Insights on Immune Function in Free-Ranging Green Sea Turtles (Chelonia mydas) with and without Fibropapillomatosis.

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 Lake Worth 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.

Plasma chemistry in nesting leatherback sea turtles (Dermochelys coriacea) from Florida: Understanding the importance of sample hemolysis effects on blood analytes.

Plasma chemistry is widely used in diagnostic and research settings in sea turtles. However, plasma discolorations such as hemolysis are often not considered in data interpretation. The objectives of this study were to (1) evaluate the effects of moderate hemolysis on plasma electrolytes, minerals, and proteins using dry chemistry analysis (DCA) and protein electrophoresis from nesting leatherback sea turtles (Dermochelys coriacea) from Florida and to (2) establish blood analyte reference intervals. Twenty-six plasma samples with absence of hemolysis were selected and sub-divided into one non-hemolytic aliquot and an aliquot that was experimentally manipulated to mimic moderate hemolysis. Plasma samples were analyzed for hemoglobin using a handheld photometer; sodium, potassium, chloride, magnesium, calcium, phosphorus, and total protein using DCA; and protein electrophoresis. Packed cell volume and hemoglobin were measured in corresponding whole blood samples. Reference intervals were established. All analytes except calcium and pre-albumin were significantly higher and the calcium:phosphorus and albumin:globulin ratios were significantly lower in hemolytic plasma compared to non-hemolytic plasma. Alpha2-globulins and potassium were the analytes most impacted by hemolysis, averaging 3.3- and 2.0-fold higher in hemolyzed samples, respectively, indicating that (1) hemoglobin migrates into the alpha2-globulin region in this species and (2) notable intracellular potassium is released into plasma with hemolysis. Attempted conversion factors for compensation of hemolysis were considered inaccurate for 4 of 16 analytes due to non-significant regression lines. We also report that PCV provides an estimate of hemoglobin (g/L) using the formula: (2.59 × PCV) + 24.59. Given the spurious effects of hemolysis, the degree of this artifact should be reported with biochemistry data, and samples with moderate to severe hemolysis should be excluded from datasets when interpreting electrolyte, mineral, and protein results. This will ensure accurate data interpretation for individual turtles in rehabilitation or research settings and population-level data relevant to conservation-focused projects.

Characterizing stable isotope relationships between green turtle (Chelonia mydas) skin and unhatched eggs.

RATIONALE: Stable isotope analysis is used to understand the foraging habits and movements of a diverse set of organisms. Variability in stable isotope ratios among tissues derived from the same animal makes it difficult to compare data among study results in which different tissue types are evaluated. Isotopic relationships between two green turtle (Chelonia mydas) tissue types, skin and unhatched egg contents are unknown. Similarly, few data exist to evaluate the influence of time elapsed after oviposition (as a proxy for decomposition) on isotopic variability among unhatched eggs within the same nest. METHODS: Skin and unhatched egg contents were collected from 69 adult female green turtles and associated nests at the Archie Carr National Wildlife Refuge in Florida, USA. Values of δ13 C, δ15 N, and δ34 S were measured for both tissue types using a continuous flow isotope ratio mass spectrometer. Standardized major-axis (SMA) regression was used to generate conversion equations of carbon, nitrogen, and sulfur isotope ratios between the two tissue types. Model selection frameworks consisting of single-factor linear models were employed per isotope ratio to assess how egg time-in-nest affected intraclutch isotopic variability. RESULTS: Conversion equations for all three isotope ratios indicated significant relationships between skin and unhatched egg values, although model fits were lower than found in some studies examining similar patterns in other marine turtle species. The probability of increased intraclutch variability was significantly higher among eggs collected at longer intervals after deposition. CONCLUSIONS: This study reports the first-ever δ13 C and δ15 N conversion equations between skin and unhatched eggs for green turtles, and the first δ34 S conversion equation for any marine turtle species. SMA regression was used to directly convert tissue values bidirectionally, unlike equations generated using ordinary least-squares regression. Issues with increased intraclutch variability at later excavation dates highlight the importance of collecting unhatched eggs as soon as possible after hatchling emergence.