TECHNICAL BULLETIN OF THE FLORIDA MOSQUITO CONTROL

Transcription

TECHNICAL BULLETIN
OF THE
FLORIDA MOSQUITO
CONTROL
ASSOCIATION
VOLUME 10, 2016
TECHNICAL BULLETIN
OF THE
FLORIDA MOSQUITO
CONTROL
ASSOCIATION
VOLUME 10, 2016
FLORIDA MOSQUITO CONTROL ASSOCIATION, INC.
ORGANIZED IN 1922
The Florida Mosquito Control Association, Inc. is a non-profit, technical, scientific, and educational
association of mosquito control, medical, public health, and military biologists, entomologists, engineers, and
lay persons who are interested in the biology and control of mosquitoes or other· arthropods of public health
importance.
TECHNICAL BULLETIN OF THE FLORIDA MOSQUITO CONTROL ASSOCIATION
EDITOR-IN-CHIEF: James E. Cilek, Ph.D.
E-mail: [email protected]
ASSISTANT EDITOR: Jonathan F. Day, Ph.D.
E-mail: [email protected]
ASSISTANT EDITOR: Nathan D. Burkett-Cadena, Ph.D.
Email: [email protected]
FMCA MEMBERSHIP
Individual membership fees for the Florida Mosquito Control Association (FMCA) are $35.00 per year and
student memberships are $15.00 per year, payable January 1 of each year. Life member, sustaining industry, and
sustaining governmental memberships are also available. For more information please visit the FMCA website:
floridamosquito.org and click on the tab “membership “or e-mail the Executive Director at: ExecutiveDirector@
floridamosquito.org
CORRESPONDENCE
Communications relating to membership, change of address, and other Association matters should be
sent to the Executive Director at: [email protected]. Communications relating to suggested
content of future volumes of the Technical Bulletin should be addressed to the Editor-In-Chief.
The Technical Bulletin of the Florida Mosquito Control Association is published by the Florida Mosquito
Control Association, Inc.
Printed by the
E. O. Painter Printing Company
P.O. Box 877
DeLeon Springs. FL 32130
ARBOVIRUS SURVEILLANCE
AND MOSQUITO CONTROL WORKSHOP
A volume of selected papers from:
The 11th workshop, March 25-27, 2014, and the 12th workshop, March
24-26, 2015, and Anastasia Mosquito Control District and its Collaborating
Organizations
Edited by:
Rui-De Xue
Sponsored by:
Anastasia Mosquito Control District
St. Johns County, St. Augustine, Florida
and
USDA-ARS, Center for Medical, Agricultural, and Veterinary Entomology,
Gainesville, Florida
TABLE OF CONTENTS
Introduction
Rui-De Xue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1
Review Paper
Host-seeking and blood-feeding behavior of Aedes albopictus
Rui-De Xue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 2
Research Articles
Arbovirus surveillance report in St. Johns County, Florida, 2008-2014
James H. R. Weaver, Catherine Lippi, Mohamed F. Sallam, Marcia K. Gaines,
and Rui-De Xue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 14
Spatial analysis of arbovirus transmission in St. Johns County, Florida
Mohamed F. Sallam, Catherine Lippi, and Rui-De Xue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . 18
Relationship between citizen knowledge, vegetation coverage, and frequency of
requests for mosquito control service in St. Johns County, Florida
Claudia A. Davidson, Jodi M. Scott, Tahjim H. Hossain, John C. Beier,
and Rui-De Xue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 30
Mosquito species composition and impact of trapping sites on floodwater mosquitoes,
Aedes vexans in Xinjiang, China
Mei-De Liu, Yan-De Dong, Gui-Lin Zhang, Zhong Zheng, Rui-De Xue,
and Tong-Yan Zhao. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 39
Lunar phase impact on Coquillettidia perturbans and Culex erraticus host seeking in
northern Florida
Yong-Xing Jiang. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 44
Field evaluation of three commercial mosquito traps and five attractants in
northeastern Florida
Ali Fulcher, Rachel Shirley, Michael L. Smith, Jodi M. Scott, and Rui-De Xue. . . . . . . . . . . 50
Field evaluation of Mosquiron 0.12CRD against Culex quinquefasciatus in storm
drains, downtown St. Augustine, Florida
Ali Fulcher, Rui-De Xue, Jodi M. Scott, Michael L. Smith, Marcia K. Gaines,
and James H.R. Weaver. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 54
Population reduction of mosquitoes and biting midges after deployment of mosquito
magnet traps at a large golf course adjacent to salt marsh habitats in St. Augustine,
Florida
Rui-De Xue, Whitney A. Qualls, and Dan L. Kline . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 59
Sublethal doses of attractive toxic sugar bait mixed with the insect growth regulator,
pyriproxifen did not effect survival or fecundity of Aedes albopictus
Codi Anderson, Jodi M. Scott, Ali Fulcher, Gunter C. Muller, and Rui-De Xue. . . . . . . . . . 64
Evaluation of power breezer and misting citronella against Aedes albopictus
Emily H. Thompson, Jodi M. Scott, Ali Fulcher, Michael L. Smith, Phil Koehler,
and Rui-De Xu. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 70
Bifenthrin barrier spray against Aedes albopictus around an urban cemetery,
St. Augustine, Florida
Christopher Bibbs, Codi Anderson, Michael L. Smith, and Rui-De Xue. . . . . . . . . . . . . . . . 76
Laboratory evaluation of seven insect repellents against the lone star tick,
Amblyomma americanum
Jodi M. Scott, Ali Fulcher, John M. Henlzer, and Rui-De Xue . . . . . . . . . . . . . . . . . . . . . . . . 81
Laboratory and field evaluation of OFF! CLIP-ON mosquito repellent device
containing metofluthrin against the lone star tick, Amblyomma americanum (Acari:
Ixodidae)
Rui-De Xue, Jodi M. Scott, Ali Fulcher, Whitney A. Qualls, John M. Henlzer, Marcia K.
Gaines, James H.R. Weaver, and Mustapha Debbou. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 85
Operational and Scientific Notes
Evaluation of Talent® UV light traps compared with CDC light traps with or
without dry ice to collect fresh and salt water mosquitoes in northeast Florida
Michael L. Smith, Whitney A. Qualls, and Rui-De Xue. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 91
Effects of leaf washing on the persistence of a sugar bait – pyriproxyfen mixture to
control larval Aedes albopictus
Jodi M. Scott, Ali Fulcher, Whitney A. Qualls, Gunter C. Muller, and Rui-De Xue. . . . . . . . 93
Field comparison of thermal fog application of sumithrin and barrier spraying of Talstar
against Aedes albopictus in residential yards, St. Augustine, Florida
Jennifer Gibson, Rui-De Xue, and Michael L. Smith . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 96
Workshop Programs
Program for the eleventh Arbovirus Surveillance and Mosquito Control Workshop,
AMCD, St. Augustine, Florida, March 25-27, 2014 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 99
Program for the twelfth Arbovirus Surveillance and Mosquito Control Workshop,
AMCD, St. Augustine, Florida, March 24-26, 2015 . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 105
INTRODUCTION
During 2014 and 2015 the Anastasia Mosquito Control District (AMCD) of St. Johns
County, Florida held its eleventh and twelfth
annual Arbovirus Surveillance and Mosquito
Control Workshops at District headquarters
in St. Augustine, Florida. Both workshops
were jointly sponsored by the AMCD and the
USDA, Center for Medical, Agricultural, and
Veterinary Entomology (CMAVE) in Gainesville, FL. These workshops were designed
to facilitate the exchange of information
regarding mosquito-borne diseases, review
recent research and developments in arbovirus and mosquito surveillance, mosquito
control, and to offer unique training opportunities for mosquito control professionals.
The eleventh workshop was held from
March 25-27, 2014, and included 66 presentations divided into 9 sessions. Due to the
outbreak of eastern equine encephalitis virus
(EEE) in the USA in 2013, the prevention and
control of this disease was the theme for this
workshop. Thomas Unnasch from University of South Florida, Tampa, was the keynote
speaker and presented some new insights into
the ecology of eastern equine encephalitis virus transmission in the southeastern United
States. Scientists from Mali, Malaysia, Saudi
Arabia, China, Brazil, and Israel, USDA/
CMAVE, Navy Entomology Center of Excellence, universities, as well as mosquito control
professionals from industry, state and local
governmental agencies provided presentations that included updates on mosquito surveillance and control and review of new mosquito control products and equipment.
The twelfth workshop was held from
March 24-26, 2015, and included 73 presentations divided into 7 sessions. Due to the
outbreak of chikungunya in the Caribbean
basin in 2014, the prevention and control
of this disease was the theme for this workshop. John Beier from University of Miami
Miller School of Medicine was the keynote
speaker and provided information on the
chikungunya outbreak and transmission
risk assessments. Randy Gaugler from Rutgers University, New Jersey was the domestic
guest speaker and presented a summary of
activities from the Center for Vector Biology
located on the main campus. Scott Ritchie,
from Australia, gave the international guest
speaker presentation on his work with honey
cards for arbovirus detection. Scientists from
USDA/CMAVE, Navy Entomology Center of
Excellence, universities, as well as mosquito
control professionals from industry, state and
local governmental agencies all provided presentations that included the latest updates on
mosquito surveillance and control methods
as well as control products and equipment.
A total of 40 continuing education credits (CEU) were provided to workshop attendees between 2014 and 2015. We are
especially appreciative of the speakers and
contributors who gave presentations in
these workshops and to those who submitted manuscripts for this volume, as well as
the organizations/companies who provided
partial funding for the workshops. We thank
those who reviewed manuscripts prior to
publication, including Lisa Drake*, Whitney
Qualls*, Mohamed Sallam*, Ali Fulcher*,
Jennifer Gibson*, Michael L. Smith, John
C. Beier*, Barry Tyler, Larry Hribar*, Peter
Jiang*, Gunter Muller*, Edith Revery, James
Cilek, Sandy Allan, Seth Britch, Muhammad
Farooq, Phil Kaufman, Tianyun (Steven) Su,
Andrew Li, Aaron Lloyd, Gregg Ross, Richard Weaver, Donald Barnard, Dan Kline, Jerry Hogsette, Jodi Scott*, Christopher Bibbs*,
and Charolette Hall* (* indicates reviewers
who reviewed more than one manuscript).
We also acknowledge the support and encouragement of the AMCD Board of Commissioners, administrative office, District
staff, and industry.
Rui-De Xue, Ph.D.
Director, Anastasia Mosquito Control
District
1
HOST SEEKING AND BLOOD FEEDING BEHAVIOR OF
AEDES ALBOPICTUS
RUI-DE XUE
500 Old Beach Road, St. Augustine, FL 32080
ABSTRACT. Aedes albopictus is a vector of
dengue and chikungunya viruses. In recent
years, this species has expanded its range to
the Americas, Europe, and Africa. Host seeking and blood feeding is one of the most important behaviors mosquitoes possess. Understanding both behaviors in Ae. albopictus
may result in more effective control of this
species from a nuisance standpoint, as well
as pathogen transmission. The topics discussed in this review include: olfaction, adult
ecdysis (as it relates to first host seeking attempt), frequency of host seeking and blood
feeding, biting cycle and blood feeding pattern, feeding location and biting behavior,
host preference, multiple blood feeding behavior, blood meal size, artificial blood feeding, and a few other miscellaneous factors
that have been reported to influence host
seeking and blood feeding behavior in this
species.
related to transmission of pathogens and has
driven the pathway of research to identify and
develop attractants and repellents for prevention and control of mosquitoes. This paper
reviews some of the progress made on Ae. albopictus behavior as it relates to finding and
successfully feeding on a suitable host.
II. OLFACTORY STRUCTURES
Visual, thermal, and olfactory stimuli all
contribute to host seeking and blood feeding behavior in female mosquitoes (Bowen
1991). The internal, external ultrastructure
(as well as possible biological features) of
virtually all olfactory sensilla on the antennae of Ae. aegypti (L.) and Anopheles stephensi
Liston have been summarized in a review
by Sutcliffe (1994). He found that small
and large sensilla coeloconica, ampullaceae, grooved pegs, and trichodea existed on
mosquito antennae, as well as capitate pegs
on mosquito palps. Sensilla coeloconica
and ampullaceae may respond to humidity and temperature while trichodea appear
to have a sensory function that may distinguish oviposition site-related compounds,
essential oils, or fatty acids associated with
human skin and certain repellents (Sutcliffe
1994). Grooved pegs were also reported to
have an olfactory function that responded
to ammonia, acetone, acetic acid, anisole,
and lactic acid. Capitate pegs on mosquito
palpi responded to carbon dioxide(Sutcliffe
1994). Subsequently, the type, distribution,
and fine structure of sensillae on the surface
of the antenna of Ae. albopictus was studied
using scanning and transmission electron
microscope methods by Huang et al. (1991),
Xue et al. (1991), and Seenivasagan et al.
(2009). One of the olfactory structures on
the antenna of Asian tiger mosquitoes was
Key Words: host seeking, blood feeding,
blood meal, Aedes albopictus
I. INTRODUCTION
The Asian tiger mosquito, Aedes albopictus
(Skuse), is an important vector of dengue
and chikungunya viruses and a competent
vector of several other arboviruses including
yellow fever, Ross River virus, West Nile virus,
La Crosse virus, and eastern equine encephalitis virus (Mitchell 1991, Ali & Nayar 1997,
Gratz 2004). A considerable body of work has
arisen since its invasion into the Americas and
Europe in the middle of the 1980’s (Hawley
1988, Francy et al. 1990, Rai 1991, Bonizzoni
et al. 2013). Host seeking and blood feeding in Ae. albopictus is a major part of female
mosquito behavior and a requirement for reproduction. This critical behavior is directly
2
Xue: Host Seeking and Blood Feeding Behavior of Aedes albopictus
sensilla trichoidea that possessed a pore at
their tip with many pores through the hair
wall. Other structures such as trichodea were
numerously distributed along all flagella segments of the antenna and consisted of long
or short hairs with sharp or blunt tips. In addition, long and short types of grooved peg
sensillae were also observed on the antenna.
Sensilla coeloconicae were observed in the
terminal flagellum of Ae. albopictus while
sensilla chaeticae were distributed throughout the body surface and revealed greater
variation in morphology and morphometric parameters (Seenivasagan et al. 2009).
The four types of sensillae from antenna of
Ae. albopictus are chactica (19%), trichodea
(71%), grooved pegs (9%), and coeloconica
(1%), based on their morphological characters, thickness of hair wall, pore density, and
dendrite number, respectively. Interestingly,
Huang et al. (1991) found that olfactory sensillae, in mosquitoes, may be stimulated by
host odors before any host seeking behavior
occurs.
III. ADULT ECDYSIS TO FIRST HOST
SEEKING
After adult emergence, juvenile hormone secretion plays an important role in
the physiological regulatory process in most
female mosquitoes before host seeking and
blood feeding (Huang and Xue 1991, Clements 1999, Meola and Readio 1988, Klowden
1990, Hansen et al. 2014). However, in Ae.
aegypti, host-seeking was either independent
of juvenile hormone concentration or the
sensitivity threshold was very different than
that of the ovaries (Bowen 1991) a process
that may be similar for Ae. albopictus.
Several workers have observed that the
time from adult Ae.albopictus emergence to
the beginning of host seeking and blood
feeding varied, depending on mosquito
strain. del Rosario (1963), Gubler and Bhattacharya (1971), Hien (1976), and Huang
and Gao (1991) reported that this period was
about 2-3 days, at 24-29° C, for strains from
Calcutta, Vietnam, Philippines, and Shanghai. The median time from emergence to
host seeking and blood feeding at 25° C was
3
2.5 days for an Ae. albopictus population from
Nagasaki, Japan, but this time increased to
3.5-4.5 days when adults were produced by
rearing larvae at high densities (Mori 1979).
IV. FREQUENCY OF HOST SEEKING
AND BLOOD FEEDING
Host seeking activity of Ae. albopictus,
as with any other species, is stimulated by
certain chemical odors and compounds
(Huang et al. 1991b, Hao et al. 2008, Guha
et al. 2014). Klowden and Briegel (1994) reported that host seeking was inhibited during the gonotrophic cycle (as measured with
an olfactometer). Furthermore, Fukumitsu
et al. (2012) reported that the concentration
of a neurotransmitter, dopamine, declined
in the head of Ae. albopictus as host-seeking
increased.
The behavioral sequence of host seeking
and blood feeding in Ae. albopictus is similar
to other species of mosquitoes; these steps
include, orientation, landing, probing, and
feeding as described by Hocking (1971), Edman and Spielman (1988), Bowen (1991),
and Takken (1991). Also, the frequency of
host seeking and blood feeding depends
largely upon longevity and number of gonotrophic cycles. Generally, female Ae. albopictus live from 4 to 8 weeks in the laboratory
at 25° C but may survive up to 3-6 months
outdoors (Hawley 1988, Xue et al. 2010). A
female can take about 1 week to complete
a gonotrophic cycle (the interval between
successive oviposition) (Hawley 1988) and
may complete 4 to 8 gonotrophic cycles during her lifetime, according to a laboratory
study by Fu et al. (1982). In order to complete these numbers of gonotrophic cycles,
Ae. albopictus females would need to seek
out hosts and feed at least 4 to 8 times (Fu
et al. 1982). However, this may not be absolute, because under laboratory conditions, a
Shenzhen strain of Ae. albopictus fed on a human subject twice during one gonotrophic
cycle and 10 times during 7 gonotrophic cycles (Liu Yang unpubl data). Field data from
Liu (1965) showed that the females, in their
study, had usually completed 1-2 gonotrophic cycles in their lifetime, according to the
4
Technical Bulletin of the Florida Mosquito Control Association, Vol. 10, 2016
number of observable follicular dilations.
But interpretation of field data must be approached with caution as the number of follicular dilations is often fewer than the actual number of gonotrophic cycles completed.
For example, no more than 3 dilations were
observed from field collected Ae. albopictus
even though those individuals were known
to have completed 4 or 5 gonotrophic cycles
(Liu, 1965). Moreover, if longevity of this
mosquito species is based on parity status
(i.e. gonotrophic cycles) the data may underestimate age (Mori and Wada 1977).
Time between oviposition events can
vary with geographic strains of Ae. albopictus.
Gubler and Bhattacharya (1971) reported
that a Vietnam strain of this mosquito species oviposited every 4.6 days at 26° C and
from 3-5 days at 25° C for a Shanghai and
Guangxi strain from China (Li, 1991). These
latter authors also noted that oviposition interval time was protracted at 20º C to at least
10 days and considerably shortened to 4.5-6
days at 30° C.
The numbers of blood meals, and frequency of host seeking, were negatively correlated with body size in Ae. albopictus (Hawley 1988, Farjana and Tuno 2013). Moreover,
several authors subsequently observed that
frequency of host seeking and blood feeding,
during a gonotrophic cycle, depended on a
variety of factors that included, mosquito
age (Xue et al. 1995), body size (Xue & Barnard 1996, 2012, Farjana and Tuno 2013),
blood meal size and mating status (Klowden,
1988, Lee and Klowden 1999, Barnard and
Xue 2009), parity and time of day (Xue and
Barnard 1996), carbohydrate availability and
fatigue (Xue and Barnard 1999, Xue and
Debboun 2014), host availability, species of
host and protective behavior, spatial repellents (Hao et al. 2008, 2013), and other miscellaneous factors (Xue et al. 1995, Xue and
Debboun 2014).
V. BITING CYCLE AND BLOOD
FEEDING PATTERN
Kawada and Takagi (2004) used a photoelectric sensor to record the nocturnal behavior of Ae. albopictus and found that host
seeking was positively correlated with increasing light intensity where the threshold intensity was >10 lux (approximately 1 foot candle)
(Kawada et al. 2005). However, these authors
added that complete darkness during the daytime deactivated host seeking behavior. Earlier, Hawley (1988) stated that Asian tiger mosquitoes primarily sought hosts and blood fed
during the day and rarely at night. Costanaza
et al. (2015) also found that host seeking activity varied with light levels (0 to 440 lux).
Consistent biting activity in their laboratory
studies occurred at a photoperiod of 12L:12D.
Kawada and Takagi (2004) further observed
that there were two peaks of diel activity in female Ae. albopictus one at 1000 to 1200 and
another from 1400 to 1600 h. Under complete
darkness, biting activity did not peak, but under a constant light regime, biting peaked at
2200-2300 (Huang and Gao 1991a, Xue 1991).
The USDA Gainesville, Florida strain, reared
under different photoperiod regimes in the
laboratory, did not show temporal significant
differences in blood feeding frequency. Also
short photoperiods did not affect blood-feeding activity of this strain (Table 1). Fu (1990)
also reported that outdoor biting activity
peaked in early morning (7-9 am) and late
afternoon (5-8 pm) in strains from Shanghai,
Table 1. Effects of photoperiod (at 25° C) on Aedes albopictus first gonotrophic feeding success using baby chickens
under laboratory conditions (Xue unpubl data)1.
Photoperiod regime (h)2
L:D=LL
L:D=14:10
L:D=12:12
L:D=08:16
L:D=DD
No. tested
Blood fed (% + SD)
195
400
240
240
240
74.9 + 6.2
66.8 + 2.0
64.2 + 4.0
59.2 + 4.0
65.5 + 7.6
Sugar water (3%) was provided when rearing adults. Six day old mosquitoes used in study, 1 hour blood feeding time.
Photoperiod regime of fourth instar larvae in rearing chamber: L=light, D=dark, LL=continuous light, DD=continuous darkness.
1
2
Fujian, Jiangsu, Guangxi and Henan, China. A
study by Delatte et al. (2010) also found that
Ae. albopictus exhibited a bimodal daily feeding activity in La Reunion where 89% of the
mosquito population was exophagic and 87%
were exophilic.
Peak biting activity of Ae. albopictus also appears to vary with location and habitat. A clear
bimodal pattern (early morning and late afternoon) of feeding activity has been observed
in Asian countries, such as China, Japan,
Philippines, and Singapore (Ho et al. 1973).
In southern Brazil, biting activity occurred
during early morning, early evening, and late
night with peaks at 6 am and 10 pm but the
greatest activity was between 4-5pm (Margues
& Gomes 1997). Interestingly, sugar feeding
and host seeking rhythm in Ae. albopictus, recorded in the laboratory using remote sensors,
discovered that the evening sugar feeding period was similar to the evening host seeking
period (Yee and Foster 1992).
In the laboratory, an indoor population
showed only a single broad period of activity in the afternoon (Wang, 1962). However,
the biting activities of a laboratory Shanghai
strain of Ae. albopictus showed that females
would feed on a host anytime during 24
hours regardless of long or short photoperiods (Huang & Gao 1991a). Also, the USDA
Gainesville laboratory strain of this species
will also seek, bite, and feed on human hosts
anytime during a 24 hour period but has early morning and late afternoon biting peaks
(Xue and Barnard 1996, 1997).
In summary, laboratory and field studies
have shown, without a doubt, that most strains
of Ae. albopictus are bimodal daytime biters
with a small peak in the early morning and a
larger peak in the late afternoon. Those strains
that do not show this bimodality are probably
highly inbred laboratory colonies reared under artificial light regime conditions.
VI. FEEDING LOCATION AND BITING
BEHAVIOR
Female Ae. albopictus prefer to bite exposed human skin primarily around the
ankles and knees (Robertson and Hu 1935).
For chickens, this mosquito species will usu-
5
ally bite the head, feet, and other exposed
skin, according to this author’s observations
(Xue unpubl). Although Ae. albopictus are
usually outdoor biting species, biting females can be collected from indoors.
Most female Ae. albopictus take 1-2 minutes to completed engorgement. Some
strains required 3.6 minutes to engorge
on a human arm (Soekiman et al. 1984 in
Hawley 1988). Fu (1990) found this species
took 20-30 seconds from probing to abdomen distention at 23-33°C and RH 68-95%.
The complete feeding period from probing
to engorgement averaged 104.14 seconds
(range 70-160 seconds). The temperature
and humidity change during experiments
did not significantly influence feeding persistence (Fu, 1990). Hien (1976) also found
that biting persistence of Ae. albopictus varied
according to strain, age, body size, host behavior, and environmental conditions.
VII. HOST PREFERENCE
Host preference in Ae. albopictus can directly affect vector competence and transmission risk of mosquito-borne disease
pathogens. Determination of host preference by mosquito vectors is usually obtained
by a variety of animal baits or traps (Takken
and Verhulst 2013). Blood meal source identification in Asian tiger mosquitoes can be
obtained by a variety of laboratory methods,
such as sera precipitin (Tempelis 1975) agar
gel-precipitin tests (Sivan et al. 2015), direct
or indirect enzyme-linked immunosorbent
assays (ELISA) using antisera (Ponlawat and
Harrington, 2005), and polymerase chain
reaction (PCR) (Richards et al 2006, Egizi et
al 2013).
In the laboratory, there are two methods
that can be used to determine host preference of female Ae. albopictus. Olfactometers
are often used to test host odors from different animals. Using this method, Miyagi
(1972) and Gubler (1970) determined that
Ae. albopictus preferred feeding on mammals, including mice/rats, guinea pigs, dogs,
cows, and humans. These authors also found
that chickens, snakes, turtles, and frogs can
be fed upon by this mosquito species (Miyagi
6
1972, Gubler 1970). A second method employed by Hess et al. (1968) used forage ratios for determination of host preference obtained by precipitin tests. His results showed
a mammal forage ratio >10 times that of
birds, but female Ae. albopictus could feed
on birds when mammals were unavailable.
Konishi’s (1989) laboratory experimental
results showed that Ae. albopictus preferred
to feed on humans compared with canines
when exposed concurrently. However, the
authors acknowledge that feeding pattern
in the laboratory and field were influenced
primarily by host availability.
In Japan and rural Singapore (Kim et al.
2009, Sawabe et al. 2010, Kek et al. 2014) Ae.
albopictus reportedly fed on mammals. In the
USA, this species preferred mammals and
birds (Passeriformes, Columbiformes, and
Ciconiiformes) as well as rats, dogs, humans,
rabbits, deer, squirrels, opossums, raccoons,
bovine, (Savage et al. 1993, Niebylski et al.
1994, Faraji et al. 2014, Valerio et al. 2010,
2010a). Asian tiger mosquitoes in Hawaii,
also preferred to feed on mammals, including dogs, cows, humans, cats, mongeese,
pigs, and horses (Tempelis 1975, Tempelis et al. 1970). In Missouri and New Jersey
Savage et al. (1993) and Faraji et al. (2014),
respectively, found only a small percentage
of blood meals from birds in their field collections. Sullivan et al. (1971) reported that
Ae. albopictus commonly fed on a variety of
hosts including humans, buffalo, pigs, dogs,
and chickens on the island of Koh Samui,
Thailand with an even higher percentage
of human feeding detected by sandwich-B
enzyme-linked immunosorbent assays (Ponlawat & Harrington 2005). These authors
ranked the blood feeding pattern of Ae. albopictus (top 4 host species) as follows: (1)
humans, (2) bovine, (3) swine, and (4) cat,
rat, chicken (equal numbers detected). They
concluded that nonhuman hosts provided a
significant source of the blood meals for Ae.
albopictus in Thailand. However, precipitin
tests, ELISA, and PCR were used to investigate host-feeding patterns of 172 blood-fed
Asian tiger mosquitoes collected from Potosi, Missouri, during the summers of 19891990. In that study, no blood meals from
cats, horses, rats, or swine were detected in
those mosquitoes.
The majority of blood meals (83.2%)
from Ae. albopictus in rural and semi-urban
areas of Singapore were from humans with
a small percentage from shrews, swine, dogs,
cats, turtles, and multiple hosts in rural settings (Kek et al. 2014). However, in that same
study blood meals from Asian tiger mosquitoes in the urban areas were entirely from
humans. Faraji et al. (2014) reported that
Ae. albopictus from Mercer County, New Jersey fed on mammalian hosts with over 90%
of their blood meals derived from humans
(58%) and domestic pets (23% cats and 15%
dogs). This mosquito species fed on humans
significantly more in suburban than in urban areas and cat-derived blood meals were
greater in urban habitats while were no avian-derived blood meals were detected . In a
central North Carolina suburban landscape
study, 7% of blood meals from Ae. albopictus
were avian hosts while 83% were mammalian that included humans (24%), cats (21%),
and dogs (14%) (Richards et al. 2006).
Blood from Asian tiger mosquitoes was exclusively human (100%) in urban zones
from Spain and southern Thailand and 95%
from Cameroon (Munnoz et al. 2011, Ponlawat and Harrington 2005, Kamgang et al.
2012, respectively). The results show that the
Ae. albopictus populations in the New World
are opportunistic feeders that use a variety
of hosts from cold-blooded to warm-blooded
animals (Delatte et al. 2010) This fact has
the potential for this mosquito species to become involved in the transmission cycle of
indigenous arboviruses (Savage et al. 1993).
Conversely, feeding preference may also
limit the vector potential of Ae. albopictus for
some arboviruses but the high mammalian
affinity of this mosquito species suggests that
it can be an efficient vector of mammal-and
human-driven zoonoses such as dengue, La
Crosse, and chikungunya viruses.
VIII. MULTIPLE BLOOD-FEEDING
BEHAVIOR
Aedes albopictus egg development is initiated by the blood meal (Xue et al. 2009).
This developmental process is linked to
the type of host, frequency, and volume
of blood that a mosquito can obtain. It is
known that anti-mosquito behavior by a
host can adversely influence the amount of
blood that mosquitoes will have available
to mature eggs (Walker and Edman, 1985).
Other factors related to the mosquito herself (i.e. body size, age, and pathogen infection), may result in reduced fecundity or
failure of egg maturation (Klowden 1988,
Xue and Edman 1991). For example, female Ae. albopictus blood fed on guinea pig
and human hosts produced significantly
more eggs compared with other species. Fecundity in mosquitoes that took a double
blood meal (chicken and guinea pig), a triple blood meal (3 separate guinea pigs), or
mixed blood meals (chickens, guinea pigs,
and humans) produced significantly more
eggs than mosquitoes fed once or twice
on a single chicken. Also, triple-feeding or
mixed feeding (3 meals on 3 host types)
decreased the gonotrophic dissociation
frequency in those mosquitoes (Xue et al.
2008, 2009). Gonotrophic dissociation occurs when eggs fail to mature after a blood
meal has been obtained [Xue et al. 2009].
Boreham (1976) suggested that multiple blood-feeding in vector mosquitoes
increased the probability of pathogen transmission. Because Ae. albopictus will engage in
multiple blood-feeding its’ risk of pathogen
infection and transmission is greater than
species that do not possess this ability. Indeed, Shroyer (1986), Lu (1990), Mitchell
(1991), Rai (1991), and Gratz (2004) have
found that Asian strains of this mosquito spe-
7
cies possessed a high susceptibility to several
arboviruses, including dengue, in the laboratory. Also, Xue and Barnard (1996) found
that multiple feeding and host-seeking behaviors were negatively correlated with body
size. Small body size tended to have more
contact with a host due to low nutrition
(Xue and Barnard 1996).
In the laboratory, Ae. albopictus must
feed on blood at least once for egg maturation. About 12.5% of a strain from Vietnam
required two blood meals to successfully
oviposit their first batch of eggs. Gubler and
Bhattacharya (1971) found that females in
the later gonotrophic cycles required two
blood meals, but never exceeded 22% of
the population. Mori (1979) reported the
number of blood meals required was dependent on mosquito strain, nutrition, host
behavior, and environmental conditions.
Klowden and Briegel (1994) also observed
that if Ae. albopictus was allowed to bloodfeed to repletion, its host-seeking behavior was inhibited during that gonotrophic
cycle.
In laboratory studies, Fu (1982, 1990)
found that about 13-50% of engorged Ae.
albopictus were able to take a second blood
meal from a mouse or human arm (Table
2). Furthermore, when Fu (1990) dissected
ovaries of females that had taken a second
blood meal 60, 75, and 100 hours after their
first blood meal they had produced about 98
to 102 eggs/female while 84.4% of those individuals had already laid some eggs. In the
field, 5% gravid females were collected from
human legs in biting collections whereas
37.8% gravid females were collected from
Table 2. Mean percentage of secondary blood meals from a Aedes albopictus Guangzhou, China strain under laboratory conditions (from Fu 1990).
Time after initial blood meal (h)
12
24
36
48
60
72
Mice
Human arm
No. tested Secondary feeding (%)
No. tested Secondary feeding (%)
198
199
200
199
200
100
aNA=not available
10.0
9.5
8.5
6.0
17.5
4.0
87
94
NAa
66
NA
71
23.0
37.6
NA
50.0
NA
12.7
8
human hosts indoors (Fu, 1982). In that
field study, 8.3% of collections contained
gravid females that had obtained a second
blood meal during the 24 h observation period. In that same study, second blood meals
by Ae. albopictus were observed in gravid individuals at each hourly sample interval during biting cycles.
Other field studies where multiple blood
feeding by Ae. albopictus has been reported
involve mixed blood meals from avian and
non-avian hosts (Richards et al. 2006) or
humans and other mammals (Delatte et
al. 2010). Ponlawat and Harrington (2005)
found that 3.8% of Ae. albopictus populations
contained a mixture of swine-human blood
while <1% of blood meals were dog-human
or cat-human. Multiple host blood meals
have also been reported from this mosquito
species collected from rural settings in Singapore (Kek et al. 2014). Also, Tempelis et
al. (1970) reported 0.3% mixed blood meals
in an Ae. albopictus population from Hawaii.
Molecular analysis (cytochrome b) of blood
meals from individuals of this species collected from 2 South Carolina zoological parks
revealed that the mosquitoes readily fed on
the captive animals, as well as the local humans and wildlife, resulting in mixed blood
meals (Tuten et al. 2012). Multiple blood
feeding by Ae. albopictus was reported in every possible combination from the animals
and humans present on La Reunion during
the chikungunya epidemic (Delatte et al.
2010). Therefore, the frequency of multiple
blood-feeding in Ae. albopictus should be further studied in the laboratory and field settings.
Hawley (1988) discussed the phenomenon of multiple feeding in Ae. albopictus
as a result of his field discovery during gut
dissections of gravid host seeking females.
Since, then several investigators have dissected the ovaries of this species from their
human biting collections and found varying
proportions of gravid females. Defining the
relationship of parity and number of blood
meals is confounded by two factors. These
factors have been pointed out by Hawley
(1988) and are, 1.) some gravid Ae. albopictus develop eggs without blood feeding and
2.) some host seeking gravid individuals
may have fed more than once before coming to bite the collector. Fu’s result (1982)
from laboratory experiments and Li’s observations from field studies (1991) have provided further evidence that gravid Ae. albopictus can take a second blood meal. Hawley
(1988) noted that while female Asian tiger
mosquitoes can take a second blood meal, it
was not necessary for successful egg laying.
However, Hawley further commented that if
the female did not have enough energy reserves in the larval stage then a second blood
meal may be essential for egg development.
IX. BLOOD MEAL SIZE
Blood meal size is dependent upon
mosquito strain, body size, age, host species and defense behavior, and environmental conditions (Klowden 1988, Xue &
Edman 1991). The mean blood meal size
in a Malaysian strain of Ae. albopictus was
2.6 mg (range 1.5-4.2mg) (from Hawley
1988). In a Vietnam strain, blood meal size
was 0.2 mg to 2.5 mg (Hien 1976) and in
a Chinese strain 1.23-1.83 mg (Fu 1990).
Blood meal size from a Japanese strain was
1.2-1.6 ul on unrestrained dogs with 45.264.3% of the mosquitoes engorging (Konishi 1989, Konishi and Yamanishi 1984).
Blood meal size in Ae. albopictus also affects
host seeking activity. Barnard and Xue
(2009) state that there was a curvilinear relationship between blood meal volume in
partially fed Ae. albopictus and host avidity
with a threshold between 0.8 and 1.0 mg
that reduced host seeking.
X. ARTIFICIAL BLOOD FEEDING
Laboratory colonization of Ae. albopictus
is important and necessary for providing research material for a variety of R&D projects.
In addition, mass rearing of this species for
use in conventional sterile insect release or
RIDL (release of insects carrying a dominant
lethal) programs for population replacement require a different mass feeding system for adult to egg production. Currently,
colonization of any blood feeding mosquito
requires vertebrate blood to optimize egg
production. Wind tunnel studies have shown
that carbon dioxide, water vapor, warmth,
and adenosine triphosphate (ATP) as stimuli that induce host seeking and blood feeding activity (Klun et al. 2013). Generally, Ae.
albopictus is a difficult species to rear when
using artificial techniques (Lyski et al. 2011).
However, this has not deterred a variety of
workers from trying. Usually sausage casings,
or lambskin condoms containing warm bovine, or other animal, blood have been used
for artificially feeding several species of mosquitoes, including Ae. albopictus. Interestingly, there are instances this mosquito species
has preferred sausage casings over lambskin.
Ultrastructural analysis revealed that sausage
casings have a textured surface while lambskin membranes do not.
Parafilm membrane stretched and
pressed into fiberglass window screen to
form a packet for holding warmed blood to
feed Ae. albopictus colonies have also been
used successfully by Tseng (2003). Ooi et al
(2005) found that cattle skin was the most favorable membrane to use after comparative
tests with the skin of chicken, fish, and salt
sausage. A 32-46% increase in blood feeding
was observed when Ae. albopictus were fed using horizontal or vertical blood packets (Lyski et al. 2011). Also, use of bovine collagen
sausage membranes in a vertical feeding position will increase the number of engorged
females and may be an additional factor
influencing feeding success of Ae. albopictus
(Lyski et al. 2011). A novel system by Deng et
al. (2012) consisted of a collagen membrane
casing filled with pathogen-free guinea pig
blood warmed by a heating device that yielded the same fecundity, survival rate, and egg
hatchability when compared with live guinea
pigs. Luo (2014) also used this method and
found that Ae. albopictus fed on whole pig
blood with ATP achieved an engorgement
rate of 84% compared with 51% from live
mice.
Other studies have shown that blood
serum and bovine serum albumin, not hemoglobin, may replace vertebrate blood
in artificial diets for mass mosquito rearing (Gonzales et al. 2015). However, a re-
9
cent blood-free protein diet, using a membrane feeding system, was evaluated by Pitts
(2014). In that study, Ae. albopictus accepted
the diet and produced eggs in greater numbers than cohort females fed with whole human blood. This implied that a readily available blood free diet could be utilized in the
laboratory for rearing this species. Elimination of handling blood, reduced animal cost,
and consistency of diet should potentially be
advantageous to mosquito rearing and mass
rearing facilities (Pitts 2104).
XI. OTHER INFLUENCING FACTORS
Host seeking and blood feeding behavior in Ae. albopictus is a complex behavioral
process. Friend and Smith (1977) have previously reviewed several factors affecting
feeding by bloodsucking insects. Since that
time, several additional studies have been
conducted on this species and are as follows.
Host seeking and blood feeding in Ae. albopictus is influenced by human physiological
differences, host species, and host defensive
behavior. Shirai et al. (2004) reported that
human subjects with blood group O attracted more Asian tiger mosquitoes than other
blood groups (i.e. B, AB, and A) but was
only significantly more attractive than blood
group.
Xue (unpubl) found that host defensive
behavior significantly affected blood feeding
success in Ae. albopictus where the percentage of blood-feeding mosquitoes on a restrained host (chicken) was greater than an
unrestrained one in laboratory studies (Table 3). Also, Xue and Barnard (1996) found
that host attacking behavior of Asian tiger
mosquitoes was influenced by age and body
size. Large bodied, older females exhibited a
higher frequency of biting humans than did
younger, small body ones.
Temperature remains a main factor
influencing host seeking and blood feeding activity in Ae. albopictus. A Chinese
strain of this species will feed in a temperature range of 12-35° C in the laboratory. Fu (1990) found that feeding frequency was increased when temperature
increased, but a thermal threshold was
10
Table 3. Effect of temperature on mean percent blood-feeding by Aedes albopictus on restrained and unrestrained
chickens under laboratory conditions (Xue unpubl data).
Restrained chickens
Temperature (°C)
15
20
25
27
30
Unrestrained chickens
No. tested
Feeding (%)
No. tested
Feeding (%)
148
149
151
150
150
3.4 ± 1.2
35.7 ± 10.4
76.7 ± 3.1
69.3 ± 5.0
87.3 ± 5.0
300
280
250
250
150
0.7 ± 1.2
5.7 ± 2.1
1.7 ± 2.7
3.0 ± 0.6
7.3 ± 2.3
t=3.414, df=4, P<0.05 (between restrained and unrestrained hosts).
met at 35°C that decreased feeding. The
USDA, Gainesville, Florida strain of Ae. albopictus showed, in laboratory studies, that
blood-feeding increased as temperature
increased (Table 2). Similar host seeking
activity occurred in a Shanghai strain at
17-27° C (Huang and Gao 1991a). Conversely, a seasonal reduction in emergence
of host seeking females was observed at a
minimum threshold temperature of 13° C
in northern Italy (Roiz et al. 2010).
In conclusion, host seeking and blood
feeding in Ae. albopictus are two of the most
important, and exceedingly complex, behaviors that this, or any, mosquito species possess. Both behaviors are influenced by many
factors whether they are studied in the field
or laboratory. Understanding these processes
and identifying the weak links in these relationships, whether it is nuisance and/or disease transmission, will result in more effective
management of this species in the future.
XII. ACKNOWLEDGEMENTS
The author thanks C. Hall, C. Bibbs, L.
Drake, and J. Beier for editing and review
draft manuscripts, and the Editors of the
TBFMCA for their editorial review of the
manuscript.
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ARBOVIRUS SURVEILLANCE REPORT IN ST. JOHNS
COUNTY, FLORIDA, 2008-2014
JAMES H.R. WEAVER, CATHERINE LIPPI, MOHAMED F. SALLAM,
MARCIA K. GAINES, AND RUI-DE XUE
ABSTRACT. Arbovirus surveillance is a core
function of the Anastasia Mosquito Control
District in St. Johns County, Florida. This
agency largely relies on the use of sentinel
chicken flocks to monitor annual incidence
of arbovirus transmission. Environmental
and arbovirus surveillance data for the years
2008-2014 were analyzed. Total monthly
rainfall and mean monthly temperature
were found to have significant relationships
with eastern equine encephalitis seroconversions in sentinel chicken flocks, while only
temperature was found to have a significant
influence on West Nile virus seroconversion.
The identification of environmental and
temporal patterns that influence transmission of arboviruses is vital to the formation
of effective vector management programs
for the District
programs with the goals of alleviating the
risk of disease transmission and mitigating
disease burden. In the last decade, local concerns have increased over mosquito-borne
diseases since the first record of West Nile
virus (WNV) in 1999 in New York (Hayes
et al., 2005) and its eventual spread to St.
Augustine in 2001 (Connelly et al. 2012).
In addition to public health concerns, the
presence of mosquito-borne diseases can
have significant economic impact. St. Johns
County’s local economy is primarily driven
by tourism. However, in spite of the recent
national economic downturn, the county is
considered one of the fastest growing areas
in the United States (SJC TDC, 2015). In
light of this fact, AMCD continues to stress
the importance of their arbovirus surveillance program as a critical part of control
activities that provides economic and public
health benefits for the county.
Florida mosquito control arbovirus surveillance, as administered by most local programs,
continues to serve as an early warning system
for mosquito-borne viral diseases. This program is part of the “Florida Sentinel Chicken
Arbovirus Surveillance Network” established
in 1978 (O’Bryan and Jefferson, 1991). The
Surveillance Network is based on using sentinel chicken flocks to monitor epizootic virus
transmission by infected mosquitoes. Anastastia Mosquito Control District participates in
this program where data from chicken viral
seroconversion rates, together with mosquito
surveillance data, provide local public health
and mosquito control officials the needed information to assess the frequency and intensity
of epizootic virus transmission.
Moreover, consistent and appropriate
mosquito collection data, coupled with ar-
Key Words. West Nile virus, eastern equine
encephalitis virus, sentinel chicken, vector
mosquitoes
I. INTRODUCTION
St. Johns County lies in northeastern
Florida with a total area of 1,588 km2 (609 sq.
miles) and is located between the St. Johns
River to the west and Atlantic Ocean to the
east. As a result, there is great diversity in
salt and fresh water habitats throughout the
County that provides developmental habitats for approximately 42 mosquito species.
Eleven of these mosquito species are known
vectors of mosquito-borne pathogens. Within this jurisdiction, the Anastasia Mosquito
Control District (AMCD) oversees the entire
county’s mosquito surveillance and control
14
Weaver et al.: Arbovirus Surveillance Report in St. Johns County, Florida, 2008-2014
boviral surveillance, provides the necessary
justification for the application of vector
control measures for AMCD. Previously, Xue
and Qualls (2008) reported arbovirus surveillance results for St. Johns County from 2001
through 2007. In this report, we present the
results and analysis of AMCD’s mosquito arbovirus surveillance program for West Nile
(WNV), eastern equine encephalitis (EEEV),
and Highlands J (HJ) viral diseases in St.
Johns County from 2008 through 2014.
II. MATERIALS AND METHODS
Sentinel Chicken Surveillance. Sixty 21-week
old chickens were distributed over 10 locations in the county at the beginning of April
through December every year to monitor
arbovirus activity. Approximately 2.0 ml of
blood was taken from each chicken’s wing
vein once a week. Blood samples were kept
in labeled vacutainers (Fisher Scientific) and
transported back to the laboratory at the
AMCD Base Station in St. Augustine Beach
where they were centrifuged at 4,375 RPM
for 15 minutes. Samples were then placed
in a labeled and sealed plastic bag, shipped
to the Florida State Department of Health
(FDOH) Virus Laboratory in Tampa, Florida and tested for WN, EEE, and HJ viruses.
Blood samples were sent to the Laboratory
on a Monday and results reported to the
District by the end of the same week. Once
a chicken seroconverted it was removed, destroyed, and replaced with a new bird at its
respective location.
Mosquito Surveillance. Population abundance of adult mosquito vectors were monitored by AMCD using CDC light traps (John
W. Hock Company, Gainesville, FL) baited
with octenol (BioSensory, Inc., Putnam, CT)
at 41 permanent locations in the County.
Traps were placed outdoors at the beginning
of March through the last week of November.
Traps were suspended 1 m above ground surface by a shepherd’s hook and operated for
18-20 h using a 12-v battery. Mosquito collections were transported from the field to the
AMCD facility and identified to species using the taxonomic keys of Darsie and Ward
(2005). Monthly mean rainfall and tempera-
15
ture were obtained for the time period 20082014 from NOAA weather station records
(http://www.weathersource.com).
Human Surveillance. Human blood samples obtained by local physicians, and other
health related professionals, were sent to
the FDOH Virus Laboratory in Tampa and
tested for WNV, EEE, and Saint Louis encephalitis virus. Positive results were shared
by St. Johns County Department of Health
with AMCD to determine appropriate local
vector control measures.
Other Animal Surveillance.Dead birds reported by local residents to AMCD were
referred to the St. Johns County Health Department who sent the remains to the FDOH
Tampa Laboratory for detection of WNV.
Local veterinary reports and clinical samples
from horses and dogs with suspected WNV
or EEE infections were tested by the Florida
Department of Agricultural and Consumer
Services (FDACS) Arbovirus Laboratory in
Kissimmee, Florida. This agency shared test
results with St. Johns County FDOH and
AMCD.
Statistical analysis. Simple logistic regressions were used to determine if significant
correlations (a=0.05) existed between sentinel chicken seroconversion events, mean
monthly temperature, and total monthly
rainfall using the statistical software program R (ver. 3.13) (R Core Team 2012).
III. RESULTS AND DISCUSSION
A total of 389,899 mosquitoes were collected from CDC light traps from 2008 to
2014 with an average of 5,026 mosquitoes
collected per site annually. Forty mosquito
species were collected during the study period, including Culiseta melanura Coquillett,
Culex nigripalpus Theobald, and Cx. quinquefasciatus Say, the known vectors of EEEV, HJ
virus, SLE, or WNV.
The only human case of arbovirus infection in St. Johns County from 2008-2014 was
a WNV asymptomatic blood donor reported
by FDOH in September 2014. During 20082014 there were 184 positive WNV, 113 positive EEE, and 34 positive HJ sentinel chickens reported within the County (Fig. 1). No
16
Figure. 1. Annual totals of positive sentinel chicken arbovirus seroconversions for West Nile, eastern equine
encephalitits, and Highlands J virus (2008-2014), St. Johns County, Florida.
arboviral positive dead birds were reported
during this time period. There were 5 confirmed EEE horse cases from 2008 to 2014.
West Nile virus seropositive sentinel
chickens were reported from May through
December with the majority of cases occurring in September (Fig. 2). Eastern equine
encephalitis seropositive sentinel chickens
were reported from March through November with most seroconversions occurring
Figure. 2. Monthly totals of positive sentinel chicken arbovirus seroconversions for West Nile and eastern equine
encephalitits virus (2008-2014), St. Johns County, Florida.
Weaver et al.: Arbovirus Surveillance Report in St. Johns County, Florida, 2008-2014
in June (Fig. 2). Highlands Jay infection of
sentinel chicken flocks was recorded from
April through September. Most seroconversions occurred during July. Eastern equine
encephalitis horse cases occurred in March
through July.
The average rainfall from 2008 to 2014
was 116.9 cm. In 2009, the average rainfall
was 155.1 cm, which was the most rain received in a year during this 7 year period. Total monthly rainfall did have a significantly
positive relationship with EEE serocon rates
in sentinel chicken flocks (β = 0.09, SE =
0.04, p = 0.028, α = 0.05). It appeared that
rainfall may have had a slight negative correlation with WNV (β = -0.07, SE = 0.04, p =
0.06, α = 0.05) and HJ (β = -0.01, SE = 0.06, p
= 0.82, α = 0.05) seroconversion in sentinel
flocks but these relationships were not statistically significant.
The average temperature from 2008 to
2014 was 21.0°C. The highest monthly average temperature was recorded at 21.7°C in
2012. For the 2008-2014 time period, the
incidence of WNV in sentinel chickens was
negatively correlated with average monthly
temperature (β = -0.13, SE = 0.02, p = 1.78e-8,
α = 0.05) while EEE seroconversions were
positively correlated with temperature (β =
0.11, SE = 0.02, p = 4.71e-6, α = 0.05). There
was no significant relationship between HJ
and temperature (β = 0.07, SE = 0.04, p =
0.05, α = 0.05).
Arbovirus transmission is influenced by a
complex system of environmental, host, and
vector interactions (Tabachnick, 2010). Despite the complex nature of this system, the
combination of surveillance methods used
by AMCD provides the basic framework necessary to protect the health and well being
of citizens and tourists alike. However, we
suggest that future agency surveillance and
17
geographical sampling resolution can be
strengthened through the adoption of novel
mosquito arbovirus surveillance technologies incorporating geographic information
systems (GIS) and molecular diagnostics.
IV. ACKNOWLEDGEMENTS
We thank all employees of AMCD and
former employees, Whitney Qualls and Ali
Fulcher, who partially coordinated and participated in this program. We are thankful
for the support from the property owners
who allowed us to use their property to place
the sentinel chickens.
VI. REFERENCES CITED
Connelly, C.R., J.F. Day, and W.J. Tabachnick. 2012.
West Nile Virus. Institute of Food and Agricultural
Sciences (IFAS), University of Florida, Gainesville,
FL. Pub. #ENY-642.
Darsie, R.F. and R.A. Ward. 2005. Identification and Geographical Distribution of the Mosquitoes of North America,
North of Mexico. University of Florida Press, Gainesville, FL.
Hayes, E.B., J.J. Sejvar, S.R. Zak, R.S. Lanciotti, A.V.
Bode, and G.L. Campbell. 2005. Virology, pathology, and clinical manifestations of West Nile virus
disease. Emerg. Infect. Dis. 11:1174-1179.
O’Bryan, P.D. and H.J. Jefferson. 1991. The year of the
chicken: the good and bad of a sentinel chicken
flock during the 1990 Florida SLEV epidemic. J.
Florida. Mosq. Control Assoc. 62:59-63.
R Core Team. 2012. R: A Language and Environment for
Statistical Computing. Version 3.13. R Foundation
for Statistical Computing, Vienna, Austria. ISBN
3-900051-07-0.
SJC TDC. (St. Johns County Tourist Development
Council) 2015. Tourism in St. Johns County. St.
Johns County, Florida, Government. http://www.
co.st-johns.fl.us/TDC [Accessed 2 June 2015].
Tabachnick, W.J. 2010. Challenges in predicting climate and environmental effects on vector-borne
disease episystems in a changing world. J. Exp. Biol.
213:946-954.
Xue, R.D. and W.A. Qualls. 2008. Arbovirus surveillance
in St. Johns County, Florida. Tech. Bull. Florida
Mosq. Control Assoc. 8:27-30.
SPATIAL ANALYSIS OF ARBOVIRUS TRANSMISSION
IN ST. JOHNS COUNTY, FLORIDA
MOHAMED F. SALLAM1,2,3, CATHERINE LIPPI1, AND RUI-DE XUE1
1
2
Department of Entomology and Nematology, University of Florida
Gainesville, FL 32611
Department of Entomology, College of Science
Ain Shams University
Cairo, Egypt
3
ABSTRACT. The arbovirus surveillance program conducted by Anastasia Mosquito Control District is an early warning monitoring
system designed to identify the factors affecting the transmission risk of vector borne diseases. The sporadic frequency and intensity
of eastern equine encephalitis virus (EEEV)
and West Nile virus (WNV) transmission in
St. Johns County prompted the use of spatial
pattern analysis to understand the geographic distribution of both diseases in the County
using Geographic Information System (GIS)
technology. The significance of GIS application in surveillance programs stem from its
potential to determine the spatial distribution patterns and directional trends of diseases and to produce transmission probability
risk maps. Data from 2008-2014 on frequency
and intensity of EEEV, WNV sentinel chicken
seroconversions and subsequent mosquito
vector abundance of Culiseta melanura, Culex
nigripalpus, and Cx. quinquefasciatus were used
in analyses. Geographically, mosquito vector
density and frequency/intensity of disease
transmission was randomly distributed. This
was also confirmed by logistical analysis that
indicated a significantly reduced dependency
of EEEV and WNV chicken serconversion records with vector density.
I. INTRODUCTION
In the past decade, the adoption of Geographic Information System (GIS) technologies by public health agencies has promoted
an increased awareness of the spatial and
temporal distributions of disease. A powerful tool in the management of community
level diseases, geospatial methods have been
successfully used to model disease transmission dynamics and produce spatial predictive
maps of disease distribution (Dambach et al.
2012). Geographic Information Systems have
also proven useful for studying the spatial and
temporal distribution patterns of mosquitoborne diseases and their vectors (Kulkarni et
al. 2010, Abdel-Dayem et al. 2012, Sallam et
al. 2013). The interactions between environmental variables, hosts, and disease vectors
are often complex, so the ability to accurately model these parameters at the local level
would provide valuable information for public health management decisions, policy formation, surveillance, and disease mitigation.
The Anastasia Mosquito Control District
(AMCD) currently provides all mosquito
surveillance and control in St. Johns County,
Florida, with the goals of mitigating disease
burden and minimizing transmission risk. St.
Johns County lies in the northeastern part of
Florida, USA with a total area of 1,588 km2
(609 sq. miles). The County is positioned
between the St. Johns River to the west and
the Atlantic Ocean to the east (Figure 1). As
a result, there is great diversity in saltwater,
Key Words. eastern equine encephalitis virus, West Nile virus, Culiseta melanura, Culex
nigripalpus, Culex quinquefasciatus, Geographic Information System
18
Sallam et al.: Spatial Analysis of Arbovirus Transmission in St. Johns County, Florida
19
Figure 1. Culex quinquefasciatus collection sites within 5 km buffer zones around sentinel chicken traps.
freshwater, and vegetative habitat features
throughout the County, providing developmental habitats for about 43 mosquito species
(Naranjo et al. 2014). Eleven of these species
are known vectors of several pathogens, notably arthropod-borne viruses (Lord and Day
2001, Sardelis et al. 2001, Blackmore et al.
2003, Cupp et al. 2003). Public health con-
20
cern has continued to surface over mosquitoborne diseases since the first domestic case of
West Nile virus (WNV) was recorded in New
York in 1999 (Hayes et al. 2005). West Nile
virus was first reported in St. Johns County in
2001 (Connelly et al. 2012).
Arbovirus surveillance in Florida serves
as an early warning system for potential
outbreaks of mosquito-borne viral diseases.
Established in 1978, the Arbovirus Surveillance Network is the primary system for
monitoring mosquito-borne diseases in
the state of Florida (O’Bryan and Jefferson
1991). This network is largely reliant on
the use of sentinel chicken flocks to monitor endemic virus transmission. In St. Johns
County, sentinel chicken arboviral surveillance is a core function of AMCD. Sentinel
chicken seroconversion records and mosquito surveillance data are collected by the
District that, in turn, aids public health and
mosquito control officials in assessing the
incidence and transmission risk of endemic
virus transmission.
Data driven conclusions drawn from
surveillance data provide the necessary justification for the application of larvicides
and adulticides. Prior to our study, there
has been no attempt to quantify the spatial
distribution of WNV and EEEV in St. Johns
County. In this report, we present a spatial
analysis of the AMCD’s mosquito arbovirus
surveillance program for EEEV, WNV, and
abundance of vectors in St. Johns County
for the years 2008–2014. The delineation of
geographic distribution patterns and spatial direction trends of both diseases and
their vectors may help to gain insight into
their spatial characteristics. Indeed, mosquito abundance has been addressed in
previous studies by other authors and have
been identified as a major predictor in the
likelihood of epizootics (Scott et al. 1983,
Anderson et al. 1999, Lord and Day 2001).
Also, we plan on comparing EEEV and
WNV seropositive sentinel chicken records
with that of vector population density in order to test the hypothesis that intensity of
EEEV and WNV transmission are correlated
with the density of the mosquito vectors in
St. Johns County.
Sentinel Chicken Surveillance. Sixty, 21week old chickens were distributed over 12
locations in the County at the beginning of
April through December every year during
2008-2014 to monitor arbovirus activity. Approximately 2.0 ml of blood was taken from
each chicken’s wing vein once a week. Blood
samples were kept in labeled vacutainers
(Fisher Scientific) and transported back to
the laboratory at the AMCD base station in
St. Augustine Beach where they were centrifuged at 4,375 RPM for 15 minutes. Samples
were then placed in a labeled and sealed
plastic bag, shipped to the Florida State Department of Health (FDOH) Virus Laboratory in Tampa, Florida where blood samples
were tested for WNV, and EEEV. Samples
were sent to the Lab on Mondays and results
reported to AMCD by the end of the same
week. Once a chicken seroconverted it was
removed, destroyed, and replaced with a
new bird at its respective location.
Mosquito Surveillance. Population abundance of adult host seeking mosquito vectors were monitored by AMCD using CDC
light traps (John W. Hock Company, Gainesville, FL) baited with dry ice at 68 permanent
locations in the County. Traps were placed
outdoors at the beginning of March through
the last week of November during 2008-2014.
Traps were suspended 1 m above ground surface by a shepherd’s hook and operated for
18-20 h using a 12-v battery. Mosquito collections were transported from the field to
the AMCD facility for further identification
to species level using the taxonomic keys of
(Darsie and Ward 2005).
Abundance data of WNV Culex quinquefasciatus Say and Cx. nigripalpus Theobald,
and EEEV vectors, Culiseta melanura (Coquillett), were extracted from AMCD field
records within a 5 km buffer zone around
each sentinel chicken trap using the geoprocessing package in ArcGIS (ver. 10.0). Buffer
zones were used as an indication of probable mosquito transmission within their flight
range around sentinel chicken traps (DeMeillon 1934, Nayar and Sauerman Jr 1973)
(Figures 1-2). In addition, the study area was
21
Figure 2. Culiseta melanura collection sites within 5 km buffer zones around sentinel chicken traps.
categorized into AMCD operational route
zones (Figure 3) previously extracted from
St. Johns County GIS division. This was
done in order to provide data driven guidance risk maps for the operational surveil-
lance and control activities conducted by
AMCD field personnel. In addition, these
operational route zones were coupled with
the risk maps produced in order to accurately identify the focal risk areas.
22
Figure 3. Map of St. Johns County showing operational route zones and open water bodies.
Spatio-Statistical Analysis. Parameters of
EEEV and WNV seroconversion data (frequency and intensity), and mosquito vector
density were analyzed using spatial statistics
tools in ArcGIS (ver. 10.0). The standard de-
viational ellipse (SDE) analyses in this package were used to summarize the spatial patterns of the above parameter values around
their mean. The SDE creates a polygon to
summarize the spatial directional distribu-
tion which may show dispersion, central
tendency, and directional trends. Therefore,
SDE was used to map the directional distribution trend of mosquito vectors and spread
of EEEV and WNV transmission.
Spatial autocorrelation analysis, using
Global Moran’s I Index, was used to identify
the distribution pattern of mosquito vectors,
EEEV, and WNV sentinel chicken seroconversion. The spatial autocorrelation pattern
may be random, clustered, or dispersed (Lee
and Wong 2001). The distribution pattern is
predicted by comparing values of neighboring sampling sites for: 1.) mosquito vector
density and 2.) frequency and intensity of
EEEV and WNV seroconversion records. A
strong positive spatial autocorrelation (clustered) is indicated when the neighboring
sampling points have similar values. Whereas, if the neighboring sampling points have
dissimilar values (dispersed), then it indicates strong negative spatial autocorrelation.
The value of Moran’s I Index is between -1
and 1. The ‘Z’ score value is calculated to test
whether the observed clustering or dispersing is significant. When the Z score indicates
statistical significance, a positive Moran’s I
index indicates tendency toward clustering
while a negative value indicates tendency toward dispersion. When the Z score value is
not significantly different from 0, there is no
spatial autocorrelation and the pattern does
not appear to be significantly different from
a random distribution. The null hypothesis
(Ho) for the current analysis was that there
is no spatial clustering of EEEV, WNV, with
abundance of the mosquito vectors in St.
Johns County.
Logistic regression R statistical software
(ver. 3.13) was used to assess the relationship
between frequency of sentinel chicken seroconversion records and the abundance of
their corresponding mosquito vectors (i.e.,
Cs. melanura, Cx. nigripalpus, and Cx. quinquefasciatus) sampled within a 5 km buffer zone
around the chicken flocks.
From 2008 through 2014 there were 113
and 184 sentinel chicken seroconversions re-
23
ported from the County for EEEV and WNV
antibodies, respectively. A total of 389,899
mosquitoes were collected from CDC light
traps with an annual average of 5,026 per
site. Forty mosquito species were collected
during the study period including EEEV and
WNV bridge vectors and other non-anopheline mosquitoes that represented 68.43%
of the total mosquitoes collected. Culiseta
melanura, Cx. nigripalpus, and Cx. quinquefasciatus represented 1.01%, 5.91%, and 2.72%,
respectively of the total non-anopheline
mosquitoes collected during the study.
The SDE analysis indicated that density
of Cs. melanura, Cx. nigripalpus, and Cx. quinquefasciatus and intensity records of EEEV,
WNV were randomly distributed and showed
a directional trend about the mean; this is
verified by the ellipsoid polygons in Figures
4-6. SDE analysis revealed that a random
distribution pattern highlighted the habitat
suitability for these mosquito vectors within
the ellipsoid polygons. These results were
further confirmed by Moran’s I index values
where these same parameters were found
to be insignificant (P>0.05) confirming the
findings generating by the SDE polygons.
Because we sampled adult host seeking mosquitoes, these individuals were most likely
newly emerged from their developmental
habitats within the risk polygons. Accordingly, this reflects the probable abundance
of larval habitats for these vectors within the
polygons. Similarly, extracted data on mosquito vectors around sentinel chicken traps
reflected the flight range of these individuals and the availability of their host(s) as a
source for blood meals. Although adult host
seeking mosquito vectors were collected outside the risk polygons (which may have been
infective with either EEEV or WNV) larval
habitats within the ellipsoid polygons are
worthy of surveillance and should be evaluated for control compared with other areas.
The random distribution of seropositive
records of EEEV and WNV indicated virus
circulation inside the mosquito vectors rather
than virus amplification inside bird host(s).
Although the spatial analysis potentially predicted the directional distribution pattern of
infective mosquito vectors, and their flight
24
39.4%
42%
61%
Figure 4. Statistical distribution pattern of WNV and Culex quinquefasciatus in St. Johns County showing standard
deviational ellipsoids.
range, during host seeking, the number of
parameters used in this analysis prevented us
from highlighting virus amplification inside
their reservoir host(s). This limitation is at-
tributed to lack of consistent distribution data
on reservoir host(s), which represents the
source of infection for uninfected mosquito
vectors within their flight range.
25
42%
51%
39.4%
Figure 5. Statistical distribution pattern of WNV and Culex nigripalpus in St. Johns County showing standard
The GIS spatial analysis predicted high risk
areas for mosquito bites for Cs. melanura, Cx. nigripalpus, and Cx. quinquefasciatus that covered
806.67 km2 (50.80%), 825.12 km2 (51.96%),
and 961.16 km2(60.53%) of the total land
area of the County, respectively. These areas
were found to represent AMCD operational
route zones N01-N03, C04-C08, S01-S03 for
the three mosquito vectors, respectively. Furthermore, the amount of land under EEEV
26
51%
45%
20.29%
Figure 6. Statistical distribution pattern of EEEV and Culiseta melanura in St. Johns County showing standard
and WNV transmission risk from probable
infective mosquito vectors represented 45%
and 42%, respectively, of the total area of St.
Johns County. These risk areas were found to
represent operational route zones N02, N03,
C04-C08, S01 for EEEV and N02, N03, C04C09, S01, S02 for WNV. Overlapping between
polygons produced for high transmission risk
areas shown in Figs. 4-6 show the likelihood
of probable association between pathogen,
vector, and reservoir host. In this context,
the overlapping areas under risk of probable
infective mosquito bites should be targeted
for EEEV and WNV surveillance and control
activities rather than the entire county.
The predicted distribution developed by
SDE polygons demonstrated the relative expanded distribution pattern of Cx. quinquefasciatus compared with the other two vectors. This reflected either a wide flight range
of this mosquito or the availability of suitable
larval habitats and/or availability of hosts as
a blood meal source. However, the parameters used in our analysis prevented us from
addressing actual flight range away from
their larval habitats. Further investigations
are needed to include more parameters on
larval developmental sites in proximity to
wild reservoir host populations.
In previous studies, the density of mosquito vectors played a major role in predicting the transmission of arboviral diseases
27
(Scott et al. 1983, Anderson et al. 1999,
Lord and Day 2001). We found that eastern equine encephalitis virus seroconversion rates were significantly correlated with
the density of Cs. melanura (β = 0.001, SE =
0.000, p = 1.10e-7, α = 0.05) (Figure 7). Also,
the WNV serocons were positively correlated
with the abundance of Cx. quinquefasciatus (β
= -0.002, SE = 0.000, p = 5.26e-10, α = 0.05)
and Cx. nigripalpus (β = 0.003, SE = 0.000, p
= 1.10e-14, α = 0.05) (Figure 8). Surprisingly,
even though correlations were significant
(i.e. P<0.05) between seroconversion rates
and vector abundance, in our study, the dependency of seroconversion on mosquito
vector density was considerably reduced because both values of (β) and (p) were close
to zero which indicated that those relationships were random. These results highlight
the important contribution that other culicine bridge vector(s) may have in the disease
transmission cycle. Moreover, our findings
Figure 7. Annual abundance of eastern equine encephalitis virus seropositive sentinel chicken records, Culiseta
melanura density during 2008-2014.
28
Figure 8. Annual abundance of West Nile virus seropositive sentinel chicken records, Culex. quinquefasciatus, and
Cx. nigripalpus density during 2008-2014.
were confirmed by the evident slight shifting
in SDE polygons between mosquito vectors
and their diseases (Figures 4-6). Previous
studies confirmed that Cs. melanura, Cx. nigripalpus, and Cx. quinquefasciatus are competent vectors in transmitting EEEV and WNV
(Anderson et al. 1999, Blackmore et al. 2003,
Cupp et al. 2003) in comparison with other
mosquito vectors. However, the contribution
of other culicine bridge vectors such as Cx.
restuans Theobald, Cx. erraticus (Dyar and
Knab), Aedes vexans (Meigen), and Uranotaenia lowii Theobald, for EEEV transmission
(Chamberlain et al. 1954, Cupp et al. 2003,
Cohen et al. 2009, Estep et al. 2013) and Cx.
salinarius Coquillett for WNV (Sardelis et al.
2001, Blackmore et al. 2003) should be considered. The contribution of other bridging
vectors exacerbates the complexity of virus
circulation in nature. The variation of feeding preference by bridge vectors may sustain
virus circulation in other reservoir host(s)
such as amphibians and possibly reptiles
(Cupp et al. 2003, Cupp et al. 2004).
In summary, AMCD arbovirus surveillance showed that the overall distribution
of EEEV, WNV and their mosquito vectors
were confined to certain areas of St. Johns
County. This may be attributed to possible
ecological niches for mosquito vectors, seasonal abundance of animal reservoirs such
as birds, amphibians, reptiles, and possible
bridge vector(s). Additional information is
needed from these parameters in order to
shed light on virus amplification and characterize various habitats suitable for mosquito vectors and reservoir host(s). Although
some disease surveillance methods provide
more information than others, combining
traditional operational surveillance programs with GIS technologies and modeling
packages will enhance the ability of public
health agencies to maximize the benefits of
practical application of disease surveillance
data. Developing real time data driven risk
maps will help in the decision making process by prioritizing control activities, at the
District level, while increasing control effectiveness, plus reducing labor and associated
cost especially during periods of arboviral
outbreaks.
We would like to express our deep gratitude to Mr. Richard Weaver for his consistent help in supporting us with the necessarily data for the current work. Also, we extend
our thanks to all employees in AMCD and
former employees especially Dr. Whitney
Qualls and Ali Fulcher. Moreover, authors
extend their gratitude to the two anonymous
reviewers for their valuable comments that
potentially helped in improving the readability and the significance of the current
investigation.
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RELATIONSHIP BETWEEN CITIZEN KNOWLEDGE,
VEGETATION COVERAGE, AND FREQUENCY of
REQUESTS FOR MOSQUITO CONTROL SERVICE IN ST.
JOHNS COUNTY, FLORIDA
CLAUDIA A. DAVIDSON1, JODI M. SCOTT2, TANJIM H. HOSSAIN3,
JOHN C. BEIER1, 3, AND RUI-DE XUE2
1
Department of Public Health Sciences, Miller School of Medicine
University of Miami, Miami, FL 33136
500 Old Beach road, St. Augustine, FL 32080
2
Leonard and Jayne Abess Center for Ecosystem Science and Policy
University of Miami, Coral Gables, FL 33146
3
ABSTRACT. Customer service requests play
an important role in mosquito control programs by providing mosquito surveillance
information and by allowing the opportunity for inspectors to educate the public on
mosquitoes and mosquito borne diseases. In
this study, we assessed differences in knowledge, attitude, mosquito control practices
(KAP), and vegetation characteristics between frequent service requesters and nonfrequent service requesters by conducting
paper, vegetation, and entomological surveys. There were no significant differences
in customer knowledge about mosquitoes
between the two groups. However, there was
a general lack of knowledge about mosquito
resting and sugar feeding behavior in >90%
of participants. Perceptions of mosquito infestation and vegetative coverage were determined to be driving factors for frequent
service requests. Frequent service requesters
reported being bothered or bitten by mosquitoes a few days a week or daily more often than non-frequent; demonstrating that
perceptions of the severity of mosquito infestation are associated with service request
volume. Vegetation coverage on residential
properties of frequent service requesters was
13% higher than non-frequent service requesters and identified vegetation coverage
as another potential factor driving service request volume. We conclude that future edu-
cation programs should include information
on the associations of plants with mosquito
behavior, such as resting and sugar feeding,
to better inform the public and aid in increasing their knowledge base as they relate
to homeowner control practices.
Key Words. Service request; Vector Control
Management System; KAP survey
I. INTRODUCTION
Although the majority of mosquito-related issues in the United States arise from them
being a nuisance, there remains an imminent
threat of the emergence and re-emergence of
mosquito-borne diseases. In 2002, 4,156 cases
of the West Nile Virus were reported in the
United States; in 2005, 25 cases of dengue fever were reported in Texas; and between 2009
and 2010, 90 cases of dengue fever cases in
Key West, Florida were reported (O’Leary et
al. 2004; CDC 2007, CDC 2013). By the end
of 2014, there have been 11 locally acquired
chikungunya cases in Florida, and though it
rarely results in death, the disease can be debilitating to those infected (CDC 2014).
Mosquito control programs, like Anastasia Mosquito Control District (AMCD) of
St. Johns County, Florida, use surveillance
methods to monitor nuisance and vector
30
Davidson et al.: Relationship Between Citizen Knowledge, Vegetation Coverage, and Frequency
capable mosquitoes to reduce and prevent
the spread of mosquito-borne diseases (Day
and Shaman 2011). Despite the efforts of
these programs, an increase in the incidence
of mosquito-borne diseases is still of major
public health concern. It is imperative that
these programs continue to conduct applied
research, as well as continued surveillance
to produce new or improved approaches for
mosquito control.
The District uses the Vector Control
Management System (VCMS) to manage
service requests made by residents of St.
Johns County. Service requests entered
into VCMS play a major role in supporting
surveillance efforts (AMCD 2013, Weaver
et al. 2013). This method of surveillance
has proven to be successful as there is a
correlation between the volume of service
requests received and the density of mosquitoes (Morris and Clanton 1981, 1992).
The software allows for service requests to
be linked with mosquito abundance, along
with information of their location in the
county. The VCMS system relies on people
to make service requests; however, some
residents make more frequent requests
than others. Moreover, there seems to be
a disconnect in the knowledge, attitude,
and control practices between residents
that call in service requests frequently and
residents who do not (AMCD staff pers.
comm.).
In addition, over the last few years,
AMCD staff has observed that residents who
frequently put in service requests seem to
have more vegetation on their property than
residents without service requests (AMCD
staff pers. comm.). The density of vegetation
on residential properties is important because mosquitoes rely on sugar from plants
for energy (Hocking 1953, Nayar and Van
Handel 1971, Van Handel 1965) and vegetation for resting sites (Allan et al 2009).
Energy gained from plants is used to support their host seeking behavior in flight
and fecundity, increasing the probability of
disease transmission (Muller et al. 2011, Yuval 1992). Therefore, the method in which
a residential property owner manages his/
her agricultural landscape is correlated with
31
mosquito population abundance (Day and
Shaman 2011).
Understanding residents’ knowledge and
attitude, toward mosquito behavior and ecology on residential properties is essential for
developing and/or improving mosquito control approaches. Therefore, the purpose of
this study is to: 1) determine if there are differences in knowledge, attitude, and control
practice of residents in regards to mosquitoes
between residents; 2) determine if service requests are related to vegetation on properties;
3) identify major vegetation density on residential properties of service requesters; and
4) determine the major species of mosquitoes
and abundance in residential properties. This
study will provide feedback to AMCD and
lessons learned can be used as a tool by the
District to continue to protect public health
by revising educational materials, and serving
as a catalyst for discussion on future studies
involving vegetation, mosquito behavior, and
control.
Study site selection and sampling. Data collection was conducted in St. Johns County,
Florida between June 3 and July 31, 2013. St.
Johns County, with a population of approximately 202,000, covers 609 square miles
of northeastern Florida (US Census 2010,
Weaver et al. 2013). The VCMS is an integrated database and mobile field data collection system used by AMCD for mosquito
control (Weaver et al. 2013). The software
assists with management of the District’s
mosquito borne disease and surveillance
data; integrating mosquito management
activities (service requests, inspection, and
spraying); and monitoring employee activities. Using the VCMS program, four years’
worth of service requests was assessed to categorize and identify residential properties.
Service requests were grouped as 1, 2, 3, 4,
and >4 requests by household within each
year. The data was filtered by deleting multiple repeat entries that were not different
requests, this allowed for a more accurate
description of the frequency of calls over the
past four years.
32
Residential properties that had frequent
service requests were selectively sampled
from the VCMS database. Requests had to
meet three criteria: 1) be a residential property in which an occupant has made >3 requests in 2012; 2) multiple requests had to
have the same address but different dates of
entry; and 3) properties could not be farm
lands, apartment complexes, or greater than
43,560 sq ft. If occupants were not available
at the pre-selected household, then the next
pre-selected household was visited until consent was granted.
Residential properties that were not frequent service requesters or non-service requesters, were selected at random as control
sites. Control sites also had to meet three criteria: 1) residential properties had to be within
the same neighborhood as a study site, either
on the same street or the next street over; 2)
they were not identified as having >3 service
requests a year between 2009-2012; and 3)
properties could not be farm lands, apartment complexes, or a larger than≥43,560 sq
ft. If occupants of a household were not available or refused to participate, neighboring
houses were visited until consent was granted.
Study sites identified with these criteria and sampling techniques resulted in 24
residential homes within 4 zip codes: 32080,
32082, 32084, and 32086. Of these study
sites, six houses were randomly selected
from each zip code, three of which were
identified as frequent service requests from
the VCMS database and three randomly selected control sites. All houses were visited
during the evening hours (4pm-6pm) for 1.5
weeks: July 8-16, 2013.
Paper-based KAP survey. A member of the
research team administered a knowledge,
attitude, and practice (KAP) questionnaire
to one consenting adult (≥18 years old)
from each household. The questionnaire
was designed to elicit resident’s knowledge
on mosquito behavior, mosquito-borne disease, attitudes towards and practices used in
mosquito control. Their responses were assumed representative of the household. All
questionnaires were asked face-to-face and
most questions were asked as open-ended
questions. No demographic information
was collected, as it was not necessary for the
completion of this study.
Vegetation survey. After each questionnaire was administered, consent was requested to survey their property and measure
vegetation growth. This measurement was
taken with the use of a 1,000 ft tape-measuring wheel (#PSMW48, Lufkin, Sparks, MD).
All vegetation above ground level was measured by taking the circumference or perimeter and calculating its area in square feet.
Tree canopies and grass coverage were not
measured as investigators were only interested in non-grass vegetation that covered
the soil surface. Circumference of trees was
measured at their base and included in the
survey. To calculate vegetation coverage of
the property, measurements were summed
and divided by the total square footage of
the property. Square footage of each property was obtained from the St. Johns County
Property Appraiser’s Office website. Photos
of vegetation were taken on each property.
Frequently occurring plants on each property were identified by Ali Fulcher, at AMCD,
and Keith Fuller, at the St. Johns County Agriculture Center.
Entomological survey: In addition to
the vegetation survey, consent to check participants’ yards for adult mosquitoes and
mosquito larval habitats were also obtained
upon completion of the questionnaire. To
determine the presence of adult mosquitoes,
landing rate counts (LRC) were conducted
by volunteers. Landing rate counts counts
consisted of volunteers exposing the surface
from their elbow to their hand, and counting how many mosquitoes landed, within
three minutes at each property. Mosquitoes
were allowed to land, but were not allowed
to take a blood meal. Landing rate counts
were conducted in areas identified as possible resting areas (shaded vegetated areas).
Each property was also checked for containers or plants with standing water for the
presence or absence of larvae or pupae. Larvae and pupae were not collected. Containers
found with standing water were emptied by
surveyors. Adult mosquitoes were collected
with the use of a BioGents sentinel (BGS)
trap with BGS lure and were set up for 24 hrs
on each property. The BGS trap was chosen
due to its ability to attract various container
ovipositing mosquitoes such as Aedes spp.
and Culex spp. Mosquito collections were
taken within a day or two of administering
the questionnaire. Collected mosquitoes
were brought back to the lab and kept frozen in a standard freezer until they could be
sexed, identified to species, and counted.
Data analysis. SPSS (ver. 21) was used to
assess differences in vegetation coverage,
knowledge, attitude, and practice between
frequent and non-frequent service requesters. Chi-square tests were used to test for
statistical significance. Mean differences of
vegetation coverage, mosquito collection,
knowledge, and practice scores were calculated using an independent t-test.
III. RESULTS
Paper-based survey. Twenty-four participants completed the survey, 12 of which
were from households that had requested
services frequently and 12 from households
that had not requested services frequently.
Each participant answered every question
on the survey.
Knowledge. An equal number of frequent
service requesters and non-frequent service
requesters (4 each) scored as being very
knowledgeable of mosquitoes. The remaining frequent and non-frequents service requesters (8 each) scored as not being very
knowledgeable about mosquitoes (Table 1).
Participants who rated their propensity of
knowledge as 4-5, on average, had a higher
33
knowledge score than those who self-rated as
being not very knowledgeable.
Overall, 92% of the participants were
able to identify at least one source of mosquito larval habitat such as standing water,
marshes, swamps, or damp/moist areas.
More than 75% of the participants were able
to list at least one mosquito-borne disease,
such as malaria, dengue, West Nile virus,
eastern equine encephalitis.. When asked
where mosquitoes generally rested as adults,
only 25% of participants were able to identify
vegetation as a resting area. Specific knowledge of non-blood meal sources for mosquitoes was low, with only one responder reporting knowledge that plants are a meal source
for mosquitoes. There were no significant
differences between non-frequent service requesters and frequent service requesters in
overall knowledge scores or specific knowledge about mosquito behavior and disease
(Table 1).
Attitude. A majority of the participants
reported being bothered or bitten by mosquitoes daily or a few days a week with 71%
reporting being bothered or bitten daily during the summer months (Table 2). An additional 8% reported being bothered or bitten
a few days a week. There was a significant association (χ2, p = 0.019) between service request status and reporting how often they are
bothered or bitten by mosquitoes. On average, frequent service requesters reported being bothered or bitten by mosquitoes more
often than non-frequent service requesters.
When asked where they were more likely to
be bothered by mosquitoes, over 80% of the
Table 1. Responses to the open-ended questions on mosquito knowledge.
Non-frequent service
requesters
Frequent service
requesters
Overall
Question
n
%
n
%
n
%
Mosquito breeding sources
Mosquito resting area
Mosquito meal sources
Mosquito sugar feed on plants
Mosquito borne diseases
10
2
1
1
10
83.3
16.7
8.3
8.3
83.3
12
4
0
0
9
100.0
33.3
0.0
0.0
75.0
22
6
1
1
19
91.7
25.0
4.2
4.2
79.2
Average Knowledge Score
Mean (SD)
Non-frequent service
requesters
Frequent service
requesters
Overall
2.00 (1.04)
2.08 (0.67)
2.04 (0.85)
34
participants reported being bothered outdoors on their property. When asked what
time of the day they were outdoors on their
property, most participants reported being
outdoors mostly in the evening hours and
multiple times to most of the day (80%).
More than 80% of participants reported
great concern towards reducing mosquito
larval habitats, scoring 4 or 5 on a five point
scale (Table 2). On average, frequent service
requesters reported spending more time
outdoors (22.75 hours) than non-frequent
service requesters (18.67 hours).
Practice. Most participants reported that
they did something to reduce larval habitats,
including all of the frequent service requesters (Table 3). Participants varied in their responses on the measures they take to reduce
larval habitats, with emptying standing water
as one of the most frequent methods used.
Participants also varied in their responses on
the personal protection measures used to
protect themselves and their family, with the
use of mosquito repellent as one of the most
frequently used methods. On average, frequent service requesters had a significantly
higher practice score (p = 0.023) than nonfrequent service requesters (Table 3).
Vegetation survey. Vegetation type and coverage varied between residential properties
(Table 4). While not significant, frequent
service requester properties had 13% greater vegetation coverage than non-frequent
services requesters (Figure 1). When comparing mean differences between these two
groups within each zip code, no significant
differences were found in 3 out of the 4
zip codes with. In zip code 32080 there was
a significant (p < 0.05) difference in mean
vegetation coverage between the two groups
(Figure 2). The more vegetation coverage
the more service requests.
Entomological survey. There were no significant differences between frequent and
non-frequent service requesters in the average number of mosquitoes trapped (Figure
4). The majority of the mosquitoes caught
in the BGS traps were container ovipositing
mosquitoes, of which 70% were Aedes albopictus (Skuse). Average landing rate counts,
conducted 3 months post-initial survey, were
less than or equal to one, with the exception
of one home, which had an average count
of 5.7 mosquitoes. Approximately half of the
houses surveyed were positive for containers
with standing water. However, there were no
Table 2. Survey mean responses (SD) to open-ended questions about attitude towards mosquitoes.
Question
How often were you bothered or bitten by mosquitoes?*
Daily or few days a week
Few days a month or fewer
Where on your property you were most likely to be bothered
Outside around house
Inside Home
Everywhere
Not sure
What time of the day outdoors were you bothered?
Morning
Afternoon
Evening
Multiple times/All day
How concerned are you about reducing mosquitoes?
0-3 score, not very concerned
4-5 score, very concerned
Time spent outdoor on property per week
Non-frequent
service
requesters’ n
(%)
Frequent
service
requesters n
(%)
Overall n
(%)
7 (58.3)
5 (41.7)
12 (100.0)
—
19 (79.2)
5 (21.8)
8 (66.7)
1 (8.3)
12 (100.0)
—
20 (83.3)
1 (4.2)
2 (16.7)
1 (8.3)
—
—
2 (8.3)
1 (4.2)
2 (16.7)
1 (8.3)
3 (25.0)
6 (50.0)
2 (16.7)
10 (83.3)
18.67 (13.4)
Percentages are column % out of 12 within each group and out of 24 for overall
*significant difference in response between groups (p < 0.05)
1 (8.3)
1 (8.3)
2 (16.7)
8 (66.7)
1 (8.3)
11 (91.7)
22.75 (14.6)
3 (12.5)
2 (8.3)
5 (21.8)
14 (58.3)
3 (12.5)
21 (87.5)
20.7 (13.9)
35
Table 3. Survey responses to open-ended questions about practices for protection against mosquitoes.
Non-frequent ser- Frequent service
vice requesters
requesters
(n)
(n)
Question
Do you do anything to reduce mosquito breeding sources?
No
Yes
What measures do you take to reduce sources?
Self app. of store bought pesticide
Emptying standing water
Call county services (AMCD)
Other
What personal protection measures do you take to protect you
and your family?
Mosquito repellent
Citronella candles
Wearing long sleeves and long pants
Alter outdoor activities
Other
3
9
0
12
3
21
1
8
3
1
4
11
4
2
5
19
7
3
7
1
0
4
1
6
4
1
4
3
13
5
1
8
4
Non-frequent ser- Frequent service
vice requesters
requesters
Practice score
Mean (SD)
2.92 (1.44)
significant differences in the presence or absence of containers with standing water between the two groups.
IV. DISCUSSION
There were no significant differences in
overall or specific knowledge about mosquitoes between the two groups, where most
participants reported not being very knowledgeable about mosquitoes. Most residents
Overall
(n)
4.25 (1.22)
Overall
3.58 (1.47)
were able to identify mosquito larval habitats and at least one mosquito-borne disease.
However, collectively, there was a lack of
knowledge about mosquito resting and feeding behavior with only six individuals correctly identifying some kind of vegetation
as a mosquito resting area. Only one individual correctly identified plants as a meal
source. The other residents reported that
mosquitoes generally rested on humans and
animals and that they feed on human and/
Table 4. Commonly found plants on St. Johns County residential properties.
Scientific name (family)
Bromelia spp. (Bromeliaceae)
Common name
Bromeliads
Plant description
– Native to tropical America
– Flowering plant
Polystichum munitum (Dryopteridaceae)
Plumbago spp. (Plumbaginales)
Sword fern
Wood fern
Leadwort
Callicarpa spp. (Verbenaceae)
Beauty berry
Tradescantia spp. (Commelinaceae)
Spiderwort
Ruellia spp.
(Acanthaceae)
Petunias
*p < 0.05 the difference between frequent and non-frequent
– Cleft calyx that can collect water and become a
breeding source for mosquitoes
– Found in temperate and tropical regions
– Non-flowering
– Native to South Africa
– Flowering ornamental plant
– Native to east North America and Southeast Asia
– Flowering and fruit producing plant
– Native throughout Gulf Coast region
– Invasive species, used as ornamental plant
– Native from South America
– Flowering plant
36
Figure 1. Average vegetation coverage overall between frequent service requesters and non-frequent
service requesters (±SE).
or animal blood for their main meal source.
The beliefs of the majority of the interviewees, could be due to lack of education and
dissemination about mosquito behavior. By
reviewing educational materials provided
by national and local institutions, it was observed that there was a constant emphasis
on disease information and control/prevention practices for the layman that included
reducing standing water, wearing protective
clothing, and altering daily activities around
dawn and dusk. Although these methods
have been repeatedly proven to be effective
in protecting the public health and the environment (WHO 2014), it is not sufficient in
educating the public on mosquito behavior.
As expected, frequent service requesters
reported being bothered or bitten by mosquitoes more often than non-frequent service requesters. In addition, frequent service
requesters tended to spend more time outdoors and to take a number of measures to
protect themselves and their families from
nuisance and disease carrying mosquitoes;
more so than non-frequent service requesters. This supports their belief that they are
bothered or bitten by mosquitoes more often than the non-frequent requesters are
bitten.
We found that there was a mean difference of 13 % higher vegetation seen in the
frequent service request residence, the lack
of significance was probably due to the extremely small sample size and/or the method used to measure vegetation coverage on
residential properties. Previous work with
vegetation coverage utilized a grid method
in which each grid size is dependent on the
plant shrubs, trees, or flowering. Environ-
Figure 2. Average vegetation coverage by zip codes between frequent service requesters and non-frequent service requesters (±SE).
Figure 3. Average mosquitoes collected over 24-hour
collection period with BioGents sentinel traps baited
with BioGents lure, between frequent service requesters and non-frequent service requesters (±SE).
mental factors such as seasonality of plants
and the density of tree canopies, which may
affect the behaviors of mosquitoes, were
not taken into consideration (Daubenmire
1959).
Although vegetation types varied across
all 24 houses, similarities were seen in the
occurrence of a number of plants that can
support mosquito development, feeding and
resting, such as the Bromelia spp., Polystichum
munitum, Ruellia spp., and Callicarpa spp.
Further studies should investigate mosquito
preferences for types of vegetation, as well
as their uses for them for feeding or resting.
The major species of mosquito caught in
BGS traps was Ae. albopictus. This maybe due
to the anthropogenic changes to landscape
vegetation and artificial uncovered containers, in which this species has been proven to
thrive in. In addition, the time of year that
trapping occurred (i.e. a summer month
with high temperature and precipitation)
provided ideal abiotic factors for development of Ae. albopictus (Paupy et al. 2009).
To our best of knowledge, our study is
unique in its assessment of key factors (e.g.,
KAP and vegetation) driving the volume of
mosquito control service requests. We conclude that knowledge and practice did not
vary greatly by service request volume, but
residents perceptions of how often they are
bothered/bitten was a driving factor for
service request volume. Overall, residents
were proficient in their general knowledge
about mosquito larval habitats and disease;
however, they lacked knowledge about rest-
37
ing and feeding behavior of adult mosquitoes. To be comprehensive, future educational materials for mosquito control and
disease prevention should include this information. Vegetation was not found to be a
driving factor for service request calls. Vegetation cannot be ruled out. Previous studies have shown that mosquitoes do rely on
vegetation for resting sites and plant sugars
for fecundity and flight (Muller et al. 2001,
Yuval 1992). Future research should expand the experimental sample size of survey participants for a more accurate analysis
of the correlation of vegetation and service
requests.
V. ACKNOWLEDGMENTS
Thanks to A. Fulcher, M. Smith, and K.
Fuller for their assistance during this study.
All human subjects were informed before
administering surveys based on the Districtapproved protocol for using human subjects.
Allan, S.A., D.L. Kline, and T. Walker. 2009. Environmental factors affecting the efficacy of bifenthrintreated vegetation for mosquito control. J. Am.
Mosq. Control Assoc. 25: 338-346.
AMCD (Anastasia Mosquito Control District). 2013.
About AMCD [Internet]. http://www.amcdsjc.org/
about-amcd/operations/district.aspx [Accessed August 28, 2013].
CDC (Center for Disease Control and Prevention).
2007. Dengue hemorrhagic fever- U.S.- Mexico border, 2005 [Internet]. Available from the CDC Morbidity and Mortality Weekly Report. 56 (31):785-9
http://www.cdc.gov/mmwr/preview/mmwrhtml/
mm5631a1.htm [Accessed August 29, 2013].
2013. West Nile virus neuroinvasive disease incidence by state [Internet]. Available from CDC
http://www.cdc.gov/westnile/statsMaps/preliminaryMapsData/incidencestatedate.html [Accessed
August 29, 2013].
2014. Chikungunya virus in the United States [Internet].
http://www.cdc.gov/chikungunya/geo/
united-states.html [Accessed September 1, 2014].
Daubenmire, R. 1959. A canopy coverage method of
vegetation analysis. Northwest Sci. 33: 43-64.
Day, J. F. and J. Shaman. 2011. Mosquito-borne arboviral surveillance and the prediction of disease outbreaks. In: Flavivirus Encephalitis. Dr. Daniel Ruzek
(Ed) [Internet]. Available from: In Tech. http://
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Hocking, B. 1953. The intrinsic range and speed of
flight of insects. Trans. R. Ent. Soc. Lond. 104: 223345.
Morris, C. D. and K. B. Clanton. 1981. Significant associations between mosquito control service requests
and mosquito populations. J. Am. Mosq. Control Assoc. 5:36-41.
Morris, C. D. and K. B. Clanton. 1992. Comparison of
people who request mosquito control services and
their non-requesting neighbors. J. Am. Mosq. Control Assoc. 8:65-68.
Müller, G. C., R. D. Xue, and Y. Schlein. 2011. Differential attraction of Aedes albopictus in field to flowers,
fruits and honeydew. Acta Tropica. 118:45-49.
Nayar, J. K., and E. Van Handel. 1971. The fuel for sustained mosquito flight. J. Insect. Physiol. 17:473-481.
O’Leary, D. R., A. A. Marfin, S. P. Montgomery, A. M.
Kipp, J. A. Lehman, B. J. Biggerstaff, V. L. Elko, P.
D. Collins, J. E. Jones, and G. L. Campbell. 2004.
The epidemic of West Nile virus in the United States
2002. Vector-Borne Zoonotic Dis. 4:61-702.
Paupy, C., B. L. Delatte, V. Corbel, and D. Fontenille.
2009. Aedes albopictus, an arbovirus vector: from the
darkness to the light. Microb. Infect. 11:1177-1185.
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J. M. Bequette. 2013. Analysis of Anastasia Mosquito
Control District’s service requests. Wing Beats 24:34-39
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Mosquito species composition and impact
of trapping sites on floodwater mosquitoes,
Aedes Vexans in xinjiang, China
MEI-DE LIU1, YAN-DE DONG1, GUI-LIN ZHANG2, ZHONG ZHENG2,
RUI-DE XUE3, AND TONG-YAN ZHAO1
1
State Key Laboratory of Pathogen and Biosecurity
Institute of Microbiology and Epidemiology, Beijing 100071, P.R. China
2
Center Disease Control and Prevention of Xinjiang Command
Urumuqi, China
3
Abstract. Mosquito species composition
and impact of mosquito trapping sites on a major species of floodwater mosquitoAedes vexans,
was investigated by using Mosquito Magnet
Liberty Plus (MMLP) traps baited with octenol
in Beiwan, Xinjiang. A total of six trapping
sites inside (three) and outside (three) human
occupied areas were selected for the study. Six
species of mosquitoes, Aedes vexans, Anopheles
messeae, Ae. cyprius, Coquillettidia richiardii, Culex
modestus, and Cx. pipiens pipiens were collected
during the study. The results indicated that Ae.
vexans was the major species and represented 97.14% of the total collection. Mosquito
Magnet Liberty Plus traps also significantly
reduced the number of mosquitoes in the residential areas. We found that MMLP traps were
suitable tools for the collection of floodwater
mosquitoes in Xinjiang where extremely high
floodwater mosquito production occurred as
well as provide protection of humans and livestock from host seeking mosquitoes.
nually inundating the residential area and
adjoining semi-desert land. After the flood
water recedes to its normal level, many shallow ponds, lowland puddles, water holes,
and swamps become ideal production sites
for floodwater mosquitoes. Previous studies,
have shown that extremely high floodwater
mosquito production occurs creating a serious health threat for humans and livestock
(Lu and Liu, 1990; Ma and An, 1994; Liu
and Zhang et al., 2005) including potential
alphavirus transmission in Xinjiang (Li and
Liang et al., 1992; Li and Karabatsos et al.,
1995). Therefore, it is of critical importance
to determine the species composition and
seasonal abundance of floodwater mosquitoes in this region. Furthermore, we wanted
to determine if commercial mosquito traps
could be used in residential areas to reduce
local floodwater mosquito populations in
Beiwan, Xinjiang.
II. Materials And Methods
Key Words: Mosquito Magnet® Liberty Plus,
surveillance, Aedes vexans
Study area. The study area is a residential
area enclosed by a dike in the wilderness of
Beiwan, Xinjiang (western China) on the
northern side of the Irtysh River (Figure
1). In this area, there is plentiful vegetation
coverage with shrubs, woods, and reeds. The
northern part of the trapping site is a semidesert area covered with scanty vegetation
on dry sand. The human residential area is
I. INTRODUCTION
Beiwan is located near the Chinese border with Kazakstan and is situated along the
flood lands of the Irtysh River, Xinjiang in
western China. River water levels rise an39
40
Figure 1. Diagrammatic sketch of the study area and trap sites.
located in the transition belt between these
two different landscapes.
Sampling locations. A total of six trapping
sites were selected for the study. Three trapping sites were located along the outside of
the dike in the residential area and were;
riverside (A site), semi-desert (B site), and
a middle zone (C site) between sites A and
B. Three trapping sites inside the residential
area were located along the dike, riverside
(D site), semi-desert (E site), and a middle
zone (F site) between sites D and E.
Mosquito Magnet® Liberty Plus traps
(MMLP) baited with octenol was provided
by the former American Biophysics Corporation, USA. In each trapping site, the same
MMLP was operated in the field for two days.
Mosquitoes in each trap were collected every eight hours (a total of 6 collections) and
brought back to the laboratory for species
identification and total counts. Collections
were made during the peak flood season
(July and August).
Statistical analysis. The proportion of each
species (Pi) in MMLP collections was calculated as Pi (%) = (Ni*100)/N, where N is total
number of mosquitoes and Ni the number of
species “i”. Chi square test was performed to
compare the Pi of different mosquito species
between sites and species (P = 0.05)
III. Results
A total 251,475 mosquitoes representing
six species was collected from MMLPs and
included Aedes vexans (Meigen), Ae. cyprius
Ludl., Anopheles messeae Falleroni, Coquillettidia richiardii (Ficalbi), Culex modestus Ficalbi , and Cx. pipiens pipiens L. (Table 1). Chi
square test results revealed that a significant
Table 1 Composition of mosquitoes trapped with the
Mosquito Magnet Liberty Plus traps in Beiwan, China.
Species
Pi(%)
QTY
Aedes vexans
Ae. cyprius
Anopheles messeae
Coquillettida richiardii
Culex modestus
Cx. pipiens pipiens
Total
97.14
1.30
0.53
0.80
0.09
0.13
100.00
244,279
3,280
1,342
2,007
229
338
251,475
Pi = proportion of each species; QTY = number of mosquitoes
χ2 = 254902, p<0.001
Liu et al.: Mosquito Species Composition and Impact of Trapping Sites on Floodwater Mosquitoes
statistical difference existed between the
proportion of each species (χ2 = 254902,
p < 0.001) with Ae. vexans as the dominant
species (97.14% of total catch). The other
5 species made up about 2.86% of the total
collection.
The proportionth of Ae. vexans, An.
messeae,Cx. modestus, and Cx. pipiens pipiens
caught from the inside human residential
area were higher than those caught outside
(Table 2). The proportion of Ae. cyprius and
Cq. richiardii collected from inside human
the residential area were lower than those
caught from outside the area. Also there was
a significant difference between the proportion of mosquito species outside and inside
the residential area (χ2 = 16.486, p < 0.01).
There was a significant difference in the
proportion of each species between trapping sites inside the residential area (χ2 =
392.86, p < 0.001) (Table 3). Culex pipiens
pipiens from the D site inside the residential
area were higher than the other areas. However, the proportion of Ae. vexans from the
D site area was lower than those collected
from the F site and E site areas. Also, similar
proportions of Ae. vexans, Ae. cyprius, and An.
messeae were collected from the F site and E
site areas. No Cx. pipiens pipiens was collected
from all three sites.
There was a significant difference in
the proportion of each species between the
three trapping sites in the outside residential area (χ2 = 6201.936, p < 0.001) (Table 4).
Aedes vexans and An. messeae collected from
the C site were lower than those collected
from sites A and B; however, the proportion
of Ae. vexans and An. messeae were similar
41
between those 2 latter collection sites. The
proportions of Cx. modestus and Cx. pipiens
pipiens collected from the C site area were
higher than those from the A site and B site.
IV. Discussion
Aedes vexans is a major nuisance species
in northern China (Liang, 1997). Moreover,
Japanese B encephalomyelitis virus has been
isolated from this species in China (Li and Liang et al., 1992; Zhang and Shi et al., 1999).
Our study showed that Ae. vexans still was the
major species and represented 97% of the
total MMLP collections. Because this species
is one of the more proliferate floodwater
mosquitoes, it poses a great health threat to
residents and livestock in this area. Although
few numbers of An. messeae were collected in
MMLPs, this species is a suspected vector of
malaria (Tong and Li et al., 1996; Lu, 1999).
Additionally, Cx. modestus and Cx. pipiens pipiens were collected from traps in low abundance and are suspected vectors of West Nile
virus (Hayes, 1998).
The proportion of Ae. vexans and An. messeae in the riverside were the same as in the
semi-desert environments, which also was
higher than those in the transition zone between these two types of environments. The
species of mosquitoes. Ae. vexans flourishes
in flood environments (O’Malley, 1990). The
flood environment in Beiwan accounted for
the predominance and high proportion of
Ae. vexans in the local mosquito community.
We noticed that density of Cq. richiardii, in
traps from one of our sites, decreased with
increasing distance from the larval habitat as
Table 2 Composition of mosquitoes trapped with the Mosquito Magnet Liberty Plus inside and outside a residential
area in Beiwan, China.
Within residential areas
Outside residential areas
Species
Pi(%)
QTY
Pi(%)
Aedes vexans
Ae. cyprius
Anopheles messeae
Coquillettida richiardii
Culex modestus
Cx. pipiens pipiens
97.83
1.26
0.56
0.01
0.16
0.18
76,190
983
438
4
121
144
96.83
1.32
0.52
1.15
0.06
0.11
Pi = proportion of each species; QTY = number of mosquitoes
χ2 = 16.486, p < 0.01
QTY
168,089
2,297
904
2,003
108
194
42
Table 3. Proportions of mosquito species at different sample sites from Mosquito Magnet Liberty Plus traps in residential areas close to the wilderness, Beiwan, China.
D site
Species
Ae. vexans
Ae. cyprius
An. messeae
Cq. richiardii
Cx. modestus
Cx. pipiens pipiens
F site
E site
Pi(%)
QTY
Pi(%)
QTY
Pi(%)
QTY
93.99
1.71
2.46
0.00
0.88
0.96
9,014
164
236
0
84
92
98.40
1.07
0.30
0.02
0.01
0.21
24,557
267
74
4
3
52
98.35
1.27
0.30
0.00
0.08
0.00
42,619
552
128
0
34
0
Note: Pi = proportion of each species; QTY = number of mosquitoes
D site, the riverside site in residential area;
F site, the semi-desert one in residential area;
E site, is located between F site and D site.
χ2 = 392.86, p < 0.001
observed by other authors (Anno and Takagi et al., 2000 and Leonardo and Rivera et
al., 2005).The larvae of Cq. richiardii can only
be found in reeds from river environments
in China (Lu, 1999). This may explain why
the proportion of Cq. richiardii decreased
with an increase in trapping distance from
the river.
The proportion of An. messeae collected
from inside and outside the human residential area were not different. This may
be caused by the Anopheles mosquitoes. The
proportion of Cx. modestus and Cx. pipiens
pipiens were higher in the human residential
area than outside, but the proportion of Ae.
cyprius and Cq. richiardii in the outside areas
were higher than those in the residential
area. The species variation from different
trapping sites and environments obtained
from our study were similar with other reports from different environments (Ciolpan
and Nicolescu et al., 1990). The trapping
sites with different amounts of vegetation
were significantly influencing the collection
of species and amount of floodwater mosquitoes in the study area.
Our study site, located in the transition
zone between the river and semi-desert environments, undoubtedly influenced which
mosquito species were present and their
relative abundance. The proportion of Ae.
cyprius, An. messeae, Cx. modestus, and Cx.
pipiens pipiens at the trapping sites near the
river were higher than those at the trapping
sites in the center of the human residential
area and near semi-desert land. However, in
the semi-desert region side, the proportion
of Ae. vexans, Ae. cyprius, An. messeae, and Cq.
richiardii at the sites located in the residential
area center were the same as those sites being close to the semi-desert region. There is
plentiful vegetation (such as: reeds, weeds,
Table 4. Proportions of mosquito species at different sample sites from Mosquito Magnet Liberty Plus traps located
in different wilderness areas outside residential areas, Beiwan, China.
A site
C site
B site
Species
Pi(%)
QTY
Pi(%)
QTY
Pi(%)
QTY
Ae. vexans
Ae. cyprius
An. messeae
Cq. richiardii
Cx. modestus
Cx. pipiens pipiens
96.65
0.56
0.56
2.18
0.00
0.04
87,880
512
512
1,984
4
32
96.36
2.76
0.36
0.05
0.23
0.25
34,076
977
128
16
80
88
97.52
1.71
0.56
0.01
0.05
0.16
46,133
808
264
3
24
74
Note: A site, the riverside outside of residential area;
B site, the semi-desert part site outside residential area;
C site, is located between A site and B site.
χ2 = 6201.936, p < 0.001
Liu et al.: Mosquito Species Composition and Impact of Trapping Sites on Floodwater Mosquitoes
shrubs, pygmy trees and some arbors.) which
might provide good resources and resting
sites for adult mosquitoes.
Our results tell us that surveillance of
the mosquito density and species composition is important to present good protection for people and livestock. Human-baited
bed nets had previously been used for adult
mosquito surveillance in this area (Liu and
Zhang et al., 2005). Our study found that Ae.
vexans was the major species in the area (Pi
of this species was 99.5%) while Ae. cyprius
and An. messeae were also collected. In addition, the Mosquito Magnet® Traps with octenol collected a great number of floodwater mosquitoes and this method may reduce
the nuisance problem in Beiwan where an
extremely high density of mosquitoes occurs
during the flood seasons. Mosquito Magnet
Liberty Plus traps are an efficient surveillance tool for mosquito population and
could be considered for use as control measures to reduce the mosquito population in
the residential area (Burkett and Lee et al.,
2002; Kline, 2006).
V. References cited
Anno, S. and M. Takagi, et al. (2000). Analysis of relationship between Anopheles subpictus larval densities
and environmental parameters using Remote Sensing (RS), a Global Positioning System (GPS) and
a Geographic Information System (GIS). Kobe J.
Med. Sci. 46:231-243.
Burkett, D. and W. Lee, et al. (2002). “Late season commercial mosquito trap and host seeking activity eval-
43
uation against mosquitoes in malaria’s area of the
Republic of Korea.” Korean J. Parasitol (40): 45-54.
Ciolpan, O. and G. Nicolescu, et al. (1990). Investigatii
asupra faunei de tîntari (Diptera: Culicidae) din
padurea Cernica, potential vectori pentru arbovirusuri. Analele Banatului, Stiintele Naturii 2:356-360.
Hayes, C. (1998). West Nile fever. In: The Arboviruses: Epidemiology and Ecology. T.P. Monath (ed.) CRC Press,
Boca Raton, Florida,.
Kline, D. L. (2006). Traps and trapping techniques for
adult mosquito control. J. Am. Mosq. Control Assoc.
22:490-6.
Leonardo, L. and P. Rivera, et al. (2005). A study of the
environmental determinants of malaria and schistosomiasis in the Philippines using remote sensing
and geographic information systems. Parassitol.
47:105-114.
Li, Q. and G. Liang, et al. (1992). Survey of arboviruses
in Xinjiang. Chin J. Exp. Clin. Virol. 6:247.
Li, Q. and N. Karabatsos, et al. (1995). Further identification of some arboviruses isolated in Xinjiang,
China in recent years. Chin. J. Exp. Clin. Virol.
9:367-370.
Liang, G. (1997). The arbovirus research situation in
China. Chin. J. Zool. 13:61-63.
Liu, B. and G. Zhang, et al. (2005). Investigation of the
trends of growth and decay of Aedes vexans in the
daytime and at night in the uninhabited region of
Beiwan in Xinjiang province. Chin. J. Hyg. Insecticides and Equipment 11:103-105.
Lu, B. (1999). Progress of studies of mosquito vectors in
China. Chin. J. Vect. Biol. Control 10:161-165.
Lu, B. and Z. Liu (1990). Medical Entomology. Beijing,
China, Beijing Science Publishing House.
Ma, D. and J. An (1994). Survey of mosquito composition and ecology in the A’letai district of Xinjiang.
Chin. J. Vect. Biol. Control 5:48-49.
O’Malley, C. M. (1990). Aedes vexans (Meigen): An old
foe. Proc. NJ Mosq. Control Assoc. 1:90-95.
Tong, S. and B. Li, et al. (1996). Study on the surveillance of vivax malaria in the Yili Area. Xinjiang. Endem. Dis. Bull. 11:72-74.
Zhang, H. and H. Shi, et al. (1999). Isolation of Japanese encephalitis virus from four species of Aedes
mosquito in Yunnan Province. Virol. Sin. 14:32-35.
LUNAR PHASE IMPACT ON COQUILLETTIDIA
PERTURBANS AND CULEX ERRATICUS HOST SEEKING
IN NORTHERN FLORIDA
YONGXING JIANG
Mosquito Control Section
Department of Public Works, City of Gainesville
405 NW 39th Ave, Gainesville, FL 32609
ABSTRACT. The host seeking periodicity
of Coquillettidia perturbans and Culex erraticus
(two important mosquito bridge vectors of
eastern equine encephalitis virus in northern
Florida) were determined by using programmable collection bottle rotary traps. Results
showed that both species host seeking activity peaked 1 hour after sunset while Cx. erraticus continued that level of activity for an
additional two hours. On full moon nights,
both species displayed peak host seeking
activity 1-2 hours longer than that observed
during non-full moon nights. High relative
humidity significantly increased Cx. erraticus
abundance in CDC light traps, as well as host
seeking activity but not Cq. perturbans.
atmospheric moisture, wind, moonlight, and
mosquito physiological age may influence
the host seeking behavior of mosquitoes on
any given night. Therefore, it is important to
have some understanding of the prevailing
meteorological conditions and how these
conditions modify the behavior of different
mosquito species, and consequently the efficacy of controlling those species. During
2008-2009, CDC light traps were placed
in four northern Florida counties (Leon,
Madison, Holmes and Washington) where
positive eastern equine encephalitis (EEEV)
equine or sentinel chicken sera were reported to determine what mosquito species were
in those areas. Coquillettidia perturbans (Walker) and Culex erraticus (Dyar and Knab) were
the dominant species in all collections and
may have been involved in virus transmission. Both species have been incriminated
as competent EEEV bridge vectors in North
America (Howitt et al. 1949, Boromisa et al.
1987, Cupp et al. 2003, 2004).
McNeel (1932), in an observational
study, found that Cq. perturbans host seeking
activity was greatest at dusk. Snow and Pickard (1957) presented data that indicated
Cq. perturbans host seeking activity peaked
between 1930-2000 h. However, results from
these earlier observational studies were not
in agreement with a recent study conducted
by Bosak et al. (2001). These authors revealed that significantly more Cq. perturbans
host seeking activity occurred during the
night period (between 2200-0100 h) compared with early evening or morning hours.
Because of the inconsistencies between the
previous studies there is a need to re-examine the host seeking pattern of this species
KEY WORDS. Coquillettidia perturbans, Culex
erraticus,host seeking pattern, lunar phase,
humidity
I. INTRODUCTION
Knowledge of temporal host seeking
patterns of mosquitoes is important for understanding mosquito-host contact, as mosquitoes will only bite hosts that are available
during the period of their host seeking activity (Williams 2005). The time of host seeking by mosquitoes delineates periods when
females are active and the risk of pathogen
transmission greatest for vector species (Reisen et al. 1997). Therefore, determination
of the host seeking time (or biting rhythms)
exhibited by vector and nuisance mosquito
species is essential to decide when adulticides can be most effectively applied. Obviously, climatic factors such as temperature,
44
Jiang: Lunar Phase Impact on Host Seeking in Northern Florida
in northern Florida especially in areas where
EEEV is prevalent.
As stated earlier, Cx. erraticus was also one
of the more abundant species collected in
light traps from EEEV sites and may be an
important bridge vector in the area. To this
author’s knowledge, host seeking periodicity of this species has not been reported in
the scientific literature. Therefore, the aim
of this study was to determine the temporal
host seeking activity for Cq. perturbans and
Cx. erraticus and identify some of the meteorological factors that may affect timing
of this behavior. Obtaining this information
will provide data-driven guidance on when
adulticides should be most effectively applied.
Study site. Field studies were conducted
at Leon County’s solid waste management
center,7550 Apalachee Parkway, Tallahassee,
Florida (30.25.362 N, 084.08.956 W). This location is one of Leon County Mosquito Control’s permanent sentinel chicken sites that
is used to monitor mosquito-borne diseases
such as EEEV and West Nile virus in Leon
County. This site consisted of a large fresh
water swamp (6.1 ha) covered with cattails
(Typha spp.), arrowhead weeds (Sagittaria
longiloba), pickerelweed (Pontederia cordata),
a large forested wood lot, and office buildings. Seropositive EEEV chickens have been
reported from this site for the last five years
(http://www.floridahealth.gov/diseasesand-conditions/mosquito-borne-diseases/
surveillance.html).
Mosquito collection and processing. Programmable collection bottle rotator traps
(CBR) (Model 1512, John W Hock Company, Gainesville, FL) were used to determine
temporal host seeking periodicity of Cq. perturbans and Cx. erraticus. The CBR is a device
that segregates collections into distinct time
periods by a programmable timer to which
any mechanical mosquito trap, such as a
CDC light trap, is attached. In this study, a
standard CDC light trap was mounted onto
the screen holder located on the top of the
CBR unit and secured with thumb screws. A
45
central stainless steel rod extended from underneath the rotator trap and inserted into
a tripod lawn sprinkler stand that supported
the unit and CDC trap 1.5 meter above the
ground. The first collection started at 1943
h (1 hour before sunset) and ended after
the collection at 0639h (sunrise). Because
each rotator trap only contained 8 bottles,
two traps were used to achieve a total of 10
hourly collections. In order to synchronize
sunset/sunrise collections, the programmable timer was adjusted accordingly to reflect
daily time changes. A separated CDC light
trap was set up about 50 m apart from the
CBR traps at the edge of the swamp in order
to compare species composition between
the two collecting methods. CDC and CBR
traps were baited with 5 lbs of dry ice contained in a one gallon cooler jug. A small
hole was drilled at the bottom of jug so the
CO2 could be delivered to the traps directly.
Traps were operated for nine nights during
July 1 through August 8, 2009. Mosquito
collections were transported to the laboratory, anesthetized, identified to species, and
counted. The number of Cq. perturbans and
Cx. erraticus trapped in each hourly bottle
collection were summed and an average
number of mosquitoes per hour calculated
for each species.
Meteorological data. On each collection
day, temperature, relative humidity, and
wind speed were obtained from a www.wunderground.com weather station located
about 2 miles from the study site. Moon
phase data were obtained from www.moonconnection.com. For this study, the night of
a full moon and the night following a full
noon were considered as “full moon nights”.
(Criterion for this determination was based
on the moon phase calendar where the
night following a full noon is closer to a full
moon than the night before a full moon).
A total of four full moon night collections
were made in this study, (two nights in July
and two nights in August). The remaining
sample nights were considered as non-full
moon nights.
Statistical analysis Mosquito collections
were summarized on the basis of the number of females caught per trap per hour. To
46
determine if temperature, relative humidity,
and/or wind speed impacted the host seeking behavior of Cq. perturbans and Cx. erraticus, one way analysis of variance was used to
analyze these relationships using STATISTICA software (StatSoft Inc. 2001) (P < 0.05).
Hourly mosquito counts were log-transformed
[log (n + 1)] to normalize the distribution and
relative abundance prior to analysis.
III. RESULTS
Mosquito collection. Mosquito collection
started from July 1 and ended on August 8,
2009. During this 10 week collection period,
a total of 9 nightly collections were made. A
total of 2,984 female mosquitoes representing 16 species were collected by the CDC light
trap, and 1,705 mosquitoes representing 12
species were collected by the CBR traps. Aedes albopictus (Skuse), Ae. infirmatus (Dyar and
Knab), Ae. triseriatus (Say), Ae. vexans (Meigen),
Anopheles crucians Wiedemann, Cq. perturbans,
Cx.erraticus, Cx. nigripalpus Theobald, Psorophora columbiae (Dyar and Knab) and Ps. ciliata (Fabricius) were collected in both types of
traps. The composition of Cq. perturbans and
Cx. erraticus was 40.7%, 48.4% in CDC traps
and 37.1%, 57.1% in CBR traps, respectively.
Compared with the CBR, more mosquito spe-
cies (15 species from CDC traps vs. 12 species
from CBR traps) and higher numbers of mosquitoes (2,984 female mosquitoes from CDC
trap collection vs. 1,705 female mosquitoes
from CBR collection) were collected by the
CDC traps. In terms of percentage, Cq. perturbans collections from CDC traps were slightly
greater than CBR traps (40.7% vs. 37.1%); in
contrast, a higher percentage of Cx. erraticus
was collected in CBR traps than CDC trap
(57.1% vs. 48.4%), respectively.
Host seeking periodicity. Figure 1 shows the
temporal host seeking patterns of Cq. perturbans and Cx. erraticus. Overall, Cq. perturbans
host seeking activity started to rise at sunset
(SS) then peaked 1 hour after sunset (SS+1),
followed by a gradually reduction in activity through the night until one hour before
sunrise.
Culex erraticus peaked at sunset and sustained a higher level of host seeking activity
for an additional two hours (SS+2), followed
by a sharp reduction in host seeking activity 3
hr after sunset (SS+3). Activity then gradually
slowed down throughout the night. Host seeking activity was lowest 3 hours before sunrise.
Host seeking periodicity at full-moon and
non-full nights. Figure 2 shows the temporal
pattern of host-seeking activity for Cq. perturbans and Cx. erraticus as influenced by lu-
Figure 1. Overall host seeking patterns of Coquillettida perturbans andCulex erraticus. SS, sunset time; SR, sunrise
time. *indicates that significantly (P<0.05) differences between the number of mosquitoes collected at SS+2 and
SS+3 hr.
47
Figure 2. Comparison of host seeking patterns of Coquillettida perturbans (Cq.) and Culex erraticus (Cx.) under different moon phase nights. SS, sunset time; SR, sunrise time. * *indicates significant differences (P<0.01) between
the number of mosquitoes at the non-full and full moon nights.
nar phases. Both species delayed their peak
host seeking activity at least 1 hour on full
moon nights than non-full moon nights accompanied by an increase in the number of
mosquitoes trapped during this extended
activity period. In terms of Cq. perturbans, its
peak activity was one hour later at full moon
nights (SS+2) than that of non-full moon
nights (SS+1).The number of mosquitoes
collected at SS time and SS+1 were not much
different (Figure 2), however, the numbers
collected at SS+3, SS+4, SS+5 and SS+6 were
much higher (no statistical significantly differences) at the full-noon nights than that of
the non-full moon nights. Furthermore, Cq.
perturbans extended its activity by two hours
on full-moon nights (SS to SS+3) compared
with non-full moon nights (i.e. SS to SS+1).
Culex erraticus, on the other hand, exhibited a two hour delay in peak host seeking
activity on full-moon nights (SS+2) compared
with non-full moon nights (SS) (Figure 2).
Significantly more mosquitoes (P<0.05) were
trapped at SS+1, SS+2 and SS+4 on full moon
nights than non-full moon nights.
Host seeking periodicity under different meteorological conditions. Host seeking activity
of Cq. perturbans was not significantly influenced by changes of temperature, relative
humidity, or wind speed (P<0.362; P<0.463;
P<0.088; respectively). Temperature and
wind speed did not significantly effect Cx. erraticus (P<0.48; P<0.263; respectively). However, significant (P<0.01) higher numbers of
Cx. erraticus were trapped at higher humidity
(85-95%) than low humidity and indicated
that this meteorological factor altered Cx. erraticus host seeking behavior.
IV. DISCUSSION
Information regarding the precise timing of host seeking by Cq. perturbans and Cx.
erraticus is essential for the optimal timing
of adulticide missions. This study clearly indicated that Cq. perturbans host seeking activity started at sunset and sustained a peak
level of activity for at least two hours. This
is in contrast to that reported by Snow and
Pickard (1957) where peak host seeking occurred between 1930-2000 h and Bosak et al.
(1987) from 2200 to 0100 h. It is likely that
mosquitoes react to light intensity crepuscular changes (sunset, sunrise) to initiate their
flight and host seeking rather than actual
“clock time” because the periods of sunrise/
sunset change over the seasons. Moreover,
temporal host seeking by Cq. perturbans may
vary in different geographic regions and the
use of crepuscular light intensity changes
48
may be a more appropriate indicator of host
seeking flight behavior.
Although Cx. erraticus dispersal behavior
(Estep et al. 2010), host feeding preference
(Oliveira et al. 2011; Burkett-Candena et al.
2012; Mendenhall et al. 2012) and general
bionomics (Robertson et al. 1993) have
been extensively studied, temporal host
feeding periodicity has not been reported.
This is the first report to describe accurately
the timing of host seeking behavior of Cx.
erraticus in northern Florida, as well as the
influence of meteorological conditions,
such as moon phase and relative humidity,
on host seeking behavior.
Coquillettidia perturbans and Cx. erraticus
are considered crepuscular-nocturnal species; however, their host seeking patterns
have similarity and differences. For example, both species consistently commenced
host seeking at sunset, either continued or
slightly increased its host seeking activity for
the next two hours. However, Cq. perturbans
reduced its host seeking activity gradually at
SS+2, whereas, Cx. erraticus host seeking activity abruptly dropped at SS+2.
Many mosquito species have their greatest
period of flight activity during twilight (Mitchell, 1982; Nelson and Spadoni, 1972; Reisen et
al. 1983). Moonlight can simulate twilight conditions by increasing nocturnal illumination.
This study also illustrates that Cq. perturbans
and Cx. erraticus significantly their increased
activity and an extended activity period under the full moon phase (Figure 2), which is
consistent with early studies. Antonipulle et
al. (1958) studied the host seeking behavior
of Mansonia uniformis (Theobald) using cattlebaited traps. Analysis of the catches by hour
showed that during each lunar phase, the
highest approach rates occurred during the
periods of full moonlight. Ribbands (1946)
also found that the number of Anopheles funestus Giles entered man-baited huts was significantly correlated with the times of twilight
and of moonlight. Enter rates were low during
moonless periods of the night and highest at
the full moon period. However, exactly the opposite results were reported by Chadee and Tikasingh (1989). Their studies showed that over
70% mosquito Culex caudelli Dyar and Knab
captured in mouse-baited traps were collected
between 2200 and 0400 h. They compared the
collections made during the four lunar quarters and found out that neither the extent of
host seeking activity nor its timing was affected
by moon phase. The impact of lunar phase on
the host seeking patterns of mosquitoes may
vary by species, physiological age, and meteorological conditions (Provost 1959).
It is important to understand ambient
meteorological conditions as they affect
the behavior of various mosquito species.
Temperature, relative humidity, and wind
speed affect mosquito behavioral periodicities as permissive factors. This study showed
that there was no correlation between host
seeking periodicity and the meteorological
parameters recorded during collections for
Cq. perturbans. However, relative humidity
had a significantly (P < 0.01) positive impact on Cx. erraticus host seeking patterns.
A statistically significant increase in the
number of Cx. erraticus collected during
periods of high relative humidity (85-95%)
was reported in this study. Day and Curtis
(1989) reported a significant positive correlation between blood feeding by Cx. nigripalpus and relative humidity. Dow and
Gerrish (1970) also reported a positive correlation between host seeking of Cx. nigripalpus in baited traps and humidity with a
4.8% increase in mosquito catches for each
1% increase of relative humidity. Evidently,
the meteorological factors such as humidity
had influence on Cx. erraticus host seeking
behavior but not Cq. perturbans. The impact
was more species-specific, as in the case of
Cx. erraticus in this study.
The precise timing of an adulticide
spray application designed to intersect
with maximum adult mosquito flight behavior is critical for effective mosquito
control. This study reveals the accurate
timing of the crepuscular/nocturnal flight
patterns of Cq. perturbans and Cx. erraticus,
two potential EEEV bridge vectors in North
Florida. Results from this study could be
providing the optimal time for controlling
the two species of mosquitoes and reducing the risk of EEE virus transmission in
North Florida.
V. ACKNOWLEDGEMENTS
The author would like to thank Leon
County Mosquito Control for helping to
identify mosquito trap locations and agreeing to use their sentinel chicken sites for this
study.
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Williams, C. R. 2005. Timing of host-seeking behavior of
the mosquitoes Anopheles annulipes sensu lato Walker
and Coquillettidia linealis (Skuse) (Diptera: Culicidae) in the Murray River Valley, South Australia.
Aust. J. Entomol. 44:110-112.
FIELD EVALUATION OF THREE COMMERCIAL MOSQUITO
TRAPS AND FIVE ATTRACTANTS IN NORTHEASTERN
FLORIDA
ALI FULCHER, RACHEL SHIRLEY, MICHAEL L. SMITH, JODI M. SCOTT,
AND RUI-DE XUE
500 Old Beach Road, St. Augustine, FL, 32080
ABSTRACT. Three commercial mosquito
traps and five attractants were evaluated in
northeastern Florida. The combinations
of traps and attractants were BG Sentinel (BGS) + BG Lure, Mosquito Magnet X
(MMX) + CO2, Mos-Hole + naphtha, MMX
+ octenol, Mos-Hole + BG Lure, BGS + octenol, and BGS + Lurex3. The MMX trap
paired with CO2 collected the greatest number of mosquitoes and the MMX trap paired
with octenol trapped the least amount compared with the rest of the trap/attractant
combinations.
acid, octenol, ammonia, or fatty acids) collect a variety of mosquito species (Kline et
al. 1991, 2006, Ritchie et al. 2006, Farajollahi et al. 2009, Hoel et al. 2009, Cilek et al.
2011, Xue and Smith 2013). Recently, Xue
et al. (2015) found the novel Mos-HoleTM
trap, baited with naphtha, was an effective
tool for surveillance of anthropophilic mosquitoes when compared with BioGents SentinelTM (BGS) and MMX traps. In this paper
we report on additional field evaluations we
conducted regarding the attractiveness of
BGS, MMX, and Mos-Hole traps baited with
either CO2, naphtha, octenol, lactic acid, or a
commercial BG Lure™ to collect adult mosquitoes in St. Johns County, Florida.
Key Words. BG Sentinel trap, Mos-Hole trap,
Mosquito Magnet X trap, BG lure, octenol
I. INTRODUCTION
Field trials were conducted in St. Johns
County during June and July 2013. The location of the study site was off of County Road
214 (29.87.13N, 81.36.98W). This low-lying
hardwood habitat contained several hundred used tires (16 m W × 133 m L × 4 m
H) surrounded by agricultural land with
shallow ditches. BGS (white cover version)
(BioGents, Regensburg, Germany), MosHole (Lnd E-TND Co. Ltd., Hanam City,
Gyeongi-Do, Republic of Korea), and MMX
(Woodstream Corportation, Litiz, PA) traps
were used in this study and powered by 12v
batteries. The Mos-Hole trap was a dark blue
unit placed on the ground surface similar to
how BGS traps are positioned.
The following five attractants were tested: BG Lure (ammonia, lactic acid, and fatty
acids, BioGents, Regensburg, Germany), liquid naphtha (Lnd E-TND Co. Ltd., Hanam
Mosquitoes are highly dependent on
their olfactory capabilities to host seek (Anton and Rospars 2004). Semiochemicals
(e.g. octenol and CO2) and physical characteristics (such as convection currents and
warmth) can signal the presence of a host.
Historically, mosquito traps have been baited with a variety of attractant substances that
take advantage of the host seeking instinct
by mimicking a blood meal source or chemicals exuded by animal hosts. Generally, from
the perspective of the Anastasia Mosquito
Control District (AMCD), there are no traps
or attractants that are universally attractive
to all 43 species of mosquitoes found within
its jurisdiction.
A number of studies have reported that
Mosquito Magnet® X traps (MMX) with
CO2 and other attractants (including lactic
50
Fulcher et al.: Field Evaluation of Three Commercial Mosquito Traps and Five Attractants City, Gyeongi-Do, Republic of Korea), CO2
(dry ice block from local supplier), octenol
(BioSensory, Willimantic, Connecticut) and
Lurex3 (ammonia and lactic acid, Woodstream Corportation, Litiz, PA). A total of
seven trap/attractant combinations were
evaluated (Table 1). Liquid naptha is a flammable liquid hydrocarbon that when combined with air, produces heat and CO2 that
was specifically designed for the Mos-Hole
trap (Xue et al. 2015). The liquid was contained in a polyethylene bottle and placed
on the bottom surface inside the trap. Also
a mesh pouch was sewn into the Mos-Hole
trap to mimic the pouch of the BGS, so that
both traps were able to contain attractants.
The MMX trap was suspended from a shepherd’s hook so that the trap opening was
approximately 0.5m from the ground surface. Carbon dioxide was dispensed to this
trap from a pressurized gas cylinder through
5 mm silicon tubing and controlled using
a flow-meter. BG Sentinel and MMX traps
plus naphtha and MMX plus Lurex3 were
not evaluated in this study, because they had
been evaluated earlier by Xue et al. (2015).
Traps were set up in the shade in a linear
fashion, 10 m apart, and >15m from the
closest the tire pile. Traps were rotated and
rebaited daily after collection. After each 24hour trapping period, the labeled collection
bags were transported to the AMCD laboratory, frozen to -20° C, and identified to species using the taxonomic keys of Darsie and
Ward (2004).
Statistical analysis. Mean mosquito abundance from each trap/lure combination was
separately pooled by genera (Aedes, Culex,
and Psorophora) and subjected to one-way
51
ANOVA using JMP® ver. 11.1 software (SAS
Institute, Inc. 2013) after log transformation
of trap count data. Differences were considered significant at P ≤ 0.05. A Tukey-Kramer
HSD multiple mean test was used to establish genera differences within trap/lure
combinations.
A total of 1,207 mosquitoes from 15 species were collected during the study period
representing 8 genera. Overall mosquito
composition and abundance in collections
were as follows: Aedes albopictus (Skuse)
(27.9%), Culex nigripalpus Theobald (26.6%),
Ae. infirmatus (Dyar and Knab) (26.5%), Culiseta melanura (Coquillett) (4.0%), Psorophora
ferox (von Humboldt) (3.2%), Ps. columbiae
(Dyar and Knab) (3.0%), Ae. atlanticus (Dyar
and Knab) (1.8%), Anopheles quadrimaculatus Say (1.6%), An. crucians Wiedemann
(1.6%), Ae. triseriatus (Say) (0.8%) Ae. fulvus
pallens (Ross) (0.2%), Ps. howardii Coquillett
(0.1%), Toxorhynchites rutilus (Coquillett)
(0.1%) and Cx. quinquefasciatus Say (0.1%).
Of the total mosquitoes collected 0.7% were
unable to be identified due to damage.
There was no statistical difference when
traps were compared with total mosquitoes
collected (F = 2.984, P = 0.055) as well as
attractants and mosquito totals (F = 1.778,
P = 0.138). Overall, there was a significant
difference in mosquito abundance between
the seven trap/attractant combinations (F =
6.240, P = 0.004). The MMX trap paired with
CO2 collected the greatest number of mosquitoes followed by BGS+Lurex3, BGS+BG
Lure, Mos-Hole+naphtha, Mos-Hole+BG
Table 1. Mean number of adult mosquitoes from three genera collected by BG Sentinel (BGS), Mos-Hole, and
Mosquito Magnet X (MMX) traps and five attractants from St. Johns County FL1
Trap
Culex
Aedes
Psorophora
MMX + CO2
BGS + Lurex3
BGS + octenol
BG + BG Lure
Mos-Hole + BG Lure
Mos-Hole + naphtha
MMX + octenol
84.7a
4.0c
2.0bc
2.3c
2.0c
8.3c
1.0c
39.7b
23.0bc
12.0c
6.6c
11.7c
10.9c
NA
7.3c
6.0c
1.5c
2.0c
2.0c
1.5c
NA
Means followed by the same letters in a row are not significantly different (P >0.05).
1
52
Lure, BGS+octenol, while the MMX trap
paired with octenol trapped the least amount
of mosquitoes.
No significant difference existed between
each of the 15 mosquito species trapped by
the 7 trap/attractant combinations. However,
there were differences (F = 3.589, P = 0.0001)
between abundance of the three genera within trap/attractant combination (Table 1).
MMX traps plus CO2 collected significantly
more Aedes and Culex mosquitoes than Psorophora. However, trap abundance for the rest
of the trap/attractant combinations were no
different between genera.
Previously, Xue et al. (2015) found that
Mos-Hole traps baited with naphtha were an
effective tool for surveillance and control for
anthropophilic mosquitoes compared with
BGS+BG Lure and MMX traps baited with
Lurex3. That study did not find differences
in the number of individuals, per genera,
from BGS traps baited with naphtha. Xue
et al. (2015) also reported that Mos-Hole
traps baited with BG Lure collected greater
numbers of Aedes mosquitoes than BGS traps
baited with naphtha. We noticed the same
trend for Mos-Hole traps baited with either
BG Lure or naphtha in our study (Table 1).
We also found that the MMX trap plus
octenol was least effective. This contradicted
the results from Xue et al (2010) who reported that the same trap/attractant combination
collected more floodwater mosquitoes than
similar traps baited with Lurex3 and a CO2
sachet. Xue et al. (2010) also found Lurex3
and octenol were effective attractants when
combined with MMX traps or the Mosquito
Magnet® Pro, in order “to improve the collection and possibly the management of floodwater mosquitoes”. Although the BGS trap
plus Lurex3, or BG Lure, did not collect as
many total mosquitoes (as MMX plus CO2)
they captured the second highest number of
Psorophora. Our data suggests that the BGS
trap/Lurex3 combination targeted floodwater species more than expected because the
attractant sachet (which contained components of human odor) should have drawn
more anthropophilic mosquitoes to the trap.
Out of all seven different trap and attractant combinations tested, the MMX trap
paired with CO2 was the overall best combination. Hoel et al. (2007) reported similar results
where un-baited Mosquito Magnet Pro traps
caught the largest number of floodwater species. Moreover, these authors also found that
their traps, when baited with octenol and lactic
acid, actually decreased collection abundance.
In studies by Xue et al. (2008) and Zhu et al.
(2014) dry ice baited MMX traps were very effective for collecting floodwater mosquitoes
from multiple field sites
MMX traps remain effective tools for mosquito surveillance because the units intake
entrance, while positioned at the bottom,
takes advantage of the natural tendency of
mosquitoes to fly up. Moreover, MMX traps
are ideal for research purposes when mosquitoes need to remain alive during collection.
The trap’s collection chamber is able to hold
a water-soaked cotton ball for moisture and
nutritional requirements if this is required by
the trapped mosquitoes. The BGS and MosHole traps have fairly small mesh collection
bags. Mosquito specimens in these traps tend
to be easily desiccated and damaged by the
air velocity of their intake fans.
In summary, continuing studies are warranted that evaluate the effectiveness of
traps without attractive substances and/or
combining additional attractants in order to
determine additive or synergistic effects on
trap abundance and species composition.
Comparative studies should consider evaluation with traps that are currently considered
the gold standard for mosquito surveillance
in order to increase the effectiveness of operational programs.
IV. ACKNOWLEDGMENTS
The authors are grateful to Claudia Davidson, Tanjim Hossain, Richard Weaver,
and Kay Gaines for their technical help.
This is a research report only and does not
endorse any of the commercial products involved or mentioned in this report.
IV. REFERENCES CITED
Anton, S. and J. P. Rospars. 2004. Quantitative analysis
of olfactory receptor neuron projections in the an-
Fulcher et al.: Field Evaluation of Three Commercial Mosquito Traps and Five Attractants tennal lobe of the malaria mosquito, Anopheles gambiae. J. Comp. Neurol. 475:315–326.
Cilek, J. E., C. F. Hallmon, and R. Johnson. 2011. Semifield comparison of the BG Lure, nonanal, and 1-octen-3-ol to attract adult mosquitoes in northwestern
Florida. J. Am. Mosq. Control Assoc. 27:393-7.
Darsie, R. F. and R. A. Ward. 2004. Identification and Geographical Distribution of the Mosquitoes of North America,
North of Mexico. University Press of Florida, Gainesville, FL.
Farajollahi, A., B. Kesavaraju, D. C. Price, G. M. Williams, S. P. Healy, R. Gaugler, and M. P. Nelder. 2009.
Field efficacy of BG-Sentinel and industry-standard
traps tor Aedes albopictus (Diptera: Culicidae) and
West Nile virus surveillance. J. Med. Entomol.
46:919–925.
Hoel, D. F., D. L. Kline, and S. Allan. 2007. Evaluation of
carbon dioxide, 1-octen-3-ol, and lactic acid as baits
in Mosquito Magnet Pro traps for Aedes albopictus in
north central Florida. J. Am. Mosq. Control Assoc.
25:47-57.
Hoel, D.F ., D. L. Kline, and S. Allan. 2009. Evaluation
of six mosquito traps for collection of Aedes albopictus
and associated mosquito species in a suburban setting in north central Florida. J. Am. Mosq. Control
Assoc. 25:47-57.
Kline, D. L. 1994. Olfactory attractants for mosquito
surveillance and control: 1-octen-3-ol. J. Am. Mosq.
Control Assoc. 10(2 Pt 2):280-287.
Kline, D. L., D. A. Dame, and M. V. Meisch. 1991.
Evaluation of 1-octen-3-ol and carbon dioxide as
attractants for mosquitoes associated with irrigated
rice fields in Arkansas. J. Am. Mosq. Control Assoc.
7:165-169.
53
Kline, D.L. 2006. Traps and trapping techniques for
22:490–496.
Ritchie, S.A., P. Moore, M. Carruthers, C. Wolliams, B.
Montgomery, P. Foley, S. Ahboo, A.F. Van Den Hurk,
M.D. Lindsay, B. Cooper, N. Beebe, and R.C. Russell.
2006. Discovery of a widespread infestation of Aedes
albopictus in the Torres Strait, Australia. J. Am. Mosq.
SAS Institute, Inc. 2013. JMP®, Version 11.1. Cary, NC.
Xue, R. D., W. A. Qualls, D. L. Kline, and T. Y. Zhao.
2010. Evaluation of Lurex3™, octenol, and CO2 sachets as baits in mosquito magnet pro traps against
flood water mosquitoes. J. Am. Mosq. Control Assoc.
26:244–245.
Xue, R. D. and M. L. Smith. 2013. Field evaluation of
octenol, Lurex3, BG-Lure, and octenol+Lurex3
combination as baits in BG Sentinel traps to collect
Aedes albopictus. Tech. Bull. Fl. Mosq. Control Assoc.
9:21–23.
Xue, R. D., M. L. Smith, H. Yi, and D. L. Kline. 2015.
Field evaluation of a novel Mos-Hole trap and naphtha compared with BG Sentinel trap and Mosquito
Magnet X trap to collect adult mosquitoes. J. Am.
Xue, R. D., M. A. Doyle, and D. L. Kline. 2008. Field evaluation of CDC and Mosquito Magnet X traps baited
with dry ice, CO2 sachet, and octenol against mosquitoes. J. Am. Mosq. Control Assoc. 24:249–252.
Zhu, L., A. Fulcher, T. Hossain, C. Davidson, J.C. Beier,
and R. D. Xue. 2014. Body size, blood feeding activity and fecundity of Psorophora howardii, Psorophora
ciliata and Psorophora ferox (Diptera: Culicidae). J.
FIELD EVALUATION OF MOSQUIRON 0.12CRD AGAINST
CULEX QUINQUEFASCIATUS IN STORM DRAINS,
DOWNTOWN ST. AUGUSTINE, FLORIDA
ALI FULCHER, RUI-DE XUE, JODI M. SCOTT, MICHAEL L. SMITH,
MARCIA K. GAINES, AND JAMES H. R. WEAVER
ABSTRACT. A controlled release formulation of the chitin synthesis inhibitor novaluron (Mosquiron 0.12 CRD) was evaluated
in downtown St. Augustine, FL storm drains
for 8 weeks. Results suggested that the treatments provided effective control of larval
Culex quinquefasciatus in storm drains that
were cleaned of debris prior to treatment.
Synchronizing treatment with municipalities’ drain debris cleaning schedules may improve the effectiveness of this formulation
against mosquito larvae.
Treatment of individual storm drains by
mosquito control district personnel can be
laborious, time consuming, and costly. Many
controlled release larvicide formulations are
expensive. Some sink to the bottom and are
buried in sediments, while others may be
washed away during heavy storm and rainfall
events. Anastasia Mosquito Control District
currently uses Bacillus thuringiensis israelensis (Bti) liquid in downtown St. Augustine
storm drains to achieve successful control.
However, this formulation has a limited
treatment window where application must
occur before fourth instar development because feeding slows down as they approach
later instars. The other challenges of Bti formulations include short residual life and the
adverse impact that organic matter, in the
drain, has on product efficacy.
Past treatment of storm drain habitats
by AMCD in downtown St. Augustine has
focused on thermal fogging and toxic bait
stations to control adult mosquitoes (Müller
et al. 2010, Xue et al. 2012, Xue et al. 2013).
Controlling larval stages in storm drains before they become adults has advantages over
adult control. The ideal profile of a larval
control product is that it would be reasonable
in cost, provide long residual efficacy, and
remain in the application location through
significant rainfall events. With an increase in
storm drains with higher capacities, new larvicide products that provide controlled release
of the active ingredient are critically needed.
Therefore, we report here on the evaluation
of a novel controlled release larvicide formulation, Mosquiron® 0.12 CRD, (novaluron
0.12%AI) against mosquito larvae in downtown storm drains in St. Augustine, Florida.
Key Words. Culex quinquefasciatus, novaluron, insect growth regulator, storm drain
I. INTRODUCTION
In Florida, the number of suburban and
urban storm drains has been steadily increasing due to continued growth associated
with residential and infrastructural development. As the number of storm drains have
increased, their water and debris holding
capacity have also increased as a result of recent National Pollutant Discharge Elimination System (NPDES) regulations (Metzger
2004). As older infrastructure is replaced
with NPDES compliant ones, a subsequent
increase in potential larval production sites
may be inevitable (Su et al. 2003a). An increase of these new storm drains in close
proximity to humans increases the challenges faced by the Anastasia Mosquito Control
District (AMCD).
West Nile virus vectors, like Culex quinquefasciatus Say, thrive in storm drains and will
readily feed on humans (Moulis et al. 2013).
54
Fulcher et al.: Field Evaluation of Mosquiron 0.12Crd Against Culex quinquefasciatus55
Since 2003, various formulations of novaluron have been evaluated against Culex
species in laboratory and field studies (Su
et al. 2003b). Interestingly, application
rates that achieved <80% control appear to
be species dependent (Arredondo-Jiminez
and Valdez-Delgado, 2006). The mode of
action of novaluron is that of a chitin synthesis inhibitor within the class of insect
growth regulators. Early studies found lower concentrations of novaluron primarily
resulted in inhibition emergence of adults,
while higher levels primarily produced larval mortality (Su et al. 2003b, 2014). The
World Health Organization recommended novaluron for mosquito larval control
(WHO 2006) and also approved it for use
in drinking water.
Mosquiron CRDs have been evaluated
with success in multiple water bodies, but
only one earlier study has explored the
laboratory activity and field efficacy of this
product in storm drains or catch basins in
North America (Su et al. 2014). The product
label does not allow treatment of water directly connected to natural water bodies but
does permit the treatment of catch basins or
storm drains. Residual activity of novaluron
depends on water characteristics, such as
volume and presence of organic debris. Mosquiron CRDs are designed to deliver waxy
particles containing novaluron in a short period of time, while the active ingredient is released from the particle and adheres to the
surfaces of the storm drain and sediments
for continual release.
Twelve storm drains in downtown St. Augustine were selected for this study that contained approximately six inches (15.2 cm) of
standing water containing mosquito larvae
previously determined by dipper samples.
All drains were similar in size and structure.
Eight drains were used in the study. Four
drains were utilized for novaluron treatment. The remaining drains were untreated
controls located at a minimum of 30 meters
from the first treatment site. No untreated
control sites were connected to the treat-
ment storm water drainage lines, inflow, or
outflow, according to the storm water system
map provided by the City of St. Augustine.
No Bti treatments were made in the study
sites by AMCD or adjacent storm drains a
week prior to, or during, the study period.
One CRD was placed in each of the treatment storm drains according to the label.
The CRDs were cylindrical in shape, 5-10 cm
length by 1-2 cm diameter, and weighed 13.8
g/each. A 200 to 350 ml water sample was removed weekly from each drain using a white
plastic handheld-dipper (12.7 cm diameter
5.1 cm deep, Clarke ABC Dipper with Telescoping Handle, St. Charles, IL). Samples
from each drain were individually placed into
clear plastic mosquito emergence containers
(Bioquip, Rancho Dominguez, CA), labeled
by site location and date, then transported to
the AMCD insectary. If non-target organisms
were present in samples they were placed in
separate containers, with their sample water,
and any adverse effects recorded during the
experiment.
Mortality in aquatic stages was recorded
every 48 hours and adult mosquito inhibition weekly through 14 days. If no larvae
were present in storm drain samples then
ten laboratory-reared Cx. quinquefasciatus
early third instar larvae were added and recorded as above. Larvae in each emergence
container were maintained in AMCD’s insectary (12L:12D, 26.6°C, >70% RH) and
weekly fed approximately 0.03 g of ground
dog biscuits.
At post treatment week 4, retreatment
was necessary due to heavy rains that resulted in a considerable decrease in novaluron efficacy. In this trial 2, each of the eight
treatment drains (control included) were
treated. Four additional untreated drains
were used as controls. During trials 1 and 2
weekly precipitation, water level, and vegetation/debris contents were recorded for each
drain. The City of St. Augustine staff also collected salinity, dissolved oxygen, and pH in
the drains for the study from April to September 2014.
Statistical analysis ANOVA and a subsequent Student’s t test were run with JMP®
statistical software (SAS Institute Inc. 2013).
56
Average water temperature in the storm
drains was 23.2° C; salinity averaged 12.4
parts per thousand; dissolved oxygen averaged 0.41 mg/l; and the average pH was 6.8.
Salinity was the only measure that varied.
One week after treatment, novaluron produced an average of 73% adult inhibition
of Cx. quinquefasciatus (Figure 1). Efficacy
was affected by rainfall after week 1, which
appeared to have washed the CRD components out of the drain before they had time
to adhere to the sides of the drains. Adult
emergence was reduced to 23 and 33% for
weeks 2 and 3, respectively, but rose again to
78% at week 4. At that time, efficacy was unacceptable so drains were retreated on week
4. Retreatment on week 4 provided 98%
emergence inhibition in week 5 samples and
remained between 68 and 84% for the duration of the study.
Overall, there was a statistically significant difference in treatment versus control
in adult emergence inhibition (P = 0.002).
There was no significant difference between
treatments and controls in trial one (P =
0.05), while a statistically significant difference was found in trial two between treatment and controls (P = 0.004). Retreatment
on week 4 appeared to have boosted the
emergence inhibition despite the later rain
events (Figure 1). When larvae were found
in higher numbers during sampling, emergence inhibition was significantly greater
(P = 0.03). Similarly, when we focused on
trial two, lower numbers of larvae in the
drain samples exhibited higher emergence
inhibition and was statistically significant
(P = 0.02).
The City provided a storm drain cleaning schedule that allowed us to track which
drains had been cleaned during our study.
For drains cleaned most recently, the mean
emergence inhibition was higher than those
not cleaned recently or upon initiation of
the study (P = 0.04). Lower levels of vegetation and debris in drains resulted in statistically significant higher levels of emergence
inhibition (P = 0.04). Wax from the novaluron treatment was observed in multiple
treated drains post treatment on the sides
of the structures and on debris (especially
mulch chunks). When debris was minimal,
or non-existent, the wax appeared to adhere
more to the sides of the drains thereby increasing residual efficacy of the formulation.
The challenges to the study were the significant rain events.
We did not observe differences in nontarget mortality between treatments and
controls. Similarly, Su et al. (2003b) found
Figure 1. Weekly mean percent adult emergence inhibition of Culex quinquefasciatus after treatment of storm
drains by Mosquiron 0.12CRD (novaluron) at wks 0 and 4 and accompanying precipitation levels during the study.
Fulcher et al.: Field Evaluation of Mosquiron 0.12Crd Against Culex quinquefasciatus57
the novaluron 10% EC formula safe for nontargets in outdoor mesocosms at the application rate used for mosquito control. The
majority of non-targets in our study were
copepods and our observations were also
confirmed by Su et al. (2003b) and Arredondo-Jiminez and Valdez-Delgado (2006) that
these crustaceans are not adversely affected
by novaluron. Although we did not collect
larvivorus fish from the storm drains, Paiva
et al (2014) found 2% novaluron did not affect these predators. In addition, Tawatsin et
al. (2007) did not observe negative impacts
on fish when treating for Cx. quinquefasciatus
with 10% EC novaluron in polluted water.
Our average emergence inhibition of Cx.
quinquefasciauts with the CRD formulation
of novaluron was not as high as reported
by Arredondo-Jiminez and Valdez-Delgado
(2006) and Su et al. (2003b) who used a
10% EC formulation. Other studies, such as
Jambulingam et al. (2009), involving different formulations of novaluron found high
emergence inhibition for Cx. quinquefasciatus in wells. Additionally, Mulla et al (2003)
found that 10% EC and technical material
had long term activity in controlling Ae. aegypti in storage containers. However, our
second treatment with the CRD formulation
produced greater inhibition and may have
benefited from the previous application.
In summary, Mosquiron CRDs can be an
effective control agent against the production of adult mosquitoes in recently cleaned
storm drains over the course of multiple
weeks. As the City of St. Augustine continues to follow Best Management Practices for
urban storm water runoff, working in conjunction with AMCD in reducing the abundance of Cx. quinquefasciatus in these systems
will benefit the general public, at large, by
reducing the nuisance and risk of disease
transmission from this species.
This is a research report only, AMCD
does not endorse any commercial products
mentioned. We would like to thank John Regan, Todd Grant, Reuben Franklin, Glabra
Skipp, and Jeanne Moeller for their invalu-
able support and collaboration. Also, we
thank Barry Tyler (Pest Alto Environmental
Products Inc., Guelph, ON, Canada) for providing partial funding and product samples.
The assistance of Tom Downey, Ken Daniels,
Emily Thomson, and Rick Stockley in the
field during this evaluation was appreciated.
V. REFERENCES CITED
Arredondo-Jimenez, J. I. and K. M. Valdez-Degado.
2006. Effect of novaluron (Rimon 10EC) on the
mosquitoes Anopheles albimanus, Anopheles pseudopunctipennis, Aedes aegypti, Aedes albopictus, and Culex
quinquefasciatus from Chiapas, Mexico. Med. Vet.
Entomol. 20:377-387.
Jambulingam, P., C. Sadanandane, N. Nithiyananthan,
S. Subramanian, and M. Zaim. 2009. Efficacy of novaluron against Culex quinquefasciatus in small- and
medium-scale trials, India. J. Am. Mosq. Control Assoc. 25:315-322.
Metzger, M.E. 2004. Managing Mosquitoes in Storm water
Treatment Devices. University of California, Division of
Agriculture and Natural Resources. Publication 8125.
Miladin, S. 2004. Mosquito management going underground. Wing Beats. 15:10-12.
Moulis, R. A., H. B. Lewandowski, Jr, J. D. Russell, J. L.
Heusel , L.F.A.W. Peaty, D.G. Mead and R. Kelly.
2013. West Nile Virus activity in Chatham County,
Georgia in 2011. Wing Beats. 24: 23-27.
Mulla, M. S., U. Thavara, A. Tawatsin, J. Chompoosri,
M., Zaim, and T. Su. 2003. Laboratory and field evaluation of novaluron, a new acylurea insect growth
regulator, against Aedes aegypti (Diptera: Culicidae).
J. Vector Ecol. 28:241-254.
Müller, G. C., A. Junnila, W. A. Qualls, E. E. Revay, D. L.
Kline, S. Allan, Y. Schlein and R.D. Xue. 2010. Control of Culex quinquefasciatus in a storm drain system
in Florida using attractive toxic sugar baits. Med.
Vet. Entomol. 24:346-351.
Paiva, C. N., J. W. Lima, S. S. Camelo, C.de.F Lima, and
L. P. Cavalcanti. 2014. Survival of larvivorous fish
used for biological control of Aedes aegypti (Diptera:
Culicidae) combined with different larvicides. Trop.
Med. Intl. Health. 19:1082-6.
Rey, J. R., G. F. O’Meara, S. M. O’Connell, and M. M.
Cutwa-Francis. 2006. Factors affecting mosquito production from storm water drains and catch basins in
two Florida cities. J. Vector Ecol. 45:384-390.
SAS Institute, Inc. 2013. JMP, Version 11.1 Cary, NC.
Strickman, D and J. T. Lang. 1986. Activity of Culex quinquefasciatus in an underground storm drain in San Antonio, Texas. J. Am. Mosq. Control Assoc. 2:379-381.
Su, T., J. P. Webb, R. P. Myer and M. S. Mulla. 2003a.
Spatial and temporal distribution of mosquitoes in
underground storm drain systems in Orange County, California. J. Vector Ecol. 28: 79-89.
Su, T, M. S. Mulla, and M. Zaim. 2003b. Laboratory and
field evaluations of novaluron, a new insect growth
regulator (IGR), against Culex mosquitoes. J. Am.
Su, T., M. L. Cheng, A. Melgoza, and J. Thieme. 2014. Laboratory and field evaluations of mosquiron 0.12CRD, a
new formulation of novaluron, against Culex mosquitoes. J. Am. Mosq. Control Assoc. 30:284-290.
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Tawatsin, A., U. Thavara, P. Bhakdeenuan, J. Chompoosri, P. Siriyasatien, P. Asavadachanukorn, and
M. S. Mulla. 2007. Field evaluation of novaluron, a
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POPULATION REDUCTION OF MOSQUITOES AND BITING
MIDGES AFTER DEPLOYMENT OF MOSQUITO MAGNET®
TRAPS AT A GOLF COURSE ADJACENT TO SALTMARSH
HABITATS IN ST. AUGUSTINE BEACH, FLORIDA
RUI-DE XUE, WHITNEY A. QUALLS¹ AND DANIEL L. KLINE2
500 Old Beach Rd, St. Augustine, FL 32080
Department of Public Health Sciences
Miller School of Medicine
University of Miami, Miami, FL 33124
2
Center for Medical, Agricultural, and Veterinary Entomology
Gainesville, FL 32608
2
ABSTRACT. A two-year study was conducted
to evaluate the effectiveness of Mosquito Magnet® Liberty Plus (MMLP) traps to reduce
mosquito and biting midge abundance in a
St. Augustine Beach, FL golf community bordered by salt marsh habitat. These traps were
placed in residential backyards on the fringe
of the salt marsh for the purpose of reducing
the number of mosquitoes and biting midges
entering the central residential area. These
traps collected a total of 35,265 mosquitoes
from 9 genera where Aedes taeniorhynchus
(28%), and Anopheles crucians (27%) were
the two major species. Mosquito Magnet X
(MMX) traps were used to monitor mosquito
and biting midge population abundance in
the residential backyards. These traps collected 8,998 mosquitoes from 5 genera with Culex
nigripalpus (75%) as the major species. More
than one million biting midges were collected from both residential areas with Culicoides
furens (95%) being the dominant species.
Mosquito and biting midge abundance from
the MMX traps in residential backyards were
significantly reduced by 59 and 23%, respectively, compared with trap abundance from
the residential areas with MMLP traps in the
adjacent salt marsh. However, mosquito landing rates were not significantly reduced in the
residential areas.
Mosquito Magnet Liberty Plus trap, Mosquito Magnet X traps, biting midges
I. INTRODUCTION
Mosquito traps combined with attractants
have been evaluated by several researchers
and remain the standard for adult mosquito
surveillance (Allen et al. 2009; Chaves et al.
2014; Jackson et al. 2012). Recently, many
commercial mosquito traps have integrated
attractants into their units for the express
purpose of providing control of adult mosquitoes (Kline 2006, Xue et al. 2008). These
types of traps have attracted homeowner
attention due to safety concerns regarding
pesticide-based mosquito control technology. A variety of traps and lures that emit
host cues have been marketed for the general public. A prime example is the Mosquito
Magnet® mosquito trap series (Woodstream
Corp., Lititz, PA) that has been marketed
worldwide to consumers as a convenient, effective method to control adult mosquitoes
outdoors.
Mosquito Magnet traps have been evaluated against a variety of nuisance mosquitoes
(Henderson et al. 2006; Jackson et al. 2012;
Kitau et al. 2010; Smith et al. 2010) and biting midges (Cilek and Hallmon 2005; Lloyd
et al. 2008). However, these traps have not
been evaluated for their effectiveness at re-
Key Words. Aedes taeniorhynchus, Anopheles
crucians, Culex nigripalpus, Culicoides furens,
59
60
ducing major mosquito and biting midge
populations in large residential/recreational areas (such as golf club communities)
with surrounding salt marsh habitat.
The purposes of this study were to: (1)
determine species composition of mosquitoes and biting midges in the salt marsh
habitat and central residential area in a large
golf course community, (2) evaluate whether Mosquito Magnet Liberty Plus traps can
reduce population abundance of the major
mosquito and biting midge species encountered in the residential areas, compared with
the salt marsh habitat, and (3) determine
whether traps can reduce human landing
rate counts of mosquitoes in the residential
area.
This study was conducted from July to
October, 2004-2005, at the Marsh Creek
Golf Course on Anastasia Island, St. Augustine Beach, Florida. This golf course covers
9,333 acres that includes a residential community with about 400 homes. The habitat
surrounding the golf course is primarily
salt marsh with low-lying grassy areas that
will retain rainwater for days at a time. The
central residential area and residential backyards adjacent to the salt marsh were each
divided into 2 sections. In one area, 16 Mosquito Magnet Liberty Plus (MMLP) traps
(American Biophysics Corporation), baited
with octenol, were placed at 500 m intervals
in central residential backyards. The same
number of traps, and set up, were placed in
the backyard of homes adjacent to the salt
marsh. Residences with MMLP traps will be
referred to as “treatments” and were operated from Monday through Thursday for
4 nights per week. Controls consisted of
homes in each residential area without traps.
Treatment and control areas were switched
back and forth weekly, within each residential area, during the 6 wk study. In order to
obtain additional information of mosquito
abundance in the central residential section
with MMLP traps, landing rate counts (LRC)
were conducted (2 m away from each trap)
by a volunteer who recorded the number of
mosquitoes landing on both forearms for 3
minutes at the time of MMLP collection.
In order to monitor temporal mosquito
and biting midge abundance in the treatment and control sites, a MMX trap was
placed in each of two backyards in all areas.
Traps were suspended by a shepherd’s hook
and baited with dry ice (Xue et al. 2008).
Distance between MMX traps in each area
was about 1.5 km while the distance between
MMX traps and MMLP traps ranged from
1-3 km. MMX traps were turned on a Thursday and collected 24 h later on Friday. Mosquito and biting midge collections from all
MMLP and MMX traps were transported to
the laboratory for species identification and
counting under a 10-40X-zoom dissection
microscope.
Statistical analysis. All statistical analyses
were performed using SAS (SAS Institute
2001). Prior to analyses, mean trap counts
were log (x+1) transformed then subjected
to PROC MIXED with a repeated measures
analysis of variance (ANOVA). A χ2 test was
performed on mosquito collections to determine if differences existed between treatment and control areas (P<0.05).
III. RESULTS
A total of 35,265 mosquitoes representing 27 species from 9 genera were collected
from all MMLP traps from backyard residences adjacent to the salt marsh. The species composition is presented in Table 1. The
major species of mosquitoes in this area was
Aedes taeniorhynchus (Wiedemann) (28%),
Anopheles crucians Wiedemann (27%), Culex
nigripalpus Theobald (12%), Psorophora columbiae (Dyar and Knab) (9%), Aedes sollicitans (Walker) (6%), and Ae. infirmatus (Dyar
and Knab) (5%). Total abundance from all
MMLP traps in the central residential section totaled 8,998 mosquitoes and contained
10 species from 6 genera. The dominant species in these traps was Cx. nigripalpus (75%)
(Table 2). Collections of all MMX traps from
residential areas adjacent to the salt marsh
(with MMLP traps) totaled 27,072 mosquitoes and 495,780 biting midges (95% Culicoides furens and 5% C. mississippiensis) while
Xue et al.: Population Reduction of Mosquitoes and Biting Midges
Table 1. Species composition of saltmarsh and freshwater mosquitoes collected by Mosquito Magnet Liberty
Plus traps baited with octenol in residential backyards
adjacent to salt marshes and collected by Mosquito
Magnet X traps baited with dry ice in central residential areas, Marsh Creek golf course, St. Augustine, FL
2004-2006a.
Species
Aedes albopictus
Ae. atlanticus
Ae. canadensis
Ae. fulvus pallens
Ae. infirmatus
Ae. mathesoni
Ae. mitchellae
Ae. sollicitans
Ae. taeniorhynchus
Ae. triseriatus
Ae. vexans
Anopheles atropos
An. crucians
An. punctipennis
An. quadrimaculatus
Culex erraticus
Cx. nigripalpus
Cx. quinquefasciatus
Culiseta melanura
Coquillettidia perturbans
Mansonia dyari
Ms. titillans
Psorophora ciliata
Ps. columbiae
Ps. cyanescens
Ps. ferox
Ps. howardii
Central
Salt marsh side residential area
(%)
(%)
0.03
2
0.001
0.09
5
0.001
0.09
6
28
0.001
0.02
2
27
1.3
2.6
0.2
12
3
1.2
0.1
0.08
0.02
0.001
9
0.002
0.5
0.08
0.03
0.1
0
0
3
0
0
0
4
0
0
0
6
0
0.7
0
75
3
3
0
0
0
0
5
0
0
0
χ² = 4260.81, df = 26, P<0.01
a
45,579 adult mosquitoes and 2,195,533 biting midges were collected from MMX traps
in the same area without MMLP traps.
Mosquito and biting midge collections
from MMX traps in MMLP residential backyards adjacent to the salt marsh were significantly reduced by 59 and 23%, respectively,
compared with trap abundance in similar
areas without MMLPs. This trend continued
in the central residential area where MMLP
traps provided about a 63% reduction in
the number of mosquitoes in this area
compared with the same area without traps
(Table 2). Unfortunately, mosquito LRCs in
61
central residential and salt marsh areas with
MMLP traps were not significantly different
from LRCs in areas without traps (Table 3).
IV. DISCUSSION
Our study found that Mosquito Magnet
Liberty Plus traps significantly reduced mosquito and biting midge populations in the
central residential area of the Marsh Creek
Golf Course. Similar population reduction
has been observed by other in trap out studies where Mosquito Magnets have been evaluated (Cilek and Hallmon 2005, Kline 2002,
2007, Collier et al. 2006, Qualls and Mullen
2007, Lloyd et al. 2008, Li et al. 2010). Unfortunately, Mosquito Magnet Liberty traps
placed in residential backyards adjacent to
the salt marsh or within the central residential of the golf course did not result in a significant reduction of landing rate counts,
compared with areas without MMLPs. It is
possible that the sites where landing rates
were conducted were too close (2 meters)
to the MMLP sites where more mosquitoes
may have been present. However, MMLPs
did collect a large number of mosquitoes
that had significantly reduced the number
of mosquitoes in those areas.
The deployment of a linear array of Mosquito Magnet traps in residential backyards
that are adjacent to salt marsh areas may be
considered to be an environmentally friendly method to reduce the nuisance mosquito
and biting midge problems, especially in sensitive habitats. This control method could be
easily adopted and accepted by the residents
who live by salt marshes.
V. ACKNOWLEDGEMENTS
The authors wish to thank Alex Santoro
and James Wynn for their assistance on the
project. We also thank A. Grant and K. McKenzie from the former American Biophysics Corporation for providing the Mosquito
Magnet Liberty Plus and Mosquito MagnetX traps. We also the May Management Company and many residents in Marsh Creek to
allow us to use their properties during the
study. This is a research report only and
62
Table 2. Total numbers of mosquitoes collected by MMX traps baited with dry ice in the central residential areas
between treated areas with Mosquito Magnet Liberty Plus (MMLP) traps and untreated control areas (without
MMLP traps), Marsh Creek, St. Augustine, Florida 2004-2006a.
Species
Central area in treated section
Aedes albopictus
Ae. atlanticus
Ae. infirmatus
Ae. taeniorhynchus
Anopheles crucians
An. quadrimaculatus
Culex nigripalpus
Cx. quinquefasciatus
Culiseta melanura
Psorophora columbiae
Total
1
4
166
138
226
18
2,703
100
14
105
3,475
Central area in untreated control section
2
5
122
199
310
41
4,049
190
259
346
5,523
χ² = 231.081, df = 9, P < 0.01
a
Table 3. Mean human landing rate counts of Aedes taeniorhynchus (primarily) between Mosquito Magnet Liberty
Plus traps-treated section and untreated (control) section, Marsh Creek, St. Augustine, Florida 2004-2005a.
Years
Treated
salt marsh area
2004
2005
Total
0.52
2.02
2.54
Untreated control
Central residential area
Central residential area (MMX site)
salt marsh area (MMX site) in treated section
in untreated control section
1.36
1.83
3.19
0.88
2.13
3.02
1.00
2.25
3.25
χ² = 1.005, df = 3, P>0.05
a
mention of specific products does not indicate endorsement by the AMCD. Volunteers
gave informed consent prior to participation in the study and this research study was
conducted according to protocol number
AMCD 10-13-2005 as approved by the Anastasia Mosquito Control Board of Commissioners for use of human subjects in operational projects.
Allen, R.A., C.N. Lewis, M.V. Meisch, and D.A. Dame.
2009. Comparison of three residential mosquito
traps against riceland mosquitoes in southeast Arkansas. J. Am. Mosq. Control Assoc. 25:110-112.
Chaves, L.S., G.Z. Laporta, and M.A. Sallum. 2014. Effectiveness of Mosquito Magnet in preserved area on
the coastal Atlantic rainforest: implication for entomological surveillance. J. Med. Entomol. 51:915-924.
Cilek, J.E .and C.F. Hallmon. 2005. The effectiveness
of the Mosquito Magnet trap for reducing biting
midge (Diptera: Ceratopogonidae) populations in
coastal residential backyards, J. Am. Mosq. Control
Assoc. 21:218-221.
Collier, B.W., M.J. Perich, G.J. Boquin, S.R. Harrington,
and M.J. Francis. 2006. Field evaluation of mosquito
control devices in southern Louisiana. J. Am. Mosq.
Henderson, J.P., R. Westwood, and T. Galloway. 2006.
An assessment of the effectiveness of the Mosquito
Magnet Pro model for suppression of nuisance mosquitoes. J. Am. Mosq. Control Assoc. 22:401-407.
Jackson, M.J., J.L. Gow, M.J. Evelyn, T.J. McMahon, T.J.
Howay, H. Campbell, J. Blancard, and A. Thielman.
2012. An evaluation of the effectiveness of a commercial mechanical trap to reduce abundance of
adult nuisance mosquito populations. J. Am. Mosq.
Kitau, J, H. Pates, T.R. Rwegoshora, D. Rwegoshore, J.
Matowo, E.J., Kweka,, F.W. Mosha, K. McKenzier,
and S.M. Magesa. 2010. The effect of Mosquito Magnet Liberty Plus trap on the human mosquito biting
rate under semi-field conditions. J. Am. Mosq. Control Assoc. 26:287-294.
Kline, D.L. 1999. Comparison of two American Biophysics mosquito traps: the professional and a new
counterflow geometry design. J. Am. Mosq. Control
Assoc. 15:276-282.
Kline, D.L. 2002. Evaluation of various models of propane powered mosquito traps. J. Vect. Ecol. 27:1-7.
Kline, D.L. 2006. Traps and trapping techniques for
22:490-496.
Kline, D.L. 2007. Semiochemicals, traps/targets and
mass trapping technology for mosquito management. J. Am. Mosq. Control Assoc. 23(2S):241-251.
Li, C.X., Y.D. Dong, X.L. Zhang, C. Chen, S.P. Song, B.
Deng, T.Y. Zhao, and R.D. Xue, 2010. Evaluation of
octenol and lurex as baits in Mosquito Magnet Pro
traps to collect vector mosquitoes in China. J. Am.
Xue et al.: Population Reduction of Mosquitoes and Biting Midges
Lloyd, A.M., D.L. Kline, J.A. Hogsette, P.E. Kaufman, and
S.A. Allan. 2008. Evaluation of two commercial traps
for the collection of Culicoides (Diptera: Ceratopogonidae). J. Am. Mosq. Control Assoc. 24:253-262.
Qualls, W.A. and G.R. Mullen. 2007. Evaluation of the
Mosquito Magnet Pro trap with and without 1-octen-
63
3-ol for collecting Aedes albopictus and other urban
mosquitoes. J. Am. Mosq. Control Assoc. 23:131-136.
Xue, R.D., M.A. Doyle, and D.L. Kline. 2008. Field
evaluation of CDC and Mosquito Magnet® X traps
baited with dry ice, sachet, and octenol against mosquitoes. J. Am. Mosq. Control Assoc. 24:249-252.
SUBLETHAL DOSES OF AN ATTRACTIVE TOXIC SUGAR
BAIT MIXED WITH THE INSECT GROWTH REGULATOR
PYRIPROXYFEN DID NOT EFFECT SURVIVAL OR
FECUNDITY OF AEDES AEGYPTI
CODI S. ANDERSON, JODI M. SCOTT, ALICE P. FULCHER,
GUNTER C. MULLER¹, AND RUI-DE XUE
Department of Microbiology and Molecular Genetics, IMRIC
Kuvin Centre for the Study of Infectious and Tropical Diseases
Faculty of Medicine, Hebrew University, Jerusalem, Israel 91120
1
ABSTRACT. Addition of the insect growth
regulator, pyriproxyfen (0.5%), into an attractive toxic sugar bait solution containing
a sublethal dose of eugenol (0.0125%) or
boric acid (0.1%) had no significant effect
on the survival or fecundity of adult Aedes
aegypti compared with sugar baits without
these active ingredients.
laayoune et al. 2013). In addition, boric acid
and eugenol have been reported to be orally
effective against several species of mosquitoes (Xue and Barnard 2003, Qualls et al.
2014, Xue et al. 2006). Interestingly, Ali et al.
(2006) reported that sub-lethal doses of boric acid affected survival, blood feeding, and
fecundity in Aedes albopictus (Skuse) when
orally ingested. A recent laboratory study by
Fulcher et al. (2014) reported effective control of adult and larval mosquitoes in runoff
water from plants previously sprayed with
an attractive sugar bait (ASB) and the insect
growth regulator (IGR) pyriproxyfen. In addition, Chism and Apperson (2003) demonstrated 38% emergence inhibition when
pyriproxyfen-treated Ae. albopictus oviposited
in untreated larval habitats. Therefore, we
decided to investigate the effects of sub-lethal ATSBs containing boric acid or eugenol
in combination with pyriproxyfen on survival, fecundity, and fertility of the dengue and
chikungunya vector, Aedes aegypti (L.).
Key Words. Aedes aegypti, pyriproxyfen, eugenol, boric acid, attractive toxic sugar bait
I. INTRODUCTION
Female and male mosquitoes feed on
plant nectars as carbohydrate sources in
order to obtain the necessary energy requirements to survive. Indeed, Marshall et
al. (2013) found, in laboratory studies, that
some female Anopheles species will feed on
sucrose daily. A number of authors have reported on the evaluation of a variety of plant
sugar sources that could be incorporated
into toxic baits thereby exploiting a mosquito’s sugar feeding behavior (Muller et al.
2008, Muller et al. 2010, Beier et al. 2012)
Studies of attractive toxic sugar baits
(ATSB), during the past decade, have focused predominantly on integrating topically active insecticides such as pyrethroids,
neonicotinoids, or organophosphates into
oral formulations for adult mosquito control (Allan 2011, Muller et al. 2010, Khal-
Mosquitoes. The Ae. aegypti colony used in
our study, was obtained from the USDA, Center for Medical, Agricultural, and Veterinary
Entomology, Gainesville, FL. Oviposition paper containing eggs of this species was cut
into 2 cm strips and placed into larval rearing
pans (28 cm × 56 cm) containing 2,500 ml of
reverse osmosis water (RO). This method was
64
Anderson et al.: Sublethal Doses of An Attractive Toxic Sugar Bait
utilized to produce a maximum of 500 mosquito larvae per pan. Mosquito larvae were
given 3 mg of 3:2 ratio of bovine liver powder and brewer’s yeast (ICN Biomedical, Inc.,
OH). While in the pupal stage, mosquitoes
were placed into 15 cm × 15 cm × 5 cm Ziploc
brand sandwich containers (SC Johnson &
Son Inc., Racine, WI) and moved into rearing cages (Bioquip Products Inc., Compton,
CA) to emerge. Cotton balls saturated with
10% sucrose solution in 266 ml clear plastic
cups (SOLO®, Dart Container Corporation,
Mason, MI) were provided ad libitum. Adult
mosquitoes were utilized in bioassays once
they reached 5-7 days of age.
Attractive sugar bait (ASB). Mango fruit
pulp was combined with white cane sugar
at a 1:1 ratio with approximately 0.8% lime
juice. The mixture was stirred and heated to
100° C and allowed to cool to 26° C, producing mango syrup. To prepare the attractive
sugar bait (ASB), the syrup was mixed with
reverse osmosis water at a ratio of 1:3. Green
food coloring (McCormick brand) was added (0.5%) to the formulation in order to visualize ingestion by the mosquitoes (Müller
et al. 2010). Also about 0.08% Agricultural
Poly Control 2 Sticker Spreader (Brewer
International, Vero Beach FL) was added
to the ASB solution because this substance
was previously used during previous field applications of ATSB (JM Scott unpubl. data).
Attractive toxic sugar bait was prepared by
adding toxicant to the ASB (percentages determined below).
ATSB sub-lethal determination. As reported
by Ali et al. (2006), we selected 0.1% boric
acid as the sub-lethal concentration for testing this chemical. Because biologically active
sub-lethal concentrations of eugenol have
not been reported, we determined this concentration is a series of ASB serial dilution
bioassays. The following procedures were
followed: nine to twelve female Ae. aegypti
(5-7 days old) were mouth aspirated into individual 1 L glass mason jars, with the mouth
of the jars covered with Wal-Mart brand tulle
mesh secured with the metal bands of the
jar lid. A total of 25 jars were used for each
of 3 repetitions. Negative controls were fed
ASB only. Eugenol concentrations in ASB
65
solution were 0.1%, 0.05%, 0.025%, and
0.0125%. Green food coloring was not added to these solutions. All mosquitoes were
fed about 12 ml of each solution through
cotton balls placed on top of the tulle mesh.
Cotton balls remained on the mesh as adult
mortality was recorded at 24, 48, and 72
hours. The experiment was repeated three
times. Based on the results of the eugenol
bioassays, 0.0125% was selected as the sublethal concentration for incorporation into
ATSBs (Figure 1).
ATSB/IGR bioassays. Boric acid (0.1%)
and eugenol (0.0125%) ATSBs were separately mixed with 0.5% pyriproxyfen (P)
(NyGuard® [10% AI] IGR Concentrate,
Minneapolis, MN) and 0.5% P with ASB only,
as the final formulations for testing. Control formulations consisted of ASB without
a toxicant (negative control) and ATSB 1%
boric acid (positive control). Approximately
70-80, five to seven-day old male and female
mosquitoes were mouth aspirated into modified 5 L buckets with a 15 cm hole cut into
the side covered by a fabric sleeve adhered
with glue (Qualls et al. 2014). This sleeve
allowed for continuous access to the mosquitoes, with minimal escapees. The bucket
opening was covered with Wal-Mart brand
tulle mesh secured with rubber bands. Each
treatment was assigned randomly to each of
five buckets. Baits were poured into unsalted
natural collagen sausage casings, and then
tied with a balloon (single) knot. Bait “sausages” were placed on top of the tulle mesh,
and mosquitoes were allowed to feed for 48
hours. Adult mortality was recorded at 24
and 48 h.
Fecundity. Males and females (70-80 individuals, five to seven-day old) were placed
together in each bucket (to ensure mating
occurred) and presented with bovine blood
(36° C) for 48 hours administered in the same
fashion as the bait sausages. After this time,
blood-fed individuals were mouth aspirated
from the buckets into oviposition cages (that
consisted of inverted 266 mL clear plastic
cups with a 1 cm hole in the base to facilitate
transfer) at the rate of one mosquito per cup.
Ten cups were used per treatment including
controls. This hole was stoppered with cot-
66
(F = 8.757, df = 4,45, P < 0.001)
1
Figure 1. Average percent mortality of adult Aedes aegypti to various concentrations of eugenol when mixed with
an attractive sugar bait (ASB) solution after ingestion at 24, 48, and 72 h1. ASB without eugenol is an untreated
control.
ton and utilized throughout the experiment
to administer sucrose solution (10%) to the
mosquitoes. The mouth of the cup was covered with tulle and secured into place with a
rubber band. A 2 cm × 6 cm strip of germination paper was placed loosely into the mouth
of the inverted cup by stretching the rubber
band to the side and sliding the paper inside
to rest on the mesh.
The oviposition cages were placed at a
45° angle into larval rearing pans (28 cm ×
56 cm) that contained 1 cm of reverse osmosis water to allow the oviposition paper to be
dampened. Pans were placed into an incubator and maintained at 26.6° C and 14:10 L:D.
This study was repeated three times with 10
cups (females) per control/treatment and
50 cups total per repetition. Mosquitoes
were allowed to oviposit for 48 hours. After
this time, egg papers were collected, number of eggs counted, and allowed to dry for
five days allowing for egg embryonation.
Fertility. Eggs were hatched in 400 ml of
water in 15 cm × 15 cm × 5 cm plastic Ziploc® brand sandwich containers with lids
and will be referred to as “hatch containers”.
Larval food was added and lids left unsealed
on one corner in order to allow for ventilation. Hatch containers were observed daily
for pupae. Pupae were counted and transferred to clear plastic 266 ml plastic cups
through a 0.5 cm hole in the tulle mesh covering the aperture of the cup. There were
50 cups per repetition. When not in use,
the hole was covered with tape. Emerging
adults were provided with a cotton ball saturated with 10% sucrose solution, ad libitum,
placed on top of the mesh. Successful emergence was counted by subtracting the dead/
un-emerged pupae from total pupae. Adult
mortality was recorded at 48 h post initial
oviposition and observed for ten days.
Statistical analysis. Analysis of data used
JMP statistical software (SAS Institute, Inc.
2012). Mean adult survival was assessed
through chi square analysis to determine significant differences between formulations.
Blood feeding, and total egg data were subjected to ANOVA to determine significant
differences between formulations. For all
analyses, differences were considered significant when P <0.05.
Adult Survival. The ASB+P 1% boric acid
formulation caused significantly greater
mortality of Ae. aegypti at 48 h post ingestion,
compared with the mortality caused by other formulations (Figure 2). Overall, none of
the other ATSBs significantly affected mortality or survival of females, 10 days after oviposition, even though the 0.1% boric acid
formulation resulted in >16% mortality (Figure 3). However, Ali et al. (2006) reported
that a 0.1% boric acid sugar bait significantly
reduced the survival rate of Ae. albopictus
compared with an untreated control. We believe that this may be caused, in our study,
by lower boric acid concentration in the ASB
after mixture with pyriproxyfen.
In our initial bioassay identification of a
eugenol sub-lethal concentration suitable
67
for this study, we found that 0.1% produced
significantly greater mortality compared with
0.0125% selected for our sublethal evaluations (Figure 3). These results were similar
with those of Qualls et al. (2014). However,
Hao et al. (2008, 2012) found that eugenol at
higher concentrations significantly reduced
blood feeding in adult mosquitoes and might
act as a repellent. Perhaps, this may be the
reason, in our study, why eugenol mixed with
pyriproxyfen did not result in any significant
mortality compared with controls.
Fecundity and fertility. No significant difference was observed between the total number of eggs laid between ATSBs and controls
(Figure 4). This result was similar with the
report that sublethal concentrations of boric
acid sugar bait significantly reduced fecundity of Ae. albopictus (Ali et al. 2006), but the
reduction of number of eggs in our experiment was not significant, compared with the
other baits. As stated earlier, this may have
been caused by the addition of pyriproxyfen
24 h (χ² = 11.753, df = 4, P<0.01) and 48 h (χ² = 17.108, df = 4, P<0.01)
1
Figure 2. Average percent mortality of adult Aedes aegypti from ingestion of an attractive sugar bait (ASB) mixed
with either boric acid or eugenol with 0.5% pyriproxyfen (P) after 24 and 48 h. ASB without boric acid or eugenol
is an untreated control1.
68
(F = 0.269, df = 4,20, P > 0.05)
1
Figure 3. Average percent mortality of Aedes aegypti at 10 days following oviposition after feeding on attractive
sugar baits (ASB) containing either boric acid or eugenol with pyriproxyfen (P). ASB without boric acid or eugenol
is an untreated control1.
to the ATSB that reduced the concentration
of boric acid. However, Chism and Apperson
(2003) did not observe any adverse effects on
fecundity even when gravid Ae. albopictus were
exposed to pyriproxyfen-treated surfaces.
In our study, egg hatchability considerably varied between ATSBs. In one repetition of the experiment, complete larval
emergence of all eggs occurred. In another repetition, three of 25 containers with
eggs did not produce larvae. In other repetitions, only a single larva was recovered
from eggs produced from females that fed
upon ASB+P (0.5%), ASB+P+(0.0125%)
eugenol, and ASB+P+(0.1%) boric acid.
Larvae in another container, from another
repetition, died in the third instar stage
(ASB+P (0.5%). We believe that the containers were possibly contaminated by an
unknown contaminant, or contamination
was the result of sucrose solution cotton
balls where eggs had been oviposited during the evaluation.
In summary, sublethal concentrations of
boric acid or eugenol in an ASB mixed with
0.5% pyriproxyfen did not significantly affect survival of Ae. aegypti. Also, mixing pyriproxyfen with boric acid or eugenol as baits
at the concentrations used in this study did
not reveal vertical transfer nor produce any
significant impact on fecundity by this insect
growth regulator.
Thank you to the fellow interns and Anastasia Mosquito Control District employees
who volunteered in assisting with this project and to AMCD for the opportunity to conduct the study.
V. REFERENCES CITED
Ali, A. R. D. Xue, and D. R. Barnard. 2006. Effects of
sublethal exposure to boric acid sugar bait on adult
survival, host-seeking, blood feeding behavior, and
reproduction of Aedes (Stegomyia) albopictus. J. Amer.
Allan, S. A. 2011. Susceptibility of adult mosquitoes to
insecticides in aqueous sucrose baits. J. Vect. Ecol.
36:59-67.
Beier, J. C., G. C. Muller, G. U. Weidong, K. L. Arheart,
and Y. Schlein. 2012. Attractive toxic sugar bait
1
69
(F = 0.773, df = 4,20, P > 0.05)
Figure 4. Total number of eggs laid by Aedes aegypti after ingestion of attractive sugar baits (ASB) containing
either boric acid or eugenol with pyriproxyfen (P). ASB without boric acid or eugenol is an untreated control1.
(ATSB) decimates populations of Anopheles malaria
vectors in arid environments regardless of local
availability of favored sugar-source blossoms. Malar.
J. 11:31.
Cheng, S. S., J. Y. Liu, K. H. Tsai, W.J. Chen, and S. T.
Chang. 2004. Chemical composition and mosquito
larvicidal activity of essential oils from leaves of different Cinnamomum osmopheloeum provenances. J.
Agri. Food Chem. 52:4395-400.
Chism, B. D. and C. S. Apperson. 2003. Horizontal
transfer of the insect growth regulator pyriproxifen
to larval microcosms by gravid Aedes albopictus and
Ochleotatus triseriatus mosquitoes in the laboratory.
Fulcher, A., J. M. Scott, W. A. Qualls, G. C. Muller, and
R.D. Xue. 2014. Attractive toxic sugar baits mixed
with pyriproxyfen sprayed on plants against adult
and larval Aedes albopictus (Diptera: Culicidae). J.
and blood-feeding behavior of Aedes albopictus (Diptera: Culicidae) exposed to vapors of geraniol, citral,
citronellal, eugenol, or anisaldehyde. J. Med. Entomol. 45:533-539.
Hao, H., J. Sun, and J. Dai. 2012. Preliminary analysis
of several attractants and spatial repellants for the
mosquito, Aedes albopictus using an olfactometer. J.
Ins. Sci. 12:1-10.
Khallaayoune, K., W. A. Qualls, E. E. Revay, S. A. Allan, K. L. Arheart, V. D. Kravchenko, R. D. Xue, Y.
Schlein, J. C. Beier, and G. C. Muller. 2013. Attractive toxic sugar baits: control of mosquitoes with the
low-risk active ingredient dinotefuran and potential
impacts on nontarget organisms in Morocco. Environ. Entomol. 42:1040-1045.
Marshall, J.M., M.T. White, A.C. Ghani, Y. Schlein, G.C.
Muller, and J.C. Beier. 2013. Quantifying the mosquito’s sweet tooth: modeling the effectiveness of attractive toxic sugar baits (ATSB) for malaria vector
control. Malaria J. 12:291-304.
Muller, G .C., V. D. Kravchenko, and Y. Schlein. 2008.
Decline of Anopheles sergentii and Aedes caspius populations following presentation of attractive toxic
(spinosad) sugar baits stations in an oasis. J. Am.
Mosq. Contr. Assoc. 24:147-149.
Muller, G. C., A. Junnila , W. A. Qualls, E.E. Revay, D. L.
Kline, S. A. Allan, Y. Schlein, and R. D. Xue. 2010.
Control of Culex quinquefasciatus in a storm drain
system in Florida using attractive toxic sugar baits.
Qualls, W. A., G. C. Muller, E. E. Revay, S. A. Allan, K.
L. Arheart, J. C. Beier., M. L. Smith, J. M. Scott., V.
D. Kravchenko, A. Hausmann, Z .A. Yefremova, and
R. D. Xue. 2014. Evaluation of attractive toxic sugar
bait (ATSB) – barrier control of vector and nuisance
mosquitoes and its effect of non-target organisms
in sub-tropical environments in Florida. Acta Trop.
131:104-110.
SAS Institute, Inc. 2012. JMP®, Version 10.1. Cary, NC.
Xue, R. D. and D. R. Barnard. 2003. Boric acid sugar
bait kills adult mosquitoes (Diptera: Culicidae). J.
Econ. Entomol. 96: 1559-1562.
Xue, R. D., D. L. Kline, A. Ali, and D. R. Barnard. 2006.
Application of boric acid to plant foliage for adult
mosquito control. J. Amer. Mosq. Control Assoc. 22:
297-500.
EVALUATION OF POWER BREEZER AND MISTING
CITRONELLA AGAINST AEDES ALBOPICTUS
EMILY H. THOMSON¹, JODI M. SCOTT1, ALI FULCHER1,
MICHAEL L. SMITH1, PHIL KOEHLER2, AND RUI-DE XUE1
500 Old Beach Rd, St. Augustine, FL32080
1
2
University of Florida, Entomology and Nematology Department,
University of Florida, Gainesville, FL 32611
ABSTRACT. An industrial high-velocity
misting fan, with and without a 1% citronella mixture, was evaluated for repelling
host seeking Aedes albopictus. Human participants were utilized as targets and arranged
in a horizontal line 5, 8, 10, and 15 meters
from the fan while mosquitoes were released
from the opposite end of the testing arena.
Control evaluations were conducted without
wind or citronella. Overall, no significant
difference in participant preference by Ae.
albopictus was observed in treatments or controls at any of the distances except at 8 m.
At 15 m, significantly more mosquitoes were
collected from participants with the fan on
(and no mist) compared with the control
(fan off). When the fan was misting 1% citronella, more mosquitoes landed on participants as distance from the fan unit increased
but no statistical significant difference between the control and treatment groups
occurred. In summary, the wind velocity
generated by the misting fan itself, or the citronella mixture, did not significantly reduce
host seeking Ae. albopictus and may not be a
practical method of protection from biting
mosquitoes for personal or public outdoor
events based upon the results of our study.
viruses, are leading global public health
concerns (Kilpatrick et al. 2006, Mitchell
1991). With the advancement of widespread human transportation, these diseases and their vectors are no longer endemically confined. In addition, as human
populations grow, especially in developing
countries, the risk of mosquito-borne illness is becoming more threatening (Kilpatrick et al. 2006). Aedes albopictus (Skuse)
is an important vector of several arboviral
pathogens and one that is competent in
transmitting many diseases in high density
urban and suburban settings because of
widely available larval developmental areas in close proximity to human habitation
(Mitchell 1991).
A recent report claiming that an oscillating table fan was capable of deterring
mosquito bites at a backyard gathering by
disrupting a mosquito’s ability to locate
potential hosts has provided additional
interest in non-chemical and natural repellent options (Broad 2013). Fans have
commonly been used to deter pest insects, such as cockroaches and fruit flies in
homes and businesses (Kolbe 2003). The
American Mosquito Control Association
describes most mosquitoes as weak fliers,
only capable of flying at 0.45 to 0.67 meters per second, and recommends placing
a large fan on a household deck or patio
to provide a “low tech solution” to reduce
mosquito bites (AMCA, 2014). Manipulating wind as a means of repellency is gaining more public attention and remains
relatively unstudied.
Key Words. Misting fan, Aedes albopictus,
citronella, host seeking
I. INTRODUCTION
Mosquito-borne illnesses, such as West
Nile, chikungunya, dengue, and yellow
70
Thomson et al.: Evaluation of Power Breezer and Misting Citronella Against Aedes albopictus71
Citronella oil is a widely recognized
and commonly utilized insect repellent.
Recent studies have debated the efficacy
of citronella to repel mosquitoes (Tawatsin et al. 2001, Yang and Ma 2005, Hao et
al. 2008, Gross and Coats 2015). Multiple
studies have hailed the success of combining wind currents with vapor or mists of
the repellent, DEET (Hoffman and Miller
2002, Hoffman and Miller 2003), the spatial repellent, metofluthrin in a clip-on fan
device (Xue et al 2012, Revay et al 2013),
or geraniol and citric acid in ALLCLEAR
Backyard Mosquito Misters (Revay et al
2013a) for use in reducing mosquito bites.
However, there is lack of evidence that commercial mechanical fans misting citronella
can repel the container-inhabiting mosquito, Ae. albopictus. Our study objectives
were to 1) determine the effectiveness of
a high powered commercial misting fan to
deter Ae. albopictus from host-seeking with
wind currents at multiple distances from
the machine, and 2) test the effectiveness
of the fan to prevent mosquito bites when
misting a 1% citronella oil at multiple distances from the machine.
Power Breezer® machines (Breezer
Holdings LLC, Deerfield Beach, Florida)
were acquired from the manufacturer
for testing. These industrial fans were
equipped with a 380 L (100 gal) tank and
locking rear casters to secure the fan in
place. Another feature that was utilized in
this study was the atomizer capable of producing a fine mist from the solution in the
tank. The unit was evaluated at Anastasia
Mosquito Control District (AMCD) district headquarters in a 12.2 × 18.3 m metal
garage previously cleaned and sealed for
use as the experimental arena. The Power
Breezer was situated in the northwest corner of the garage and oriented with the
fan-facing the opposing southeastern corner. A 24 m line was measured from the
�Figure 1. Orientation and average wind velocity at multiple distances of the Power Breezer machine during
experimental trials.
72
fan to the opposite corner of the building
with lines marked with blue painters tape
at 5 m, 8 m, 10 m, and 15 m (Figure 1).
For each distance, six points were marked
0.6 m from one another that represented
where each participant would stand for
evaluation.
Average wind speed of the Power
Breezer was determined at its highest
setting using a handheld SKYWATCH®
Xplorer 3 anemometer (JDC Electronics SA, Yverdon-les-Bains, Switzerland).
Speeds were recorded at angles left, right,
and center of the machine at the 5 m, 10
m, and 15 m distances. At 8 m, only wind
speed at the center point was measured.
Average wind-speeds are listed in Figure
1. The garage bay door closest to the unit
remained open to alleviate potential pressure buildup in the building generated by
the high-powered fan. The misting unit
flow rate averaged 100 ml/min for each
ten-minute trial.
Two hundred, caged, adult 5-7 day
old laboratory reared female Ae. albopictus were brought to the testing area and
placed at the opposite corner, 24 m from
the fan unit. For each test, mosquitoes
were allowed to acclimate to the new environment for up to 30 minutes. Six participants (3 males, 3 females) were used
in the study and did not rotate during
evaluations. Testing began at the farthest
distance from the fan (15 m). Mosquitoes
were released and given ten minutes to actively host seek participants. Mosquitoes
that came to the participants, during that
time were killed and counted. At the end
of the evaluation period total mosquito
landings were recorded. New mosquitoes
were used for each distance tested. Each
objective was evaluated only one time.
Objective one, evaluated the fan on
without mist, while in the second objective
the fan applied a 1% citronella oil in water mist. Testing distances were reduced to
the 10 m and 15 m points in the citronella
trials due to insufficient landing numbers
in previous trials at the 5 m and 8 m distances. Control trials were conducted with
the fan off and without citronella. Objec-
tive one was conducted in April of 2014,
while objective two was conducted in June
2014. During each objective, new participants were used for controls and treatments based on availability.
Statistical analysis All data were analyzed by ANOVA using JMP® software (SAS
Institute Inc. 2013). Overall, host seeking abundance of Ae. albopictus between
participants was compared within pooled
controls and pooled treatment groups to
determine if differences occurred. The
average number of mosquitoes able to locate participants in control (no wind) and
treatment wind trials (with and without citronella) at each distance from the Power
Breezer were compared. Significant differences were determined at p <0.05.
III. RESULTS
No significant difference in the number of Ae. albopictus recovered between participants at each sample distance occurred
with the exception of 8 m where significantly more mosquitoes were collected (15
m: F=0.1757, p=0.9620; 10 m: F=1.2236,
p=0.4005; 8 m: F=4.6032, p=0.0451; 5 m:
F=1.9314, p=0.2223) (Figure 2). During the
first objective, there were no observed difference between the treatments and controls
at 5, 8, 10 meters from the Power Breezer
fan (5 m: F=2.2321, p=0.1660, 8 m: F=0.0440,
p=0.8381, 10 m; F=2.4806, p=0.1463). At 15
m significantly more mosquitoes were collected from the treatment than the control
(F=23.416, p=0.0007) (Figure 2).
When the fan was misting 1% citronella, statistically more mosquitoes landed on
participants as distance from the fan unit
increased (F=18.2826, p=0.0003) (Figure
3). There were no observed difference
between treatments and control at either
distance from the Power Breezer fan (10
m: F=2.5966, p=0.1382 and 15 m: F=0.2062,
p=0.6595).
IV. DISCUSSION
The significant differences that we
found between the participants at 8 m in
Figure 2. Mean (±SE) host seeking Aedes albopictus collected at multiple distances from a Power Breezer. Treatment consisted of fan on with no mist, control consisted of the fan off.
control and treatment groups and at 15
m for the wind without citronella trial
may be attributable to a variety of random
variables within the testing arena. Time
interval between tests for each distance
and increased temperature in the partially
enclosed space may have influenced the
study results. In addition, trial participants
were also allowed to actively bend, squat,
and physically exhale carbon dioxide in
order to attract mosquitoes. At 15 m, participants were only 9 m from the mosquito
release site where the weakest winds farthest from the fan occurred. Low wind
Figure 3. Mean (±SE) host seeking Aedes albopictus collected at multiple distances from a Power Breezer. Treatment consisted of fan on with 1% citronella mist, control consisted of the fan off.
74
velocity from the fan may have facilitated
host-seeking behavior by enhancing dispersal of a participant’s kariomones making the mosquitoes to be more attractive
to their host.
Hoffman and Miller (2003) analyzed
the relationship of wind velocity with mosquito host seeking and flight behavior. The
authors reported that wind likely had the
greatest effect on host odor dilution for host
seeking mosquitoes. This mechanism outweighed the relationship of higher wind velocity, that can create intolerable flying conditions for the mosquito. The high velocity
wind stream produced by the Power Breezer
exceeded the 0.45 m/s flying capability of
mosquitoes at every test distance. But we
found that wind alone did not significantly
deter host seeking mosquitoes as pointed
out by Snow (1977) and Hoffman and Miller
(2002). We found that the wind stream from
the Power Breezer appeared to produce a
boundary layer of calmer conditions lower
to the ground and may have facilitated favorable conditions for mosquito up wind flight
to participants after host location.
In an earlier study, Hoffman and Miller
(2002) focused on manipulating wind currents while combining a DEET mist and
found this treatment provided temporary
protection from mosquitoes. These results
are in contrast with our study with citronella.
The differences in both studies may be attributable to the chemicals used and possibly concentrations. Most laboratory studies that test
repellency of citronella use concentrations
ranging from 10-30%, and often include 5%
vanillin to enhance repellency (Tawatsin et
al. 2001, Yang and Ma 2005, Hao et al. 2008)
while we only applied 1% citronella. In summary, wind velocity generated by the Power
Breezer fan itself, or with the citronella mixture, did not significantly reduce host seeking
Ae. albopictus and may not be a practical method for preventing mosquito bites for personal
during public outdoor events.
V. ACKNOWLEDGEMENTS
This is a research report only and
does not imply AMCD endorsement of
any commercial products. Volunteers gave
informed consent prior to participation
in the study. This study was conducted according to protocol number AMCD-10-132005 as approved by the AMCD Board of
Commissioners for use of human subjects
in the operation of projects.
AMCA (American Mosquito Control Association).
2014. Frequently asked questions [Internet].
Mount Laurel, NJ http://www.mosquito.org/
faq#fast [Accessed April 15, 2015]
Broad, W. J. 2013. A low-tech mosquito deterrent. The
New York Times. [Internet]. http://www.nytimes.
com/2013/07/16/science/a-low-tech-mosquito-deterrent.html?_r = 0 [accessed February 25, 2015].
Gross, A. D. and J. R. Coats. 2015. Can green chemistry provide effective repellents? In: Debboun M.,
Frances S. P., Strickman D.A., eds. Insect Repellents
Handbook. Second Edition. Boca Raton, Florida,
United States: CRC Press. pp. 75-90.
and blood-feeding behavior of Aedes albopictus
(Diptera: Culicidae) exposed to vapors of geraniol, citral, citronellal, eugenol, or anisaldehyde.
J. Med. Entomol. 45: 533-539.
Hoffman, E. J. and J. R. Miller. 2002. Reduction of
mosquito (Diptera: Culicidae) attacks on a human
subject by combination of wind and vapor-phase
DEET repellent. J. Med. Entomol. 39: 935-938.
Hoffman, E. J. and J. R. Miller. 2003. Reassessment of
the role and utility of wind in suppression of mosquito (Diptera: Culicidae) host finding: stimulus
dilution supported over flight limitation. J. Med.
Entomol. 40:607-614.
Kolbe, W. A. 2003. No hot air. Pest Control Technology.
31(6) http://www.pctonline.com/article/-Annual-FlyControl-Issue--No-Hot-Air. [Accessed 20 December
2014]
Kilpatrick, M., L. D. Kramer, M. J. Jones, P. P. Marra,
and P. Daszak. 2006. West Nile virus epidemics in
North American are driven by shifts in mosquito
feeding behavior. PloS Biology. 4:e82
Mitchell, C. J. 1991. Vector competence of North and
South American strains of Aedes albopictus for certain arboviruses: a review. J. Am. Mosq. Control
Assoc. 7:446-452.
Revay, E. E., A. W. A. Qualls, R. D. Xue, and G. C.
Muller 2013a. Evaluation of a portable backyard
mosquito mister against Culex pipiens and Aedes
albopictus in Israel. Tech. Bull. Florida Mosq. Control Assoc. 9:24-28.
Revay, E. E., A. Junnila, R. D. Xue, D. L. Kline, U. R.
Bernier, V. D. Kravchenko, W. A. Qualls, N. Ghattas, and G. C. Müller 2013. Evaluation of commercial products for personal protection against
mosquitoes. Acta Trop. 125:226-230.
SAS Institute Inc. 2013. JMP®, Version 11. Cary, NC.
Snow, W. F. 1977. The height and direction of flight
of mosquitoes in West African savanna, in relation
to wind speed and direction. Bull. Entomol. Res.
67:271-279.
Tawatsin, A., S. D. Wratten, R. R. Scott, U. Thavara,
and Y. Techadamrongsin. 2001. Repellency of volatile oils from plants against three mosquito vectors. J. Med. Entomol. 26:76-82.
Xue, R. D., W. A. Qualls, M. A. Smith, M. K. Gaines,
J. H. Weaver, and M. Debboun. 2012. Field evaluation of the Off! Clip-on mosquito repellent
(metofluthrin) against Aedes albopictus and Aedes
taeniorhynchus (Diptera: Culicidae) in northeastern Florida. J. Med. Entomol. 49:652-655.
Yang, P., and Y. Ma. 2005. Repellent effect of plant
essential oils against Aedes albopictus. J. Vect. Ecol.
30:231-234.
BIFENTHRIN BARRIER SPRAY AGAINST AEDES
ALBOPICTUS AROUND AN URBAN CEMETERY,
ST. AUGUSTINE, FLORIDA
CHRISTOPHER BIBBS, CODI ANDERSON, MICHAEL L. SMITH,
AND RUI-DE XUE
ABSTRACT. Talstar® Professional (7.9%
bifenthrin, AI) was applied by a Stihl SR
420 gas powered backpack sprayer to the
perimeter vegetation of an 11 ha cemetery
in order to control the source of adult
Aedes albopictus into the surrounding residential neighborhoods. Prior to, and after
treatment, BG Sentinel traps baited with
BG-Lure were used to monitor adult abundance in the cemetery while egg production was monitored using black oviposition
cups. Surveillance continued for six weeks
post treatment with no additional treatments applied to the site. Adult Asian tiger populations were significantly reduced
in the cemetery one week after treatment
(97.75%) compared with areas with no
control measures. Population control declined, thereafter, to about 43% at four
weeks post treatment and was still significantly reduced despite a total rainfall of
12.7 cm (with a peak of 6.4 cm) during the
study period. The trend in egg reduction
from oviposition traps mirrored that of
the adult mosquito population. We found
that the barrier treatment was an effective method for controlling Ae. albopictus
where other methods, especially source reduction, have limited utility.
abundant and invasive anthropophilic mosquitoes commonly encountered in urban
environments (Lounibos et al. 2001). The
ubiquity of this species, especially when in
association with human environs, poses
significant problems as the result of its successful ability to colonize cryptic oviposition sites and rapidly produce large populations. Moreover, Ae. albopictus is a known
vector of multiple arboviruses, including
chikungunya (CHIKV), dengue, and the
recent Zika virus (Derraik and Slaney
2015, Ngoagouni et al. 2015, Wilson and
Chen 2015). In 2014, the Florida Department of Health confirmed locally acquired
cases of CHIKV from several counties.
Cemeteries provide a good example of
cryptic environments suitable for Ae. albopictus production and harborage. These
areas are often large sites with static waterholding containers such as flower urns built
into, or mounted on, head stones. This environment often contains extensive patches
of high density vegetation such as bamboo
thickets that can create small containers for
mosquito larvae when pruned back (Washburn and Hartmann 1992, Johnson and
Sukhdeo 2013). This situation can lead to
highly localized, but heavy, outbreaks as
adult Ae. albopictus host seek during the
day. Source reduction is often a principle
method in eliminating oviposition and larval production habitat for this species but
may be ineffective in some instances.
Therefore, the best approach for building a repertoire of effective management
strategies to reduce Ae. albopictus populations and/or the transmission risk of arbo-
Key Words. Aedes albopictus, barrier spray,
bifenthrin, cemetery, residential area
I. INTRODUCTION
The Asian tiger mosquito, Aedes albopictus (Skuse), is considered one of the most
76
Bibb et al.: Bifenthrin Barrier Spray Against Aedes albopictus Around An Urban Cemetery
viral infection is through integrated vector
management (IVM). One method, with decades of usage, is barrier spraying (Trout
et al. 2007, Qualls et al. 2013). The Anastasia Mosquito Control District (AMCD)
is familiar with this method and has used
it to successfully control adult mosquitoes
(Qualls 2013). Other techniques,such as
thermal fogging with adulticides, have
been reintroduced into AMCD operational programs for control of adult containerinhabiting mosquitoes in order to bolster
the options for IVM (Xue et al. 2013).
But just having the technologies available
for use is not enough. Chemical application methods, either singular or in combination, must be continually re-evaluated
for their applicability to contemporary
problems of vector abatement in order to
stay one step ahead of emergent disease
threats like CHIKV, dengue, and Zika viruses. However, any tactics for long-term
control of Ae. albopictus populations would
be welcomed.
As mentioned earlier, pesticides applied to vegetation as a residual barrier for
adult mosquito control can provide several
versatile approaches for mosquito management. A study by Cilek (2008) showed
that bifentrhin applied to the perimeter
vegetation of a public park provided effective (>70%) control against adult Ae. albopictus and Culex quinquefasciatus Say, for
six weeks, in residual leaf bioassays. Using
this approach we tested the efficacy of insecticide barrier treatments to reduce Ae.
albopictus populations in a cemetery surrounded by an urban residential network
of neighborhoods. These neighborhoods
shared a fence line perimeter with the
cemetery and have experienced considerable annoyance from host seeking Ae. albopictus originating from the latter area.
Test Site: A local 11 ha cemetery (latitude 29.893712, longitude -81.337268) in
St. Johns County was targeted as a major
source of Ae. albopictus production. This
area was surrounded on all sides by urban
77
residential housing with high human activity. The property was chosen for its abundant larval sites for which source reduction education has had little impact. Larval
developmental habitats in the cemetery
included dense bamboo thickets, flower
urns, exposed crypts, and ground maintenance storage areas overloaded with tire
piles. These habitats were routinely irrigated as part of normal ground maintenance. Mosquito landing rate counts were
conducted in the cemetery for three minutes in 18 locations to identify where the
majority of Ae. albopictus activity occurred.
From this information, four locations were
chosen for the study and monitored from
June 1 through July 25, 2015.
Surveillance. Two BG Sentinel™ traps
(BGS) baited with a standard Biogents®
BG-Lure (Biogents AG, Regensburg, Germany) were placed at least 50 m apart in
each location (Figure 1). Traps were operated continuously for 48 h per week for
two weeks prior to treatment. In addition,
five 500 ml black oviposition cups, lined
with velour oviposition paper, were placed
in the same four locations and allowed to
collect eggs for 48 h per week during the
same period. After treatment, both surveillance methods were continued in the same
frequency for six weeks. Rainfall was recorded weekly. Collections from BGS traps
and ovicup papers were transported to the
lab. Captured mosquitoes were identified
to species and eggs from ovipapers were
counted using a light dissecting scope.
Barrier treatment. The outer perimeter
vegetation and subsequent fence line of the
entire 11 ha site was treated with Talstar®
Professional (7.9% bifenthrin AI, Philadelphia, PA) at a dilution rate of 11.4-L water
to 88.7 ml bifenthrin. Treatment was applied
using a Stihl SR 420 gas powered backpack
sprayer (Andreas Stihl Ag & Co. KG, Waiblingen, Germany) at a 3.3 L flow rate while
walking the area at 6.4 kph. Application
height was at 3 m with a 60 m penetration
depth for a total treatment area of 6400 m2.
The pesticide application was carried out
by a certified State of Florida Public Health
Pest Control applicator.
78
Figure 1. Surveillance layout of the 11 ha cemetery used as a test site (latitude 29.893712, longitude -81.337268)
St. Johns County, FL. Each location contained two BG Sentinel traps (baited with BG-Lure) and five oviposition
cups.
Statistical analysis Percent population
reduction was calculated using a proportion derived by T/(T+P) where T = the
population for a given week of treatment
and P = the population for the two weeks
of pre-treatment sampling. Statistical software JMP® version 10.1 (SAS Institute,
Inc. 2012) was used to run a paired t-test
on mean adult and egg surveillance data to
determine differences in population level
from the treatment compared with pretreatment level per week for 6 weeks.
Ethics statement. Those volunteers who
participated in landing rate counts were
briefed on the study procedures and made
aware of the health risks involved by their
participation. Written informed consent
was received from volunteers prior to the
start of the study. Volunteer consent is documented under the AMCD IRB Protocol
#10.13.2005 and approved for use of human subjects by the Board of Commissioners presiding over the AMCD.
Aedes albopticus populations were significantly reduced by 97.75% during week
one; 65.64% during week two; 58.72% for
week 3; and 43.39% at week 4 (Figure 2).
Population levels at weeks 5 and 6, post
treatment, were no longer significantly different than pretreatment levels. Egg abundance after treatment was also significantly
reduced (t = 4.7154, df = 3, p < 0.0181) for
the first four weeks.
Despite such a large source site, at 11
ha, the perimeter barrier spray still provided successful control of the resident
Ae. albopictus population without additional adulticide or larvicidal treatment.
Previously, this area served as an important source of adult Asian tiger mosquito
for neighborhoods surrounding it. The
cemetery is a largely unmanaged property compounded by dense bamboo privacy screens along the perimeter. Addi-
Bibb et al.: Bifenthrin Barrier Spray Against Aedes albopictus Around An Urban Cemetery
79
Figure 2. Weekly mean precipitation and abundance of adult Aedes albopictus from BG Sentinel (BG AVG) and
mosquito egg oviposition cups (EGG AVG) in an 11 ha cemetery before (week 1-2) and after (week 3-8) treatment
of perimeter vegetation with Talstar Professional (bifenthrin). Vertical lines at each data point are standard errors.
tionally, tire piles and human generated
debris that included artificial containers
expanded the available sources. As stated
earlier, traditional, methods of source reduction and custodial education of cemetery employees on identifying and removing urban mosquito sources were met with
resistance. Moreover, relatively few roads
for truck access into the cemetery limited
the effectiveness, and increased the cost of
any larviciding or adulticiding effort often
requiring multiple visits over a season. Barrier spraying with Talstar Professional has
shown, in this case, to be the most effective
means of reducing large urban sources of
adult Ae. albopictus.
Appreciation is extended to the custodians of the cemetery site who collaborated with the test evaluation. The operational technicians of the Anastasia Mosquito
Control District are thanked for their collaboration during this large barrier mission. This is a research report only. The
AMCD does not endorse any commercial
mentioned in this report.
Cilek, J. E. 2008. Application of insecticides to vegetation as barriers against host-seeking mosquitoes. J.
Cilek, J. E. and C. F. Hallmon. 2008. Residual effectiveness of three pyrethroids on vegetation against
adult Aedes albopictus and Culex quinquefasciatus in
screened field cages. J. Am. Mosq. Control Assoc.
24:263-269.
Derraik, J. G. and D. Slaney. 2015. Notes on Zika virus—an emerging pathogen in the South Pacific.
Aust. N. Z. J. Public Health 39:5-7.
Johnson, B. J. and M. V. K. Sukhdeo. 2013. Successional mosquito dynamics in surrogate treehole
and ground-container habitats in the northeastern United States: where does Aedes albopictus fit
in? J. Vector Ecol. 38:168-174.
Lounibos, L. P., G. F. O’Meara, R. L. Escher, N.
Nishimura, M. Cutwa, T. Nelson, and S. A. Juliano. (2001). Testing predictions of displacement of
native Aedes by the invasive Asian tiger mosquito
Aedes albopictus in Florida, USA. Biological Invasions, 3:151-166.
Ngaogouni C., B. Kamgang, E. Nakoune, C. Paupy,
and M. Kazanji. 2015. Invasion of Aedes albopictus
(Diptera: Culicidae) into central Africa: what consequences for emerging diseases? Paras. Vectors 8:191.
Qualls, W. A., M. L. Smith, and R. D. Xue. 2013. Successful applications of barrier treatments using
bifenthrin against mosquitoes in St. Johns County,
80
Florida, from 2006-2009. Tech. Bull. Florida Mosq.
SAS Institute, Inc. 2012. JMP, Version 10.1. Cary NC.
Trout, R. T., G. C. Brown, M. F. Potter, and J. L. Hubbard. 2007. Efficacy of two pyrethroids insecticides
applied as barrier treatments for managing mosquito (Diptera: Culicidae) populations in suburban
residential properties. J. Med. Entomol. 44:470-477.
Washburn, J. O. and E. U. Hartmann. 1992. Could Aedes
albopictus (Diptera: Culicidae) become established
in California tree holes? J. Med. Entomol. 29:9951005.
Wilson, M. E. and L. H. Chen. 2015. Dengue: update in
epidemiology. Curr. Infect. Dis. Rep. 17:457.
Xue, R. D., J. H. Weaver, M. L. Smith, W. A. Qualls.
2013. Field evaluation of thermal fog application
of sumithrin products against adult mosquitoes in
northeast Florida. Tech. Bull. Florida Mosq. Control
Assoc. 9:38-41.
LABORATORY EVALUATIONS OF SEVEN INSECT
REPELLENTS AGAINST THE LONE STAR TICK
AMBLYOMMA AMERICANUM
JODI M. SCOTT, ALI FULCHER, JOHN M. HENLZER, AND RUI-DE XUE
ABSTRACT. In laboratory bioassays, BioUD,
Bio Block-Organic Outdoor, Bio BlockOrganic Pest Control, Bio Blocker-Organic
Insect Repellent, OFF! Botanicals Insect Repellent, OFF! Deep Woods Insect Repellent,
and Repel Insect Repellent Sportsman Gear
Smartinsect repellents were applied to poster paper and assessed by four volunteers for
efficacy against unfed nymphal Amblyomma
americanum ticks. Assessment times of 10 and
120 minutes post application were chosen
to mimic freshly applied repellent and suggested duration of effectiveness against ticks
as determined by the product manufacturers. Significant differences between repellents existed when both application periods
were compared but not between volunteers.
At 10 min post application, Bio Block Pest
Repel was the most effective product at repelling ticks (85%) and Repel the least effective
(30%). At 120 min, BiteBlocker-Insect Repel,
OFF! Deepwoods, and Bio Block-Organic
Outdoor provided ≥55% protection from
ticks. Efficacy at 120 min, when measured at
50% or greater repellency, did not reflect duration times listed on the BioUD (20%), and
OFF! Botanicals (30%) product labels.
mendations on how to avoid tick bites. In addition, AMCD’s field technicians frequently
encounter these ticks when conducting
fieldwork. Topical and barrier insect repellents offer an inexpensive mode of protection that imposes the fewest limitations on
human activities (Staub et al. 2002). With
the possible transmission of tick-borne
pathogens, such as Ehrlichia chaffeensis, E.
ewingii, E. muris, Southern Tick-Associated
Rash Illness (STARI), and Francisella tularensis (Mixson et al. 2006, CDC 2015), as
well as the growing concerns over DEET
(N,N-diethyl-3-methylbenzamide)
based
products on the market (such as, smell,
possible bioaccumulation, and damage to
plastics and some clothing materials) as reported by Katz et al. (2008) and Semmler et
al. (2011), alternative tick repellents should
be evaluated. Therefore, a laboratory study
was conducted to determine the repellent
effectiveness of several commercially available non-DEET products against lone star
ticks.
Insect repellents. Seven DEET and nonDEET commercially available products were
chosen to provide diversity of active ingredients that may be repellent to A. americanum
(Table 1). HOMS products were supplied
to AMCD for evaluation of efficacy by the
manufacturer, while other repellents were
purchased at local pharmacies or hardware
stores. HOMS Bio Block Organic Outdoor
and HOMS Bio-Block Organic Pest Control
are designed for premise treatment while
the rest of the products that we tested were
labeled for application to human skin.
Key Words: Insect repellents, DEET, ticks,
Amblyomma americanum
I. INTRODUCTION
Amblyomma americanum (L.), the lone
star tick, is a commonly encountered tick in
northeastern Florida. Annually, Anastasia
Mosquito Control District (AMCD) receives
multiple customer complaints about this
tick species and is often asked for recom81
2
1
SC Johnson & Sons
SC Johnson & Sons
WPC Brands, Inc.
HOMS. LLC
HOMS, LLC
Brand name formulations are subject to change; labels should always be read to ensure exact ingredients.
N,N-diethyl-3-methylbenzamide
Pump spray, Lotion
Aerosol Spray
No longer available
120 minutes
Not listed
480 minutes
OFF!-Botanicals® Insect Repellent
OFF!-Deep Woods, Insect Repellent
Repel®-Insect Repellent, Sportsman Gear Smart
Pump spray, Lotion, Travel pen
120 minutes
BiteBlocker®-Organic, Insect Repellent
Indoor-Monthly, Outdoor-Weekly Pump Spray
Bio Block® -Organic, Pest Control
120 minutes
Outdoor-Monthly
2-undecanone 7.75%
Soybean Oil 20%, Citric Acid 4%,
Cedar Oil 1%
Soybean oil 12%, Geranium oil 3%,
Caster oil 1%, Lemon Grass oil 0.5%
Soybean Oil 30%, Citric Acid 4%,
Cedar Oil 1%
p-Menthane-3,8-Diol 10%
DEET2 25%
Picardin 15%
HOMS, LLC
HOMS, LLC
BioUD®
Bio Block® -Organic Outdoor
Duration of protection
Brand Name
Active Ingredients1
Manufacturer
Table 1. Manufacturer, active ingredient (%), commercial product, formulation and manufacturer’s stated duration of protection.
Pump spray
Concentrate
Formulation
82
Human volunteers. Male (2) and female
(2) volunteers were utilized in these evaluations. Ages of the volunteers ranged from
24-55 years old and signed consent forms
under AMCD’s IRB protocol#10-13-2005 (as
approved by the AMCD Board of Commissioners for studies involving use of human
subjects).
Ticks. Non-blood fed, nymphal A. americanum were obtained from a laboratory colony
maintained by USDA/ARS, Tick and Biting Fly Research Unit, Knipling-Bushland
U.S. Livestock Insect Research Laboratory,
Kerrville, TX. New ticks were used by each
volunteer during each repellent evaluation.
After individual volunteers evaluated repellency, the ticks were collected by scotch tape
and disposed of.
Bioassays: The bioassay utilized a modification of human skin test evaluations
used by Semmler et al. (2011). White poster paper (1 meter x 1 meter) was used as
a substratum for all repellent applications.
Poster boards were taped onto individual
tables (183 cm x 76 cm x 74 cm) located
one meter from each other and designated
as either repellent treated poster boards or
untreated (control) posterboards. Three
different concentric circles were drawn
onto each poster paper: 15.5 cm outer
circle, 14.5 cm middle circle, and 5.5 cm
inner circle. Following label instructions
for application, repellents were painted inbetween the 15.5 cm and 14.5 cm circles
and allowed to dry for ten minutes. After
ten minutes, 5 ticks were placed with forceps onto the center of the 5.5 cm circle.
Volunteers sat in a chair at each table and
placed their arms to the left and right of
the outer 15.5 cm circle, ensuring no contact with repellent. Ticks were given 3 min
to locate volunteers. Successful location
was recorded when ticks crossed the 5.5 cm
center circle and came into contact with
14.5 cm middle circle. At the end of three
minutes, ticks were collected with scotch
tape, and the next volunteer evaluated
the same repellent using new ticks. Each
repellent was evaluated in this manner. A
secondary evaluation, 120 minutes after
initial application, was conducted which
Scott et al.: Laboratory Evaluations of Seven Insect Repellents Against The Lone Star Tick followed the previously outlined bioassay.
Ticks on the control poster board followed
the same bioassay procedure as the treated
ones.
Statistical analysis. The statistical software
JMP version 11.1.1 (SAS Institute Inc. 2013)
was used for data analysis. Statistical significance was determined through ANOVA
(P<0.05). Post hoc analysis was conducted
to separate the means of repellents using
Tukey HSD.
Our study focused on insect repellents
that offered a wide variety of active ingredients and evaluated their efficacy when mimicking freshly applied at 10 min and at 120
min post application to mimic the label duration of effectiveness listed by most of the
manufacturers. Significant differences existed between products but not between volunteers, at both post application time periods
(F = 5.1137, df10,21 = 31, P = 0.0008, and F =
4.0310, df10,21 = 31, P = 0.0034, respectively)
(Table 2). At 10 min post application, Bio
Block Pest Repel was the most effective product at repelling ticks (85%) while Repel was
the least effective (30%). At 120 min, Bite
Blocker Insect Repel, OFF! Deep woods, and
Bio Block-Organic Outdoor provided ≥55%
protection from ticks and was consistent with
their respective labeled protection rates as
listed in the EPA Insect Repellents Use and
Table 2. Mean percent repellency (SD) of seven insect repellents against non-blood fed Amblyomma
americanum nymphs at 10 min and 120 min post application.
None
BioUD
Repel
BioBlock-Pest Control
OFF! Deep Woods
BioBlock-Outdoor
OFF Botanicals
BiteBlocker-Insect Repel
10 min1
0 (0)c
50 (35)abc
30 (25)bc
85 (19)a
80 (28)ab
70 (26)ab
70 (26)ab
80 (16)ab
Effectiveness guidelines (EPA 2015). At 120
min, the efficacy of BioUD (20%), and OFF!
Botanicals (30%), measured at 50% or greater repellency, did not reflect labeled duration
times listed on their labels. We believe these
studies have provided several options, for the
general public and our field technicians, on
what commercially repellents are efficacious
and available for protection against nymphal
host seeking A. americanum.
III. RESULTS
Repellents
83
20 min1
5 (10)b
20 (16)ab
50 (26)ab
50 (12)ab
65 (25)a
55 (25)a
30 (35)ab
65 (19)a
1
Means within each time-period, between products, followed by a different letter are statistically different (Tukey
HSD).
All volunteers gave informed consent regarding the possible risks of acquiring tickborne diseases during the laboratory trials.
This is a research report only, use or mention of trade names does not constitute an
official endorsement by Anastasia Mosquito
Control District.
V. REFERENCES CITED
Achee, N. L., M. J. Bangs, R. Farlow, G. F. Killeen, S.
Lindsay, J .G. Logan, S. J. Moore, M. Rowland, K.
Sweeney, S. J. Torr L. J. Zwiebel, and J. P. Grieco.
2012. Spatial repellents: from discovery and development to evidence-based validation. Malaria Journal 11:164 doi: 10.1186/1475-2875-11-164.
Carroll, J. F., J. P Benante, M. Kramer, K. J. Lohmeyer,
and K. Lawrence. 2010. Formulations of deet, picaridin, and IR3535 applied to skin repel nymphs of the
Lone Star tick (Acari: Ixodidae). J. Med. Entomol.
47:699-704.
2015. Tickborne disease of the U.S. http://www.cdc.
gov/ticks/diseases/.[Accessed 8 July 2015].
Childs, J. E. and C. D. Paddock. 2003. The ascendancy
of Amblyomma americanum as a vector of pathogens
affecting humans in the United States. Annu. Rev.
Entomol. 48:307-377.
EPA (Environmental Protection Agency). 2015. Insect
repellents: use and effectiveness. http://cfpub.epa.
gov/oppref/insect/search_results.cfm?Rangetime
=&hidSelected=3&ProductName=&Ingredient=nul
l&Company=null&Registration=&Submit=Search#.
[Accessed 1 July 2015].
Gratz, N. G. 1999. Emerging and resurging vector-borne
diseases. Annu. Rev. Entomol. 44:51-75.
Katz, T. M., J. H. Miller, and A. A. Herbert. 2008. Insect
repellents: historical perspectives and new developments. J. Am. Acad. Dermatol. 58:865-871.
Mixson, T. R., S. R. Campbell, J. S. Gill, H. S. Ginsberg,
M. V. Reichard, T. L. Schulze, and G. A. Dasch. 2006.
Prevalence of Ehrlichia, Borrelia, and Rickettsial agents
in Amblyomma americanum (Acari: Ixodidae) collected from nine states. J. Med. Entomol. 43:1261-1268.
Moody, R. P. 1989. The safety of diethyltoluamide insect
repellents. J. Am. Mosq. Control Assoc. 262:28-29.
84
Osimitz, T. G. and R. H. Grothaus. 1995. The present
safety assessment of DEET. J.Am. Mosq. Control Assoc. 11:274-278.
SAS Institute Inc. 2013. JMP statistical software. Version
11.1.1 Cary, NC.
Semmler, M., F. Abdel-Ghaffer, K. A. S. Al-Rasheid, and
H. Mehlhorn. 2011. Comparison of the tick repellent efficacy of chemical and biological products
origination from Europe and the USA. Parasitol.
Res. 108:899-904.
Staub, D., M. Debrunner, L. Amsler, and R. Steffen.
2002. Effectiveness of repellent containing DEET
and EBAAP for preventing tick bites. Wild. Environ
Med. 13:12-20.
Xue, R. D., A. Ali, and J. F. Day. 2007. Commercially
available insect repellents and criteria for their use.
In: Insect Repellents, Principles, Methods, and Uses, ed.
by M. Debboun, S.P. Frances, and D. Strickman.
CRC Press. Boca Raton, FL, USA, Chapter 25, 405415 pp.
LABORATORY AND FIELD EVALUATION OF OFF! CLIPON MOSQUITO REPELLENT DEVICE CONTAINING
METOFLUTHRIN AGAINST THE LONE STAR TICK,
AMBLYOMMA AMERICANUM (ACARI: IXODIDAE)
RUI-DE XUE¹, JODI M. SCOTT¹, ALICIE FULCHER¹,
WHITNEY A. QUALLS¹, JOHN M. HENLZER¹,², MARCIA K. GAINES¹,
JAMES H.R. WEAVER¹, AND MUSTAPHA DEBBOUN³
1
2
3
Guana Tolomato Matanzas Research Reserve, Environmental Education
Center, Florida Department of Environmental Protection
505 Guana River Road, Ponte Vedra Beach, FL 32082
Mosquito Control Division, Harris County Public Health & Environmental
Services, Houston, TX 78234
ABSTRACT. Choice tests were conducted in the laboratory to determine the
repellency of the pyrethroid insecticide
metofluthrin (Off!® Clip-on™ Mosquito
Repellent, AI 31.2%) to lone star ticks,
Amblyomma americanum. In this study, the
device’s fan unit was disabled to allow evaluation of the active ingredient as a passive
spatial repellent. We found that 57% of
the ticks were significantly repelled from
hands that held the Off! Clip-on refill,
compared with 27% of volunteer hands
without it (control). Field trials conducted
in northeastern Florida with volunteers either sitting or walking while wearing the
repellent device in areas infested with A.
americanum were also evaluated. Seated
volunteers, with the device attached to
their waist, with the fan running repelled
89% of host-seeking ticks when compared
with persons similarly placed without the
fan on. In a second field trial, volunteers
that walked slowly through the naturally
infested area with the repellent fan running provided 28% protection from questing ticks compared with similar volunteers
without the fan on. Our overall results
demonstrated that fewer numbers of lone
star ticks were found on persons with the
Off! Clip-on Mosquito Repellent device
operating compared with those wearing
the device in the off position, regardless
of sitting or walking slowly through a tick
infested area.
Key Words. Amblyomma americanum, tick
repellent, metofluthrin, personal protection, Off Clip-on Mosquito Repellent
I. INTRODUCTION
Tick-borne diseases in humans and
animals are a growing world-wide concern
and problem (Gratz 1999). A common tick
in the southeastern US is the lone star tick,
Amblyomma americanum (L.) and has been
reported to be a potential vector of human monocytic ehrlichiosis as well as several other emerging tick-borne pathogens
(Childs and Paddock 2003). Anastasia
Mosquito Control District clientele, and
our field staff, have experienced numerous
encounters with this pest tick species after
visiting some of the state parks in St. Johns
County, Florida (Xue, pers. comm.). Tick
control using an acaricide is very difficult
in Florida state parks because these areas
are considered conservation lands where
pesticide applications are prohibited.
85
86
Therefore, personal protective measures
rely on topical application of repellents as
the most effective protection against host
seeking ticks (Carroll et al. 2010).
Most common commercially available
repellent products contain different concentrations of N, N-diethyl-3-methylbenzamide (DEET) as the active ingredient
(Xue et al. 2007). Due to concerns about
DEET’s potential toxicity and safety (Osimitz and Grothaus 1995), including possible
health risks associated with repeated use of
high concentrations (Moody 1989), spatial
repellents have been suggested as alternatives. Theoretically, an effective spatial
repellent should provide an envelope of
protection that prevents contact between
host and pest while preventing pathogen
transmission if disease vectors are present
(Achee et al. 2012). Most arthropod spatial
repellent products release an active ingredient as a vapor or smoke. Both dispersal
methods often use a heat source, or fan,
to produce the protection zone. Metofluthrin, a vapor-active pyrethroid, has been
identified as an effective spatial mosquito
repellent (Ujihara et al. 2004, Kawada et
al. 2005). Several novel methods of applying spatial repellents against mosquitoes
have included impregnated paper (Argueta et al. 2004; Lucas et al. 2007) and plastic
strips (Kawada et al. 2008). Recently, Off!®
Clip-on™ Mosquito Repellent was introduced as a new commercially-available spatial repellent device; this product has been
tested against mosquitoes and provided effective protection in northeastern Florida
(Xue et al. 2012). To date, Off! Clip-on has
not been evaluated (nor its active ingredient) for personal protection against ticks.
Here we report the results of laboratory
and field evaluations conducted in northeastern Florida, to determine the repellency of Off! Clip-on Mosquito Repellent
against host-seeking adult and nymphal A.
americanum.
Spatial repellent device. Off! Clip-on Mosquito Repellent devices were purchased
from a local convenience store. This device consists of a plastic dispenser containing a replaceable cartridge with metofluthrin (AI 31.2%). This product is designed
to operate by turning on a fan powered
by two 1.5 volt AA alkaline batteries to assist in the release of the active ingredient
into the air surrounding the individual.
According to the label, the device can be
worn by adults or children for protection
against mosquitoes for up to 12 h of operation.
Laboratory test. Static, non-fan-mediated
activity of metofluthrin was evaluated in
laboratory cage bioassays. Each cage was
constructed from a plastic Sterilite® box
(13 × 13 × 21.5 cm; L × W × H, Sterilite,
Townsend, MA) and served as an open
airflow chamber (OAC) for choice evaluations (Figure 1). A clear vinyl tube (3.5 cm
outer diameter × 30 cm) was connected to
the OAC through a 3.5 cm diameter hole
on the side. The tube was then connected to a 1,000 ml polystyrene disposable
storage bottle (Corning®, Corning, NY)
through a 3.5 cm diameter hole in the center of the bottle lid. The base of the bottle
had been removed to allow insertion of a
volunteer’s hand.
Laboratory-reared (non-blood fed)
adult A. americanum were used in the study
from a colony maintained by the USDA/
ARS, Tick and Biting Fly Research Unit,
Knipling-Bushland U.S. Livestock Insect
Research Laboratory, Kerrville, TX. For
each test, five ticks were placed into the
vinyl tube, using feather tip forceps, and
secured with fabric mesh held in place by
rubber bands to ensure that no ticks escaped the tube. Ticks were allowed 2 min
to acclimate in the tube before it was attached to the OAC and polystyrene bottle.
A volunteer then placed one hand into
the end of the polystyrene bottle and ticks
were given 5 min to move to their final
position. After five minutes, ticks located
between the hand, on the mesh, and up
to 15 cm from the hand were counted as
“host seeking”. Ticks were given a 2-min
rest period then the assay repeated but
with the volunteer holding an OFF! Clip-
Xue et al.: Laboratory and Field Evaluation of Off! Clip-On Mosquito Repellent Device 87
Figure 1. Choice tube laboratory bioassay for evaluating adult female Amblyomma americanum response to a human hand with and without Off! Clip-on Mosquito Repellent cartridge containing metofluthrin.
on refill while placing their hand in the
polystyrene chamber. Ticks were allowed
10 min to locate the volunteer’s hand. After 10 min, ticks located on the mesh next
to the hand, and within 15 cm of the hand,
were counted as attracted to the host. For
all tests, the percentage responding was
calculated. Three volunteers conducted
three sessions each providing three replications with an N = 9. New ticks were used
for each replication.
Field test. Field trials were conducted
at Guana State Park (permit# 2012-3-12)
in South Ponte Vedra, Florida (30° 07”
56.37”N and 81° 22” 40.44 W). This land
parcel is a conservation area where large
populations of lone star ticks occur. Six
human volunteers (four males and two
females) who each signed a consent form
under protocol No. 445-96, as approved
by the University of Florida, Institutional
Review Board (IRB-01) participated in the
trials. Volunteers wore white clothing that
fully covered their body from head to toe.
Each person had one Off! Clip-on device
attached to their waist and sat on a chair
15 m away from the nearest volunteer.
Three volunteers served as treatments,
with their device fan turned on for 30 min.
During the same time period, the three
remaining volunteers had an Off! Clip-on
device attached to their waist but without
the fan operating (referred to as “static
device”) and served as controls. All stages
of ticks that climbed upon the volunteers
clothing were collected by scotch tape by
the end of the observation period. All volunteers then relocated to a new testing
site in the same area and switched treatments, (i.e. previous control volunteers
switched to serve as the treatment and the
previous treated volunteers now served as
the control). This scenario was repeated 6
times on one day during June 2012. Meteorological data (wind direction, speed, air
temperature, and relative humidity) were
collected hourly with a small hand-held
digital meter (Skywatch Xplorer, N Tech,
USA, Holmen, WI).
In the second field trial at the same
state park, six volunteers (3 females and
3 males) participated. Three volunteers
wore devices, attached to their waist, without operating the fan while the remaining
88
three volunteers wore devices where the
fan was operated. Volunteers wore the previously mentioned white suits from the first
trial and walked at a pace covering 30 m
for 15 min. Each volunteer was positioned
15 m from the next volunteer. Ticks were
removed from volunteers by the end of
each time period. After the 15 min collection period, volunteers relocated to a new
test site and either switched on or off their
fans, to provide a comparison to their previous treatment assignment. These field
trials were repeated six times on one day
from 0800 to 1500 during late May, 2012.
Data analyses. Mean percent response
data from the laboratory study were subjected to an analysis of variance using JMP
version 11 (SAS Institute, Inc. 2013) on
non-transformed data, to assess statistical
significance between controls and treatments on pooled data across all volunteers. Tukeys HSD was then conducted on
this dataset to determine if differences existed (P<0.05). Field trial repellency data
was evaluated in a split-plot design. The
data for walking and sitting were analyzed
separately. Mean tick counts were transformed using square root (x + 1) prior to
analyses and Student’s t-test performed to
determine differences (P<0.05).
III. RESULTS
Laboratory test. There was statistical significance between the treatment and the
control (F1,16 = 9.7826, P = 0.0065). Fiftyseven percent of ticks were repelled from
the volunteer holding the clip on refill,
while 27% ticks were repelled from the volunteer without holding the clip on refill.
There was no statistical difference between
volunteers in these trials (F2,15 = 0.3439, p
= 0.7144).
Field test. Only one tick species, A. americanum, was collected in the field trials. A
total of 98 ticks were collected including
6 females, 3 males, and 89 nymphs. At
30 min, a mean of 3 ticks were recovered
from seated volunteers holding the clip
on devices. During the same time period,
2 ticks were recovered from only one of
the six volunteers with operating devices
for a mean of 0.33 ticks. In addition, 2-6
ticks were noticed around the feet of the
three sitting volunteers while the fan was
on but they did not climb onto either person. During this part of the field study,
89% protection was provided by the clipon device operating against questing ticks
for volunteers who were siting. Wind speed
during the trial was recorded at 0-9 km/h
with air temperature ranging between 25
and 28° C (70% RH).
At the beginning of the walking field trial, numerous ticks were initially observed
on the feet and legs of all volunteers.
However, those with the repellent device
turned on noticed that those ticks either
left or dropped off each person while the
device was operated for a few minutes. A
mean of 7.3 ticks per person per 15 min
were observed on volunteers with the
device off and 5 ticks per person per 15
min on persons with an operating device.
The percent protection from questing A.
americanum afforded to walking individuals by operating OFF! Clip-on device was
28 % compared with persons with a nonoperational device. In this field trial, there
were no significant differences in the numbers of ticks recovered between volunteers
when using, or not using, the fan component. During this study wind speed was recorded at 0-3.6 km/h with air temperature
ranging from 26–28° C (70% RH) in the
morning of the second field trial.
IV. DISCUSSION
Host-seeking ticks can approach a host
by walking or they may sit and wait for
the host to make contact, a process called
questing. Usually questing is the primary
host seeking behavior for ticks rather than
directly approaching a host. Therefore,
walking is an ideal method to evaluate tick
repellents for those species that primarily
utilize questing to acquire hosts. Because
few ticks actively move toward a host, and
most ticks walk/approach a host very slowly, sitting for 15 min periods may be a less
Xue et al.: Laboratory and Field Evaluation of Off! Clip-On Mosquito Repellent Device effective method for a repellent evaluation
against these organisms.
We found that operating an Off! Clipon mosquito repellent device containing
metofluthrin proved to be more effective
(89%) for protecting inactive (sitting)
hosts from climbing ticks than it was for
hosts that were actively walking (28%).
This effect was most likely influenced by
entering tick harborage areas and the
tick’s ability for host location through its
questing behavior. To provide more effective protection for individuals walking
in tick infested habitats, the Off! Clip-on
device needs to be improved by possibly
increasing the concentration of metofluthrin, fan speed, and/or wearing the device
on the lower part of legs rather than on
the waist.
Although the Off! Clip-On Mosquito
Repellent was designed to primarily repel
mosquitoes, we observed that a certain
amount of effectiveness against crawling A.
americanum on clothing can be achieved.
Moreover, we found that the Off! device
provided effective protection from crawling ticks especially when people sit outdoors.
The Off! Clip-on Mosquito Repellent
system is easy to operate and can be attached on a belt, purse, or placed on a flat
surface for protection against host-seeking
arthropods (Xue et al. 2012). The active
ingredient, metofluthrin, can last for up to
12 h (for mosquitoes) and used multiple
times but must be refilled within 14 days
after opening according to the manufacturer. However, the actual longevity of the
active ingredient in this device (measured
in hours or days) as personal protection
against host-seeking ticks has not been
evaluated.
V. ACKNOWLEDGEMENTS
We thank the volunteers who participated in the partial or whole field trials.
All volunteers gave informed consent regarding the possible risks of getting tickborne diseases during the field trials. This
is a research report only. Use or mention
89
of trade names does not constitute an official endorsement by Anastasia Mosquito
Control District.
Achee, N. L., M. J. Bangs, R. Farlow, G. F. Killeen, S.
Lindsay, J. G. Logan, S. J. Moore, M. Rowland, K.
Sweeney, S. J. Torr, and J. P. Grieco. 2012. Spatial
repellents: from discovery and development to
evidence-based validation. Malaria Journal 11:164
doi: 10.1186/1475-2875-11-164.
Argueta, T. B. O., H. Kawada, and M. Takagi. 2004.
Spatial repellency of metofluthrin-impregnated
multilayer paper against Aedes albopictus under
outdoor conditions, Nagasaki, Japan. Med. Entomol. Zool. 55:211-216.
Carroll, J. F., J. P. Benante, M. Kramer, K. J. Lohmeyer, and K. Lawrence. 2010. Formulations of
deet, picaridin, and IR3535 applied to skin repel
nymphs of the lone star tick (Acari: Ixodidae). J.
Childs, J. E. and C. D. Paddock. 2003. The ascendancy of Amblyomma americanum as a vector of pathogens affecting human in the United States. Ann.
Rev. Entomol. 48:307-377.
Gratz, N. G. 1999. Emerging and resurging vectorborne diseases. Ann. Rev. Entomol. 44:1-75.
Kawada, H., E. A. Temu, J. N. Minjas, O. Matsumoto,
T. Iwasaki, and M. Takagi. 2008. Field evaluation
of spatial repellency of metofluthrin-impregnated
plastic strips against Anopheles gambiae complex in
Bagamoyo Coastal Tanzania. J. Am. Mosq. Control
Assoc. 24:404-409.
Kawada, H., Y. Maekawa, and M. Takagi. 2005. Field
trial on the spatial repellency of metofluthrin-impregnated plastic strips for mosquitoes in shelters
without walls (beruga) in Lombok, Indonesia. J.
Vect. Ecol. 30:181-185.
Kawada, H., Y. Maekawa, Y. Tsuda, and M. Takagi.
2004. Trial of spatial repellency of metofluthrinimpregnated paper strip against Anopheles and Culex in shelters without walls in Lombok, Indonesia.
J. Am. Mosq. Control Assoc. 20:434-437.
Lucas, J. R., Y. Shono, T. Iwasaki, T. Ishiwatari, N.
Spero, and G. B. Benzon. 2007. U.S. Laboratory
and field trials of metofluthrin (SumiOne) emanators for reducing mosquito biting outdoors. J.
Moody, R. P. 1989. The safety of diethyltoluamide
insect repellents. J. Am. Mosq. Control Assoc.
262:28-29.
Osimitz, T. G. and R. H. Grothaus. 1995. The present
safety assessment of DEET. Mosq. Control Assoc.
11:274-278.
SAS Institute, Inc. 2013. JMP. Version 11. Cary, NC.
Ujihara, K., T. Mori, T. Iwasaki, M. Sugano, Y. Shono, and N. Matsuo. 2004. Metofluthrin: a potent
new synthetic pyrethroid with high vapor activity
against mosquitoes. Biosci. Biotechnol. Biochem.
68:170-174.
Xue, R. D., A. Ali, and J. F. Day. 2007. Commercially
available insect repellents and criteria for their
use. In: Insect Repellents, Principles, Methods, and
Uses. M. Debboun, S. P. Frances, and D. Strickman
90
(eds). CRC Press, Boca Raton, FL, USA, chapter
25, pp. 405-415.
Xue, R. D., W. A, Qualls, M. L. Smith, M. K. Gaines,
J. H. Weaver, and M. Debboun. 2012. Field evaluation of the Off! Clip-on mosquito repellent
(metofluthrin) against Aedes albopictus and Aedes
taeniorhynchus (Diptera: Culicidae) in northeastern Florida. J. Med. Entomol. 49:652-655.
Zollner, G. and L. Orshan. 2011. Evaluation of a
metofluthrin fan vaporizer device against phlebotomine sand flies (Diptera: Psychodidae) in a
cutaneous leishmaniasis focus in the Judean Desert, Israel. J. Vect. Ecol. 36(suppl.): s157-165.
Operational Note
EVALUATION OF TALENT UV LIGHT TRAPS COMPARED
WITH CDC LIGHT TRAPS, WITH OR WITHOUT DRY ICE,
TO COLLECT FRESH AND SALT WATER MOSQUITOES IN
NORTHEAST FLORIDA
MICHAEL L. SMITH, WHTINEY A. QUALLS¹, AND RUI-DE XUE
Miller School of Medicine, University of Miami
Miami, FL 33124
1
ABSTRACT. The Talent UV light trap was
compared with the CDC light trap, baited
with or without dry ice, to determine the
best trapping method to collect saltmarsh
and freshwater mosquitoes on Anastasia Island and southern St. Johns County, Florida.
Seven species of mosquitoes were collected
from a saltmarsh on Anastasia Island, of
which Aedes taeniorhynchus was the major
species (90%). Seven species of mosquitoes
were collected from freshwater habitats in
southern St. Johns County, where Culex quinquefasciatus was the dominate species (81%).
Overall, the CDC trap collected significantly
more mosquitoes than the UV trap. For both
traps, the addition of dry ice considerably increased mosquito abundance in collections.
trap has been developed for the surveillance
of adult mosquitoes but has not been evaluated for the collection of saltmarsh mosquitoes. The objective of the study was to
compare the Talent UV light trap with the
gold standard CDC light trap in order to determine the best trapping method to collect
saltmarsh and freshwater mosquitoes in Anastasia Mosquito Control District.
Six Talent UV traps (model MSTRS),
modified from a standard CDC light trap,
were supplied by J. W. Zhu (Iowa State University, Ames, Iowa) and compared with 6
standard CDC light traps (John W. Hock
Company, Gainesville, FL). The Talent UV
traps were modified by adding a 190 mm
(L) × 63 mm (W) × 32 mm (H) UV light bar
that produced a light wavelength of 325 nm.
Both traps were placed in a random complete block design separated by 30 meters in
a residential community on Anastasia Island,
St. Augustine, FL for the collection of saltmarsh mosquitoes. The same experimental
design was used to evaluate the collection of
freshwater mosquitoes in a companion study
in the southern part of St. Johns County. All
traps were operated 4 nights per week over
a period of three weeks. Each trap was randomly assigned a location during each trap
night. During the first field trial, Talent traps
without dry ice were compared with dry icebaited CDC light traps (Table 1). During the
second field trial, Talent traps were baited
Key Words. UV light trap, CDC light trap,
dry Ice, Aedes taeniorhynchus, Culex quinquefasciatus
Insect suction traps, with and without attractants, are major tools used for surveillance
of adult mosquito populations (Kline 2006).
Evaluation of new traps and attractants for
use within operational mosquito surveillance
programs continue to be an active area of research in the field of medical entomology.
Mosquitoes perceive ultraviolet light (UV)
(Allan 1994) and traps have been evaluated
that use this wavelength for illumination (e.g.
Allan 1994, Li et al. 2015). The Talent UV
91
92
Table 1. Mean number (±SE) of mosquitoes collected by Talent UV traps and CDC traps baited with dry ice or
without dry ice from saltmarsh and freshwater habitats, northeastern Florida1.
Saltmarsh
Freshwater
Trap type
Dry Ice
No Dry Ice
Dry Ice
No Dry Ice
CDC Trap
Talent UV Trap
73 ± 0.5
57 ± 3.0
91 ± 4.3
25 ± 8.3
46 ± 13.0
19 ± 6.3
38 ± 12.7
32 ± 4.3
1
Saltmarsh habitat mosquito collections consisted of 90% Aedes taeniorhynchus: χ² = 16.999, df = 2, P < 0.01; freshwater collections consisted of 81% Culex quinquefasciatus: χ² = 23.491, df = 2, P < 0.01.
with dry ice while the CDC traps were operated without dry ice. In the third field trial,
Talent traps and CDC traps were each operated without dry ice. One gallon Igloo containers purchased from local suppliers were
used to dispense the dry ice from one pound
blocks. Traps were placed 1.5 meters above
ground using a Shepard’s hook.
Trap data collected from saltmarsh and
freshwater areas were separately analyzed
using non-parametric Kruskal-Wallis χ² tests
for each trap type and attractant combination. Differences were determined at P <
0.05.
A total of seven mosquito species were
collected from the saltmarsh habitat. The
species composition (and percent abundance in total collection) were: Aedes taeniorhynchus (Weidemann) (90%), Anopheles crucians Weidemann (7%), and Culex nigripalpus
Theobald (2%) with Ae. sollicitans (Walker),
Cx. erraticus (Dyar and Knab), Ae. triseriatus
(Say), and Masonia dyari Belkin, Hinemann,
and Page making up the remainder of the
collection. Seven species of mosquitoes were
collected from freshwater habitats including Cx. quinquinfasciatus Say (81%), An. crucians (12%), Ae. atlanticus (Dyar and Knab)
(3%), and Psorophora ferox (von Humbolt)
(3%) with the remainder of the collection
represented by Ae. triseriatus, Culiseta melanura (Coquillett), and Coquillettidia perturbans
(Walker).
In the freshwater habitat, mean abundance of mosquitoes (trap/night) from
both traps were not significantly different
in the absence of dry ice (Table 1). But unbaited CDC light traps collected significantly
more mosquitoes than unbaited Talent traps
in the saltmarsh habitat. We also found that
the mean number of mosquitoes from CDC
traps baited with dry ice were significantly
more than similar baited Talent traps. Regardless of what habitat they were collected
in.
In our study, both light traps collected
mosquitoes equally without the addition
of dry ice from the freshwater habitat but
this relationship changed considerably (i.e.
2-fold increase) when dry ice was added. It
is well known that carbon dioxide is an attractant that considerably increases adult
mosquito abundance in CDC light trap collections (Newhouse et al 1966, Kline 2006).
We also found that the CDC light trap outperformed the Talent trap in the saltmarsh
habitat, regardless if both traps were baited
with dry ice. These results are similar to what
Li et al. (2015) found in Elkton, St. Johns
County, FL. In summary, the CDC light trap
currently remains the best adult mosquito
surveillance trap available for the species we
collected in our study.
We thank Catherine Lippi for assisting in
data analysis. This is a research report only
and does not reflect the endorsement of the
Anastasia Mosquito Control District for any
of the products mentioned in this study.
REFERENCES CITED
Allan, S. A. 1994. Physics of mosquito vision-an overview. J. Am. Mosq. Control Assoc. 10:2661-2671.
Kline, D. L. 2006. Mosquito population surveillance
techniques. Tech. Bull. Florida Mosq. Control Assoc.
7:2-8.
Li, C. X., M. L. Smith, A. Fulcher, P. E. Kaufman, T. Y.
Zhao, and R. D. Xue. 2015. Field evaluation of three
new mosquito light traps against two standard light
traps to collect mosquitoes (Diptera: Culicidae) and
non-target insects in northeast Florida. Florida Entomol. 98:114-117.
Newhouse, V. F., R. W. Chamberlain, J. G. Johnson, and
W. D. Sudia. 1966. Use of dry ice to increase mosquito catches of the CDC miniature light trap. Mosq.
News 26:30-35.
Operational Note
EFFECTS OF LEAF WASHING ON THE PERSISTENCE OF
A SUGAR BAIT-PYRIPROXYFEN MIXTURE TO CONTROL
LARVAL AEDES ALBOPICTUS
JODI M. SCOTT¹, ALI FULCHER¹, WHITNEY A. QUALLS²,
GUNTER C. MULLER³, AND RUI-DE XUE¹
1
2
Miller School of Medicine, University of Miami
Miami, FL 33124
Department of Microbiology and Molecular Genetics
IMRIC, Kuvin Centre for the Study of Infectious and Tropical Diseases
Faculty of Medicine, Hebrew University
Jerusalem, Israel 91120
3
ABSTRACT. The effect of adding the insect
growth regulator, pyriproxyfen, to an attractive sugar bait (ASB) was evaluated in semifield bioassays against larval Aedes albopictus.
Treatments were applied to croton “Petra”
potted plants while control plants were
sprayed with ASB only. Once a week, for six
weeks, plants were “washed” with water to
simulate rainfall and the runoff collected for
adult emergence inhibition bioassays. Adult
inhibition in the ASB/pyriproxyfen treatment was about 90% for the first two weeks
of the study then gradually decreased from
about 80% in week three to about 30% at
week six.
this technique has been proven to be safe for
non-target insects when applied to non-flowering plants or incorporated in bait stations
(Qualls et al. 2012, Revay et al. 2013).
However, rainfall currently limits the effectiveness of ATSBs as barrier treatments
against adult mosquitoes. In order to turn
this limitation into an advantage, we report here on the addition of pyriproxyfen
to a commercial attractive sugar bait (ASB)
formulation to order to control Aedes albopictus (Skuse) larvae. Pyriproxyfen, by itself,
is an effective larvicide that inhibits adult
emergence according to laboratory bioassays reported by Ali et al. (1995). This active
ingredient has also provided effective larval
control of Ae. albopictus when applied by
truck-mounted equipment ULV equipment
in northeastern Florida (Scott et al. 2013 and
Doud et al. 2014). A recent laboratory study
by Fulcher et al. (2014) reported effective
control of adult and larval mosquitoes from
plants sprayed with an ASB and pyriproxyfen
mixture. However, it was unclear whether
natural precipitation on ASB pyriproxyfentreated plants would affect the efficacy and
persistence of Ae. albopictus larval control.
Therefore, the objective of this study was to
Key Words. Aedes albopictus, pyriproxyfen, attractive sugar bait, larvicide, croton “Petra”
Previous studies have shown that attractive toxic sugar baits (ATSB) sprayed on
plants will kill adult Aedes, Anopheles, and
Culex species (Xue et al. 2006, 2011, 2013,
Muller et al. 2010, Naranjo et al. 2013, Revay
et al. 2013, Hossain et al. 2014). Attractive
toxic sugar bait is a highly effective method
of controlling adult mosquitoes by exploiting their sugar feeding behaviors. Moreover,
93
94
determine the impact of leaf washing on efficacy and persistence of ASB/pyriproxyfen
applications on plants for larval Ae. albopictus
in a semi-field trial.
Eleven, 25 cm dia. croton, Codiaeum variegatum “Petra” (L.) Rumph ex. A. Juss, plants
were purchased from Home Depot, St. Augustine, FL. Plants were washed off and allowed to dry before application to the leaves.
A commercially available attractive sugar
bait concentrate was mixed with water 1:4
ratio (Westham Innovations LTD, Tel Aviv,
Israel) to which 1% NyGuard® ED (10%
pyriproxyfen AI, provided by Navy Entomology Center of Excellence) was then added to
the bait solution. The mixture was applied
to nine plants using a 5.6 L pump-up sprayer
(ACE half-gallon polypropylene sprayer) in
sweeping motions over the plants until complete coverage occurred but not to the point
of runoff. This technique simulated barrier
applications used in field environments. Two
control plants were sprayed with ASB only
in the same manner as treated plants. Only
one ATSB (and ASB for control) application
was made to plants during the study. After
a 24 hour drying period, 200ml of reverse
osmosis water, (GE SmartWater™ Reverse
Osmosis Filtration System, Fairfield, CN)
was applied by a handheld sprayer over the
surfaces of the leaves while clock-wise rotating the plants to simulate rainfall. The runoff was collected into an 11.4-L (40 cm × 31.5
cm × 15.2 cm) plastic dish pan (Sterilite Corporation, Townsend, MA) and transferred to
946 ml all-purpose-calibrated (APC) plastic
containers (Ace Hardware, St. Augustine,
FL). Plant washes were conducted once a
week for six weeks.
Aedes albopictus larvae, maintained in the
Anastasia Mosquito Control District, were
used in bioassays. This insecticide-susceptible laboratory colony was originally obtained from the USDA, Center for Medical,
Agricultural, and Veterinary Entomology,
Gainesville, Florida. Mosquito larvae were
reared at 27° C; 80% RH; 12:12 L:D photoperiod and used the procedures described
by Gerberg et al (1994). Ten late third to
early fourth instars were placed into each
APC plastic container containing runoff wa-
ter and maintained in the insectary. Larvae
were fed every three days with 0.03 g ground
dog biscuits. Emergence in each dish pan
was monitored daily until all larvae/pupae
had died or emerged as adults. Experiments
were repeated three times. If adult mortality
exceeded 30% in control group that test was
either excluded or repeated. Percent emergence inhibition was calculated as described
in Scott et al. (2013).
Mean percent adult emergence inhibition values were pooled for the nine treatment plants and two control plants. Data
were then arcsine square root transformed
to fit the basic assumptions of ANOVA in
JMP version 9 statistical software prior to
analysis. Student’s paired t- tests were used to
determine differences in treatments for each
week compared with controls (P > 0.05).
Adult inhibition in the ASB/pyriproxyfen treatment was about 90% for the first
two weeks of the study. After this time, efficacy gradually decreased to about 80% at week
3 then about 50% during weeks 4 and 5 with
30% adult inhibition at week 6.
Our data suggested that runoff from
pyriproxyfen/ASB treated vegetation can
provide about 90% adult inhibition of Ae.
albopictus for at least 2 weeks after application. The results from this study may serve
as a future model for individuals who want
to combine sugar baits with IGRs for operational larval control in areas where barrier
applications may also be useful for adult
mosquito control.
The authors would like to thank Mike
Smith for technical help. This is a research
report only and the mention of any products
does not imply Anastasia Mosquito Control
District endorsement.
REFERENCES CITED
Ali, A., J. K. Nayar, and R. D. Xue. 1995. Comparative
toxicity of selected larvicides and insect growth regulators to a Florida laboratory population of Aedes
albopictus. J. Am. Mosq. Control Assoc. 11:72-76.
Doud, C. W., A. M. Hanley, K. C. Chalaire, A. G. Richardson, S. C. Britch, and R. D. Xue. 2014. Truckmounted area-wide application of pyriproxyfen targeting Aedes aegypti and Aedes albopictus in northeast
Florida. J. Am. Mosq. Control Assoc. 30:291-297.
Fulcher, A., J. M. Scott, W. A. Qualls, G. C. Muller, and
R. D. Xue. 2014. Attractive toxic sugar baits mixed
Scott et al.: Effects of Leaf Washing On The Persistence of A Sugar Bait-Pyriproxyfen Mixture
with pyriproxyfen sprayed on plants against adult
and larval Aedes albopictus (Diptera: Culicidae). J.
Gerberg et al. 1994. Manual for mosquito rearing and experimental techniques. Bulletin 5. Amer. Mosq. Control
Assoc., Lake Charles, LA pp 102.
Hossain, T. T., A. Fulcher, C. Davidson, J. C. Beier, and
R. D. Xue. 2014. Evaluation of boric acid sugar baits
sprayed on plants against the salt marsh mosquito,
Aedes taeniorhynchus (Diptera: Culicidae). Florida
Entomol. 97:1865-1868.
Muller, G. C., A. Junnila, W. A. Qualls, E. E. Revay, D. L.
Kline, S. A. Allan, Y. Schlein, and R. D. Xue. 2010.
Control of Culex quinquefasciatus in a storm drain
system in Florida using attractive toxic sugar baits.
Naranjo, D. P., W. A. Qualls, G. C. Muller, D. M. Samson,
D. Roque, T. Alimi, K. Arheart, J. C. Beier, and R.
D. Xue. 2013. Evaluation of boric acid sugar baits
against Aedes albopictus (Diptera: Culicidae) in tropical environments. Parasitol. Res. 112:1593-1587.
Qualls, W. A., R. D. Xue, E. E. Revay, S. A. Allan, and
G. C. Muller. 2012. Implications for operational
control of adult mosquito production in cisterns
and wells in St. Augustine, FL using attractive sugar
baits. Acta. Tropica. 124:158-161.
95
Revay, E. E., G. C. Müller, W. A. Qualls, D. L. Kline, D.
P. Naranjo, K. L. Arheart, V. D. Kravchenko, Z. Yefremova, A. Hausmann, J. C. Beier, Y. Schlein, and R.
D. Xue. 2013. Control of Aedes albopictus with attractive toxic sugar baits (ATSB) and potential impact
on non-target organisms in St. Augustine, Florida.
Parasitol. Res. 113:73-79.
SAS Institute, Inc. 2007. JMP. Version 9, Cary, NC.
Scott, J. M., W. A. Qualls, M. K. Gaines, R. D. Xue, C.
Doud, and G. B. White. 2013. Efficacy of ground
ULV application of Nyguard (10% pyriproxifen)
against Aedes albopictus larvae in St Augustine, Florida. Tech. Bull. Florida Mosq. Control Assoc. 9: 4852.
Xue, R. D., G. C. Muller, W. A. Qualls, M. L. Smith, J.
M. Scott, J. Lear, and S. E. Cope. 2013. Attractive
targeted sugar baits: field evaluations and potential
use in mosquito control. Wing Beats. 24:13-18.
Xue, R. D., D. L. Kline, A. Ali, and D. R. Barnard. 2006.
Application of boric acid baits to plant foliage for
22:497-500.
Xue, R. D., G. C. Muller, D. L. Kline, and D. R. Barnard.
2011. Effect of application rate and persistence of boric acid sugar baits applied to plants for control of
Aedes albopictus. J. Am. Mosq. Control Assoc. 27:56-60.
Operational Note
FIELD COMPARISON OF THERMAL FOG APPLICATION OF
DUET® AND BARRIER SPRAYING OF TALSTAR® AGAINST
AEDES ALBOPICTUS AT A RESIDENTIAL PROPERTY IN
ST. AUGUSTINE, FLORIDA
JENNIFER GIBSON, RUI-DE XUE, AND MICHAEL L. SMITH
ABSTRACT. The effectiveness of separate
area-wide ground thermal fog applications
of DUET® (sumithrin/prallethrin) and
barrier spraying of vegetation with Talstar®
(bifenthrin) to control adult Aedes albopictus
populations was compared at a large residential property with dense vegetation and
multiple larval developmental sites in St.
Augustine, Florida. The thermal fog treatment completely eliminated this mosquito
pest through one week post treatment from
a pretreatment landing count that averaged 24/person/3 min. Prior to the barrier
treatment, the average landing rate count
was 28/person/3 min. After application,
landing rate counts were reduced to zero
through three weeks.
imported and 11 locally-acquired cases of
CHIKV. Because Ae. albopictus is quite prevalent in St. Johns County, understanding
and controlling this potential vector species
within local residential areas is becoming increasingly important.
Prior to the 1970’s, area-wide ground
thermal fogging was a mainstay and highly
effective method for adult mosquito control,
especially in the US. Later, this methodology was replaced in favor of ultra low volume technology due to a number of factors
including a rapid spike in the cost of diesel
fuel used as a carrier to deliver the pesticide.
However, compared with ULV application,
thermal foggers showed more consistency
in droplet dispersal across distances as far as
90 m (Britch et al. 2010). Additionally, thermal fog application of pesticides provided
greater mortality against the West Nile virus
vector Culex quinquefasciatus Say (Britch et al.
2010). Recently, Anastasia Mosquito Control
District (AMCD) has re-introduced thermal
fogging into its operational adult control
program (Xue et al. 2013).
During the middle 2000’s, barrier application of residual pesticides to vegetation for
the purpose of controlling adult mosquitoes
gained considerable interest among mosquito control programs, an interest that continues to this day (Trout et al. 2007, Qualls et
al. 2013). By applying residual pesticides to
mosquito harborage/resting sites (e.g. vegetation), mosquito programs can selectively
target adult mosquito species such as Ae. albopictus, a daytime-biting species, at relatively
Key Words. Aedes albopictus, thermal fog, barrier spray, sumithrin, prallethrin, bifenthrin
In 1985, the Asian tiger mosquito, Aedes
albopictus (Skuse) became established in
Houston, Texas after arriving in the USA, as
eggs, in automobile tires imported from Japan (Benedict et al. 2007). Since then, this
mosquito has rapidly spread throughout the
southeastern parts of the USA, including
Florida, and has become the most abundant
and invasive container-inhabiting mosquito
in the State (Lounibos et al. 2001). According to the Centers for Disease Control and
Prevention, Ae. albopictus is a known vector
of multiple arboviruses including chikungunya (CHIKV). In 2014, the Department
of Health confirmed that Florida had 425
96
Gibson et al.: Field Comparison of Thermal Fog Application of Duet and Barrier Spraying of Talstar
low cost (Bengoa et al. 2013). Anastasia Mosquito Control District has used insecticide
barrier applications to perimeter vegetation
to successfully control adult mosquitoes
around golf courses and schools (Qualls et
al. 2013).
The objective of our study was to compare the effectiveness of an area-wide thermal fog application of DUET® with that of
a barrier application of Talstar®, against the
container-inhabiting mosquito, Ae. albopictus, at a large local residential property.
Pre-treatment landing rate counts (LRC)
were conducted by three volunteers for
three minutes in three different locations
approximately 35 meters apart at a 2.7 hectare County St. Johns County property (latitude 29.93049, longitude -81.343876). The
property was chosen due to its numerous larval developmental sites and dense vegetation
that are known habitat locations of adult Ae.
albopictus. The study took place from 0900 to
1100 during October 2014 when peak seasonal populations of this species occur in St.
Johns County.
DUET (active ingredient: 5% sumithrin® and 1% prallethrin) was applied
at maximum label rate along the outside
perimeter of the property and domicile using a LongRay® TS-35A thermal fogger
(ADAPCO, Sanford, FL). The travel distance
at which treatment was applied at was 11 m
per minute at a spray height of 3 meters.
Wind direction and speed were monitored
during application. At 30 minutes, and 24
hours after treatment, landing rate counts
were conducted again for three minutes in
the same locations used for pre-treatment.
Landing rate counts were conducted weekly
thereafter and continued until there was a
significant increase in post treatment Ae. albopictus LRC.
Once Ae. albopictus LRCs rebounded
after two weeks, a residual barrier application was administered, at maximum label
rate, to perimeter vegetation of the property
with Talstar® (active ingredient: 7.9% bifenthrin) using a Birchmeier Rec 15 backpack
sprayer. The sprayer was calibrated to deliver
43.3 ounces per minute with a mixture of 3
gallons of water to 3 ounces of Talstar. The
97
application rate was 30 meters per 3 minutes
at a spray height of 3 meters. Landing rate
counts were conducted 24 hours, 1 week, 2
weeks, and 3 weeks after treatment.
Mean landing rate counts were subjected
to analysis using JMP® software (SAS Institute, Inc. 2007). Student’s t-test was used
to compare pre- and post treatment LRC’s
within each treatment (i.e. thermal fog and
barrier).
Our results showed that there was a significant difference between LRCs pre and
post for the thermal fog treatment after 24
hours (t = 2.15, P < 0.05). This application
completely eliminated the Ae. albopictus population through 1 week post treatreatment
from a pretreatment LRC that averaged 24/
person/3 min. For the barrier application,
pre- and post LRCs were also significantly
different after 24 hours (t = 2.15, P < 0.05).
The pre-treatment LRC averaged 28 Ae. albopictus/person/3 min, then after application, LRCs were reduced to zero through 3
weeks post treatment.
Targeting an urban mosquito vector for
control, like Ae. albopictus, can be challenging due to high population densities of the
pest often produced from numerous cryptic
larval habitats. Determining adult harborage sites to target control measures can be
helpful in designing the scenario for barrier
application. For example, one study showed
that Ae. albopictus had high mortality in leaf
bioassays after exposure to Talstar-treated
wax myrtle plants (Cilek and Hallmon 2008).
Plant identification can provide an indicator for harborage sites and is only one of
the many variables that integrated mosquito management programs need to address
when considering barrier application effectiveness. Past studies by Cilek (2008) have
shown that some pyrethroid insecticides,
applied as barrier treatments to vegetation,
can last up to 6 weeks in controlling adult
mosquitoes. Our results suggest that the barrier treatment was a better alternative for
controlling Ae.albopictus in dense vegetation
because of its longer lasting effects on the
population in the field. However, we believe
that by utilizing a combination of methods,
such as thermal fogging and barrier spray-
98
ing, mosquito control districts may be able
to limit costs in areas where consistent adult
control is needed but may be difficult to
achieve. Furthermore, AMCD will continue
to judiously use thermal fogging and barrier
treatments as only one of the tools in its integrated mosquito management program in
order to prevent the outbreak of mosquitoborne diseases in St. Johns County.
Special thanks go to the owners of the
residence that allowed us conduct this study
on their property. We are also thankful for
the support of Ali Fulcher, Cat Smith and
the AMCD volunteers who helped conduct
the landing rate counts.
REFERENCES CITED
Benedict, M. Q., R. S. Levine, W. A. Hawley, and L. P.
Lounibos. 2007. Spread of the tiger: global risk of invasion by the mosquito Aedes albopictus. Vector-borne
Zoonotic. Dis. 7:76-85.
Bengoa, M., R. Eritja., and J. Lucientes. 2013. Laboratory tests of the residual effect of deltamethrin on
vegetation against Aedes albopictus. J. Am. Mosq. Control Assoc. 29:284-288.
Britch, S. C., K. J. Linthicum, W. W. Wynn, T. W. Walker, M. F. Farooq, V. L. Smith, C. A. Robinson, B. B.
Lothrop, M. Snelling, A. Gutierrez, H. D. Lothrop,
J. D. Kerce, J. J. Becnel, U. R. Bernier, and J. W. Pridgeon. 2010. Evaluation of ULV and thermal fog mosquito control applications in temperate and desert
environments. J. Am. Mosq. Control Assoc. 26:183197.
Cilek, J. E. 2008. Application of insecticides to vegetation as barriers against host-seeking mosquitoes. J.
Cilek, J. E. and C. F. Hallmon. 2008. Residual effectiveness of three pyrethroids on vegetation against
adult Aedes albopictus and Culex quinquefasciatus in
screened field cages. J. Am. Mosq. Control Assoc.
24:263-269.
Lounibos, L. P., G. F. O’Meara, R. L. Escher, N. Nishimura, M. Cutwa, T. Nelson, and S. A. Juliano. 2001.
Testing predictions of displacement of native Aedes
by the invasive Asian tiger mosquito Aedes albopictus
in Florida, USA. Biol. Invasions 3:151-166.
Qualls, W. A., M. L. Smith, and R. D. Xue. 2013. Successful applications of barrier treatments using bifenthrin against mosquitoes in St. Johns County,
Florida, from 2006-2009. Tech. Bull. Florida Mosq.
SAS Institute, Inc. 2007. JMP®. version 11. Cary, NC.
Trout, R. T., G. C. Brown, M. F. Potter, and J. L. Hubbard. 2007. Efficacy of two pyrethroid insecticides
applied as barrier treatments for managing mosquito (Diptera: Culicidae) populations in suburban residential properties. J. Med. Entomol.
44:470-477.
Xue, R. D., Weaver, J. H., M. L. Smith, and W. A. Qualls.
2013. Field evaluation of thermal fog application
of sumithrin products against adult mosquitoes in
northeast Florida. Tech. Bull. Florida Mosq. Control
Assoc. 9:38-41.
The Eleventh Arbovirus Surveillance and
Mosquito Control Workshop
Sponsored by AMCD and USDA/CMAVE
March 25-27, 2014
PROGRAM
Tuesday (March 25, 2014)
Panel Session:
Moderator: Dr. Rui-De Xue, Director, AMCD, St. Augustine, FL
8:30 am
Welcome & Introduction
Mrs. Catherine Brandhorst, Chairperson, AMCD Board of Commissioners
Dr. Ken Linthicum, Center Director, USDA/CMAVE
Mr. Neil Wilkinson, President, Florida Mosquito Control Association
8:40 am
Keynote Speaker: New insights into the ecology of eastern equine encephalitis virus transmission in the southeastern USA ...
Dr. Thomas Unnasch, Distinguished USF Health Professor and Department Chair, University of South Florida, Tampa, FL
9:20 am
Dengue vector control in Malaysia
Dr. Abu Ahmad, Professor, Universiti Sains, Malaysia
9:45 am
The transgenic mosquitoes and its release for dengue control in Brazil
Dr. Aldo Malavasi, Director, Medfly and Mosquito Facility, Brazil
10:10 am
Overview of dengue vaccine study in China
Dr. Qiang-Ming Sun, Visiting Professor, Yale University, New Haven, CT
10:35 am
Mosquitoes and plants—the principle and use of plants in mosquito
control
Dr. Rui-De Xue, Director, AMCD, St. Augustine, FL
10:45 am
Break
Moderator: Dr. Ken Linthicum, Center Director, USDA/CMAVE, Gainesville, FL
11:00 am
10:20 am
11:40 am
Real time monitoring of the dengue vector online: cost benefit of a Brazilian technology
Dr. Alvaro E. Eiras, Professor, Universidade Federal de Minas Gerais,
Belo Horizonte, Brazil
Historical perspective of dengue fever and its management control strategy in Saudi Arabia
Dr. Abdul Aziz Althbyani, Assistant Professor, University of Tabuk, Saudi
Arabia
Challenges for the malaria vector control strategies in three ecological
zones (West, Central and East Africa)
99
100
Dr. Seydou Doumbia, Professor and Co-Director of MRTC/FMPOS,
University of Sciences, Techniques and Technology of Bamako, Mali
12:00 pm
An update about Clarke’s new products for mosquito control
Mr. Frank Clarke, Vice President, Clarke, Kissimmee, FL
12:10 pm
Lunch break (provided by Clarke)
Programs and Associations:
Moderator: Dr. Gary Clark, Research Leader, USDA/CMAVE, Gainesville, FL
1:00 pm
The International Center of Excellence in Research of Vector-borne
Diseases in Mali
Dr. Traore Sekou F., Professor and Co-Director of MRTC/FMPOS, University of Sciences, Techniques and Technology of Bamako, Mali
1:20 pm
Overview of USDA/CMAVE program
Dr. Ken Linthicum, Center Director, USDA/CMAVE, Gainesville, FL
1:40 pm
USDA/Rutgers University’s area wide management program of (http://
asiantigermosquito.rutgers.edu/)
Dr. Gary Clark, Research Leader, USDA/CMAVE, Gainesville, FL
2:00 pm
Update on the American Mosquito Control Association
Dr. Roxanne Connelly, President, AMCA, Professor, University of
Florida/IFAS/FMEL, Vero Beach, FL
2:15 pm
Overview of the Society of Vector Ecology
Dr. Dan. Kline, Vice President, SOVE, Research Entomologist, USDA/
ARS/CMAVE, Gainesville, FL
2:30 pm
Update on the Florida Mosquito Control Association
Mr. Neil Wilkinson, President, FMCA, Florida Gulf Coast University, Ft.
Myers, FL
2:45 pm
Chemical and biological threats and vulnerabilities in industry and academia
Mrs. Bridgette Frost, FBI Special Agent, Jacksonville, FL
3:05 pm. Break
Arbovirus/Dengue Outbreak in Martin County, FL:
Moderator: Dr. John Beier, Professor, University of Miami, Miami, FL
3:20 pm
Arbovirus surveillance in Florida, 2013
Stephanie M. Moody-Geissler, Arbovirus Surveillance Coordinator, Bureau of Epidemiology, FLDOH, Tallahassee, FL
3:40 pm
Dengue outbreak in Martin County, Florida in 2013
Mrs. Karlette Peck, Martin County Department of Health, Stuart, FL
4:00 pm
FDACS’s entomological investigation on the 2013 outbreak of dengue
fever in Martin County
Dr. Peter Jiang, Medical Entomologist, Bureau of Pesticides, FDACS,
Tallahassee, FL
4:15 pm
Larval surveillance of dengue fever vectors, and in Hillsborough
County, FL, 2011-2012
Dr. Azliyati Azizan, Assistant Professor, Department of Global Health,
University of South Florida, Tampa, FL
Program101
4:35 pm
Analysis of sentinel chickens and arbovirus in St. Johns County, FL
Mr. Richard Weaver, Data Manager, AMCD, St. Augustine, FL
4:45 pm
WNV in Jacksonville, Florida, 2013 and activity pattern of vector
mosquitoes
Ms. Marah Clark, Entomologist, Jacksonville Mosquito Control, FL
5:00 pm
Cooperative projects to control in Florida and Thailand
Drs. Kenneth Linthicum and Seth C. Britch, USDA/CMAVE, Gainesville,
FL
5:20 pm
End of the session
6:00 pm
Dinner and Lecture (Holiday Isle, Beach Blvd, St. Augustine Beach
City): Highlights of the Oldest City, City Commissioner, St. Augustine,
FL
Wednesday (March 26, 2014)
Moderator: Dr. Dan Kline, Research Entomologist, USDA/CMAVE, Gainesville, FL
8:30 am
Sex, Y, and the control of mosquito-borne infectious diseases
Dr. Jake Tu, Professor, Virginia Tech University, Blacksburg, VA
8:50 am
Gene silencing as a strategy for vector control
Dr. James Becnel and Mr. Al Estep, USDA/CMAVE, Gainesville, FL
9:10 am
Update about research on carbonic anhydrases in mosquitoes
Mr. D. Dixon and Dr. Paul Linser, Professor, Whitney Laboratory, University of Florida, St. Augustine, FL
9:30 am
Integrating molecular tools with Florida mosquito control
Dr. Liming Zhao, Research Assistant Professor, University of Florida/
IFAS/FMEL, Vero Beach, FL
9:50 am
Environmental assessment of mosquito ecology in Northern Haiti
Dr. Whitney A. Qualls, Ms D.M. Samson, and Dr. John Beier, University
of Miami, FL
Expanding integrated vector management to promote healthy environments
Dr. John Beier, Professor, University of Miami, Miami, FL
10:30 am
Break
10:10 am
Attractants/Traps/Repellents:
Moderator: Dr. Uli. Bernier, Research Chemist, USDA/CMAVE, Gainesville, FL
10:40 am
Developing new insecticides and repellents for skin and cloth
Dr. Uli. Bernier, Research Chemist, USDA/CMAVE, Gainesville, FL
11:00 am
Evaluation of spatial repellents for military use
LT Marcus McDonough, Navy Entomology Center of Excellence, Jacksonville, FL
11:15 am
Plant flower attraction for Aedes albopictus
Dr. Dan. Kline, Research Entomologist, USDA/CMAVE, Gainesville, FL
11:35 am
Field assessment of yeast and oxalic acid generated CO2 for mosquito
surveillance
LT James Harwood, Navy Entomology Center of Excellence, Jacksonville, FL
102
11:50 am
Five light trap comparison for collection of mosquitoes and non-target
insects
Dr. Chun-Xiao Li, Visiting Scientist, Department of Entomology and
Nematology, University of Florida, Gainesville, FL
12:00 pm
An update about ADAPCO’s new technology
Mr. Derek Wright, ADAPCO, Sanford, FL
12:15 pm
Lunch Break (provided by ADAPCO)
Larval Habitats & Control:
Moderator: Dr. Jerry Hogsette, Research Entomologist, USDA/ARS/CMAVE, Gainesville,
FL
1:00 pm 1:20 pm
Florida’s Indian River Lagoon: mosquito control within an estuary in
distress
Mr. Doug Carlson, Director, Indian River Mosquito Control District,
Vero Beach, FL
Vertical distribution of container-inhabiting mosquitoes in a LaCrosse
virus endemic area
Mr. Michael Riles, Dr. Bruce Harrison, and Dr. Brian Byrd, University of
Western North Carolina, NC
1:30 pm
Acoustic techniques for mosquito control in Houston, TX
Mr. Herbert Nyberg, New Mountain Innovations, Inc., Niantic, CT
1:50 pm
The use of cree carrying larvicides for possible control of mosquito
larvae
Mr. John Olsen, President, Cree Industries Inc., FL
2:10 pm
Aerial larviciding in the Florida Keys
Mr. Mike Doyle, Director, Florida Keys Mosquito Control District, Key
West, FL
2:30 pm
Evaluation of Mosquiron CRD against in downtown storm drains, St.
Augustine
Mrs. Ali Fulcher, Biologist, AMCD, St. Augustine, FL
2:40 pm
A durable dual action lethal ovitrap for control of container breeding
mosquitoes
Drs. Roberto Pereira and Phil Koehler, Department of Entomology and
3:00 pm
Wildlife lighting- protect wildlife through responsible lighting practices
Mr. Mike Hudon, Research Entomologist, Indian River Mosquito Control District, Vero Beach, FL
3:20 pm
Break
Adult Control:
Moderator: Dr. Jake Tu, Professor, Virginia Tech University, Blacksburg, VA
Primarily testing and formulations of an attractive targeted sugar bait
against Aedes aegypti and Aedes albopictus
Ms. Jodi Scott, Education Specialist, AMCD, St. Augustine, FL
3:50 pm
Hand thermal fogging comparison study
Mrs. Ali Fulcher, Biologist, AMCD, St. Augustine, FL
3:40 pm
Program103
4:00 pm
Sticky traps against Aedes aegypti
Ms. Emily Thompson AMCD, St. Augustine, FL and Ms. Jodi Scott, Department of Entomology and Nematology, University of Florida, Gainesville, FL
4:10 pm
Exploring new thermal fog and ultra-low volume technologies to improve indoor control of the dengue vector,
LT James Harwood, Navy Entomological Center of Excellence,
Jacksonville, FL
4:25 pm
The knight stick: a new trap for stable flies
Dr. Jerry Hogsette, Research Entomologist, USDA/ARS/CMAVE,
Gainesville, FL
4:45 pm
Attractive targeted sugar baits (ATSB)
Mrs. Julie Lear, Marketing Director, Universal Pest Solutions, Dallas, TX
4:55 pm
Attractive toxic sugar bait stations for malaria vector control in Mali
Mr. Mohamed M. Traore, University of South Carolina, Colombia, SC
5:10 pm
Aerial spray to control Aedes aegypti in Manatee County, FL
Mr. Mark Latham, Director, Manatee County Mosquito Control District, FL
5:30 pm
End of the session
Thursday (March 27, 2014)
Programs, Education, & Legislation:
Moderator: Mr. Neil Wilkinson, Florida Gulf Coast University, Ft. Myers, FL
8:30 am
Overview of the school program at Lee County Mosquito Control District
Mr. Neil Wilkinson, Florida Gulf Coast University, Ft. Myers, FL
8:50 am
Efficacy of a new fly bait containing cyantraniliprole
Mr. Casey Parker, Drs. Roberto Pereira and Phil Koehler, Department
of Entomology and Nematology, University of Florida, Gainesville, FL
9:00 am
ATSB and pyriproxfen on plants against adult and larval Aedes aegypti
9:15 am
FLDACS update – progress on rulemaking
Mr. Michael Page, Chief, Bureau of Entomology and Pest Control, FLDACS, Tallahassee, FL
9:30 am
Overview of Mosquito Research Foundation
Mr. James Clauson, Director, Beach Mosquito Control District, Panama
City, FL
9:45 am
Program overview of Volusia County Mosquito Control
Mr. James McNelly, Director, Volusia Mosquito Control District, New
Smyrna Beach, FL
10:00 am
South Walton County Mosquito Control program
Mr. Ben Brewer, Director, South Walton County Mosquito Control District, Santa Rosa Beach FL
10:20 am
Break
104
Surveillance, Control, & Other:
Moderator: Mr. Michael Page, Chief, Bureau of Entomology and Pest Control, FLDACS,
Tallahassee, FL
10:35 am
10:45 am
Large scale evaluation of a new barrier spray machine and barrier treatment
Mr. Mike Smith, Biological Technician, AMCD, St. Augustine, FL
Early population outbreak of mosquitoes and control response in St.
Johns County, FL
Mrs. Kay Gaines, Supervisor, AMCD, St. Augustine, FL
10:55 am
Evaluating the temporal effects of Nuvan Prostrips for filth fly control
LCDR Shani Gourdine, Navy Entomological Center for Excellence,
Jacksonville, FL
11:10 am
Evaluation of DeltaGard G as an acaricide
LT Mattew Yans, Navy Entomological Center for Excellence, Jacksonville, FL
11:25 am
Database software with smart phone for operational mosquito control
Mr. Tim Morris, Mobisoft Infotech, Houston, TX
11:40 am
Spot weather detectors & recorder and possible for mosquito control
operation
Dr. William R. Eisenstadt, Professor, Department of Electrical and Computer Engineering, University of Florida, Gainesville, FL
12:00 pm
Update about products from UNIVAR
Mr. Jason E. Conrad, Industry Specialist, UNIVAR
12:10 pm
Lunch break (provided by UNIVAR)
END OF WORKSHOP
The Twelfth Arbovirus Surveillance and
Mosquito Control Workshop
Sponsored by AMCD and USDA/CMAVE
March 24-26, 2015
PROGRAM
Tuesday, March 24, 2015
Panel Session:
Moderator:Dr. Rui-De Xue, Director, AMCD, St. Augustine, FL
8:30 am
Welcome & Introduction
Ms. Vivian Browning, Chairperson, AMCD Board of Commissioners
Dr. Ken Linthicum, Center Director, USDA/CMAVE, Gainesville, FL
Ms. Sue Bartlett, President of the FMCA, New Smyrna, FL
8:40 am
Keynote Speaker: Epidemic of chikungunya and dengue fever and research response and focus from the University of Miami
Dr. John C. Beier, Professor and Section Chief, University of Miami,
Miller School of Medicine, Miami, FL
9:20 am Guest Speaker: Overview of Rutgers University’s Center for Vector Biology
Dr. Randy Gaugler, Distinguished Professor and Director, Center for
Vector Biology, Rutgers University, NJ
9:50 am
Dengue virus and potential vaccine investigation
Dr. Scott Michael, Professor, Florida Gulf Coast University, Ft. Myers, FL
10:20 am
The 4th International Forum for Surveillance and Control of Mosquitoes
and Mosquito-borne Diseases, Guangzhou, China, (May 25-28, 2015)
and AMCD’s new complex
10:30 am
Break
Moderator: Dr. Ken Linthicum, Center Director, USDA/CMAVE, Gainesville, FL
11:00 am
11:20 am
Spatial Repellents for Control of Vector-borne Disease: The Current
Status
Dr. Nicole Achee, Associate Professor, University of Notre Dame, IN
Progress and prospective of attractive toxic sugar baits against mosquitoes and sand Flies
Dr. Gunter Muller, Visiting Professor, Hebrew University, Jerusalem,
Israel
Extreme weather events and impact on vector-borne diseases and agriculture
Dr. Kenneth Linthicum, Center Director, USDA/CMAVE, Gainesville, FL
12:00 pm
An update on Clarke’s new products for mosquito control
Mr. Frank Clarke, Vice President, Clarke, Kissimmee, FL
12:10 pm
Lunch break (provided by Clarke)
11:40 am
105
106
Programs & Associations:
Moderator: Dr. Dan Kline, Research Entomologist, USDA/CMAVE, Gainesville, FL
1:00 pm
ATSB in the marketplace, commercial and consumer update
Ms. Laura Uggerholt, Universal Pest Control
1:20 pm
FLDACS program update
Ms. Adriane N. Tambasco, Medical Entomologist, FLDACS/Division of
Agricultural Environmental Services, Tallahassee, FL
1:40 pm
Update on the American Mosquito Control Association in 2015
Dr. Ken Linthicum, President, AMCA and Center Director, USDA/
CMAVE, Gainesville, FL
1:55 pm
Overview of the Society of Vector Ecology
Dr. Dan Kline, President-Elect, SOVE and Research Entomologist,
USDA/ARS/CMAVE, Gainesville, FL
2:10 pm
Update on the Florida Mosquito Control Association
Mrs. Sue Bartlett, President, FMCA and Operations Manager, Volusia
Mosquito Control, New Smyrna, FL
2:25 pm
Overview of the Florida Entomological Society
Dr. Cindy McKenzie, President, Florida Entomological Society and Research Entomologist, USDA/USHRL, Ft. Pierce, FL
2:40 pm
Malaria vector mosquito’s resting behavior and movement in day time
Dr. Gunter Muller, Visiting Professor, Hebrew University, Jerusalem,
Israel
3:00 pm
Break
Arbovirus:
Moderator: Dr. Randy Gaugler, Distinguished Professor and Director, Rutgers University, NJ
3:20 pm
Arbovirus surveillance in Florida, 2014: the year of chikungunya
Dr. Andrea Bingham, Arbovirus Surveillance Coordinator, Bureau of
Epidemiology, FLDOH, Tallahassee, FL
3:40 pm
Long-term health impacts of chikungunya virus infection in Florida
residents
Ms. Katie Kendrick, Epidemiologist, Bureau of Epidemiology, FLDOH,
Tallahassee, FL
4:00 pm
West Nile virus outbreak in Volusia County, Florida in 2014
Mr. James McNelly, Director, Volusia Mosquito Control, New Smyrna,
FL
4:20 pm
Vector competence of Florida mosquitoes for chikungunya and dengue
viruses
Dr. Barry Alto, Assistant Professor, University of Florida/IFAS, Florida
Medical Entomology Laboratory, Vero Beach, FL
4:40 pm
Arbovirus surveillance by sentinel chickens and a novel technique in St.
Johns County
Miss Jennifer Gibson, Biological Technician, AMCD, St. Augustine, FL
West Nile virus in Jacksonville, Florida, 2014 and daily activity pattern of
vector mosquitoes
Ms. Marah Clark, Entomologist, Jacksonville Mosquito Control District, FL
5:00 pm
Program107
5:20 pm
End of the session
6:00 pm
Dinner and Lecture (Holiday Isle, Beach Blvd., City of St. Augustine
Beach): Multi-rotor unmanned aerial systems: new tool for mosquito
control
Dr. Randy Gaugler, Distinguished Professor and Director, Rutgers University’s Center for Vector Biology, NJ
Wednesday, March 25, 2015
Biology & Ecology:
Moderator: Dr. Nicole Achee, Associate Professor, University of Notre Dame
8:20 am
Update on the Acoustic Techniques for Mosquito Control
Mr. Herbert Nyberg, New Mountain Innovations, Inc., Niantic, CT
8:35 am
Mosquito gut ecosystem and potential impact on control action
Dr. Jiannong Xu, Associate Professor, Department of Biology, New
Mexico State University, Las Cruces, NM
8:55 am
Population dynamics of the floodwater mosquito in St. Johns County,
Florida
Miss Emily Thomson, Biological Technician, AMCD, St. Augustine, FL
9:05 am Aedes japonicus japonicus (Theobald): a new emerging vector in northwest Florida
Mr. Mike Riles, Entomologist, Beach Mosquito Control District,
Panama City Beach, FL
9:20 am
Update on research on carbonic anhydrases in Aedes aegypti
Mr. D. Dixon and Dr. Paul Linser, Whitney Laboratory, University of
Florida, St. Augustine, FL
9:35 am
Mosquitoes and non-target insects and attractive target sugar baits
Ms. Jodi Scott, Visiting Scientist, AMCD, St. Augustine, FL and Ph.D.
candidate, Department of Entomology and Nematology, University of
Florida, Gainesville, FL
Mosquitoes associated with suburban backyards and dog kennels near
Gainesville, FL
Mr. Chris Holderman, Ph.D. candidate, Department of Entomology and
9:50 am
10:05 am
Comparison of target and non-target mortality rates from residual pesticide on HESCO material
Mr. Robert L. Aldridge, Mr. Thomas T. Dao, Dr. Seth C. Britch and Dr.
Kenneth J. Linthicum, USDA/CMAVE, Gainesville, FL
10:20 am
Indoor and outdoor sugar feeding behaviors of dengue vector in Ecuador
Drs. Whitney Qualls and John Beier, Professor, University of Miami,
Miami, FL
10:35 am
Break
Attractants/Traps/Repellents:
Moderator: Dr. James Becnel, Research Entomologist, USDA/CMAVE, Gainesville, FL
10:40 am
Current research on mosquito repellents for clothing and skin
Ms. Natasha Agramonte and Dr. Ulrich Bernier, USDA/CMAVE, Gainesville, FL
108
11:00 am
Various approaches for the control of blood-feeding insects
Dr. Lisa L. Drake, Department of Biology, New Mexico State University,
Las Cruces, NM
11:20 am
A multiyear study of oviposition patterns in suburban residential backyards
Dr. Dan Kline, Research Entomologist, USDA/CMAVE, Gainesville, FL
Evaluation of thermal cell mosquito repellents against several species of
mosquitoes
Mr. Christopher Bibbs, Education Specialist, AMCD, St. Augustine, FL
12:00 pm
An update on ADAPCO’s new products
Mr. Derek Wright, ADAPCO, Sanford, FL
12:10 pm
Lunch break (provided by ADAPCO)
11:40 am
Pesticides, Larval &Adult Control:
Moderator: Dr. Jerry Hogsette, Research Entomologist, USDA/ARS/CMAVE, Gainesville, FL
1:00 pm
Ground application of larviciding and adulticiding at AMCD
Mrs. Kay Gaines, Supervisor, AMCD, St. Augustine, FL
1:15 pm
New formulations of larvicide and adulticide for mosquito control ….
Mr. David Sykes, All Pro Vector Group, Madison, FL
1:25 pm
Effects of residual IGRs on Aedes aegypti and Aedes albopictus
Ms. Kristen Donovan, Undergraduate Research Assistant, Dr. Roberto
Pereira and Dr. Phil Koehler, Associate Research Scientist and Professor,
Entomology Department, University of Florida, Gainesville, FL
1:40 pm
Malaria transmission along the Niger river in a Sudan savanna area of Mali
Dr. Sekou F. Trtaore, Professor and Director of MRTC Entomology,
University of Sciences, Techniques and Technology of Bamako, Mali
1:50 pm
Program Overview: Collier Mosquito Control District
Mr. Jim Stark, Executive Director, Collier Mosquito Control District,
Naples, FL
2:10 pm
Evaluation of larvicides and pupacides against Aedes aegypti and Culex
quinquefasciatus
Miss Jennifer Gibson, Biological Technician, AMCD, St. Augustine, FL
2:20 pm
Public Health Pesticide Inventory and new insecticides
Dr. Karl Malamud-Roam, Public Health Pesticides Program Manager,
IR-4 Project Headquarters, Princeton, NJ
2:40 pm
Possible impact of mosquito control pesticides on coral reef health in
Florida
Dr. Cliff Ross, Associate Professor, Department of Biology, University of
North Florida, Jacksonville, FL
3:00 pm
3:20 pm
Evaluation of Cimi-Shield Knock-Out Bed Bug Eliminator against adult
house flies
Dr. Jerry Hogsette, Lead Scientist and Research Entomologist, USDA/
ARS/CMAVE, Gainesville, FL
Break
Program109
Insecticide Resistance:
Moderator: Dr. Whitney Qualls, University of Miami, Miami, FL
3:40 pm
Resistance mechanisms in the Akron strain of Anopheles gambiae isolated in
Benin, West Africa
Drs. J. R. Bloomquist, A. D. Gross, D. R. Swale, F. Tong, and T. D. Anderson, Department of Entomology and Nematology, Emerging Pathogens Institute, University of Florida, Gainesville, FL
4:00 pm
Characterization of a pyrethroid resistant atrain of from Puerto Rico
Dr. James J. Becnel, Mr. Alden Estep, Mr. Neil Sanscrainte and Ms. Jessica Louton, USDA/CMAVE, Gainesville, FL
4:20 pm
Survey of Aedes aegypti in Florida to determine resistance levels and
mechanisms
Mr. Alden Estep, Dr. James J. Becnel, and Mr. Neil Sanscrainte, USDA/
CMAVE, Gainesville, FL and Ms Christine Waits, Navy Entomology Center of Excellence
4:40 pm
Comparison of pyrethroid resistance in adult and larval Culex pipiens
pallens in five field populations from China
Ms. Hui Liu, Research Associate, Department of Vector Biology and
Control, Jiangsu Provincial CDC, Nanjing, China
Testing for insecticide resistance in Aedes alboptictus strains from St.
Augustine, Florida
Ms. Christy Waits, Navy Entomology Center for Excellence, Jacksonville,
FL, Mrs. Ali Fulcher, AMCD, Mr. Alden Estep, Mr. Alec Richardson,
Navy Entomology Center for Excellence, Jacksonville, FL, Dr. Jimmy
Becnel, USDA/CMAVE, and Dr. Rui-De Xue, AMCD
5:00 pm
5:20 pm
Vector epidemiology and challenge of malaria control in Mali
Dr. Seydou Doumbia, Professor and Director, MRTC Entomology, University of Sciences, Techniques and Technology of Bamako, Mali
5:40 pm
End of the session
Thursday, March 26, 2015
Technology & Program:
Moderator: Ms. Sue Bartlett, Operations Manager, Volusia Mosquito Control, New Smyrna, FL
8:30 am
SMACK those mosquitoes: a “Sentinel Mosquito Arbovirus Capture Kit”
(SMACK) for monitoring arboviruses in the field
Dr. Scott Ritchie, Professor, James Cook University, Cairns, Australia
9:00 am
Overwintering of eastern equine encephalitis virus in North America
Dr. Nathan Burkett-Cadena, Assistant Professor, University of Florida/
9:20 am
Durable Dual-action Lethal Ovitraps (DDALO) for management of the
dengue vector and other container-breeding mosquitoes
Ms. Casey Parker, Graduate Research Assistant, Drs. Roberto Pereira and
Phil Koehler, Associate Research Scientist and Professor, Department of
Entomology and Nematology, University of Florida, Gainesville, FL
9:35 am
Overview of the IGR-pyriproxyfen through auto-dissemination against
dengue vector mosquitoes
Mr. Mike Banfield, CEO, Spring Star, Inc. Seattle, WA
110
9:50 am
Database software with smart phone for operational mosquito control in
AMCD
Mr. Richard Weaver, Data Manager, AMCD and Mr. Tim Morris, Mobisoft Infotech, Houston, TX
10:05 am
GeoWorld Outdoors, Inc. GeoMosquito Mapping Solution
Mr. Daniel Finch, GeoWorld Outdoors, Inc, Kenosha, WI
10:20 am
Mobile application with a focus on pest activity, material target pests,
and e-content
Mr. Adam Holt, Service Pro, St. Augustine, FL
10:30 am
Break
Surveillance & Other:
Moderator: Dr. Nathan Burkett-Cadena, Assistant Professor, UF/IFAS/FMEL, Vero Beach
10:50 am
11:00 am
Population surveillance of Aedes albopictus detected by BGS traps in St.
Johns County, FL
Mr. Mike Smith, Biologist, AMCD, St. Augustine, FL
Development of a rapid detection method for arbovirus surveillance in
Florida
Dr. Limin Zhao, Research Assistant Professor, University of Florida/
11:20 am
Understanding IGRs and their potential for controlling bed bugs
Ms. Brittany Delong, Graduate Research Assistant, Drs. Roberto Pereira
and Phil Koehler, Associate Research Scientist and Professor, Department of Entomology and Nematology, University of Florida, Gainesville,
FL
11:40 am
Mosquito population surveillance and GIS
Dr. Mohamed F. Sallam, Visiting Scientist, Department of Entomology
and Nematology, University of Florida, Gainesville, FL
12:00 pm
Intact Immunoglobulin traffic from blood meal to hemoplymph: a new
look at mosquito control via host immunization
Dr. Paul Linser, Professor, Whitney Laboratory, University of Florida, St.
Augustine, FL
12:20 pm
Update on products from UNIVAR
Mr. Jason E. Conrad, Industry Specialist, UNIVAR
12:30 pm Lunch break (provided by UNIVAR)
END OF WORKSHOP
2:30 pm
New AMCD facility ground breaking

TECHNICAL BULLETIN OF THE FLORIDA MOSQUITO CONTROL

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