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Classifying animal behaviors in their natural environments is both challenging and ecologically important, but the use of biologgers with multiple sensors has significantly advanced this research beyond the capabilities of traditional methods alone. Here, we show how biologgers containing an integrated tri-axial accelerometer, GPS logger and immersion sensor were used to infer behavioural states of a cryptic, freshwater turtle, the Blanding's turtle (Emydoidea blandingii). Biologgers were attached to three males and five females that reside in two undisturbed coastal marshes in northeastern Georgian Bay (Ontario, Canada) between May and July 2023. Raw acceleration values were separated into static and dynamic acceleration and subsequently used to calculate overall dynamic body acceleration (ODBA) and pitch. The unsupervised Hidden Markov Model (HMM) successfully differentiated five behavioural states as follows: active in water, resting in water, active out of water, resting in water, and nesting. Overall accuracy of the classification was 93.8%, and except for nesting (79%), all other behaviours were above 92%. There were significant differences in daily activity budgets between male and female turtles, with females spending a greater proportion of time active out of water, and inactive out of the water, while males spent a greater proportion of time active in water. These differences were likely a result of large seasonal life-history requirements such as nesting and mate finding. Accurate classification of behavioural states is important for researchers to understand fine-scale activities carried out during the active season and how environmental variables may influence the behaviours of turtles in their natural habitats. [ABSTRACT FROM AUTHOR]

Reliable population estimates are important for making informed management decisions about wildlife species. Standardized survey protocols have been developed for monitoring population trends of the wood turtle (Glyptemys insculpta), a semi-aquatic freshwater turtle species of conservation concern throughout its distribution in east-central North America. The protocols use repeated active search surveys of defined areas, allowing for estimation of survey-specific detection probability (p) and site-specific abundance. These protocols assume population closure within the survey area during the survey period, which is unlikely to be met as wood turtles are a highly mobile species. Additionally, current protocols use a single-pass design that does not allow for separation of availability (pa) and detectability (pd). If there are systematic influences on pa or pd that are not accounted for in the survey design or data analysis, then resulting abundance estimates could be biased. The objectives of this study were to determine if pa is a random process and if pa and pd are influenced by demographic characteristics. We modified the wood turtle survey protocol used in the upper Midwest to include a double-pass design, allowing us to estimate pa and pd using a robust design capture-recapture model. The modified protocol was implemented at 14 wood turtle monitoring sites in Minnesota and Wisconsin between 2017 and 2022. Our results indicated that pa was non-random and that pd increased with turtle carapace length. Our study suggests that model assumptions for current wood turtle population models may be violated, likely resulting in an overestimation of abundance. We discuss possible protocol and modeling modifications that could result in more accurate wood turtle abundance estimates. [ABSTRACT FROM AUTHOR]

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. [ABSTRACT FROM AUTHOR]

Verifying habitats, including the foraging and nesting areas for sea turtles, enables an understanding of their spatial ecology and successful planning of their conservation and management strategies. Recently, the observation frequency and bycatch of loggerhead (Caretta caretta) and green (Chelonia mydas) turtles have increased in the northern limit of their distribution range, in the northern part of the East China Sea and East (Japan) Sea. We conducted satellite tracking to investigate the habitat use of seven loggerhead and eight green turtles from June 2016 to August 2022 in this area, where little is known about their spatial ecology. We applied a 50 percent volume contour method to determine their main foraging areas and analyzed 6 environmental variables to characterize their habitats. Loggerhead turtles mainly stayed in and used the East China Sea as a foraging area during the tracking period, while two individuals among them also used the East Sea as a seasonal foraging area. Most green turtles also used the East China Sea as a foraging area, near South Korea and Japan, with one individual among them using the lower area of the East Sea as a seasonal foraging area. Notably, one green turtle traveled to Hainan Island in the South China Sea, a historical nesting area. Our results showed that the two sea turtle species included the East Sea as a seasonal foraging area, possibly owing to the abundance of food sources available, despite its relatively lower sea temperature. Considering that loggerhead and green sea turtles were observed using the northern part of the East China Sea and East Sea more frequently than previously known and that the sea temperature gradually increases due to climate change, conservation and management activities are required for sea turtles in these areas. [ABSTRACT FROM AUTHOR]

Impressions of vertebrate bodies or their parts, such as trace fossils and natural molds of bones, are a valuable source of information about ancient faunas which may supplement the standard fossil record based on skeletal elements. Whereas trace fossils of animal activity are relatively common and actively studied within the field of ichnology, and natural impressions of internal or external surfaces are a frequent preservation mode in fossil invertebrates, natural molds of bones are comparatively rare and less extensively documented and discussed. Among them, internal molds (steinkerns) of turtle shells are a relatively well-known form of preservation, but the mechanisms and taphonomic prerequisites leading to their formation are poorly studied. External shell molds are even less represented in the literature. Herein, we describe a historic specimen of a natural external turtle plastron mold from the Triassic (Norian) Löwenstein Formation of Germany–a formation which also yielded a number of turtle steinkerns. The specimen is significant not only because it represents an unusual form of preservation, but also due to its remarkably large size and the presence of a potential shell pathology. Although it was initially interpreted as Proterochersis sp., the recent progress in the knowledge of proterochersid turtles leading to an increase in the number of known taxa within that group allows us to verify that assessment. We confirm that the specimen is morphologically consistent with the genus and tentatively identify it as Proterochersis robusta, the only representative of that genus from the Löwenstein Formation. We note, however, that its size exceeds the size observed thus far in Proterochersis robusta and fits within the range of Proterochersis porebensis from the Grabowa Formation of Poland. The marks interpreted as shell pathology are morphologically consistent with Karethraichnus lakkos–an ichnotaxon interpreted as a trace of ectoparasites, such as leeches. This may support the previously proposed interpretation of Proterochersis spp. as a semiaquatic turtle. Moreover, if the identification is correct, the specimen may represent a very rare case of a negative preservation of a named ichnotaxon. Finally, we discuss the taphonomy of the Löwenstein Formation turtles in comparison with other Triassic turtle-yielding formations which show no potential for the preservation of internal or external shell molds and propose a taphonomic model for the formation of such fossils. [ABSTRACT FROM AUTHOR]

Freshwater fauna is facing an uphill task for survival in the Ganga Basin, India, due to a range of factors causing habitat degradation and fragmentation, necessitating conservation interventions. As part of the ongoing efforts to conserve the freshwater fauna of the Basin, we are working on rehabilitating rescued freshwater chelonians. We carry out various interventions to restore rescued individuals to an apparent state of fitness for their release in suitable natural habitats. Morphometric measurements are crucial to managing captive wild animals for assessing their growth and well-being. Measurements are made using manual methods like vernier caliper that are prone to observer error experience and require handling the specimens for extended periods. Digital imaging technology is rapidly progressing at a fast pace and with the advancement of technology. We acquired images of turtles using smartphones along with manual morphometric measurements using vernier calipers of the straight carapace length and straight carapace width. The images were subsequently processed using ImageJ, a freeware and compared with manual morphometric measurements. A significant decrease in the time spent in carrying out morphometric measurements was observed in our study. The difference in error in measurements was, however, not significant. A probable cause for this may have been the extensive experience of the personnel carrying out the measurements using vernier caliper. Digital image processing technology can cause a significant reduction in the stress of the animals exposed to handling during measurements, thereby improving their welfare. Additionally, this can be used in the field to carry out morphometric measurements of free-ranging individuals, where it is often difficult to capture individuals, and challenges are faced in obtaining permission to capture specimens. [ABSTRACT FROM AUTHOR]

Few areas within the Great Lakes basin are currently free from impact of human activities, and it is important to study these reference conditions for comparison with degraded sites in those regions. Here, we use radio telemetry to investigate habitat use, movement, and habitat selection of a population of the endangered (Federally in Canada) Blanding's turtle (Emydoidea blandingii, BLTU) inhabiting a mostly undisturbed archipelago located at the northern shore of Mnidoo gamii (Georgian Bay), Ontario over two active seasons (May to September 2021 and 2022). We found a mean home range of 16.21 ha for females (n = 7) and 15.10 ha for males (n = 7). Of the five habitat classes (Marsh, Open Water, Rock, Peatland, and Forest), females used all except Peatland during the nesting season, and both sexes used all five habitat classes throughout both active seasons in 2021 and 2022. Disproportionate habitat use was detected at the landscape scale but not at the home range scale which was consistent with the hypothesis that adult Blanding's turtles residing in relatively undisturbed sites with abundant habitat types use all habitat types according to their availability. We also observed the use of open, deep water by Blanding's Turtles as travel corridors for nesting and mating. Effective future conservation strategies should prioritize the protection and connectivity of relatively undisturbed wetlands, forests, and rock barrens in this region and use this study as a reference condition to compare BLTU habitat use and movement across disturbance gradients within Georgian Bay. [ABSTRACT FROM AUTHOR]

Long-term monitoring of sea turtle aggregations is critical for understanding the impacts of environmental changes on their population health and habitat suitability. Brunei Bay is a significant foraging ground for green turtles in the South China Sea. We analyzed the body size, hematology and body condition of green turtles for their health status in their foraging ground in Brunei Bay over a period of nine years (2011–2019). Additionally, we used mitochondrial DNA (mtDNA) to evaluate changes in the size and genetic composition of green turtle aggregations. Our findings revealed that the size composition of the green turtle population varied seasonally, but there were no significant temporal changes in genetic and size compositions. Hematology parameters and Fulton's body condition index were consistent with those reported for apparently healthy green turtles. Furthermore, we found that blood reference intervals indicated the turtles were healthy. These results provide valuable baseline data for future comparisons with other foraging aggregations and for long-term monitoring of green turtles in Brunei Bay. [ABSTRACT FROM AUTHOR]

The limestones of the Solnhofen area in southern Germany are one of the most important fossil Lagerstätten from the entire Mesozoic era, especially famous for the exquisitely preserved vertebrates. The turtles from the Solnhofen Limestone have been always of special interest because they include some of the best-preserved specimens from the Mesozoic. Here, we describe a new turtle specimen from the Torleite Formation (Kimmeridgian) of Painten and refer it to the thalassochelydian turtle Solnhofia parsonsi based on the presence of a unique combination of characters. The far majority of morphological differences from previously published specimens can be explained by ontogeny as the new specimen represents a larger, more ossified, and presumably older individual. Additionally, the specimen from Painten is the first described specimen of S. parsonsi preserving the largely complete and articulated limbs, the preservation of which indicates that the taxon did not possess stiffened paddles present in more pelagic marine turtles and is consistent with a previously inferred nearshore marine lifestyle. Contrary to previous inferences, we argue that taphonomic preservation of digits in articulated fossil turtles from laminated deposits cannot be used alone to infer marine or freshwater habitat. Finally, the new specimen from Painten is only the second, for which detailed information on its stratigraphic position and locality of origin are known. [ABSTRACT FROM AUTHOR]

Online sales are increasingly a route by which exotic animals are sold in the global pet trade. There are numerous types of online platforms and transaction types, and dedicated classified advertisement sites are a popular means of buying and selling animals. Despite their large and increasing use, we have a relatively poor understanding of the number of, and taxonomic variation in, the animals sold online. This information may be key in efforts to optimise the welfare of the animals being sold, and the ethics and sustainability of the trade via that platform. To fill this knowledge gap, we monitored and analysed the advertisements of chelonians (turtles and tortoises) placed on one of the United Kingdom's largest dedicated classified ads sites, www.pets4homes.co.uk, over the course of a year, from July 2020 until June 2021. We analysed temporal, taxonomic, and advertiser related trends in the volumes of advertisements placed and compared the prices and the sentiment of language within adverts for different species. We found that the species advertised, the prices requested, and infrequent use of the site by most advertisers is consistent with most adverts being for animals being resold by casual users. Further, we found that turtles were consistently advertised for lower prices and in multiples than tortoises, and that the language with which they were advertised was less positive. We conclude that on this website the online trade reflects the broader trade, rather than drives the sales of chelonians in the UK, and that any interventions aiming to improve welfare and sustainability would be better placed earlier in the supply chain. [ABSTRACT FROM AUTHOR]

Freshwater turtle populations are declining globally as a result of anthropogenic activities. Threats to turtles in urban areas are exacerbated by road mortality and subsidized predators, which can lead to catastrophic shifts in population size and structure. Headstarting is used as a conservation tool to supplement turtle populations that may otherwise face extirpation. A headstarting program began in 2012 to recover a functionally extinct population of Blanding's Turtles (Emydoidea blandingii) 26in Rouge National Urban Park (RNUP), Ontario, Canada. The original population included five adults and one juvenile turtle. From 2014 to 2020, 270 headstarted turtles were released. The population has been monitored annually since 2014 using visual-encounter surveys, radio-telemetry, and live trapping (from 2018 onwards). We used mark-recapture and radio-telemetry data to quantify abundance, survival, and sex ratio of the headstarted turtle population. Using a Jolly-Seber model, we estimated abundance to be 183 turtles (20 turtles/ha) in 2020. Estimated survival of headstarted turtles approached 89%, except for turtles released in 2019 when survival was 43% as a result of a known mass mortality event at the study site. Pre- and post-release sex ratios were not significantly different (χ2 = 1.92; p = 0.16), but shifted from 1:1.5 to 1:1 male:female post-release. Given that the headstarted turtles have not yet reached sexual maturity, it is unclear whether headstarted turtles will reach adulthood and successfully reproduce to maintain a self-sustaining population. Thus, to evaluate the success of the headstarting program, long-term monitoring is required. [ABSTRACT FROM AUTHOR]

Alien invasive species are posing conservation challenges worldwide. Pet trade, one of the many ways, is worsening the situation. Especially, pet turtles have been released into nature due to their longer life span and peoples' religious and traditional beliefs. In addition, unwanted and undesired pets are also released. While information on the successful local establishment and subsequent dispersal into new habitats is required to designate an invasive and ecosystem-disturbing species, alien freshwater turtle nests have always been hard to find and identify in nature. Because one should identify nests by the eggs, which do not always guide properly, as adults abandon the sites quickly. We thought the recent advancement in DNA technology may help improve the situation. We studied Pseudemys peninsularis, one of the most traded freshwater turtle pet species, which has already been reported from a wide range of wild areas in South Korea. Yet, it is not designated as ecosystem-disturbing species due to a lack of adequate information on their local reproduction and establishment. We conducted surveys and found two nests in Jeonpyeongje Neighborhood Park, Maewol-dong, Seo-gu, Gwangju. We developed the methodology for extracting DNA from the eggshells and successfully identified the nests by phylogenetic analysis and verified through egg characteristics and morphological features of artificially hatched juveniles. This was the first successful initiative to extract DNA from freshwater turtle eggshells. We believe it will help future researchers identify the alien invasive turtle nests and develop their control and management policies. In addition, our study also included comparative descriptions and schematic diagrams of the eggs of eight freshwater turtles, including a native and three ecosystem-disturbing species, from South Korea. We urged an immediate designation of P. peninsularis as an ecosystem-disturbing species considering its local establishment, distribution range, and potential negative impact on native ecosystems. [ABSTRACT FROM AUTHOR]

In the United States (U.S.), pet turtles have been associated with outbreaks of salmonellosis, a serious and sometimes-fatal intestinal illness caused by Salmonella bacteria, with nearly 300,000 people being infected in some years. Children are particularly susceptible because of their propensity to put items, including small turtles, in their mouths. In 1975, a U.S. federal regulation prohibited the sale of turtles <4 inches (101.6 mm) in size, except for the purposes of export, scientific, or educational purposes. This regulation was established to reduce the incidence of salmonellosis, particularly in small children. Previous research has not evaluated the availability of turtles <4 inches in size on websites selling wildlife. We monitored 16 websites in 2021 and quantified listings of small turtles. We determined whether information on Salmonella, the 1975 federal regulation, or related state regulations were provided on the websites and determined legality of sales of small turtles by state regulations. We found that all 16 websites openly advertised and sold turtles <4 inches in size, but only half of these websites provided information about Salmonella and/or the federal regulation. These websites required buyers to confirm that they were not purchasing a turtle as a pet, thereby putting the onus on the consumer to adhere to the regulation. We documented 515 listings of turtles <4 inches in size, including 47 species and one hybrid. Our study has demonstrated that internet sales of small turtles currently represent part of the thriving online pet trade in the U.S. Enforcement of the federal regulation faces jurisdictional challenges in most states. Therefore, we recommend continued public education campaigns by public health agencies in the U.S. to help reduce the risk that pet turtle ownership presents. [ABSTRACT FROM AUTHOR]

Research in ecology often requires robust assessment of animal behaviour, but classifying behavioural patterns in free-ranging animals and in natural environments can be especially challenging. New miniaturised bio-logging devices such as accelerometers are increasingly available to record animal behaviour remotely, and thereby address the gap in knowledge related to behaviour of free-ranging animals. However, validation of these data is rarely conducted and classification model transferability across closely-related species is often not tested. Here, we validated accelerometer and water sensor data to classify activity states in two free-ranging freshwater turtle species (Blanding's turtle, Emydoidea blandingii, and Painted turtle, Chrysemys picta). First, using only accelerometer data, we developed a decision tree to separate motion from motionless states, and second, we included water sensor data to classify the animal as being motionless or in-motion on land or in water. We found that accelerometers separated in-motion from motionless behaviour with > 83% accuracy, whereas models also including water sensor data predicted states in terrestrial and aquatic locations with > 77% accuracy. Despite differences in values separating activity states between the two species, we found high model transferability allowing cross-species application of classification models. Note that reducing sampling frequency did not affect predictive accuracy of our models up to a sampling frequency of 0.0625 Hz. We conclude that the use of accelerometers in animal research is promising, but requires prior data validation and development of robust classification models, and whenever possible cross-species assessment should be conducted to establish model generalisability. [ABSTRACT FROM AUTHOR]

The concentrations of trace elements including As, Zn, Cu, Se, Pb, Hg and Cd, were determined in the blood of nesting Kemp's ridley turtles (Lepidochelys kempii) at Rancho Nuevo sanctuary, Tamaulipas, Mexico during 2018–2020. The sequential concentrations analyzed were Zn> Se> Cu> As> Pb; while Cd and Hg concentrations were below the limits of detection (0.01 μg g-1). No significant differences were observed between the concentrations of trace elements (p> 0.05) by year, except Se levels, possibly resulting from recorded seasonal differences in turtle size. No relationships among turtle size vs elements concentration were observed. In conclusion, essential and toxic trace elements concentrations in the blood of nesting Kemp's ridley turtles may be a reflex of the ecosystem in which the turtles develop, that is, with low bioavailability of elements observed in the trophic webs in the Gulf of Mexico. [ABSTRACT FROM AUTHOR]

Current understanding of sea turtle nesting, hatching, and emergence events has been largely limited to observable events on the surface of the sand, though recent approaches using audio or visual equipment have allowed scientists to better understand some underground nest phenomena. We used a technology-based approach to define motion-related Caretta caretta hatching and emergence nest events. We describe a novel low-cost, accelerometer-based system called TurtleSense that can detect movement and temperature within sea turtle nests remotely. TurtleSense is successfully able to specifically detect motion within sea turtle nests over the entire course of incubation. This system allows for the identification of infertile nests and the detection of four predictable sequential developmental activity patterns in viable nests, including a hatch and posthatch period, the timing of which can be used to tightly predict hatchling emergence events almost to the day. TurtleSense provides a much better understanding about what is happening in the nest before emergence and allows for the generation of a theory of the mechanism that triggers mass emergence. Our results suggest that motion plays a large role in hatchling communication and that the timing of emergence events may be related to the cessation of movement within the nest. Current management of sea turtle nesting events is primarily driven by counting the number of days since the nest was laid, with further safeguards placed at the nest upon subsequent visual observation of depression or emergence events. Use of TurtleSense technology can impact nest management and conservation efforts, allowing organizations to use this motion data to more tightly predict emergence dates for sea turtle hatchlings and to use viability data to inform nest management decisions. [ABSTRACT FROM AUTHOR]

Sea turtles exhibit high migratory rates and occupy a broad range of habitats, which in turn makes monitoring these taxa challenging. Applying deep learning (DL) models to vast image datasets collected from citizen science programs can offer promising solutions to overcome the challenge of monitoring the wide habitats of wildlife, particularly sea turtles. Among DL models, object detection models, such as the You Only Look Once (YOLO) series, have been extensively employed for wildlife classification. Despite their successful application in this domain, detecting objects in images with complex backgrounds, including underwater environments, remains a significant challenge. Recently, instance segmentation models have been developed to address this issue by providing more accurate classification of complex images compared to traditional object detection models. This study compared the performance of two state-of-the-art DL methods namely; the object detection model (YOLOv5) and instance segmentation model (YOLOv5-seg), to detect and classify sea turtles. The images were collected from iNaturalist and Google and then divided into 64% for training, 16% for validation, and 20% for test sets. Model performance during and after finishing training was evaluated by loss functions and various indexes, respectively. Based on loss functions, YOLOv5-seg demonstrated a lower error rate in detecting rather than classifying sea turtles than the YOLOv5. According to mean Average Precision (mAP) values, which reflect precision and recall, the YOLOv5-seg model showed superior performance than YOLOv5. The mAP0.5 and mAP0.5:0.95 for the YOLOv5 model were 0.885 and 0.795, respectively, whereas for the YOLOv5-seg, these values were 0.918 and 0.831, respectively. In particular, based on the loss functions and classification results, the YOLOv5-seg showed improved performance for detecting rather than classifying sea turtles compared to the YOLOv5. The results of this study may help improve sea turtle monitoring in the future., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Baek et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Few aquatic animal negative-sense RNA viruses have been characterized, and their role in disease is poorly understood. Here, we describe a virus isolated from diseased freshwater turtles from a Florida farm in 2007 and from an ongoing epizootic among free-ranging populations of Florida softshell turtles (Apalone ferox), Florida red-bellied cooters (Pseudemys nelsoni), and peninsula cooters (Pseudemys peninsularis). Affected turtles presented with similar neurological signs, oral and genital ulceration, and secondary microbial infections. Microscopic lesions were most severe in the softshell turtles and included heterophilic/histiocytic meningoencephalitis, multi-organ vasculitis, and cytologic observation of leukocytic intracytoplasmic inclusions. The virus was isolated using Terrapene heart (TH-1) cells. Ultrastructurally, viral particles were round to pleomorphic and acquired an envelope with prominent surface projections by budding from the cell membrane. Viral genomes were sequenced from cDNA libraries of two nearly identical isolates and determined to be bi-segmented, with an ambisense coding arrangement. The larger segment encodes a predicted RNA-directed RNA polymerase (RdRP) and a putative zinc-binding matrix protein. The smaller segment encodes a putative nucleoprotein and an envelope glycoprotein precursor (GPC). Thus, the genome organization of this turtle virus resembles that of arenaviruses. Phylogenetic analysis shows that the RdRP of the turtle virus is highly diverged from the RdRPs of all known negative-sense RNA viruses and forms a deep branch within the phylum Negarnaviricota, that is not affiliated with any known group of viruses, even at the class level. In contrast, the GPC protein of the turtle virus is confidently affiliated with homologs from a distinct group of fish hantaviruses. Thus, the turtle virus is expected to become the founder of a new taxon of negative-sense RNA viruses, at least with a family rank, but likely, an order or even a class. These viruses probably evolved either by reassortment or by intrasegment recombination between a virus from a distinct branch of negarnaviruses distant from all known groups and a hanta-like aquatic virus. We suggest the provisional name Tosoviridae for the putative new family, with Turtle fraservirus 1 (TFV1) as the type species within the genus Fraservirus. A conventional RT-PCR assay, targeting the TFV1 RdRP, confirmed the presence of viral RNA in multiple tissues and exudates from diseased turtles. The systemic nature of the TFV1 infection was further supported by labeling of cells within lesions using in situ hybridization targeting the RNA of the TFV1 RdRP. Author summary: Freshwater turtles are an important part of aquatic ecosystems, but relatively little is known about diseases that affect them. We discovered a new virus associated with die-offs involving multiple turtle species in Florida. Affected turtles had inflammation involving the nervous system and other organs indicative of viral infection. We isolated a virus from diseased turtles and characterized its morphology using electron microscopy. We also sequenced its entire genome to explore its relatedness to other viruses and developed a PCR-based detection assay. The morphology of the virus particle and genome characteristics were reminiscent of an arenavirus. However, phylogenetic analyses of two viral proteins indicate that the turtle virus most likely evolved through genetic mixing between a distinct branch of RNA viruses distant from all known groups and an aquatic hantavirus. Thus, the turtle virus is expected to become a founding member of a new RNA virus family. We provide evidence linking this novel virus with disease in both wild and captive turtles. [ABSTRACT FROM AUTHOR]

We evaluate the efficacy of environmental DNA (eDNA) techniques to locate wild populations and estimate the population size of the endangered big-headed turtle (Platysternon megacephalum) in Hong Kong. The results from this study are important for identifying priority sites for protection and further research. Additionally, we assess the impact of two environmental variables (temperature and pH) on eDNA quantity. We surveyed 34 streams for three years, sampling four times each year. Four new populations were first identified with eDNA analysis, and then verified by field surveys. Our multi-year survey highlights that eDNA detection can be inconsistent over time, even in streams with known populations. There was no significant relationship between eDNA quantity and the environmental variables tested. Lastly, our results suggest that eDNA methods remain promising to estimate population size, since number of positive detections were positively correlated with population size in streams with known populations. We conclude that eDNA methods are powerful, but care must be taken when interpreting field results as they are affected by species ecology and environmental conditions. [ABSTRACT FROM AUTHOR]

Turtle body size is associated with demographic and other traits like mating success, reproductive output, maturity, and survival. As such, growth analyses are valuable for testing life history theory, demographic modeling, and conservation planning. Two important but unsettled research areas relate to growth after maturity and growth rate variation. If individuals exhibit indeterminate growth after maturity, older adults may have an advantage in fecundity, survival, or both over younger/smaller adults. Similarly, depending on how growth varies, a portion of the population may mature earlier, grow larger, or both. We used 23-years of capture-mark-recapture data to study growth and maturity in the Spotted Turtle (Clemmys guttata), a species suffering severe population declines and for which demographic data are needed for development of effective conservation and management strategies. There was strong support for models incorporating sex as a factor, with the interval growth model reparametrized for capture-mark-recapture data producing later mean maturation estimates than the age-based growth model. We found most individuals (94%) continued growing after maturity, but the instantaneous relative annual plastral growth rate was low. We recommend future studies examine the possible contribution of such slow, continued adult growth to fecundity and survival. Even seemingly negligible amounts of annual adult growth can have demographic consequences affecting the population vital rates for long-lived species. [ABSTRACT FROM AUTHOR]

Human activities have been changing the global biogeographic patterns by the introductions of invasive species. For reptiles, the invasion rate increase of non-native species is remarkably related to the pet trade, especially for freshwater turtles. Here we estimated the invasive potential of the South American turtle Trachemys dorbigni in the Americas using a combination of climatic and human activity variables. We built species distribution models based on data from the native and invasive ranges, using the ensemble model from five different algorithms (GAM, MAXENT, BRT, RF and GBM). We compared the two models' performance and predictions, one calibrated with only climatic variables (climate-driven), and the second also included a descriptive variable of human activity (climate plus human-driven). Suitable areas for T. dorbigni covered occurrence areas of its congeners and highly diversified ecoregions, such as the eastern USA, the islands of Central America, and the south eastern and eastern Brazilian coast. Our results indicate that human activities allow T. dorbigni to establish populations outside of its original climatic niche. Including human activity variables proved fundamental to refining the results to identify more susceptible areas to invasion and to allow the efficient targeting of prevention measures. Finally, we suggested a set of actions to prevent T. dorbigni becoming a highly impacting species in the areas identified as more prone to its invasion. [ABSTRACT FROM AUTHOR]

Conservation concerns are increasing for numerous freshwater turtle species, including Pseudemys gorzugi, which has led to a call for more research. However, traditional sampling methodologies are often time consuming, labor intensive, and invasive, restricting the amount of data that can be collected. Biases of traditional sampling methods can further impair the quality of the data collected, and these shortfalls may discourage their use. The use of unmanned aerial vehicles (UAVs, drones) for conducting wildlife surveys has recently demonstrated the potential to bridge gaps in data collection by offering a less labor intensive, minimally invasive, and more efficient process. Photographs and video can be obtained by camera attachments during a drone flight and analyzed to determine population counts, abundance, and other types of data. In this study we developed a detailed protocol to survey for large, freshwater turtle species in an arid, riverine landscape. This protocol was implemented with a DJI Matrice 600 Pro drone and a SONY ILCE α6000 digital camera to determine P. gorzugi and sympatric turtle species occurrence across 42 sites in southwestern Texas, USA. The use of a large drone and high-resolution camera resulted in high identification percentages, demonstrating the potential of drones to survey for large, freshwater turtle species. Numerous advantages to drone-based surveys were identified as well as some challenges, which were addressed with additional refinement of the protocol. Our data highlight the utility of drones for conducting freshwater turtle surveys and provide a guideline to those considering implementing drone-mounted high-resolution cameras as a survey tool. [ABSTRACT FROM AUTHOR]

Creating a routing backbone is a fundamental problem in both biology and engineering. The routing backbone of the trail networks of arboreal turtle ants (Cephalotes goniodontus) connects many nests and food sources using trail pheromone deposited by ants as they walk. Unlike species that forage on the ground, the trail networks of arboreal ants are constrained by the vegetation. We examined what objectives the trail networks meet by comparing the observed ant trail networks with networks of random, hypothetical trail networks in the same surrounding vegetation and with trails optimized for four objectives: minimizing path length, minimizing average edge length, minimizing number of nodes, and minimizing opportunities to get lost. The ants' trails minimized path length by minimizing the number of nodes traversed rather than choosing short edges. In addition, the ants' trails reduced the opportunity for ants to get lost at each node, favoring nodes with 3D configurations most likely to be reinforced by pheromone. Thus, rather than finding the shortest edges, turtle ant trail networks take advantage of natural variation in the environment to favor coherence, keeping the ants together on the trails. Author summary: We investigated the trail networks of arboreal turtle ants in the canopy of the tropical forest, to ask what characterizes the colony's choice of foraging paths within the vegetation. We monitored day to day changes in the junctions and edges of trail networks of colonies in the dry forest of western Mexico. We compared the paths used by the ants to simulated random paths in the surrounding vegetation. We found that the paths of turtle ants prioritize coherence, keeping ants together on the trail, over minimizing the average edge length. The choice of paths reduces the number of junctions in the trail where ants could get lost, and favors junctions with a physical configuration that makes it likely that successive ants will reinforce the same path. Our work suggests that design principles that emphasize keeping information flow constrained to streamlined, coherent trails may be useful in human-designed distributed routing and transport networks or robot swarms. [ABSTRACT FROM AUTHOR]

For ectotherms, behaviour and associated energetic costs are directly related to thermal conditions. In the present context of global change, estimating time-budget for these species is relevant to assess and predict their capacity to adapt to near future. We tested the hypothesis that in ectotherms where reproduction is highly energy consuming, energy expenditure should vary throughout the breeding season with a maximum around nesting events. To test this hypothesis, we assessed the fine-scale behaviour, time-budget and estimated energetic costs in eight adult female European pond turtles Emys orbicularis equipped with data-loggers recording ambient temperature, pressure, light and the animals' 3-axis acceleration. Deployments occurred over four months throughout the nesting season 2017 in semi-natural captive conditions in Alsace, France. All study turtles showed a clear daily pattern over the 24h cycle, with four distinct phases (referred to as Night, Morning, Midday and Evening), associated with different behaviours and activity levels. Before oviposition, turtles were mostly active during Morning, and activity was positively driven by ambient temperature. Activity levels doubled during the nesting period, mostly due to the increased activity in the Evening, when nesting events occurred. Throughout the active season, basking occurrence at Midday was related to air temperature but cloud coverage was an even more important factor. Our results are a first step in predicting the seasonal time and energy budgets of the European pond turtle, and demonstrate the usefulness of animal-borne accelerometers to study free living freshwater turtles over extended periods of time. [ABSTRACT FROM AUTHOR]

Baseline plasma electrophoresis profiles (EPH) are important components of overall health and may aid in the conservation and captive management of species. The aim of this study was to establish plasma protein fractions for free-ranging Blanding's turtles (Emydoidea blandingii) and evaluate differences due to age class (adult vs. sub-adult vs. juvenile), sex (male, female, or unknown), year (2018 vs. 2019), month (May vs. June vs. July), health status, and geographical location (managed vs. unmanaged sites). Blood samples were obtained from 156 Blanding's turtles in the summer of 2018 and 129 in 2019 at two adjacent sites in Illinois. Results of the multivariate analysis demonstrated that age class, sex, year, month, health status, and geographical location all contributed to the variation observed in free-ranging populations. Adult females had the highest concentration of many protein fractions, likely associated with reproductive activity. Juveniles had lower protein concentrations. Temperature and rainfall differences between years impacted concentrations between 2018 and 2019, while May and June of both years saw higher levels in some protein fractions likely due to peak breeding and nesting season. Individuals with evidence of trauma or disease also showed increased plasma protein fractions when compared to those that were considered healthy. The two sites showed a wide/large variation over the two years. All of these factors emphasize the importance of considering multiple demographic or environmental factors when interpreting the EPH fractions. Establishing ranges for these analytes will allow investigation into disease prevalence and other environmental factors impacting this endangered species. [ABSTRACT FROM AUTHOR]

Late Jurassic deposits across Europe have yielded a rich fauna of extinct turtles. Although many of these turtles are recovered from marine deposits, it is unclear which of these taxa are habitually marine and which may be riverine species washed into nearby basins, as adaptations to open marine conditions are yet to be found. Two new fossils from the Late Jurassic of Germany provide unusually strong evidence for open marine adaptations. The first specimen is a partial shell and articulated hind limb from the Late Jurassic (early Tithonian) platy limestones of Schernfeld near Eichstätt, which preserves the integument of the hind limb as an imprint. The skin is fully covered by flat, polygonal scales, which stiffen the pes into a paddle. Although taxonomic attribution is not possible, similarities are apparent with Thalassemys. The second specimen is a large, articulated skeleton with hypertrophied limbs referable to Thalassemys bruntrutana from the Late Jurassic (early Late Kimmeridgian) platy limestone of Wattendorf, near Bamberg. Even though the skin is preserved as a phosphatic film, the scales are not preserved. This specimen can nevertheless be inferred to have had paddles stiffened by scales based on the pose in which they are preserved, the presence of epibionts between the digits, and by full morphological correspondence to the specimen from Schernfeld. An analysis of scalation in extant turtles demonstrated that elongate flippers stiffed by scales are a marine adaptation, in contrast to the elongate but flexible flippers of riverine turtles. Phylogenetic analysis suggests that Thalassemys bruntrutana is referable to the mostly Late Jurassic turtle clade Thalassochelydia. The marine adapted flippers of this taxon therefore evolved convergently with those of later clades of marine turtles. Although thalassochelydian fossils are restricted to Europe, with one notable exception from Argentina, their open marine adaptations combined with the interconnectivity of Jurassic oceans predict that the clade must have been even more wide-spread during that time. [ABSTRACT FROM AUTHOR]

Microbial communities of animals play a role in health and disease, including immunocompromised conditions. In the northeastern United States, cold-stunning events often cause endangered Kemp's ridley turtles (Lepidochelys kempii) to become stranded on beaches in autumn. These sea turtles are admitted to rehabilitation facilities when rescued alive and are presumed immunocompromised secondary to hypothermia. To better understand the role that microbes play in the health of cold-stunned sea turtles, we characterized the oral and cloacal microbiome from Kemp's ridley turtles at multiple timepoints during rehabilitation, from admission to pre-release, by using Illumina sequencing to analyze the 16S rRNA gene. Microbial communities were distinct between body sites and among turtles that survived and those that died. We found that clinical parameters such as presence of pneumonia or values for various blood analytes did not correlate with oral or cloacal microbial community composition. We also investigated the effect of antibiotics on the microbiome during rehabilitation and prior to release and found that the type of antibiotic altered the microbial community composition, yet overall taxonomic diversity remained the same. The microbiome of cold-stunned Kemp's ridley turtles gradually changed through the course of rehabilitation with environment, antibiotics, and disease status all playing a role in those changes and ultimately the release status of the turtles. [ABSTRACT FROM AUTHOR]

Clinical assessment of body condition is crucial in captive and free-ranging reptiles, since a large percentage of diseases result from inadequate nutrition. However, preventive health care is restricted by the lack of a practical method for the assessment in tortoises. Pre-existing evaluation systems based on weight and shell measurements are laborious and ignore the clinical presentation of the animal. The present study aimed to facilitate the assessment by establishing a body condition score. A total of 373 Hermann's Tortoises (Testudo hermanni) (n = 281 tortoises kept as pets in Germany and n = 92 tortoises originating from a free-ranging population (68) or a rearing station (24) in France) were examined and data (weight (g), carapace length (cm), width (cm), height (cm)) were recorded in a standard protocol between October 2020 and October 2021. A modified version of a body condition score for Mojave Desert Tortoises (Gopherus agassizii) (1 = cachectic, 3 = ideal, 5 = obese) was utilized and tested against pre-existing shell measurement systems (Jackson's ratio, body condition index, volume condition index, circumferential product). German captive tortoises were significantly heavier and larger than French specimens. In the Spearman's correlation matrix, the body condition score showed a statistically significant correlation with all measurement methods in the total population of captive tortoises (Testudo hermanni boettgeri), with a medium correlation strength, and a lack of correlation in free-ranging tortoises (Testudo hermanni hermanni). However, individual animal data suggested misleading results of mathematical equations in terms of body condition. Clinical evaluation of tortoises, including a body condition score, should be considered essential to provide good healthcare and should be an integral part of general examination., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Frankenberger et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

The variation in temperament among animals has consequences for evolution and ecology. One of the primary effects of consistent behavioral differences is on reproduction. In chelonians some authors have focused on the study of temperament using different methods. In our research our first aim was i) establish a methodology to determine the degree of boldness among individuals Rhinoclemmys areolata. Our second aim was to ii) determine the role boldness plays during reproduction, with emphasis on courtship and copulation, considering a) the interactions between males and females, and b) competition between males. We used 16 sexually mature individuals of each sex. Males were observed in four different situations and 17 behavioral traits were recorded. We selected 12 traits that allowed us distinguish between the bolder and the shier individuals and found that five behavioral traits were specific for bolder individuals and five others for shier individuals. In a second step, we observed a male in presence of a female and recorded courtship behaviors and breeding attempts. Bolder individuals did not display courtship behaviors and just attempted to copulate. Shier individuals displayed courtship behaviors and copulation attempts were rarely observed. Finally, in the simulations that compared two males in the presence of a female we noticed that bolder individuals displayed courtship behaviors while the shier ones simply ignored the female. Our results first allowed us to determine which methodology is the best to determine temperament in turtles. Secondly, temperament seems to be an important factor in modulating interaction between males and females. Bolder individuals have an advantage during competition and display courtship behaviours only if other males are present. Shier males displayed courtship behaviors and only try to copulate when no competitors were present. These two different temperament-dependant strategies are discussed in terms of ecology, evolution and management. [ABSTRACT FROM AUTHOR]

Microbial communities associated with freshwater aquatic habitats and resident species are both critical to and indicative of ecosystem status and organismal health. External surfaces of turtle shells readily accumulate microbial growth and could carry representation of habitat-wide microbial diversity, since they are in regular contact with multiple elements of freshwater environments. Yet, microbial diversity residing on freshwater turtle shells is poorly understood. We applied 16S and 18S metabarcoding to characterize microbiota associated with external shell surfaces of 20 red-eared slider (Trachemys scripta) turtles collected from varied habitats in central and western Oklahoma, and ranging to southeast Iowa. Shell-associated microbial communities were highly diverse, with samples dominated by Bacteroidia and alpha-/gamma-proteobacteria, and ciliophoran alveolates. Alpha diversity was lower on turtle shells compared to shallow-water-associated environmental samples, likely resulting from basking-drying behavior and seasonal scute shedding, while alpha diversity was higher on carapace than plastron surfaces. Beta diversity of turtle shells was similarly differentiated from environmental samples, although sampling site was consistently a significant factor. Deinococcus-Thermus bacteria and ciliophoran alveolates were recovered with significantly higher abundance on turtle shells versus environmental samples, while bacterial taxa known to include human-pathogenic species were variably more abundant between shell and environmental samples. Microbial communities from a single, shared-site collection of the ecologically similar river cooter (P. concinna) largely overlapped with those of T. scripta. These data add to a foundation for further characterization of turtle shell microbial communities across species and habitats, with implications for freshwater habitat assessment, microbial ecology and wildlife conservation efforts. [ABSTRACT FROM AUTHOR]

Blood was collected from wild captured green and Kemp's ridley turtles off the west coast of Florida, USA. Blood gases and biochemical values were analyzed using a point of care (POC) device in the field. Analytes include pH, partial pressure of carbon dioxide (pCO2), partial pressure of oxygen (pO2), total carbon dioxide (TCO2), bicarbonate (HCO3), base excess (BE), oxygen saturation (sO2), lactate, sodium (Na), potassium (K), chloride (Cl), total carbon dioxide (TCO2), anion gap, ionized calcium, glucose, blood urea nitrogen (BUN), creatinine (Crea), hematocrit (Hct), and hemoglobin (Hb). These are novel data for wild healthy Kemp's ridley turtles, and results for green turtles were generally consistent with past studies of green turtles with exceptions primarily in blood gas values. Ninety percent of the green turtles had fibropapillomatosis (FP), but none of the blood analytes were correlated with disease severity. Only BUN was correlated with weight of green turtles, and there was no correlation between blood parameters and weight of Kemp's ridley turtles. This study provides data that are useful in understanding the physiologic status of sea turtles specific to this region, allowing for comparisons to other populations, life stages, and disease states. [ABSTRACT FROM AUTHOR]

The acute phase response is a highly conserved reaction to infection, inflammation, trauma, stress, and neoplasia. Acute phase assays are useful for wildlife health assessment, however, they are infrequently utilized in reptiles. This study evaluated erythrocyte sedimentation rate (ESR) in eastern (Terrapene carolina carolina) and ornate box turtles (Terrapene ornata ornata) and hemoglobin-binding protein (HBP) in T. ornata. Erythrocyte sedimentation rate in 90 T. carolina and 105 T. ornata was negatively associated with packed cell volume and was greater in unhealthy turtles (p < 0.05). Female T. ornata had higher ESR values than males (p < 0.05). Measurement of ESR with a microhematocrit tube proportionally overestimated values from a commercial kit (Winpette), though both methods may retain utility with separate reference intervals. Hemoglobin-binding protein concentration in 184 T. ornata was significantly increased in adults and unhealthy turtles (p < 0.05). Erythrocyte sedimentation rate values were similar between seasons and populations, and HBP values were consistent between years, indicating that these analytes may have more stable baseline values than traditional health metrics in reptiles. This study demonstrates that ESR and HBP are promising diagnostics for health assessment in wild box turtles. Incorporating these tests into wild herptile health assessment protocols may support conservation efforts and improve ecosystem health monitoring. [ABSTRACT FROM AUTHOR]

The newly discovered plattenkalk (platy limestone) locality of Wattendorf, southern Germany, has yielded a diverse fauna and flora dated to the base of the late Kimmeridgian, Late Jurassic. We here describe three fossil turtle specimens that were recovered during systematic excavations of a distinct, 15 cm thick package of plattenkalks by the Naturkunde-Museum Bamberg. The first specimen is a large shell of Achelonia formosa, a taxon that is based on material from the late Kimmeridgian of Cerin, France. The new specimen suggests synonymy with Enaliochelys chelonia from the late Kimmeridgian of the United Kingdom. The second is a near-complete skeleton of the enigmatic Tropidemys seebachi, which was previously known only from the late Kimmeridgian of Hannover, northern Germany. The third specimen is a partial skeleton of Eurysternum wagleri, which had previously been known only from the early Tithonian of the Solnhofen region, southern Germany. In addition to new anatomical insights, the new material provides further evidence for spatial links during the late Kimmeridgian between northern and southern Germany, France, and the United Kingdom and temporal link from the late Kimmeridgian to the early Tithonian. The prevalence of partial, though articulated specimens is suggestive of predation by an unknown large marine reptile. [ABSTRACT FROM AUTHOR]

The Assam Roofed Turtle, Pangshura sylhetensis is an endangered and least studied species endemic to India and Bangladesh. The present study decodes the first complete mitochondrial genome of P. sylhetensis (16,568 bp) by using next-generation sequencing. The assembly encodes 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs), and one control region (CR). Most of the genes were encoded on the majority strand, except NADH dehydrogenase subunit 6 (nad6) and eight tRNAs. All PCGs start with an ATG initiation codon, except for Cytochrome oxidase subunit 1 (cox1) and NADH dehydrogenase subunit 5 (nad5), which both start with GTG codon. The study also found the typical cloverleaf secondary structures in most of the predicted tRNA structures, except for serine (trnS1) which lacks of conventional DHU arm and loop. Both Bayesian and maximum-likelihood phylogenetic inference using 13 concatenated PCGs demonstrated strong support for the monophyly of all 52 Testudines species within their respective families and revealed Batagur trivittata as the nearest neighbor of P. sylhetensis. The mitogenomic phylogeny with other amniotes is congruent with previous research, supporting the sister relationship of Testudines and Archosaurians (birds and crocodilians). Additionally, the mitochondrial Gene Order (GO) analysis indicated plesiomorphy with the typical vertebrate GO in most of the Testudines species. [ABSTRACT FROM AUTHOR]

Background: Conservation strategies are urgently needed for tropical turtles that are increasingly threatened by unsustainable exploitation. Studies conducted exclusively in temperate zones have revealed that typical turtle life history traits (including delayed sexual maturity and high adult survivorship) make sustainable harvest programs an unviable strategy for turtle conservation. However, most turtles are tropical in distribution and the tropics have higher, more constant and more extended ambient temperature regimes that, in general, are more favorable for population growth. Methods: To estimate the capacity of temperate and tropical turtles to sustain harvest, we synthesized life-history traits from 165 predominantly freshwater turtle species in 12 families (Carettochelydae, Chelidae, Chelydridae, Dermatemydidae, Emydidae, Geoemydidae, Kinosternidae, Pelomedusidae, Platysternidae, Podocnemididae, Staurotypidae and Trionychidae). The influence of climate variables and latitude on turtle life-history traits (clutch size, clutch frequency, age at sexual maturity, and annual adult survival) were examined using Generalized Additive Models. The biological feasibility of sustainable harvest in temperate and tropical species was evaluated using a sensitivity analysis of population growth rates obtained from stage-structured matrix population models. Results: Turtles at low latitudes (tropical zones) exhibit smaller clutch sizes, higher clutch frequency, and earlier age at sexual maturity than those at high latitudes (temperate zones). Adult survival increased weakly with latitude and declined significantly with increasing bioclimatic temperature (mean temperature of warmest quarter). A modeling synthesis of these data indicates that the interplay of life-history traits does not create higher harvest opportunity in adults of tropical species. Yet, we found potential for sustainable exploitation of eggs in tropical species. Conclusions: Sustainable harvest as a conservation strategy for tropical turtles appears to be as biologically problematic as in temperature zones and likely only possible if the focus is on limited harvest of eggs. Further studies are urgently needed to understand how the predicted population surplus in early life stages can be most effectively incorporated into conservation programs for tropical turtles. [ABSTRACT FROM AUTHOR]

Blood gas analysis reflects the exchange of oxygen and carbon dioxide in the lungs. This test provides important information, since the relationship between these gases has a direct impact on the acid-basic balance in the body. Given the significance of blood gas analysis in Brazilian reptiles, this study set out to establish temperature-corrected and uncorrected reference intervals for venous blood gas measurements in Chelonoidis carbonarius, and to compare values between females and males. In this study, 19 animals were used, 8 males and 11 females. Blood samples were collected from the dorsal coccygeal vein, and the analyses were performed immediately after blood sample collection. The following parameters were measured: pH, PO2, HCO3-, TCO2, BEecf, Na, K, ICa, and Glu, and were compared between females and males. Additionally, pH, pCO2, and pO2 values were compared with and without temperature correction. Oxygen saturation and Na levels were significantly higher (p<0.05) in males. Furthermore, it was possible to infer that the lower the body temperature relative to the environmental temperature, the larger the difference in pH following temperature correction., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2024 Silva La Rocca de Freitas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Kemp's ridley sea turtles were on the verge of extinction in the 1960s. While these sea turtles have slowly recovered, they are still critically endangered. In the last few years, the number of strandings on the beaches of Cape Cod, Massachusetts has increased by nearly an order of magnitude relative to preceding decades. This study uses a combination of ocean observations and a well-respected ocean model to investigate the causes and transport of cold-stunned sea turtles in Cape Cod Bay. After validating the model using satellite-tracked drifters and local temperature moorings, ocean currents were examined in Cape Cod Bay in an attempt to explain stranding locations as observed by volunteers and, for some years, backtracking was conducted to examine the potential source regions. The general finding of this study is that sub 10.5°C water temperatures in combination with persistently strong wind stress (>0.4 Pa), results in increased strandings along particular sections of the coast and are dependent on the wind direction. However, it is still uncertain where in the water column the majority of cold-stunned turtles reside and, if many of them are on the surface, considerable work will need to be done to incorporate the direct effects of wind and waves on the advective processes. [ABSTRACT FROM AUTHOR]

Chelonians are one of the most imperiled vertebrate taxa on the planet due to changes in the environment, anthropogenic influences, and disease. Over the last two decades, conservation strategies including nest protection, head-starting and meso-predator control have been successfully adopted by the Lake County Forest Preserve District for a population of state-endangered Blanding's turtles (Emydoidea blandingii) in Illinois. Only recently have efforts expanded to assess the effects of management action on turtle health. The objectives of this study were to 1) establish reference intervals for 16 hematologic and plasma biochemical analytes in free-ranging Blanding's turtles, 2) characterize demographic and temporal drivers of clinical pathology values including age class, sex, month, and year, and 3) describe bloodwork differences between a managed (SBCP) and unmanaged (IBSP) study site. Hematology and plasma biochemistries were performed for 393 turtles from 2017–18 at two sites in the Lake Plain region. Subject or population-based reference intervals were established based on the index of individuality per American Society for Veterinary Clinical Pathology guidelines. Analytes differed by age class [packed cell volume (PCV), total solids (TS), total white blood cell counts (WBC), heterophils, lymphocytes, heterophil:lymphocyte ratio (H:L), total calcium (Ca), calcium:phosphorous (Ca:P), bile acids (BA), aspartate aminotransferase (AST)], sex [H:L, Ca, phosphorus (P), Ca:P, creatine kinase (CK)], month [eosinophils, H:L, Ca, P, uric acid (UA), AST], and year [PCV, WBC, lymphocytes, basophils, H:L, Ca, P, UA]. Several analytes also varied by site [PCV, TS, monocytes, eosinophils, P, UA, AST], suggesting that health status may be affected by habitat management or lack thereof. The results of this study provide a baseline for ongoing health assessments in this region as well as across the Blanding's turtle range. [ABSTRACT FROM AUTHOR]

Bog turtles (Glyptemys muhlenbergii) are listed as Species of Greatest Conservation Need (SGCN) for wildlife action plans in every state it occurs and multi-state efforts are underway to better characterize extant populations and prioritize restoration efforts. However, traditional sampling methods can be ineffective due to the turtle's wetland habitat, small size, and burrowing nature. Molecular methods, such as qPCR, provide the ability to overcome this challenge by effectively quantifying minute amounts of turtle DNA left behind in its environment (eDNA). Developing such methods for bog turtles has proved difficult partly because of the high sequence similarity between bog turtles and closely-related, cohabitating species, most often wood turtles (Glyptemys insculpta). Additionally, substrates containing bog turtle eDNA are often rich in organics or other substances that frequently inhibit both DNA extraction and qPCR amplification. Here, we describe the development and validation of a qPCR assay, BT3, targeting the mitochondrial cytochrome oxidase I gene that correctly identifies bog turtles with 100% specificity and sensitivity when tested on 201 blood samples collected from six species over a wide geographic range. We also developed a full-process internal control employing a genetically modified strain of Caenorhabditis elegans to improve DNA extraction methods, limit false negative results due to qPCR inhibition, and measure total DNA recovery from each sample. Using the internal control, we found that DNA recovery varied by over an order of magnitude between samples and likely explains the lack of bog turtle detection in some cases. Methods presented herein are highly-specific and may offer a more cost effective, non-invasive tool to supplement bog turtle population assessments in the Eastern United States. Poor or differential DNA recovery, which remains unmeasured in the vast majority of eDNA studies, significantly reduced the ability to detect bog turtle in their natural environment. [ABSTRACT FROM AUTHOR]

The green turtle (Chelonia mydas) is a globally distributed marine species whose evolutionary history has been molded by geological events and oceanographic and climate changes. Divergence between Atlantic and Pacific clades has been associated with the uplift of the Panama Isthmus, and inside the Pacific region, a biogeographic barrier located west of Hawaii has restricted the gene flow between Central/Eastern and Western Pacific populations. We investigated the carapace shape of C. mydas from individuals of Atlantic, Eastern Pacific, and Western Pacific genetic lineages using geometric morphometrics to evaluate congruence between external morphology and species’ phylogeography. Furthermore, we assessed the variation of carapace shape according to foraging grounds. Three morphologically distinctive groups were observed which aligned with predictions based on the species’ lineages, suggesting a substantial genetic influence on carapace shape. Based on the relationship between this trait and genetic lineages, we propose the existence of at least three distinct morphotypes of C. mydas. Well-defined groups in some foraging grounds (Galapagos, Costa Rica and New Zealand) may suggest that ecological or environmental conditions in these sites could also be influencing carapace shape in C. mydas. Geometric morphometrics is a suitable tool to differentiate genetic lineages in this cosmopolitan marine species. Consequently, this study opens new possibilities to explore and test ecological and evolutionary hypotheses in species with wide morphological variation and broad geographic distribution range. [ABSTRACT FROM AUTHOR]

There is a vivid research on securely deliver a secret message by using a data hiding technique in digital images. However, most existing solutions of data-hiding need to encrypt secret data first, and embed the encrypt message into cover image, which is not promising any security once the embedding algorithm is leaked. In this paper, an efficient and adaptive data hiding scheme is presented which is based on a new reference matrix named secure reference matrix. The scheme is flexible enough to meet different data hiding capacities and image quality as needed, and we allow the reference matrix to be more secure since it is randomly generated each time and make it much harder for being attack with exhaustive manner by extending the possible solution. We give the detail protocol for our scheme and provide the practical test and performance analysis. Experimental results show that our scheme has a better flexibility and efficiency in embedding process, and the average value of Peak Signal to Noise Ratio (PSNR) for the stego-images which are generated by our scheme have a higher visual quality for different embedding capacities, and our scheme is more efficient compared with related work. [ABSTRACT FROM AUTHOR]

Sharks are top predators across ocean food webs and have a major ecological role in marine ecosystems. Investigating the trophic ecology of this group of species is thus essential to understand ecosystem functioning and inform specific management actions aimed at shark conservation. The Galapagos Islands represent one of the last ocean wildernesses, where populations of sharks and other top marine predators come close to a pristine status. Here we provide the first study on the trophic ecology of the tiger shark (Galeocerdo cuvier) within the Galapagos Marine Reserve (GMR), using a combination of stable isotope analysis, satellite tracking, and passive acoustic telemetry to investigate ontogenetic and spatial variations at two regions. The mean estimated δ13C and δ15N at Isabela island (western region) were -13.9 ± 0.5‰ and 13.7 ± 0.7‰; and for Santa Cruz island (central region) were -13.8 ± 0.3‰ and 13.4 ± 0.7‰, respectively. Green sea turtles (Chelonia mydas) were the main prey item for large tiger sharks (>280 cm TL), while smaller sharks mainly fed on squid and pelagic fish. Tiger sharks exhibited a high degree of philopatry around green sea-turtle nesting areas, with the majority of sharks detected around green sea-turtle nesting areas for at least 10 months after their capture date, and some individuals were even present during the entire three-year study period. Although we did not report statistically significant differences between the two regions, isotopic and electronic tagging data suggest that tiger sharks in the Galapagos could be segregated into specific populations separated by geographical scales of <100 km. The high productivity of the archipelago, along with the protection from industrial fishing granted by the GMR, result in abundant and predictable sources of prey. This high food abundance, combined with the presence of suitable habitats throughout the tiger shark life cycle, might result in a reduction of migratory behaviours when compared to movement patterns of tiger sharks in other ocean basins. Additional studies using genetic tools could provide further evidence on the presence of separate management units, as it has been recently revealed for other shark species inhabiting the GMR. [ABSTRACT FROM AUTHOR]

Plasma chemistry is widely used in diagnostic and research settings in sea turtles. However, plasma discolorations such as hemolysis are often not considered in data interpretation. The objectives of this study were to (1) evaluate the effects of moderate hemolysis on plasma electrolytes, minerals, and proteins using dry chemistry analysis (DCA) and protein electrophoresis from nesting leatherback sea turtles (Dermochelys coriacea) from Florida and to (2) establish blood analyte reference intervals. Twenty-six plasma samples with absence of hemolysis were selected and sub-divided into one non-hemolytic aliquot and an aliquot that was experimentally manipulated to mimic moderate hemolysis. Plasma samples were analyzed for hemoglobin using a handheld photometer; sodium, potassium, chloride, magnesium, calcium, phosphorus, and total protein using DCA; and protein electrophoresis. Packed cell volume and hemoglobin were measured in corresponding whole blood samples. Reference intervals were established. All analytes except calcium and pre-albumin were significantly higher and the calcium:phosphorus and albumin:globulin ratios were significantly lower in hemolytic plasma compared to non-hemolytic plasma. Alpha2-globulins and potassium were the analytes most impacted by hemolysis, averaging 3.3- and 2.0-fold higher in hemolyzed samples, respectively, indicating that (1) hemoglobin migrates into the alpha2-globulin region in this species and (2) notable intracellular potassium is released into plasma with hemolysis. Attempted conversion factors for compensation of hemolysis were considered inaccurate for 4 of 16 analytes due to non-significant regression lines. We also report that PCV provides an estimate of hemoglobin (g/L) using the formula: (2.59 × PCV) + 24.59. Given the spurious effects of hemolysis, the degree of this artifact should be reported with biochemistry data, and samples with moderate to severe hemolysis should be excluded from datasets when interpreting electrolyte, mineral, and protein results. This will ensure accurate data interpretation for individual turtles in rehabilitation or research settings and population-level data relevant to conservation-focused projects. [ABSTRACT FROM AUTHOR]

There is a wealth of published information on the epibiont communities of sea turtles, yet many of these studies have exclusively sampled epibionts found only on the carapace. Considering that epibionts may be found on almost all body-surfaces and that it is highly plausible to expect different regions of the body to host distinct epibiont taxa, there is a need for quantitative information on the spatial variation of epibiont communities on turtles. To achieve this, we measured how total epibiont abundance and biomass on olive ridley turtles Lepidochelys olivacea varies among four body-areas of the hosts (n = 30). We showed that epibiont loads on olive ridleys are higher, both in terms of number and biomass, on the skin than they are on the carapace or plastron. This contrasts with previous findings for other hard-shelled sea turtles, where epibionts are usually more abundant on the carapace or plastron. Moreover, the arguably most ubiquitous epibiont taxon for other hard-shelled sea turtles, the barnacle Chelonibia spp., only occurred in relatively low numbers on olive ridleys and the barnacles Stomatolepas elegans and Platylepas hexastylos are far more abundant. We postulate that these differences between the epibiont communities of different sea turtle taxa could indicate that the carapaces of olive ridley turtles provide a more challenging substratum for epibionts than do the hard shells of other sea turtles. In addition, we conclude that it is important to conduct full body surveys when attempting to produce a holistic qualitative or quantitative characterization of the epibiont communities of sea turtles. [ABSTRACT FROM AUTHOR]

Aims: The aims of this study were: a) to describe the pathological and laboratory findings in a case series of stranding and mortality associated with ingestion of large amounts of sea urchins in loggerhead turtles (Caretta caretta), and b) to alert veterinarians and biologists involved in sea turtle conservation of this cause of stranding and/or death. Methods: The six loggerheads studied were stranded on the coasts of Gran Canaria, Canary Islands, Spain, between 2008 and 2015. Post mortem studies included pathological, microbiological, and sea urchin species identification procedures. Results: All turtles showed severe intestinal impaction caused by large amounts of sea urchins, mainly affecting the colon and the caudal half of the small intestine. Histologically, severe focal fibrinonecrotic enteritis was diagnosed in two turtles. In the remaining turtles, lesions ranged from mild desquamation of the intestinal epithelium to severe congestion of the blood vessels of lamina propria, submucosa, muscular and serosa, and edema. Vibrio sp. was isolated from the spleen and intestinal mucosa of a loggerhead in which focal fibrinonecrotic enteritis had been diagnosed. In five turtles, all the remains were fragments from long-spined sea urchins (Diadema africanum); the last turtle contained a mixture of long-spined sea urchin (90%) and purple sea urchin (Sphaerechinus granularis) (10%) remains. Conclusions: Although the prevalence of this cause of stranding was low (< 1.6%) compared to other mortality causes, continued overfishing and anthropogenic climate change could increase its incidence. Intestinal impaction with large amounts of sea urchins should be included in the differential diagnosis of gastrointestinal diseases in sea turtles, and the possible toxic effect of some sea urchin species on sea turtles should also be investigated. [ABSTRACT FROM AUTHOR]

Caretta caretta is the most common sea turtle species in the Mediterranean Sea. The species is threatened by anthropomorphic activity that causes thousands of deaths and hundreds of strandings along the Mediterranean coast. Stranded turtles are often cared for in rehabilitation centres until they recover or die. The objective of this study was to characterize the gut microbiome of nine sea turtles stranded along the Sicilian coast of the Mediterranean Sea using high-throughput sequencing analysis targeting V3–V4 regions of the bacterial 16S rRNA gene. Stool samples were collected from eight specimens hosted in the recovery centre after a few days of hospitalization (under 7) and from one hosted for many weeks (78 days). To better explore the role of bacterial communities in loggerhead sea turtles, we compared our data with published fecal microbiomes from specimens stranded along the Tuscan and Ligurian coast. Our results highlight that, despite the different origin, size and health conditions of the animals, Firmicutes, Bacteroidetes and Proteobacteria constitute the main components of the microbiota. This study widens our knowledge on the gut microbiome of sea turtles and could be helpful for the set up of rehabilitation therapies of stranded animals after recovery in specialized centres. [ABSTRACT FROM AUTHOR]

The relationship between dive behavior and oceanographic conditions is not well understood for marine predators, especially sea turtles. We tagged loggerhead turtles (Caretta caretta) with satellite-linked depth loggers in the Gulf of Mexico, where there is a minimal amount of dive data for this species. We tested for associations between four measurements of dive behavior (total daily dive frequency, frequency of dives to the bottom, frequency of long dives and time-at-depth) and both oceanographic conditions (sea surface temperature [SST], net primary productivity [NPP]) and behavioral mode (inter-nesting, migration, or foraging). From 2011–2013 we obtained 26 tracks from 25 adult female loggerheads tagged after nesting in the Gulf of Mexico. All turtles remained in the Gulf of Mexico and spent about 10% of their time at the surface (10% during inter-nesting, 14% during migration, 9% during foraging). Mean total dive frequency was 41.9 times per day. Most dives were ≤ 25 m and between 30–40 min. During inter-nesting and foraging, turtles dived to the bottom 95% of days. SST was an important explanatory variable for all dive patterns; higher SST was associated with more dives per day, more long dives and more dives to the seafloor. Increases in NPP were associated with more long dives and more dives to the bottom, while lower NPP resulted in an increased frequency of overall diving. Longer dives occurred more frequently during migration and a higher proportion of dives reached the seafloor during foraging when SST and NPP were higher. Our study stresses the importance of the interplay between SST and foraging resources for influencing dive behavior. [ABSTRACT FROM AUTHOR]

This study uses satellite telemetry to track post-nesting movements of endangered green turtles (Chelonia mydas) (n = 6) in the Gulf of Guinea. It identifies a migratory corridor linking breeding grounds of Atlantic green turtles nesting on Bioko Island, Equatorial Guinea, to foraging grounds in the coastal waters of Accra, Ghana. Track lengths of 20–198 days were analyzed, for a total of 536 movement days for the six turtles. Migratory pathways and foraging grounds were identified by applying a switching state space model to locational data, which provides daily position estimates to identify shifts between migrating and foraging behavior. Turtles exhibited a combination of coastal and oceanic migrations pathways that ranged from 957 km to 1,131 km. Of the six turtles, five completed their migration and maintained residency at the same foraging ground near the coastal waters of Accra, Ghana until transmission was lost. These five resident turtles inhabit heavily fished waters and are vulnerable to a variety of anthropogenic threats. The identification of these foraging grounds highlights the importance of these coastal waters for the protection of the endangered Atlantic green turtle. [ABSTRACT FROM AUTHOR]

Leptospira spp. are re-emerging zoonotic pathogens. Previous research has found that Blanding's turtles (Emydoidea blandingii) experimentally infected with Leptospira interrogans shed leptospires in their urine, suggesting that they could play a role in transmitting pathogen within an aquatic ecosystem. This study investigated whether a population of wild Blanding's turtles known to be exposed to Leptospira spp. actively shed the pathogen under natural conditions. Blood samples were collected for serologic testing and to assess the health of the turtles. Free catch urine was collected for polymerase chain reaction (PCR) testing. All turtles were seropositive for Leptospira spp. and 73.5% (25/34) of the urine samples were PCR positive. All animals appeared clinically healthy and showed no apparent signs of disease. This study confirms that wild Blanding's turtles can actively shed Leptospira spp. in their urine and suggests that they may play a role in the epidemiology of this disease in habitats in which they reside. [ABSTRACT FROM AUTHOR]

Reptiles are declining worldwide yet our understanding of their immune function lags far behind other taxa. The innate immune system is the primary mode of defense in reptiles, and the serum complement cascade is its major component. We assessed serum complement activity of plasma in two closely related aquatic turtle species, the common snapping turtle (CST; Chelydra serpentina) and alligator snapping turtle (AST; Macrochelys temminckii). We used a sheep red blood cell (SRBC) hemolysis assay to assess serum complement activity. Although the antibacterial activities of the plasma of these turtle species are similar, the hemolytic activity was much stronger in CST than AST. Treatment with inhibitors of the serum complement cascade indicated differences in the mechanisms of complement activation between the turtle species. We subjected plasma from both turtle species to mannan affinity chromatography and analyzed the eluate with SDS-PAGE, which revealed that plasma from the CSTs contained only small amounts of one C-type lectin protein while the AST plasma contained high concentrations of two C-type lectins (31.0 and 35.9 kDa). Edman degradation analyses confirmed that the two AST proteins contained identical N-terminal sequences. Thus, the CST appears to rely more heavily on the alternative mechanism of serum complement activation, while the AST appears to rely more on the lectin-mediated pathway, which is a pattern recognition response to prokaryotes not activated by the SRBCs. These results are unique in that the use of serum complement pathways are generally assumed to be conserved within clades. [ABSTRACT FROM AUTHOR]