Justin R. Perrault

Numerous toxin-producing harmful algal (HAB) species occur in Florida’s coastal waters. Exposure to these toxins has been shown to have sublethal effects in sea
turtles. The objective of this study was to establish concentrations of 10 HAB toxins in plasma samples from green turtles (Chelonia mydas) foraging in Florida’s Big
Bend. Domoic acid, lyngbyatoxin-A, microcystins, nodularin, and okadaic acid were detected, demonstrating exposure to these HAB toxins, which are also a public
health concern.

Important indicators of population health needed for large-scale sea turtle population
recovery efforts include demographics, disease and mortality trends, condition indices, and
baseline blood data. With this comprehensive health assessment of adult female green sea turtles
Chelonia mydas nesting on Juno Beach, Florida, USA, we (1) established baseline health indices;
(2) identified individuals with evidence of infection by chelonid alphaherpes viruses 5 and 6
(ChHV5, ChHV6), which are implicated in fibropapillomatosis and respiratory and skin disease,
respectively; and (3) compared measured health indices between turtles that did versus those that
did not test positive for ChHV5 and/or ChHV6. All 60 turtles included in the study were in good
body condition with no external fibropapillomatosis tumors. Hematological and biochemical reference
intervals were established. Via quantitative PCR (qPCR), 5/60 turtles (8%) tested positive
for ChHV5, and all turtles were negative for ChHV6. Of 41 turtles tested for antibodies to ChHV5
and ChHV6, 29% and 15% tested positive, respectively, and 10% tested positive for antibodies to
both viruses. Notably, there were no statistically significant differences between health variables
for nesting turtles that tested positive for ChHV5 DNA versus those that tested negative; and also
no differences between turtles that tested positive for ChHV5 or ChHV6 antibodies and those that
did not. This suggests that these viruses are enzootically stable in Florida’s adult green turtles.
This study provides a health profile of nesting green turtles in southeastern Florida applicable to
temporal and spatial investigations of this and other populations.

Anthropogenic contaminants in the marine environment often biodegrade slowly,
bioaccumulate in organisms, and can have deleterious effects on wildlife immunity,
health, reproduction, and development. In this study, we evaluated tissue toxicant
concentrations and pathology data from 83 odontocetes that stranded in the
southeastern United States during 2012–2018. Mass spectrometry was used to
analyze blubber samples for five organic toxicants (atrazine, bisphenol-A, diethyl
phthalates, nonylphenol monoethoxylate [NPE], triclosan), and liver samples were
analyzed for five non-essential elements (arsenic, cadmium, lead, mercury, thallium),
six essential elements (cobalt, copper, manganese, iron, selenium, zinc) and one
toxicant mixture class (Aroclor1268). Resultant data considerably improve upon the
existing knowledge base regarding toxicant concentrations in stranded odontocetes.
Toxicant and element concentrations varied based on animal demographic factors
including species, sex, age, and location. Samples from bottlenose dolphins had
significantly higher average concentrations of lead, manganese, mercury, selenium,
thallium, and zinc, and lower average concentrations of NPE, arsenic, cadmium, cobalt,
and iron than samples from pygmy sperm whales. In adult female bottlenose dolphins,
average arsenic concentrations were significantly higher and iron concentrations were
significantly lower than in adult males. Adult bottlenose dolphins had significantly higher average concentrations of lead, mercury, and selenium, and significantly lower average
manganese concentrations compared to juveniles. Dolphins that stranded in Florida had
significantly higher average concentrations of lead, mercury, and selenium, and lower
concentrations of iron than dolphins that stranded in North Carolina. Histopathological
data are presented for 72 animals, including microscopic evidence of Campula spp.
and Sarcocystis spp. infections, and results of Morbillivirus and Brucella spp. molecular
diagnostic testing. Sublethal cellular changes related to toxicant exposure in free-ranging
odontocetes may lead to health declines and, in combination with other factors, may
contribute to stranding.

Fibropapillomatosis is a debilitating tumor disease of sea turtles that is sometimes
fatal. This disease is a key concern for sea turtle rehabilitation facilities due to its infectious nature,
as it is associated with a virus called chelonid alphaherpesvirus 5. This is the first study to analyze
antibodies to this virus in loggerhead sea turtles and represents the most complete dataset on
viral detection in sea turtles encountered in the more northern latitudes of their habitat in the
western Atlantic.

Chelonid alphaherpesviruses 5 and 6 (ChHV5 and ChHV6) are viruses that affect wild
sea turtle populations. ChHV5 is associated with the neoplastic disease fibropapillomatosis (FP),
which affects green turtles (Chelonia mydas) in panzootic proportions. ChHV6 infection is associated
with lung-eye-trachea disease (LETD), which has only been observed in maricultured sea turtles,
although antibodies to ChHV6 have been detected in free-ranging turtles. To better understand
herpesvirus prevalence and host immunity in various green turtle foraging aggregations in Florida,
USA, our objectives were to compare measures of innate and adaptive immune function in relation
to (1) FP tumor presence and severity, and (2) ChHV5 and ChHV6 infection status. Free-ranging,
juvenile green turtles (N = 45) were captured and examined for external FP tumors in Florida’s Big
Bend, Indian River Lagoon, and LakeWorth Lagoon. Blood samples were collected upon capture
and analyzed for ChHV5 and ChHV6 DNA, antibodies to ChHV5 and ChHV6, in vitro lymphocyte
proliferation using a T-cell mitogen (concanavalin A), and natural killer cell activity. Despite an
overall high FP prevalence (56%), ChHV5 DNA was only observed in one individual, whereas 20% of
turtles tested positive for antibodies to ChHV5. ChHV6 DNA was not observed in any animals and
only one turtle tested positive for ChHV6 antibodies. T-cell proliferation was not significantly related
to FP presence, tumor burden, or ChHV5 seroprevalence; however, lymphocyte proliferation in
response to concanavalin A was decreased in turtles with severe FP (N = 3). Lastly, green turtles with
FP (N = 9) had significantly lower natural killer cell activity compared to FP-free turtles (N = 5). These
results increase our understanding of immune system effects related to FP and provide evidence that
immunosuppression occurs after the onset of FP disease.

The gopher tortoise (Gopherus polyphemus), a keystone species, is declining throughout its geographic range. Lack of
knowledge with respect to the potential infectious diseases present within wild populations creates a dilemma for wildlife
biologists, conservationists and public policy makers. The objective of this study was to conduct a health assessment of two
previously unstudied gopher tortoise aggregations located at two sites in southeastern FL. Samples were collected from
91 tortoises (48 adults, 35 juveniles, 8 hatchlings) captured at Florida Atlantic University’s Harbor Branch Oceanographic
Institute, in Fort Pierce, FL, USA in 2019, and Loggerhead Park in Juno Beach, FL, USA, during 2018–2019. Samples of blood,
nasal swabs and oral/cloacal swabs were analyzed for hematology, plasma protein electrophoretic profiles and infectious
disease testing including Mycoplasma spp. serology and polymerase chain reaction (PCR) assays for Ranavirus, Herpesvirus and
Anaplasma spp. Hematological and plasma protein electrophoresis reference intervals are presented for adult and juvenile
tortoises from both sites combined. Clinical signs consistent with upper respiratory tract disease (URTD) were observed in
18/91 (20%) tortoises, and antibodies to Mycoplasma agassizii were detected in 33/77 (42.9%) tortoises. Adult tortoises were
significantly more likely than juveniles to have URTD clinical signs, and statistically significant, positive relationships were
observed between the presence of antibodies to Mycoplasma spp. and carapace length, packed cell volume and plasma
globulin concentrations. Anaplasma spp. inclusions were observed in 8/82 (10%) tortoises, but PCR detected Anaplasma sp.
in 21/83 (25%) tortoises. Herpesvirus and Ranavirus were not detected in any blood or swab samples. This work contributes
important baseline information on the health of gopher tortoises toward the southern end of the species’ range.

Sea turtles face both anthropogenic and natural threats including boat strikes, fisheries,
pollution, and predator attacks. Injuries from anthropogenic sources are more common than
naturally caused injuries. The goal of this study was to determine prevalence and cause (e.g. boat
strike, entanglement, hook, shark bite) of injuries on nesting loggerhead sea turtles Caretta
caretta on Juno and Jupiter beaches, Florida, USA. During the 2019 and 2020 nesting seasons, 450
loggerhead females were examined for external injuries. Injuries were categorized by anatomic
location, condition, and cause. We found that 24% of loggerheads had at least 1 injury. Of the 111
injuries found on 107 nesting females, 88% were healed, 9% were partially healed with some
scarred tissue, and 3% were fresh injuries. Most injuries (55%) were lateral injuries on the carapace
or appendages. We were able to attribute 60 injuries to a specific cause. Boat strikes
accounted for 75% of the 60 injuries, shark bites accounted for 15%, fishing hooks accounted for
7%, and entanglements accounted for the remaining 3%. This study provides new insight into the
prevalence of anthropogenic injuries relative to natural injuries in loggerhead sea turtles nesting
in the most densely nested beach in the Western Hemisphere and can be used to improve conservation
management plans through implementation of fishing and/or boating restrictions in the
nesting and foraging areas most commonly frequented by sea turtles.

Fibropapillomatosis (FP), a debilitating, infectious neoplastic disease, is rarely reported
in endangered Kemp’s ridley sea turtles (Lepidochelys kempii). With this study, we describe FP and
the associated chelonid alphaherpesvirus 5 (ChHV5) in Kemp’s ridley turtles encountered in the
United States during 2006–2020. Analysis of 22 case reports of Kemp’s ridley turtles with FP revealed
that while the disease was mild in most cases, 54.5% were adult turtles, a reproductively valuable
age class whose survival is a priority for population recovery. Of 51 blood samples from tumor-free
turtles and 12 tumor samples from turtles with FP, 7.8% and 91.7%, respectively, tested positive for
ChHV5 DNA via quantitative polymerase chain reaction (qPCR). Viral genome shotgun sequencing
and phylogenetic analysis of six tumor samples show that ChHV5 sequences in Kemp’s ridley
turtles encountered in the Gulf of Mexico and northwestern Atlantic cluster with ChHV5 sequences
identified in green (Chelonia mydas) and loggerhead (Caretta caretta) sea turtles from Hawaii, the southwestern Atlantic Ocean, and the Caribbean. Results suggest an interspecific, spatiotemporal
spread of FP among Kemp’s ridley turtles in regions where the disease is enzootic. Although FP is
currently uncommon in this species, it remains a health concern due to its uncertain pathogenesis
and potential relationship with habitat degradation.

At the time of hatchling emergence from a nest laid on Juno Beach, Florida, US, by a
normally pigmented green turtle (Chelonia mydas), 23 albino hatchlings and 75 normally pigmented
hatchlings were observed. This condition is rarely seen in sea turtles, and little is known about blood
analytes and genetics of albino wildlife to date. Therefore, the objective of our study was to assess and
compare morphometric measurements (mass, minimum straight carapace length, body condition
index), carapacial scute anomalies, a suite of hematologic and plasma biochemical analytes, and two
glucose analysis methodologies (glucometer and dry chemistry analysis) in albino (n¼20) versus
normally pigmented (n¼24) hatchlings from this nest. Genetic analyses were completed to identify
paternal contributions of hatchlings and to test Mendelian inheritance assumptions. Although
morphometric measurements, scute anomalies, and leukocyte morphology were similar between
albino and normally pigmented hatchlings, several differences were observed in blood analyte data:
immature erythrocytes, packed cell volume, heterophil:lymphocyte ratio, and glucose concentrations
(by both methodologies) were significantly higher, whereas absolute immature heterophils, absolute
lymphocytes, number of erythrocyte micronuclei, sodium, and chloride were significantly lower in
albino hatchlings compared with normally pigmented hatchlings. Considerations for these differences
include a stress response from sampling (e.g., timing of procedures or possibly from photosensitivity or
reduced visual acuity in albinos) and different osmoregulation, which may reflect physiologic variations
or stress. There was a small positive bias (0.10 mmol/L) with glucose by glucometer, similar to reports in
other sea turtle species and confirming its suitability for use in hatchlings. All albino hatchlings analyzed
(n¼10) were from the same father, but the normally pigmented hatchlings (n¼24) were from two other
fathers. These findings provide insight into the physiology and genetics of albinism in sea turtles.
Key words: Biochemistry, genotype, glucose, hematology, leucism, method comparison, multiple
paternity, sea turtle.