Crocodile Lizard Research Articles

Consumption of centipedes promotes growth and gut microbiome homeostasis in a lizard

AbstractAnimals are known for their self‐medicating behavior, and food composition also contributes to animal fitness. In this study, we investigated the dietary habits of endangered Chinese crocodile lizards and found that, compared to adult lizards, juvenile lizards consumed more centipedes, a trend validated through controlled feeding experiments. Juvenile lizards fed with centipedes demonstrated faster growth rates and better body conditions than those fed an earthworm‐only diet, despite no increase in food intake. The centipede‐added group showed lower species richness in the gut microbiome and had more abundant fermentative bacteria, such as Bacteroides, Paraclostridium, and Enterococcus, whereas the earthworm‐only group had more abundant non‐fermentative opportunistic pathogens. Higher concentrations of short‐chain fatty acids, which are mainly derived from bacterial fermentation in the gut, were found in the feces of the centipede‐added group. Therefore, consumption of centipedes promoted the growth and gut microbiota homeostasis of juvenile lizards. This study emphasizes the significance of preserving food diversity in natural environments and increasing dietary diversity in captive populations.

Outbreeding management offers the promise of genetic rescue for an endangered lizard

Abstract Inbreeding and loss of genetic diversity increase the extinction risk of small isolated populations. Genetic rescue by augmenting gene flow is a powerful means for the restoration of lost genetic variation. In this study, we used multigenerational pedigrees and neutral genetic markers to assess the consequences of outbreeding management in the Chinese and Vietnamese populations of the endangered crocodile lizard, Shinisaurus crocodilurus. Compared with the purebred Chinese population, the outbreeding population exhibited greater molecular genetic variation and a 3-fold larger population size. Moreover, the first-generation hybrids had a longer lifespan than purebreds, suggesting that outbreeding depression did not occur, but the long-term fitness effect of outbreeding needs to be further evaluated. Our study provides valuable insights into the potential for genetic rescue in the endangered crocodile lizard, emphasizing the importance of an evidence-based genetic management approach to address the risks of inbreeding and outbreeding depression in threatened populations.

Responses of gut microbiota in crocodile lizards (Shinisaurus crocodilurus) to changes in temperature.

The gut microbiota plays an essential role in maintaining the health and fitness of the host organism. As a critical environmental variable, temperature exerts significant effects on animal survival and reproduction. Elevated temperatures can influence the composition and function of the animal gut microbiota, which may have potentially detrimental effects on the host. The crocodile lizard (Shinisaurus crocodilurus) is an ancient and currently endangered reptile species due to human hunting and habitat destruction. Given the predicted shifts in global temperatures in the next century, it is important to understand how warming affects the gut microbiota of these vulnerable lizards, which remains unclear. To determine how the microbial communities change in crocodile lizards in response to warming, we analyzed the gut microbiota under five temperature conditions (22°C, 24°C, 26°C, 28°C, and 30°C) using 16S rRNA high-throughput sequencing. Results showed that the dominant phyla, Proteobacteria and Bacteroidetes, in gut microbiota were not significantly affected by temperature variations, but increasing temperature altered the structure and increased the community richness of the gut microbiota. In addition, warming changed the abundance of Pseudomonas aeruginosa and Actinobacteria, which may have negative effects on the physiological health of the crocodile lizards. Functional prediction analysis demonstrated that the functional pathways enriched in crocodile lizards were mainly related to metabolism, with no significant differences observed in these pathways at KEGG pathway level 1 after warming. These results provide valuable insights into the ecological adaptations and regulatory mechanisms employed by crocodile lizards in response to warming, which may be of benefit for their conservation.

It's a trap?! Escape from an ancient, ancestral sex chromosome system and implication of Foxl2 as the putative primary sex-determining gene in a lizard (Anguimorpha; Shinisauridae).

Although sex determination is ubiquitous in vertebrates, mechanisms of sex determination vary from environmentally to genetically influenced. In vertebrates, genetic sex determination is typically accomplished with sex chromosomes. Groups like mammals maintain conserved sex chromosome systems, while sex chromosomes in most vertebrate clades are not conserved across similar evolutionary timescales. One group inferred to have an evolutionarily stable mode of sex determination is Anguimorpha, a clade of charismatic taxa including monitor lizards, Gila monsters, and crocodile lizards. The common ancestor of extant anguimorphs possessed a ZW system that has been retained across the clade. However, the sex chromosome system in the endangered, monotypic family of crocodile lizards (Shinisauridae) has remained elusive. Here, we analyze genomic data to demonstrate that Shinisaurus has replaced the ancestral anguimorph ZW system on LG7 with a novel ZW system on LG3. The linkage group, LG3, corresponds to chromosome 9 in chicken, and this is the first documented use of this syntenic block as a sex chromosome in amniotes. Additionally, this ~1 Mb region harbors approximately 10 genes, including a duplication of the sex-determining transcription factor, Foxl2, critical for the determination and maintenance of sexual differentiation in vertebrates, and thus a putative primary sex-determining gene for Shinisaurus.

Structural variations in crocodile lizard populations provide novel insights into genomic variations in endangered animals.

Population-scale genome resequencing of endangered animals may contribute to gaining an understanding of how genomes vary as population sizes become smaller, as well as the functional implications of such variation. In this study, we analysed structural variations and gene presence and absence variations in the genomes of population of the endangered crocodile lizards. We found that the frequencies of some genes showed significant differences between crocodile lizards in different regions, indicating the influence of environmental selection, as well as potential contributions from demography and isolation, in shaping gene presence and absence variations. The haplotype diversity of major histocompatibility complex (MHC) genes was also found to differ among crocodile lizards inhabiting different regions. These findings indicate that well-designed interbreeding of crocodile lizards from different regions may facilitate the exchange of genes between different lizard populations and increase the haplotype diversity of MHC genes, which may be beneficial for the survival of these lizards. Our findings in this study, based on differences in gene structural variation, provide new insights into genomic variation and may contribute to the conservation of endangered animals.

Plastisphere characterization in habitat of the highly endangered Shinisaurus crocodilurus: Bacterial composition, assembly, function and the comparison with surrounding environment

Plastisphere is a new niche for microorganisms that complicate the ecological effects of plastics, and may profoundly influence biodiversity and habitat conservation. The DaGuishan National Nature Reserve, one of the largest habitats of the highly endangered crocodile lizard (Shinisaurus crocodilurus), is experiencing plastic pollution without sufficient attention. Here, plastisphere collected from captive tanks of crocodile lizards in this nature reserve was characterized for the first time. Three types of plastic (PE-PP, PE1, and PE2) together with the surrounding water and soil samples, were collected, and 16S rRNA sequencing technology was used to characterize the bacterial composition. The results demonstrated that plastisphere was driven by stochastic process and had a distinct bacterial community with higher diversity than that in surrounding water (p < 0.05). Bacteria related to nitrogen and carbon cycles (Pseudomonas psychrotolerans, Methylobacterium-Methylorubrum) were more abundant in plastisphere than in water or soil (p < 0.05). More importantly, plastics recruited pathogens and those bacteria function in antibiotic resistant genes (ARGs) coding. Bacteria related to polymer degradation also proliferated in plastisphere, especially Bacillus subtilis with a fold change of 42.01. The PE2 plastisphere, which had the lowest diversity and was dominated by Methylobacterium-Methylorubrum differed from PE 1 and PE-PP plastispheres totally. Plastics' morphology and aquatic nutrient levels contributed to the heterogeneity of different plastispheres. Overall, this study demonstrated that plastispheres diversify in composition and function, affecting ecosystem services directly or indirectly. Pathogens and bacteria related to ARGs expression enriched in the plastisphere should not be ignored because they may threaten the health of crocodile lizards by increasing the risk of infection. Plastic pollution control should be included in conservation efforts for crocodile lizards. This study provides new insights into the potential impacts of plastisphere, which is important for ecological risk assessments of plastic pollution in the habitats of endangered species.

Proteomic analysis of the mandibular glands from the Chinese crocodile lizard, Shinisaurus crocodilurus – Another venomous lizard?

Based on its phylogenetic relationship to monitor lizards (Varanidae), Gila monsters (Heloderma spp.), and the earless monitor Lanthanotus borneesis, the Chinese crocodile lizard, Shinisaurus crocodilurus, has been assigned to the Toxicofera clade, which comprises venomous reptiles. However, no data about composition and biological activities of its oral secretion have been reported. In the present study, a proteomic analysis of the mandibular gland of S. crocodilurus and, for comparison, of the herbivorous Solomon Island skink Corucia zebrata, was performed. Scanning electron microscopy (SEM) of the teeth from S. crocodilurus revealed a sharp ridge on the anterior surface, but no grooves, whereas those of C. zebrata possess a flattened crown with a pointed cusp. Proteomic analysis of their gland extracts provided no evidence of venom-derived peptides or proteins, strongly supporting the non-venomous character of these lizards. Data are available via ProteomeXchange with identifier PXD039424.

Genome of the endangered Guatemalan Beaded Lizard, Heloderma charlesbogerti, reveals evolutionary relationships of squamates and declines in effective population sizes.

Many lizard species face extinction due to worldwide climate change. The Guatemalan Beaded Lizard, Heloderma charlesbogerti, is a member of the Family Helodermatidae that may be particularly imperiled; fewer than 600 mature individuals are believed to persist in the wild. In addition, H. charlesbogerti lizards are phenotypically remarkable. They are large in size, charismatically patterned, and possess a venomous bite. Here, we report the draft genome of the Guatemalan Beaded Lizard using DNA from a wild-caught individual. The assembled genome totals 2.31 Gb in length, similar in size to the genomes of related species. Single-copy orthologs were used to produce a novel molecular phylogeny, revealing that the Guatemalan Beaded Lizard falls into a clade with the Asian Glass Lizard (Anguidae) and in close association with the Komodo Dragon (Varanidae) and the Chinese Crocodile Lizard (Shinisauridae). In addition, we identified 31,411 protein-coding genes within the genome. Of the genes identified, we found 504 that evolved with a differential constraint on the branch leading to the Guatemalan Beaded Lizard. Lastly, we identified a decline in the effective population size of the Guatemalan Beaded Lizard approximately 400,000 years ago, followed by a stabilization before starting to dwindle again 60,000 years ago. The results presented here provide important information regarding a highly endangered, venomous reptile that can be used in future conservation, functional genetic, and phylogenetic analyses.

Climate Change and Dispersal Ability Jointly Affects the Future Distribution of Crocodile Lizards.

Simple SummaryClimate change is one of the most significant global challenges we face this century, and all species will likely be affected by it. Climate change may cause organisms to advance their phenology, delay reproduction, shift their ranges northwards, or even reduce their range. Reptiles, as typical ectotherms, are more sensitive to climate change, mainly in the form of range shifts. The crocodile lizard (Shinisaurus crocodilurus Ahl, 1930) belongs to a monophyletic family and genus and is a relict species from the late Quaternary glacial period. Given its unique evolutionary history, it arguably has greater conservation value. The overall population size of this species is decreasing due to habitat destruction and over-exploitation (e.g., traditional medicine, food, and pet trade). This study evaluated the effect of climate change and dispersal capacity on the potential distribution range of crocodile lizards. Temperature and precipitation interact to affect the distribution of crocodile lizards. Historical climate has seen a large expansion in the range of the crocodile lizard. However, in a future global warming scenario, the potential range of crocodile lizards will continue to shrink. The same results were obtained for different migration scenarios. However, to better predict future changes to the distribution range of the crocodile lizard we need more fieldwork and ground truthing. All we can do currently is try to protect the lizard and its habitat to ensure their continued survival.Crocodile lizards (Shinisaurus crocodilurus) are an endangered, ‘living fossil’ reptile from a monophyletic family and therefore, a high priority for conservation. We constructed climatic models to evaluate the potential impact of climate change on the distribution of crocodile lizards for the period 2000 to 2100 and determined the key environmental factors that affect the dispersal of this endangered species. For the construction of climatic models, we used 985 presence-only data points and 6 predictor variables which showed excellent performance (AUC = 0.974). The three top-ranked factors predicting crocodile lizard distribution were precipitation of the wettest month (bio13, 37.1%), precipitation of the coldest quarter (bio19, 17.9%), and temperature seasonality (bio4, 14.3%). Crocodile lizards were, just as they are now, widely distributed in the north of Guangdong Province in China and Quảng Ninh Province in Vietnam at the last glacial maximum (LGM). Since the LGM, there has been an increase in suitable habitats, particularly in east-central Guangxi Province, China. Under future global warming scenarios, the potential habitat for crocodile lizards is expected to decrease significantly in the next 100 years. Under the most optimistic scenario, only 7.35% to 6.54% of suitable habitat will remain, and under the worst climatic scenario, only 8.34% to 0.86% of suitable habitat will remain. Models for no dispersal and limited dispersal showed that all crocodile lizards would lose habitat as temperatures increase. Our work contributes to an increased understanding of the current and future spatial distribution of the species, supporting practical management and conservation plans.

Investigation of Parasitic Infection in Crocodile Lizards (Shinisaurus crocodilurus) Using High-Throughput Sequencing.

Simple SummaryTo explore unexplained crocodile lizard deaths, we analyzed fecal samples from crocodile lizards at the Daguishan and Luokeng nature reserves using high-throughput sequencing to investigate parasitic infections in this endangered species. The present study reported the occurrence of parasitic infection in wild and captive crocodile lizards, with a total parasitic infection rate of 33.33% (23/69). Different influencing factors (populations and regions) were analyzed, and significant differences were found. The results indicate that captive crocodile lizards were more susceptible to parasitic diseases than wild crocodile lizards. In addition, only Cryptosporidium infection varied by geographical location, with crocodile lizards from Daguishan showing higher susceptibility to Cryptosporidium infestation. From a disease prevention perspective, these findings have great significance for crocodile lizard conservation.The highly endangered crocodile lizard (Shinisaurus crocodilurus) continues to be impacted by disease, especially in captive breeding populations. In this paper, based on high-throughput sequencing, we investigated parasitic infections in captive and wild crocodile lizard populations in the Daguishan National Nature Reserve and Guangdong Luokeng Shinisaurus crocodilurus National Nature Reserve. The results show that the overall parasitic infection rate in crocodile lizards was 33.33% (23/69). Four parasite genera were detected, including Eimeria, Cryptosporidium, Nematopsis, and Acanthamoeba, with infection rates of 15.94% (11/69), 17.39% (12/69), 7.25% (5/69), and 4.35% (3/69), respectively. Significant differences in the infection rate were found between the different parasite species (χ2 = 8.54, p < 0.05, chi-squared test). The parasitic infection rates in the captive and wild populations were 39.29% (22/56) and 7.69% (1/13), respectively, which were significantly different (p < 0.05, Fisher’s exact test). However, no significant differences in the infection rates of the four parasite genera were found between the captive and wild populations (p > 0.05, Fisher’s exact test). The parasitic infection rates in Daguishan and Luokeng were 34.09% (15/44) and 32.00% (8/25), respectively, which were not significantly different (p > 0.05, Fisher’s exact test). However, significant differences in terms of species were found in the two reserves (p < 0.01, Fisher’s exact test). Only Cryptosporidium infection showed a significant difference between the two regions (p < 0.01, Fisher’s exact test). Our results suggest that captive crocodile lizards are more susceptible to parasitic diseases than wild crocodile lizards and that Cryptosporidium infection varies by geographical region. This study provides basic information about the parasites of endangered crocodile lizards, as well as a reference for disease control and conservation.

Potential impact of land-use change on habitat quality in the distribution range of crocodile lizards in China.

The over‐exploitation of land resources poses a serious threat to biodiversity on a global scale. Changes in land‐use and human exploitation have had a major impact on wild populations and their habitat in China. We assessed how habitat quality has changed over time (1995–2020). Specifically, we analyzed how the habitat quality of crocodile lizard has changed over time based on multi‐temporal land‐use data (1995, 2000, 2010, 2015 and 2020) using a land‐use transfer matrix and habitat quality model. The results showed that the main landscape types in the study area were arable land (21.21% of the area) and woodland (69.59% of the area) during the period. Construction land (land used for development) had decreased by 991 km2, a decrease rate of 59.84% from 1995 to 2000, and increased to 2349 km2, an increase rate of 71.69% from 2000 to 2020. The proportion of grasslands and areas with water were negligible and overall, did not vary significantly in size over the study period. The main feature of land use change in the study area was the loss of grasslands and woodlands through development. The habitat quality model indicated that habitat quality was highest and degradation was lowest in Dayao mountain, Guxiu town, Qichong village and Beituo town. Habitat quality improved in Daguishan and Luokeng areas. Habitat quality was good in Daping mountain and Linzhouding, but they were highly fragmented with patches of low‐quality habitat of varying sizes. Habitats were severely degraded in the Dateng Gorge area. The rate of habitat degradation has slowed over time in the study area, but gradually increased in degradation intensity, and low‐quality habitats were widely distributed and overlapped with the crocodile lizards distribution area. We recommend that protected areas for the crocodile lizard be more closely monitored and managed to halt further decline in habitat quality.

The eggshell-matrix protein gene OC-17 is functionally lost in the viviparous Chinese crocodile lizard.

Thickness reduction or loss of the calcareous eggshell is one of major phenotypic changes in the transition from oviparity to viviparity. Whether the reduction of eggshells in viviparous squamates is associated with specific gene losses is unknown. Taking advantage of a newly generated high-quality genome of the viviparous Chinese crocodile lizard (Shinisaurus crocodilurus), we found that ovocleidin-17 gene (OC-17), which encodes an eggshell matrix protein that is essential for calcium deposition in eggshells, is not intact in the crocodile lizard genome. Only OC-17 transcript fragments were found in the oviduct transcriptome, and no OC-17 peptides were identified in the eggshell proteome of crocodile lizards. In contrast, OC-17 was present in the eggshells of the oviparous Mongolia racerunner (Eremias argus). Although the loss of OC-17 is not common in viviparous species, viviparous squamates show fewer intact eggshell-specific proteins than oviparous squamates. Our study implies that functional loss of eggshell-matrix protein genes may be involved in the reduction of eggshells during the transition from oviparity to viviparity in the crocodile lizard.

Colour Variation in the Crocodile Lizard (Shinisaurus crocodilurus) and Its Relationship to Individual Quality.

Simple SummaryThis study examines colour variation in the highly endangered crocodile lizard, Shinisaurus crocodilurus. Both males and females vary in the extent to which their throats and venters are red. Their colouration is easily visible to a lizard receiver, and we found evidence that colour signals individual quality. Females with red venters had larger heads while females with red throats had greater bite force. In males, redder individuals were older. Finally, we found links between colour and fitness in males but not females. Aspects of male colouration were linked to reproductive output such that they sired offspring from heavier litters. The potential fitness consequences of colour should be considered in captive breeding and release programs.Colour plays a key role in animal social communication including as an indicator of individual quality. Using spectrophotometry, we examined colour variation in the throat and venter of the crocodile lizard (Shinisaurus crocodilurus), an endangered species native to southern China and northern Vietnam. We detected two broad colour variants, individuals with and without red, for each body region and each sex. A cluster analysis of spectral colour measurements (hue, chroma, luminance) revealed discrete throat and ventral morphs when measured in a single snapshot in time. However, photographic evidence revealed that the amount of red relative to body size increased as they got older. Individuals with red were equally likely to be male or female and throat colour was unrelated to ventral colour. Therefore, it is premature to claim that crocodile lizards have discrete colour morphs. We used visual modelling to show that the throat and venter were easily discriminable to a lizard visual system, suggesting they function in social communication. We also asked whether colour variation signalled individual quality. Females with red throats had greater bite force while males with red throats were older. In addition, females with red venters had larger heads. We also detected differences in morphology linked to colour. Females with red throats had slender bodies and longer tails, while individuals lacking red on their throats were stouter and had shorter tails. Finally, throat and ventral colour were unrelated to reproductive output (litter size and mass) in females. Males with greater ventral luminance contrast sired offspring from litters with greater litter mass (including stillborns), while males with greater ventral chromatic contrast sired offspring whose collective live mass (excluding stillborns) was greater. Males with greater luminance contrast also sired more live offspring (excluding stillborns). Collectively, these results suggest that male ventral colour signals individual quality in males. Conservation initiatives should take colour variation into account when planning future captive breeding and release programs for this endangered species.

Will climatic changes affect the Vietnamese crocodile lizard? Seasonal variation in microclimate and activity pattern of Shinisaurus crocodilurus vietnamensis

Abstract Climate change is considered to negatively affect vertebrate biodiversity. Lizards in tropical regions are considered particularly vulnerable due to their narrow temperature tolerances. The crocodile lizard Shinisaurus crocodilurus, an ecologically specialized semiaquatic species, is under the risk of extinction due to habitat loss and overharvesting. It is only found in isolated relict populations in southern China and northern Vietnam. Data on the thermal niche of the species are crucial to assess its vulnerability to climatic changes. We developed a backpack system with temperature data loggers for S. crocodilurus in order to monitor the lizard’s temperature preferences in its natural habitat in Vietnam. We found that S. crocodilurus vietnamensis preferred a mean ambient temperature range of 24 ± 1°C (min-max: 22-31°C) at natural habitat sites, a far lower temperature amplitude compared to the recorded ambient temperature range at habitat sites. In its natural habitat, S. crocodilurus vietnamensis avoided in particular high temperatures, indicating the importance of constantly cool streams and intact vegetation for the provision of shade and shelter. We further provide first insights into the seasonal variation in microhabitat use and activity of S. crocodilurus vietnamensis based on data from individuals kept in outdoor enclosures under natural climatic conditions in Vietnam, collected daily over a period of eight months. Our results aim to aid further conservation measures for the species, such as the identification and protection of core habitat sites and sites for restocking, as well as to adjust and improve conservation breeding programs for the species.

Identification of Pseudomonas aeruginosa From the Skin Ulcer Disease of Crocodile Lizards (Shinisaurus crocodilurus) and Probiotics as the Control Measure.

The crocodile lizard (Shinisaurus crocodilurus) is an endangered ancient reptile species. Captive breeding is an important conservation measure for the potential restoration and recovery of their wild populations. However, a skin ulcer disease caused by an unknown pathogen has become a serious threat to captive breeding individuals. In the current study, based on microbial isolation, we identified Pseudomonas aeruginosa as the dominant pathogen in skin ulcer disease. Chinese skinks (Plestiodon chinensis) were used to verify the pathogenicity of P. aeruginosa in skin ulcer disease in vivo. As expected, subcutaneous inoculation of P. aeruginosa induced skin disease in healthy skinks and P. aeruginosa was re-isolated from the induced skin ulcers. Therefore, P. aeruginosa, an opportunistic and ubiquitous pathogen that causes a wide range of infections, appears to be the main pathogen of the skin disease affecting crocodile lizards. In the aquaculture industry, probiotics are widely used in the prevention and control of animal diseases caused by such pathogens. Here, we administered probiotics to the breeding crocodile lizards for 6 months. The three experiment groups treated with different kinds of probiotics showed significance at controlling case incidence. Three of the four groups treated with probiotics showed significant disease prevention (Effective Microorganisms mixed probiotics P = 0.0374; Double-dose Effective Microorganisms, P = 0.0299; Bacillus subtilis, P = 0.0140, T-test), and CFUs in the water of the breeding enclosures were also inhibited after probiotics usage (P < 0.001, T-test). Our study demonstrated the role of Pseudomonas aeruginosa in development of skin ulcer disease of crocodile lizards in a local zoo and offered the probiotic-based method for control measurements, which would be of benefit for the conservation of endangered reptiles.

Molecular phylogenetic analyses and ecological niche modeling provide new insights into threats to the endangered Crocodile Lizard (Shinisaurus crocodilurus)

The endangered crocodile lizard, Shinisaurus crocodilurus, is seriously imperiled by anthropogenic threats, including habitat loss and degradation and most critically over-collection for the international pet trade. As a result, population sizes of crocodile lizards have sharply declined throughout their range, with only a small number remaining in China and a handful of individuals left in Vietnam. To prioritize conservation measures for the species, in this study, we generate new mitochondrial sequences of important new samples and analyze them with existing data. Our results confirm a new genetically distinct population in China, highlighting cryptic genetic diversity within the species. The assessment of climate change impacts on the species suggests that the suitable habitat of one population in China will become severely fragmented, decreased in size, and shifted, while the habitat of two other Chinese populations will remain stable and may even expand given future climate scenarios. In Vietnam, Yen Tu Mountain Range is predicted to serve as a refuge in different climate change projections, but suitable habitat in the border area between Vietnam and China will likely shift geographically. Based on our results, we recommend further field surveys to locate the natural distribution range of the newly identified molecular clade in China and determine the distribution extension of the Vietnamese population in the border area, especially potential occurrence on the Chinese side. Considering the impacts of climate change on the Vietnamese population, designing a corridor to connect the subpopulation’s habitat in the border area with nature reserves in Yen Tu Mountain Range and/or translocating lizards from the site to more suitable habitats might help secure the subpopulation in the context of climate change. In all recommended conservation measures, close collaboration between Vietnam and China will be crucial to effectively protect this potentially shared subpopulation of the highly threatened species.

Skin Microbiota Was Altered in Crocodile Lizards (Shinisaurus crocodilurus) With Skin Ulcer.

Skin diseases commonly affect reptiles, but their relationships to the closely related skin microbiome are not well-understood. In recent years, both the wild and captive populations of the crocodile lizard, a Class I protected endangered animal in China, have suffered serious skin diseases that hamper the rescue and release projects for their conservation. This study conducted a detailed prevalence investigation of a major dermatosis characterized by foot skin ulcer in crocodile lizards. It should be noticed that skin ulcer has been prevalent in both captive and wild populations. There was positive correlation between skin ulcer and temperature, while no significant relationship between skin ulcer and humidity, sex, and age. We further studied the relationship between skin ulcer and the skin microbiota using meta-taxonomics. Results showed that the skin microbiota of crocodile lizards was significantly different from those of the environmental microbial communities, and that skin microbiota had a significant relationship with skin ulcer despite the impact of environment. Both bacterial and fungal communities on the ulcerated skin were significantly changed, which was characterized by lower community diversity and different dominant microbes. Our findings provide an insight into the relationship between skin microbiota and skin disease in reptile, serving as a reference for dermatological etiology in wildlife conservation.

Ancient Demographics Determine the Effectiveness of Genetic Purging in Endangered Lizards.

The purging of deleterious alleles has been hypothesized to mitigate inbreeding depression, but its effectiveness in endangered species remains debatable. To understand how deleterious alleles are purged during population contractions, we analyzed genomes of the endangered Chinese crocodile lizard (Shinisaurus crocodilurus), which is the only surviving species of its family and currently isolated into small populations. Population genomic analyses revealed four genetically distinct conservation units and sharp declines in both effective population size and genetic diversity. By comparing the relative genetic load across populations and conducting genomic simulations, we discovered that seriously deleterious alleles were effectively purged during population contractions in this relict species, although inbreeding generally enhanced the genetic burden. However, despite with the initial purging, our simulations also predicted that seriously deleterious alleles will gradually accumulate under prolonged bottlenecking. Therefore, we emphasize the importance of maintaining a minimum population capacity and increasing the functional genetic diversity in conservation efforts to preserve populations of the crocodile lizard and other endangered species.

Age-Dependent Enclosure Use in Juvenile Chinese Crocodile Lizards, Shinisaurus crocodilurus crocodilurus

This study compared the resource use of juvenile zoo-living Chinese crocodile lizards, Shinisaurus crocodilurus crocodilurus across three observation windows, spanning nine months, accounting for time of day and lizard age, and under consistent environmental conditions. Lizards showed a significant difference in proportionate resource use, quantified using a modified spread of participation indices between the second and final sampling period, such that with increasing age, resources were more equally utilised. The time of day did not have a significant effect on resource use. Lizards in this study significantly increased their use of water bodies and branches outside the bask zone and decreased their use of the land areas within the bask zones over time. Resource use data suggests the importance of providing enclosures which cater to ontogenetic shifts in captive individuals or within mixed age groupings.

Monitoring a loss: Detection of the semi‐aquatic crocodile lizard (Shinisaurus crocodilurus) in inaccessible habitats via environmental DNA

Assessing the conservation status of a species is strongly dependent upon data on species distribution and abundance. With the emergence of novel methods for species monitoring – such as the use of environmental DNA (eDNA) – monitoring success can be improved at reduced expenditure in the field, particularly in remote regions and terrains where access is difficult or dangerous. The highly endangered crocodile lizard (Shinisaurus crocodilurus Ahl, 1930) inhabits fragmented sites of the remaining evergreen forest with running water systems in a narrow distribution range in southern China and north‐east Vietnam. Crocodile lizards spend most of the day within or above water bodies, which are commonly remote and inaccessible. To monitor recent spatial occurrences, and to confirm the persistence or extinction of previously reported populations (especially in heavily altered habitats), the suitability of using eDNA and quantitative polymerase chain reaction (qPCR) was tested as an alternative method for monitoring this semiaquatic lizard. To assess the accuracy and limitations of this method, eDNA results from the field were compared with eDNA data from mesocosms and census data on the actual abundance of this species in the field. Environmental DNA of the crocodile lizard was detected in all of the positive controls, and in four of six natural sites; thus, all data collected using traditional field surveys were confirmed with eDNA results. eDNA monitoring was found to be a reliable method for assessing the viability of populations; we suggest that it should be developed as a tool for efficient wildlife management, particularly under difficult field and funding conditions.