Hematology, blood chemistry and plasma

Transcription

Veterinary Clinical Pathology ISSN 0275-6382
ORIGINAL RESEARCH
Hematology, leukocyte cytochemical analysis, plasma
biochemistry, and plasma electrophoresis of wild-caught and
captive-bred Gila monsters (Heloderma suspectum )
Kerri Cooper-Bailey1, Stephen A. Smith1, Kurt Zimmerman1, Rosalie Lane2, Rose E. Raskin3, Dale DeNardo4
1
Department of Biomedical Sciences and Pathobiology, Virginia–Maryland Regional College of Veterinary Medicine, Virginia Polytechnic Institute and
State University, Blacksburg, VA, USA; 2Avian and Exotic Animal Clinical Pathology Laboratory, Wilmington, OH, USA; 3Department of Comparative
Pathobiology, School of Veterinary Medicine, Purdue University, West Lafayette, IN, USA; and 4School of Life Sciences, Arizona State University, Tempe,
AZ, USA
Key Words
Blood values, CBC, cytochemistry,
hemoparasites, lizard
Correspondence
Stephen A. Smith, Department of Biomedical
Sciences and Pathobiology, Virginia–Maryland
Regional College of Veterinary Medicine,
Virginia Polytechnic Institute and State
University, Duck Pond Drive, Phase II,
Blacksburg, VA 24061-0442, USA
E-mail: [email protected]
DOI:10.1111/j.1939-165X.2011.00337.x
Background: The Gila monster (Heloderma suspectum), one of several venomous lizard species in the world, is found within the United States and
Mexico and is recognized as an iconic symbol of the American Southwest.
Thus, Gila monsters are of growing interest in the captive reptile trade and
within zoological and educational institutions.
Objectives: The aims of this study were to determine results for CBCs,
describe cytochemical reactions in WBCs, and obtain plasma biochemical and
protein electrophoresis results from wild-caught and captive-bred H. suspectum.
Methods: Ventral tail vein blood samples were obtained from 16 captive
(14 wild-caught and 2 captive-bred) Gila monsters. CBCs, RBC morphometric analysis, plasma biochemical analysis, and protein electrophoresis
were performed. Leukocytes were stained for peroxidase, Sudan black B
(SBB), chloroacetate esterase, napthyl butyrate esterase, and leukocyte
alkaline phosphatase, and blood smears were examined for the presence of
hemoparasites.
Results: The median (range) PCV was 37% (22–50%) and WBC count was
4.6 103/mL (3.3–6.4 103/mL) with approximately 50% heterophils and
fewer lymphocytes, basophils, azurophils, and monocytes in decreasing
order. Cytochemical reactions were unique among reptiles with strong
staining for peroxidase and SBB in monocytes/azurophils. Biochemical
results were similar to those of earlier reports with slight increases in uric
acid and urea concentrations. Plasma electrophoretic results indicated that
albumin was approximately equal to the combined globulin fractions.
Conclusions: Results of blood analysis in healthy wild-caught and captivebred H. suspectum may be useful for monitoring health status in this species.
Introduction
Materials and Methods
The Gila monster, Heloderma suspectum, is one of several
species of venomous lizards in the world.1 Its distribution is tightly linked to the Sonoran Desert located
within Arizona and the state of Sonora, Mexico, and its
range also extends to extreme eastern California,
southern Nevada, southern Utah, and western New
Mexico.2–5 Growing interest in this species in the
pet trade and research arena has made acquisition of
baseline hematologic and biochemical values for these
animals relevant.
316
Animals and sample collection
Sixteen captive, clinically healthy adult Gila monsters
were used in this study. The animals had normal
behavior and water and food intake, and abnormalities were not detected on physical examination. Fourteen of the animals were wild-caught and comprised
11 males and 3 females captured in Arizona and maintained by one of the authors (D.D.); 2 were captivebred and were males owned by another author
c 2011 American Society for Veterinary Clinical Pathology
Vet Clin Pathol 40/3 (2011) 316–323 Cooper-Bailey et al
(K.C.B.). Using manual restraint, approximately 1 mL
of blood was obtained from the ventral tail vein of each
animal using a 23 G needle and 1 mL syringe. Each
sample was divided 3 ways: one drop was used to prepare 2 direct blood smears that were allowed to air dry
and 0.4–0.5 mL were placed in both a lithium heparin
and EDTA tube (Microtainer, Becton Dickinson,
Franklin Lakes, NJ, USA). Plasma was collected from
the lithium heparin samples within 10 minutes of collection following centrifugation at 1400g for 5 minutes. Plasma was then transferred to individual 1.5 mL
microcentrifuge tubes (SealRite, USA Scientific Inc.,
Ocala, FL, USA). EDTA samples were thoroughly
mixed by inversion. One blood smear and the plasma
and EDTA samples were sent by overnight courier to a
commercial diagnostic laboratory (Avian and Exotic
Animal Clinical Pathology Laboratories, Wilmington,
OH, USA) within 12–24 hours after collection.
Sample analysis
Samples were processed immediately upon receipt,
and a complete ‘‘reptile analysis,’’ including a CBC,
plasma biochemical analysis, and plasma electrophoresis, was performed. PCV was determined by centrifugation of the EDTA sample using a microhematocrit
centrifuge, and the buffy coat was examined for
microfilaria under low magnification ( 100) on a glass
slide. Hemoglobin concentration was determined by
an automated procedure using a hemoglobinometer
(Coulter Electronics, Hialeah, FL, USA). Erythrocyte
counts were performed manually using a hemocytometer (American Optical, Scientific Instrument Division, Buffalo, NY, USA), and WBC counts were
performed by the direct method with Natt and Herrick’s solution.6 Blood smears were stained with
Wright–Giemsa stain (Wescor 7120 Aerospray Hematology Stainer, Wescor Inc., Logan, UT, USA). A minimum of 100 sequential WBCs were counted from each
sample for determination of leukocyte differential values. All differential counts were performed by the
same individual (R.L.) and then reviewed by a second
individual (medical technician trained in reptilian
blood evaluation). For wild-caught animals (n = 14),
erythrocyte morphologic and morphometric characteristics (total cell area, area of the nucleus, cell length,
and width) were determined for cells in 10 random
fields on the Wright–Giemsa stained smears (K.Z.)
using a combination of software programs (ImageJ, NIH,
Bethesda, MD, USA; NIS Elements Laboratory Imaging
Software, Nikon Instruments Inc., Melville, NY, USA).7
Cytochemical analysis of WBCs for peroxidase,
chloroacetate esterase (CAE), naphthyl butyrate est-
Hematology and biochemistry of Gila monsters
erase (NBE), Sudan black B (SBB), and leukocyte
alkaline phosphatase (LAP) staining was qualitatively
assessed (R.E.R.) on blood smears from each animal
(n = 16) using standard cytochemical techniques.8–14
Immunocytochemical analysis for expression of B lymphocyte antigen (BLA.36, mouse monoclonal antibody, Novacastra, New Castle Upon Tyne, UK), CD3
(rabbit polyclonal antibody, Dako, Carpinteria, CA,
USA), and CD79a (mouse monoclonal antibody, Dako)
was also performed using additional blood smears;
however, these reagents have not been validated for
use in this species, and immunoreactivity was negative
in all cells (data not shown).
Plasma biochemical analysis was performed using
an automated chemistry analyzer (Cobas, Roche Diagnostics Limited, West Sussex, England, UK). Plasma in
a volume of 10 mL was analyzed by agarose gel electrophoresis using a semi-automated Sebia Hydrasys system (Sebia Inc., Norcross, GA, USA).
Statistical analysis
Numeric data from the CBC, plasma biochemical, and
electrophoretic data were analyzed for normality and
descriptive statistics determined using commercial statistical software (Minitab 15, Minitab Inc., State College, PA, USA).
Results
The CBC (Table 1) and RBC morphometric data
(Table 2) for all 16 Gila monsters and 14 wild-caught
animals, respectively, were normally distributed except for WBC counts and absolute heterophil and
azurophil counts. PCVs ranged from 22% to 50%.
Erythrocytes were oval, measured approximately
18 11 mm, had a typical orange cytoplasm, and had
central, round, homogenous, pale basophilic nuclei
(Figure 1A). Slight anisocytosis and polychromasia
were noted among the erythrocytes. Five different leukocyte types were noted in this study: heterophils,
lymphocytes, basophils, azurophils, and monocytes
(Figure 1B–F) in order of decreasing number. Eosinophils were not identified in any of the samples. Heterophils measured approximately 12 mm in diameter,
contained numerous, distinct, oval to sharp-pointed,
red-orange, cytoplasmic granules, and an oval, lightly
basophilic eccentric nucleus (Figure 1B). The median
heterophil percentage was 45% with several individuals, irrespective of the presence of hemoparasites, having 4 50% heterophils. Most of the lymphocytes were
large lymphocytes, measuring approximately 7 mm in
diameter. These cells were round and slightly larger
Vet Clin Pathol 40/3 (2011) 316–323 317
Cooper-Bailey et al
Table 1. Hematologic data from 16 Gila monsters (Heloderma suspectum).
Variable
Mean
SD
Minimum
Q1
Median
Q3
Maximum
RBC ( 106/mL)
PCV (%)
Hemoglobin (g/dL)
MCV (fL)
MCHC (g/dL)
WBC ( 103/mL)
Heterophils ( 103/mL)
Lymphocytes ( 103/mL)
Basophils ( 103/mL)
Azurophils ( 103/mL)
Monocytes ( 103/mL)
Eosinophils ( 103/mL)
Total solids (g/dL)
0.50
37
7.4
812
20.9
4.72
2.17
1.54
0.57
0.38
0.07
0.00
6.3
0.13
8
0.9
370
5.3
0.84
0.61
0.80
0.23
0.27
0.08
0.00
1.0
0.22
22
6.0
415
14.0
3.30
1.35
0.58
0.21
0.00
0.00
0.00
5.0
0.42
32
6.5
537
17.0
4.20
1.69
0.73
0.44
0.14
0.00
0.00
5.5
0.53
37
7.5
648
21.0
4.60
2.08
1.61
0.54
0.38
0.00
0.00
6.0
0.61
44
8.0
978
23.0
5.20
2.57
2.01
0.64
0.47
0.16
0.00
7.0
0.67
50
9.5
1773
36.0
6.40
3.31
3.39
1.05
1.14
0.19
0.00
8.4
Distribution of data not normal.
Q, quartile.
than thrombocytes, contained round nuclei with
smudged chromatin, and had a faint rim of basophilic
cytoplasm (Figure 1C). Basophils were distinct with
their numerous dark purple granules partially obscuring
an oval eccentric nucleus and measured approximately
10 mm in diameter (Figure 1D). Azurophils measured
approximately 10 mm in diameter, contained faint azurophilic granules with blue-grey cytoplasm, which lacked
vacuoles, and had slightly indented basophilic nuclei
(Figure 1E). Monocytes were similar to azurophils in
size, color, and nuclear shape, with the only difference
being the lack of azurophilic granules (Figure 1F).
Thrombocytes were approximately 5 mm in diameter
and round with dark, basophilic, homogenous nuclei,
and scant cytoplasm ranging from almost colorless to
pale grey-blue containing a few indistinct vacuoles
and basophilic granules (Figure 1G). Platelet numbers
were not evaluated. Four of the animals were parasitized
with an unidentified filarial parasite (Figure 1H).
Leukocytes had variable staining patterns for peroxidase, SBB, CAE, and NBE (Table 3, Figure 2).
Heterophils and azurophils/monocytes had similar
positive staining for peroxidase and SBB. In heterophils peroxidase staining was weak and granular. In
monocytes SBB staining was stronger and more globular. Basophils had trace positivity for SBB between
granules, which were negative. Lymphocytes and
thrombocytes did not stain with any of the cytochemical stains tested except for rare focal positivity for CAE
in lymphocytes. Heterophils stained for both CAE and
NBE between the granules, and the cytoplasm of
azurophils/monocytes stained diffusely. None of the
leukocytes stained for LAP (Table 3).
Plasma biochemical (Table 4) and protein electrophoretic (Table 5) profiles were determined. Data for
biochemical analytes were normally distributed except
for phosphorus and bile acid concentrations and creatine kinase activity. For plasma electrophoretic values
only albumin and b-fractions were normally distributed. A representative electrophoretic tracing from
one of the study animals was compared with a tracing
from an animal (outside the study) with leukocytosis
resulting from retained eggs (Figure 3). The tracing
from the ill animal showed a relative increase in the a-,
b-, and g-globulin fractions as well as increased bridging of the b–g region.
Discussion
Baseline values for wild populations of threatened or
endangered species are valuable for reference in
Table 2. Morphometric analysis of erythrocytes from 14 wild-caught Gila monsters (Heloderma suspectum).
Measurement
Mean
SD
Minimum
Q1
Median
Q3
Maximum
Total area (mm2)
Area of nucleus (mm2)
Length (mm)
Width (mm)
150.57
22.40
17.83
10.57
21.20
3.24
1.30
1.05
103.77
15.00
14.80
7.61
136.45
19.98
16.97
9.93
149.13
22.32
17.86
10.57
164.03
24.82
18.72
11.33
197.35
31.54
21.77
13.08
Q, quartile.
318
Figure 1. Blood from a wild-caught captive Gila monster (Heloderma suspectum). Modified-Wright’s stain, bars = 10 mm. (A) Erythrocyte with hemoglobinized cytoplasm, elliptical shape, and oval central basophilic nucleus. (B) Heterophil with numerous distinct oval to pointed red-orange granules and a
basophilic oval eccentric nucleus. (C) Lymphocyte with a round nucleus, smudged chromatin, and faint basophilic cytoplasm. (D) Basophil with dark
purple granules obscuring an oval eccentric nucleus. (E) Azurophil with faint azurophilic granules, blue-grey cytoplasm lacking vacuoles, and a reniform
basophilic nucleus. (F) Monocyte with a nucleus similar to that of the azurophil, but with slightly more basophilic cytoplasm and lacking azurophilic
granules. (G) Thrombocyte with a round dense homogenous nucleus and scant colorless to pale grey-blue cytoplasm with a few indistinct vacuoles. (H)
Unidentified filarial parasite seen in 4 blood samples; larva were approximately 200 8 mm with a pointed anterior end and a rounded posterior end, no
apparent body cavity, and numerous 2-3 mm oval dark basophilic nuclei.
captive health and conservation programs. In addition,
with growing interest in the Gila monster as an ornamental, educational, and scientific study animal, baseline values for captive individuals are needed for the
veterinary community and its work with exotic species.11,15–18 Variations due to availability of resources,
diet, climatic changes, and activity are often found
between captive and wild populations of reptiles; this
trend was also suggested by our results. Further analysis of the data (not shown owing to low numbers of
animals in groups) also suggested a possible influence
of wild-caught versus captive-bred status on hematologic, biochemical, and protein electrophoretic data;
the need for independent reference intervals should be
considered.
Quantitative and morphologic characteristics of
blood cells are important in the assessment of reptilian
health, but may vary among species.19–24 In a previous
study, erythrocytes and leukocytes of the Mexican
beaded lizard, Heloderma horridum, the sister species of
the Gila monster, were described and served as a guide
for the current study.22 PCVs from this study were in
agreement with those reported as normal for reptiles,
20–40%.21,25,26 For leukocyctes, heterophils were
most numerous. It has been reported that heterophil
percentages can be as high as 40% of the overall leukocyte count for some normal reptilian species.21,25,27
In this study, Gila monsters had a higher median heterophil percentage than is typical of other
reptiles. The next most common leukocyte type was
Table 3. Cytochemical staining reactions of WBCs from Gila monsters (Heloderma suspectum).
Stain/reaction
Peroxidase
SBB
CAE
NBE
LAP
Heterophil
Lymphocyte
Basophil
Azurophil/monocyte
Thrombocyte
1 (Granular)
1 (Diffuse)
1
1
-
1 (Rare focal)
-
1
-
11 (Granular)
111 (Granular)
11 (Diffuse)
11 (Diffuse)
-
-
SBB, Sudan black B; CAE, chloroacetate esterase; NBE, naphthyl butyrate esterase; LAP, leukocyte alkaline phosphatase;
itive; 11, moderately positive; 111, strongly positive; , intergranular reaction.
Vet Clin Pathol 40/3 (2011) 316–323 , negative; 1, weakly pos-
319
Cooper-Bailey et al
Figure 2. Blood from Gila monsters (Heloderma suspectum) stained for chloroacetate esterase (A, B), peroxidase (C, D), naphthyl butyrate esterase (E,
F), and Sudan black B (G–I). h, heterophil; m, monocyte/azurophil; b, basophil; l, lymphocyte; t, thrombocyte. See text for details.
Table 4. Plasma biochemical data from 16 Gila monsters (Heloderma suspectum).
Analyte
Mean
SD
Minimum
Q1
Median
Q3
Maximum
Total protein (g/dL)
Glucose (mg/dL)
Phosphorus (mg/dL)
Sodium (mmol/L)
Potassium (mmol/L)w
Ionized calcium (mmol/L)
pH
Total calcium (mg/dL)
Creatine kinase (U/L)w
Aspartate aminotransferase (U/L)
Bile acids (mmol/L)w
Urea (mg/dL)
Uric acid (mg/dL)
6.3
48.3
3.4
144
3.9
1.26
7.66
12.2
600
42
16.2
15
16.8
0.5
24.2
1.8
3
0.5
0.10
0.09
0.8
457
13
13.3
6
4.2
5.4
4.0
1.1
140
2.8
1.09
7.48
10.2
144
20
2.6
6
9.8
6.1
36.3
2.3
141
3.5
1.19
7.60
11.7
253
30
7.0
13
14.1
6.3
48.0
2.8
143
4.0
1.23
7.67
12.2
464
42
12.4
14
17.2
6.6
58.5
4.0
147
4.230
1.32
7.74
13.0
830
51
21.7
17
19.0
6.9
109.0
8.60
151
4.6
1.50
7.79
13.4
1812
66
55.1
30
24.7
Total protein measured by biuret reaction; glucose, hexokinase; phosphorus, phosphomolybdate-UV; sodium, potassium, ionized calcium, pH, direct
undiluted measurement by ion selective electrodes; total calcium, arsenazo III dye; creatine kinase, UV-kinetic; aspartate aminotrasferase, IFCC UVkinetic; bile acids, 3a-HSD; and urea and uric acid, kinetic spectrophotmetry.
w
Q, quartile.
320
Table 5. Plasma protein electrophoretic values from 16 Gila monsters
(Heloderma suspectum).
Analyte
Mean
Albumin (g/dL)
a-1 (g/dL)
a-2 (g/dL)
b (g/dL)
g (g/dL)
Albumin (%)
a-1 (%)
a-2 (%)
b (%)
g (%)
2.61
2.09
0.59
0.58
0.33
42.1
33.8
9.4
9.33
5.4
SD Minimum
0.28
0.27
0.10
0.10
0.13
2.2
3.6
1.2
1.2
1.9
2.14
1.48
0.44
0.41
0.18
38.7
23.2
7.3
7.9
2.7
Q1
Median
2.36 2.60
1.95 2.07
0.50 0.59
0.51 0.54
0.25 0.33
40.1 42.0
32.6 33.7
8.7
9.3
8.4
9.0
4.3
4.6
Q3
Maximum
2.79
2.25
0.67
0.66
0.40
43.5
36.6
10.4
10.1
6.5
3.23
2.60
0.76
0.77
0.68
46.5
38.8
11.3
12.8
10.6
Q, quartile.
lymphocytes. Although both small and large lymphocytes have been reported from various species of reptiles, the majority of the lymphocytes observed in this
study were large. The wide range in lymphocyte percentages is common in many species of reptiles and
thus was considered normal.15,21,22,25 Basophils, the
next most common leukocyte, were similar in percentage to those described for other reptiles.20,21,26 The low
percentages of azurophils and monocytes and absence
of eosinophils in these study animals are in agreement
with reports in the reptilian literature.21,25
Heterophils from chelonians (turtles and tortoises)
and crocodilians (alligators, caimans, and crocodiles)
have been uniformly reported as being peroxidasenegative.9,12,28,29 Similarly, heterophils and azurophils
from some Squamata (lizards and snakes, eg, the eastern diamondback rattlesnake, Crotalus adamanteus)
have also been reported as being peroxidase-negative,
whereas heterophils and azurophils from other squamates, eg, the green iguana, Iguana iguana, have been
Figure 3. Plasma protein electrophoretograms from (A) a healthy wild
bred-captive Gila monster (Heloderma suspectum) with the albumin fraction representing the largest single fraction and approximately equal to
the combined globulin fractions and (B) a captive-bred female with retained eggs showing reduced albumin:globulin ratio. a-Globulins represented the highest single fraction in this animal, and a relative increase in
bridging between the b and g fractions was present.
reported as being peroxidase-positive.8,11,12,28,30,31 In
this study, both heterophils and azurophils of the Gila
monster demonstrated varying degrees of peroxidase
staining in the cytoplasm. However, staining in azurophils/monocytes was much stronger than in heterophils,
which is in contrast to reports in other lizards in which
peroxidase staining was negative in azurophils.11 SBB
staining was also distinct in our study animals as
azurophils/monocytes were strongly positive and
heterophils were weakly positive. In green iguanas,
heterophils were found to be strongly positive for SBB
and azurophil/monocytes were negative.11 Also unique
among most species of reptiles was the positive cytoplasmic staining by SBB within the basophils of Gila
monsters.8,10,11,21,28 In addition, in contrast to reports in
green iguanas where both heterophils and azurophils
were found to be CAE negative, in this study CAE positivity was seen in both heterophils and azuruphils, as
well as in some lymphocytes with focal staining.11
Most biochemical values were consistent with those
reported for the congeneric Mexican beaded lizard, as
well as with those reported for other reptilian species.22,23,25 The only noteworthy differences between
the 2 species of Heloderma were the plasma urea and uric
acid concentrations, which were higher for the Gila
monsters in this study.22 High concentrations of these
analytes may suggest renal disease, prerenal azotemia,
or high dietary intake of protein. Prerenal azotemia from
dehydration was considered less likely because the individuals with higher PCVs or total protein concentrations
were not those with the higher urea and uric acid concentrations. These concentrations were most likely due
to dietary intake as captive reptiles are often fed high
protein diets consisting of whole rodents at increased
frequencies compared with wild-caught animals in the
beaded lizard study.
Data for protein electrophoresis have not been
reported for the genus Heloderma. In this study the
albumin fraction represented the largest single fraction
and was approximately equal to the combined globulin
fractions. Interestingly, the relative percentage of a-1
and -2 globulins was higher than is typical for most
mammalian species.32 Another interesting observation
was the prominence of the a-peak in these animals.
Similar prominent a-peaks have been reported in people and species of Salamandra and Iguana and result
from the presence of bisalbuminemia.33,34 The cause of
the prominent a-peak in the Gila monsters is not clear.
It is important to consider that electrophoresis was performed using plasma, and fibrinogen may have been a
contributing factor in the presence of the peak in the
late b and early g fractions. The use of electrophoretic
data is becoming an increasingly important modality
Vet Clin Pathol 40/3 (2011) 316–323 321
Cooper-Bailey et al
for diagnosis of acute and chronic inflammatory diseases, and values are being determined for different
reptilian species. However, comparisons are difficult
owing to species variation.7,35,36 During the course of
this study, electrophoretic data were collected from a
gravid female Gila monster with retained eggs, and
tracings from the healthy animals could be used as a
baseline for comparison. Reduced albumin:globulin
ratio and an increased globulin fraction with aglobulins representing the greatest single protein fraction were changes typical of those found in association
with chronic inflammation in mammalian species.32,37
Microfilaria have been reported in reptiles and
often do not cause overt clinical disease.23,25 Freeranging Gila monsters and 19% of wild-caught Mexican beaded lizards have been reported to be parasitized
with an unidentified filarial parasite with no apparent
anemia or altered blood values.22,38 In this study, all of
the filaria-parasitized animals were asymptomatic and
it is likely that the parasitism did not affect results.
In conclusion, this study provides preliminary reference values for clinically healthy Gila monsters that
will aid veterinarians in clinical evaluation and monitoring of both healthy and ill Gila monsters. Further
work is needed to determine whether significant
differences in values between wild-caught and captive-bred animals are present.
Acknowledgments
The authors gratefully acknowledge Philip Bailey (Blacksburg, VA) and Christian Wright and Karla Moeller from the
School of Life Sciences, Arizona State University, Tempe,
AZ for their time and assistance in handling and sampling of
the animals and personnel from the Avian and Exotic Clinical Pathology Laboratories (Wilmington, OH) for providing
the electrophoretic data.
Disclosure: The authors have indicated that they have
no affiliations or financial involvement with any organization or entity with a financial interest in, or in financial
competition with, the subject matter or materials discussed
in this article.
References
1. Fry BG, Vidal N, Norman JA, et al. Early evolution of
venom production in lizards and snakes. Nature.
2006;439:584–588.
2. Beaman K, Beck D, McGurty BM. The Beaded Lizard
(Heloderma horridum) and Gila Monster (Heloderma
suspectum): A Bibliography of the Family Helodermatidae.
Washington, D.C.: National Museum of Natural
History, Smithsonian Herpetological Information
Service; 2006.
322
3. Alden P, Society NA. National Audubon Society Field
Guide to the Southwestern States. 1st ed. New York, NY:
Knopf/Distributed by Random House; 1999.
4. Beck DD. Biology of Gila Monsters and Beaded Lizards.
Berkeley, CA: University of California Press; 2005.
5. Degenhardt WG, Painter CW, Price AH. Amphibians and
Reptiles of New Mexico. 1st ed. Albuquerque, NM:
University of New Mexico Press; 1996:247–251.
6. Natt M, Herrick C. A new blood diluent for counting
the erythrocytes and leucocytes of the chicken. Poult
Sci. 1952;31:735–738.
7. Zaias J, Norton T, Fickel A, Spratt J, Altman NH, Cary C.
Biochemical and hematologic values for 18 clinically
healthy radiated tortoises (Geochelone radiata) on St
Catherines Island, Georgia. Vet Clin Pathol. 2006;
35:321–325.
8. Alleman AR, Jacobson ER, Raskin RE. Morphologic,
cytochemical staining, and ultrastructural
characteristics of blood cells from eastern diamondback
rattlesnakes (Crotalus adamanteus). Am J Vet Res. 1999;
60:507–514.
9. Raskin RE. Cytochemical Staining. In: Weiss DJ,
Wardrop KJ, eds. Schalm’s Veterinary Hematology. 6th ed.
Ames, IA: Blackwell Publishing; 2010:1141–1156.
10. Allender MC, Fry MM. Amphibian hematology. Vet Clin
North Am Exot Anim Pract. 2008;11:463–480, vi.
11. Harr KE, Alleman AR, Dennis PM, et al. Morphologic
and cytochemical characteristics of blood cells and
hematologic and plasma biochemical reference ranges
in green iguanas. J Am Vet Med Assoc. 2001;218:
915–921.
12. Mateo MR, Roberts ED, Enright FM. Morphologic,
cytochemical, and functional studies of peripheral
blood cells of young healthy American alligators
(Alligator mississippiensis). Am J Vet Res. 1984;45:
1046–1053.
13. Pettigrew NM. Techniques in immunocytochemistry.
Application to diagnostic pathology. Arch Pathol Lab
Med. 1989;113:641–644.
14. Dinges HP, Wirnsberger G, Hofler H.
Immunocytochemistry in cytology. Comparative
evaluation of different techniques. Anal Quant Cytol
Histol. 1989;11:22–32.
15. Dutton CJ, Taylor P. A comparison between pre- and
posthibernation morphometry, hematology, and blood
chemistry in viperid snakes. J Zoo Wildl Med. 2003;
34:53–58.
16. Eliman MM. Hematology and plasma chemistry of the
inland bearded dragon, Pogona vitticeps. Reptil Amphib
Vet. 1997;7:10–12.
17. ISIS. Physiological Data Reference Values for the Bearded
Dragon (Pogona vitticeps). Apple Valley, MN:
International Species Information System; 1999.
18. Lamirande EW, Bratthauer AD, Fischer DC, Nichols
DK. Reference hematologic and plasma chemistry
values of brown tree snakes (Boiga irregularis). J Zoo
Wildl Med. 1999;30:516–520.
19. Canfield PJ. Comparative cell morphology in the
peripheral blood film from exotic and native animals.
Aust Vet J. 1998;76:793–800.
20. Kelly JW, Boseila AW, Cavazos LF, Feagans WM.
Hematology of the horned lizard, especially blood
basophils and tissue mast cells. Acta Haematol. 1961;
26:378–384.
21. Sykes JMT, Klaphake E. Reptile hematology. Vet Clin
North Am Exot Anim Pract. 2008;11:481–500.
22. Espinosa-Aviles D, Salomon-Soto VM, MoralesMartinez S. Hematology, blood chemistry, and
bacteriology of the free-ranging Mexican beaded lizard
(Heloderma horridum). J Zoo Wildl Med. 2008;39:21–27.
23. Campbell TW. Clinical pathology. In: Mader DR, ed.
Reptile Medicine and Surgery. Philadelphia, PA: W. B.
Saunders Co; 1996:248–257.
24. Schumacher JL. Lacertilia (lizards, skinks and geckos)
and amphisbaenids (worm lizards). In: Fowler ME,
Miller RE, eds. Zoo and Wild Animal Medicine. 5th ed. St.
Louis, MO: Saunders; 2003:73–81.
25. Thrall MA. Hematology of reptiles. In: Thrall MA, ed.
Veterinary Hematology and Clinical Chemistry.
Philadelphia, PA: Lippincott Williams & Wilkins;
2004:259–290.
26. Reagan WJ, Irizarry Rovira AR, DeNicola DB. Veterinary
Hematology: Atlas of Common Domestic and Non-Domestic
Species. 2nd ed. Ames, IA: Blackwell Publishing;
2008:85–96.
27. Irizarry Rovira AR. Hematology of Reptiles. In: Weiss
DJ, Wardrop KJ, eds Schalm’s Veterinary Hematology. 6th
ed. Ames, IA: Blackwell Publishing; 2010:1004–1012.
28. Alleman AR, Jacobson ER, Raskin RE. Morphologic
and cytochemical characteristics of blood cells from the
desert tortoise (Gopherus agassizii). Am J Vet Res.
1992;53:1645–1651.
29. Strik N, Alleman AR, Harr KE. Circulating
Inflammatory Cells. In: Jacobson ER, ed. Infectious
Diseases and Pathology of Reptiles: Color Atlas and Text.
Boca Raton, FL: CRC/Taylor & Francis; 2007:167–218.
30. Bounous DI, Dotson TK, Brooks RL, Ramsay C.
Cytochemical staining and ultrastructural
characteristics of peripheral blood leucocytes from the
yellow rat snake (Elaphe obsoleta quadrivitatta). Comp
Haematol Int. 1996;6:86–91.
31. Ramos-Vara JA, Avery AC, Avery PR. Advanced
diagnostic techniques. In: Raskin R, Meyer DJ, eds.
Canine and Feline Cytology: A Color atlas and Interpretation
Guide. 2nd ed. St. Louis, MO: Saunders Elsevier;
2010:395–437.
32. Jain NC. The Plasma Proteins, Dysproteinemias, and
Immune Deficiency Disorders. Essentials of Veterinary
Hematology. Philadelphia, PA: Lea & Febiger;
1993:349–380.
33. Gimenez M, Saco Y, Pato R, Busquets A, Martorell JM,
Bassols A. Plasma protein electrophoresis of Trachemys
scripta and Iguana iguana. Vet Clin Pathol. 2010;39:
227–235.
34. Rossi S, Bertazzolo W, Paltrinieri S, Giordano A.
Cellulose acetate electrophoresis of canine plasma after
fibrinogen precipitation by ethanol. Vet Clin Pathol.
2008;37:422–428.
35. Deem SL, Dierenfeld ES, Sounguet GP, et al. Blood
values in free-ranging nesting leatherback sea turtles
(Dermochelys coriacea) on the coast of the Republic of
Gabon. J Zoo Wildl Med. 2006;37:464–471.
36. Machado CC, Silva LF, Ramos PR, Takahira RK.
Seasonal influence on hematologic values and
hemoglobin electrophoresis in Brazilian Boa constrictor
amarali. J Zoo Wildl Med. 2006;37:487–491.
37. Evans EW, Duncan JR. Proteins, lipids and
carbohydrates. In: Latimer KS, Mahaffey EA, Prasse
KW, eds. Duncan & Prasse’s Veterinary Laboratory
Medicine: Clinical Pathology. 4th ed. Ames, IA: Iowa State
Press; 2003:162–192.
38. Mahrt JL. Hematozoa of lizards from Southeastern
Arizona and Isla San-Pedro-Nolasco, Gulf of California,
Mexico. J Parasitol. 1979;65:972–975.
Vet Clin Pathol 40/3 (2011) 316–323 323

Hematology, blood chemistry and plasma

Transcription

Similar documents

Gila Lizard.indd

the ballad of great garloo`s revenge - all

Created by Devon Meekis - United Way of Thunder Bay

Plasma Thawer - kizlonmedical.com

`Frightening` Frankie, `Dangerous` Drac, and (Yeah, Right!) `Weirdo

Papier-Mâché Monsters EBLAD

plasmabit - Norsk olje og gass

Sea Monsters

BioLife Plasma Center in American Fork “Apply Now”

5. Diet - ZooCentral