Species and population specific gene expression in blood transcriptomes of marine turtles.

BACKGROUND: Transcriptomic data has demonstrated utility to advance the study of physiological diversity and organisms' responses to environmental stressors. However, a lack of genomic resources and challenges associated with collecting high-quality RNA can limit its application for many wild populations. Minimally invasive blood sampling combined with de novo transcriptomic approaches has great potential to alleviate these barriers. Here, we advance these goals for marine turtles by generating high quality de novo blood transcriptome assemblies to characterize functional diversity and compare global transcriptional profiles between tissues, species, and foraging aggregations. RESULTS: We generated high quality blood transcriptome assemblies for hawksbill (Eretmochelys imbricata), loggerhead (Caretta caretta), green (Chelonia mydas), and leatherback (Dermochelys coriacea) turtles. The functional diversity in assembled blood transcriptomes was comparable to those from more traditionally sampled tissues. A total of 31.3% of orthogroups identified were present in all four species, representing a core set of conserved genes expressed in blood and shared across marine turtle species. We observed strong species-specific expression of these genes, as well as distinct transcriptomic profiles between green turtle foraging aggregations that inhabit areas of greater or lesser anthropogenic disturbance. CONCLUSIONS: Obtaining global gene expression data through non-lethal, minimally invasive sampling can greatly expand the applications of RNA-sequencing in protected long-lived species such as marine turtles. The distinct differences in gene expression signatures between species and foraging aggregations provide insight into the functional genomics underlying the diversity in this ancient vertebrate lineage. The transcriptomic resources generated here can be used in further studies examining the evolutionary ecology and anthropogenic impacts on marine turtles.

Tropicalization of temperate ecosystems in North America: The northward range expansion of tropical organisms in response to warming winter temperatures.

Tropicalization is a term used to describe the transformation of temperate ecosystems by poleward-moving tropical organisms in response to warming temperatures. In North America, decreases in the frequency and intensity of extreme winter cold events are expected to allow the poleward range expansion of many cold-sensitive tropical organisms, sometimes at the expense of temperate organisms. Although ecologists have long noted the critical ecological role of winter cold temperature extremes in tropical-temperate transition zones, the ecological effects of extreme cold events have been understudied, and the influence of warming winter temperatures has too often been left out of climate change vulnerability assessments. Here, we examine the influence of extreme cold events on the northward range limits of a diverse group of tropical organisms, including terrestrial plants, coastal wetland plants, coastal fishes, sea turtles, terrestrial reptiles, amphibians, manatees, and insects. For these organisms, extreme cold events can lead to major physiological damage or landscape-scale mass mortality. Conversely, the absence of extreme cold events can foster population growth, range expansion, and ecological regime shifts. We discuss the effects of warming winters on species and ecosystems in tropical-temperate transition zones. In the 21st century, climate change-induced decreases in the frequency and intensity of extreme cold events are expected to facilitate the poleward range expansion of many tropical species. Our review highlights critical knowledge gaps for advancing understanding of the ecological implications of the tropicalization of temperate ecosystems in North America.

Hydrogen isotope assimilation and discrimination in green turtles.

Although hydrogen isotopes (δ2H) are commonly used as tracers of animal movement, minimal research has investigated the use of δ2H as a proxy to quantify resource and habitat use. While carbon and nitrogen are ultimately derived from a single source (food), the proportion of hydrogen in consumer tissues originates from two distinct sources: body water and food. Before hydrogen isotopes can be effectively used as a resource and habitat tracer, we need estimates of (net) discrimination factors (Δ2HNet) that account for the physiologically mediated differences in the δ2H values of animal tissues relative to that of the food and water sources they use to synthesize tissues. Here, we estimated Δ2HNet in captive green turtles (Chelonia mydas) by measuring the δ2H values of tissues (epidermis and blood components) and dietary macromolecules collected in two controlled feeding experiments. Tissue δ2H and Δ2HNet values varied systematically among tissues, with epidermis having higher δ2H and Δ2HNet values than blood components, which mirrors patterns between keratinaceous tissues (feathers, hair) and blood in birds and mammals. Serum/plasma of adult female green turtles had significantly lower δ2H values compared with juveniles, likely due to increased lipid mobilization associated with reproduction. This is the first study to quantify Δ2HNet values in a marine ectotherm, and we anticipate that our results will further refine the use of δ2H analysis to better understand animal resource and habitat use in marine ecosystems, especially coastal areas fueled by a combination of marine (e.g. micro/macroalgae and seagrass) and terrestrial (e.g. mangroves) primary production.

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.

Long-term trends in the foraging ecology and habitat use of an endangered species: an isotopic perspective.

Evaluating long-term drivers of foraging ecology and population productivity is crucial for providing ecological baselines and forecasting species responses to future environmental conditions. Here, we examine the trophic ecology and habitat use of North Atlantic leatherback turtles (St. Croix nesting population) and investigate the effects of large-scale oceanographic conditions on leatherback foraging dynamics. We used bulk and compound-specific nitrogen isotope analysis of amino acids (CSIA-AA) to estimate leatherback trophic position (TP) over an 18-year period, compare these estimates with TP estimates from a Pacific leatherback population, and elucidate the pre-nesting habitat use patterns of leatherbacks. Our secondary objective was to use oceanographic indices and nesting information from St. Croix leatherbacks to evaluate relationships between trophic ecology, nesting parameters, and regional environmental conditions measured by the North Atlantic Oscillation (NAO) and Atlantic Multidecadal Oscillation. We found no change in leatherback TP over time and no difference in TP between Atlantic and Pacific leatherbacks, indicating that differences in trophic ecology between populations are an unlikely driver of the population dichotomy between Pacific and Atlantic leatherbacks. Isotope data suggested that St. Croix leatherbacks inhabit multiple oceanic regions prior to nesting, although, like their conspecifics in the Pacific, individuals exhibit fidelity to specific foraging regions. Leatherback nesting parameters were weakly related to the NAO, which may suggest that positive NAO phases benefit St. Croix leatherbacks, potentially through increases in resource availability in their foraging areas. Our data contribute to the understanding of leatherback turtle ecology and potential mechanistic drivers of the dichotomy between populations of this protected species.

Intrapopulation variability in the timing of ontogenetic habitat shifts in sea turtles revealed using δ15 N values from bone growth rings.

Determining location and timing of ontogenetic shifts in the habitat use of highly migratory species, along with possible intrapopulation variation in these shifts, is essential for understanding mechanisms driving alternate life histories and assessing overall population trends. Measuring variations in multi-year habitat-use patterns is especially difficult for remote oceanic species. To investigate the potential for differential habitat use among migratory marine vertebrates, we measured the naturally occurring stable nitrogen isotope (δ15 N) patterns that differentiate distinct ocean regions to create a 'regional isotope characterization', analysed the δ15 N values from annual bone growth layer rings from dead-stranded animals, and then combined the bone and regional isotope data to track individual animal movement patterns over multiple years. We used humeri from juvenile North Pacific loggerhead turtles (Caretta caretta), animals that undergo long migrations across the North Pacific Ocean (NPO), using multiple discrete regions as they develop to adulthood. Typical of many migratory marine species, ontogenetic changes in habitat use throughout their decades-long juvenile stage is poorly understood, but each potential habitat has unique foraging opportunities and spatially explicit natural and anthropogenic threats that could affect key life-history parameters. We found a bimodal size/age distribution in the timing that juveniles underwent an ontogenetic habitat shift from the oceanic central North Pacific (CNP) to the neritic east Pacific region near the Baja California Peninsula (BCP) (42·7 ± 7·2 vs. 68·3 ± 3·4 cm carapace length, 7·5 ± 2·7 vs. 15·6 ± 1·7 years). Important to the survival of this population, these disparate habitats differ considerably in their food availability, energy requirements and threats, and these differences can influence life-history parameters such as growth, survival and future fecundity. This is the first evidence of alternative ontogenetic shifts and habitat-use patterns for juveniles foraging in the eastern NPO. We combine two techniques, skeletochronology and stable isotope analysis, to reconstruct multi-year habitat-use patterns of a remote migratory species, linked to estimated ages and body sizes of individuals, to reveal variable ontogeny during the juvenile life stage that could drive alternate life histories and that has the potential to illuminate the migration patterns for other species with accretionary tissues.

Stable isotope discrimination factors and between-tissue isotope comparisons for bone and skin from captive and wild green sea turtles (Chelonia mydas).

RATIONALE: The ecological application of stable isotope analysis (SIA) relies on taxa- and tissue-specific stable carbon (Δ13 C) and nitrogen (Δ15 N) isotope discrimination factors, determined with captive animals reared on known diets for sufficient time to reflect dietary isotope ratios. However, captive studies often prohibit lethal sampling, are difficult with endangered species, and reflect conditions not experienced in the wild. METHODS: We overcame these constraints and determined the Δ13 C and Δ15 N values for skin and cortical bone from green sea turtles (Chelonia mydas) that died in captivity and evaluated the utility of a mathematical approach to predict discrimination factors. Using stable carbon (δ13 C values) and nitrogen (δ15 N values) isotope ratios from captive and wild turtles, we established relationships between bone stable isotope (SI) ratios and those from skin, a non-lethally sampled tissue, to facilitate comparisons of SI ratios among studies using multiple tissues. RESULTS: The mean (±SD) Δ13 C and Δ15 N values (‰) between skin and bone from captive turtles and their diet (non-lipid-extracted) were 2.3 ± 0.3 and 4.1 ± 0.4 and 2.1 ± 0.6 and 5.1 ± 1.1, respectively. The mathematically predicted Δ13 C and Δ15 N values were similar (to within 1‰) to the experimentally derived values. The mean δ15 N values from bone were higher than those from skin for captive (+1.0 ± 0.9‰) and wild (+0.8 ± 1.0‰) turtles; the mean δ13 C values from bone were lower than those from skin for wild turtles (-0.6 ± 0.9‰), but the same as for captive turtles. We used linear regression equations to describe bone vs skin relationships and create bone-to-skin isotope conversion equations. CONCLUSIONS: For sea turtles, we provide the first (a) bone-diet SI discrimination factors, (b) comparison of SI ratios from individual-specific bone and skin, and (c) evaluation of the application of a mathematical approach to predict stable isotope discrimination factors. Our approach opens the door for future studies comparing different tissues, and relating SI ratios of captive to wild animals.

Growth dynamics of juvenile loggerhead sea turtles undergoing an ontogenetic habitat shift.

Ontogenetic niche theory predicts that individuals may undergo one or more changes in habitat or diet throughout their lifetime to maintain optimal growth rates, or to optimize trade-offs between mortality risk and growth. We combine skeletochronological and stable nitrogen isotope (δ15N) analyses of sea turtle humeri (n = 61) to characterize the growth dynamics of juvenile loggerhead sea turtles (Caretta caretta) during an oceanic-to-neritic ontogenetic shift. The primary objective of this study was to determine how ontogenetic niche theory extends to sea turtles, and to individuals with different patterns of resource use (discrete shifters, n = 23; facultative shifters n = 14; non-shifters, n = 24). Mean growth rates peaked at the start of the ontogenetic shift (based on change in δ15N values), but returned to pre-shift levels within 2 years. Turtles generally only experienced 1 year of relatively high growth, but the timing of peak growth relative to the start of an ontogenetic shift varied among individuals (before, n = 14; during, n = 12; after, n = 8). Furthermore, no reduction in growth preceded the transition, as is predicted by ontogenetic niche theory. Annual growth rates were similar between non-transitioning turtles resident in oceanic and neritic habitats and turtles displaying alternative patterns of resource use. These results suggest that factors other than maximization of size-specific growth may more strongly influence the timing of ontogenetic shifts in loggerhead sea turtles, and that alternative patterns of resource use may have limited influence on somatic growth and age at maturation in this species.

VALIDATION OF ULTRASOUND AS A NONINVASIVE TOOL TO MEASURE SUBCUTANEOUS FAT DEPTH IN LEATHERBACK SEA TURTLES (DERMOCHELYS CORIACEA).

Leatherback turtles (Dermochelys coriacea) undergo substantial cyclical changes in body condition between foraging and nesting. Ultrasonography has been used to measure subcutaneous fat as an indicator of body condition in many species but has not been applied in sea turtles. To validate this technique in leatherback turtles, ultrasound images were obtained from 36 live-captured and dead-stranded immature and adult turtles from foraging and nesting areas in the Pacific and Atlantic oceans. Ultrasound measurements were compared with direct measurements from surgical biopsy or necropsy. Tissue architecture was confirmed histologically in a subset of turtles. The dorsal shoulder region provided the best site for differentiation of tissues. Maximum fat depth values with the front flipper in a neutral (45-90°) position demonstrated good correlation with direct measurements. Ultrasound-derived fat measurements may be used in the future for quantitative assessment of body condition as an index of health in this critically endangered species.

Effects of demineralization on the stable isotope analysis of bone samples.

RATIONALE: The sampling of sequential, annually formed bone growth layers for stable carbon (δ(13)C values) and nitrogen (δ(15)N values) isotope analysis (SIA) can provide a time series of foraging ecology data. To date, no standard protocol exists for the pre-SIA treatment of cortical samples taken from fresh, modern, bones. METHODS: Based on the SIA of historical bone, it is assumed that fresh bone samples must be pre-treated with acid prior to SIA. Using an elemental analyzer coupled to an isotope ratio mass spectrometer to measure stable carbon and nitrogen ratios, we tested the need to acidify cortical bone powder with 0.25 M HCl prior to SIA to isolate bone collagen for the determination of δ(13)C and δ(15)N values. We also examined the need for lipid extraction to remove potential biases related to δ(13)C analysis, based on a C:N ratio threshold of 3.5. RESULTS: It was found that acidification of micromilled cortical bone samples from marine turtles does not affect their δ(15)N values, and the small effect acidification has on δ(13)C values can be mathematically corrected for, thus eliminating the need for pre-SIA acidification of cortical bone. The lipid content of the cortical bone samples was low, as measured by their C:N ratios, indicating that lipid extracting cortical bone samples from modern marine turtles is unnecessary. CONCLUSIONS: We present a standard protocol for testing fresh, modern cortical bone samples prior to SIA, facilitating direct comparison of future studies. Based on the results obtained from marine turtle bones, pre-acidification and lipid removal of cortical bone are not recommended. This is especially useful as there is frequently not enough bone material removed via micromilling of sequential growth layers to accommodate both acid treatment and SIA.

Age and residency duration of loggerhead turtles at a North Pacific bycatch hotspot using skeletochronology.

For migratory marine animals, like sea turtles, effective conservation can be challenging because key demographic information such as duration of life stages and exposure to spatially explicit threats in different habitats are often unknown. In the eastern Pacific near the Baja California Peninsula (BCP), Mexico, tens of thousands of endangered North Pacific loggerhead sea turtles (Caretta caretta) concentrate at a foraging area known to have high rates of fishery bycatch. Because stage survivorship of loggerheads in the BCP will vary significantly depending on the number of years spent in this region, we applied skeletochronology to empirically estimate residency duration in this loggerhead hotspot. The observed age distribution obtained from skeletochronology analysis of 146 dead-stranded loggerheads ranged from three to 24 years old, suggesting a BCP residency of >20 years. Given the maximum estimated age and a one-year migration to western Pacific nesting beaches, we infer age-at-maturation for BCP loggerheads at ~25 years old. We also examine survivorship at varying BCP residency durations by applying our findings to current annual mortality estimates. Predicted survivorship of loggerheads spending over 20 years in this BCP foraging habitat is less than 10%, and given that ~43,000 loggerhead turtles forage here, a significant number of turtles are at extreme risk in this region. This is the first empirical evidence supporting estimated age-at-maturation for BCP North Pacific loggerheads, and the first estimates of BCP stage survivorship. Our findings emphasize the urgent need for continued and effective international conservation efforts to minimize bycatch of this endangered species.

Predicting bycatch hotspots for endangered leatherback turtles on longlines in the Pacific Ocean.

Fisheries bycatch is a critical source of mortality for rapidly declining populations of leatherback turtles, Dermochelys coriacea. We integrated use-intensity distributions for 135 satellite-tracked adult turtles with longline fishing effort to estimate predicted bycatch risk over space and time in the Pacific Ocean. Areas of predicted bycatch risk did not overlap for eastern and western Pacific nesting populations, warranting their consideration as distinct management units with respect to fisheries bycatch. For western Pacific nesting populations, we identified several areas of high risk in the north and central Pacific, but greatest risk was adjacent to primary nesting beaches in tropical seas of Indo-Pacific islands, largely confined to several exclusive economic zones under the jurisdiction of national authorities. For eastern Pacific nesting populations, we identified moderate risk associated with migrations to nesting beaches, but the greatest risk was in the South Pacific Gyre, a broad pelagic zone outside national waters where management is currently lacking and may prove difficult to implement. Efforts should focus on these predicted hotspots to develop more targeted management approaches to alleviate leatherback bycatch.

The urban lives of green sea turtles: Insights into behavior in an industrialized habitat using an animal-borne camera.

The cryptic and aquatic life histories of sea turtles have made them a challenging group to directly observe, leaving significant knowledge gaps regarding social behavior and fine-scale elements of habitat use. Using a custom-designed animal-borne camera, we observed previously undocumented behaviors by green turtles (Chelonia mydas) at a foraging area in San Diego Bay, a highly urbanized ecosystem in California, USA. We deployed a suction-cup-attached pop-off camera (manufactured by Customized Animal Tracking Solutions) on 11 turtles (mean straight carapace length = 84.0 ± 11.2 cm) for between 1 and 30.8 h. Video recordings, limited to sunlit hours, provided 73 h of total observation time between May 2022 and June 2023. We observed 32 conspecific interactions; we classified 18 as active, entailing clear social behaviors, as compared with 14 passive interactions representing brief, chance encounters. There was no evidence for agonistic interactions. The camera additionally revealed that green turtles consistently use metal structures within urban San Diego Bay. In seven instances, turtles exhibited rubbing behavior against metal structures, and we observed two examples of turtles congregating at these structures. High rates of intraspecific interaction exhibited relatively consistently among individuals provide a compelling case for sociality for green turtles in San Diego Bay, adding to a growing research base updating their historical label of "non-social." The frequent use of metal structures by the population, in particular the rubbing of exposed skin, has implications for behavioral adaptations to urban environments. Our study exemplifies the promise of technological advances (e.g., underwater and animal-borne cameras) for updating natural history paradigms, even for well-studied populations.

Long-term changes in body size of green turtles nesting on Trindade Island, Brazil: Signs of recovery?

Trindade Island is an important wildlife refuge in the South Atlantic Ocean and hosts the largest nesting population of green turtles (Chelonia mydas) in Brazil, about which temporal ecological dynamics are still not well understood. The present study examines 23 years of nesting for green turtles at this remote island to evaluate annual mean nesting size (MNS) changes and post-maturity somatic growth rates. Our results show a significant decrease in annual MNS over the study; Whereas MNS during the first three consecutively monitored years (1993-1995) was 115.1 ± 5.4 cm, during the last three years (2014-2016) it was 111.2 ± 6.3 cm. There was no significant change in post-maturity somatic growth rate over the course of the study; the mean annual growth rate was 0.25 ± 0.62 cm/year. These findings suggest an increase in the relative proportion of smaller, presumptive neophyte nesters appearing in Trindade during the study period.

Anthropogenic uranium signatures in turtles, tortoises, and sea turtles from nuclear sites.

Chelonians (turtles, tortoises, and sea turtles) grow scute keratin in sequential layers over time. Once formed, scute keratin acts as an inert reservoir of environmental information. For chelonians inhabiting areas with legacy or modern nuclear activities, their scute has the potential to act as a time-stamped record of radionuclide contamination in the environment. Here, we measure bulk (i.e. homogenized scute) and sequential samples of chelonian scute from the Republic of the Marshall Islands and throughout the United States of America, including at the Barry M. Goldwater Air Force Range, southwestern Utah, the Savannah River Site, and the Oak Ridge Reservation. We identify legacy uranium (235U and 236U) contamination in bulk and sequential chelonian scute that matches known nuclear histories at these locations during the 20th century. Our results confirm that chelonians bioaccumulate uranium radionuclides and do so sequentially over time. This technique provides both a time series approach for reconstructing nuclear histories from significant past and present contexts throughout the world and the ability to use chelonians for long-term environmental monitoring programs (e.g. sea turtles at Enewetok and Bikini Atolls in the Republic of the Marshall Islands and in Japan near the Fukushima Daiichi reactors).

Shifting the life-history paradigm: discovery of novel habitat use by hawksbill turtles.

Adult hawksbill turtles (Eretmochelys imbricata) are typically described as open-coast, coral reef and hard substrate dwellers. Here, we report new satellite tracking data on female hawksbills from several countries in the eastern Pacific that revealed previously undocumented behaviour for adults of the species. In contrast to patterns of habitat use exhibited by their Caribbean and Indo-Pacific counterparts, eastern Pacific hawksbills generally occupied inshore estuaries, wherein they had strong associations with mangrove saltwater forests. The use of inshore habitats and affinities with mangrove saltwater forests presents a previously unknown life-history paradigm for adult hawksbill turtles and suggests a potentially unique evolutionary trajectory for the species. Our findings highlight the variability in life-history strategies that marine turtles and other wide-ranging marine wildlife may exhibit among ocean regions, and the importance of understanding such disparities from an ecological and management perspective.

Age-specific growth and maturity estimates for the flatback sea turtle (Natator depressus) by skeletochronology.

To address a major knowledge gap for flatback sea turtles (Natator depressus), a species endemic to Australia and considered 'Data Deficient' for IUCN Red List assessment, we present the first-ever skeletochronology-derived age and growth rate estimates for this species. Using a rare collection of bone samples gathered from across northern Australia, we applied skeletochronology and characterized the length-at-age relationship, established baseline growth rates from the hatchling to adult life stages, and produced empirical estimates of age-at- and size-at-sexual-maturation (ASM, SSM). We analyzed humeri from 74 flatback sea turtles ranging in body size from 6.0-96.0 cm curved carapace length (CCL), and recovered from Western Australia (n = 48), Eastern Australia (n = 13), central Australia (n = 8; Northern Territory n = 3, the Gulf of Carpentaria n = 5), and unknown locations (n = 5). We identified the onset of sexual maturity for 29 turtles, based on rapprochement growth patterns in the bones. Estimates for ASM ranged from 12.0 to 23.0 years (mean: 16.3 ± 0.53 SE), SSM ranged from 76.1 to 94.0 cm CCL (mean: 84.9 ± 0.90 SE), and maximum observed reproductive longevity was 31 years for a 45-year old male flatback. Growth was modeled as a smoothing spline fit to the size-at-age relationship and at the mean SSM (84.9 cm CCL) corresponded with a spline-predicted maturity age of 18 years (95% CI: 16 to 24), while mean nesting sizes reported in the literature (86.4 to 94 cm CCL) corresponded to estimated ages of 24+ years. A bootstrapped von Bertalanffy growth model was also applied and showed consistencies with the spline curve, yielding an estimated upper size limit, Linf, at 89.2 ± 0.04 cm (95% CI: 85.5 to 95.9 cm) with the intrinsic growth rate parameter, k, at 0.185 ± 0.0004 (0.16 to 0.22); at the same mean SSM (84.9 cm CCL) the estimated ASM was 16.3 ± 0.05 years (95% CI: 12.8 to 27.7 years). Lastly, four of the samples analyzed were collected from deceased adult females that had previous sizes known from on-going mark/recapture studies at nesting sites in Western Australia. The paired CCL data (measured at nesting and back-calculated) did not significantly differ (p = 0.875). This first skeletochronology study for flatback sea turtles generates valuable empirical estimates for ongoing conservation and management efforts.

PLASMA BIOCHEMISTRY PROFILES OF JUVENILE GREEN TURTLES (CHELONIA MYDAS) FROM THE BAHAMAS WITH A POTENTIAL INFLUENCE OF DIET.

Plasma biochemistry profiles aid health assessment of marine turtles, but knowledge of the influence of regional biological factors (e.g., habitat, diet) on marine turtle blood plasma values is limited. To investigate the influence of diet on plasma biochemistry values in juvenile green turtles (Chelonia mydas), we used carbon and nitrogen stable isotopes to provide a quantitative estimate of forage items in green turtles feeding at two distinct areas (Bonefish Hole and South Flats) in Bimini, Bahamas. Plasma samples were obtained from 13 turtles in Bonefish Hole (a mangrove tidal estuary) and 15 turtles in South Flats (an open water seagrass bed) in 2018. All turtles appeared outwardly healthy. Sessile filter feeders contributed the largest proportion of diet in Bonefish Hole, and seagrass contributed the highest proportion of diet in South Flats. Turtles at Bonefish Hole presented significantly lower cholesterol, total protein, phosphorus, triglycerides, and aspartate transaminase compared to South Flats. Across all turtles, those feeding primarily on red algae presented the highest uric acid and alkaline phosphatase, and turtles with a seagrass-dominated diet had the highest cholesterol. Understanding dietary influence on plasma biochemistry may help explain variances seen in local health and nutritional evaluations, and the trends reported can aid the interpretation of plasma analyte values in marine turtles.

Quantifying local ecological knowledge to model historical abundance of long-lived, heavily-exploited fauna.

Deriving robust historical population trends for long-lived species subject to human exploitation is challenging in scenarios where long-term scientific data are scarce or unavailable, as often occurs for species affected by small-scale fisheries and subsistence hunting. The importance of Local Ecological Knowledge (LEK) in data-poor scenarios is increasingly recognized in conservation, both in terms of uncovering historical trends and for engaging community stewardship of historic information. Building on previous work in marine historical ecology and local ecological knowledge, we propose a mixed socio-ecological framework to reliably document and quantify LEK to reconstruct historical population trends. Our method can be adapted by interdisciplinary teams to study various long-lived taxa with a history of human use. We demonstrate the validity of our approach by reconstructing long-term abundance data for the heavily-exploited East Pacific green turtle (Chelonia mydas) in Baja California, Mexico, which was driven to near extinction by a largely unregulated fishery from the early 1950s to the 1980s. No scientific baseline abundance data were available for this time-frame because recent biological surveys started in 1995 after all green turtle fisheries in the area were closed. To fill this data gap, we documented LEK among local fishers using ethnographic methods and obtained verified, qualitative data to understand the socio-environmental complexity of the green turtle fishery. We then established an iterative framework to synthesize and quantify LEK using generalized linear models (GLMs) and nonlinear regression (NLR) to generate a standardized, LEK-derived catch-per-unit-effort (CPUE) time-series. CPUE is an index of abundance that is compatible with contemporary scientific survey data. We confirmed the accuracy of LEK-derived CPUE estimates via comparisons with fisheries statistics available for 1962-1982. We then modeled LEK-derived abundance trends prior to 1995 using NLR. Our model established baseline abundance and described historical declines, revealing that the most critical (exponential) decline occurred between 1960 and 1980. This robust integration of LEK data with ecological science is of critical value for conservation and management, as it contributes to a holistic view of a species' historic and contemporary conservation status.

Persistent organic pollutants in green sea turtles (Chelonia mydas) inhabiting two urbanized Southern California habitats.

Within Southern California, east Pacific green sea turtles (Chelonia mydas) forage year-round, taking advantage of diverse food resources, including seagrass, marine algae, and invertebrates. Assessing persistent organic pollutants (POP) in green turtle aggregations in the Seal Beach National Wildlife Refuge (SBNWR, n = 17) and San Diego Bay (SDB, n = 25) can help quantify contamination risks for these populations. Blood plasma was analyzed for polychlorinated biphenyls (PCBs), organochlorinated pesticides (OCPs), and polybrominated diphenyl ethers (PBDEs). PCBs and body size explained much of the separation of turtles by foraging aggregation in a principal component analysis. Turtles from SDB had significantly (p < 0.001) higher total PCBs than SBNWR turtles. Most PCBs detected in turtles were non-dioxin-like PCB congeners (153, 138, 99) that are associated with neurotoxicity. Recaptured turtles' POP levels changed significantly over time indicating significant variation in POP levels through time and space, even among adjacent foraging locations.