Temperature-dependent Sex Determination Research Articles

Re-Equilibrating Sex Ratios: Adjustment of Reaction Norms in Species With Temperature-Dependent Sex Determination.

Fisher's general principle for sex allocation holds that population sex ratios are typically balanced because parents producing the rare sex are benefited and the rare sex alternates over time. In species that have temperature-dependent sex determination (TSD), thermal reaction norms need to be adjusted at the population level to avoid extremely biased sex ratios and extinction. Extant species with TSD experienced drastic climatic changes in the geological past and must necessarily have mechanisms of adaptation. I propose here a conceptual framework to explain how TSD curves could be adjusted by means of natural selection, based on Fisher's equilibrium sex-ratio principle. Through a process that alternatively favors mothers that tend to produce the rare sex under new temperatures, sex ratios eventually return toward a theoretical equilibrium. Prerequisites for this model are variability among mothers in the tendency to produce a particular sex at a given temperature (i.e., variability in the thermal reaction norm), inheritance of this trend, and higher fitness of the rare sex. This straightforward mechanism could facilitate thermal adaptation in species with TSD over multiple generations.

Adult Sex-Ratio Bias Does Not Lead to Detectable Adaptive Offspring Sex Allocation Via Nest-Site Choice in a Turtle With Temperature-Dependent Sex Determination.

Sex-ratio theory predicts that parents can optimise their fitness by producing offspring of the rare sex, yet there is a dearth of empirical evidence for adaptive sex allocation in response to the adult sex ratio (ASR). This is concerning, as anthropogenic disruption of the sex ratios of reproductive individuals threatens to cause demographic collapse in animal populations. Species with environmental sex determination (ESD) are especially at risk but may possess the capacity to adaptively influence offspring sex via control over the developmental environment. For example, reptiles with temperature-dependent sex determination (TSD) could conceivably choose nest sites with thermal characteristics that produce offspring of the rare sex. To test this hypothesis, we seeded three secure outdoor ponds with different sex ratios (~M:F 3:1, 1:1, and 1:3) of adult painted turtles (Chrysemys picta), a reptile species with TSD. We then quantified nesting traits that could influence nest temperature and thus offspring sex ratio, including nesting date, nest depth, and nest canopy cover. We found no directional relationship between the ASR treatments and any measured nest traits and thus rejected our hypothesis. Interestingly, increased maternal body size was associated with reduced nest canopy cover, and this trend was more pronounced in the biased ASR treatments. If adaptive sex allocation occurs in this system, it instead may manifest via maternal epigenetic predisposition of offspring sex or in response to a phenomenon other than the ASR.

Developmental Thermal Reaction Norms of Leatherback Marine Turtles at Nesting Beaches.

Accurate scientific information is critical for undertaking appropriate conservation and management practices for imperiled species. One source of concern is that research findings might vary for non-biological reasons, including experimental design and analytical methods. To illustrate, we provide detailed modern analysis of reproductive data for leatherback turtles (Dermochelys coriacea). This species exhibits significant fluctuations in nesting densities across different regions, possibly driven by local rather than global factors. Key factors influencing these changes include hatching success and sex determination, both sensitive to incubation temperatures (e.g., lower temperatures yield more males, higher temperatures yield females). This study updates the understanding of temperature-dependent sex determination (TSD) in this species using Bayesian statistics. Growth rate data from the West Pacific and Northwest Atlantic populations show a similar, monotone increase with temperature, affirming the reliability of the models used. The analysis of TSD patterns indicates that observed differences are more likely due to study methodologies and clutch-specific factors rather than regional differences. These findings challenge previous assumptions, showing that leatherback TSD does not conform to a simple on/off pattern but is influenced by multiple, interacting environmental factors. Population dynamics models must account for these complexities, recognizing that both sex ratios and hatching success are critical to understand the rapid changes observed in some leatherback populations.

The extraordinary Atlantic silverside and me

Abstract My career is inextricably linked to the remarkable characteristics and astonishing insights provided by a simple yet elegant species, the Atlantic silverside, Menidia menidia. My initial field and laboratory studies provided the first demonstration of temperature-dependent sex determination (TSD) and its adaptive significance in fish, as well as the first empirical confirmation of frequency-dependent sex ratio selection. This early work then led to the discovery of countergradient variation in growth, evolutionary trade-offs associated with growth, pioneered the study of fisheries-induced evolution, and furthered our understanding of the spatial scale of local adaptation in marine species. Most of these topics are now vigorous areas of research encompassing numerous species and systems. Here I describe how these unforeseen discoveries were made and led to other findings on bluefish (Pomatomus saltatrix) and striped bass (Morone saxatilis). An interest in leadership during my early years was later rekindled as I took on administrative roles. It all began with an intriguing observation stemming from my MS thesis that I was compelled to follow. My career illustrates the value of exploring deeply the natural history of lesser-known yet easy-to-study species and to follow unexpected observations even if peripheral to a preconceived plan of research.

PSTAT3 activation of Foxl2 initiates the female pathway underlying temperature-dependent sex determination

Ovarian development was traditionally recognized as a "default" sexual outcome and therefore received much less scientific attention than testis development. In turtles with temperature-dependent sex determination (TSD), how the female pathway is initiated to induce ovary development remains unknown. In this study, we have found that phosphorylation of the signal transducer and activator of transcription 3 (pSTAT3) and Foxl2 exhibit temperature-dependent sexually dimorphic patterns and tempo-spatial coexpression in early embryos of the red-eared slider turtle (Trachemys scripta elegans). Inhibition of pSTAT3 at a female-producing temperature of 31 °C induces 64.7% female-to-male sex reversal, whereas activation of pSTAT3 at a male-producing temperature of 26 °C triggers 75.6% male-to-female sex reversal. In addition, pSTAT3 directly binds to the locus of the female sex-determining gene Foxl2 and promotes Foxl2 transcription. Overexpression or knockdown of Foxl2 can rescue the sex reversal induced by inhibition or activation of pSTAT3. This study has established a direct genetic link between warm temperature-induced STAT3 phosphorylation and female pathway initiation in a TSD system, highlighting the critical role of pSTAT3 in the cross talk between female and male pathways.

Empirical evidence for the extent of spatial and temporal thermal variation on sea turtle nesting beaches

Recording sand temperatures has become routine at many sea turtle nesting sites across the world given the impacts of incubation temperatures on hatchling sex ratios. However, the extent of thermal variability found at a nesting site has previously received little attention. Here we examine empirical sand temperature records across five atolls extending 250 km in the Chagos archipelago, Indian Ocean, between October 2012 and July 2023 and quantify the extent of spatial and temporal thermal variability. Our results suggest that sand temperatures at our study site vary seasonally and inter-annually, between beaches in the archipelago, and within beaches in different nesting habitats. The biggest drivers of thermal variability were seasonal and inter-annual differences, which modulated sand temperatures by up to 3.00°C and 1.03°C, respectively. Intra-beach and inter-beach variability further modulated temperatures by up to 0.56°C and 0.85°C, respectively. In addition, mean monthly sand temperatures were relatively low, suggesting that hatchling sex ratios are fairly balanced. The wide range of sand temperatures recorded at this nesting site suggests that it is likely both male-biased and female-biased clutches are produced during the nesting season. Quantifying thermal variability from a long-term sand temperature time series offers valuable insight into a population with temperature-dependent sex determination and, when possible, should be considered when modelling temperature impacts on hatchling sex ratios.

Temperature-Dependent Sex Determination in Crocodilians and Climate Challenges.

The sex of crocodilians is determined by the temperature to which the eggs, and hence the developing embryo are exposed during critical periods of development. Temperature-dependent sex determination is a process that occurs in all crocodilians and numerous other reptile taxa. The study of artificial incubation temperatures in different species of crocodiles and alligators has determined the specific temperature ranges that result in altered sex ratios. It has also revealed the precise temperature thresholds at which an equal number of males and females are generated, as well as the specific developmental period during which the sex of the hatchlings may be shifted. This review will examine the molecular basis of the sex-determination mechanism in crocodilians elucidated during recent decades. It will focus on the many patterns and theories associated with this process. Additionally, we will examine the consequences that arise after hatching due to changes in incubation temperatures, as well as the potential benefits and dangers of a changing climate for crocodilians who display sex determination based on temperature.

The thermal sensitivity of growth and survival in a wild reptile with temperature‐dependent sex determination

The Charnov‐Bull hypothesis is the leading explanation for the evolution of environmental sex determination (ESD), which includes temperature‐dependent sex determination (TSD), the most common form of ESD. Charnov‐Bull predicts a sex‐by‐incubation temperature interaction for fitness, matching offspring sex with thermal conditions that increase parental fitness. However, there is no general explanation for how the sex‐by‐temperature interaction arises. Two competing explanations for the interaction lie in the survival to maturity hypothesis (SM) and the Trivers–Willard extension (TW). Under SM, the sex that matures later is produced under optimal incubation regimes as the late‐maturing sex accrues more mortality by maturation, while TW suggests that males are always produced under optimal incubation regimes as male mating success is more sensitive to condition (general health, vigor) than female fecundity. In a system where females mature later than males, as in the painted turtle Chrysemys picta, SM and TW generate opposite predictions for the effect of incubation temperature on juvenile survival. We incubated C. picta eggs under either female‐promoting temperatures (28 ± 3 °C) or male‐promoting temperatures (25 ± 3 °C), then released the hatchlings into their natal pond. We used a Bayesian capture–mark–recapture approach to follow their survival over two growing seasons. We found a 2% depression of biweekly survival in individuals incubated under the cooler temperature, providing subtle support for SM. Incubation treatments did not influence growth. Large‐scale field experiments such as this one will be necessary for understanding TSD evolution, and we underline general principles to execute such experiments successfully.

Gene expression dynamics during temperature-dependent sex determination in a sea turtle

Fifty years ago, researchers discovered a link between ambient temperature and the sex of turtle embryos. More recently, significant progress has been made in understanding the influence of temperature on freshwater turtles. However, our understanding of the key genetic factors in other turtle groups, such as sea turtles, remains limited. To address this gap, we conducted RNA-seq analyses on embryonic tissues from the sea olive ridley turtle during the thermosensitive period (stages 21–26) at temperatures known to produce males (26 °C) and females (33 °C). Our findings revealed that incubation temperatures primarily influence genes with broad expression across tissues due to differential cell division rates and later have an effect regulating gonad-specific transcripts. This effect is mostly related to gene activation rather than transcription repression. We performed transcriptome analyses following shifts in incubation temperatures of bi-potential gonads. This approach allowed us to identify genes that respond rapidly and may be closer to the beginning of the temperature-sensing pathway. Notably, we observed swift adaptations in the expression levels of chromatin modifiers JARID2 and KDM6B, as well as the splicing factor SRSF5, and transcription regulators THOC2, DDX3X and CBX3, but little impact in the overall gonad-specific pathways, indicating that temperature-sensing genes may change rapidly but the rewiring of the gonad's developmental fate is complex and resilient. Author summarySea turtles, one of the most iconic creatures of our oceans, confront a troubling reality of endangerment, a peril magnified by the looming specter of climate change. This climatic shift is gradually increasing the temperature of the nesting beaches thus causing dramatic male/female population biases. Conservation efforts will need genetic and molecular information to reverse the negative effects of climate change on the populations. In this study, we conducted the first transcriptomic analysis of embryonic tissues, including gonads, brain, liver, and mesonephros, in the olive ridley sea turtle during the critical thermosensitive period spanning stages 21–26. We examined both male-producing (26 °C) and female-producing (33 °C) temperatures and found that incubation temperatures influence temperature-sensitive genes that are either expressed globally or specifically associated with the gonads. These findings indicate that incubation temperatures predominantly sway genes with broad expression patterns due to differential cell division rates. This natural process was opted in the gonads to drive sex determination. We also identified genes that are rapidly capable of sensing temperature changes and that could play a role in the activation of the sex determination pathway. Overall, our study sheds light on the intricate interplay between temperature and gene expression during sea turtle development, revealing dynamic changes in the transcriptome and highlighting the involvement of key genetic players in sex determination.

Temperature regulates sex determination and growth in the paralichthid flatfishCalifornia halibut.

California halibut (Paralichthys californicus) is a candidate species for aquaculture and stock enhancement. These applications rely on sex control, either to maximize the production of faster growing females or to match sex ratios in the wild. Other paralichthids exhibit temperature-dependent sex determination (TSD), but the presence and pattern of TSD is not well defined in California halibut. Juvenile California halibut were cultured at three distinct temperatures (15°C, 19°C, and 23°C) through the developmental period presumed to be thermosensitive based on findings from congeners. Sex ratios were quantified in each treatment using phenotypic sex identification techniques applied early (molecular biomarkers; 51-100 mm total length [TL]) and late (visual examination of the gonads; ≥100 mm TL) in the juvenile phase. Both techniques indicated similar sex determination trends at each temperature, with overall sex ratios assessed as 49.9% maleat 15°C, 74.5% maleat 19°C, and 98.2% maleat 23°C. Growth rates were highest at 23°C and lowest at 15°C, with intrinsically fast- and slow-growing individuals at all temperatures. At 15°C and 19°C, females comprised a higher proportion among the fast growers than they did among the slow growers. These data show that California halibut exhibit TSD, with temperatures of 19°C and 23°C masculinizing fish while 15°C appears to produce a 1:1 sex ratio. This study will help optimize sex ratios and growth in hatcheries through thermal manipulation. Furthermore, the developed biomolecular tools and identified temperature thresholds will be important in future work to understand the influence of global warming on wild population demographics.

Screening of temperature-responsive signalling molecules during sex differentiation in Asian yellow pond turtle (Mauremys mutica)

BackgroundThe Asian yellow pond turtle (Mauremys mutica) is an important commercial freshwater aquaculture species in China. This species is a highly sexually dimorphic species, with males growing at a faster rate than females and exhibits temperature-dependent sex determination (TSD), in which the incubation temperature during embryonic development determines the sexual fate. However, the mechanisms of the sex determination or sex differentiation in the Asian yellow pond turtle are remain a mystery.ResultsTemperature-specific gonadal transcriptomics of the Asian yellow pond turtle were performed during the thermosensitive period (stage 15) using RNA-seq technology to identify candidate genes that initiate gonadal differentiation. We uncovered candidates that were the first to respond to temperature. These candidates were sexually dimorphic in expression, reflecting differences in gonadal (Cirbp, Runx1) and germline differentiation (Vasa, Nanos1, Piwil2), gametogenesis (Hmgb3, Zar1, Ovoinhibitor-like, Kif4), steroid hormone biosynthesis (Hsd17b5, Hsd17b6), heat shock (Dnajb6, Hsp90b1, Hsp90aa1) and transient receptor potential channel genes (Trpm1, Trpm4, Trpm6, Trpv1).ConclusionsOur work will provide important genetic information to elucidate the mechanisms of sex control in the Asian yellow pond turtles, and will contribute important genetic resources for further studies of temperature-dependent sex determination in turtles.

Environmental (and Random?) Sex Determination in Endangered and Invasive Phelsuma Geckos

Introduction: Sex is a fundamental characteristic of an individual. It is therefore puzzling why in some systems sex is precisely determined by a genotype, while in others it is influenced by the environment or even subtle, not to say random, factors. Some stochasticity in sex determination would be expected if environmental conditions did not have a large sex-specific effect on fitness. Although data are only available for a small fraction of species, geckos show exceptional variability in sex determination. Methods: We tested the effects of three incubation temperatures on sex ratio and adult body size in the invasive gecko Phelsuma laticauda and the vulnerable gecko Phelsuma nigristriata. Results: We document temperature-dependent sex determination (TSD) in both species. Only females hatched at a low temperature (24°C), whereas male production peaked at an intermediate temperature (28°C) and declined, at least in P. laticauda, again at the highest temperature (31°C). Interestingly, full siblings hatched from eggs glued together during the incubation at temperatures producing both sexes are often of the opposite sex. We found no significant effect of incubation temperature on adult body length. Conclusions: Documentation of TSD in the day geckos has implications for conservation practice in environmental management of endangered species or eradication of invasive species. However, it appears that a very subtle (random?) factor may also be involved in their sex determination. In line with this, we found no significant effect of incubation temperature on adult body length.

Reconstructed and projected beach temperatures reveal where flatback turtles are most at risk from climate change

Nest temperature is the predominant driver of emergence success and primary sex ratios in sea turtles, with female offspring produced at higher temperatures due to temperature-dependent sex determination. However, emergence success and primary sex ratios are unfeasible to measure at scale, making methods to estimate these life-history traits from predicted sand temperatures highly desirable for long-term conservation planning in the context of climate change. To address this, we used a mechanistic microclimate model to predict hourly sand temperatures, over 32 nesting seasons since 1986, at 402 West Australian beaches supporting nesting by flatback turtles (Natator depressus). Predicted sand temperatures indicated that ∼70% of these beaches carried a 'very low' to 'intermediate' risk of subjecting embryos to thermal stress. By combining these temperature predictions with a physiological model, current and future emergence success and sex ratios were projected for ten different beaches spanning a range of thermal microclimates, under various climate change scenarios. Under recent climate conditions, emergence success averaged 76%, but declined to 63% and 37% with a 2°C and 4°C increase in air temperature, respectively. The sex ratios of hatchlings varied by location, but extremely skewed sex ratios were anticipated in a 4°C warming scenario. Our projections reveal that 'high risk' nesting beaches will regularly experience clutch failure as climate change progresses, while cooler beaches offer long-term nesting potential and require protection from additional anthropogenic impacts. These projections, covering an entire genetic stock, supply demographic data for assessing extinction risks and this method can be applied to sea turtle populations worldwide.

Assessment of Acetylcholinesterase Enzyme (AChE) Activity in the Developing Brain of TSD Lizard, Calotes Versicolor (Daud.)

Acetylcholinesterase (AChE) is a critical enzyme in the neuronal cholinergic system of vertebrates. It is one of the efficient cholinesterases that is involved in the termination of acetylcholine-mediated neurotransmission, rapidly hydrolyzing acetylcholine into acetate and choline. Barring a few reports on the distribution/presence of AChE enzyme activity in the adult reptilian species there are nil reports on the presence of this enzyme activity in the developing brain. Hence, the present investigation aims to evaluate AChE enzyme activity in the developing brain of an oviparous lizard, Calotes versicolor which exhibits a novel Female-Male-Female-Male (FMFM) pattern of temperature-dependent sex determination (TSD). The eggs were collected during breeding season, padded with moist cotton, and incubated in an incubator at 30o C. The whole brain from the embryos was collected during different developmental stages from oviposition (Stage 27) to hatching (stage 42). The AChE enzyme activity was quantified in the brain, according to Ellman’s protocol using a spectrophotometer. The findings reveal that the onset of AChE enzyme activity is observed as early as at oviposition, which reflects the early action of AChE activity in the developing brain. The observed results albeit indirectly suggest the involvement of AChE enzyme activity in the morphogenetic process. Further, an exponential increase in AChE activity during the post-gonadal differentiation phase indicates age-related elevation in AChE enzyme activity which in turn unveils its involvement in neuronal transmission of the embryonic brain. Besides, this specific AChE enzyme activity in the developing brain of Calotes versicolor represents a conserved pattern for the cholinergic system in vertebrates.

Mathematical modelling to assess the impact of stress on temperature-dependent sex determination in teleost fish

Mathematical modelling to assess the impact of stress on temperature-dependent sex determination in teleost fish

Overexpression of Kdm6b induces testicular differentiation in a temperature-dependent sex determination system.

In reptiles, such as the red-eared slider turtle ( Trachemys scripta elegans), gonadal sex determination is highly dependent on the environmental temperature during embryonic stages. This complex process, which leads to differentiation into either testes or ovaries, is governed by the finely tuned expression of upstream genes, notably the testis-promoting gene Dmrt1 and the ovary-promoting gene Foxl2. Recent studies have identified epigenetic regulation as a crucial factor in testis development, with the H3K27me3 demethylase KDM6B being essential for Dmrt1 expression in T. s. elegans. However, whether KDM6B alone can induce testicular differentiation remains unclear. In this study, we found that overexpression of Kdm6b in T. s. elegans embryos induced the male development pathway, accompanied by a rapid increase in the gonadal expression of Dmrt1 at 31°C, a temperature typically resulting in female development. Notably, this sex reversal could be entirely rescued by Dmrt1 knockdown. These findings demonstrate that Kdm6b is sufficient for commitment to the male pathway, underscoring its role as a critical epigenetic regulator in the sex determination of the red-eared slider turtle.

Sex Ratio Estimation of Green Turtle Hatchlings in Kazanlı Beach, Türkiye

Sea turtles are distributed worldwide, mostly in the tropical and subtropical waters. All seven species of sea turtles in the world have temperature-dependent sex determination. The pivotal temperature for hatchlings with a female:male ratio of 1:1 is usually around 29°C. Female hatchlings are observed at values higher than the pivotal temperature and male hatchlings at lower values. It is expected that wildlife threatened by global warming will be more severely affected, especially species that sex is determined by temperature. This study was carried out by measuring air, sand and nest temperatures during two nesting seasons at Kazanlı Beach, one of the largest nesting sites of Chelonia mydas (Green turtle) in the Mediterranean. Sand grain size was also measured, and gonads of dead hatchlings were examined histologically. Both sand (n=4) and nest (n=6) temperatures were above the pivotal temperature (between 29.1-31.6°C) in the study area. According to the estimation based on temperatures, hatchlings were female (♀>50%) biased. Of the hatchling samples, 75 were examined histologically. According to histological examination, the female:male ratio was 2:1. According to the direct examination and indirect estimation; the sex ratio of green turtle hatchlings in Kazanlı Beach, one of the highest hatchling producing nesting sites of the Mediterranean, was reported for the first time in this study. There is a need to fill the data gaps regarding sex ratios, especially for major beaches with high hatchling production. Knowing the temperature profiles in and around the nest and sex ratios of hatchlings is critical for the development of conservation measures for endangered sea turtles.

Feminization of a mixed-stock foraging aggregation of immature green turtles (Chelonia mydas), 1975–2018

Marine turtles exhibit temperature-dependent sex determination (TSD), and there is widespread concern that global warming is raising nest incubation temperatures, resulting in increasingly female-skewed sex ratios in “feminized” populations. We assessed the sex ratio of a mixed-stock aggregation of immature green turtles (Chelonia mydas) at a midocean developmental foraging ground in the Northwest Atlantic from 1975 to 2018. We used plasma testosterone concentrations, measured by radioimmunoassay (RIA) and calibrated by laparoscopy, to determine the sex of 2,724 green turtles captured 3,940 times in Bermuda (32°18′ N, 64°46′ W) waters. A logistic regression model correctly predicted the sex of 99.5% of turtles (189/190) with associated testosterone concentrations whose sex had been verified via laparoscopy. Empirical evaluation of the trend in sex ratio using four related data sets showed a significant increase (2.8–4.0% yr−1) in the sex ratio (F/M) over the course of the study. Using growth rates to predict the year of arrival of turtles in Bermuda, we estimated the sex ratio of recruiting cohorts over 4 decades. Mixed-stock analysis of mtDNA sequences of 602 turtles that recruited to Bermuda between 1970 and 2018 suggested that multiple, geographically dispersed rookeries contributed to the Bermuda aggregation making it regionally representative. Changes in rookery contributions and strong population increases at certain rookeries may partly explain the increasing trend in the sex ratio. But the steady rate of increase over decades and the increasing female percentage of arriving cohorts are consistent with impacts of global warming at source rookeries.

Exploring contaminants as a disruptor of temperature-dependent sex determination in sea turtle hatchlings

Sea turtle nesting beaches are experiencing increased sand temperatures as climate change progresses. In one major green turtle (Chelonia mydas) nesting beach in the northern Great Barrier Reef, over 99 percent of hatchlings are female. The effects of contaminants on sea turtle hatchling sex determination are not often explored. Liver samples were collected from green turtle hatchlings that were sacrificed for histological sex determination in a parallel study on the effects of sand cooling on sex ratios, and analysed for trace elements via acid digestion and organic contaminants via in vitro cytotoxicity bioassays. Chromium, antimony, barium, and cadmium have previously been demonstrated to be estrogenic, and concentrations of these elements were used to calculate three estrogenic indexes for each clutch: predicted relative estrogenic potency (PEEQA), the sum of percent trace elements above the median of all samples (TEOM), and the sum of percent estrogenic elements above the median of all samples (EstroEOM). Excluding an outlier clutch, cadmium, antimony, and EstroEOM had significant positive relationships with sex ratio deviation. Mean clutch cobalt, lead, antimony and barium, also had a significant positive relationship with clutch sex ratio. There was no relationship between in vitro cytotoxicity of liver extracts and sex ratio, however, 9% of hatchlings had organic contaminants high enough to suggest potential cellular damage. Contaminant effects on sex determination are likely to be caused by a mixture of contaminant interactions as well as temperature. Many trace elements detected in this study have also been linked to negative health effects on hatchlings in previous studies. Considering the risks of feminization due to climate change and potential contaminant effects on hatchling health and sex determination, future studies exploring contaminant effects on sea turtle hatchling sex determination are recommended.

Short-term resilience to climate-induced temperature increases for equatorial sea turtle populations.

Projection models are being increasingly used to manage threatened taxa by estimating their responses to climate change. Sea turtles are particularly susceptible to climate change as they have temperature-dependent sex determination and increased sand temperatures on nesting beaches could result in the 'feminisation' of hatchling sex ratios for some populations. This study modelled likely long-term trends in sand temperatures and hatchling sex ratios at an equatorial nesting site for endangered green turtles (Chelonia mydas) and critically endangered hawksbill turtles (Eretmochelys imbricata). A total of 1078 days of sand temperature data were collected from 28 logger deployments at nest depth between 2018 and 2022 in Papua New Guinea (PNG). Long-term trends in sand temperature were generated from a model using air temperature as an environmental proxy. The influence of rainfall and seasonal variation on sand temperature was also investigated. Between 1960 and 2019, we estimated that sand temperature increased by ~0.6°C and the average hatchling sex ratio was relatively balanced (46.2% female, SD = 10.7). No trends were observed in historical rainfall anomaliesand projections indicated no further changes to rainfall until 2100. Therefore, the sex ratio models were unlikely to be influenced by changing rainfall patterns. A relatively balanced sex ratio such as this is starkly different to the extremely female-skewed hatchling sex ratio (>99% female) reported for another Coral Sea nesting site, Raine Island (~850 km West). This PNG nesting site is likely rare in the global context, as it is less threatened by climate-induced feminisation. Although there is no current need for 'cooling' interventions, the mean projected sex ratios for 2020-2100 were estimated 76%-87% female, so future interventions may be required to increase male production. Our use of long-term sand temperature and rainfall trends has advanced our understanding of climate change impacts on sea turtles.