Piscichuvirus-Associated Severe Meningoencephalomyelitis in Aquatic Turtles, United States, 2009-2021.

Viruses from a new species of piscichuvirus were strongly associated with severe lymphocytic meningoencephalomyelitis in several free-ranging aquatic turtles from 3 coastal US states during 2009-2021. Sequencing identified 2 variants (freshwater turtle neural virus 1 [FTuNV1] and sea turtle neural virus 1 [STuNV1]) of the new piscichuvirus species in 3 turtles of 3 species. In situ hybridization localized viral mRNA to the inflamed region of the central nervous system in all 3 sequenced isolates and in 2 of 3 additional nonsequenced isolates. All 3 sequenced isolates phylogenetically clustered with other vertebrate chuvirids within the genus Piscichuvirus. FTuNV1 and STuNV1 shared ≈92% pairwise amino acid identity of the large protein, which narrowly places them within the same novel species. The in situ association of the piscichuviruses in 5 of 6 turtles (representing 3 genera) with lymphocytic meningoencephalomyelitis suggests that piscichuviruses are a likely cause of lymphocytic meningoencephalomyelitis in freshwater and marine turtles.

Identification of a novel Neorickettsia species in a Kemp's ridley sea turtle with granulomatous nephritis and development of a quantitative PCR assay.

An adult male Kemp's ridley turtle was found dead on the coast of Kenedy County, Texas, in August 2019 with bilateral severe, diffuse granulomatous nephritis. Pan-bacterial 16S rRNA gene polymerase chain reaction (PCR) and amplicon sequencing of affected tissue indicated the presence of a Neorickettsia. Neorickettsia is a genus of obligate intracellular Alphaproteobacteria that are transmitted by digenean trematodes. For further characterization, primers were designed to amplify and sequence the groEL gene. Phylogenetic analysis found that the organism was distinct from other known species to a degree consistent with a novel species. Immunohistochemistry using an antibody directed against a Neorickettsia surface protein showed bacterial clusters within the renal granulomas. A species-specific quantitative PCR was designed and detected the organism within the liver and colon of the index case. A quantitative PCR survey of grossly normal kidneys opportunistically collected from additional stranded sea turtle kidneys detected this organism in five of 15 Kemp's ridley turtles, two of nine green turtles, and neither of two loggerhead turtles. Recognition of this novel organism in an endangered species is concerning; additional work is underway to further characterize the potential of this organism as a pathogen of sea turtles.

Fibropapillomatosis and Chelonid Alphaherpesvirus 5 Infection in Kemp's Ridley Sea Turtles (Lepidochelys kempii).

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.

Plastic ingestion by green turtles (Chelonia mydas) over 33 years along the coast of Texas, USA.

Despite exponential growth of anthropogenic marine debris in recent decades, plastic ingestion by marine turtles in the Gulf of Mexico is not well understood. Gastrointestinal tracts were examined from 464 green turtles that stranded in Texas between 1987 and 2019, and 226 turtles ingested plastic (48.7%). This number doubled from 32.5% in 1987-1999 to 65.5% in 2019, but mass of ingested items was lowest in 2019. No turtles showed evidence of death directly related to plastic ingestion. Compared to other regions, plastic ingestion was low. Small turtles (<25 cm straight carapace length) ingested plastic more frequently and in greater amounts than larger turtles. Small turtles also ingested more hard plastic while larger turtles ingested more sheet-like and thread-like plastics, which may correspond to size-based habitat shifts. This is among the largest marine turtle ingestion studies to date and demonstrates an increasing prevalence of plastic ingestion.

Evolutionary Comparisons of Chelonid Alphaherpesvirus 5 (ChHV5) Genomes from Fibropapillomatosis-Afflicted Green (Chelonia mydas), Olive Ridley (Lepidochelys olivacea) and Kemp's Ridley (Lepidochelys kempii) Sea Turtles.

The spreading global sea turtle fibropapillomatosis (FP) epizootic is threatening some of Earth's ancient reptiles, adding to the plethora of threats faced by these keystone species. Understanding this neoplastic disease and its likely aetiological pathogen, chelonid alphaherpesvirus 5 (ChHV5), is crucial to understand how the disease impacts sea turtle populations and species and the future trajectory of disease incidence. We generated 20 ChHV5 genomes, from three sea turtle species, to better understand the viral variant diversity and gene evolution of this oncogenic virus. We revealed previously underappreciated genetic diversity within this virus (with an average of 2035 single nucleotide polymorphisms (SNPs), 1.54% of the ChHV5 genome) and identified genes under the strongest evolutionary pressure. Furthermore, we investigated the phylogeny of ChHV5 at both genome and gene level, confirming the propensity of the virus to be interspecific, with related variants able to infect multiple sea turtle species. Finally, we revealed unexpected intra-host diversity, with up to 0.15% of the viral genome varying between ChHV5 genomes isolated from different tumours concurrently arising within the same individual. These findings offer important insights into ChHV5 biology and provide genomic resources for this oncogenic virus.

Scavenging versus predation: shark-bite injuries in stranded sea turtles in the southeastern USA.

Injuries inflicted by sharks are a frequent observation in stranded sea turtles. Sharks prey on live turtles and scavenge carcasses, which can create uncertainty as to the cause of stranding when sea turtles are found dead with shark-bite wounds. Consequently, attributing the cause of stranding to a shark attack based purely on the presence of the characteristic wounds can overestimate predation by sharks as a cause of mortality. To better characterize the timing of shark-bite wounds relative to death of sea turtles in the southeastern USA, we performed necropsies on 70 stranded turtles that were found dead in which the predominant observation was bite wounds without any grossly evident vital responses (inflammation or healing). Postmortem examination included assessment for evidence of exsanguination and histopathological evaluation of skeletal muscle comprising wound margins. We characterized wounds as antemortem, perimortem, or postmortem based on specific criteria related to the presence or absence of supravital and intravital responses. Most (80%) shark-bite wounds were postmortem, 10% were antemortem, and 10% were perimortem. We found that antemortem and postmortem wounds were similar in extent and location except for wounds that primarily involved the shell, which were never found in cases of scavenging. For sea turtles found dead in the southeastern USA, our findings suggest that most shark-bite wounds without externally evident vital responses are due to scavenging. Additionally, this scavenging can significantly damage a carcass, potentially obscuring the detection of other causes of mortality. These findings should be considered when using data derived from stranded sea turtles to conduct mortality assessments.

Fibropapillomatosis prevalence and distribution in green turtles Chelonia mydas in Texas (USA).

Fibropapillomatosis (FP) is a neoplastic disease that can result in debilitating tumors in sea turtles. Initially identified in Florida, USA in 1937, it has since been detected in green turtle Chelonia mydas populations globally. FP was first identified and confirmed in Texas, USA in 2010. No FP tumors were documented in Texas prior to that year, though many green turtles were encountered and examined using standardized procedures since 1980. The present study was undertaken to identify temporal and spatial trends of FP prevalence in Texas since 2010. From 2010 through 2018, 1919 stranded or incidentally captured green turtles were documented with FP in Texas. FP prevalence was significantly correlated with year, hypothermic stunning, geographic region, and turtle size, as determined by logistic regression. FP was documented in <4.0% of the green turtles examined in Texas from 2010 to 2015, increasing to 21.6% in 2016, 27.3% in 2017, and 35.2% in 2018. More than twice as many hypothermic stunned green sea turtles had FP tumors as compared to those that were not hypothermic stunned. In Texas, FP was most prevalent in south Texas, particularly in the Laguna Madre, and associated channels. FP was more prevalent in turtles with straight carapace lengths 40.0-69.9 cm. The impact of this disease on green turtle population recovery in Texas is not yet apparent.

Brevetoxin exposure in sea turtles in south Texas (USA) during Karenia brevis red tide.

Five green (Chelonia mydas) and 11 Kemp's ridley (Lepidochelys kempii) sea turtles found dead, or that died soon after stranding, on the southern Texas (USA) coast during 2 Karenia brevis blooms (October 2015, September-October 2016) were tested for exposure to brevetoxins (PbTx). Tissues (liver, kidney) and digesta (stomach and intestinal contents) were analyzed by ELISA. Three green turtles found alive during the 2015 event and 2 Kemp's ridley turtles found alive during the 2016 event exhibited signs of PbTx exposure, including lethargy and/or convulsions of the head and neck. PbTx were detected in 1 or more tissues or digesta in all 16 stranded turtles. Detected PbTx concentrations ranged from 2 to >2000 ng g-1. Necropsy examination and results of PbTx analysis indicated that 10 of the Kemp's ridleys and 2 of the green turtles died from brevetoxicosis via ingestion. This is the first documentation of sea turtle mortality in Texas attributed to brevetoxicosis.

Sympatry or syntopy? Investigating drivers of distribution and co-occurrence for two imperiled sea turtle species in Gulf of Mexico neritic waters.

Animals co-occurring in a region (sympatry) may use the same habitat (syntopy) within that region. A central aim in ecology is determining what factors drive species distributions (i.e., abiotic conditions, dispersal limitations, and/or biotic interactions). Assessing the degree of biotic interactions can be difficult for species with wide ranges at sea. This study investigated the spatial ecology of two sea turtle species that forage on benthic invertebrates in neritic GoM waters: Kemp's ridleys (Lepidochelys kempii) and loggerheads (Caretta caretta). We used satellite tracking and modeled behavioral modes, then calculated individual home ranges, compared foraging areas, and determined extent of co-occurrence. Using six environmental variables and principal component analysis, we assessed similarity of chosen foraging sites. We predicted foraging location (eco-region) based on species, nesting site, and turtle size. For 127 turtles (64 Kemp's ridleys, 63 loggerheads) tracked from 1989 to 2013, foraging home ranges were nine to ten times larger for Kemp's ridleys than for loggerheads. Species intersected off all U.S. coasts and the Yucatán Peninsula, but co-occurrence areas were small compared to species' distributions. Kemp's ridley foraging home ranges were concentrated in the northern GoM, whereas those for loggerheads were concentrated in the eastern GoM. The two species were different in all habitat variables compared (latitude, longitude, distance to shore, net primary production, mean sea surface temperature, and bathymetry). Nesting site was the single dominant variable that dictated foraging ecoregion. Although Kemp's ridleys and loggerheads may compete for resources, the separation in foraging areas, significant differences in environmental conditions, and importance of nesting location on ecoregion selection (i.e., dispersal ability) indicate that adult females of these species do not interact greatly during foraging and that dispersal and environmental factors more strongly determine their distributions. These species show sympatry in this region but evidence for syntopy was rare.

Variability in age and size at maturation, reproductive longevity, and long-term growth dynamics for Kemp's ridley sea turtles in the Gulf of Mexico.

Effective management of protected sea turtle populations requires knowledge not only of mean values for demographic and life-history parameters, but also temporal and spatial trends, variability, and underlying causes. For endangered Kemp's ridley sea turtles (Lepidochelys kempii), the need for baseline information of this type has been emphasized during attempts to understand causes underlying the recent truncation in the recovery trajectory for nesting females. To provide insight into variability in age and size at sexual maturation (ASM and SSM) and long-term growth patterns likely to influence population trends, we conducted skeletochronological analysis of humerus bones from 333 Kemp's ridleys stranded throughout the Gulf of Mexico (GOM) from 1993 to 2010. Ranges of possible ASMs (6.8 to 21.8 yr) and SSMs (53.3 to 68.3 cm straightline carapace length (SCL)) estimated using the "rapprochement" skeletal growth mark associated with maturation were broad, supporting incorporation of a maturation schedule in Kemp's ridley population models. Mean ASMs estimated from rapprochement and by fitting logistic, generalized additive mixed, and von Bertalanffy growth models to age and growth data ranged from 11 to 13 yr; confidence intervals for the logistic model predicted maturation of 95% of the population between 11.9 and 14.8 yr. Early juvenile somatic growth rates in the GOM were greater than those previously reported for the Atlantic, indicating potential for differences in maturation trajectories between regions. Finally, long-term, significant decreases in somatic growth response were found for both juveniles and adults, which could influence recruitment to the reproductive population and observed nesting population trends.

Hypothermic stunning of green sea turtles in a western Gulf of Mexico foraging habitat.

Texas waters provide one of the most important developmental and foraging habitats for juvenile green turtles (Chelonia mydas) in the western Gulf of Mexico, but hypothermic stunning is a significant threat and was the largest cause of green turtle strandings in Texas from 1980 through 2015; of the 8,107 green turtles found stranded, 4,529 (55.9%) were victims of hypothermic stunning. Additionally, during this time, 203 hypothermic stunned green turtles were found incidentally captured due to power plant water intake entrapment. Overall, 63.9% of 4,529 hypothermic stunned turtles were found alive, and 92.0% of those survived rehabilitation and were released. Numbers of green turtles recorded as stranded and as affected by hypothermic stunning increased over time, and were most numerous from 2007 through 2015. Large hypothermic stunning events (with more than 450 turtles documented) occurred during the winters of 2009-2010, 2010-2011, 2013-2014, and 2014-2015. Hypothermic stunning was documented between November and March, but peaked at various times depending on passage of severe weather systems. Hypothermic stunning occurred state-wide, but was most prevalent in South Texas, particularly the Laguna Madre. In the Laguna Madre, hypothermic stunning was associated with an abrupt drop in water temperatures strong northerly winds, and a threshold mean water temperature of 8.0°C predicted large turtle hypothermic stunning events. Knowledge of environmental parameters contributing to hypothermic stunning and the temporal and spatial distribution of turtles affected in the past, can aid with formulation of proactive, targeted search and rescue efforts that can ultimately save the lives of many affected individuals, and aid with recovery efforts for this bi-national stock. Such rescue efforts are required under the U.S. Endangered Species Act and respond to humanitarian concerns of the public.

Inter-nesting movements and habitat-use of adult female Kemp's ridley turtles in the Gulf of Mexico.

Species vulnerability is increased when individuals congregate in restricted areas for breeding; yet, breeding habitats are not well defined for many marine species. Identification and quantification of these breeding habitats are essential to effective conservation. Satellite telemetry and switching state-space modeling (SSM) were used to define inter-nesting habitat of endangered Kemp's ridley turtles (Lepidochelys kempii) in the Gulf of Mexico. Turtles were outfitted with satellite transmitters after nesting at Padre Island National Seashore, Texas, USA, from 1998 through 2013 (n = 60); Rancho Nuevo, Tamaulipas, Mexico, during 2010 and 2011 (n = 11); and Tecolutla, Veracruz, Mexico, during 2012 and 2013 (n = 11). These sites span the range of nearly all nesting by this species. Inter-nesting habitat lies in a narrow band of nearshore western Gulf of Mexico waters in the USA and Mexico, with mean water depth of 14 to 19 m within a mean distance to shore of 6 to 11 km as estimated by 50% kernel density estimate, α-Hull, and minimum convex polygon methodologies. Turtles tracked during the inter-nesting period moved, on average, 17.5 km/day and a mean total distance of 398 km. Mean home ranges occupied were 725 to 2948 km2. Our results indicate that these nearshore western Gulf waters represent critical inter-nesting habitat for this species, where threats such as shrimp trawling and oil and gas platforms also occur. Up to half of all adult female Kemp's ridleys occupy this habitat for weeks to months during each nesting season. Because inter-nesting habitat for this species is concentrated in nearshore waters of the western Gulf of Mexico in both Mexico and the USA, international collaboration is needed to protect this essential habitat and the turtles occurring within it.

Foraging area fidelity for Kemp's ridleys in the Gulf of Mexico.

For many marine species, locations of key foraging areas are not well defined. We used satellite telemetry and switching state-space modeling (SSM) to identify distinct foraging areas used by Kemp's ridley turtles (Lepidochelys kempii) tagged after nesting during 1998-2011 at Padre Island National Seashore, Texas, USA (PAIS; N = 22), and Rancho Nuevo, Tamaulipas, Mexico (RN; N = 9). Overall, turtles traveled a mean distance of 793.1 km (±347.8 SD) to foraging sites, where 24 of 31 turtles showed foraging area fidelity (FAF) over time (N = 22 in USA, N = 2 in Mexico). Multiple turtles foraged along their migratory route, prior to arrival at their "final" foraging sites. We identified new foraging "hotspots" where adult female Kemp's ridley turtles spent 44% of their time during tracking (i.e., 2641/6009 tracking days in foraging mode). Nearshore Gulf of Mexico waters served as foraging habitat for all turtles tracked in this study; final foraging sites were located in water <68 m deep and a mean distance of 33.2 km (±25.3 SD) from the nearest mainland coast. Distance to release site, distance to mainland shore, annual mean sea surface temperature, bathymetry, and net primary production were significant predictors of sites where turtles spent large numbers of days in foraging mode. Spatial similarity of particular foraging sites selected by different turtles over the 13-year tracking period indicates that these areas represent critical foraging habitat, particularly in waters off Louisiana. Furthermore, the wide distribution of foraging sites indicates that a foraging corridor exists for Kemp's ridleys in the Gulf. Our results highlight the need for further study of environmental and bathymetric components of foraging sites and prey resources contained therein, as well as international cooperation to protect essential at-sea foraging habitats for this imperiled species.

Predicting the distribution of oceanic-stage Kemp's ridley sea turtles.

The inaccessibility of open ocean habitat and the cryptic nature of small animals are fundamental problems when assessing the distribution of oceanic-stage sea turtles and other marine animals sharing similar life-history traits. Most methods that estimate patterns of abundance cannot be applied in situations that are extremely data limited. Here, we use a movement ecology framework to generate the first predicted distributions for the oceanic stage of the Kemp's ridley sea turtle (Lepidochelys kempii). Our simulations of particle dispersal within ocean circulation models reveal substantial annual variation in distribution and survival among simulated cohorts. Such techniques can help prioritize areas for conservation, and supply inputs for more realistic demographic models attempting to characterize population trends.