Frequency-dependent temporary threshold shifts in the Eastern painted turtle (Chrysemys picta picta).

Testudines are a highly threatened group facing an array of stressors, including alteration of their sensory environment. Underwater noise pollution has the potential to induce hearing loss and disrupt detection of biologically important acoustic cues and signals. To examine the conditions that induce temporary threshold shifts (TTS) in hearing in the freshwater Eastern painted turtle (Chrysemys picta picta), three individuals were exposed to band limited continuous white noise (50-1000 Hz) of varying durations and amplitudes (sound exposure levels ranged from 151 to 171 dB re 1 µPa2 s). Control and post-exposure auditory thresholds were measured and compared at 400 and 600 Hz using auditory evoked potential methods. TTS occurred in all individuals at both test frequencies, with shifts of 6.1-41.4 dB. While the numbers of TTS occurrences were equal between frequencies, greater shifts were observed at 600 Hz, a frequency of higher auditory sensitivity, compared to 400 Hz. The onset of TTS occurred at 154 dB re 1 µPa2 s for 600 Hz, compared to 158 dB re 1 µPa2 s at 400 Hz. The 400-Hz onset and patterns of TTS growth and recovery were similar to those observed in previously studied Trachemys scripta elegans, suggesting TTS may be comparable across Emydidae species.

Home range of three turtle species in Central Yucatan. A comparative study.

Home range is a fundamental characteristic of an animal natural history. The study of home range provides information on the sites where organisms forage for food, find shelter, or locate mates. Home range size and shape can change throughout the lifespan of an organism, during the year, or across seasons, driven by resource availability and the basic needs for each organism. For freshwater and semi-aquatic turtles, home range is greatly affected by water availability, humidity, and temperature throughout the year, nevertheless demographic factors such age and sex are also important determinants of home range size. In this study we estimated home range and dispersal movements for Kinosternon creaseri, Terrapene yucatana, and Rhinoclemmys areolata in a semi-tropical dry forest in central Yucatán. For a two-year period, turtles were surveyed using hoop traps and visual encounters. Twenty-one individuals (5-8 per species) were equipped with radio transmitters to track them across the landscape. Distances between relocations and home range were compared across species seasons, sex, and interactions of these variables. Monthly average movements were positively correlated with rain in the three species studied. Home range of R. areolata was larger than those of K. creaseri and T. yucatana. Home range of the three studied species were larger during the wet season. Home range overlap index within same species individuals was higher during the rainy than dry season, but overall overlap is low between and within species.

Nanoplastic pollution changes the intestinal microbiome but not the morphology or behavior of a freshwater turtle.

Humans produce 350 million metric tons of plastic waste per year, leading to microplastic pollution and widespread environmental contamination, particularly in aquatic environments. This subsequently impacts aquatic organisms in myriad ways, yet the vast majority of research is conducted in marine, rather than freshwater systems. In this study, we exposed eggs and hatchlings of the Chinese soft-shelled turtle (Pelodiscus sinensis) to 80-nm polystyrene nanoplastics (PS-NPs) and monitored the impacts on development, behavior and the gut microbiome. We demonstrate that 80-nm PS-NPs can penetrate the eggshell and move into developing embryos. This led to metabolic impairments, as evidenced by bradycardia (a decreased heart rate), which persisted until hatching. We found no evidence that nanoplastic exposure affected hatchling morphology, growth rates, or levels of boldness and exploration, yet we discuss some potential caveats here. Exposure to nanoplastics reduced the diversity and homogeneity of gut microbiota in P. sinensis, with the level of disruption correlating to the length of environmental exposure (during incubation only or post-hatching also). Thirteen core genera (with an initial abundance >1 %) shifted after nanoplastic treatment: pathogenic bacteria increased, beneficial probiotic bacteria decreased, and there was an increase in the proportion of negative correlations between bacterial genera. These changes could have profound impacts on the viability of turtles throughout their lives. Our study highlights the toxicity of environmental NPs to the embryonic development and survival of freshwater turtles. We provide insights about population trends of P. sinensis in the wild, and future directions for research.

Alfaxalone is an effective anesthetic for the electrophysiological study of anoxia-tolerance mechanisms in western painted turtle pyramidal neurons.

Anoxia in the mammalian brain leads to hyper-excitability and cell death; however, this cascade of events does not occur in the anoxia-tolerant brain of the western painted turtle, Chrysemys picta belli. The painted turtle has become an important anoxia-tolerant model to study brain, heart, and liver function in the absence of oxygen, but being anoxia-tolerant likely means that decapitation alone is not a suitable method of euthanasia. Many anesthetics have long-term effects on ion channels and are not appropriate for same day experimentation. Using whole-cell electrophysiological techniques, we examine the effects of the anesthetic, Alfaxalone, on pyramidal cell action potential amplitude, threshold, rise and decay time, width, frequency, whole cell conductance, and evoked GABAA receptors currents to determine if any of these characteristics are altered with the use of Alfaxalone for animal sedation. We find that Alfaxalone has no long-term impact on action potential parameters or whole-cell conductance. When acutely applied to naïve tissue, Alfaxalone did lengthen GABAA receptor current decay rates by 1.5-fold. Following whole-animal sedation with Alfaxalone, evoked whole cell GABAA receptor current decay rates displayed an increasing trend with 1 and 2 hours after brain sheet preparation, but showed no significant change after a 3-hour washout period. Therefore, we conclude that Alfaxalone is a suitable anesthetic for same day use in electrophysiological studies in western painted turtle brain tissue.

Morphological variations and demographic responses of the Mediterranean pond turtle Mauremys leprosa to heterogeneous aquatic habitats.

Human activities affect terrestrial and aquatic habitats leading to changes at both individual and population levels in wild animal species. In this study, we investigated the phenotype and demographics of the Mediterranean pond turtle Mauremys leprosa (Schweigger, 1812) in contrasted environments of Southern France: two peri-urban rivers receiving effluents from wastewater treatment plants (WWTP), and another one without sewage treatment plant. Our findings revealed the presence of pesticides and pharmaceuticals in the three rivers of investigation, the highest diversities and concentrations of pollutants being found in the river subsections impacted by WWTP effluents. Principal component analysis and hierarchical clustering identified three levels of habitat quality, with different pollutant concentrations, thermal conditions, nutrient, and organic matter levels. The highest turtle densities, growth rates, and body sizes were estimated in the most disturbed habitats, suggesting potential adult benefits derived from harsh environmental conditions induced by pollution and eutrophication. Conversely, juveniles were the most abundant in the least polluted habitats, suggesting adverse effects of pollution on juvenile survival or adult reproduction. This study suggests that turtles living in polluted habitats may benefit from enhanced growth and body size, at the expense of reproductive success.

Dynamic shoreline alterations and their impacts on Olive Ridley Turtle (Lepidochelys olivacea) nesting sites in Gahirmatha Marine Wildlife Sanctuary, Odisha (India).

Currently, sea turtle habitats are being altered by climate change and human activities, with habitat loss posing an urgent threat to Indian sea turtles. Thus, the objective of this study is to analyze the dynamic shoreline alterations and their impacts on Olive Ridley Sea Turtle (ORT) nesting sites in Gahirmatha Marine Wildlife Sanctuary from 1990 to 2022. Landsat satellite images served as input datasets to assess dynamic shoreline changes. This study assessed shoreline alterations and their rates across 929 transects divided into four zones using the Digital Shoreline Analysis System (DSAS) software. The results revealed a significant 14-km northward shift in the nesting site due to substantial coastal erosion, threatening the turtles' Arribada. This study underscores the need for conservation efforts to preserve nesting environments amidst changing coastal landscapes, offering novel insights into the interaction between coastal processes and marine turtle nesting behaviors.

How turtles keep their cool: Seasonal and diel basking patterns in a tropical turtle.

Behavioural thermoregulation by ectotherms is an important mechanism for maintaining body temperatures to optimise physiological performance. Experimental studies suggest that nocturnal basking by Krefft's river turtles (Emydura macquarii krefftii) in the tropics may allow them to avoid high water temperatures, however, this hypothesis has yet to be tested in the field. In this study, we examined the influence of environmental temperature on seasonal and diel patterns of basking in E. m. krefftii in tropical north Queensland, Australia. Wildlife cameras were used to document turtle basking events for seven consecutive days and nights for each month over a year (April 2020-March 2021). Air and water temperatures were recorded simultaneously using temperature loggers. We used a negative binomial mixed effects model to compare mean basking durations (min) occurring among four environmental temperature categories based on population thermal preference (26 °C): 1) air temperature above and water temperature below preferred temperature; 2) air temperature below and water temperature above preferred temperature; 3) air and water temperatures both above preferred temperature; and 4) air and water temperatures both below preferred temperature. Basking behaviour was influenced significantly by the relationship between air and water temperature. During the day, turtles spent significantly less time basking when both air and water temperatures were above their preferred temperatures. Conversely, at night, turtles spent significantly more time basking when water temperatures were warm and air temperatures were cool relative to their preferred temperature. This study adds to the growing body of work indicating pronounced heat avoidance as a thermoregulatory strategy among tropical reptile populations.

Trained quantity discrimination in invasive red-eared slider and a comparison with the native stripe-necked turtle.

Little is known about the behavioral and cognitive traits that best predict invasion success. Evidence is mounting that cognitive performance correlates with survival and fecundity, two pivotal factors for the successful establishment of invasive populations. We assessed the quantity discrimination ability of the globally invasive red-eared slider (Trachemys scripta elegans). We further compared it to that of the native stripe-necked turtle (Mauremys sinensis), which has been previously evaluated for its superior quantity discrimination ability. Specifically, our experimental designs aimed to quantify the learning ability as numerosity pairs increased in difficulty (termed fixed numerosity tests), and the immediate response when turtles were presented with varied challenges concurrently in the same tests (termed mixed numerosity tests). Our findings reaffirm the remarkable ability of freshwater turtles to discern numerical differences as close as 9 vs 10 (ratio = 0.9), which was comparable to the stripe-necked turtle's performance. However, the red-eared slider exhibited a moderate decrease in performance in high ratio tests, indicating a potentially enhanced cognitive capacity to adapt to novel challenges. Our experimental design is repeatable and is adaptable to a range of freshwater turtles. These findings emphasize the potential importance of cognitive research to the underlying mechanisms of successful species invasions.

Geographic variation in incubation temperatures promoting viable offspring production in broadly co-distributed turtles.

Organisms whose early life stages are environmentally sensitive produce offspring within a relatively narrow range of suitable abiotic conditions. In reptiles, development rate and survival are often maximized if incubation temperatures remain under 31°C, though this upper bound may vary within and among species. We addressed this expectation by comparing responses to egg incubation at 30°C versus 33°C in congeneric turtle species pairs with broad syntopic geographic distributions. In the two softshell turtles (Apalone spp.), the greatest changes in development rate and phenotypic variance were observed in the northernmost population, which had a low survival rate (40%) at 33°C. The presumably suboptimal temperature (33°C) for northern populations otherwise yielded 76%-93% survival rates and fast swimming speeds in more southern populations. Still, in one species, northern hatchlings incubated at 33°C matched the elevated speeds of their southern counterparts, revealing a countergradient response. In northern populations of the two map turtles (Graptemys spp.), survival was also reduced (28%-60%) at 33°C and the development rate (relative to 30°C) increased by up to 75%. Our experiments on divergent taxa with similar nesting ecologies substantiate that the optimal thermal range for offspring production is variable. These findings encourage further work on how population- and species-level differences relate to local adaptation in widely distributed oviparous species.

Influence of incubation temperature, maternal effects, and paternity on quality of olive ridley hatchlings (Lepidochelys olivacea) from a mass-nesting beach in the Mexican Pacific.

Future climate change scenarios project that the increase in surface temperatures will affect ocean temperatures, inducing shifts in marine biodiversity. Sea turtles are species that are particularly vulnerable to the effects of climate change because temperature is a factor that influences embryonic development. We collected clutches of olive ridley turtles from a mass-nesting beach in the Mexican Pacific, which were incubated in ex situ conditions. When the hatchlings emerged, we measured the body condition index-which evaluates the weight-length relationship-and swim thrust, both were considered traits associated with fitness, termed "fitness proxies," and evaluated the effects of incubation temperature, maternal effects, and paternity on these fitness proxies. The body condition index was correlated positively and significantly with the arribada month and temperature during the last third of the incubation period but showed an inverse relationship with the maternal effect. While swim thrust was positively correlated with the maternal effect and the arribada month, there was an inverse relationship with incubation temperature during the first third of the period. Paternity, whether single or multiple, did not have a significant effect on either fitness proxies; however, it may have effects on the average fitness of a population of hatchlings. These results underscore the need to expand research on the sublethal effects of high incubation temperatures on the adaptation and survival of sea turtles, particularly in scenarios of rapid climate change.

An 18-µm microaggregate blood filter does not cause hemolysis during in vitro whole blood transfusions in sea turtles.

OBJECTIVE: Determine the hemolytic effect of an 18-µm microaggregate blood filter during in vitro sea turtle whole blood transfusions as well as describe the average diameter of leatherback (Dermochelys coriacea) and Kemp's ridley sea turtle (Lepidochelys kempii) RBCs. ANIMALS: 5 green (Chelonia mydas), 5 loggerhead (Caretta caretta), and 5 Kemp's ridley sea turtles (total n = 15). METHODS: Heparinized sea turtle blood was infused at 60 mL/h through a microbore extension set without and then with a postsyringe, inline 18-µm microaggregate blood filter. Pre- and postfiltration PCV, Hct, total solids, sodium, chloride, potassium, glucose, and free plasma hemoglobin concentrations were measured. With the use of light microscopy and archived blood smears, the maximum and minimum diameter of 20 RBCs from each of the 5 leatherback and 5 Kemp's ridley sea turtles were measured with a calibrated ocular micrometer using 400X magnification. RESULTS: There were no significant differences between pre- and postfiltration samples for Hct, total solids, sodium, chloride, potassium, glucose, and free plasma hemoglobin concentrations; however, there was a significant median postfiltration decrease in PCV of approximately 4%, representing a 13% decrease of the total RBCs transfused. Average maximum diameters for leatherback and Kemp's ridley sea turtle RBCs were 19.7 and 16.1 µm, respectively. CLINICAL RELEVANCE: Although the 18-µm microaggregate blood filter does not hemolyze transfused sea turtle RBCs and is likely safe for in vivo blood transfusions, the filter's pores may retain a small proportion of infused RBCs given their diameter.

Husbandry Protocols for Juvenile Loggerhead Sea Turtles (Caretta caretta) Based on Stress Response to Stocking Density and Dry-Dock Time.

When necessary, sea turtles are held captive for veterinarian care and research purposes. Protocols and basic guidelines have been described for husbandry of sea turtles with veterinarian needs but not considering physiological indicators of animal welfare. Because all sea turtle are imperiled species, monitoring their welfare is important. The aim of this study was to standardize husbandry protocols for loggerhead (Caretta caretta) juveniles held under seminatural conditions, based on circulating concentration of plasma corticosterone (Cort) and behavior. Two experiments were performed to analyze physiological and behavioral responses of the animals facing changes in stocking density and different dry-docking times. Cort analyses suggested that the number of animals per tank can be modified occasionally, without affecting their health and welfare. However, dry-docking time should be < 30 min, as indicated by the significant elevation of circulating Cort at ≥ 30 min, rising from 1.51- ng/ml to 5.28-ng/ml. Protocols tested did not affect behavioral responses, except for the breaths per move, which increased while Cort increased, despite differences exhibited by experimental animals in behavioral responses according to daily times (morning vs afternoon) and the sex of the animals.

Establishment and characterization of turtle liver organoids provides a potential model to decode their unique adaptations.

Painted turtles are remarkable for their freeze tolerance and supercooling ability along with their associated resilience to hypoxia/anoxia and oxidative stress, rendering them an ideal biomedical model for hypoxia-induced injuries (including strokes), tissue cooling during surgeries, and organ cryopreservation. Yet, such research is hindered by their seasonal reproduction and slow maturation. Here we developed and characterized adult stem cell-derived turtle liver organoids (3D self-assembled in vitro structures) from painted, snapping, and spiny softshell turtles spanning ~175My of evolution, with a subset cryopreserved. This development is, to the best of our knowledge, a first for this vertebrate Order, and complements the only other non-avian reptile organoids from snake venom glands. Preliminary characterization, including morphological, transcriptomic, and proteomic analyses, revealed organoids enriched in cholangiocytes. Deriving organoids from distant turtles and life stages demonstrates that our techniques are broadly applicable to chelonians, permitting the development of functional genomic tools currently lacking in herpetological research. Such platform could potentially support studies including genome-to-phenome mapping, gene function, genome architecture, and adaptive responses to climate change, with implications for ecological, evolutionary, and biomedical research.

Experimental study on color and texture as cues for plastic debris ingestion by captive sea turtles.

Sea turtles face considerable risks from ingesting marine debris. They are primarily visual feeders, so color may be important for identifying food suitability or enhancing prey detection. Here, we investigated the impacts of color and texture on foraging behavior in relation to plastic consumption. We experimentally assessed the influences of color and texture as attractors for sea turtles using edible jellyfish. The findings showed that the colors of objects significantly affected selective preferences, as evidenced by different behaviors by sea turtles in response to different colors. They exhibited diet-related selectivity toward colors similar to common aquarium food, and texture had a significant impact on complete ingestion. The results suggest that plastic resembling natural prey is more likely ingested. Also, sea turtles were attracted by the color yellow, suggesting that visually distinctive items can attract them. Our results provide fundamental knowledge, helping mitigate the effects of plastic pollution on wildlife.

Anthropogenic noise predicts sea turtle behavioural responses.

Anthropogenic noise is a pollutant of global concern. While the effects of underwater noise pollution have been frequently studied in fish and mammals, our understanding of how this anthropogenic stressor affects marine reptiles is scant. Using a multichannel data logger equipped with a camera and hydrophone, we quantified behavioural responses of a free-ranging green turtle (Chelonia mydas) to vessel noise in the Galapagos Archipelago, an important nesting site in the eastern Pacific. We found that while travelling the turtle increased its vigilance with increasing vessel noise. However, when on the seabed the turtle did not increase its vigilance with increasing noise levels. Our findings illustrate that noise pollution has the potential to alter overall time budgets of animals. Identifying real-time responses of wild animals illustrate how in situ approaches allow to assess the effects of human activities on marine systems.

Adaptation of sea turtles to climate warming: Will phenological responses be sufficient to counteract changes in reproductive output?

Sea turtles are vulnerable to climate change since their reproductive output is influenced by incubating temperatures, with warmer temperatures causing lower hatching success and increased feminization of embryos. Their ability to cope with projected increases in ambient temperatures will depend on their capacity to adapt to shifts in climatic regimes. Here, we assessed the extent to which phenological shifts could mitigate impacts from increases in ambient temperatures (from 1.5 to 3°C in air temperatures and from 1.4 to 2.3°C in sea surface temperatures by 2100 at our sites) on four species of sea turtles, under a "middle of the road" scenario (SSP2-4.5). Sand temperatures at sea turtle nesting sites are projected to increase from 0.58 to 4.17°C by 2100 and expected shifts in nesting of 26-43 days earlier will not be sufficient to maintain current incubation temperatures at 7 (29%) of our sites, hatching success rates at 10 (42%) of our sites, with current trends in hatchling sex ratio being able to be maintained at half of the sites. We also calculated the phenological shifts that would be required (both backward for an earlier shift in nesting and forward for a later shift) to keep up with present-day incubation temperatures, hatching success rates, and sex ratios. The required shifts backward in nesting for incubation temperatures ranged from -20 to -191 days, whereas the required shifts forward ranged from +54 to +180 days. However, for half of the sites, no matter the shift the median incubation temperature will always be warmer than the 75th percentile of current ranges. Given that phenological shifts will not be able to ameliorate predicted changes in temperature, hatching success and sex ratio at most sites, turtles may need to use other adaptive responses and/or there is the need to enhance sea turtle resilience to climate warming.

Morphological Analysis of the Olfactory System of the Pig-Nosed Turtle, Carettochelys insculpta.

The turtle olfactory organ consists of the upper (UCE) and lower (LCE) chamber epithelium, projecting to the ventral and dorsal parts of the olfactory bulbs, respectively. The UCE is associated with glands, contains ciliated olfactory receptor neurons, and is assumed to detect odorants primarily in air, while the LCE is devoid of glands, contains microvillous olfactory receptor neurons, and is assumed to detect odorants primarily in water. Examining the olfactory system of the pig-nosed turtle, Carettochelys insculpta, this study found that both the upper and lower chambers of the nasal cavity were lined with sensory epithelium devoid of associated glands and contained ciliated olfactory receptor neurons. Moreover, the olfactory bulbs were not divided into dorsal and ventral parts. These results suggest that the olfactory system of the pig-nosed turtle is a single system specialized for detecting odorants in water.

Daily, seasonal, and long-distance movements inferred from Fastloc-GPS telemetry of immature green turtles (Chelonia mydas) at a high-latitude, mid-ocean developmental site.

To characterize the movements and habitat use of juvenile green turtles (Chelonia mydas) in benthic developmental habitat, we deployed Fastloc-GPS-enabled satellite transmitters on 16 individuals captured as part of a multi-decade study of green turtles on the Bermuda Platform. We characterized residence areas, distinct use areas within them, and seasonal movements based on an average of 562 Fastloc-GPS positions and 284 tracking days per turtle. We estimated residence area sizes using traditional home range methods, e.g., 90% utilization distribution (UD) (mean 2.29 ±2.71 km2) and 50% UD (mean 0.54 ±0.69 km2). Total residence area size increased significantly over the 8-year study, from <1 km2 before 2013 to ≥3 km2 in 2018 (R2 = 0.51, F1,14 = 14.55, p = 0.0019), corresponding to a period of decline in seagrass habitat and suggesting increased foraging effort. We identified three types of distinct use areas within residence areas where tracked turtles typically exhibited behavioral fidelity: foraging, resting, and cool weather refugia. These distinct use areas were smaller than high-use areas from previous studies; e.g., seagrass meadow foraging areas averaged 0.05 km2. Most turtles made daily transits between foraging and resting sites; for some individuals, these involved crossing frequently used vessel navigation channels. Seasonal variation in behavior suggested that the overwintering strategy for green turtles on the Bermuda Platform involves "optional dormancy," during which turtles spent less time on seagrass meadows and made brief excursions to distinct deeper habitats. Four individuals made directed (mean path straightness = 0.93 ±0.02 SD) developmental migrations away from Bermuda toward known adult foraging range. Results of our study further knowledge of the green turtle life cycle at a high-latitude site; they demonstrate that green turtles show fidelity to distinct use areas within developmental habitats over many years and exhibit seasonal movements.

Effects of temperature heterogeneity on freshwater turtle habitat selection at their northern range limit.

Environmental temperature is a crucial resource for ectotherms, affecting their physiology, behaviour and fitness. To maintain body temperatures within a suitable performance range, ectotherms select thermally-favourable locations, but this selection may be challenging in environments with high spatio-temporal heterogeneity. We assessed thermal habitat selection in two freshwater turtles (Emydoidea blandingii; Chrysemys picta) within a thermally heterogeneous environment at two spatial scales (selection of home ranges within the landscape, selection of locations within home ranges) and across seasons, by comparing temperatures at turtle locations vs. those available in the environment. Turtles selected warmer locations compared to those available in aquatic and terrestrial habitats only within home ranges, but did not show any temperature preferences when selecting home ranges at the larger scale. Turtles selected locations that were less thermally-variable than their surroundings, both at the home range scale and within home ranges. Thermal habitat selection was strongest during colder and more thermally-variable pre-nesting season compared to later periods. Despite differences in thermal mass between species, both species responded similarly to temperature variation. We conclude that freshwater turtles at their northern range margin select suitable microclimates within the suite of conditions that are naturally available.

Incubation in shaded hatcheries biases sex-determination but preserves Lepidochelys olivacea hatchling physiology.

Some studies have associated ex situ conservation with cerebral and gonadal developmental delay, as well as decreased motor performance in Lepidochelys olivacea offspring. Ex situ management is also related to a more mature spleen and a differential leukocyte count in newly emerged Lepidochelys olivacea hatchlings. The physiological relevance of a more mature spleen is unknown in sea turtles, but studies in birds suggest an increased immune response. Because egg relocation to hatcheries is a common conservation practice, it is imperative to know its impact on hatchling physiology. Herein, plasma activity of superoxide dismutase, alkaline phosphatase and the alternative complement pathway, as well as total antioxidant capacity and hydrogen peroxide concentrations were quantified in hatchlings from in situ and ex situ nests under basal conditions at nest emergence. Toll-like receptor 4 (tlr4), heat shock proteins (hsp) 70 and hsp90 expression were quantified in the spleen and liver of the hatchlings. Hepatocyte density and nuclear area were quantified in histological sections of the liver and all turtles were sexed by histological sectioning of the gonads. Total antioxidant capacity and hydrogen peroxide concentrations in plasma were lower in turtles from ex situ nests, while tlr4 and hsp70 mRNA expression was higher in the spleen but not in the liver. Ex situ incubation produced 98% male hatchlings, whereas in situ incubation produced 100% females. There were no other differences in the attributes sampled between hatchlings emerging from ex situ and in situ treatments. The results suggest that ex situ relocated turtles may be less prone to oxidative stress than in situ incubated hatchlings and could have more mature splenic function. Together, the data suggest that ex situ relocation to shaded hatcheries biased sex determination but preserved the general physiological condition of sea turtle hatchlings.