Comparative plasma biochemistry analyte data in nesting leatherback (Dermochelys coriacea), foraging green (Chelonia mydas) and foraging and nesting hawksbill (Eretmochelys imbricata) sea turtles in Grenada, West Indies.

Blood biochemistry represents a minimally invasive tool for monitoring sea turtle health, assessing injured sea turtles and supporting conservation strategies. In Grenada, West Indies, plasma biochemical variables were examined in 33 nesting leatherback (Dermochelys coriacea), 49 foraging green (Chelonia mydas), 49 foraging hawksbill (Eretmochelys imbricata) and 12 nesting hawksbill sea turtles sampled between 2017 and 2022. Plasma biochemistry reference intervals are described herein except for nesting hawksbills, which are represented by descriptive statistics due to the low sample size. Select analyte concentrations were positively correlated with curved carapace length in leatherbacks (chloride), green turtles (total protein, albumin and globulin) and foraging hawksbills (total protein, albumin and phosphorus). Cholesterol (7.8 mmol/l ± 1.6 SD) and triglyceride (6.9 mmol/l ± 1.9 SD) concentrations were significantly higher in leatherbacks compared to foraging green turtles, foraging hawksbills and nesting hawksbills (P < 0.001 for all). Cholesterol was significantly higher in nesting hawksbills compared to foraging green turtles (P = 0.050) and foraging hawksbills (P = 0.050). Foraging hawksbills demonstrated significantly higher aspartate transaminase activities than leatherbacks (P = 0.002), green turtles (P = 0.009) and nesting hawksbills (P < 0.001). Biochemical results provide baseline population health data and support guidance for treatments during clinical sea turtle rehabilitation efforts. They also provide insight into species-specific physiologic differences and preludes further studies to better characterize the impacts of life-stage class on biochemistry reference intervals to better support wild sea turtle populations in Grenada.

Environmental and Nesting Variables Associated with Atlantic Leatherback Sea Turtle (Dermochelys coriacea) Embryonic and Hatching Success Rates in Grenada, West Indies.

Annual monitoring of leatherback sea turtle (Dermochelys coriacea) nesting grounds in Grenada, West Indies has identified relatively low hatch rates compared to worldwide trends. This study investigated the impact of selected environmental variables on leatherback sea turtle embryonic development and hatching success rates on Levera Beach in Grenada between 2015-2019. The mean number of nests per year and eggs per nest were 667.6 ± 361.6 and 80.7 ± 23.0 sd, respectively. Within excavated nests, 35.6% ± 22.0 sd of eggs successfully developed embryos and 30.6% ± 22.6 sd of eggs successfully hatched. The number of eggs per nest, along with embryo and hatching success rates, differed by nesting year. Embryo development success rate was associated with nest location, and both embryo development and hatching success rates were positively associated with nest depth and negatively associated with the percentage of eggs exhibiting microbial growth and with the presence of inspissated yolk. There was no embryo development or hatchling success association with month of the nesting season, distance from the high-water mark, distance from vegetation, nor maternal carapace length. The mean nest temperature was 31.7 °C ± 1.64 sd and mean temperatures during the middle third of egg incubation suggest clutches are highly skewed towards a preponderance of female hatchlings. Histopathologic findings in hatchling mortalities included severe, acute, multifocal, heterophilic bronchopneumonia with intralesional bacteria in 4/50 (8%) hatchlings. Data from this study guide conservation strategies by identifying risk factors and further avenues of research needed to support reproductive success of leatherback sea turtles in Grenada and the greater Caribbean region.

Kudoa hypoepicardialis and associated cardiac lesions in invasive red lionfish Pterois volitans in Grenada, West Indies.

Invasive red lionfish Pterois volitans (Linnaeus, 1758) represent an ongoing ecological threat within temperate and tropical waters. Relatively little is known regarding the overall health of P. volitans and their potential for spreading pathogens in non-native regions. Lionfish collected from inshore reefs of Grenada, West Indies, in 2019 and 2021 were identified as P. volitans based on cytochrome c oxidase subunit 1 barcoding. Gross and microscopic examination of tissues revealed myxozoan plasmodia in the hearts of 24/76 (31.6%) lionfish by histopathology or wet mount cytology. Further histopathologic examination revealed severe granulomatous inflammation and myofiber necrosis associated with developing plasmodia and presporogonic life stages. Fresh myxospores were morphologically and molecularly consistent with Kudoa hypoepicardialis, being quadrate in apical view with 4 valves and 4 equal polar capsules. The spore body was 5.1-7.9 (mean: 6.0) µm long, 8.1-9.8 (8.7) µm wide, and 6.9-8.5 (7.7) µm thick. Polar capsules were 2.3-2.7 (2.5) µm long and 0.9-1.6 (1.3) µm wide. 18S small subunit rDNA sequences were 99.81-99.87% similar to sequence data from the original description of the species. Novel 28S large subunit rDNA and elongation factor 2 data, which did not match any previously reported species, were provided. This is the first account of a myxozoan parasite of P. volitans, a new host record and locality for K. hypoepicardialis, and one of few reports describing pathogen-associated lesions in invasive lionfish.

Chelonid Alphaherpesvirus 5 Prevalence and First Confirmed Case of Sea Turtle Fibropapillomatosis in Grenada, West Indies.

Chelonid alphaherpesvirus 5 (ChHV5) is strongly associated with fibropapillomatosis, a neoplastic disease of sea turtles that can result in debilitation and mortality. The objectives of this study were to examine green (Chelonia mydas), hawksbill (Eretmochelys imbricata), and leatherback (Dermochelys coriacea) sea turtles in Grenada, West Indies, for fibropapillomatosis and to utilize ChHV5-specific PCR, degenerate herpesvirus PCR, and serology to non-invasively evaluate the prevalence of ChHV5 infection and exposure. One-hundred and sixty-seven turtles examined from 2017 to 2019 demonstrated no external fibropapilloma-like lesions and no amplification of ChHV5 DNA from whole blood or skin biopsies. An ELISA performed on serum detected ChHV5-specific IgY in 18/52 (34.6%) of green turtles tested. In 2020, an adult, female green turtle presented for necropsy from the inshore waters of Grenada with severe emaciation and cutaneous fibropapillomas. Multiple tumors tested positive for ChHV5 by qPCR, providing the first confirmed case of ChHV5-associated fibropapillomatosis in Grenada. These results indicate that active ChHV5 infection is rare, although viral exposure in green sea turtles is relatively high. The impact of fibropapillomatosis in Grenada is suggested to be low at the present time and further studies comparing host genetics and immunologic factors, as well as examination into extrinsic factors that may influence disease, are warranted.

Prevalence and phenotypic characterization of Salmonella enterica isolates from three species of wild marine turtles in Grenada, West Indies.

BACKGROUND AND AIM: Salmonella enterica causes enteric disease in mammals and may potentially be transmitted from marine turtles that shed the pathogen in the environment. Marine turtle-associated human salmonellosis is a potential public health concern in Grenada, as the island supports populations of leatherback turtles (Dermochelys coriacea), hawksbill turtles (Eretmochelys imbricata), and green turtles (Chelonia mydas) that interface with veterinarians and conservation workers, the local population, and the thousands of visitors that frequent the island yearly. To date, the prevalence of S. enterica has only been examined in a small subset of marine turtles in the Caribbean and no studies have been conducted in Grenada. The aim of this study was to quantify the prevalence of S. enterica in leatherback, hawksbill and green turtles in Grenada, characterize phenotypes and DNA profiles, and explore the potential risk to human health in the region. MATERIALS AND METHODS: A total of 102 cloacal swabs were obtained from nesting leatherback turtles and foraging hawksbill and green turtles. Samples were cultured on enrichment and selective media and isolates were phenotypically characterized using serotyping, pulsed-phase gel electrophoresis, and antibiotic susceptibility. Enrichment broths were additionally screened by polymerase chain reaction (PCR) using S. enterica-specific primers. RESULTS: S. enterica was cultured from 15/57 (26.3%) leatherback turtles, 0/28 hawksbill, and 0/17 green turtles. This included S. enterica serovars Montevideo, S. I:4,5,12:i:-, Salmonella Typhimurium, Salmonella Newport, S. I:6,7:-:-, and S. I:4,5,12:-:-. Five/15 leatherback turtles carried multiple serovars. Eight pulsotype groups were identified with multiple clustering; however, there was no clear association between pulsotype group and serotype profile. Five/71 isolates showed resistance to streptomycin or ampicillin. Twenty-one/57 leatherback turtles, 14/28 hawksbill turtles, and 8/17 green turtles tested positive for S. enterica by quantitative PCR. CONCLUSION: Nesting leatherback turtles actively shed S. enterica and poses a risk for zoonosis; however, the presence of viable pathogen in green and hawksbill species is unclear. These findings help elucidate the role of marine turtles as potential sources of zoonotic S. enterica and provide baseline data for one health research in Grenada and the wider Caribbean region.

Organic ultraviolet filters in nearshore waters and in the invasive lionfish (Pterois volitans) in Grenada, West Indies.

Sunscreens and other personal care products use organic ultraviolet (UV) filters such as oxybenzone, 4-methylbenzylidene camphor, Padimate-O, and octyl methoxycinnamate to prevent damage to human skin. While these compounds are effective at preventing sunburn, they have a demonstrated negative effect on cells and tissues across taxonomic levels. These compounds have a relatively short half-life in seawater but are continuously re-introduced via recreational activities and wastewater discharge, making them environmentally persistent. Because of this, testing seawater samples for the presence of these compounds may not be reflective of their abundance in the environment. Bioaccumulation of organic ultraviolet filters in a high-trophic level predator may provide greater insight to the presence and persistence of these compounds. To address this, the present study collected seawater samples as well as muscle and stomach content samples from the invasive Pacific lionfish (Pterois volitans) in the nearshore waters of Grenada, West Indies to examine the use of lionfish as potential bioindicator species. Seawater and lionfish samples were collected at four sites that are near point sources of wastewater discharge and that receive a high number of visitors each year. Samples were tested for the presence and concentrations of oxybenzone, 4-methylbenzylidene camphor (4-MBC), Padimate-O, and octyl methoxycinnamate (OMC) using liquid chromatography-mass spectrometry. Oxybenzone residues were detected in 60% of seawater samples and OMC residues were detected in 20% of seawater samples. Seawater samples collected in the surface waters near Grenada's main beach had oxybenzone concentrations more than ten times higher than seawater samples collected in less frequently visited areas and the highest prevalence of UV filters in lionfish. Residues of oxybenzone were detected in 35% of lionfish muscle and 4-MBC residues were detected in 12% of lionfish muscle. Padimate-O was not detected in either seawater or lionfish samples. No organic UV filters were detected in lionfish stomach contents. Histopathologic examination of lionfish demonstrated no significant findings attributed to UV filter toxicity. These findings report UV filter residue levels for the first time in inshore waters in Grenada. Results indicate that lionfish may be bioaccumulating residues and may be a useful sentinel model for monitoring organic ultraviolet filters in the Caribbean Sea.