are critical for full cardiorespiratory responses to Hx. The efferent pathway from the
PVN that mediates chemoreflex effects is not fully understood. Hypoxia activates (Fos immunoreactive, IR) PVN neurons
projecting to the nTS. PVN-nTS projections express primarily oxytocin or CRH; most projections activated by Hx express CRH. Thus the PVN could contribute to cardiorespiratory chemoreflex responses by activating CRH receptors (likely
CRFR2) in the nTS. CRFR2 co-localizes extensively with oxytocin fibers, and also with synaptophysin, indicating CRFR2
may act by presynaptically modifying oxytocin release. We hypothesized that CRFR2- and oxytocin-expressing puncta
make close appositions onto nTS neurons that are activated in Hx, including VLM-projecting neurons. To test this, fluorescence immunohistochemistry (IHC) targeted CRFR2, HUC/D (neuronal marker), and Fos, or oxytocin and Fos, in brainstem
sections from rats subjected to Hx (10% O2) or normoxia (21% O2). VLM projecting neurons were identified by the retrograde tracer CTB injected into the VLM. Preliminary data suggest that CRFR2-expressing fibers make close appositions
to nTS neurons activated by Hx (indicated by CRFR2-, HUC/D-, and Fos-IR). However, few activated VLM-projecting
neurons exhibited oxytocin-IR puncta. These findings suggest CRH projections from the PVN may influence chemoreflex
function by activating non-RVLM-projecting nTS neurons via CRFR2 and oxytocin.
Research Grant: NIH HL98602
254
Down stream effects of bovine respiratory disease complex on myocardial gene expression
Laura E. Schulze, David Threadgill, Ty Lawrence, Rocky Ward
West Texas A&M University,(Schulze) Texas A&M University, Veterinary Pathobiology, College Station, Texas
(Threadgill); Department of Agricultural Sciences and Beef Carcass Research Center, West Texas A&M University
(Lawrence); Conservation Genetics Laboratory, West Texas A&M University, Canyon Texas (Ward)
Bovine respiratory disease complex (BRDC) is an infectious disease of cattle that causes a $1 billion economic loss yearly
due to death, reduced performance, and costs of vaccinations and treatment modalities. Biological consequences of BRDC
include reductions in feed efficiency and rate of gain in the feedlot, as well as reproductive performance, milk production,
and longevity in the breeding herd. Although no clear link has been established with cattle, viral infections have been associated with cardiomyopathy in humans. Our focus is to determine if there are molecular changes in the heart that contribute
to, or protect from, cardiomyopathy in response to BRDC. This study used 60 cardiac apices flash frozen from an abattoir in
the Texas panhandle. Lungs of all cattle were evaluated during harvest to assess the presence and severity of both pneumonic lesions in the anteroventral lobes and pleural adhesions to rate lung pathology of each animal as healthy (H), moderate
(M), and severe (S). Samples from each apex were processed for histological analysis and RNA was extracted for molecular
analysis. Bar-coded RNA sequencing libraries were prepared and sequenced on a Illumina NextSeq 500. Pooled libraries
were sequenced using 75x75 paired-end targeting 20 million reads per sample. We anticipate cardiac apices will demonstrate differential gene expression in diseased as opposed to the healthy cattle. Further studies will be needed to determine
if differential expression to find candidate genes to predict cattle resistance.
Student Support: Texas A&M University College of Veterinary Medicine & Biomedical Science BRIDGE program
Intraperitoneal ethanol injection for humane euthanasia in zebra finches (Taeniopygia guttata): a pilot study
Chelsea J Schuster, Nathaniel S Kollias, Wendy O Williams, Elizabeth L Buckles, Erin K Daugherity
Faculty of Veterinary Medicine, University of Calgary, Calgary, AB, CAN (Schuster); Cornell Center for Animal
Resources and Education, Cornell University, Ithaca, NY, USA (Kollias, Williams, Daugherity);
Department of Biomedical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, USA (Buckles)
Current euthanasia techniques for avian species are extrapolated from efficacious methods used in mammals, such as CO2
gas euthanasia. However, euthanasia in avian species requires special consideration due to distinct anatomical differences
from mammals. A recent study indicated that intraperitoneal (IP) injection of 70% ethanol was an efficacious, efficient,
cheap, and humane alternative to conventional euthanasia techniques in mice. The objective of this study was to determine
whether 0.5mL of 70% of ethanol is efficacious and humane for IP euthanasia in zebra finches. Twelve cull zebra finches
were obtained at Cornell University, and were block randomized by sex to receive an IP injection of either 0.5mL of 0.9%
sodium chloride, or 0.5mL of 70% ethanol. Finches were placed into a clear behavior box and were video recorded for
5 minutes pre- and post-injection. Blinded and randomized retrospective analysis of multiple behavioral parameters was
completed for all videos. Blood glucose measurements, gross necropsy, and histological analysis were performed following
euthanasia.
Research Grant: Grants for Laboratory Animal Science
Student Support: American Society of Laboratory Animal Practitioners and Pfizer Inc.
255
Effects of deworming and vaccination on seroresponse to BVDV1 and BHV1 in stocker calves
Jenna A. Scott, Brandi Karisch, Amelia R. Woolums, John Blanton, Ray M. Kaplan, William Epperson,
and David R. Smith
College of Veterinary Medicine, North Carolina State University, Raleigh, NC(Scott) Department of Animal and Dairy
Science (Karisch, Blanton) and College of Veterinary Medicine (Woolums, Epperson, Smith), Mississippi State
University, Mississippi State, MS College of Veterinary Medicine, University of Georgia, Athens, GA(Kaplan)
Recently weaned calves moving through market channels into growing or feeding systems may not be immunocompetent at
arrival. The objective of this study was to evaluate the effects of deworming, time of vaccination, and health characteristics
on arrival on seroresponse to BHV1 and BVDV1. Calves (n=80) were blocked by d-3 fecal egg count (FECd-3) and weight
(BWd-3) and assigned to 20 pens (4 calves/pen). Pens were randomly assigned to treatments in a 2 x 2 factorial design of
deworming (oral dewormer at d0 (DWM) or not (NoDWM)) and vaccination (5-way MLV respiratory and 7-way clostridial
vaccines at d0 and 56 (VAC) vs. d56 only (VAC56)). Serum was collected at d0, 28, 56 and 85; log titers and change in log
titers from d0-28, 28-56 and 56-85 were analyzed using linear regression. Main effects were vaccination and deworming
with random effect of pen and covariates of sex, FECd-3, BWd-3 and fever at d0. Calves with higher FECd-3 values had
lower BVDV1 titers at d0 (P = 0.05). From d0 to 28 VAC calves had, on average, a 5.360.8 greater rise in BVDV1 titers
compared to VAC56 calves (P < 0.0001). From d28 to 56 VAC cattle had a 0.960.4 greater increase in BVDV1 titers than
VAC56 cattle (P < 0.0322). From d56 to 85 VAC56 calves had a 4.460.8 greater increase in BVDV1 titers compared to
VAC calves (P < 0.0001). There were no factors which explained arrival BHV1 titers. From d0 to 28 VAC calves had a
2.260.5 greater change in BHV1 titers than VAC56 calves (P < 0.0001). BHV1 titer change from d28 to 56 and d56 to 85
did not differ by any factor. These results indicate that stocker cattle do respond to vaccines given at arrival. Seroresponse
to vaccines differ by pathogen, and deworming does not appear to affect seroresponse.
Research Grant: Mikell and Mary Cheek Hall Davis Endowment
Student Support: Merial and Mississippi State University College of Veterinary Medicine
Identification of Arginine Vasopressin producing neurons in the amygdala of domestic cats
Amanda L Seelman, Farshid M Shahriar, Wael Khamas, Gagandeep Kaur
Western University of Health Sciences, College of Veterinary Medicine, Pomona, CA
Behavior problems including inappropriate elimination are one of the top reasons for relinquishment of domestic pet cats.
Research has shown that the amygdala is one of the most significant brain structures involved in behavior control and modulation. The presence of arginine vasopressin (AVP) producing parvocellular neurons has been documented in the amygdala
of various species, indicating its role in behavior. AVP has been shown to modulate aggression, flank marking, anxiety like
and other behaviors in some rodent species. However, the presence and role of AVP in the amygdala of domestic cats has
not been studied. In the present study, we successfully identified the presence of AVP producing neurons in the amygdala
of domestic cats. Immunohistochemistry was performed on 30 mM freely floating brain amygdala sections from six cats
euthanized for reasons unrelated to this study. Anti-AVP antibody from Phoenix Pharmaceuticals (1:10,000 dilution) was
used. AVP-producing parvocellular neurons were identified in the amygdala of all six cats. These results demonstrate the
presence of this known behavior-modifying neuropeptide in the feline amygdala. Future work will focus on quantifying
AVP-producing neurons in the feline amygdala and eliciting any sexual dimorphism present.
Research Grant: Intramural WesternU
Student Support: WesternU Summer Research Fellowship, from the office of the VP of Research and Biotechnology
256
Two trials to investigate techniques for canine Vitamin D supplementation
Tyler Senft, Robert Backus, Lauren Young
Department of Veterinary Medicine and Surgery, College of Veterinary Medicine, University of Missouri,
Columbia, Missouri
Vitamin D supplementation is being studied as a potential cancer preventative. Some trials have indicated Vitamin D2 in oil
solution may not be the most effective method to raise serum 25(OH)D levels, the indicator of Vitamin D status. Anecdotal
reports have indicated that canines may have low bioavailability to Vitamin D when given in oil capsules, and that 25(OH)
D is significantly more potent than Vitamin D in raising 25(OH)D levels. The purpose of these studies was to develop an
improved method for increasing serum 25(OH)D levels by comparing the bioavailability of Vitamin D2 given in oil versus
ethanol solutions on food, and by testing the stability of ethanol-dissolved 25(OH)D3 on a semi-moist dog treat. Two Beagle-Chinese Crested cross-bred adult dogs were supplemented with a daily 2.3 mg/kg of BW0.75 dose of Vitamin D2 in different vehicles, olive oil and ethanol. Serum 25(OH)D2 levels will be measured once per week using reverse phase HPLC.
I expect that both Vitamin D2 given in ethanol and in oil will raise serum 25(OH)D2 levels at a similar rate. To test the
stability of 25(OH)D3, dog treats spiked with 2 mg 25(OH)D3 were placed in either a refrigerator or at room temperature.
25(OH)D3 levels were measured on the treats once per week using normal and reverse phase HPLC. I expect that 25(OH)
D3 concentration on both room temperature and refrigerated treats will remain consistent over the period tested. If the results of both studies are as expected, it will suggest that supplementing dogs with 25(OH)D3 on a semi-moist treat may be
an effective mechanism to raise serum 25(OH)D3 levels, and that either giving Vitamin D2 in an ethanol solution or oil on
food may be equally effective at raising serum 25(OH)D levels.
Adipose AKT2 controls circulating free fatty acids and systemic insulin resistance
Abigail L. Shearin, Paul M. Titchenell, Qingwei Chu, Patrick Seale, Morris J. Birnbaum
Institute of Diabetes, Obesity, and Metabolism, Perelman School of Medicine, University of Pennsylvania,
Philadelphia, PA
The tightly controlled physiological processes within insulin-responsive tissues are disrupted in insulin resistance (IR)
and Type 2 Diabetes (T2D). Adipose tissue is a critical regulator of energy homeostasis and adipocyte metabolism is perturbed during insulin resistance. Evidence suggests that disrupting insulin signaling in adipocytes leads to systemic insulin
resistance. The proposed mechanism linking adipose signaling and insulin resistance is a decrease in insulin-stimulated
glucose uptake. Studies suggest that a decrease in adipocyte glucose uptake results in hepatic insulin resistance. The adipose glucose transporter Glut4 is largely intracellular until the serine/threonine kinase AKT inactivates the Rab GTPase
that maintains GLUT4 at the lysosome, allowing GLUT4 to translocate to the cell membrane. AKT is a central regulator of
insulin signaling, though its function in vivo specifically in adipose tissue remains largely undefined. In this study, we used
the AdipoQ-Cre to delete Akt2 (Adipo-AKT2 KO) selectively in the adipocyte. Adipo-AKT2 KO mice exhibit adipose and
systemic insulin resistance. We asked if the observed defect in insulin-stimulated glucose uptake is sufficient to cause the
hepatic insulin resistance by comparing our model to mice with AdipoQ-Cre driven deletion of Glut4. Under our conditions,
we found that loss of glucose uptake alone in the adipocyte is not sufficient to cause hepatic insulin resistance. We show
that a defect in insulin’s ability to fully suppress lipolysis causes an increase in circulating FFAs, providing further support
for the model that elevated circulating FFAs lead to hepatic insulin resistance, even in the presence of intact hepatic insulin
signaling.
Research Grant: NIDDK F30 DK100123
Student Support: VMSTP
257
Catherine Si, Rakhi Gupta, Vishnu C. Ramani, Stefanie S. Jeffrey
College of Veterinary Medicine, North Carolina State University, Raleigh, NC (Si) and Department of Surgical Oncology,
Stanford University, Stanford, CA (Gupta, Ramani, Jeffrey)
Patient-derived orthotopic xenografts (PDOX) involve the implantation of human tumors into analogous organs of mice.
Studying these models can uncover remedies of cancers and reduce human testing. But what if a PDOX has transformed histologically and in biomarker expression? Does this model accurately reflect the cancer initially implanted? We hypothesize
passaging a PDOX of breast cancer SUTI097 over several generations of mice has selected for an aggressive subset of tumor
cells from a once heterogeneous population. Further, we hypothesize this subset represents a rare metaplastic breast cancer
(MBC) having no effective treatment. We investigate a gene marker and expression levels to define whether this xenograft
retains previous characteristics and exhibits features of MBC. Results from PCR amplification of PIK3CA gene mutation
and RT-PCR for expression of MBC genes are pending. We also want to analyze SUTI097 and other models at the level of a
single cell. To produce a pure suspension of viable single cells, we perform fine needle aspirates (FNA) of xenograft tumors
ex vivo and investigate the efficacy of various enzymatic digestions as well as a physical strainer. Ten minute enzymatic incubations and the use of accutase seem most effective in separating cells and maintaining viability. However, both straining
and enzymatic digestion do not fully remove debris and clusters of cells. From this study, we hope SUTI097 to be a valuable
model of MBC and emphasize the importance of continued vigilance for markers of disease in PDOX models. We want to
evaluate PDOX on a single cell basis in order to characterize mixed tumor cell populations and their drug susceptibilities
with the goal of combating MBC and other cancers.
Research Grant: NCI IMAT Grant R21CA177447 [or] Komen Foundation SAB1500003 [or] Andrew and Debra Rachleff
Cancer Research Fund [or] John and Marva Warnock Research Fund
Schistosomes recruit host regulatory proteins to resist complement attack
Giles Siddons, Akram Da’darah, Patrick Skelly
Department of Infectious Disease and Global Health, Cummings School of Veterinary Medicine at Tufts University,
North Grafton, MA
Schistosomes are parasitic worms capable of surviving in their hosts’ blood for years without being destroyed by the immune system. The mechanisms they employ to do this remain unclear. Here we show that although recognized and attacked
by the innate immune system, schistosomes foil this attack in part by degrading a key protein in the complement system,
C3. Futhermore, this degradation appears to be mediated by the host regulatory protein Factor I.
Research Grant: NIH (5T35OD010963-07)
Student Support: Unknown
258
Melissa Siegrist, Marcia Hart, Craig Franklin, Aaron Ericsson
MU VRSP (Siegrist); Comparative Medicine Program, Dept of Veterinary Pathobiology, University of Missouri CVM,
Columbia, MO (Hart); MU Metagenomics Center, MU Mutant Mouse Resource and Research Center, Dept of Veterinary
Pathobiology, University of Missouri CVM, Columbia, MO (Franklin, Ericsson)
Differences in gut microbiota (GM) have been shown to modulate many mouse models of disease including models of
colorectal cancer, inflammatory bowel disease, and neurological disorders. During development, the GM of most species
increases in stability and diversity as the animal ages. With adulthood, GM stabilizes until advanced age when it again
becomes unstable and loses diversity. Little is known about early life mouse GM and how early differences in composition
and diversity impact severity and development of disease models. The focus of this study is to monitor the development of
GM profiles of varying composition and diversity. To this end, we are evaluating cecal, colonic and fecal GM in 1, 2, 3, and
6 week old CD1 pups born to mothers with differing vendor-specific GM profiles referred to as GMJAX (originated from
the Jackson Laboratory), GMTAC (originated from Taconic) and GMHSD (originated from Harlan/Envigo). We anticipate
that mice will first be populated with maternal bacterial species of the phylum Firmicutes and that as they age, diversity
will increase with the vast majority of species obtained from their respective dams. We also expect that GM diversity will
reflect maternal GM diversity with pups obtaining GMJAX and GMTAC having lowered diversity throughout development
as compared to pups obtaining GMHSD. Findings from this study will help correlate maturation of GM with differences in
development and severity of a number of diseases that occur later in life. Such correlations will lead to studies that assess
the efficacy of early life GM modulation on preventing disease.
Research Grant: National Institutes of Health (NIH) U42OD010918-18
Student Support: an endowment established by IDEXX-BioResearch
Is there evidence for canalithiasis or cupulolithiasis as an etiology of canine idiopathic vestibular disease?
Anna L. Sirochman and Elizabeth W. Howerth
DVM Candidate, Class of 2018 (Sirochman), College of Veterinary Medicine, University of Georgia, Athens, GA;
Department of Pathology (Howerth), College of Veterinary Medicine, University of Georgia, Athens, GA.
Benign paroxysmal positional vertigo (BPPV) is an inner ear disorder in people characterized by nystagmus, nausea, episodes of vertigo, and dizziness. At present, the accepted etiology involves otolith detachment and migration into the semicircular canals (canalithiasis) or deposition onto the cupula (cupulolithiasis), causing disorientation in the patient. Physicians
utilize head positioning techniques, namely Epley and Semont maneuvers, to dislodge the deposits. Veterinarians occasionally diagnose idiopathic peripheral vestibular disease (IPVD) in geriatric dogs characterized by nystagmus, loss of balance,
head tilt, and circling. Therapy is currently limited to symptomatic management until clinical signs resolve. In 2014, it was
reported that the maneuvers used in treating human BPPV were adapted and employed in canine patients with IPVD and
that expedited improvements in clinical signs were seen. This proof of concept study aimed to develop a method for the
histopathologic examination of the canine inner ear for evidence of canalithiasis or cupulolithiasis. To develop our technique we collected the petrous temporal bones from 8 young beagles and 18 geriatric dogs from the necropsy facility of the
Athens Veterinary Diagnostic Lab at the University of Georgia, College of Veterinary Medicine. Samples were decalcified
using several reagents and processed using both JB-4 resin and paraffin embedding. We determined that the use of formic
acid decalcification solution, paraffin embedding, and sections taken at 10mm yielded the most reliable results. Using this
technique, we plan to examine samples taken from 30 geriatric dogs in order to determine whether or not there is evidence
of canalithiasis or cupulolithiasis in dogs.
259
Examination of genetic variation in CD18 of cattle as a marker for resistance to respiratory disease
Alexis Sirois, Udaya DeSilva, Marie Montelongo, John C. Ritchey, Jerry W. Ritchey
Oklahoma State University CVHS, Stillwater, OK
Bovine respiratory disease (BRD) is an important cause of morbidity and mortality in feedlot cattle. Although the pathogenesis of BRD is complex, Mannheimia haemolytica plays an important role. An important virulence factor for M. haemolytica is leukotoxin (Lkt), which binds and signals through the bovine beta-2-integrin CD18 on immune cells resulting
in dose dependent effects ranging from activation and release of pro-inflammatory cytokines to cell lysis. Although Lkt can
bind, it does not induce signal or effects through non-ruminant CD18 ostensibly because of differences in integrin structure
related to interspecies genetic variation. Therefore, a hypothesis was proposed that individual genetic variation within cattle
expressed as differences in the CD18 gene may result in less or more efficient binding to Lkt and thus relate clinically to
resistance or susceptibility to BRD. To test this hypothesis, RNA was extracted from 137 blood samples collected from feedlot cattle in Oklahoma, Kansas and Texas. The cattle were clinically derived from two groups: Group A, those animals that
remained healthy through the feeding period and, Group B, cattle that exhibited respiratory disease. A segment of the CD18
gene previously shown to be critical for Lkt signaling (corresponds to amino acids 541-581) was amplified by RT-PCR, the
product isolated, purified and sequenced. Sequence results are currently pending and results will be presented.
Research Grant: This work used the Immunopathology Core Program supported by the National Institute of General Medical Sciences under Award Number P20GM103648.
Frequency and complexity of arrhythmias in rhesus macaques with and without hypertrophic cardiomyopathy
Taylor Slabaugh, Yu Ueda, Eric Ontiveros, Christina Cruzen, Jeffery Roberts and Joshua Stern
1. Department of Medicine & Epidemiology, University of California Davis, Davis CA, USA (Slabaugh, Ueda, Ontiveros,
Roberts, Stern) 2. California National Primate Research Center, Davis CA, USA (Cruzen, Roberts, Stern)
Hypertrophic cardiomyopathy (HCM) is a frequently inherited myocardial disease characterized by left ventricular hypertrophy (LVH) and diastolic dysfunction. Mouse models of this disease are frequently studied but are limited in their physiologic relevance. Recently, a naturally occurring, familial, non-human primate model of HCM was described in in rhesus
macaques at the California National Primate Research Center. The high incidence of sudden death (SD) in rhesus macaques
with HCM and no postmortem evidence of thromboembolism or massive myocardial infarction, make an arrhythmic pathogenesis a plausible explanation for these deaths. Based on this information, we hypothesized that rhesus macaques with
subclinical HCM have more frequent and more complex cardiac arrhythmias than age and sex-matched controls. Rhesus
macaques were chosen from a database at the CNPRC and their HCM phenotype was confirmed by echocardiography. Control animals were chosen from the database to match age and gender of the affected group and were required to have a normal echocardiogram. Ambulatory electrocardiography monitors were placed to assess the prevalence of arrhythmias over
24 hours. Upon completion of monitoring the animals were again sedated and the devices were removed. Patient data was
coded to provide a blinded analysis, which is currently underway. Ultimately the frequency and complexity of arrhythmias
as well as assessments of heart rate variability will be performed and tested for statistical differences between the groups.
The data obtained will provide an understanding of the electrophysiological status of rhesus macaques with HCM, and may
provide translational value to expand our understanding of human SD in HCM patients.
Research Grant: Supported by UC Davis Center for Companion Animal Health
260
The role of lidocaine and meloxicam in pain management for bull castrations
Katie Slimack, Dusty Nagy, Pamela Adkins, and Brian Vander Ley
MU VRSP (Slimack), University of Missouri College of Veterinary Medicine, Columbia, MO
Castration in bull calves has been shown to lower average daily gain, which ultimately results in decreased overall production and associated loss of revenue for feedlots. Pain management, including local anesthetics and nonsteroidal anti-inflammatory drugs can improve performance, decrease morbidity and minimize pain related behaviors. We hypothesized that
using a combination of lidocaine and meloxicam will result in optimal pain management as evidenced by increased feed
intake and decreased pain-related movement behavior. To this end, we assessed feed intake and behavior of bulls through
movement following castration and administration of meloxicam, lidocaine, or a combination of both meloxicam and lidocaine. Bulls were chosen at random to receive one of the four treatments, including the control group. To measure behavioral
changes in movement, bulls were tracked with an IceQube© sensor, an accelerometer, that was applied on the rear limb
when castrated. The sensors measured motion, standing, laying down and steps taken, and data was transferred to a computer. Measurements were also taken using the GrowSafe©, a system designed to track feeding behavior. It is expected that
these measurements will show that the castration procedure using both lidocaine and meloxicam together is most beneficial
to the calves and increases feed intake and overall production, while decreasing movement and restlessness. Results of this
study are forthcoming.
Research Grant: Vander Ley Startup Fund
Student Support: College of Veterinary Medicine Agricultural Experiment Station Funds
Genetic investigations of the AMH and AMHR2 genes in canine persistent Mullerian duct syndrome
Michelle M. Smit, Eva Furrow, Katie M. Minor, Peter A.J. Leegwater, Chee K. Lim, Kari J. Ekenstedt
Faculty of Veterinary Medicine, Utrecht University, The Netherlands (Smit, Leegwater) College of Veterinary Medicine,
University of Minnesota, St. Paul, Minnesota, USA (Furrow, Minor, Ekenstedt) College of Veterinary Medicine, Purdue
University, West Lafayette, Indiana, USA (Lim)
Persistent Mullerian duct syndrome (PMDS) is a reproductive disorder in which male dogs develop portions of the female
reproductive tract. Cryptorchidism, and the sequelae of infertility and increased risk for testicular cancer, are important
consequences of PMDS. The anti-Mullerian hormone (AMH) and receptor are involved in the regression of the Mullerian
ducts in male embryos and are therefore essential in mammalian sex determination. The genetic basis for PMDS in Miniature Schnauzers (MS) is a sex-limited autosomal recessive nonsense mutation in the AMH receptor gene, AMHR2. Although
a genetic test has been available since 2009, the prevalence of the mutation in MS is unknown, and its existence in other
breeds has not been reported. Such information is crucial to determining the value of genetic testing in MS and would aid
development of genetic tests for other affected dog breeds. Allele-specific primers were designed to genotype dogs for the
AMHR2 mutation. Genomic DNA of 216 MS (containing one known male PMDS case) and 1 Belgian Malinois (a known
male PMDS case) was tested. The tested MS population had a AMHR2 mutation frequency of 16% and carrier rate of
27%. Besides the 2 known affected male dogs, 2 female dogs were found to be homozygous for the mutation. The Belgian
Malinois case was clear for the MS mutation. Thereafter both the AMH and AMHR2 genes were sequenced in the Belgian
Malinois in an attempt to discover a new mutation. No coding mutations have been identified to date, but sequencing is
still in progress. These findings provide a research-based recommendation to test all MS used for breeding for the AMHR2
mutation. Furthermore, it demonstrates the need for more PMDS research in other dog breeds.
Research Grant: Discretionary funds of co-PI (Furrow)
Student Support: Merial Veterinary Scholars Program and Utrecht University
261
Carolyn Smith, Kevin Keegan, Yoshiharu Yonezawa
University of Missouri College of Veterinary Medicine, Columbia, MO
Laminitis is a significant cause of career-ending lameness in pleasure and performance horses, yet the diagnosis is often
made after irreversible damage to the hoof has taken place. Early detection and treatment help to dramatically improve the
prognosis of a horse diagnosed with laminitis. One subjective clinical sign commonly seen in early onset laminitis is weight
shifting between the forelimbs as the horse attempts to alleviate pain and pressure. Although weight shifting has been well
described in early onset laminitis, the frequency of this behavior between laminitic and non-laminitc horses is unknown.
The normal frequency of weight shifting is also unknown. Knowing the difference in frequency of weight forelimb weight
shifting between laminitic and non-laminitic horses could help detect laminitis earlier, allowing for more rapid intervention and treatment. The objective of this current study is to establish how frequently non-laminitic horses shift their weight
between the forelimb hooves. We hypothesize that normal horses will fall within a range of regular and relatively low frequency weight shifting that will average less than ten times per minute. Healthy (non-laminitic) horses of various breeds,
genders, and ages will wear force shoes held in easy boots on all four hooves for approximately 24 hours while stalled. Data
on the force on each hoof and frequency of weight shifting will be analyzed, and an average range will be established. By
better understanding the weight shifting behavior of sound horses, we hope to characterize the differences in weight shifting
behavior in laminitic horses and help develop a screening tool to detect laminitis before irreversible hoof damage occurs.
Research Grant: The E. Paige Laurie Endowed Program in Equine Lameness supports Dr. Yoshiharu Yonezawa as a
visiting professor.
Student Support: Endowment established by IDEXX-BioResearch
Comparison of molecular methods in the characterization of Malassezia on the skin of healthy and allergic dogs
Courtney Meason-Smith, Sara D. Lawhon, Thierry Olivry, Aline Rodrigues Hoffmann
Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Sciences, Texas A&M
University, College Station, TX (Meason-Smith, Lawhon, Hoffmann). Department of Clinical Sciences, College
of Veterinary Medicine and Comparative Medicine Institute, North Carolina State University, Raleigh, NC (Olivry)
The advancement of DNA sequencing technologies has revolutionized the study of skin commensals, how they contribute to
health and the role they play in disease processes. Recently, next-generation sequencing (NGS) has been applied to estimate
the relative abundance of microorganisms on the skin of animals. The purpose of this study is to use additional methods
including real-time quantitative PCR (qPCR) to quantify Malassezia spp. on skin swabs, and neighbor joining analysis on
NGS data to classify skin-associated Malassezia to the species level. A reference tree was constructed from Malassezia
spp. using the softwares MUSCLE and MEGA. All 38,073 Malassezia ITS-1 sequences were extracted from a previously
published dataset including 106 skin swabs of healthy and allergic dogs. To quantify Malassezia from the skin swabs, a
qPCR using previously published primers specific for Malassezia was performed on the aforementioned dataset, as well as
a dataset comprised of 96 skin swabs from a house dust mite induced canine model of atopic dermatitis. The skin of allergic
dogs without lesions had significantly less Malassezia compared to healthy skin and this was especially true for the axilla.
Dogs exposed to house dust mite showed a reduction in Malassezia from pre-exposure to seven days after lesion development. We expect M. pachydermatis will predominate in skin swabs of dogs, but that other Malassezia species will be present
in smaller quantities. Additional molecular methodologies are proving to be useful in the study of skin commensals, and
further illuminate how these commensals play a role in allergic skin disease.
262
Eosinophil activity is not altered by respiratory syncytial virus infection in the cotton rat
Katelin L. Smith and Stefan Niewiesk
Veterinary Biosciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH
Respiratory syncytial virus (RSV) is the most prevalent cause of infantile bronchiolitis and pneumonia worldwide. Although children are infected by age 2, infection does not induce long-term immunity and reinfection throughout life is
common. Severe RSV infection as an infant significantly increases the likelihood to develop asthma. This connection is
not well understood and has been studied in mice. However, RSV replicates only to low titers in lung tissue of mice. In the
current study we used cotton rats which replicate RSV in the nose and the lung. Neonatal cotton rats, were infected with
RSV and reinfected as young adults to evaluate lung and nasal remodeling. Neonatal lung remodeling was compared to
animals whose first exposure occurred during adulthood. House dust mite antigen (HDM) was used to induce an allergic
response in some animals to mimic natural allergen exposure in children. Allergic responses were measured by the number
of eosinophils in bronchoalveolar lavage and secretion of eosinophil peroxidase. When cotton rats challenged with HDM
were infected with RSV or influenza virus, the number of eosinophils in the lung was not significantly changed compared
to an uninfected group. Influenza virus was used as a control because this viral infection is not known to result in asthmatic
symptoms later in life. The amount of eosinophil peroxidase measured per eosinophil was not significantly different between any of the groups thus we conclude that eosinophil activity is not altered by RSV infection. The effects of neonatal
infection on lung and nasal remodeling is ongoing.
Student Support: NIH T 35 Training Grant
Bacterial killing ability of vampire bats and associations with individual body condition and feeding ecology
Katherine Smith, Daniel J. Becker, Gabor Á. Czirjak, Sonia Altizer
College of Veterinary Medicine, University of Georgia, Athens, GA (Smith); Odum School of Ecology, University of
Georgia, Athens, GA (Becker, Altizer); Leibniz Institute for Zoo and Wildlife Research, Berlin, Germany (Czirjak)
Understanding the immunocompetence of the common vampire bat (Desmodus rotundus), and how it is influenced by
individual and environmental factors, is important for determining the risk of zoonotic infections of bats and potential
transmission to humans and livestock. To achieve this, we optimized a bacterial killing assay to find the proportion of
E. coli killed in vampire bat plasma. Body condition and percent of domestic animals in diet may be indicators of how
frequently bats feed, with more well-fed vampire bats having better immunity. Beta regression models were built to determine associations between resource acquisition and bacterial killing ability (BKA), adjusting for how long plasma was
frozen. A positive association was found between BKA and body condition, stratified by age with adults having a higher
BKA than subadults. In a subset of individuals for which stable isotope data were available, a positive association was
found between BKA and frequency of feeding on domestic animals, with bats in agricultural habitat having higher BKA
than bats in a nature reserve. These findings suggest that well-fed vampire bats have better innate immune defenses, potentially due to lower starvation and ability to invest more energy in immune function. Bats feeding on domestic animals
might also need to invest more in innate immunity due to increased pathogen risk in agricultural sites. Because greater
access to livestock was associated with more feeding on domestic animals, livestock rearing could lower risk of human
bites and cross-species transmission. Future studies should determine if higher BKA in vampire bats is also associated
with decreased infection prevalence of zoonotic bacteria, such as Bartonella or Mycoplasma.
Research Grant: Funding was provided by the National Science Foundation, ARCS Foundation, Sigma Xi, University of
Georgia Biomedical and Health Sciences Institute, Odum School of Ecology, and Explorer’s Club.
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Tricaine methane sulfonate (MS-222) anesthesia in fishes: Relationship to thermal divergence
Kayla Smith, Jeanette Cremer, and Kevin Kleinow
Departments of Veterinary Clinical Sciences (Cremer) and Comparative Biomedical Sciences (Kleinow), School of Veterinary Medicine, Louisiana State University, Baton Rouge, LA
Poikilothermic fish maintain cellular and physiological stability in face of slowly changing environmental temperatures by
the process of acclimation. Changes in membrane composition, organelle density, isozyme shifts and receptor numbers are
central to this process. Acute body temperature change differentially alters the structural and functional consequences of
the compensatory changes enabled by acclimation. This has been known to affect drug pharmacokinetics and pharmacodynamics. Tricaine methane sulfonate (MS -222), a widely accepted fish anesthetic, is often clinically used under temperature
conditions different from acclimation. This project aimed to determine if acute temperature divergence, either above or
below acclimation will differentially impact MS-222 induced stages in the goldfish model. While water temperature colder
than the acclimation temperature accelerated anesthetic progression, warmer water temperature exhibited great variability
and lacked predictability. This study’s findings help to establish a basis regarding clinical temperature strategies, pertaining
to fish anesthesia; a foundation upon which future research may be constructed.
In vitro effects of the chemotherapeutic agent water-soluble paclitaxel on canine mast cell tumor cell lines
Mallory Smith, Keijiro Shiomitsu, Marc Salute, Brandy Winter, Amandine Lejeune
Small Animal Clinical Sciences, College of Veterinary Medicine, University of Florida, Gainesville, FL
Mast cell tumors (MCT) are the most common canine cutaneous malignancy encountered in the dog. Treatment with chemotherapy is recommended if the patient has systemic disease or high grade mast cell tumor(s). While chemotherapeutic drugs
are available to treat MCTs, resistance to these drugs often arises and may lead to disease progression and patient death.
Water-soluble paclitaxel is a newly formulated, taxane-class chemotherapeutic drug conditionally approved by the FDA
for the treatment of mammary carcinoma and squamous cell carcinoma in canines. Due to its broad-spectrum activity and
predictable safety profile, this class of drug is widely used in human oncology. This study evaluated the in vitro activity of
paclitaxel against three canine mast cell tumor (MCT) cell lines after treatment with increasing concentrations of paclitaxel
(0mM, 0.001mM, 0.005mM, 0.01mM, 0.05mM, 0.1mM, 0.5mM, 1mM) at 24, 48, and 72 hours. Mean half maximum inhibitory concentration (IC50) at 72 h for CoMS, LuMC, and TiMC was 0.0335mM, 0.0351mM, and 0.0035mM, respectively.
Caspase-3/7 activity increased in correlation with the IC75, IC50, and IC25 in each cell line which was confirmed by the
terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL; Apo-BRDU kit; BD Biosciences, San Jose, CA)
assay. Paclitaxel also caused G2/M phase arrest in MCT cell lines. Overall, the study found that paclitaxel causes dose and
time dependent MCT cell death through initiation of caspase-mediated apoptosis. This makes paclitaxel a promising chemotherapeutic agent for treatment of canine MCT. Further clinical evaluation would be highly warranted.
Research Grant: University of Florida College of Veterinary Medicine
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VP22 transport from differentiated Neuro2A cells in microfluidic chambers to epithelial cells and vice versa
Richard Smith, Jared Rudd, Katrin Pannhorst, Azharul Islam, Shafiqul Chowdhury
Louisiana State University, Pathobiological Sciences, School of Veterinary Medicine, Baton Rouge, LA
BHV-1 causes respiratory infection, abortion and is associated with bovine respiratory disease complex. Following intranasal infection and initial replication in nasal epithelial cells, the virus enters the sensory nerve endings of the trigeminal
nerve. Thereafter, nucleo-capsid containing tegument proteins, including VP22, encoded by the BHV-1 UL49 gene, are
transported to trigeminal ganglia where the virus establishes lifelong latency. Upon stress, latent virus reactivates resulting
in transportation to nasal epithelium, replication and virus shedding. The alpha herpesvirus VP22 homologs are highly
conserved, and are able to move from cell-to-cell in the absence of other viral proteins. We hypothesized that BHV-1 VP22
can move from the epithelial cells to neurons and vice versa. To test the hypothesis, Dr. Chowdhury’s lab has constructed a
BHV-1 recombinant virus in which VP22 is fused in-frame with a red fluorescent protein (red VP22) as well as a red-VP22
expressing stable MDBK cell line. Neuro 2A cells can be differentiated into neurons in culture when the culture media
are supplemented with retinoic acid and nerve growth factors. In this project my goals have been i) growing the Neuro 2A
cells in the microfluidic chamber system, ii) testing their differentiation into neurons and extension of the axons through
the grooves from the soma to the axonal compartment. If successful, the red-VP22 expressing cell line will be seeded in the
axon compartment to determine whether the red-VP22 protein expressed by the MDBK cells can be transported retrograde
along the axons. In addition, the neurons in the soma chamber will be infected with the red-VP22 expressing virus to determine the anterograde axonal transport.
Student Support: Louisiana State University School of Veterinary Medicine
Sam Smith, Michael Childress, Elaine Baird, Mary Mengel, G. Kenitra Hammac, Christina Wilson
College of Veterinary Medicine; Department of Veterinary Clinical Sciences, Purdue University, W. Lafayette, IN
(Smith, Childress) Animal Disease Diagnostic Laboratory (Baird, Mengel, Hammac, Wilson),
Department of Comparative Pathobiology; Purdue University, W. Lafayette, IN (Hammac, Wilson)
Noviplex duo plasma prep cards are a new micro-sampler product that separates serum from whole blood. They allow
3.5mL of serum to be collected into two small disks that can be used for a variety of testing. These cards only require two
drops of whole blood and hold potential clinical value for blood testing in anemic and smaller patients. Usually, blood tests
require milliliters of blood to perform selective, routine diagnostic tests. A study was conducted to determine the usefulness
of these cards in veterinary clinical practice. Canine heartworm antigen and lactate dehydrogenase (LDH), a potential prognostic biomarker for canine lymphoma, were chosen to assess the efficacy of the Noviplex duo plasma prep cards. Serum
samples collected with Noviplex cards were compared to serum collected by centrifugation of whole blood. Serum extraction from the cards was achieved using phosphate-buffered saline solution, and the activity of LDH and concentration of
heartworm antigen were measured by enzyme-linked immunoabsorbent assays (ELISA). Both tests are colorimetric assays
in which the optical densities were quantitated with a microplate reader. Using Noviplex cards a positive color change can
be observed in both tests but the intensity of the color is usually less than that of a direct serum sample. However the optical
densities have been consistently higher than all negative controls so it is possible for a reference interval to be established.
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A Comparison of Lyme disease Strain Diversity in Michigan Island and Mainland Sites
Lauren Smyth, Jennifer Sidge, Jean Tsao
College of Veterinary Medicine (Smyth), Comparative Medicine and Integrative Biology (Sidge),
Department of Large Animal Clinical Science (Tsao), Michigan State University, East Lansing, MI
Borrelia burgdorferi, the causative agent of Lyme disease, afflicts thousands of people and canines each year. This pathogen
is carried by Ixodes scapularis, the blacklegged tick, which has been spreading in Michigan’s Lower Peninsula. Clinical
manifestations of Lyme disease may be related to genetic variation among different B. burgdorferi strains. We hypothesize
that strains will vary among sites as a function of vertebrate host community composition and invasion history. To test our
hypothesis, we compared the diversity of B. burgdorferi among blacklegged tick populations and animal ear biopsies on
North and South Manitou Islands and several mainland sites. These two islands in Lake Michigan represent two island
ecosystems with differing host compositions from mainland sites, which presumably would form the source of B. burgdorferi strains. Specifically we will compare the diversity of B. burgdorferi among sites targeting a fragment of the 16S-23S
intergenic spacer region. The study revealed that 264 samples were comprised of nine different strains, the most common
of which was HRPW91, composing 33% of all samples. Three strains were only represented once in all of the samples.
Overall, strain diversity by proportion was fairly similar across the different sites. Of note was the strain M11P which was
only found once but was first sequenced from a human biopsy in a persistent infection. Further research should be devoted
towards continued sampling and sequencing of the mainland and South Manitou Island sites as the tick and B. burgdorferi
populations continue to expand and grow in population abundance.
Research Grant: NSF EF-0914476, NIH Grant No T35OD016477 to Michigan State University
Why won’t you eat me? Investigating melanomas’ unique resistance to phagocytosis through RNAi
Kristin M. Snyder, Katie L. Anderson, Jaime Modiano
Department of Veterinary Clinical Sciences, College of Veterinary Medicine, University of Minnesota, St. Paul, MN,
USA; Masonic Cancer Center, University of Minnesota, Minneapolis, MN, USA
Evasion of phagocytosis is a common survival strategy that allows malignant cells to prevent elimination by the immune
system. CD47 blockade to impair “don’t eat me signals” and chemotherapeutic upregulation of known “eat me signals” such
as calreticulin (CRT) and phosphatidylserine (PS) represent potential strategies to enhance the anti-tumor immune response.
The pathways by which melanomas evade phagocytosis are incompletely understood. Previous studies in the lab showed
that neither CD47 blockade nor upregulation of CRT and PS using chemotherapy were able to enhance macrophage-mediated phagocytosis of melanoma cells from humans, mice and dogs. This resistance was not observed in lymphoma cells from
humans, mice or dogs, or in other solid tumors such as canine osteosarcoma, feline mammary adenocarcinoma, mouse Lewis lung carcinoma or human colon carcinoma, suggesting that melanomas possess a unique and uncharacterized resistance to
phagocytosis. Our working hypothesis is that the peculiar resistance to phagocytosis of melanomas is due to the expression
of a heretofore uncharacterized “don’t eat me” signal on the cell surface. We designed a siRNA panel containing 48 genes
whose protein products are expressed on the cell surface of melanoma cells. We will report our results of how knockdown
of these proteins through RNA interference affects macrophage-mediated phagocytosis of melanoma cells.
Research Grant: NIH F30 CA 195973, NIH P30 CA077598, Skippy Frank Fund for Life Science and Translational Research, Donors to the Animal Cancer Care and Research Program of the University of MN
Student Support: Office of Graduate Programs and the College of Veterinary Medicine, University of MN
266
The effect of prenatal alcohol exposure on respiratory regulation during sleep in the early postnatal period
Emily Soltis and Kevin Cummings, PhD
University of Missouri College of Veterinary Medicine, Biomedical Sciences, Columbia, MO
The risk of the Sudden Infant Death Syndrome (SIDS), a leading cause of infant death, is elevated by prenatal alcohol
exposure, but the mechanism for this is unknown. SIDS victims display respiratory instability during sleep, and increased
active sleep (AS; similar to REM sleep), a state characterized by respiratory instability. In addition, SIDS victims have
decreased levels of serotonin (5-hydroxytryptamine; 5-HT), a neurotransmitter involved in the regulation of breathing.
We hypothesized that prenatal alcohol exposure increases SIDS risk by destabilizing breathing in infancy. To address this
hypothesis we studied rat pups at postnatal day 7-8, born from dams that were fed 10% ethanol (EtOH) or pure water as
their sole source of liquid during pregnancy. We used whole-body plethysmography to record the respiratory pattern, and
measured the respiratory period (the time for each breath), tidal volume, as well as the standard deviation of each variable as
indices of respiratory stability. We also determined the time spent in AS using behavior criteria that were confirmed in preliminary experiments with electromyography. Our data demonstrate that exposure to EtOH resulted in increased duration of
AS episodes (EtOH: 100.064.09 sec; control: 80.8561.64 sec), as well as increased the variability of the respiratory period
(EtOH: 0.134 60.016 sec; control: 0.113 6 0.004 sec) and tidal volume (EtOH: 0.04760.003 ; control: 0.03760.002).
These results suggest that prenatal EtOH exposure increases SIDS risk by destabilizing breathing in infancy. Future studies
will address the hypothesis that prenatal EtOH exposure destabilizes breathing by reducing brainstem 5-HT levels.
Research Grant: American Heart Association (Scientist Development Grant to KJC)
Student Support: The Department of Biomedical Sciences, University of Missouri College of Veterinary Medicine
A new incapacitance meter for the measurement of static weight-bearing forces in the hind limbs of mice
Samantha L. Sommer, Michael W. Nolan
NC State College of Veterinary Medicine, North Carolina State University, Raleigh, NC
BACKGROUND: We have developed a new incapacitance meter (IM) that measures hind limb static weight-bearing forces
in mice while allowing them to maintain a natural standing posture. AIMS: The aims of this investigation were three-fold:
1) calibrate the IM and determine the recommended calibration interval; 2) describe hind limb weight distribution for normal mice; and 3) assess the ability of the IM to quantify experimentally-induced hind limb lameness in mice. METHODS:
The device was calibrated using standardized weights and calibration was monitored for 10 days. Normal hind limb weight
distribution data were collected using the IM for 10 days in normal mice. Acute hind limb lameness was induced in a second
cohort of mice via a subcutaneous injection of capsaicin (0.25 mL 0.1% w/v) into the plantar surface of the left hind paw.
Baseline IM measurements were recorded and these measurements were repeated at various time points post-injection. IM
measurements were compared to visual lameness scores recorded at identical time points. RESULTS: Calibration remained
stable for the 10-day period and from 17 to 35 degrees Celsius. On average, 49.9% +/- 1.2% of hind limb force was borne
on the left hind limb in normal mice as measured by the IM, and this distribution did not vary significantly over time. In
mice with capsaicin-induced lameness, IM weight-bearing measurements correlate well with visual lameness scores (r2 =
0.93). CONCLUSION: IM weight-bearing measurements accurately reflected the observed capsaicin-induced lameness.
These results provide proof of concept for the use of this IM to objectively measure static weight-bearing forces in the hind
limbs of laboratory mice.
Research Grant: Didi Cancer Research Fund
Student Support: NC State College of Veterinary Medicine Fund for Discovery
267
Juli Sorenson, Giedre Turner, Craig Franklin, Aaron Ericsson
Veterinary Research Scholars Program, College of Veterinary Medicine, University of Missouri (Sorenson)
Metagenomics Center and Mutant Mouse Resource and Research Center, Department of Veterinary Pathobiology,
College of Veterinary Medicine, University of Missouri (Franklin, Turner, Ericsson)
The importance of the gut microbiota (GM) to human and animal health has undergone close scrutiny within the past decade. Once believed to be of little importance to the overall wellbeing of patients, variations from normal GM composition
and levels have been linked to various illnesses. Mouse models provide an optimal method to study the GM, as mice are
genetically identical and can be maintained in large numbers within a controlled environment. Additionally, the mouse’s
ability to adopt the GM of other species has been well documented, allowing for the mouse to model species that are less
amenable to controlled studies. Although this transfer has been performed from many species, the ability to transfer equine
GM to mice has not been well documented. The purpose of this study is to determine the degree to which equine GM can be
transferred to mice and establish this as a model to study equine disease. 16 FVB/NJ mice were treated with four antibiotics
to deplete their endogenous GM and then gavaged with equine fecal material to create an ‘equinized’ GM. Mouse fecal
samples were obtained at three different intervals: before antibiotic administration (to measure their natural GM levels),
after antibiotic administration (to ascertain depletion of the natural GM), and post-gavaging (to confirm colonization of the
‘equinized’ GM). To determine GM composition, DNA is being extracted and undergoing sequencing and Qime analysis.
We expect that the mice will successfully adopt the majority of the GM seen in the equine, and that this GM will transfer to
subsequent generations creating a sustainable model. Ultimately, our goal is to have ‘equinized’ mice that will yield critical
information on how the equine GM affects disease.
Research Grant: University of Missouri Metagenomics Center
Student Support: The stipend for Juli Sorenson is provided by an endowment established by IDEXX-BioResearch
The effect of more liberal milk allowance in the first week of life on performance of preweaned dairy calves
Janna Sorg, Sandra Godden, Whitney Knauer
Department of Veterinary Population Medicine, University of Minnesota, St. Paul, Minnesota
The health and growth of calves is important to dairy producers as calves represent the future of the dairy operation. One
contributing factor to this is the volume of milk calves receive throughout the pre-weaning period. Benefits such as enhanced immune response, increased growth rate, increased first lactation milk production and improved resistance to diarrheal pathogens have been observed in calves fed increased levels of milk. The objective of this randomized clinical trial is
to investigate the effect of offering a more liberal milk supply from day 1 on calf health and growth, as compared to slowly
increasing the milk allowance over a 1-2 week period. We hypothesize that calves allowed to drink more milk from day 1
will grow faster and be healthier during the pre-weaning period as compared to calves on a traditional ramp up program.
This randomized clinical trial was conducted on 5 MN dairy farms. Newborn calves were weighed at birth, and then randomly assigned to either the control group (standard ramp-up program) or the treated group (liberal milk allowance). Farms
were then visited weekly. Blood samples were collected at week 1 to measure serum total proteins (g/dL). Calf health was
measured through weekly health scores (w1-w3) and producer reported treatment events. Growth was measured through the
collection of hip heights (w1,3,7) and body weight (birth, w3). Calves exited the study at weaning. A total of 463 calves have
been enrolled to date (n=258 treated; n=205 control). Enrolled calves have an average birth weight of 38.8 kg (range: 24.957.2) and an average STP of 5.74 g/dL (range: 4.1-8). Data collection will conclude in late August, with analysis to follow.
Research Grant: Land O’ Lakes
Student Support: Summer Scholars Program
268
Benefits of a therapeutic horseback riding program on veterans suffering from PTSD or traumatic brain injury
Emmi Stabler, Rebecca Johnson, Sandra Crowder, Dorothea Megarani
College of Veterinary Medicine, University of Missouri, Missouri, USA
Frequently, veterans return from combat and suffer from Post Traumatic Stress Disorder and/or a traumatic brain injury.
These diseases can result in a number of symptoms ranging anywhere from anxiety, flashbacks, and emotional numbing,
preventing them from returning to normal life. There is research supporting the idea that THR can help relieve some of
these symptoms; however, little research has been done on veterans. This project examines the efficacy of a human-horse
interaction and systematic therapeutic horseback riding program in reducing these challenges. Anticipated themes include:
psychological wellbeing, healthy interaction and emotional connections. Using a randomized experimental design with repeated measures, veterans chosen for this program were over the age of 18, weighed under 250 pounds, were able to walk
25 feet independently, consented to participate and had approval from their Health Care provider. The Riding Group spent 1
hour, weekly riding the same horse at 1 of 3 accredited riding centers. The activity was directed by a systematic lesson plan.
After each riding session, the veterans were given a riding diary to write down their specific perceptions. Using Colazzi’s
method, positive and negative themes will be identified from these diaries. So far, I believe the veterans perceived the THR
program in a positive way. I anticipate that through further analysis I will extract more specific positive and negative factors.
These factors then can then be used by health promotion advocates to expand similar programs world-wide. Additionally,
this discourse will erect a call for research that draws upon the main theme that companion animals can have a positive
impact on both physical and mental health.
Research Grant: Horses and Humans Research Foundation (Grant #00040568)
Student Support: Supported by the USDA National Institute of Food and Agriculture, Animal Health project 1003471
gd T Cells play a critical role in the inflammatory response to Paramyxovirus infection in mice
Philip B. Stelly, Nagarjuna R. Cheemarla, Antonieta Guerrero-Plata
Department of Pathobiological Sciences (Cheemarla, Guerrero-Plata), College of Veterinary Medicine, Louisiana State
University, Baton Rouge, LA
Human metapneumovirus (hMPV) belongs to the Paramyxoviridae family and is a leading respiratory pathogen causing
severe symptoms such as bronchiolitis and pneumonia in young children, the elderly and immunocompromised patients.
hMPV accounts for nearly 5-10% of hospitalizations in children and remains the most important cause of infant and young
children mortality. Since its discovery in 2001, the regulation of the immune response against hMPV remains largely unknown. We have demonstrated that, specific depletion of neutrophils in vivo using a monoclonal antibody exhibited a significant increase in lung inflammation and a significant increase in the gd-T-cell subset in the lungs compared to neutrophil
competent mice. Moreover, we aimed to identify the contribution of gd-T-cells to lung inflammation induced by hMPV
infection. Our findings demonstrated that gd-T-cells are recruited to the lungs of infected mice during the acute phase of
hMPV infection and gd TCR-/- mice exhibited reduced pulmonary inflammation. In summary, these findings indicate that
gd-T-cells are critical players in the inflammatory responses to this paramyxovirus infection.
Research Grant: NCRR- 5P20RR020159-09, NIGMS- 8P20GM103458-09, Flight Attendant Medical Research Institute
CIA grant
Student Support: NIH T35 T35OD011151-13
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Helminths in mountain gorillas: conservation through intestinal parasite monitoring
Robert J. Stenger, David R. Smith, Gladys Kalema-Zikusoka, and Stephen Rubanga
Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University,
Starkville, MS (Stenger and Smith) Conservation Through Public Health, Entebbe, Uganda (Kalema and Rubanga)
The mountain gorilla (Gorilla beringei beringei) brought to international attention by Dian Fosey’s Gorillas in the Mist is
critically endangered. Through great effort from local peoples, governments, and conservation NGOs the mountain gorilla
population is rising. Yet with rapidly growing local human populations and greater tourist visitation there is an ever increasing risk of zoonotic disease transmission. Using fecal samples collected in a longitudinal monitoring program we looked
for variation in intestinal parasite prevalence, hypothesizing that prevalence would differ by factors which were recorded
at sample collection (gorilla age, gorilla group, month of collection, etc.). Parasite prevalence was determined using fecal
flotation (Anoplocephala, Strongyle, Ancylostoma, and Taenia eggs identified). Our objective was to quantify intestinal
parasite prevalence and test if prevalence varied by the different factors of interest. We looked at the mountain gorilla populations in Bwindi and the Virungas at three different time periods (2005, ‘06, & ‘09). Results of our multivariable logistic
regression indicate that there is statistically significant variation in the prevalence of intestinal parasites within mountain
gorillas (a = 0.10). Parasite prevalence varied by the individual gorilla (Virungas, ‘09) and month of collection (Bwindi and
Virungas, ‘05 & ‘09). The next step is to determine if this variation is due to natural differences in ecology or human induced
factors. These data are currently used to monitor gorilla health. However, a better understanding of the epidemiology of
intestinal parasitism in the mountain gorilla will allow for more informed management decisions.
Research Grant: Supported by the Mikell and Mary Cheek Hall Davis Endowment for Beef Cattle Health and Reproduction
Microbial community structures in E. coli O157 shedding and non-shedding dairy cattle; a preliminary analysis
Chloe M Stenkamp-Strahm, Sheryl L. Magzamen, Zaid O. Abdo, and Craig S. McConnel
Department of Clinical Sciences (Stenkamp-Strahm and McConnel), Environmental and Radiological Health Sciences
(Magzamen) and Department of Microbiology, Immunology and Pathology (Abdo), College of Veterinary Medicine and
Biomedical Sciences, Colorado State University, Fort Collins, CO
Shiga toxin (stx) producing E.coli O157 (O157) infects approximately 100,000 humans annually. Dairy cattle harbor and
silently shed O157 in feces, playing a common role in pathogen epidemiology. Understanding risk factors for cattle O157
shedding is paramount to developing techniques that mitigate spread from dairies to the human food chain. Because these
cattle mount no O157 immune response, and have parallel diets and environmental exposures, we hypothesize that the structure of the colonic microbial community influences a given animal’s risk of becoming colonized with and shedding O157.
The current study aimed to longitudinally describe O157 shedding patterns in early lactation cattle over five days, using gold
standard laboratory techniques. Microbial community differences between shedding and non-shedding individuals were
measured via 16s rRNA Illumina sequencing. Cattle were seen to harbor 3 different pathologic variants of O157: commensal O157, stx O157, and non-stx EaeA O157. Many cattle shed no O157 (4/40) or only commensal O157 (20/40). Those
that shed pathogenic O157 did so either one time (11/40), two times (3/40), three times (1/40) or four times (1/40) over the
course of five days. When cows were classified based on shedding pattern or pathotype, analysis of microbial community alpha and beta diversity metrics revealed no gross differences between categories. However, when individual OTU abundance
was analyzed, members of several genus’ differed by sample category, namely: Ruminoccoccus, Turcibacter, Streptococcus,
Treponema, and Roseburia. Understanding these microbial community changes may eventually guide on-farm strategies
that mitigate O157 dissemination, ultimately protecting the human food chain.
Research Grant: This work was funded in part by USDA NIFA Health and Disease Formula Funds (CRCFY2015
1004896), and an award through MoBio and the Earth Microbiome Project
Student Support: USDA:APHIS:VS:CEAH 15-9200-0443-CA
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The effects of natural and synthetic retinoid therapy on feline squamous cell carcinoma cell lines
Alexa Stephen,Marc Salute, and Carlos Souza
Small Animal Clinical Sciences, University of Florida College of Veterinary Medicine, Gainesville, Florida
Squamous cell carcinoma is the most common oral malignant neoplasia in cats. The existing therapies which include surgery, radiation, and chemotherapy often fail to control this locally invasive tumor and can be associated with significant
morbidity. Retinoids, derivatives of vitamin A, have been shown to inhibit cell proliferation by terminal differentiation and
apoptosis. The current study evaluated two retinoids, all-trans retinoic acid and fenretinide, and their abilities to induce
apoptosis and reduce the production of tumor-promoting cytokines (IL-1, IL-6, IL-8, VEGF and MCP) in vitro. All-trans
retinoic acid induced apoptosis, however not in clinically relevant doses. Fenretinide, a synthetic retinoid, induced apoptosis
and reduced associated cytokines at low micromolar concentrations. Integration of fenretinide with a chemoradiation therapy could provide a synergistic effect, reduce morbidity, and circumvent the limitations of current treatments while providing
higher survival rates. Data acquisition is near completion.
Research Grant: University of Florida College of Veterinary Medicine
Student Support: Florida Veterinary Scholars Program and Merial
Characterization of exosome miR-9 expression in canine osteosarcoma
Olivia A. Stephenson, Joelle M. Fenger
MicroRNAs (miRNAs) are small non-coding RNAs that regulate gene expression and contribute to tumorigenesis. Exosomes are small (40-150 nm) cell membrane derived vesicles that are secreted into bodily fluids such as serum and urine.
These vesicles function in intercellular communication as they transfer their contents, including miRNAs, between cells and
may serve as non-invasive biomarkers in various malignancies. We identified miR-9 as being highly expressed in canine
osteosarcoma (OSA) tumors and OSA cell lines and found that overexpression of miR-9 in normal osteoblasts and OSA cell
lines promotes cell invasion in vitro. We hypothesize that high levels of miR-9 will be detected in exosomes derived from
canine OSA cell lines. We further hypothesize that serum exosome miR-9 will be increased in dogs with OSA compared to
healthy controls and that tumor-associated miR-9 levels will decrease following therapeutic intervention. Exosomes were
isolated from conditioned media from cell lines or patient serum and analyzed using NanoSight imaging. qRTPCR demonstrated that exosome miR-9 expression is higher in canine OSA cells compared to normal osteoblasts. Data obtained from
serum exosomes from healthy dogs and dogs with OSA found that miR-9 transcript is not detectable by qRTPCR; however,
studies are underway to evaluate circulating miR-9 levels in an expanded cohort of dogs. Our findings demonstrate that
miR-9 is detectable in canine OSA cell-derived exosomes and that exosome miR-9 expression is increased in canine OSA
cell lines compared to normal osteoblasts. MiR-9 was not detectable in serum-derived exosomes from healthy dogs or dogs
with OSA, suggesting that circulating miR-9 may not be a useful biomarker for OSA.
Research Grant: Morris Animal Foundation D14CA-057
Student Support: NIH T35 OD010977
271
Use of susceptible and avirulent E. coli to reduce antimicrobial-resistant coliforms in milk-fed calves
Daniel J. Stern, Bo Norby
Michigan State University, Large Animal Clinical Sciences, College of Veterinary Medicine, East Lansing, MI
Antimicrobial resistance is a growing global concern. The use of antimicrobial drugs in food animal production plays an
important role in the accumulation and spread of antimicrobial-resistant bacteria. As regulations tighten on antimicrobial
use, the dairy industry needs to find novel ways to reduce resistant microbial shedding in order to avoid bans on the use of
antibiotics important to human medicine. Our lab conducted a preliminary study that showed a one-log decrease in ceftiofur-resistant coliform bacteria in pre-weaned dairy calves by feeding a mix of antimicrobial-susceptible and avirulent E.
coli strains. In the current study, 12 calves were enrolled. Six calves will be fed the susceptible and avirulent E. coli strains
and the other 6 will serve as controls. The outcomes of interest will be 1) absolute and relative total coliform counts and
coliform bacterial counts resistant to ceftiofur, ampicillin and tetracycline and 2) presence of approximately 200 resistance
genes and the microbiome composition. We expect to find that feeding a cocktail of susceptible and avirulent E. coli to
calves will result in a decrease of resistant intestinal coliform bacteria as compared to the control that will not receive the E.
coli. We also expect to determine how the E. coli cocktail affects the abundance of approximately 200 resistance genes by
multiple singleplex PCR as well as the intestinal microbiome.
Research Grant: Michigan Animal Agriculture Alliance Proposal #AA16-012
Discovery of the causal variant for acquired laryngeal paralysis polyneuropathy in the dog
Allison Stilin, Peter Muir, John Svaren, Lauren Baker, Emily Binversie, Alex Piazza, and Susannah Sample
Department of Surgical Sciences (Stilin, Muir, Baker, Binversie, Piazza, Sample) and Department of Comparative
Biosciences (Svaren), School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI
Idiopathic acquired laryngeal paralysis polyneuropathy (ALPP) is a late onset disease leading to respiratory distress, hindlimb weakness, and death in large breed dogs. Labrador retrievers represent about 65-75% of ALPP cases, and this breed
specificity indicates the strong likelihood of a genetic basis for the disease. Genome-wide association studies (GWAS) are
used to localize the variation(s) in the genome that contributes to the disease by testing single nucleotide polymorphism
(SNP) markers in affected and control dogs to find associations with the disease phenotype. Previous pedigree and phenotype analyses in our laboratory predict ALPP to be an autosomal dominant Mendelian trait. However, the location of the
mutation is currently unknown. The objective of this study was to localize the gene mutation(s) causing ALPP in Labrador
retrievers. SNP markers from 103 affected and control dogs were analyzed using the Illumina Canine HD BeadChip, which
identified potential SNPs on CFAs 6 and 21 that significantly associate with ALPP. These findings will aid in localizing
our region of interest for whole-genome sequencing. The identification of the casual variant will be used to design a PCRbased genetic test and will enhance our understanding of the progression and neuropathologic features of ALPP, critical for
development of future therapies. This work may also provide a naturally occurring animal model for human medicine, as
ALPP is clinically similar to peripheral neuropathies in humans.
Research Grant: National Institutes of Health; UW-Madison School of Veterinary Medicine Companion Animal Grant;
American College of Veterinary Surgeons
Student Support: UW-Madison School of Veterinary Medicine
272
The effects of human interactions on gastrointestinal parasites in Mediterranean mouflon in southern France
Suzanna M Storms, Eloise Barge, Marie-Therese Poirel, Slimania Benabed, Matheiu Garel, Pascal Marchand,
Christian Itty, Gilles Bourgoin
Laboratoire de Parasitologie Veterinaire (Storms, Barge, Poirel, Benabed, Bourgoin), VetAgro Sup, Universite de Lyon,
Marcy l’Etoile, France Office National de la Chasse et de la Faune Sauvage (Marchand), Juvignac, France
Laboratoire d’Ecologie Alpine (Garel, Marchand), Universite de Savoie, Le Bourget-du-Lac, France
Human interaction and encroachment is a major stressor in wild animals. Previous studies in our lab have shown that hunting and touristic activities cause disturbances in movement and grazing patterns in Mediterranean mouflon (Ovis gmelini
musimon x Ovis sp.). The purpose of this study was to determine if there is a correlation between gastrointestinal nematode
levels and the amount of human disturbance in Mediterranean mouflon in the Caroux-Espinouse massif in southern France.
Fecal samples were collected from three adjacent wildlife refuges during different periods of 2015 and 2016. These sampling periods were corresponded with high and low hunting seasons (September- February, peak November-February) and
tourist seasons (March-October, peak March-August). Samples were collected and sent to the lab to be analyzed. Fecal egg
counts were performed first, using a MacMaster technique with 5g of fecal material. Coproculture was performed next,
with an incubation at 25°C for 10-12 days. The samples were humidified every other day to prevent dryness. The Baermann
technique was used to collect the larvae from the incubated samples. 100 L3 larvae from each sample were then identified in
order to find the proportion of gastrointestinal nematodes present during each hunting and tourist season in the three adjacent
zones. Preliminary data analysis from fecal egg counts and stage 3 larvae suggests a tendency toward lower GI nematode
levels in the less touristic areas. Forthcoming data analysis will provide a clearer understanding of the interaction of the GI
parasites and the high and low hunting and tourist seasons.
Transient Receptor Potential Melastatin 4 channel enhances osteogenesis of rat dental pulp stem cells
Kelsie Stovall, Tran Doan Ngoc Tran, Yawen Hu, Tanyawan Suantawee, Shaomian Yao, Sirichai Adisakwattana
and Henrique Cheng
Department of Comparative Biomedical Sciences, Louisiana State University SVM, Baton Rouge, LA (Yao, Cheng),
Program in Biomedical Sciences, Graduate School, Chulalongkorn University, Bangkok, Thailand (Suantawee,
Adisakwattana)
Dental pulp stem cells (DPSCs) are multipotent cells that can differentiate into osteoblasts and adipocytes, and have the potential to be used in tissue regeneration. However, the mechanism controlling the differentiation process is not fully understood. Previously, we identified two members of the Transient Receptor Potential Melastatin (TRPM4 and 7) ion channels
in rat DPSCs and demonstrated that they are required for cell proliferation. In this study, we examined for the first time their
role in osteogenic and adipogenic differentiation. We utilized RT-PCR to identify TRPM4 and 7 gene expression and the
patch-clamp technique to determine whether these channels were functionally active in rat DPSCs. To investigate their role
in differentiation, we utilized specific TRPM4 (9-phenanthrol) and TRPM7 (carvacrol) channel blockers during the differentiation process. Osteoblast and adipocyte formation were quantified by Alizarin Red S and Oil Red O staining. To elucidate
the mechanism by which these channels control the differentiation process, we performed real-time Ca2+ imaging analysis
during channel suppression and ATP stimulation to induce intracellular Ca2+ signals. Inhibition of TRPM4 enhanced osteogenesis more than TRPM7 suppression. Nevertheless, rat DPSCs failed to differentiate into adipocytes. Furthermore, inhibition of TRPM4 transformed the ATP-induced biphasic Ca2+ signaling pattern into sustained Ca2+ oscillations. These results
suggest that TRPM4 may be used as a potential target to promote bone regeneration using stem cell therapy.
Student Support: NIH T35 Training Grant 5T35OD011151-13, LSU SVM Summer Scholars Program
273
A systematic review of the literature to identify and quantify host and vector competence and abundance of JEV
Erin Strathe, Ana Rute Da Silva Oliveira, Luciana Etcheverry Hernandez, Lee Cohnstaedt, Scott McVey,
Natalia Cernicchiaro.
Department of Clinical Sciences (Strathe); Department of Diagnostic Medicine and Pathobiology (Oliveira, Hernandez,
Cernicchiaro), College of Veterinary Medicine, Kansas State University; USDA-ARS Arthropod-Borne Animal Diseases
Research (Cohnstaedt, McVey), Manhattan KS
Japanese Encephalitis virus (JEV) is a mosquito-borne arbovirus that causes endemic and epidemic encephalitis in Eastern
and Southeastern Asia. Swine and wading birds serve as reservoirs for the virus, which can be transmitted to humans via
mosquitos. Prevention is highly important, due to a lack of specific treatment options. The potential risk of inadvertent introduction of JEV into United States is a concern because of the public health, trade and economic implications of entry of an
emerging disease. Our objective was to gather and appraise original scientific literature to identify and quantify competence
and abundance of potential hosts and vectors of JEV in United States and worldwide using a systematic review of the literature. Our study began by posing a research question and selecting key words for searching abstracts in electronic databases.
The title and abstract of the identified publications were then screened for relevance utilizing a defined set of exclusion and
inclusion criteria. After subjecting the abstracts to two rounds of relevance screening, data were extracted from the relevant
full-text articles and their risk of bias assessed. These data will then be analyzed quantitatively using statistical methods
(meta-analysis). A total of 1405 abstracts were identified in the original search from electronic databases and grey literature.
The initial round of relevance screening resulted in the identification of 568 relevant abstracts. A second screening narrowed
the papers from which data was to be extracted to 290. The data obtained from this study will be utilized to populate vector
and host entry pathways of a risk analysis to evaluate risks associated with introduction of JEV into the United States.
Research Grant: United States Department of Agriculture, Agricultural Research Service (USDA-ARS) Project No:
5430-32000-008-05S.
Student Support: Merial.
A questionnaire-based study on predisposing and risk factors for gastric dilatation volvulus in dogs
Shayna Streu, Laura Nelson, Grace Lai, and Heather Defore
Michigan State University, East Lansing, MI
Gastric dilatation volvulus (GDV) is a life-threatening acute condition that has been deemed as multimodal. As researchers
continue to investigate what these causes may be, some have discovered risk factors that appear to increase the chances of
an at-risk dog to developing GDV, some of which include having an anxious disposition and eating fewer meals throughout
the day. Using the data compiled on questionnaire forms given to owners of dogs that had been affected by GDV and owners
whose dogs were not affected by the condition but were a breed at higher risk of developing GDV, 25 affected dogs and 52
not affected dogs were evaluated. From those groups, 10 affected Great Danes were compared against 14 not affected Great
Danes, and then against 15 affected dogs that were identified as any other breed. Results of analysis of these groups for any
differences indicated that, when all affected and all not affected were compared, there were significant differences in age (p
= 0.016), body condition score (p = 0.042), vomiting frequency (p = 0.002), anxiety (p = 0.020), adaptability (p = 0.020),
thoracic circumference (p = 0.010), and height (p = 0.002). When affected Great Danes were compared to not affected Great
Danes, significant differences found were vomiting frequency (p = 0.024), anxiety (p = 0.033), fear (p = 0.014), and adaptability (p = 0.003). Finally, when affected Great Danes were compared against other affected breeds, a significant difference
was found that indicated Great Danes with raised bowls were more likely to bloat than other breeds (p < 0.001). The advent
of this data helps reinforce previous studies and offers new variables not previously explored, offering expansion for future
studies.
Research Grant: American Kennel Club
274
Effects of caffeine on injured and uninjured intervertebral discs in a whole organ culture model
Olivia E. Stricklin, James T. Stannard, Benjamin T. Raines, Aaron M. Stoker, and James L. Cook
University of Missouri
Intervertebral disc (IVD) disorders resulting in pain are prevalent, but the mechanisms of IVD degeneration are not fully
understood. A recent study demonstrated decreased cell viability and glycosaminoglycan (GAG) content in injured rat tail
IVDs within 7 days of nicotine exposure. Caffeine has similar traits to nicotine and is widely consumed. We therefore hypothesize that caffeine will be associated with a marked loss of cell viability of injured IVD explants compared to control
and uninjured explants. IVDs collected from Sprague Dawley rat tails were randomly assigned to one of four groups with
or without injury: control (CON), low (5 mg/L) (LCAF), medium (10 mg/L) (MCAF), or high caffeine (15 mg/L) (HCAF).
Explants were cultured for 21 days in a rotating wall vessel bioreactor. Culture media was refreshed and tissue was harvested
every 7 days. Disc cell viability was assessed using fluorescent microscopy, and GAG and collagen content were determined
using a DMMB and hydroxyproline assay, respectively. Caffeine exposure was associated with an inverse, dose-dependent relationship where cell viability decreased as caffeine concentrations increased. Uninjured IVD cell viability analysis
demonstrated significantly higher cell viability in LCAF compared to MCAF and HCAF at day 21. Uninjured MCAF and
HCAF cell viability was significantly lower at day 21 compared to CON. Injured HCAF cell viability was significantly
lower at days 7 and 14 compared to uninjured HCAF. Biochemical analyses of uninjured IVDs demonstrated a downward
trend in proteoglycan to collagen ratio for LCAF, suggesting a potential detrimental effect of caffeine on IVD extracellular
matrix composition. Therefore, caffeine may play a role in back pain.
Research Grant: UMOA Grant
Genotype and phenotype of sudden acquired retinal degeneration syndrome (SARDS) in dachshunds
Stephanie Stromberg, Sara Thomasy, Ann Cooper, Ariana Marangakis, David Maggs, and Danika Bannasch
Department of Surgical & Radiological Sciences (Stromberg, Thomasy, Marangakis, Maggs), Veterinary Medical
Teaching Hospital (Cooper), Department of Population Health and Reproduction (Bannasch), School of Veterinary
Medicine, University of California Davis, Davis, CA
Sudden acquired retinal degeneration syndrome (SARDS) is a common cause of incurable blindness in dogs. This disease is
diagnosed by a rapid loss of vision combined with a flatline electroretinogram (ERG) response and a normal appearing fundus. The cause of this disease is unknown, although several hypotheses have been suggested, and one criticism of SARDS
research so far is the lack of a clear definition of the disease. To better characterize the disease in vivo, we are using optical
coherence tomography (OCT) to measure and compare the thickness of retinal layers in six dogs affected with SARDS for <
1 year, six dogs affected with SARDS for > 1 year, and twelve healthy control dogs. We hypothesize that affected dogs will
have decreased thickness of the retinal photoreceptor layer at < 1 year, and decreased overall retinal thickness at > 1 year. To
date, six dogs have undergone OCT imaging with more scheduled. In addition, discovery of a genetic component of SARDS
in Dachshunds (one of the most commonly affected breeds) could lead to a deeper understanding of the disease’s etiology,
identification of at-risk animals, and development of treatments. We hypothesize that we will be able to identify regions of
the canine genome that are associated with the presence of SARDS in a genome-wide association study (GWAS) using a
new Affymetrix canine SNP array performed on 24 affected and 24 control dachshunds. Thirty-six samples have undergone
GWAS and are awaiting data analysis. Of those, 3 did not produce results, most likely due to poor DNA quantity or purity.
A further 12 samples will be analyzed once the array is commercially available.
Research Grant: UC Davis Center for Companion Animal Health, Vision for Animals Foundation, and NIH grants K08
EY021142 and P30 EY12576
275
Alyssa A. Strumpf, Valerie Johnson, Renata Impastato, Steven Dow
Center for Immune and Regenerative Medicine, Department of Clinical Sciences, Colorado State University,
Fort Collins, CO
A frequent complication of diabetes is chronically infected foot ulcers with multi-drug resistant bacteria. To explore new
approaches for treatment of these wounds, our group investigated the use of activated mesenchymal stem cells (MSC). Previous studies conducted in murine and canine models show that administration of activated MSC exert antimicrobial properties and serve as an interface between infection and activation of the innate immune system. To advance the use of MSC
therapy in humans, we conducted in vitro studies examining the antimicrobial properties of human MSC and the interaction
of MSC with human innate immune cells. We hypothesized that human MSC could secrete antimicrobial factors that would
further increase the effectiveness of antibiotics for killing of drug-resistant bacteria and also could upregulate cells of the
innate immune system. In vitro killing assays with susceptible and drug-resistant strains of S. aureus were used to evaluate
direct antimicrobial activity, while co-cultures with neutrophils or monocytes and conditioned media were used to assess
innate immune cell interactions. Our studies revealed that MSC exerted direct bacterial killing activity against susceptible
and drug-resistant strains of S. aureus, and furthermore, significantly enhanced the effectiveness of antibiotics. Moreover,
conditioned medium from MSC triggered significant upregulation of neutrophil phagocytosis and stimulated monocyte migration. These findings confirm the antimicrobial properties of human MSC and suggest important interactions with innate
immune cells. These results, combined with data from previous studies, will help advance the translation of antimicrobial
stem cell therapy to human clinical trials.
Research Grant: Grubstake Fund (UC Denver and Gates Center for Regenerative Medicine) and Shipley Foundation
Program in Applied Regenerative Medicine
Deciphering the interactions between feline mesenchymal stem cells (MSCs) and CD8+ T cells in vitro
Ash Sundaram, Naomi J. Walker, Dori L. Borjesson
UC Davis School of Veterinary Medicine, Davis, CA
Mesenchymal stem cells (MSCs) are multipotent stem cells with the ability to modulate both innate and adaptive immune
responses. Hence, they are a promising therapy for both immune mediated and inflammatory disorders. Our lab investigates the use of adipose derived MSCs for treatment of feline chronic gingivostomatitis (FGGS), which is a severe and
debilitating immune mediated oral disease of cats. This can serve as a spontaneous disease model for immune mediated
oral mucosal diseases in humans including oral lichen planus, pemphigus and apthous stomatitis. The diseases in cats and
humans are both characterized by T cell activation and tissue destruction, and both blood and oral tissues from human and
feline patients have shown increased percentages of CD8+ T cells and effector T cells. Recent research in our lab and clinical
trials at the UC Davis Veterinary teaching hospital show that in cats who have not responded to conventional therapy, MSC
therapy is safe and over 70 % effective. But the mechanism by which MSCs reduce inflammation is not well understood. We
are conducting series of in vitro co-incubation experiments of CD8+ T cells and MSCs followed by MSC primed CD8+ T
cells and CD4+ T cells to decipher this interaction between feline MSCs and CD8+ T cells that reduces inflammation. Our
experiments help us understand two things: 1) If MSC priming or reprogramming of CD8+ T cells results in a population of
CD8+ T Regs with immune suppressive or regulatory function. We evaluate this by their ability to suppress activated CD4
T cells. 2) If MSC priming or reprogramming of CD8+ T cells is mediated by cell to cell contact or soluble mediators. We
will present the results of these experiments in the poster.
Research Grant: NIH Grant IR21DE024711-01
Student Support: STAR NIH Grant
276
Defining the relationship between female sex hormones and myocardial fibrosis in aortic-banded mini-swine
Tracy Swanson, Jessica A. Hiemstra, T. Dylan Olver, Jenna C. Edwards, Madeleine Dionne, Pamela K. Thorne,
Jan R. Ivey, Craig A. Emter
University of Missouri-Columbia, College of Veterinary Medicine, Department of Biomedical Sciences
Approximately 50% of heart failure (HF) cases in the United States are diagnosed as heart failure with preserved ejection
fraction (HFpEF). HFpEF is diagnosed in women twice as often as in men, with morbidity and mortality rates increasing
with age. These sex and age-specific effects point to the involvement of menopause, during which aging women naturally
lose ovarian sex hormone production. The purpose of this study was to determine the relationship between HFpEF and
ovarian sex hormones on the development of left ventricular (LV) fibrosis in response to chronic pressure-overload. For
this study, an ovariectomy (OVX) model of menopause and aortic-banded (AB) model of HFpEF were used in female mini
swine divided into four groups: control-intact (CON, n=7), AB-intact (AB, n=7), control-ovariectomized (CON-OVX, n=6),
and AB-ovariectomized (AB-OVX, n=7). Total LV collagen protein and mRNA levels of specific collagen (Collagen I &
III), matrix metalloproteinase (MMP-2 & -9), and tissue inhibitors of MMP (TIMP-1 & -4) isoforms were measured. Tissue
samples from the LV were hydrolyzed in hydrochloric acid and collagen-specific hydroxyproline was quantified using a colorimetric assay and reflected as the ratio of total collagen: total protein. Quantitative RT-PCR was used to measure mRNA
levels of extracellular matrix components and their regulatory biomarkers. We hypothesize that AB pigs will have increased
total collagen and increased MMP-2, MMP-9, TIMP-1, TIMP-4, Collagen I, and Collagen II mRNA levels when compared
to CON. These pathological changes will be exacerbated by the loss of ovarian sex hormones.
Research Grant: NIH/NHLBI R01 HL112998 (CAE)
Student Support: IDEXX-BioResearch
Vaccination of small ruminants against Haemonchus contortus using fecal antigens
Connor T. Tageant, Vicky E. Kelly, Rachel M. Taupier, Philip H. Elzer, and James E. Miller
School of Animal Science, College of Agriculture (Tageant, Elzer, Miller) and Department of Pathobiological Science,
School of Veterinary Medicine (Tageant, Kelly, Taupier, Elzer, Miller), Louisiana State University, Baton Rouge, LA
Gastrointestinal parasites, especially nematodes such as Haemonchus contortus, are a major problem in the production of
small ruminants. The anthelmintics which were traditionally used to treat for Haemonchus have lost their efficacy. Various
management strategies have been developed as alternatives for controlling the parasite burden in small ruminants. One of
these strategies is vaccination. Currently vaccines are created using antigen taken from adult nematodes that are extracted
form heavily parasitized slaughtered animals. This is a very costly process. This project is attempting to use the Haemonchus antigen that is taken from feces. Fecal samples were harvested form heavily parasitized animals. The samples were
mixed with phosphate buffered saline and antibacterial/antimycotic to make a solution. The solution was then filtered and
centrifuged to remove particulate. The protein in the solution was precipitated using ammonium sulfate. A chromogenic
assay was used to test for endotoxin and then endotoxin was then removed using a column removal kit. An SDS-PAGE gel
was used to determine the presence of antigen. The antigen will then be extracted and used in a vaccination trial.
277
The effects of inflammation on the pharmacokinetics of flunixin in steers using in vivo ultrafiltration
David Tamas, Alex Taylor, Derek Foster, Liliana Carbajal, Kristen Messenger
North Carolina State University CVM, Raleigh, NC
Knowledge of active drug concentrations and effects directly at sites of action is critical to understanding pharmacokinetic-pharmacodynamic (PK/PD) targets for nonsteroidal anti-inflammatory drugs (NSAIDs). Flunixin (FX) is an approved
NSAID in cattle, but flunixin PK/PD have only been reported using artificial tissue cage models of inflammation. The purpose of this study was to describe the PK of FX in plasma and interstitial fluid (ISF) of steers using a novel incision model
of inflammation. We hypothesized that protein-unbound flunixin drug concentrations would be greater in inflamed tissues
compared to control (uninflamed) tissues, and that flunixin concentrations in ISF would be detected longer than concentrations found in the plasma. Six healthy Holstein steers received 2.2 mg/kg FX meglumine intravenously every 24 hours for
2 days. Ultrafiltration probes were placed in a laparotomy incision (inflamed) and between the scapulae (control). Blood
and ISF samples were obtained at pre-determined time points over 144 hours. FX was quantified using UPLC-MS. PK
parameters of FX were estimated using compartmental and non-compartmental methods, with a naive pooled approach for
ISF data. RESULTS: Geometric mean (CV%) estimates for the volume of distribution, and clearance, for FX in plasma
were 0.89 (25.3)L/kg, 179.7 (43.4)mL/hr/kg, and 6.0 (37.0)hrs, respectively. Noncompartmental estimates for area under
the curve in control ISF was 128.7 ng/mL*hr, and was 586.7 ng/mL*hr in inflamed ISF. Our plasma results agree with published PK estimates in cattle, and the ISF results suggest that protein-unbound FX exposure is greater in inflamed tissue fluid
compared to uninflamed tissue fluid.
Research Grant: Internal funds from the Comparative Pharmacology and Anesthesiology Laboratory
Student Support: Veterinary Scholars Program and Fund for Discovery
Karena Tang, Wael Khamas, Fashid M. Shahriar, and Gagandeep Kaur
Western University of Health Sciences, CVM, Pomona, CA
The amygdala is a cluster of about 13 structurally diverse nuclei in the temporal lobe that is an important control center
for emotions such as fear and aggression. While structural and morphometric studies have been performed mainly in rats
and mice, few have focused on other species. In this study, we characterized the neuronal morphology and cytoarchitecture
of amygdala in 3 domestic cats from 1 to 10 years of age. Post mortem brain tissue samples of the right amygdala were
trimmed, fixed, and sectioned into 30 mm serial sections using a cryostat. Every 11th and 12th sections were mounted on
superfrost plus slides and stained with cresyl violet to confirm nuclei localization, study neuronal architecture, and measure
the rostrocaudal extent of the amygdala. The results showed that the rostrocaudal length of the amygdala ranged from 6.7 7.3 mm in cats. The average neuronal diameter was 10.64 mm in the lateral nucleus, 14.63 mm in the medial nucleus, 11.37
mm in the central nucleus, and 11.93 mm in the basal nucleus, which have specific locations within the amygdala. We will
include more dimensions of the amygdala and compare these results with cats older than 10 years in future studies. These
anatomical findings may complement physiological findings to pave the way for further research in feline behavior and
comparative studies to improve feline health and welfare.
Research Grant: Intramural WesternU
Student Support: WesternU Summer Research Fellowship from the office of the VP of Research and Biotechnology
278
Aedes Aegypti mosquito transmission and the development of STAT2 KO hamster animal model for Zika virus
Cassandra Tang-Wing, Alexis Dadelahi, Jeffrey Adamovicz
School of Veterinary Medicine, St. George’s University (Tang-Wing), Department of Veterinary Pathobiology,
College of Veterinary Medicine, University of Missouri (Dadelahi, Adamovicz)
The mosquito-borne Zika virus (ZIKV) responsible for the recent outbreak of febrile illness is a recent subject undergoing
intense study. The virus appears to be asymptomatic in many cases, yet the neurological implications, resulting in microcephaly and other aberrant neurological developments in neonates as well as associated Guillan- Barre syndrome in select
individuals, have raised concern regarding the spread of the virus. Information concerning disease transmission, immune
response to infection and efficacy of antiviral components in early gestation remain unclear. In order to address these
unknowns an appropriate and effective animal model is necessary to study the pathogenesis and pathology of the virus.
Previous studies have used immunocompetent mice, deficient in interferon a/b and g receptors (A129 and AG129 mice),
however, they do not directly mimic the pathogenicity in humans nor demonstrate the natural route of infection through
mosquito bites. Therefore, the aim of the current study is to develop an animal model and study mosquito transmission
and immune response of ZIKV that appropriately represents human infection. Aedes Aegypti mosquitoes were fed on vero
cell viral stocks and viable virus was detected in midgut and bodies, demonstrating their ability to be competent vectors.
These mosquitoes will be exposed to STAT2 KO hamsters to mimic natural infection. Viral titers and clinical signs will be
analyzed, and liver, spleen, neurological tissue, and testes/ovaries will be collected for histopathology and immunohistochemistry. By identifying the viral-induced infectivity and pathology in the STAT2 KO hamsters, we expect that this will
be a suitable animal model to further support other ZIKV studies.
Student Support: University of Missouri, College of Veterinary Medicine
Diagnostic accuracy of Keto-Test milk strips for cowside detection of elevated milk b-hydroxybutyrate
Stephanie J. Tarlowe, Kathryn D. Bach, Jessica A. A. McArt
Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University,
Ithaca, NY
Hyperketonemia is a common disease of early postpartum dairy cattle characterized by an excess of circulating blood ketone
bodies with an absence of clinical signs. This disorder leads to economic losses from decreased milk production, reduced
reproductive performance, and increased risk of displaced abomasum and removal from the herd. Ketones can be detected
in blood, urine, and milk, with the gold standard being laboratory measurement of blood b-hydroxybutyrate (BHB). While
cowside blood tests provide an accurate means of diagnosis, this method is time-consuming and invasive; milk collection
does not pose these same challenges. Keto-Test milk dip-strips provide an easy, non-invasive means of testing milk BHB
concentration cowside, but more research is needed in order to determine the diagnostic accuracy of these strips and establish definitive cut-points for elevated milk BHB based on the color change of the strip. Our objective is to determine the diagnostic accuracy and precision of Keto-Test dip-strips in measuring milk BHB compared to the gold standard mid-infrared
(IR) spectroscopy method. Milk was collected on 2 herds from 50 dairy cows between 2 and 16 days in milk using proportional samplers. Samples were tested immediately with the Keto-Test dip-strip according to label directions and then transported on ice to the laboratory for testing via IR methodology. The two diagnostic methods will be compared using a kappa
calculation for inter-method agreement. Precision of the Keto-Test dip-strips will be determined by repeatedly measuring 1
milk sample in each of the 4 colored BHB concentration categories using 10 different test strips. Results are pending.
Research Grant: USDA National Institute of Food and Agriculture, Hatch project NYC-478432; Elanco Animal Health
279
Marek’s disease infectivity in genetically susceptible and resistant chickens
Melissa Tatro, Osvaldo Anacleto, Andrea Doeschl-Wilson, Hans Cheng, and John Dunn
College of Veterinary Medicine, Michigan State University, East Lansing, MI (Tatro), Roslin Institute and R(D)SVS,
University of Edinburgh, Midlothian, Scotland (Anacleto, Doeschl-Wilson), USDA ARS Avian Disease and Oncology
Laboratory (Cheng, Dunn)
Marek’s disease is a viral disease of chickens causing tumor growth and neurological signs leading to rapid death. While
vaccines exist for this economically important disease, current control efforts are problematic. Vaccines prevent tumor development but do not prevent viral shedding. Additionally, the virus is transmitted by feather dust; therefore, it is ubiquitous
in the environment. This presents serious potential for the virus to evolve to the point that vaccination programs are no longer effective. However, some genetic lines of chickens have been shown to be more resistant to Marek’s disease than others.
We hypothesized that these genetically resistant chickens also shed less virus and are less infective than their genetically
susceptible counterparts. In order to test this hypothesis, we first needed to define the infectivity period, which was the focus
of this study. To do this, we used genetic lines with varying susceptibility. Viral donor birds were infected at hatch and transplanted every 4 days into groups of susceptible recipient birds. Then, viremia of both donors and recipients was measured
via qPCR of blood and feather samples. At the end of eight weeks, both donors and recipients were necropsied to look for
pathologic evidence of Marek’s disease. Days 20-28 post-exposure appear to be when infectivity is greatest, though birds
can become infective as early as 12 days post-exposure. Birds exposed to virus less than 12 days after exposure tend to not
develop disease. Resistant birds demonstrated less viral load overall when compared to susceptible ones. This supports the
hypothesis that resistant birds are less infective.
Research Grant: Agriculture and Food Research Initiative Competitive Grant no. 2016-67015-24914 from the USDA
National Institute of Food and Agriculture
Student Support: Joan E. and Richard Witter Scholarship Fund at Michigan State University
Effects of Bedoukian compound concentrations on gastrointestinal nematode development in sheep
Rachel M. Taupier, Connor T. Teagent, Vicky E. Kelly, and James E. Miller
Gastrointestinal nematodes have a large financial and health impact for small ruminant farmers. Recent years has seen an
increase in anthelminthic resistance, and so there is a drive towards finding alternative control methods. This experiment is
a continuation of an in vitro study that used ketone compounds created by Bedoukain Research to reduce fecal egg survival rates. Our goal is to observe the compound’s effects on egg survival within the feces of lambs with naturally occurring
gastrointestinal nematode infections. Five treatment groups of lambs were treated with differing doses of Bedoukain’s compound to determine if higher concentrations of the compound will have an increasing impact on the infection. Additionally,
we explored whether steady doses over an extended period of time would increase the compound’s effect. If these treatments
have a statistically significant effect, it will open the door for development of this compound in practical applications.
Research Grant: USDA 1433 Grant Funds and Bedoukian Research
Student Support: USDA 1433 Grant Funds and Bedoukian Research
280
Control of lupus nephritis by manipulating gut microbiota during active disease
Vincent Tavella, Qinghui Mu, Ansar Ahmed, Xin Luo
Department of Biomedical Sciences and Pathobiology, College of Veterinary Medicine, Virginia Tech, Blacksburg, VA
Systemic Lupus Erythematosus is an autoimmune disorder characterized by severe and persistent inflammation that causes
damage in multiple organs. Disease susceptibility is influenced by genetic, immunological, hormonal, and environmental
factors which result in a breakdown in tolerance towards self-antigens. The dynamics of gut microbiota in lupus-prone
MRL/lpr mice have been described and suggest a potentially important role of microbiota on lupus pathogenesis. A recent
study from our lab has shown that increasing beneficial bacteria such as Lactobacillus spp. in the gut prior to disease onset
can attenuate lupus nephritis in these mice. Here we tested the hypothesis that alteration of the gut microbiota after disease
development can ameliorate lupus nephritis and serve as a potential treatment to attenuate active disease. Weekly oral gavage of Lactobacillus spp. was administered to MRL/lpr mice starting 9-10 weeks of age whereon disease manifestations
developed. All Lactobacillus strains, including oris, rhamnosus, reuteri, johnsonii and gasseri were cultured every week,
mixed, and inoculated at 2x108 cfu/strain per dose. As vancomycin is known to enrich Lactobacillus spp. in the gut microbiota, we administered the antibiotic in drinking water (1 g/L) as a separate experimental group. Urine was collected
weekly for measurement of proteinuria. Feces were collected weekly to extract bacterial DNA that was sequenced for microbiota characterization. Blood samples were taken on week 15 to assess auto-antibody levels. In final dissection, kidney
was collected for pathological analysis of lupus nephritis. Our results suggest that treatments with both Lactobacillus and
vancomycin during active disease are beneficial.
Research Grant: Merial Veterinary Scholar Program & Virginia-Maryland College of Veterinary Medicine
Evaluation of health in Texas tortoises via multiple patient-side analytes
Michelle C. Taylor, and J. Jill Heatley
Department of Small Animal Clinical Sciences (Heatley), Veterinary Student (Taylor) College of Veterinary Medicine and
Biomedical Sciences, Texas A&M University, College Station, TX. and Schubot Exotic Bird Health Center (Heatley)
The Texas tortoise, Gopherus berlandieri, is the only native tortoise species in Texas and is considered threatened. Little is
known regarding reptilian inflammatory response and markers of overall health. Mycoplasma spp. and Tortoise Herpesvirus
2 (TeHV2) exposed Texas Tortoises (n= 10) and apparently healthy male Texas Tortoises (n = 10) maintained at the Texas
A&M University, College of Veterinary Medicine and Biomedical Sciences, Winnie Carter Exotic Animal Center were
sampled via jugular venipuncture to determine multiple analytes for assessment of tortoise health. Whole blood was used
to determine ESR, fibrinogen, lactate, packed cell volume and total solids via patient side methodologies while lactic acid,
total protein, albumin, globulin (indirectly), uric acid and serum iron were determined from plasma using a standard laboratory analyzer. While plasma fibrinogen was significantly different based on health status the unexpected lesser values found
during determination of fibrinogen in this species suggest that in contrast to birds and mammals, Texas tortoise refractometric index of plasma may decrease in response to heating and centrifugation. In Mycoplasma and TeHV exposed tortoises,
ESR and lactic aide were higher while plasma fibrinogen was lower than in apparently healthy tortoises. Fair agreement of
lactate and lactic acid, as well as total solids and total protein suggests that patient side methods (lactometer and refractometer) are acceptable for clinical use in these species.
281
How does Toxoplasma gondii deliver effector proteins that control host cell functions
Elizabeth F. Tenborg, John C. Boothroyd, and Michael W. Panas
School of Veterinary Medicine, University of California, Davis, CA (Tenborg) Department of Comparative Medicine
(Tenborg) Stanford University, Stanford, CA. Department of Microbiology and Immunology (Boothroyd, Panas),
Stanford University, Stanford, CA.
Toxoplasma gondii is an intracellular eukaryotic parasite that resides within a unique parasitophorous vacuole (PV) inside
a host cell. This location gives the Apicomplexan parasite a distinct advantage in evading detection by the immune system,
yet poses a tangible disadvantage in that the protective PV membrane is also a physical barrier to the introduction of parasite effectors into the host cell, such factors being necessary to modulate host cell function. The mechanism of transit of
these effector proteins across the PV membrane is currently unknown. Our laboratory has identified three T. gondii proteins
(MYR1, MYR2, and MYR3) as candidate components of a pore machinery spanning the PV membrane, with each being
necessary for the translocation of protein effectors from the PV into the host cell. Most recently, we have identified a new
mutant parasite (MFM1.15) exhibiting reduced control of host cell functions, and have subsequently shown this reduced
control is a result of impaired translocation of effector proteins across the PV membrane. Thus, we hypothesize this phenotype is the result of a defect in a component of the translocation machinery. By sequencing the three known loci (MYR1/2/3)
and finding that all three are fully wild type, we can conclude this MFM1.15 mutant contains a mutation in another, yet undiscovered, piece of the translocation machinery. A current whole-genome analysis is underway to detect the precise genetic
disruption within this variant, furthering our understanding of how T. gondii delivers proteins to co-opt host cell functions.
Research Grant: NIH R21 AI112962-01
Student Support: NIH Office of Director, Division of Comparative Medicine
Evaluation of refractometry to detect failed transfer of passive immunity in pre-weaned beef calves
Alexis C. Thompson, Liesel G. Schneider, Min Wang, David R. Smith
Department of Pathobiology and Population Medicine, College of Veterinary Medicine, Mississippi State University,
Starkville, MS
Failed transfer of passive immunity (FTPI) is associated with increased morbidity and mortality in pre-weaned calves. The
gold standard for detecting passive transfer is radial immunodiffusion (RID). An alternative is refractometry which indirectly measures immunoglobulins by estimating the concentration of serum total protein (STP). The objective of this study
was to measure the agreement and establish a cut-off point for FTPI between STP and RID and evaluate other refractometry
scales. Serum IgG concentration from 818 calves from 4 farms was measured by RID and refractometry using scales for
Brix%, specific gravity (SG), and STP. The correlation coefficient between the refractometry scales and the RID IgG concentration were calculated. Sensitivity and specificity of STP was calculated for each 0.1 g/dL increase in STP to determine
an optimal cut-off using an RID value of 1,000 mg/dL IgG to define FTPI. Based on RID, the prevalence of FTPI in beef
calves in our study population was 4.8% (95%CI=4.0%, 6.3%). The association between Brix% and RID, SG and RID,
and STP and RID were significant (P<0.01, r=0.86, 0.85, 0.84, respectively). Based on ROC curve, an optimal cut-off for
STP of 5.9 g/dL to classify calves to FTPI status was identified. At this cut-off the test performance of STP measured by
refractometry was 93% specific (95%CI=91%, 95%) and 95% sensitive (95%CI=83%, 99%). This level of test performance
makes refractometry diagnostically useful to identify calves with FTPI in populations with a high prevalence of calves with
FTPI (e.g. 40%). However, as a screening tool in low prevalence populations STP refractometry the test has a low positive
predictive value and will over-estimate prevalence of FPTI.
Research Grant: Mikell and Mary Cheek Hall Davis Endowment for Beef Cattle Health and Reproduction
282
Mary Timonin, Cansu Agca, Yuksel Agca
Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon SK Canada (Timonin),
Department of Veterinary Pathobiology, College of Veterinary Medicine, University of Missouri, Columbia, MO
Cryopreservation of sperm provides a valuable means of maintaining transgenic mouse strains used in biomedical research
while minimizing the potential for genetic drift or spontaneous loss of phenotype. Sperm is easy and inexpensive to collect, store, and transport between research institutes, but is also sensitive to freezing. The quantity and quality of fresh
sperm influence cryosurvival, with significant variation between mouse lines. As a result, researchers continue to search for
modifications to freezing protocols that can be applied across mouse lines and provide consistent improvements in sperm
survival and function following freezing. Previous research indicates that iodixanol has cryoprotectant properties for sperm,
but its effects on the sperm of mice has not been determined. We hypothesized that the addition of iodixanol to a standard
raffinose-skim milk freezing solution would improve survival and viability of mouse sperm following freezing and thawing.
We collected sperm from the cauda epididymis of 129/SV and FVB/NJ mice. Sperm was mixed with a freezing solution
containing either 0, 5, 15 or 20% iodixanol and frozen in liquid nitrogen. We evaluated viability of thawed sperm by measuring motility, plasma and acrosomal membrane integrity, and mitochondrial membrane potential. More sperm from FVB/
NJ mice frozen with the solution containing 15% iodixanol survived freezing and showed increased progressive motility,
and acrosomal membrane integrity compared to the control and 5% iodixanol freezing solutions. Sperm from 129/SV mice
showed a similar trend. Thus, iodixanol has cryoprotectant properties for sperm from mice of different inbred strains and
may be useful in improving mouse sperm cryosurvival.
Research Grant: NIH U42OD010918-18
Student Support: An endowment established by IDEXX-BioResearch
Development of less invasive functional assessments of malignant hyperthermia (MH) genotypes
Florian Touitou, Gennady Cherednychenko, Shane Antrobus and Isaac N. Pessah
Department of Molecular Biosciences (Touitou, Cherednychenko, Antrobus, Pessah), School of Veterinary Medicine,
University of California, Davis, United States of America; Ecole Nationale Veterinaire de Toulouse (Touitou), Toulouse,
France
MH susceptibility has to be tested in every patient with a familial history or suspected risk factors for this potentially lethal
disorder before they can undergo general anesthesia with gaseous halogenated volatile anesthetics. 35 mutations in the ryanodine receptor gene RYR1 have been validated to trigger fulminant MH, and 260 RYR1 variants are suspected to account
for 60-70% of MH cases, making genetic testing impractical. Currently, the only clinically accepted diagnostic test for MH
worldwide is to submit an invasive muscle biopsy to in vitro contracture testing (IVCT) using halothane, or caffeine. The
goal of our study is to develop a less invasive rapid throughput assay based on primary myotubes cultured from small punch
biopsy to quantitatively determine sensitivity to challenge with caffeine and halothane or isoflurane singly or in combination. Our laboratory has produced transgenic mice expressing known human MH mutations within the N-terminal (R163C),
middle (R2435H), and C-terminal (T4826I) regions of the RyR1 sequence. Myoblasts have been isolated from wildtype
mice, and mice that are either heterozygous (HET) or homozygous (HOM) for the MH mutations. They were expanded in
the presence of FBS and bFGF and seeded onto 96-well plates. Cells were deprived of growth factors to promote differentiation, and were loaded with the Ca2+ indicator Fluo-4. Sarcoplasmic Ca2+ concentration was imaged in real-time using
two methods: (1) Slow-throughput epifluorescence microscopy of individual muscle cells, (2) Rapid throughput imaging of
muscle cell populations measured using FLIPR TETRA platform. We expect that cells from mutant mice show enhanced
responses to triggering agents permitting reliable preclinical testing.
Research Grant: Supported by the National Institutes of Health (NIH) grant 1P01 AR52354 to I.N. Pessah
Student Support: Supported by a VSTP fellowship from Merial
283
In vivo evaluation of a novel HER2 inhibitor, lapatinib, in osteosarcoma
Kim T. Tran, Tasha Miller, Heather M. Wilson-Robles
Veterinary Medical Scientist Research Training Program (Tran) and Department of Small Animal Clinical Sciences
(Miller, Wilson-Robles), College of Veterinary Medicine and Biomedical Sciences, Texas A&M University,
College Station, TX
Osteosarcoma (OS) is the most common primary bone malignancy in both humans and pet dogs. This disease accounts
for 8% of canine tumors and for over 400 new diagnoses in children in the U.S. each year. Approximately 15-20% of OS
patients (human and canine) have metastatic disease at diagnosisthough over 80% will go on to metastasize eventually.
Current multimodality therapy consists of radical surgery and systemic chemotherapy. Even with aggressive therapy the
median survival time for dogs is only 10-12 months and in humans the 5-year survival rates for patients with metastatic or
recurrent disease remain poor. Lapatinib is a small molecule inhibitor of human epidermal growth factor receptor 2 (HER2),
which is over-expressed in the cytoplasm of approximately 85% of canine and 62% of human OS. Lapatinib, unlike trastuzumab, works intracellularly to inhibit this receptor. Inhibition of HER2 is associated with decreased tumor proliferation
and increased tumor cell death in both canine and human OS cell lines in vitro. Using subcutaneous xenografts of canine
and human highly metastatic OSA cell lines, Abrams and 143B, in athymic nude mice, comparisons of tumor size, micrometastasis, and HER2-related gene expression were made between control and treatment mice. Briefly, the data suggest that
lapatinib significantly reduces HER2 expression, the incidence of lung metastasis, and the quantity of canine DNA in murine
lungs (an expression of tumor burden). Thus far, however, the data indicate that lapatinib does not affect primary tumor size
or AKT1 gene expression.The results of this study will enhance the understanding of the mechanism of lapatinib as a novel
therapy for OS.
Research Grant: Fred and Vola Palmer Chair in Comparative Oncology
Prevalence of Failure of Passive Transfer, Dehydration, and Health Outcomes in Veal calves on Day of Arrival
Deanna Trearchis, Jessica Pempek, Margaret Masterson, Gregory Habing, and Katy Proudfoot
Department of Veterinary Preventative Medicine, College of Veterinary Medicine, The Ohio State University,
Columbus, OH
Young veal calves are at a high risk for disease and morbidity in early life. By identifying calves with failure of passive
transfer (FPT), dehydration, and other health concerns, we can potentially reduce the prevalence of early mortality. Our aim
was to determine the prevalence of FPT, dehydration, and clinical health outcomes in veal calves on the day of arrival to veal
farms. Health exams were conducted on thirty four calves from 6 veal farms (n = 204); exams included a blood sample to
determine packed cell volume (PCV) and total protein (TP) to evaluate dehydration and FPT, respectively. In addition, rectal
body temperature, signs of diarrhea (using a 0 to 3 scale), navel inflammation (0 to 3 scale), and skin tenting (present if the
tent remains for $ 4 sec) were recorded. Descriptive statistics were used to determine the proportion of calves with FPT
(TP < 5.5 g/dl), dehydration (PCV > 46% or a present skin tent), clinical diarrhea ($ 2 with and without a rectal temperature
$ 103F), and navel inflammation ($ 2). No calves reached our criteria for dehydration (PCV mean = 32, min = 7, max =
46); however, 22% were considered dehydrated when using a skin tent. Upon arrival, 5.4% of calves had FPT (TP mean =
6.5, min = 5.0, max = 8.8). Only 2.0% of calves presented with diarrhea, and of these, half had elevated body temperatures.
A quarter (25%) of the calves had inflamed navels, and 5% also had a high rectal temperature. Although we expected more
cases of dehydration and FPT, the high prevalence of navel inflammation is a novel finding from the study. Further research
is needed to determine the cause of navel inflammation, and the impact of poor health on the day of arrival on morbidity
and mortality later in life.
Research Grant: Research Grant: National Institute of Food & Agriculture 2015-36100-06083 (Proudfoot, PI)
284
A Novel Mechanism that Regulates the Surface Levels of NKp46 on Natural Killer Cells
Mete Ender Tuncay, Hemant Mishra, Connor Ulrich, Adam Mielke, Bruce Walcheck
University of Minnesota Department of Veterinary and Biomedical Sciences, St. Paul, MN
Natural Killer (NK) cells are cytotoxic lymphocytes of the innate immune system that target tumors and virally infected
cells. Unlike other lymphocytes, such as T cells, NK cells are able to kill target cells without prior sensitization. NK cell
stimulation occurs when the balance of signals from activating and inhibitory receptors on their surface is tipped towards
activation. NKp46, also referred to as CD335, is a cell receptor expressed at high levels on NK cells, which in humans is
encoded by the natural cytotoxicity triggering receptor 1 (NCR1) gene. NKp46 is one of several receptors that play a key
role in activating NK cells. It is known that the cell surface levels of NKp46 can be down-regulated by a metalloprotease
upon NK cell activation. The objective of this project is to better understand the underlying mechanism responsible for
NKp46 down-regulation. In our initial activation assays, we found that the nonspecific activating agent PMA induced the
down-regulation of NKp46 from the surface of human NK cells, but not the expression of NK2GD, another important
activating receptor expressed by these cells. When NK cells were initially treated with a selective ADAM17 inhibitor, this
blocked the down-regulation of NKp46 upon NK cell activation. Use of an ADAM10 inhibitor had no effect on NKp46
regulation. Our findings thus far suggest that ADAM17, but not ADAM10, has a critical role in down-regulating NKp46 by
a cleavage process. We are currently looking at different physiologically relevant stimuli of NK cells, as well as NKp46 on
canine NK cells. By understanding the underlying mechanism by which NKp46 is down-regulated, it may be possible to
block this process to enhance NK cell killing.
Research Grant: Research Grant: National Institutes of Health Grant, Award Number R01CA203348
Student Support: Student Support: National Institutes of Health, Award Number T35OD011118
Characterization of the fecal microbiome from EHEC positive and digital dermatitis negative beef cattle
Tiffani Turinski, Megan Kulow, Ermias Amene, Kelly Anklam, and Dorte Dopfer
School of Veterinary Medicine, University of Wisconsin, 2015 Linden Drive, Madison, WI
Digital dermatitis (DD) causes ulcerative lesions on the feet of cattle that are painful and lead to economic losses. Additionally, cattle can carry and introduce the zoonotic foodborne pathogen enterohemorrhagic Escherichia coli serotype O157:H7
(EHEC O157) into the human food supply. EHEC O157 does not result in clinical disease in cattle, but can cause human
illness ranging from diarrhea to potentially life-threatening hemolytic uremic syndrome. The mechanisms behind a cow’s
ability to shed EHEC O157 are not well-understood, but the ease of transmission of this disease is impacted by factors that
change when cattle are lame from DD infection, such as their contact structure. Preliminary data has shown that EHEC
O157 positive and DD negative infection status in beef cattle is associated with differences in their gut microbiome. We
hypothesize that obtaining a better understanding of fecal microbiome characteristics in EHEC+/DD- beef cattle will allow
us to quantify associations between EHEC+/DD- infection status and production variables. Fecal material from EHEC+/
DD- was collected from two nutritional supplement field intervention trials. DNA was isolated from these fecal samples
and sequenced. Raw sequence data was quality checked and filtered using mothur, a statistical software. Cleaned sequences
were aligned to the SILVA reference database to produce OTU classification and taxonomic assignments. Mothur outputs
were then exported into R for further analysis. The findings from this study will provide a better understanding of the fecal
microbiome characterizing EHEC+/DD- cattle, which may yield knowledge about how to reduce EHEC O157 contamination in the human food supply and thus reduce human infections.
Research Grant: Food Research Institute, UW-Madison
Student Support: Walter and Martha Renk Endowed Laboratory for Food Safety
285
Fescue toxicosis in grazing beef cattle: impact of environmental temperature and humidity
Zachary B. Turner, Ryan S. Mote, Nicholas S. Hill, Zachary P. Sanders, Dean P. Jones, Nikolay M. Filipov
Department of Physiology and Pharmacology (Turner, Mote, Filipov), College of Veterinary Medicine, University of
Georgia, Athens, Georgia; Department of Crop and Soil Sciences (Hill, Sanders), College of Agriculture, University of
Georgia, Athens, Georgia; Department of Medicine (Jones), Emory University School of Medicine, Atlanta, Georgia
Epichloe coenophiala, a symbiotic endophyte, infects tall fescue (Lolium arundinaceum), the predominant grass forage
in the Southeast U.S., and provides multiple benefits, but also produces toxic ergot alkaloids (TEA) that are causative for
fescue toxicosis in grazing herbivores. TEAs are monoaminergic agents and, upon ingestion, cause metabolic dysregulation and thermoregulatory impairment. In grazing beef this leads to decreased weight gain and marked economic losses,
especially when fescue grazing is during high environmental temperatures (T°) and humidity conditions. The nature of the
TEA- T° interaction under field conditions is unknown, so in the current study we evaluated the effects of grazing toxic
fescue under no heat stress (low T°-humidity index [low THI]) and under low/moderate heat stress (high THI). Angus steers
(n=12; 6 paddocks; 2 steers/paddock) were randomly assigned to toxic (Tox; 3 paddocks) or non-toxic (NT; 3 paddocks)
treatments. Urine, blood, feces, respiration rates (RR), surface, skin, and rectal T° (RT) were collected before, 1, 2, 11 (low
THI), 15 (high THI), 19 (high THI), and 25 (low THI) days after pasture placement; the weight of the steers was recorded
before and at the end of the study. From data analyzed so far, TEA consumption decreased weight gains and increased both
RR and RT independent of THI. Surface and skin T° were both significantly increased by high THI regardless of fescue
treatment, except for the ear temperature, which was increased the most in Tox steers under high THI (surface) or under both
low and high THI (skin). These findings indicate that THI’s effect on some thermoregulatory components is exacerbated by
TEA, while TEA’s effects on RR and RT are THI-independent.
Research Grant: USDA, National Institute of Food and Agriculture, Grant Number 67030-25004
Carnoy’s vs. Formalin: a comparison of two fixatives in colonic samples from weaned piglets
Clara Bush Vadala, Paula Giaretta, Anna Blick, Sarah Sprayberry, Chad Paulk, Todd Callaway, Jason Gill,
and Raquel Rech
Department of Veterinary Pathobiology, College of Veterinary Medicine and Biomedical Science (CB, PG, AB, RR)
and Department of Animal Sciences (CP, SS, JG), Texas A&M University , U.S. Department of Agriculture (TC),
College Station, TX.
The type of fixative used can impact the preservation of the intestinal mucosa, thickness of the mucous layer, and spatial distribution of the bacterial populations in histologic sections. In this study, we fixed colonic samples from twelve 4-week-old
piglets in Carnoy’s solution for 45 minutes and in 4% formaldehyde for 24 hours. After processing, slides were stained with
Hematoxylin and Eosin (H&E) and Alcian Blue (AB) stains. For intestinal morphometry, we performed 13 measurements
of the mucosal height in each H&E slide. We measured the thickness of the inner colonic mucous layer in 15 areas and
counted the number of goblet cells in 15 crypts for each AB slide. Statistical analysis was performed using one-way ANOVA
test, with P<0.05 considered statistically significant. Comparing intestinal morphometry, the mucosal height in Carnoy’s
fixative was significantly increased (437.16 6 147.77 mm, P<0.05) compared with 4% formaldehyde (405.68 6 117.04 mm,
P<0.05). The number of goblet cells in Carnoy’s fixative (37.01 6 12.52, P>0.05) and 4% formaldehyde (34.75 6 12.70,
P>0.05) did not show significant differences. In contrast, the thickness of the inner mucous layer was significantly greater
in Carnoy’s fixative (3.11 6 1.01 mm, P<0.05) compared with 4% formaldehyde (1.48 6 1.01 mm, P<0.05). In conclusion,
Carnoy’s fixative better preserves the mucous layer while slightly expanding the colonic mucosa. Future studies will compare the number of bacteria on the mucosal surface via FISH using both fixatives. Ultimately, the use of an optimal fixative
will be critical for defining the significant and biologically meaningful spatial relationships of the microbiota in the gut.
Research Grant: Bill Gates Foundation
Student Support: Texas A&M One Health On-Campus Summer Research Program
286
Survey of infectious diseases in free-ranging Chilean Humboldt penguins (Spheniscus humboldti)
Ashley van Batavia, Samuel D. Sibley, Roberta E. Wallace, and Tony L. Goldberg
Department of Pathobiological Sciences, School of Veterinary Medicine, University of Wisconsin-Madison, Madison, WI
(van Batavia, Sibley, & Goldberg); Milwaukee County Zoo, Milwaukee, WI (Wallace)
The International Union for Conservation of Nature (IUCN) classifies Humboldt penguins as a vulnerable species at risk of
extinction, with reports of population decline. The emergence of novel pathogens has been shown to impact the health of
other penguin populations, yet few studies to date have examined health-related threats to Humboldt penguins, and none of
these have applied advanced genomics methods. Census data and blood samples collected from free-ranging Humboldt Penguins (Spheniscus humboldti) in Chile from 1994-2003 were examined via next generation (“deep”) sequencing techniques
to investigate the potential for viral causes of population decline. This study expands the breadth of viral detection beyond
serologic testing using “unbiased” methods capable of detecting all DNA and RNA viruses in a sample. This contrasts with
traditional gene-specific PCR methods, whereby DNA or RNA targets are chosen a priori. Using these techniques, we expect to identify novel viruses that have not yet been described in penguins, identifying potential disease risks and establishing baseline health parameters of the Humboldt penguin population and future diagnostic targets that will aid conservation.
Research Grant: Milwaukee County Zoo.
Development of a Canine Functional Questionnaire for Assessing Rehabilitation Outcomes
Marjan van den Hoogen, Jessica Rychel, Wendy Herlihy, and Kevin K. Haussler
Utrecht University, Utrecht, The Netherlands (van den Hoogen), Fort Collins Emergency and Rehabilitation Hospital,
Fort Collins, CO (Rychel, Herlihy) and Colorado State University, Fort Collins, CO (Haussler)
Physical therapy and rehabilitation for dogs is rapidly growing; however, there are limited methods to objectively evaluate
treatment effects. The presence of pain is only one small component of functional limitation in dogs. Additional measures
of proprioception, flexibility, motor control and strength are needed to fully assess a return to before-injury status. The
objective of this study was to develop a functional questionnaire for assessing changes in musculoskeletal and neurologic
status in response to physical therapy. Our hypothesis was that a questionnaire could reliably identify normal and abnormal
musculoskeletal and neurologic function, localize affected body regions, and quantify changes in functional status over time
when compared to a validated pain questionnaire (Canine Brief Pain Inventory, CBPI). Clients completed questionnaires on
functional capabilities of their dogs (N=95) during rehabilitation sessions. Dogs (aged 8.363.7 years, 44% females, 56%
males) also completed functional tasks specific to the axial skeleton, thoracic or pelvic limbs. Primary musculoskeletal
disease occurred in 53% (N=50) and neurologic disease was reported in 47% (N=45) dogs. Significant group differences in
overall function (t-test, p<0.05) were found for normal (060) versus abnormal (2065) function. No significant differences
were found between affected body regions (ANOVA). Significant pre- and post-treatment differences in CBPI and affected
body regions scores occurred (ANOVA). Our functional questionnaire may be able to capture specific changes in musculoskeletal or neurologic disability that are not readily detected by the CBPI as some rehabilitation patients may have minimal
pain, but significant loss of function.
Research Grant: Orthopaedic Research Center, Colorado State University
Student Support: Merial Veterinary Scholar International Program
287
Comparison of virus isolation from fabric for use in herd surveillance of viral bovine respiratory disease
Ashley N. Varley, Amelia R. Woolums
Dept. Of Pathobiology and Population Medicine (Varley, Woolums), Mississippi State University College of Veterinary
Medicine, Mississippi State, MS
Bovine respiratory disease (BRD) is the leading cause of morbidity and mortality in many classes of cattle, including
weaned dairy calves, feedlot cattle, and beef calves between 3 weeks of age and weaning. Both bacterial and viral agents
have been found to cause clinical signs. Because sampling individual calves in cow-calf operations is logistically difficult,
this study forms the basis for a possible method of herd surveillance for viral BRD in cow-calf operations. To ensure efficacy
in a pasture, the proposed method was first tested in a laboratory setting. Cotton and polyester fabrics were purposely inoculated with Bovine Herpes Virus- 1. Four drying times (0h, 4h, 12h, 24h) and two methods of bacterial control (antibiotics
in eluent and 0.22um filter) were compared. At each drying time, four test groups were evaluated; cotton with eluent treated
with antibiotics, cotton with filtered eluent, polyester with eluent treated with antibiotics, and polyester with filtered eluent.
Virus isolation on Madin Darby Bovine Kidney cells was used to for virus recovery. At time 0h, no significant difference
was found in virus isolated in each treatment group. At time 4h and 12h, no virus was obtained from polyester; virus was
isolated from cotton but there was no difference between either bacterial control method. At 24h, cotton with antibiotics as
a bacterial control method led to improved virus isolation as compared to the other 3 methods
Research Grant: Mississippi State University CVM, Department of Research and Graduate Studies
Student Support: Mississippi State University CVM, Department of Research and Graduate Studies
Commensal Enterobacteriaceae Drive Variable Salmonella Colonization Resistance of Mice from Different Vendors
Eric M. Velazquez and Andreas J. Baumler
Medical Microbiology and Immunology, University of California, Davis, CA
The gut microbiota contributes to intestinal health and can protect its host against diarrheal infections. However, specific
members of the microbial community that confer protective benefits are not fully described. We hypothesized that specific
microbiota differences between healthy individuals can be associated with more resistance to enteric pathogens. We first
tested if genetically similar strains of mice obtained from different commercial vendors exhibit different responses during
Salmonella infection. C57BL/6 mice from Harlan and Jackson were orally challenged with increasing doses of Salmonella.
At each given dose, pathogen loads were consistently higher in Jackson mice compared to Harlan mice. Next, we directly tested if the difference in Salmonella colonization was microbiota-dependent. Colonizing germ-free mice with fecal
transplants recapitulated the infective dose response associated with each donor. Microbiome sequence analysis identified
Proteobacteria in donor and recipient animals from Harlan, but not Jackson. We investigated the causal role of these commensal bacteria by transferring them into Jackson mice. After being colonized with Enterobacteriaceae isolates, Jackson
mice showed improved intestinal resistance against Salmonella. Importantly, these findings suggest that the natural levels of
Enterobacteriaceae among healthy individuals may determine susceptibility during a foodborne outbreak.
Research Grant: NIH 5R01AI096528-05 “Microbiota Outgrowth by Salmonella”
Student Support: NIH 2T35OD010956-16 “Students Training in Advanced Research”
288
Using human cell-specific data and correlation to cell proportion to predict pig neutrophil-specific genes
Gianna Vella, Martine Schroyen, Crystal Loving and Christopher Tuggle
College of Veterinary Medicine (Vella) and Department of Animal Sciences (Schroyen, Tuggle), Iowa State University,
and NADC (Loving), Ames, Iowa
Identification and profiling of cell-specific genes provides researchers and medical practitioners a more vivid portrait of
healthy and diseased states in individual tissues. Currently, cell-specific gene profiling has been conducted in human and
mouse cells under both resting and inflammatory models. Genes specific to leukocyte type during and after differentiation
have been identified in human peripheral blood samples. Evidence of conservation of lymphoid and myeloid signature genes
in mice and humans has been published. In the absence of pig specific data, it is the intentions of this study to validate the
use of a combination of human cell-specific data and correlation of gene expression to cell type proprotion in pig whole
blood, as a predictor and guide in the deconvolution of pig cell-specific gene profiles from complex mixtures of multiple
cell types, such as in whole blood. Fisher’s Exact tests demonstrate significant overlap of pig genes whose expression in pig
whole blood is correlated with neutrophil percentage with a published human cell-specific gene list. This significant enrichment of genes highly correlated with neutrophil percentage with those in a published list of human leukocyte-specific genes
validates the use of human cell-specific gene lists to predict pig cell specific genes. Quantitative-PCR analysis of selected
genes in isolated porcine neutrophils validated the predicted neutrophil specific expression when compared to peripheral
mononuclear cell preparations. These data verify that human cell-type specific expression combined with gene expression
correlation to cell type proportion in whole blood can be used to predict similar expression patterns in swine.
Research Grant: National Institute of Health 1R24-OD019813-01
Student Support: National Institute of Heath T35 training grant 4T35OD012199-14
Estriol protects female mice against influenza A virus by reducing pulmonary inflammation
Meghan Vermillion and Sabra Klein
Molecular and Comparative Pathobiology, Johns Hopkins School of Medicine, Baltimore, MD (Vermillion) and
Molecular Microbiology and Immunology, Johns Hopkins School of Public Health, Baltimore, MD (Vermillion, Klein)
Estriol (E3) is a pregnancy-specific estrogen in humans and mice that is produced by the placenta. E3 has potent immunomodulatory functions and can ameliorate diseases caused by inflammation, including multiple sclerosis (MS). Like MS,
pulmonary disease associated with influenza A virus (IAV) infection is predominantly caused by aberrant inflammation
and immunopathology, rather than virus replication. We hypothesized that exogenous E3 may protect female mice against
IAV-associated disease by reducing pulmonary inflammation and tissue damage. Adult gonadally-intact female C57BL/6
mice received a subcutaneous implant of either 5mg E3 or placebo. This dose of E3 increased circulating concentrations to
pregnancy levels, as measured by mass spectrometry. Mice were intranasally infected with a sublethal dose of mouse-adapted 2009 H1N1 IAV, and body mass was recorded daily. Pulmonary inflammation and immune cell populations were evaluated by histology and flow cytometry at 9 and 14 days post-infection (dpi). Following infection with IAV, E3-treated females
lost significantly less body mass compared with placebo-treated females, which was associated with significantly less pulmonary inflammation at 9dpi. Moreover, E3-treated females had an increased ratio of CD4:CD8+ T cells, and CD4+ T cells
were skewed toward Th2 and away from Th1 and Th17 responses at 9 dpi. These data suggest that pregnancy levels of E3
are protective against IAV infection and indicate that the estrogen-driven immunomodulation during pregnancy is not the
cause of pregnancy-associated IAV disease severity. These data also provide evidence that E3 has broad immunomodulatory
effects and may be beneficial in the context of inflammation-driven disease.
Research Grant: NIH R21 AI112838 and Centers for Excellence for Influenza Research and Surveillance
HHSN272201400007C
Student Support: NIH T32 Training Grant OD011089
289
Rodent Models of Ozone-Induced Eosinophilic Rhinitis: Species- and Strain-Dependent Variations
Nicholas Vetter, Daven Jackson-Humbles, Ryan Lewandowski, James Wagner and Jack Harkema
Department of Pathobiology and Diagnostic Investigation, College of Veterinary Medicine, Michigan State University,
East Lansing MI
Epidemiological studies suggest that elevated outdoor concentrations of ozone, the most common air pollutant in photochemical smog, are associated with activation of eosinophils in the airways of children. Our laboratory has recently reported that mice repeatedly exposed to ozone develop nasal eosinophilic inflammation, mucous cell metaplasia, and type
2 immunity that are dependent on group 2 innate lymphoid cells, and not T or B lymphoid cells (i.e., non-allergic type 2
rhinitis). The present study was designed to further elucidate species- and strain-dependent differences (genetic variability)
in ozone-induced rhinitis. C57BL/6 and BALB/c male mice and F344 male rats were exposed to 0 ppm (filtered air) or 0.8
ppm ozone for 9 consecutive weekdays (4 h/day). One day after the end of the exposures, animals were sacrificed and nasal
tissues were processed for histopathologic examination and morphometric analysis. Ozone exposure caused eosinophilic
rhinitis with mucous cell metaplasia in both rats and mice. No nasal histopathology was found in filtered air-exposed controls. Ozone-exposed mice had greater (7-12x) eosinophil densities in the nasal mucosa as compared to similarly exposed
rats. Ozone-exposed rats had greater (5-7x) volume densities of mucosubstances in nasal epithelium as compared to similarly exposed mice. C57BL/6 mice had 2x greater amounts of mucosal eosinophils as compared to BALB/c mice. Magnitude
of ozone-induced mucous cell metaplasia in nasal epithelium was similar between the mouse strains. These findings in different species and strains of rodents suggest that activation of eosinophilic inflammation in the airways of children exposed
to ozone might depend on genetic as well as environmental factors.
Research Grant: Research Grants: ACC 6148 and EPA RD83479701
Student Support: Student Support: NIH Grant No. T35OD016477 to Michigan State University
Association of progestogens with inflammation and immunity in critically ill foals
Stephanie M. Vijan, Katarzyna A. Dembek, Stephen M. Reed, Ramiro E. Toribio
Department of Clinical Sciences, College of Veterinary Medicine, The Ohio State University, Columbus, OH
Sepsis is the leading causes of mortality in foals. Progesterone is mainly known for its role in pregnancy. However, it is
also a precursor to adrenocortical steroids and likely plays important functions in foals. Human studies demonstrated progesterone predisposes to inflammatory conditions. However, the mechanisms by which progesterone influences outcomes
in sick foals remains unclear. The goal of our study was to measure cytokine and serum amyloid A (SAA) concentrations in
foals and to determine their association with progesterone, severity of disease, and mortality. We hypothesized septic foals
will have higher progesterone, 17a-OH-progesterone and cortisol concentrations compared to healthy foals. We proposed
progesterone concentrations will be associated with an inflammatory response, disease severity and mortality. Foals (n=53)
were divided into 3 groups based on disease severity (septic, sick non-septic [SNS], and healthy) and 2 groups based on
survival. Blood samples were collected on admission and hormones were measured by radioimmunoassay. Progeste

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