COMPARISON OF BLOOD VALUES IN FORAGING, NESTING, AND
STRANDED LOGGERHEAD TURTLES (CARETTA CARETTA) ALONG
THE COAST OF GEORGIA, USA
Sharon L. Deem,
1,11,13
Terry M. Norton,
2,3
Mark Mitchell,
4
Al Segars,
5
A. Rick Alleman,
6
Carolyn Cray,
7
RobertH. Poppenga,
8,12
Mark Dodd,
9
and William B. Karesh
10
1
Department of Animal Health, Smithsonian National Zoological Park, 3001 Connecticut Ave. NW, Washington, D.C.
20008, USA
2
St. Catherines Island Center, 182 Camellia Road, Midway, Georgia 31320, USA
3
Georgia Sea Turtle Center, 214 Stable Road, Jekyll Island, Georgia 31527, USA
4
Department of Veterinary Clinical Medicine, 270 SAC MC-004, 1008 W Hazelwood Drive, Urbana, Illinois 61802, USA
5
South Carolina Department of Natural Resources, 32 Fiddler Drive, Beaufort, South Carolina 29902, USA
6
Department of Physiological Sciences, College of Veterinary Medicine, PO Box 100103, University of Florida, College
of Veterinary Medicine, Gainesville, Florida 32610, USA
7
Department of Pathology, University of Miami School of Medicine, 1600 NW 10th Ave., University of Miami, Miami,
Florida 33101, USA
8
University of Pennsylvania, School of Veterinary Medicine, New Bolton Center, 382 W Street Road, Kennett Square,
Pennsylvania 19348, USA
9
Georgia Department of Natural Resources, Non-game Wildlife-Natural Heritage Section, One Conservation Way, Suite
310, Brunswick, Georgia 31520-8687, USA
10
Field Veterinary Program, Wildlife Conservation Society, 2300 Southern Boulevard, New York, New York 10460, USA
11
Present Address: WildCare Institute, Saint Louis Zoo, One Government Drive, Saint Louis, Missouri, 63110-1396,
USA
12
Present Address: California Animal Health and Food Safety Toxicology Laboratory, School of Veterinary Medicine,
University of California, West Health Sciences Drive, Davis, California 95616, USA
13
Corresponding author (email: deem@stlzoo.org)
ABSTRACT: The health status of 83 loggerhead sea turtles (Caretta caretta; 39 foraging, 31 nesting,
and 13 stranded turtles) was analyzed using physical examinations, hematology, plasma
biochemistry, plasma protein electrophoresis, and toxicologic parameters. Significant differences
were noted in a number of health parameters between turtles exhibiting each of these behaviors.
On physical examinations, stranded turtles had the highest prevalence of heavy carapace epibiont
loads, miscellaneous abnormalities, emaciation, and weakness. Differences in hematologic values
included a lower packed cell volume, higher number of lymphocytes, and lower number of
monocytes in stranded turtles; lower white blood cell counts in foraging turtles; and significant
differences in total solid values among turtles exhibiting all behaviors with the lowest values in
stranded turtles and the highest values in nesting turtles. Differences in plasma biochemistry
values included the highest uric acid, creatine kinase, and CO
2
values in stranded turtles; the
highest glucose and potassium values in foraging turtles; and the highest cholesterol and
triglyceride values, and lowest alanine aminotransferase, in nesting turtles. Differences in total
protein, albumin, and globulin were found using plasma biochemistry values, with lowest values in
stranded turtles and highest values in nesting females, whereas differences in blood urea nitrogen
between turtles included the lowest values in nesting turtles and the highest in foraging turtles.
Plasma organochlorine and polychlorinated biphenyl levels were below their limits of
quantification in the 39 foraging, 11 nesting, and three stranded turtles tested. A statistically
significant difference was noted in the level of whole blood mercury between the 23 foraging and
12 nesting turtles tested. There was no difference in arsenic or lead levels between turtles
exhibiting any of the three behaviors. Although a few limitations exist with the present study and
include unknown ambient temperatures, turtle handling times that varied from 15 min to 53 min
per turtle, and the use of a different laboratory for processing complete blood counts and plasma
biochemistries in stranded versus foraging and nesting turtles, we provide baseline blood values for
two cohorts (foraging and nesting) of loggerhead sea turtles on the coast of Georgia. Additionally,
we demonstrate significant differences in clinical findings and blood parameters between foraging,
nesting, and stranded loggerhead turtles in the region.
Key words: Caretta caretta, hematology, loggerhead turtles, plasma biochemistry, plasma
protein electrophoresis, toxicants.
Journal of Wildlife Diseases, 45(1), 2009, pp. 41?56
# Wildlife Disease Association 2009
41
INTRODUCTION
Loggerhead sea turtles (Caretta caretta)
are the most common sea turtle species
nesting on the coast of Georgia (Norton,
2005). In addition to loggerheads, four
other sea turtle species including green
(Chelonia mydas), leatherback (Dermo-
chelys coriacea), hawksbill (Eretmochelys
imbricata), and Kemp?s ridley (Lepido-
chelys kempi) use coastal Georgia waters
for food, shelter, occasional nesting, and as
a travel corridor (Norton, 2005). Conser-
vation threats to sea turtles in Georgia are
primarily associated with beachfront de-
velopment, pollution from chemical tox-
ins, ingestion of marine debris and drown-
ing and traumatic injuries from incidental
entanglement related to recreational and
commercial fisheries activities (Norton,
2005). Additionally, diseases, including
the debilitated loggerhead sea turtle syn-
drome, are direct threats to loggerhead
conservation (George, 1997; Norton et al.,
2005; Jacobson et al., 2006).
In addition to monitoring population
demographics and causes of mortality,
baseline blood values can be used as one
indicator of population health. Life stages
(e.g., foraging, nesting), age, sex, nutri-
tional status, pathogen, parasite and toxin
exposure, and environmental conditions
all impact physiologic status and influence
baseline blood parameters in sea turtles
(Whitaker and Krum, 1999; Herbst and
Jacobson, 2003). Therefore, the establish-
ment of baseline values for different
cohorts within a population must be
obtained. These baseline values are in-
valuable for providing an indication of the
overall health of a wild population, for
measuring the health of a population over
time, for comparing the health of popula-
tions, and for use as prognostic indicators
for individual health assessments of
stranded turtles.
The objective of this study was to
compare health indices, including physical
examination findings, hematology, plasma
biochemistry values, plasma protein elec-
trophoresis, and toxicologic parameters of
foraging, nesting, and stranded loggerhead
sea turtles on the coast of Georgia.
MATERIALS AND METHODS
Study period and site
This study was carried out between May and
September during the years 2000 to 2004.
Foraging turtles (n539) were captured by
trawl in waters off the coast of northeast
Florida and Georgia in conjunction with
ongoing studies of sea turtle population
biology. Foraging turtles were collected by
fishery independent trawlers operating in
near-shore waters between Savannah, Geor-
gia, USA (32u59N, 81u59W) and St. Augustine,
Florida, USA (29u509N, 81u159W) in 4.5?12.2-
m-deep water. (Maier et al., 2004). Trawlers
used 20-m four-seam nets with 20-cm mesh,
without turtle excluder devices and trawl
duration was limited to 30 min, as previously
described (Maier et al., 2004). Nesting females
were approached on the beach after they
completed egg laying and were returning to
the ocean. Nesting females were sampled on
Blackbeard Island, Georgia, USA (31u309N,
81u259W) (n527); Cumberland Island, Geor-
gia, USA (30u519N, 81u269W) (n51); and
Jekyll Island, Georgia, USA (31u49N,
81u259W) (n53). Live stranded turtles
(n513) were found all along the Georgia coast
and transported to the Georgia Department of
Natural Resources for their initial evaluation.
Sample and data collection and analyses
A complete physical examination was per-
formed and abnormalities categorized into
heavy epibiont load, flipper amputations,
miscellaneous abnormalities (e.g., propeller
wound, shark bite, fishing hook, neurologic
signs), emaciation, and weakness. Epibiont
loads were based on epibiont presence on the
carapace categorized using the ordinal scale of
1to3(15mild [,20 epibionts]; 25moderate
[20?50 epibionts]; 35heavy [.50 epibionts]).
Curved carapace length (CCL), notch to tip,
was measured using a nylon tape measure.
Foraging and stranded turtles were weighed
using hanging spring scales. Nesting females
were not weighed. A turtle was classified as
emaciated based on a body weight that was
less than expected for known carapace length
(in those turtles in which both parameters
were collected), and a sunken appearance to
axillae, flank, and/or plastron regions. A turtle
was classified as weak if it displayed minimal
response to human handling with minimal to
42 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
no head and flipper movements. Age (adult
versus subadult) of foraging and stranded
turtles was determined based on CCL; with
adults $87 cm and subadults ,87 cm. Age
categories were established using the smallest
nesting adult female in our study. Sex was
determined using tail length (e.g., long tail in
adult males) in adult stranded turtles that
survived and visualization of the gonad at
necropsy in stranded turtles that did not
survive. In foraging animals, plasma testoster-
one levels were measured for sex determina-
tion using radioimmunoassay as previously
described (Owens et al., 1978). Complete
postmortem examinations (gross and micro-
scopic examination) were performed on all
stranded turtles that died (n56).
Approximately 35 ml of blood was collected
from the dorsal cervical sinus using a 20 gauge
or 22 gauge 3.8-cm needle and dispensed
directly into large (10-ml) vacutainer tubes
containing either lithium heparin (Corvac,
Sherwood Medical, Saint Louis, Missouri,
USA), for hematology and plasma biochemis-
try values, or buffered citrate sodium (Becton-
Dickinson Diagnostics, Pre-Analytical Sys-
tems, Franklin Lakes, New Jersey, USA) for
heavy metal testing (Owens and Ruiz, 1980).
Tubes were kept in a cooler with ice packs
until processed.
Initial processing of blood samples occurred
in the field within 4 hr (range 15 min to 4 hr)
of blood collection for trawl-caught turtles,
nesting females, and stranded turtles. Thin
blood smears were made within 30 min of
collection and then fixed in the field with 99%
methanol. Packed cell volumes (PCV) were
determined using a portable 12-V centrifuge
(Mobilespin, Vulcan Technologies, Grand-
view, Missouri, USA) and plasma total solids
were measured using a handheld refractome-
ter (Schulco, Toledo, Ohio, USA) tempera-
ture-calibrated at the site. White blood cell
(WBC) counts were performed using the BD
UnopetteH brand test for manual eosinophil
counts (Catalog 365877, Becton-Dickinson
Diagnostics, Pre-analytical Systems,). Red
blood cell (RBC) counts were made using
the BD UnopetteH brand test for manual rbc
counts (Catalog 365850/365851, Becton-Dick-
inson Diagnostics, Pre-analytical Systems).
The remaining heparinized blood was centri-
fuged for 10 min and plasma was then
pipetted into cryotubes (Corning Incorporat-
ed, Corning, New York, USA). Plasma was
kept frozen (270 C) and transported on dry
ice (n570) to laboratories for diagnostic
testing, or shipped immediately on wet ice
for stranded turtles (n513). Whole blood
collected in buffered citrate sodium tubes
was placed in cryotubes (Corning Incorporat-
ed) and kept frozen (270 C) and transported
on dry ice to the laboratory. All laboratory
diagnostics were performed within 6 mo of
blood collection.
For foraging and nesting loggerheads, blood
films fixed in methanol were stained with
Wright-Giemsa, at the University of Florida,
for evaluation of circulating cell morphology,
estimation of leukocyte numbers, and differ-
ential leukocyte counts. A minimum of 200
leukocytes were counted for differential leu-
kocyte determinations. Leukocytes were cate-
gorized into one of six groups: monocytes,
heterophils, lymphocytes, eosinophils, baso-
phils, and azurophils. Identification of blood
cell types was based on previously described
nomenclature (Hawkey and Dennett, 1989).
Red blood cells were evaluated for hemopar-
asite identification. Additionally, total white
blood cell counts were estimated from blood
films by multiplying the average number of
leukocytes observed per microscopic field
times the objective power squared (Harvey,
2001). White blood cell estimates and differ-
ential cell counts were performed on blood
films from stranded turtles by a commercial
laboratory (Antech Diagnostic Laboratories,
Memphis, Tennessee, USA) as part of their
complete blood count (CBC) (Antech?s Com-
prehensive Reptile Profile-AE 160). For the
stranded turtles, the blood smear was re-
viewed for the WBC estimate using a 403 lens
with the average of the WBCs in 10 consec-
utive fields multiplied by 1,000 to obtain the
WBC estimate.
For foraging and nesting loggerheads, sam-
ples for plasma biochemistry were processed
on a dry slide chemistry analyzer (Kodak 750 X
R, Ortho Clinical Diagnostics, Rochester, New
York, USA) at the University of Miami. The
biochemical panel included albumin, alanine
aminotransferase (ALT), amylase, aspartate
aminotransferase (AST), blood urea nitrogen
(BUN), calcium, chloride, cholesterol, CO
2
,
creatine kinase (CK), creatinine, gamma glu-
tamyl transferase (GGT), globulin, glucose,
lactate dehydrogenase, lipase, phosphorous,
potassium, sodium, total protein, triglyceride,
and uric acid (UA). Plasma biochemistries for
stranded turtles were performed by a com-
mercial laboratory (Antech Diagnostic Labo-
ratories) as part of their chemistry profiles
using a Hitachi 747-100.
Plasma electrophoresis was performed at
the University of Miami using SPEP-II
agarose gels and the Beckman paragon elec-
trophoresis system (Beckman-Coulter Corpo-
ration, Brea, California, USA). The gels were
run according to manufacturer?s instructions
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 43
as described previously (Cray and Tatum,
1998). The percentage of protein fractions
was quantitated by laser densitometry and
then each fraction value was calculated by
multiplying the percentage of the fraction by
the total protein value determined using the
dry chemistry analyzer.
Plasma was screened at the University of
Pennsylvania for the presence of organochlo-
rine (OC) insecticides (aldrin, alpha-BHC,
beta-BHC, alpha-chlordane, pp-DDE, pp-
DDD, pp-DDT, dieldrin, endrin, heptachlor,
heptachlor epoxide, lindane, methoxychlor),
and polychlorinated biphenyl (PCB; expressed
as Arochlor 1260) by gas chromatography with
electron capture detection (Agilent GC model
6890, Agilent Technologies, Palo Alto, Cali-
fornia, USA). The limits of quantification
(LOQ) for all OCs were 20 parts per billion
(ppb) with the exception of methoxychlor,
which had an LOQ of 250 ppb.
Whole blood, collected in buffered citrate
sodium tubes, was analyzed for detectable
arsenic, lead, and mercury at the University of
Pennsylvania. Arsenic and mercury were
determined by atomic absorption spectroscopy
(AAS) using hydride generation (AAnalyst 800
AA, Perkin Elmer, Wellesley, Massachusetts,
USA). The LOQs were 25 ppb for mercury
and 100 ppb for arsenic. Lead was determined
by graphite furnace AAS (AAnalyst 800,
Perkin-Elmer). The lead LOQ was 50 ppb.
All toxicant results were expressed on a wet
weight basis.
The distribution of each physical measure-
ment, hematologic, plasma biochemistry, and
plasma electrophoresis variable was evaluated
separately for the entire sample population,
age, gender, and behavior. The mean, stan-
dard deviation, median, and range were
determined for each parameter. The Shapiro-
Wilk statistic, kurtosis, and skewness were
used to evaluate the distribution of the data.
The X6SD are reported for normally distrib-
uted data; the median, 10%, and 90% quartiles
are reported for nonnormally distributed data.
Levene?s test for equality of variances was
used to determine if the data were homoge-
neous. Comparisons were made between age
groups, gender, and behavior. A one-way
analysis of variance (ANOVA) was used to
assess between group differences for normally
distributed data. Specific between-group dif-
ferences were evaluated using a Tukey?s test.
For data that were not normally distributed, a
Kruskal-Wallis one-way ANOVA and Dunn?s
test were used to assess differences between
and within groups, respectively. After comple-
tion of the crude analysis, a univariate general
linear model was used to evaluate behavior
and gender while controlling for age. Non-
normally distributed data were log trans-
formed for this analysis. A 432 exact test
was done to determine if physical exam
findings significantly differed between the
nesting, foraging, and stranded turtles. When
a difference was found at this level, a Fisher
exact test was used to further classify differ-
ences between groups. Historically, agreement
between different methods of measuring the
same biologic parameter has been determined
on the basis of correlation; however, this is not
considered optimal for agreement analysis
(Bland and Altman, 1999). In this study,
agreement between the two techniques for
evaluating the WBC counts was determined by
the Bland-Altman method (Bland and Altman,
1986, 1999). For this analysis, bias was defined
as the mean difference between the two
methods and limits of agreement were calcu-
lated as the bias 61.96 SD (Med-Calc
Software, Mariakerke, Belgium). Values of
P,0.05 were considered to reflect statistical
differences. Statistical analyses were per-
formed using SPSS 15.0 (SPSS Inc., Chicago,
Illinois, USA).
RESULTS
Physical examinations
Based on testosterone levels in the 39
foraging turtles, there were 30 females,
eight males, and one turtle of unknown
sex. Curved carapace length (mean6SD)
was 70.2 cm69.1 with a range of 55.6?
93.9 cm, and ages were determined as 35
subadults and four adults for the foraging
turtles. Thirty-one nesting adult females
received physical examinations and a CCL
of 99.6 cm65.8 and range of 87.0?
108.4 cm was recorded for 29 of these
females. Thirteen stranded turtles-seven
female, one male, five of unknown sex-
received physical examinations and were
classified as subadult (n59), adult (n53),
and unknown (n51). Curved carapace
length in 12 of these stranded turtles was
72.5 cm614.2 with a range of 56.0?
100.2 cm. Body weights were recorded
for all 39 foraging turtles (44.7 kg617;
range 20?105 kg) and 12 stranded turtles
(45.8 kg627.5; range 21.2?91.0 kg).
A number of clinical abnormalities were
detected in the turtles in each of the three
44 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
(39 foraging, 31 nesting, 13 stranded)
behaviors (Fig 1). There was a significant
difference in the number of heavy epibonts
on stranded turtles (62%) compared with
both nesting (6%, P50.0003) and foraging
turtles (29%, P50.03). There was also a
significant difference in the number of
epibonts on the foraging turtles compared
to the nesting turtles (P50.02). Both
stranded and nesting turtles were signifi-
cantly more likely to have miscellaneous
abnormalities (stranded: 38%, P50.002;
nesting: 19%, P50.03) compared with
foraging turtles (3%). There were signifi-
cant differences in the number of emaciat-
ed turtles in the stranded group (46%)
compared with both the nesting (0%)
(P50.0002) and foraging turtles (3%;
P50.0005), and weak turtles in the strand-
ed group (31%) compared with other
behaviors (0%, P,0.010). There was no
difference in the number of flipper ampu-
tations between the three groups (P50.4).
In the miscellaneous abnormalities, one
stranded turtle had a fish hook lodged in its
oral cavity; one had severe neurologic
clinical signs from a suspected spirorchid
trematode infestation based on clinical
signs, response to treatment, and similar
conditions affecting loggerheads in Florida
during this time period (Jacobson et al.,
2006); and one had a spindle cell sarcoma
within the cloacal region diagnosed by
surgical excision and histopathologic exam-
ination after 6 mo in rehabilitation post-
stranding. No fibropapillomatosis-like tu-
mors were observed on any of the 83
loggerheads examined.
Of the 10 stranded turtles evaluated for
epibiont loads, one had mild, one had
moderate, and eight had heavy infesta-
tions. Epibiont loads in foraging turtles
included seven mild, 15 moderate, and 10
heavy infestations, with seven loads not
recorded; nesting turtles had 13 mild, 11
moderate, and two heavy infestations, with
five loads not recorded.
Necropsies
Of the six stranded turtles that were
euthanized or died, necropsies revealed a
number of pathologic findings including
two turtles with heavy external epibiont
loads; two turtles with severe spirorchid
trematode infestations; two turtles with
significant carapace defects, one with
associated pneumonia and the other with
many barnacles observed on the coelomic
lining and thought to have entered via the
carapace defect; one turtle with hepatitis;
FIGURE 1. Clinical lesions noted in foraging, nesting, and stranded loggerhead sea turtles (Caretta caretta)
along the coast of Georgia.
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 45
and one turtle with osteomyelitis associat-
ed with a flipper lesion.
Hematology
Results of hematologic tests are provid-
ed in Table 1. The PCV was statistically
lower (P,0.005) in the stranded (0.19 l/l,
0.09?0.35) turtles than in the foraging
(0.32 l/l60.05) or nesting (0.30 l/l60.04)
turtles. Total solids were statistically dif-
ferent (P,0.05) between turtles exhibiting
all three behaviors, with nesting (48610 g/
L) turtles having the highest values and
stranded (2068 g/l) turtles the lowest
values. The foraging turtles had statistical-
ly lower WBC counts (9,00563,536) than
the nesting (P50.05) and stranded (P5
0.009) turtles using the eosinophil Unop-
ette method, but not by WBC estimate
using blood smears. Stranded turtles had
statistically higher (P,0.005) lymphocyte
counts (4,380, 912?11,700) and lower
monocyte counts (0, 0?150) than foraging
(P,0.005) and nesting (P50.003) turtles.
No hemoparasites were detected in any of
the 68 turtles tested. There was a low
degree of agreement between the two
WBC count measures (bias: 21018.9,
limits of agreement: 210282.7433 to
8244.8123; Fig. 2). Because of the poten-
tial for disagreement, the sampling tech-
niques were evaluated individually.
Plasma biochemistry
Plasma biochemistry values are provid-
ed in Table 2. Foraging turtles had higher
glucose (5.88 mmol/l61.11) than the
stranded (P50.002) and nesting (P5
0.001) turtles and higher potassium
(5.1 mmol/l62.0) values than turtles ex-
hibiting the other two behaviors (P,
0.001). Carbon dioxide values were statis-
tically higher (P,0.005) in stranded
(31 mmol/l, 28?36.5) versus foraging
(15 mmol/l6 4) and nesting (14.7 mmol/
l68.6) turtles. Nesting females had higher
cholesterol (6.94 mmol/l, 4.35?8.60) and
triglyceride values (4.54 mmol/l, 1.71?
9.75) and lower ALT values (4 U/l, 3?
127) than turtles exhibiting the other two
behaviors (P,0.005). Stranded turtles had
higher CK (1,366 U/l, 136?2,223) and UA
(77.32 mmol/l, 47.59?279.56) values than
turtles exhibiting the other two behaviors
(P,0.005). Significant differences in BUN
were found between turtles exhibiting the
various behaviors (P,0.005) with the
lowest values in nesting turtles (2.86
mmol/l, 2.00?4.43) and the highest in
foraging turtles (29.63 mmol/l, 14.28?
42.48). Total protein was statistically
different (P,0.005) between turtles ex-
hibiting various behaviors, with lowest
values (25 g/l69) in stranded turtles and
highest (52 g/l65) in nesting females.
Albumin values were statistically different
(P,0.005) between turtles exhibiting var-
ious behaviors, with lowest values (7 g/
l63) in stranded turtles and highest (17 g/
l62) in nesting females. Difference in
globulin values were also present with
values from foraging turtles (29 g/l69)
different from nesting (40 g/l67; P50.02)
and stranded (17 g/l67; P50.003) turtles;
nesting and stranded turtles were also
statistically different at P,0.005.
Plasma protein electrophoresis
Plasma protein electrophoresis values
are provided in Table 3. Nesting females
had significantly higher albumin values
(11.5 g/l62.9) than turtles exhibiting the
other two behaviors. There was no beta-
gamma bridging in any of the 71 turtles
evaluated.
Toxicants
Plasma OCs and PCBs were below their
LOQs in the 39 foraging, 11 nesting, and
three stranded turtles tested. There was
no statistical difference in arsenic levels
between the 23 foraging (7.91762.682
ppb), 12 nesting (1.82361.313 ppb), and
three stranded (3.83362.182 ppb) turtles
or for the lead levels between these same
23 foraging (0.08560.167 ppb), 12 nesting
(0.0966 0.041 ppb), and three stranded
(0.05060.000 ppb) turtles. There was a
significant difference (P,0.005) in the
46 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
mercury levels between the different
groups of turtles, with the nesting turtles
having higher mercury levels (0.3 ppb,
0.25?0.8) than foraging turtles (0.25 ppb,
0.25?0.40). There was no difference in the
mercury levels between the stranded
turtles (0.25 ppb, 0.25?10.0) and turtles
exhibiting either of the other two behav-
iors. There was a single stranded turtle
with an exponentially higher mercury level
(10 ppm) than that found in any other
turtle.
DISCUSSION
In this study a number of differences in
health parameters between foraging, nest-
ing, and stranded loggerhead turtles along
the coast of Georgia were noted. The
overall health of the foraging and nesting
turtles was rated as good based on physical
exam findings and blood values obtained,
although significant differences were not-
ed between turtles exhibiting the various
behaviors for many of these parameters. A
number of clinically significant physical
abnormalities were noted in the stranded
turtles, as well as significant differences in
blood parameters between the stranded
turtles and those exhibiting the other two
behaviors. However, because of study
constraints a different laboratory was used
to determine CBC and plasma biochem-
istry values in the stranded turtles. Other
limitations of this study were the nature in
which samples were collected (e.g., on a
boat and beach, and at a rehabilitation
facility) resulting in not all tests being
performed on all turtles because of poor
sample quantity and/or quality (e.g., he-
molysis) for some of these turtles. Blood
samples were processed within 4 hr (range
15 min?4 hr) for turtles exhibiting all the
behaviors and therefore any potential
influence of time on blood parameters
(e.g., glucose and potassium) would be
similar across behaviors.
TheWBCforforagingturtleswas
statistically lower than for the nesting
and stranded turtles using the eosinophil
Unopette method, but not based on
laboratory blood smear estimation. In this
study, values determined by the eosinophil
Unopette method and the blood smear
evaluations were statistically significantly
different for the stranded turtles (data not
shown) and may account for the differ-
ences between turtles exhibiting different
behaviors using the eosinophil Unopette
method, but not the blood smear estima-
tions.
Significant differences in the differen-
tial count between turtles exhibiting dif-
ferent behaviors included the increased
lymphocyte count in the stranded turtles,
which may indicate antigenic stimulation
of turtles within this behavior. Stranded
turtles also had lower monocyte counts,
but the significance of this finding is
unknown.
The low mean PCV for the stranded
turtles supports that anemia was present
in most of these turtles and likely associ-
ated with a chronic condition such as poor
nutrition, a chronic infectious or parasitic
disease, an immune deficiency related to
their debilitated state, or a combination of
some or all of these causes. Additionally,
the significantly lower total protein levels
in these stranded turtles also supports a
debilitated state associated with malnutri-
tion, parasitism, a protein-losing disease,
or a multifactorial condition. In contrast,
the nesting females had high protein
levels, likely a reflection of their egg laying
activity as seen in a number of reptile
species (Harr et al., 2001; Campbell,
2004).
Although epibiont load in this study was
determined on an ordinal scale, we did
observe a significant difference between
the behaviors, with .50% of the stranded
turtles having a heavy epibiont load on
their carapace; 26% of foraging turtles and
only 7% of nesting turtles had heavy
epibiont loads. The stranded turtles also
had significantly lower PCV and total
proteins as compared to the turtles
exhibiting other behaviors although we
can not make a direct association of these
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 47
T
ABLE
1.
Hematologic
values
in
foraging,
nesting,
and
stranded
loggerhead
sea
t
urtles
(
Caretta
caretta
)
along
the
coast
of
Georgia.
Parameter
(SI
units)
Foraging
a
Nesting
a
Stranded
b
Packed
cell
volume
(l/l)
(Mean
6
SD)
or
(Median,
10?90
%
quartiles)
0.32
6
0.05
0.30
6
0.04
0.19
c
,
0.09?0.35
Range
0.18?0.40
0.23?0.40
0.09?0.36
(
n
)
(39)
(34)
(11)
Total
solids
(g/l)
(Mean
6
SD)
37
6
12
c
48
6
10
c
20
6
8
c
Range
10?60
10?66
06?29
(
n
)
(39)
(33)
(6)
Red
blood
cells
(
3
10
3
/
m
l)
(Median,
10?90
%
quartiles)
520,000,
300,000?820,000
450,000,
298,000?704,000
130,000
Range
220,000?1,220,000
250,000?1,110,000
(
n
)
(39)
(30)
(1)
White
blood
cells
(
3
10
3
/
m
l)
d
(Mean
6
SD)
or
(median,
10?90
%
quartiles)
9,005
6
3,536
c
10,050,
6,690?18,620
17,900,
7,650?17,900
Range
4,000?17,400
4,000?23,950
7,650?24,450
(
n
)
(39)
(33)
(3)
White
blood
cells
(
3
10
3
/
m
l)
e
(Mean
6
SD)
9,022
6
2,081
9,658
6
2,881
11,042
6
4,505
Range
5,000?12,500
5,000?14,500
1,000?19,000
(
n
)
(39)
(18)
(12)
Heterophils
(
3
10
3
/
m
l)
(Mean
6
SD)
3,677
6
1,365
6,629
6
2,830
4,766
6
3,795
Range
345?7,164
2,400?14,220
200?14,060
(
n
)
(39)
(18)
(11)
Lymphocytes
(
3
10
3
/
m
l)
(Mean
6
SD)
or
(median,
10?90
%
quartiles)
2,725
6
1,150
2,298
6
732
4,380,
c
912?11,700
Range
299?4,830
900?3,300
780?33,430
(
n
)
(39)
(18)
(11)
Monocytes
(
3
10
3
/
m
l)
(Median,
10?90
%
quartiles)
960,
224?1,840
615,
261?1,860
0,
c
0?150
Range
64?2,750
0?1,950
0?150
(
n
)
(39)
(18)
(11)
48 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
Parameter
(SI
units)
Foraging
a
Nesting
a
Stranded
b
Eosinophils
(
3
10
3
/
m
l)
(Median,
10?90
%
quartiles)
1,152,
448?2,100
973,
298?2,521
0,
0?360
Range
192?3,584
105?2,530
0?380
(
n
)
(39)
(18)
(11)
Basophils
(
3
10
3
/
m
l)
(Median,
10?90
%
quartiles)
0
0,
0?18
0,
0?18
Range
0?180
6?70
(
n
)
(39)
(18)
(11)
Azurophils
(
3
10
3
/
m
l)
(Median,
10?90
%
quartiles)
0
0,
0?18
0,
0?523
Range
0?180
0?570
(
n
)
(39)
(18)
(11)
a
Values
obtained
at
the
University
of
Florida
Clinical
Pathology
Laborato
ry.
b
Values
obtained
at
Antech
Diagnostic
Laboratories,
Memphis,
Tennessee,
USA.
c
Significant
difference,
P
,
0.05.
d
White
blood
cell
count
performed
in
field
using
eosinophil
Unopette
syste
m.
e
White
blood
cell
count
performed
in
laboratory
by
absolute
count.
T
ABLE
1.
Continued.
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 49
blood parameters and epibiont loads
based on our data. This finding differs
from a study by Stamper et al. (2005) in
which no correlation was noted between
epibiont load and any blood parameters
evaluated in loggerhead turtles. However,
the turtles in that study were all captured
free-ranging in water and thus not debil-
itated and live-stranded as were turtles in
the stranded behavior in the present
study.
Foraging turtles had the highest glu-
cose, which is expected based on their
foraging state as compared to the fasting
state of most nesting females and the
debilitated condition of stranded turtles.
Additionally, higher potassium values
were found in the foraging turtles, which
may be due to decreased food intake in
nesting and debilitated turtles and possi-
ble physical abnormalities (e.g., diarrhea)
in stranded turtles.
Nesting turtles had significantly higher
cholesterol and triglyceride values than
turtles exhibiting the other two behaviors,
a finding compatible with vitellogenesis
and their egg-laying condition (Hamann et
al., 2003). Values from this study are
similar to values in nesting leatherback
turtles, cholesterol (7.58 mmol/l6 1.89)
and triglyceride (4.65 mmol/l6 0.40;
Deem et al., 2006), but differs from
previously published cholesterol (2.74
mmol/l) and triglyceride (0.98 mmol/l)
values for free-ranging foraging logger-
head juvenile and adult turtles (Bolten et
al., 1992), values similar to the foraging
loggerheads in this study. The higher total
protein, albumin, and globulin values for
nesting females is also compatible with the
reproductive state of turtles in this behav-
ior (Harr et al., 2001; Campbell, 2004).
Bloodureanitrogenvaluesdifered
among turtles exhibiting the three behav-
iors with nesting turtles having the lowest
values; most likely a reflection of their
fasting state. The statistically higher UA of
the stranded turtles may be associated
with the debilitated state of these turtles
and dehydration. The CK value was
FIGURE 2. A Bland-Altman plot measuring the level of agreement between two different methods for
evaluating white blood cell counts in loggerhead sea turtles (Caretta caretta) along the coast of Georgia.
50 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
T
ABLE
2.
Plasma
biochemistry
values
in
foraging,
nesting,
and
stranded
loggerh
ead
sea
turtles
(
Caretta
caretta
)
along
the
coast
of
Georgia.
Parameter
(SI
units)
Foraging
a
Nesting
a
Stranded
b
Glucose
(mmol/l)
(Mean
6
SD)
5.88
6
1.11
c
5.33
6
0.83
4.50
6
2.1
Range
3.9?7.6
4.1?6.3
1.72?7.66
(
n
)
(38)
(25)
(13)
Sodium
(mmol/l)
(Mean
6
SD)
156
6
11
148
6
6
155
6
8
Range
135?175
139?162
146?174
(
n
)
(38)
(25)
(13)
Potassium
(mmol/l)
(Mean
6
SD)
5.1
6
2.0
c
4.0
6
1.0
3.7
6
0.3
Range
3.3?13.9
3?5
3.3?4.4
(
n
)
(38)
(25)
(13)
Chloride
(mmol/l)
(Mean
6
SD)
130
6
11
115
6
4
121
6
6
Range
107?158
110?123
113?134
(
n
)
(16)
(8)
(9)
CO
2
(mmol/l)
(Mean
6
SD)
or
(median,
10?90
%
quartiles)
15
6
4
14.7
6
8.6
31,
c
28?36.5
Range
10?24
5.0?34.0
25?38
(
n
)
(39)
(25)
(5)
Blood
Urea
Nitrogen
(mmol/l)
(Median,
10?90
%
quartiles)
29.63,
c
14.28?42.48
2.86,
c
2.00?4.43
24.63,
c
9.35?33.24
Range
0.357?38.2
1.79?4.64
9.28?33.92
(
n
)
(39)
(25)
(12)
Creatinine
(
m
mol/l)
(Median,
10?90
%
quartiles)
26.52,
17.68?53.04
8.84,
8.84?30.06
35.36,
8.84?57.46
Range
8.84?44.2
8.84?53.04
8.84?70.72
(
n
)
(39)
(25)
(6)
Total
protein
(g/l)
(Mean
6
SD)
37
6
11
c
52
6
5
c
25
6
9
c
Range
16?56
46?61
4?39
(
n
)
(39)
(25)
(13)
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 51
Parameter
(SI
units)
Foraging
a
Nesting
a
Stranded
b
Albumin
(g/l)
(Mean
6
SD)
13
6
3
c
17
6
2
c
7
6
3
c
Range
8?16
14?19
3?12
(
n
)
(12)
(8)
(13)
Globulin
(g/l)
(Mean
6
SD)
29
6
9
c
40
6
7
c
17
6
7
c
Range
10?40
27?51
1?24
(
n
)
(12)
(8)
(13)
Cholesterol
(mmol/l)
(Median,
10?90
%
quartiles)
1.94,
1.17?4.51
6.94,
c
4.35?8.60
2.03,
0.13?2.25
Range
1.17?5.18
4.90?8.78
0.13?5.05
(
n
)
(39)
(25)
(9)
Triglyceride
(mmol/l)
(Median,
10?90
%
quartiles)
0.62,
0.25?1.38
4.54,
c
1.71?9.75
0.11,
0.11?0.25
Range
0.17?1.38
4.41?4.89
0.11?0.29
(
n
)
(39)
(25)
(5)
Calcium
(mmol/l)
(Mean
6
SD)
or
(Median,
10?90
%
quartiles)
1.85,
1.48?2.35
2.03
6
1.05
1.45
6
0.25
Range
1.4?2.08
0.65?3.18
1.08?1.9
(
n
)
(39)
(25)
(13)
Phosphorus
(mmol/l)
(Mean
6
SD)
2.07
6
0.36
2.23
6
0.58
2.33
6
0.39
Range
1.32?2.55
2.03?3.59
1.74?3.10
(
n
)
(39)
(25)
(13)
Uric
acid
(
m
mol/l)
(Median,
10?90
%
quartiles)
41.64,
23.80?89.22
23.79,
11.90?47.58
77.32,
c
47.58?279.56
Range
11.90?71.38
11.90?53.53
11.09?297.4
(
n
)
(39)
(25)
(13)
Alanine
aminotransferase
(U/l)
(Mean
6
SD)
or
(median,
10?90
%
quartiles)
16
6
6
4.0,
c
3?127
22
6
13
Range
0?29
3?30
10?44
(
n
)
(39)
(25)
(6)
T
ABLE
2.
Continued.
52 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
Parameter
(SI
units)
Foraging
a
Nesting
a
Stranded
b
Aspartate
aminotransferase
(U/l)
(Median,
10?90
%
quartiles)
165,
118?256
157,
129?260
199,
138?941
Range
2?255
116?190
113?1,199
(
n
)
(39)
(25)
(13)
Lactate
dehydrogenase
(U/l)
(Median,
10?90
%
quartiles)
572,
229?1,401
592,
255?1,141
429,
244?3,057
Range
6?1,376
22?1,172
244?3,876
(
n
)
(39)
(25)
(5)
Creatine
kinase
(U/l)
(Median,
10?90
%
quartiles)
534,
243?1,075
359,
147?1,392
1,366,
c
136?2,223
Range
3?1,899
81?1,627
100?26,070
(
n
)
(39)
(25)
(13)
Amylase
(U/l)
(Mean
6
SD)
263
6
99
407
6
131
180
6
106
Range
2?417
176?593
50?307
(
n
)
(39)
(25)
(5)
Lipase
(U/l)
(Median,
10?90
%
quartiles)
1.0,
1?9
20.0,
1?42
5.5,
1?50
Range
1?14
13?49
1?63
(
n
)
(39)
(25)
(5)
Gamma
glutamyl
transferase
(U/l)
(Median,
10?90
%
quartiles)
9.0,
5?10
8.0,
5?10
7.5,
6?9
Range
5?15
5?14
6?9
(
n
)
(39)
(25)
(5)
a
Values
obtained
at
the
University
of
Miami,
Department
of
Pathology.
b
Values
obtained
at
Antech
Diagnostic
Laboratories,
Memphis,
Tennessee,
USA.
c
Significant
difference,
P
,
0.05.
T
ABL
E
2.
Continued.
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 53
significantly higher in the stranded turtles
as compared to the other turtles, consis-
tent with muscle injury and wasting noted
in a number of these debilitated turtles.
Additionally, the higher CO
2
value may be
a reflection of decreased respiratory ven-
tilation in the stranded, debilitated turtles.
The only significant difference in plas-
ma protein electrophoresis between the
turtles was a higher albumin value for
nesting females. There was no beta-
gamma bridging in any of the loggerhead
turtles in our study, which differs from a
study in which bridging was found in 28%
of 41 clinically normal loggerhead sea
turtles tested from Florida waters (Gicking
et al., 2004). Beta-gamma bridging is often
associated with chronic disease or para-
sites (Gicking et al., 2004) so it is
interesting to note the absence of this
finding in all of the turtles in this study,
especially those in the stranded behavior.
Additionally, we noted a discrepan-
cy within behaviors in protein values
obtained using the three methods-refrac-
tometer and chemistry analyzer (total
proteins), and chemisty analyzer and
SPEP-II agarose gels (albumin)-which is
consistent with findings for other species
(Lumeij et al., 1990). This discrepancy
should be taken into consideration when
one is comparing the results between
studies.
A number of studies have examined
loggerhead turtles for PCBs, OC insecti-
cides, and metals (Gordon et al., 1998;
Sakaietal.,2000;Dayetal.,2005;
Gardner et al., 2003; Franzellitti et al.,
2004; Keller et al., 2004a, b; Storelli et al.,
2005). These studies have measured either
PCBs and OCs or metals, but not both.
Additionally, most studies have involved
assessment of tissues from dead animals.
Thus, there are a limited number of
studies in which direct comparisons to
the data from this study can be made.
TABLE 3. Plasma protein fractions identified in foraging, nesting, and stranded loggerhead sea turtles
(Caretta caretta) along the coast of Georgia, USA.
Parameter (SI units) Foraging Nesting Stranded
Pre-albumin (g/L)
(Median, 10?90% quartiles) 0.0, 0?0.1 0.8, 0?0.11 0
Range 0.00?1.0 0.00?1.2
(n) (39) (24) (8)
Albumin (g/L)
(Mean 6 SD) 7.962.6 11.562.9
a
6.363.7
Range 3.1?12.4 10.4?18.3 2.3?12.0
(n) (39) (24) (8)
Alpha-1 (g/L)
(Mean 6 SD) 1.460.6 1.560.6 0.960.3
Range 0.5?2.5 0.5?2.7 0.3?1.3
(n) (39) (24) (8)
Alpha-2 (g/L)
(Median, 10?90% quartiles) 1.2, 0.5?2.5 2.3, 1.5?6.5 1.5, 0.9?2.7
Range 0.5?3.1 1.1?10.0 0.9?3.3
(n) (39) (24) (8)
Beta (g/L)
(Mean 6 SD) 9.962.8 17.165.2 7.464.5
Range 4.5?14.6 8.7?25.9 2.2?15.8
(n) (39) (24) (8)
Gamma (g/L)
(Mean 6 SD) 15.767.4 11.863.7 9.563.6
Range 5.1?28.8 7.4?21.6 4.7?15.7
(n) (39) (24) (8)
a
Significant difference P,0.05.
54 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009
There is little information regarding
whole blood mercury, lead, and arsenic
concentrations in sea turtles generally and
loggerhead turtles specifically. The mean
whole blood lead and total mercury
concentrations from 106 Kemp?s ridley
sea turtles were 11 ppb and 18 ppb,
respectively (Kenyon et al., 2001). The
mean whole blood mercury concentrations
for34livecapturedand6stranded
loggerhead turtles was 29 ppb and
99 ppb, respectively (Day et al., 2005).
The detected concentrations in the cur-
rent study are below these levels, with
nesting turtles having higher mercury
concentrations than turtles exhibiting the
other two behaviors.
The mean concentration of total PCBs
in the plasma of 12 juvenile loggerhead
sea turtles off the coast of North Carolina
was 7.13 ppb (64.94 ppb), below the
LOQ in our study (Keller et al., 2004a,
b). Although such concentrations are low,
a variety of adverse health effects have
been postulated (Keller et al., 2004a, b)
and thus a lower LOQ than that used in
the current study is recommended for
future studies.
Although a few limitations exist with the
present study, we provide baseline blood
values for two cohorts (foraging and
nesting) of loggerhead sea turtles on the
coast of Georgia. Additionally, we demon-
strate significant differences in clinical
findings and blood parameters between
foraging, nesting, and stranded loggerhead
turtles in the region.
ACKNOWLEDGMENTS
We gratefully acknowledge V. Greco for her
laboratory technical support. Research for this
project was performed under Georgia Depart-
ment of Natural Resources permit (29-WMB-
01-140). Funding and logistic support for this
project was provided by the Lawrence Foun-
dation, the Smithsonian National Zoological
Park, St. Catherines Island Foundation, Wild-
life Conservation Society, the Georgia and
South Carolina Departments of Natural Re-
sources, the US Fish and Wildlife Service on
Blackbeard Island, Georgia, and Georgia
Southern University.
LITERATURE CITED
BLAND, J. M., AND D. G. ALTMAN. 1986. Statistical
methods for assessing agreement between two
methods of clinical measurement. Lancet 1:
307?310.
???, AND ???. 1999. Measuring agreement in
method comparison studies. Statistical Methods
in Medical Research 8: 135?160.
BOLTEN, A. B., E. R. JACOBSON, AND K. A. BJORNDAL.
1992. Effects of anticoagulant and autoanalyzer
on blood biochemical values of loggerhead sea
turtles (Caretta caretta). American Journal of
Veterinary Research 53: 2224?2227.
CAMPBELL, T. W. 2004. Clinical chemistry of reptiles.
In Veterinary hematology and clinical chemistry,
M. A. Thrall (ed.). Lippincott Williams and
Wilkins, Philadelphia, Pennsylvania, pp. 497.
CRAY, C., AND L. M. TATUM. 1998. Applications of
protein electrophoresis in avian diagnostics.
Journal of Avian Medicine and Surgery 12: 4?10.
DAY, R. D., S. J. CHRISTOPHER,P.R.BECKER, AND D.
W. WHITAKER. 2005. Monitoring mercury in the
loggerhead sea turtle, Caretta caretta. Environ-
mental Science & Technology 39: 437?446.
DEEM, S. L., E. S. DIERENFELD,G.P.SOUNGUET,A.
R. ALLEMAN,C.CRAY,R.H.POPPENGA,T.M.
NORTON, AND W. B. KARESH. 2006. Blood values
in free-ranging nesting leatherback sea turtles
(Dermochelys coriacea) on the coast of the
Republic of Gabon. Journal of Zoo and Wildlife
Medicine 37: 464?471.
FRANZELLITTI, S., C. LOCATELLI,G.GEROSA,C.
VALLINI, AND E. FABBRI. 2004. Heavy metals in
tissues of loggerhead turtles (Caretta caretta)
from the northwestern Adriatic Sea. Compara-
tive Biochemistry and Physiology 138: 187?194.
GARDNER, S. C., M. D. PIER,R.WESSELMAN, AND J. A.
JUAREZ. 2003. Organochlorine contaminants in
sea turtles from the Eastern Pacific. Marine
Pollution Bulletin 46: 1082?1089.
GEORGE, R. H. 1997. Health problems and diseases
of sea turtles. In The biology of sea turtles, P. L.
Lutz and J. A. Musick (eds.). CRC Press, Boca
Raton, Florida, pp. 363?385.
GICKING, J. C., A. M. FOLEY,K.E.HARR,R.E.
RASKIN, AND E. JACOBSON. 2004. Plasma protein
electrophoresis of the Atlantic loggerhead sea
turtle (Caretta caretta). Journal of Herpetolog-
ical Medicine and Surgery 14: 13?18.
GORDON, A. N., A. R. POPLE, AND J. NG. 1998. Trace
metal concentrations in livers and kidneys of sea
turtles from south-eastern Queensland, Austra-
lia. Marine and Freshwater Research 49: 409?
414.
HAMANN, M., C. J. LIMPUS, AND D. W. OWENS. 2003.
Reproductive cycles of males and females. In
The biology of sea turtles: Volume II, P. L. Lutz,
J. A. Musick and J. Wyneken (eds.). CRC Press,
Washington, D.C, pp. 135?161.
DEEM ET AL.?BLOOD VALUES IN LOGGERHEAD TURTLES 55
HARR, K. E., A. R. ALLEMAN,P.M.DENNIS,L.K.
MAXWELL,B.A.LOCK,R.A.BENNETT, AND E.
JACOBSON. 2001. Morphologic and cytochemical
characteristics of blood cells and hematologic
and plasma biochemical reference ranges in
green iguanas. Journal of the American Veteri-
nary Medical Association 218: 915?921.
HARVEY, J. W. 2001. Examination of blood samples.
In Atlas of veterinary hematology, J. W. Harvey
(ed.). WB Saunders, Philadelphia, Pennsylvania,
pp. 3?20.
HAWKEY, C. M., AND T. B. DENNETT. 1989. Normal
and abnormal red cells, granulocytes, lympho-
cytes, monocytes and azurophils. In Color atlas
of comparative veterinary hematology, C. M.
Hawkey and T. B. Dennett (eds.). Iowa State
University Press, Ames, Iowa, pp. 58?138.
HERBST, L. H., AND E. R. JACOBSON. 2003. Practical
approaches for studying sea turtle health and
disease. In The biology of sea turtles: Volume II,
P. L. Lutz, J. A. Musick and J. Wyneken (eds.).
CRC Press, Washington, D.C., pp. 385?410.
JACOBSON, E. R., B. L. HOMER,B.A.STACY,E.C.
GREINER,N.J.SZABO,C.L.CHRISMAN,F.ORIGGI,
S. COBERLEY,A.M.FOLEY,J.H.LANDSBERG,L.
FLEWELLING,R.Y.EWING,R.MORETTI,S.SCHAF,
C. ROSE,D.R.MADER,G.R.HARMAN,C.A.
MANIRE,N.S.METTEE,A.P.MIZISIN, AND G. D.
SHELTON. 2006. Neurological disease in wild
loggerhead sea turtles (Caretta caretta). Diseas-
es of Aquatic Organisms 70: 139?154.
KELLER, J. M., J. R. KUCKLICK,M.A.STAMPER,C.A.
HARMS, AND P. D. MCCLELLAN-GREEN. 2004a.
Associations between organochlorine contami-
nant concentrations and clinical health parame-
ters in loggerhead sea turtles from North
Carolina, USA. Environmental Health Perspec-
tives 112: 1074?1079.
???, P. D. MCCLELLAN-GREEN,J.R.KUCKLICK,D.
E. KEIL, AND M. M PEDEN-ADAMS. 2004b. Effects
of organochlorine contaminants on loggerhead
sea turtle immunity: Comparison of a correlative
field study and in vitro exposure experiments.
Environmental Health Perspectives 114: 70?76.
KENYON, L. O., A. M. LANDRY, AND G. A. GILL. 2001.
Trace metal concentrations in blood of Kemp?s
ridley sea turtles (Lepidochelys kempii). Chelo-
nian Conservation and Biology 4: 129?135.
LUMEIJ, J. T., J. J. DEBRUIJE, AND M. M. KWANT. 1990.
Comparison of different methods of measuring
protein and albumin in pigeon sera. Avian
Pathology 19: 255?261.
MAIER, P. P., A. L. SEGARS,M.D.ARENDT,J.D.
WHITAKER,B.W.STENDER,L.PARKER,R.
VENDETTI,D.W.OWENS,J.QUATTRO, AND S. R.
MURPHY. 2004. Development of an index of sea
turtle abundance based upon in-water sampling
with trawl gear. Final project report to The
National Marine Fisheries Service. National
Oceanic and Atmospheric Administration,
Charleston, South Carolina, March 31, 2004.
Grant NA07FL0499.
NORTON, T. M. 2005. Sea turtle conservation in
Georgia and an overview of the Georgia sea
turtle center on Jekyll Island, Georgia. Georgia
Journal of Science 63: 208?230.
???, M. DODD,A.SEGARS,J.M.KELLER,M.
PEDEN-ADAMS,R.D.DAY,C.HARMS,E.JACOB-
SON,A.FOLEY,S.MURPHY,W.CLUSE,W.TEAS,
M. BRESETTE,B.SCHROEDER,A.MACKINNON, AND
N. STEDMAN. 2005. Debilitated loggerhead turtle
(Caretta caretta) syndrome along the southeast-
ern US coast: Incidence, pathogenesis, and
monitoring. In Proceedings of the twenty-fifth
annual symposium of sea turtle biology and
conservation. Sea turtle symposium 2005, NOAA
Technical Memorandum, Savannah, Georgia.
OWENS, D. W., AND G. J. RUIZ. 1980. New methods of
obtaining blood and cerebrospinal fluid from
marine turtles. Herpetologica 36: 17?20.
???, J. R. HENDRICKSON,V.LANCE, AND I. P.
CALLARD. 1978. A technique for determining sex
of immature Chelonia mydas using a radioim-
munoassay. Herpetologica 34: 270?273.
SAKAI, H., K. SAEKI,H.ICHIHASHI,H.SUGANUMA,S.
TANABE, AND R. TATSUKAWA. 2000. Species
specific distribution of heavy metals in tissues
and organs of loggerhead turtle (Caretta caretta)
and green turtle (Chelonia mydas) from Japa-
nese coastal waters. Marine Pollution Bulletin
40: 7801?709.
STAMPER, M. A., C. HARMS,S.P.EPPERLY,J.BRAUN-
MCNEILL,L.AVENS, AND M. K. STOSKOPF. 2005.
Relationship between barnacle epibiotic load
and hematologic parameters in loggerhead sea
turtles (Caretta caretta), a comparison between
migratory and residential animals in Pamlico
Sound, North Carolina. Journal of Zoo and
Wildlife Medicine 36: 635?641.
STORELLI, M. M., A. STORELLI, R. D?ADDABBO,C.
MARANO,R.BRUNO, AND G. O. MARCOTRIGIANO.
2005. Trace elements in loggerhead turtles
(Caretta caretta) from the eastern Mediterra-
nean Sea: Overview and evaluation. Environ-
mental Pollution 135: 163?170.
WHITAKER, B. R., AND H. KRUM. 1999. Medical
management of sea turtles in aquaria. In Zoo and
wild animal medicine: Current therapy 4, M. E.
Fowler and R. E. Miller (eds.). WB Saunders,
Philadelphia, Pennsylvania, pp. 217?231.
Received for publication 8 June 2007.
56 JOURNAL OF WILDLIFE DISEASES, VOL. 45, NO. 1, JANUARY 2009