Chondrocranial anatomy of Testudo hermanni (Testudinidae, Testudines) with a comparison to other turtles.

Using histological cross-sections, the chondrocranium anatomy was reconstructed for two developmental stages of Hermann's tortoise (Testudo hermanni). The morphology differs from the chondrocrania of most other turtles by a process above the ectochoanal cartilage with Pelodiscus sinensis being the only other known species with such a structure. The anterior and posterior processes of the tectum synoticum are better developed than in most other turtles and an ascending process of the palatoquadrate is missing, which is otherwise only the case in pleurodiran turtles. The nasal region gets proportionally larger during development. We interpret the enlargement of the nasal capsules as an adaption to increase the surface area of the olfactory epithelium for better perception of volant odors. Elongation of the nasal capsules in trionychids, in contrast, is unlikely to be related to olfaction, while it is ambiguous in the case of Sternotherus odoratus. However, we have to conclude that research on chondrocranium anatomy is still at its beginning and more comprehensive detailed descriptions in relation to other parts of the anatomy are needed before providing broad-scale ecological and phylogenetic interpretations.

Identifying the source rookery of green turtles (Chelonia mydas) found in feeding grounds around the Korean Peninsula.

Determining the genetic diversity and source rookeries of sea turtles collected from feeding grounds can facilitate effective conservation initiatives. To ascertain the genetic composition and source rookery, we examined a partial sequence of the mitochondrial control region (CR, 796 bp) of 40 green turtles (Chelonia mydas) collected from feeding grounds around the Korean Peninsula between 2014 and 2022. We conducted genetic and mixed-stock analyses (MSA) and identified 10 CR haplotypes previously reported in Japanese populations. In the haplotype network, six, three, and one haplotype(s) grouped with the Japan, Indo-Pacific, and Central South Pacific clades, respectively. The primary rookeries of the green turtles were two distantly remote sites, Ogasawara (OGA) and Central Ryukyu Island (CRI), approximately 1,300 km apart from each other. Comparing three parameters (season, maturity, and specific feeding ground), we noted that OGA was mainly associated with summer and the Jeju Sea, whereas CRI was with fall and the East (Japan) Sea ground. The maturity did not show a distinct pattern. Our results indicate that green turtles in the feeding grounds around the Korean Peninsula originate mainly from the Japan MU and have genetic origins in the Japan, Indo-Pacific, and Central South Pacific clades. Our results provide crucial insights into rookeries and MUs, which are the focus of conservation efforts of the Republic of Korea and potential parties to collaborate for green turtle conservation.

Unoculubranchiobdella sp. (Hirudinea: Ozobranchidae) as a vector for Haemogregarina spp. in freshwater turtles from Brazil.

Species of Haemogregarina are blood parasites known to parasitise vertebrate hosts, including fishes (Haemogregarina sensu lato) and freshwater turtles (Haemogregarina sensu stricto). Their vectors, include gnathiid isopods and leeches, respectively. In turtles, Haemogregarina balli has the best-characterized life cycle in the genus. However, no studies in Brazil have suggested a possible vector for any species of Haemogregarina from freshwater turtles. Therefore, in the present study, we provide insights into a leech vector based on specimens found feeding on two species of freshwater turtles, Podocnemis unifilis and Podocnemis expansa, using morphological and molecular data. In 2017 and 2019, freshwater turtles were collected in Goiás State, Brazil. Hosts were inspected for ectoparasites and leeches were collected from two specimens of P. expansa and nine specimens of P. unifilis. Leeches were subsequently identified as members of the genus Unoculubranchiobdella. Leech histological slides revealed haemogregarine-like structures, similar to post-sporogonic merogony, found near the gills and within the posterior sucker. Molecular analysis of the haemeogregarines resulted in the identification of three species of Haemogregarina: Haemogregarina embaubali, Haemogregarina goianensis, and Haemogregarina brasiliana. Therefore, our findings, based on morphology and DNA data suggest leeches of the genus Unoculubranchiondella as vectors for at least three species of Haemogregarina from Brazilian turtles.

Innovative monitoring scheme adapted to remote, scattered nesting aggregation reveals a major loggerhead turtle rookery in New Caledonia, South Pacific.

The loggerhead turtle Caretta caretta is a large marine turtle with a cosmopolitan repartition in warm and temperate waters of the planet. The South Pacific subpopulation is classified as 'Critically Endangered' on the IUCN Red List, based on the estimated demographic decline. This precarious situation engages an urgent need to monitor nesting populations in order to highlight conservation priorities and to ensure their efficiency over time. New Caledonia encompasses a large number of micro and distant nesting sites, localized on coral islets widely distributed across its large lagoon. Adequately surveying nesting activities on those hard-to-reach beaches can prove to be challenging. As a result, important knowledge gaps prevail in those high-potential nesting habitats. For the first time, an innovative monitoring scheme was conducted to assess the intensity of nesting activities, considered as a proxy of the population size, on an exhaustive set of islets located in the 'Grand Lagon Sud' area. These data were analyzed using a set of statistical methods specially designed to produce phenology and nesting activity estimates using Bayesian methods. This analysis revealed that this rookery hosts a large nesting colony, with a mean annual estimate of 437 nests (95% Credible Interval = 328-582). These numbers exceed that of the previous estimated annual number of loggerhead turtle nests in New Caledonia, highlighting the exceptional nature of this area. Considering the fact that similar high-potential aggregations have been identified in other parts of New Caledonia, but failed to be comprehensively assessed to this day, we recommend carrying out this replicable monitoring scheme to other locations. It could allow a significant re-evaluation of the New Caledonian nesting population importance and, ultimately, of its prevailing responsibility for the protection of this patrimonial yet endangered species.

The first complete mitochondrial genome of the critically endangered Malaysian giant turtle, Orlitia borneensis (Testudines: Geoemydidae).

We present here the complete mitochondrial sequence of the critically endangered Malaysian giant turtle, Orlitia borneensis. The assembled mitochondrial genome includes 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA genes (rRNAs), and one control region. This mitochondrial genome has been archived in the NCBI GenBank with accession number OQ808845. The Batagur control region is relatively smaller than O. borneensis and closer to Aldabrachelys gigantea, which suggests potentially that O. borneensis has undergone an expansion in the control region.

Spatial and life history variation in a trait-based species vulnerability and impact model.

Anthropogenic pressures threaten biodiversity, necessitating conservation actions founded on robust ecological models. However, prevailing models inadequately capture the spatiotemporal variation in environmental pressures faced by species with high mobility or complex life histories, as data are often aggregated across species' life histories or spatial distributions. We highlight the limitations of static models for dynamic species and incorporate life history variation and spatial distributions for species and stressors into a trait-based vulnerability and impact model. We use green sea turtles in the Greater Caribbean Region to demonstrate how vulnerability and anthropogenic impact for a dynamic species change across four life stages. By incorporating life stages into a trait-based vulnerability model, we observed life stage-specific vulnerabilities that were otherwise unnoticed when using an aggregated trait value set. Early life stages were more vulnerable to some stressors, such as inorganic pollution or marine heat waves, and less vulnerable to others, such as bycatch. Incorporating spatial distributions of stressors and life stages revealed impacts differ for each life stage across spatial areas, emphasizing the importance of stage-specific conservation measures. Our approach showcases the importance of incorporating dynamic processes into ecological models and will enable better and more targeted conservation actions for species with complex life histories and high mobility.

Genetic diversity and host specificity of Blastocystis in reptiles, Eastern Thailand.

Blastocystis inhabits the digestive tracts of a diverse range of hosts. Transmission patterns, including host specificity, and the clinical and public health significance of Blastocystis in humans remain poorly understood. This study aimed to investigate the distribution and genetic diversity of Blastocystis in herbivorous and carnivorous reptiles in Eastern Thailand. A total of 501 faecal samples were collected from 363 iguanas, 79 bearded dragons, 50 tortoises, and nine snakes in an animal breeding farm in Chonburi Province, Eastern Thailand. Detection and differentiation of Blastocystis was based on amplification, sequencing, and phylogenetic analysis of specific small subunit (SSU) ribosomal RNA genes from faecal DNA extracted from the samples. Altogether 101/501 samples (20 %) were polymerase chain reaction (PCR) and sequencing-positive for Blastocystis, 90 (89 %) of which were from iguanas; the remaining positive samples were from African spurred tortoise (n=6), Bearded dragon (n=3), Leopard tortoise (n=1), and Red-footed tortoise (n=1). Phylogenetic analysis revealed that most of the Blastocystis sequences from iguanas were largely similar, and they were distinct from those of the tortoises. Subtype 17 was found in the three bearded dragons and likely reflected Blastocystis from prey animals. This is the largest survey of Blastocystis in reptiles to date. Remarkable differences in Blastocystis colonization rates and genetic diversity were observed between iguanas and other reptile orders, and what was considered Blastocystis colonization was only observed in herbivorous reptiles.

Effect of cadmium on histopathological injuries and ultra-structural changes of kidney of the turtle Mauremys reevesii.

This research investigated the effect of cadmium on the tissue and cell of kidney of the turtle Mauremys reevesii. Twenty turtles were injected with cadmium at 0, 7.5, 15, 30 mg/kg separately and five turtles were taken in each group at two weeks after exposure. Kidneys were immediately excised and macroscopic pathological changes were observed, then the kidneys were fixed in 4% paraformaldehyde for histopathological examination and fixed in 2.5% (v/v) glutaraldehyde for examination of ultra-structure. The tissues of kidney presented varying degrees of histopathological lesions in cadmium treated turtles by a dose-dependent manner under the light microscope. Under transmission electron microscope, renal tubules cells presented varying degrees of dose-dependent lesions. The results indicated that cadmium can cause cell damages to the kidney, in particular to the mitochondria. Mitochondria can be used as one biomarker in the monitoring of cadmium pollution and its quantitative risk assessments.

Body size predicts ontogenetic nitrogen stable-isotope (δ15N) variation, but has little relationship with trophic level in ectotherm vertebrate predators.

Large predators have disproportionate effects on their underlying food webs. Thus, appropriately assigning trophic positions has important conservation implications both for the predators themselves and for their prey. Large-bodied predators are often referred to as apex predators, implying that they are many trophic levels above primary producers. However, theoretical considerations predict both higher and lower trophic position with increasing body size. Nitrogen stable isotope values (δ15N) are increasingly replacing stomach contents or behavioral observations to assess trophic position and it is often assumed that ontogenetic dietary shifts result in higher trophic positions. Intraspecific studies based on δ15N values found a positive relationship between size and inferred trophic position. Here, we use datasets of predatory vertebrate ectotherms (crocodilians, turtles, lizards and fishes) to show that, although there are positive intraspecific relationships between size and δ15N values, relationships between stomach-content-based trophic level (TPdiet) and size are undetectable or negative. As there is usually no single value for 15N trophic discrimination factor (TDF) applicable to a predator species or its prey, estimates of trophic position based on δ15N in ectotherm vertebrates with large size ranges, may be inaccurate and biased. We urge a reconsideration of the sole use of δ15N values to assess trophic position and encourage the combined use of isotopes and stomach contents to assess diet and trophic level.

Loggerhead Sea Turtles as Hosts of Diverse Bacterial and Fungal Communities.

Research on microbial communities associated with wild animals provides a valuable reservoir of knowledge that could be used for enhancing their rehabilitation and conservation. The loggerhead sea turtle (Caretta caretta) is a globally distributed species with its Mediterranean population categorized as least concern according to the IUCN Red List of Threatened Species as a result of robust conservation efforts. In our study, we aimed to further understand their biology in relation to their associated microorganisms. We investigated epi- and endozoic bacterial and endozoic fungal communities of cloaca, oral mucosa, carapace biofilm. Samples obtained from 18 juvenile, subadult, and adult turtles as well as 8 respective enclosures, over a 3-year period, were analysed by amplicon sequencing of 16S rRNA gene and ITS2 region of nuclear ribosomal gene. Our results reveal a trend of decreasing diversity of distal gut bacterial communities with the age of turtles. Notably, Tenacibaculum species show higher relative abundance in juveniles than in adults. Differential abundances of taxa identified as Tenacibaculum, Moraxellaceae, Cardiobacteriaceae, and Campylobacter were observed in both cloacal and oral samples in addition to having distinct microbial compositions with Halioglobus taxa present only in oral samples. Fungal communities in loggerheads' cloaca were diverse and varied significantly among individuals, differing from those of tank water. Our findings expand the known microbial diversity repertoire of loggerhead turtles, highlighting interesting taxa specific to individual body sites. This study provides a comprehensive view of the loggerhead sea turtle bacterial microbiota and marks the first report of distal gut fungal communities that contributes to establishing a baseline understanding of loggerhead sea turtle holobiont.

No Ranaviral DNA Found in Australian Freshwater Turtles, 2014-19, Despite Previous Serologic Evidence.

Ranaviruses are pathogens of ectothermic vertebrates (fish, amphibians, and reptiles). Turtles are the most common group of reptiles reported with ranaviral infections. However, there have been no surveys for wild ranaviral infection in any turtles from the suborder Pleurodira, despite ranaviral distributions and experimentally susceptible pleurodiran turtle populations overlapping in several areas, including Australia. We assayed 397 pooled blood samples from six Australian freshwater turtle species collected from five different sites in northern Australia between 2014 and 2019. Historical serologic surveys in the area had found antiranaviral antibodies; however, we did not detect any ranaviral DNA in our samples. Discrepancies between historical serologic and our molecular results may be explained by low viral prevalence during the years that these samples were collected, survivorship bias, or possibly an age class bias in sampling.

Seagrass repression by green turtles (Chelonia mydas) around Taiping Island in the south China sea: Experimental evidence and management insights.

Seagrass meadows serve as critical marine habitats, offering numerous benefits to both humans and wildlife. Taiping Island, situated in the South China Sea, has been historically known for its abundant seagrass meadows. However, satellite imagery suggested there was a low density of seagrasses around Taiping Island. On the other hand, many green turtles (Chelonia mydas) were observed from the shore. To investigate this phenomenon, we conducted drone surveys of the shallow reefs and discovered a density of 902 ± 601 (mean ± SD) individual turtles per square kilometer during high tides. In addition, we conducted a cage experiment to test the hypothesis that large herbivores are impacting seagrass abundance negatively. The results indicated that the blade lengths of seagrasses in cages were significantly longer than those outside. It is likely that large herbivores such as green sea turtles are the key consumers of seagrass on the shallow reef flats of Taiping Island. Accordingly, further research and management should take into account that the increasing number of sea turtles may deplete the seagrasses and have an impact on the seagrass ecosystems.

Roles of estrogen receptors during sexual reversal in Pelodiscus sinensis.

BACKGROUND: The Chinese soft-shelled turtle, Pelodiscus sinensis, exhibits distinct sexual dimorphism, with the males growing faster and larger than the females. During breeding, all-male offspring can be obtained using 17ß-estradiol (E2). However, the molecular mechanisms underlying E2-induced sexual reversal have not yet been elucidated. Previous studies have investigated the molecular sequence and expression characteristics of estrogen receptors (ERs). METHODS AND RESULTS: In this study, primary liver cells and embryos of P. sinensis were treated with ER agonists or inhibitors. Cell incubation experiments revealed that nuclear ERs (nERs) were the main pathway for the transmission of estrogen signals. Our results showed that ERα agonist (ERα-ag) upregulated the expression of Rspo1, whereas ERα inhibitor (ERα-Inh) downregulated its expression. The expression of Dmrt1 was enhanced after ERα-Inh + G-ag treatment, indicating that the regulation of male genes may not act through a single estrogen receptor, but a combination of ERs. In embryos, only the ERα-ag remarkably promoted the expression levels of Rspo1, Wnt4, and ß-catenin, whereas the ERα-Inh had a suppressive effect. Additionally, Dmrt1, Amh, and Sox9 expression levels were downregulated after ERß inhibitor (ERß-Inh) treatment. GPER agonist (G-ag) has a significant promotion effect on Rspo1, Wnt4, and ß-catenin, while the inhibitor G-Inh does not affect male-related genes. CONCLUSIONS: Overall, these results suggest that ERs play different roles during sexual reversal in P. sinensis and ERα may be the main carrier of estrogen-induced sexual reversal in P. sinensis. Further studies need to be performed to analyze the mechanism of ER action.

Hematological and genotoxic biomarkers in a natural population of freshwater turtles Trachemys dorbigni (Duméril & Bibron, 1835) (Testudines: Emydidae) living in an urban area in Southern Brazil.

The present study aimed to report the morphometric and hematological indices and genotoxicity of a free-life population of D'Orbigny's slider turtles (Trachemys dorbigni) living in an urban area in Southern Brazil. For that, 16 specimens were randomly captured in an urban canal that receives irregular releases of wastewater. Biometrics and external visual changes were analyzed, such as turtle shell deformities, and the presence of parasites. Blood samples were collected to evaluate the hematological profile and the presence of micronuclei and other erythrocyte nuclear abnormalities as potential mutagenic and genotoxic effects. Water physicochemical parameters were also measured. Organisms with ectoparasites (31.25%) and small carapace deformations (56.25%) were observed, but maximum carapace length and weight were considered normal for the species according to the literature. The blood profile indicated low hemoglobin and hematocrit and a high number of total leukocytes, particularly eosinophils which characterize parasitic infections. A frequency of 0.12% for the micronucleus was considered basal, but the frequency of other erythrocyte abnormalities was evident, mainly of blebbed nuclei (63.79%), indicating chromosomal damage in the early stage. The results of this study suggest that natural populations of chelonian inhabiting urbanized areas are impacted by anthropogenic activities in the surrounding environment. Furthermore, it provides comprehensive data which can serve as a comparative model for environmental monitoring studies involving turtles.

Comparative analysis for nutrients, flavor compounds, and lipidome revealed the edible value of pond-cultured male Pelodiscus sinensis with different ages.

Pelodiscus sinensis is an aquatic product with a long growth cycle in pond culture and high nutritional value meat. The flavor compounds, nutrients, and lipidome were investigated to explore the edible value changes of turtle meat aged 3 to 6 years (Y3 to Y6). Typically, P. sinensis meat is rich in high-quality protein (EAAI ≥81.22, AAS ≥86.47). Y6 has the highest level of Se, protein, amino acids, and high unsaturated fatty acids, including EPA + DHA. Y5 has the most delicious amino acids, polyunsaturated fatty acids, and key odorant content. The stronger flavor of Y5 may be mainly related to C18:2n6t and C18:2n6c. Further, triacylglycerols (TAG) and phosphatidylcholine (PC) were significant changes in Y5. Additionally, PI (16:0/18:1) was identified as the potential biomarker. These results provided available information on P. sinensis marketing age and revealed the potential impact of nutrients on the formation of VOCs.

Accumulation rate and sources of plastic marine litter at nesting grounds of green turtles on the North Island of Qilianyu, Xisha Islands, South China Sea.

In this study, the accumulation rate of plastic litter was investigated by sampling quadrats placed on the North Island of Qilianyu, and the composition was analyzed and identified to determine its source. The results showed that the annual average accumulation rate of plastic litter on North Island was 0.64 ± 0.32 pieces·m-2·month-1, with a mass accumulation rate of 11.30 ± 7.73 g·m-2·month-1. The accumulation rate of plastic litter was mainly influenced by wind speed and direction, with higher accumulation rates occurring during the southwest monsoon season and tropical cyclones. ATR-FTIR analysis indicated that polyethylene (44 %) and polypropylene (41 %) were the most abundant types of polymers. This study reveals the current status of plastic litter pollution in green turtle nesting grounds on North Island in Qilianyu, which can be used as a reference for management strategies that mitigate plastic litter pollution.

Exposure to high concentrations of triphenyl phosphate altered functional performance, liver metabolism and intestinal bacterial composition of aquatic turtles.

Organophosphorus flame retardants, such as triphenyl phosphate (TPhP), exist ubiquitously in various environments owing to their widespread usage. Potential toxic effects of residual flame retardants on cultured non-fish species are not concerned commonly. TPhP-induced physiological and biochemical effects in an aquatic turtle were evaluated here by systematically investigating the changes in growth and locomotor performance, hepatic antioxidant ability and metabolite, and intestinal microbiota composition of turtle hatchlings after exposure to different TPhP concentrations. Reduced locomotor ability and antioxidant activity were only observed in the highest concentration group. Several metabolic perturbations that involved in amino acid, energy and nucleotide metabolism, in exposed turtles were revealed by metabolite profiles. No significant among-group difference in intestinal bacterial diversity was observed, but the composition was changed markedly in exposed turtles. Increased relative abundances of some bacterial genera (e.g., Staphylococcus, Vogesella and Lawsonella) probably indicated adverse outcomes of TPhP exposure. Despite having only limited impacts of exposure at environmentally relevant levels, our results revealed potential ecotoxicological risks of residual TPhP for aquatic turtles considering TPhP-induced metabolic perturbations and intestinal bacterial changes.

Conservation units alone are insufficient to protect Brazilian Amazonian chelonians.

The creation of protected areas (PAs) is not always based on science; consequently, some aquatic species may not receive the same level of protection as terrestrial ones. The objective of this study was to identify priority areas for the conservation of chelonians in the Brazilian Amazon basin and assess the contribution of PAs, distinguishing between Full Protection Areas, Sustainable Use Areas, and Indigenous Lands for group protection. The entire species modeling procedure was carried out using Species Distribution Models. Location records were obtained from platforms such as SpeciesLink, GBIF, the Hydroatlas database, and WorldClim for bioclimatic variables adjusted with algorithms like Maximum Entropy, Random Forest, Support Vector Machine, and Gaussian-Bayesian. Indigenous lands cover more than 50% of the distribution areas of chelonian species in the Brazilian Amazon. Protected areas with higher conservation importance (Full Protection Areas and Sustainable Use Areas) hold less than 15% of the combined species distribution. Researchers face significant challenges when making decisions with models, especially in conservation efforts involving diverse taxa that differ significantly from one another within a group of individuals.

First evidence of marine turtle gastroliths in a fossil specimen: Paleobiological implications in comparison to modern analogues.

Semi-articulated remains of a large chelonioid turtle from the Turonian strata (Upper Cretaceous; ca. 93.9-89.8 Myr) near Sant'Anna d'Alfaedo (Verona province, northeastern Italy) are described for the first time. Together with the skeletal elements, the specimen also preserves pebbles inside the thoracic area which are lithologically distinct from the surrounding matrix. These allochthonous clasts are here interpreted as geo-gastroliths, in-life ingested stones that resided in the digestive tract of the animal. This interpretation marks the first reported evidence of geophagy in a fossil marine turtle. SEM-EDS analysis, together with macroscopic petrological characterization, confirm the presence of both siliceous and carbonatic pebbles. These putative geo-gastroliths have morphometries and size ranges more similar to those of gastroliths in different taxa (fossils and extant) than allochthonous "dropstone" clasts from the same deposit that were carried by floating vegetation A dense pitted pattern of superficial erosion is microscopically recognizable on the carbonatic gastroliths, consistent with surface etching due to gastric acids. The occurrence of a similar pattern was demonstrated by the experimental etching of carbonatic pebbles with synthetic gastric juice. Gut contents of modern green sea turtles (Chelonia mydas) were surveyed for substrate ingestion, providing direct evidence of geophagic behavior in extant chelonioids. Comparison with modern turtle dietary habits may suggests that the pebbles were ingested as a way to supplement calcium after or in preparation for egg deposition, implying that the studied specimen was possibly a gravid female.

Embryonic development of the scorpion mud turtle (Kinosternon scorpioides) bred in captivity.

Background: The developmental biology of Kinosternon scorpioides is described, based on the phenotype. This species is important for the flora because they are excellent seed disseminators. In addition, basic embryological information is not yet fully clarified, and this research provides unprecedented information on the chelonian embryology of the Amazonian fauna. Aim: The present study aims to identify the embryology of K. scorpioides in captivity during different periods. Methods: Females were monitored throughout the reproductive cycle, by video monitoring, to identify nests and the presence of newly laid eggs. At regular weekly intervals, embryo samples were collected fixed in a 4% paraformol solution and preserved in 70% alcohol. For the embryonic characterization, we used a stereomicroscope and the scanning electron microscopy method. Results: We describe 15 embryonic stages for a 15-week (105-day) incubation process. Only at 42 days (6th week) was the morphological characterization of a chelonian observed and at the 12th week (Stage XII), the phenotypic characterization of the species K. scorpioides. Conclusion: In view of the evidence, we found that these phases are similar to the other turtles, with structural variations in the appearance and disappearance of structures due to the specific characteristics of the species.