Lizards and the enzootic cycle of Borrelia burgdorferi sensu lato.

Emerging and re-emerging pathogens often stem from zoonotic origins, cycling between humans and animals, and are frequently vectored and maintained by hematophagous arthropod vectors. The efficiency by which these disease agents are successfully transmitted between vertebrate hosts is influenced by many factors, including the host on which a vector feeds. The Lyme disease bacterium Borrelia burgdorferi sensu lato has adapted to survive in complex host environments, vectored by Ixodes ticks, and maintained in multiple vertebrate hosts. The versatility of Lyme borreliae in disparate host milieus is a compelling platform to investigate mechanisms dictating pathogen transmission through complex networks of vertebrates and ticks. Squamata, one of the most diverse clade of extant reptiles, is comprised primarily of lizards, many of which are readily fed upon by Ixodes ticks. Yet, lizards are one of the least studied taxa at risk of contributing to the transmission and life cycle maintenance of Lyme borreliae. In this review, we summarize the current evidence, spanning from field surveillance to laboratory infection studies, supporting their contributions to Lyme borreliae circulation. We also summarize the current understanding of divergent lizard immune responses that may explain the underlying molecular mechanisms to confer Lyme spirochete survival in vertebrate hosts. This review offers a critical perspective on potential enzootic cycles existing between lizard-tick-Borrelia interactions and highlights the importance of an eco-immunology lens for zoonotic pathogen transmission studies.

Seasonal dietary changes relate to gut microbiota composition depending on the host species but do not correlate with gut microbiota diversity in arthropod-eating lizards.

The animal gut microbiota is strongly influenced by environmental factors that shape their temporal dynamics. Although diet is recognized as a major driver of gut microbiota variation, dietary patterns have seldom been linked to gut microbiota dynamics in wild animals. Here, we analysed the gut microbiota variation between dry and rainy seasons across four Sceloporus species (S. aeneus, S. bicanthalis, S. grammicus and S. spinosus) from central Mexico in light of temporal changes in diet composition. The lizard microbiota was dominated by Firmicutes (now Bacillota) and Bacteroidota, and the closely related species S. aeneus and S. bicanthalis shared a great number of core bacterial taxa. We report species-specific seasonal changes in gut microbiota diversity and composition: greater alpha diversity during the dry compared to the rainy season in S. bicanthalis, the opposite pattern in S. aeneus, and no seasonal differences in S. grammicus and S. spinosus. Our findings indicated a positive association between gut bacterial composition and dietary composition for S. bicanthalis and S. grammicus, but bacterial diversity did not increase linearly with dietary richness in any lizard species. In addition, seasonality affected bacterial composition, and microbial community similarity increased between S. aeneus and S. bicanthalis, as well as between S. grammicus and S. spinosus. Together, our results illustrate that seasonal variation and dietary composition play a role in shaping gut microbiota in lizard populations, but this is not a rule and other ecological factors influence microbiota variation.

Bacterial pathogens in Ixodes ricinus collected from lizards Lacerta agilis and Zootoca vivipara in urban areas of Wroclaw, SW Poland- preliminary study.

The aim of this study was to determine the level of infection of Ixodes ricinus ticks with pathogens (Borrelia spp., Rickettsia spp., and Anaplasma spp.) collected from Lacerta agilis and Zootoca vivipara lizards in the urban areas of Wroclaw (SW Poland). The study was carried out in July-August 2020. Lizards were caught by a noose attached to a pole or by bare hands, identified by species, and examined for the presence of ticks. Each lizard was then released at the site of capture. Ticks were removed with tweezers, identified by species using keys, and molecular tests were performed for the presence of pathogens. From 28 lizards (17 specimens of Z. vivipara and 11 specimens of L. agilis) a total of 445 ticks, including 321 larvae and 124 nymphs, identified as I. ricinus were collected. A larger number of ticks were obtained from L. agilis compared to Z. vivipara. Molecular tests for the presence of pathogens were performed on 445 specimens of I. ricinus. The nested PCR method for the fla gene allowed the detection of Borrelia spp. in 9.4% of ticks, and it was higher in ticks from L. agilis (12.0%) than from Z. vivipara (1.0%). The RFLP method showed the presence of three species, including two belonging to the B. burgdorferi s.l. complex (B. lusitaniae and B. afzelii), and B. miyamotoi. The overall level of infection of Rickettsia spp. was 19.3%, including 27.2% in ticks collected from Z. vivipara and 17.0% from L. agilis. Sequencing of randomly selected samples confirmed the presence of R. helvetica. DNA of Anaplasma spp. was detected only in one pool of larvae collected from L. agilis, and sample sequencing confirmed the presence of (A) phagocytophilum. The research results indicate the important role of lizards as hosts of ticks and their role in maintaining pathogens in the environment including urban agglomeration as evidenced by the first recorded presence of (B) miyamotoi and (A) phagocytophilum in I. ricinus ticks collected from L. agilis. However, confirmation of the role of sand lizards in maintaining (B) miyamotoi and A. phagocytophilum requires more studies and sampling of lizard tissue.

Endoparasite Infections in Captive Inland Bearded Dragons (Pogona vitticeps) in Italy.

The inland bearded dragon (Pogona vitticeps) is a lizard species commonly kept as a pet worldwide. Endoparasites are among the most important pathogens affecting bearded dragons. The aim of this study was to evaluate the endoparasites in captive P. vitticeps in Italy. Faecal samples from 30 P. vitticeps were analysed by fresh faecal smears, flotation tests, the Mini-FLOTAC technique, and a rapid immunoassay to detect Cryptosporidium spp. To search for microsporidia, PCR and sequencing were performed on the faecal samples. Data were statistically analysed. The overall positivity rate for endoparasites was 83.3% (25/30). The identified endoparasites were oxyurids (17/30, 56.7%), Isosospora amphiboluri (13/30, 43.3%), Encephalitozoon pogonae (4/18, 22.22%), and Cryptosporidium sp. (1/30, 3.33%). The positivity for protozoa was significantly higher in juveniles compared to adults. Moreover, the frequency of clinical signs was significantly higher in the positive animals. The results obtained here emphasize the importance of regular veterinary examinations of captive P. vitticeps, aimed at the diagnosis, treatment, and control of endoparasites. This study is one of the largest surveys on microsporidia infections in living bearded dragons, suggesting that E. pogonae may be widespread in this lizard.

Fossil-informed biogeographic analysis suggests Eurasian regionalization in crown Squamata during the early Jurassic.

Background: Squamata (lizards, snakes, and amphisbaenians) is a Triassic lineage with an extensive and complex biogeographic history, yet no large-scale study has reconstructed the ancestral range of early squamate lineages. The fossil record indicates a broadly Pangaean distribution by the end- Cretaceous, though many lineages (e.g., Paramacellodidae, Mosasauria, Polyglyphanodontia) subsequently went extinct. Thus, the origin and occupancy of extant radiations is unclear and may have been localized within Pangaea to specific plates, with potential regionalization to distinct Laurasian and Gondwanan landmasses during the Mesozoic in some groups. Methods: We used recent tectonic models to code extant and fossil squamate distributions occurring on nine discrete plates for 9,755 species, with Jurassic and Cretaceous fossil constraints from three extinct lineages. We modeled ancestral ranges for crown Squamata from an extant-only molecular phylogeny using a suite of biogeographic models accommodating different evolutionary processes and fossil-based node constraints from known Jurassic and Cretaceous localities. We hypothesized that the best-fit models would not support a full Pangaean distribution (i.e., including all areas) for the origin of crown Squamata, but would instead show regionalization to specific areas within the fragmenting supercontinent, likely in the Northern Hemisphere where most early squamate fossils have been found. Results: Incorporating fossil data reconstructs a localized origin within Pangaea, with early regionalization of extant lineages to Eurasia and Laurasia, while Gondwanan regionalization did not occur until the middle Cretaceous for Alethinophidia, Scolecophidia, and some crown Gekkotan lineages. While the Mesozoic history of extant squamate biogeography can be summarized as a Eurasian origin with dispersal out of Laurasia into Gondwana, their Cenozoic history is complex with multiple events (including secondary and tertiary recolonizations) in several directions. As noted by previous authors, squamates have likely utilized over-land range expansion, land-bridge colonization, and trans-oceanic dispersal. Tropical Gondwana and Eurasia hold more ancient lineages than the Holarctic (Rhineuridae being a major exception), and some asymmetries in colonization (e.g., to North America from Eurasia during the Cenozoic through Beringia) deserve additional study. Future studies that incorporate fossil branches, rather than as node constraints, into the reconstruction can be used to explore this history further.

Selective extinction and habitat nestedness are the main drivers of lizard nestedness in the Zhoushan Archipelago.

Nested subset pattern (nestedness) is an important part of the theoretical framework of island biogeography and community ecology. However, most previous studies often used nestedness metrics or randomization algorithms that are vulnerable to type I error. In this study, we investigated the nestedness of lizard assemblages on 37 islands in the Zhoushan Archipelago, China. We used the line-transect method to survey species occurrence, abundance, and habitat types of lizards on 37 islands during 2 breeding seasons in 2021 and 2022. We applied the nested metric WNODF and the conservative rc null model to control for type I error and quantify the significance of nestedness. Spearman rank correlations were used to evaluate the role of 4 habitat variables (island area, 2 isolation indices, and habitat diversity) and 4 ecological traits (body size, geographic range size, clutch size, and minimum area requirement) in generating nestedness. The results of WNODF analyses showed that lizard assemblages were significantly nested. The habitat-by-site matrix estimated by the program NODF was also significantly nested, supporting the habitat nestedness hypothesis. The nestedness of lizard assemblages were significantly correlated with island area, habitat diversity, clutch size, and minimum area requirement. Overall, our results suggest that selective extinction and habitat nestedness were the main drivers of lizard nestedness in our system. In contrast, the nestedness of lizard assemblages was not due to passive sampling or selective colonization. To maximize the number of species preserved, our results indicate that we should protect both large islands with diverse habitats and species with large area requirement and clutch size.

Hidden social complexity behind vocal and acoustic communication in non-avian reptiles.

Social interactions are inevitable in the lives of most animals, since most essential behaviours require interaction with conspecifics, such as mating and competing for resources. Non-avian reptiles are typically viewed as solitary animals that predominantly use their vision and olfaction to communicate with conspecifics. Nevertheless, in recent years, evidence is mounting that some reptiles can produce sounds and have the potential for acoustic communication. Reptiles that can produce sound have an additional communicative channel (in addition to visual/olfactory channels), which could suggest they have a higher communicative complexity, the evolution of which is assumed to be driven by the need of social interactions. Thus, acoustic reptiles may provide an opportunity to unveil the true social complexity of reptiles that are usually thought of as solitary. This review aims to reveal the hidden social interactions behind the use of sounds in non-avian reptiles. Our review suggests that the potential of vocal and acoustic communication and the complexity of social interactions may be underestimated in non-avian reptiles, and that acoustic reptiles may provide a great opportunity to uncover the coevolution between sociality and communication in non-avian reptiles. This article is part of the theme issue 'The power of sound: unravelling how acoustic communication shapes group dynamics'.

Pathogenic Potential of Opportunistic Gram-Negative Bacteria Isolated from the Cloacal Microbiota of Free-Living Reptile Hosts Originating from Bulgaria.

Reptiles are known to be asymptomatic carriers of various zoonotic pathogens. A number of Gram-negative opportunistic commensals are causative agents of bacterial infections in immunocompromised or stressed hosts and are disseminated by reptiles, whose epidemiological role should not be neglected. Since most studies have focused on exotic species, in captivity or as pet animals, the role of wild populations as a potential source of pathogens still remains understudied. In the present study, we isolated a variety of Gram-negative bacteria from the cloacal microbiota of free-living lizard and tortoise hosts (Reptilia: Sauria and Testudines) from the Bulgarian herpetofauna. We evaluated their pathogenic potential according to their antibiotic susceptibility patterns, biofilm-forming capacity, and extracellular production of some enzymes considered to play roles as virulence factors. To our knowledge, the phenotypic manifestation of virulence factors/enzymatic activity and biofilm formation in wild reptile microbiota has not yet been widely investigated. All isolates were found to be capable of forming biofilms to some extent and 29.6% of them could be categorized as strong producers. Two strains proved to be excellent producers. The majority of the isolated strains showed extracellular production of at least one exoenzyme. The most pronounced pathogenicity could be attributed to the newly isolated Pseudomonas aeruginosa strain due to its multiresistance, excellent biofilm formation, and expression of exoenzymes.

Comparative anatomy and evolution of the atlantoaxial complex in the fossorial lineage Amphisbaenia (Squamata: Lacertoidea).

The atlas and axis are the first two vertebrae from the cervical series; these two vertebrae are responsible for neck flexion, extension, and rotation movements, while providing insertion points for muscles and tendons. Amphisbaenia is a group of fossorial squamates known for having four distinctive head shapes, which are related to different excavation methods. However, little is known about the relationship between these different digging patterns and the anatomy and evolution of the atlantoaxial complex. In this study, we used computed microtomography data to describe in detail of the atlantoaxial complex for 15 species, belonging to all six current families of Amphisbaenia. Furthermore, we evaluate evolutionary scenarios of selected characters related to the atlantoaxial complex in the most recent phylogeny for Amphisbaenia, using the criteria of parsimony and maximum likelihood. Our results indicate that the evolutionary pattern of the atlantoaxial complex presents a diversification in its morphology that is not always correlated with the shape of the head. This analysis reinforces the hypothesis of remarkable morphological convergences in the evolutionary history of Amphisbaenia. Additionally, some of the characters studied may represent independent evolution through convergence in some cases (e.g., horizontal axis of the neural column) and parallelism in others (e.g., present or absent from the transverse process).

Vertebrates can be more important pollinators than invertebrates on islands: the case of Malva (=Lavatera) arborea L.

Premise of the study: On islands, flowering plants tend to be more generalist in their pollination needs, as insects (the main pollinators of flowering plants) are underrepresented in these ecosystems compared to the mainland. In addition, some vertebrate species that are typically insectivorous or granivorous on the mainland are forced to broaden their diet and consume other resources such as nectar or pollen on the islands. The shrub Malva arborea, with its large and colourful flowers, attracts different groups of potential pollinators. This study aimed to compare the effectiveness of vertebrates versus insects in an insular population of M. arborea and to investigate its reproductive system. Methods: For three groups of taxa (insects, birds and lizards), we assessed the two components of pollination effectiveness: (i) the quantitative component (i.e. number of visits and number of flowers contacted) through direct observations of flowers; and (ii) the qualitative component (fruit and seed set, number and size of seeds and proportion of seedling emergence) through pollinator exclusion experiments. Key results: Vertebrates (birds and lizards) were quantitatively the most effective pollinators, followed by insects. However, when all three groups visited the flowers, fruit and seed set were higher than when any of them were excluded. We also found that M. arborea has hermaphrodite flowers and is able to reproduce by autogamy, although less efficiently than when pollinated by animals. Conclusions: Both vertebrates and insects play an important role in the reproduction of M. arborea. Although the plant does not need pollinators to produce seeds, its reproductive success increases when all pollinators are allowed to visit the flowers. Besides providing new information on M. arborea, these findings may help to better understand the role of different pollinator groups in the reproduction of other plant species, especially on islands where the co-occurrence of vertebrate and invertebrate pollination in the same plant species is usual.

Report of abnormal tail regeneration of Eremias yarkandensis (Sauria: Lacertidae) and its locomotor performance.

Caudal autotomy is a phenomenon observed in many reptile taxa, and tail loss is a pivotal functional trait for reptiles, with potentially negative implications for organism fitness due to its influence on locomotion. Some lizard species can regenerate a lost tail, which sometimes can lead to the development of more than one tail (i.e., abnormal tail regeneration) in the process. However, little is currently known about the impact of abnormal tail regeneration on locomotor performance. In this study, we document abnormal tail regeneration in Eremias yarkandensis, a reptile species native to northwestern China. Additionally, we investigated the sprint speed and endurance performance of these lizards. This study provides the first report on abnormal tail regeneration and its locomotor performance on a Chinese reptile. We suggest that the abnormal regeneration of tails may contribute to the accumulation of food reserves in the species. In light of our findings, we propose that herpetologists continue to share their sporadic observations and assess the locomotor performance of species experiencing abnormal tail regeneration, further expanding our understanding of this intriguing phenomenon.

Evolutionary dynamics and geographical dispersal of Borrelia lusitaniae.

Background: Borrelia lusitaniae is a species within the complex Borrelia burgdorferi sensu lato, associated with lizards as reservoirs and Ixodes ricinus as its main vector. Borrelia lusitaniae is predominantly distributed in Central and Southeast Europe, and in countries of the Mediterranean basin, such as Portugal, Morocco, Tunisia, and Italy where this spirochete appears to infect vector ticks more frequently than other genospecies. Evolution of this zoonotic tick-borne microparasite is shaped by different environmental factors. Comprehensive phylogenetic analysis may give insight into how B. lusitaniae spreads to new geographic locations. Aim: We applied Bayesian statistical methods to B. lusitaniae multilocus sequence typing (MLST) data to study the migration routes of B. lusitaniae and its potential for further spread. Results: The discrete phylogeographic analysis placed origins of B. lusitaniae in Southeast Europe and identified at least two introductions of B. lusitaniae from Europe to North Africa. Estimated effective reproductive potential (Re), as a key indicator for a pathogen spread, suggested potential for further spread. Conclusion: The results of this study can provide beneficial information about the potential for further spread of B. lusitaniae in Europe and North Africa and estimation of necessity for the development of strategies to monitor and control Lyme borreliosis.

Predator-driven behavioural shifts in a common lizard shape resource-flow from marine to terrestrial ecosystems.

Foraging decisions shape the structure of food webs. Therefore, a behavioural shift in a single species can potentially modify resource-flow dynamics of entire ecosystems. To examine this, we conducted a field experiment to assess foraging niche dynamics of semi-arboreal brown anole lizards in the presence/absence of predatory ground-dwelling curly-tailed lizards in a replicated set of island ecosystems. One year after experimental translocation, brown anoles exposed to these predators had drastically increased perch height and reduced consumption of marine-derived food resources. This foraging niche shift altered marine-to-terrestrial resource-flow dynamics and persisted in the diets of the first-generation offspring. Furthermore, female lizards that displayed more risk-taking behaviours consumed more marine prey on islands with predators present. Our results show how predator-driven rapid behavioural shifts can alter food-web connectivity between oceanic and terrestrial ecosystems and underscore the importance of studying behaviour-mediated niche shifts to understand ecosystem functioning in rapidly changing environments.

Climate warming has divergent physiological impacts on sympatric lizards.

Climate warming is expected to affect the vulnerability of sympatric species differentially due to their divergent traits, but the underlying physiological mechanisms of those impacts are poorly understood. We conducted field warming experiments (present climate vs. warm climate) using open-top chambers to determine the effects of climate warming on active body temperature, oxidative damage, immune competence, growth and survival in two sympatric desert-dwelling lizards, Eremias multiocellata and Eremias argus from May 2019 to September 2020. Our climate warming treatment did not affect survival of the two species, but it did increase active body temperatures and growth rate in E. multiocellata compared to E. argus. Climate warming also induced greater oxidative damage (higher malondialdehyde content and catalase activity) in E. multiocellata, but not in E. argus. Further, climate warming increased immune competence in E. multiocellata, but decreased immune competence in E. argus, with regards to white blood cell counts, bacteria killing ability and relative expression of immunoglobulin M. Our results suggest that climate warming enhances body temperature, and thereby oxidative stress, immune competence and growth in E. multiocellata, but decreases immune competence of E. argus, perhaps as a cost of thermoregulation to maintain body temperatures under climate warming. The divergent physiological effects of climate warming on sympatric species may have profound ecological consequences if it eventually leads to changes in reproductive activities, population dynamics and community structure. Our study highlights the importance of considering interspecific differences in physiological traits when we evaluate the impact of climate warming on organisms, even for those closely-related species coexisting within the same geographical area.

Ecological factors and parity mode correlate with genome size variation in squamate reptiles.

BACKGROUND: Evidence of correlation between genome size, the nuclear haploid DNA content of a cell, environmental factors and life-history traits have been reported in many animal species. Genome size, however, spans over three orders of magnitude across taxa and such a correlation does not seem to follow a universal pattern. In squamate reptiles, the second most species-rich order of vertebrates, there are currently no studies investigating drivers of genome size variability. We run a series of phylogenetic generalized least-squares models on 227 species of squamates to test for possible relationships between genome size and ecological factors including latitudinal distribution, bioclimatic variables and microhabitat use. We also tested whether genome size variation can be associated with parity mode, a highly variable life history trait in this order of reptiles. RESULTS: The best-fitting model showed that the interaction between microhabitat use and parity mode mainly accounted for genome size variation. Larger genome sizes were found in live-bearing species that live in rock/sand ecosystems and in egg-laying arboreal taxa. On the other hand, smaller genomes were found in fossorial live-bearing species. CONCLUSIONS: Environmental factors and species parity mode appear to be among the main parameters explaining genome size variation in squamates. Our results suggest that genome size may favour adaptation of some species to certain environments or could otherwise result from the interaction between environmental factors and parity mode. Integration of genome size and genome sequencing data could help understand the role of differential genome content in the evolutionary process of genome size variation in squamates.

Phylogenetic Analyses of Lizards from the Chilean Humboldt Archipelago Reveal a New Species for the Chañaral Island (Squamata: Liolaemidae).

The Humboldt Archipelago, situated on Chile's north-central coast, is renowned for its exceptional biodiversity. However, lizards of the Liolaemus genus are a particularly understudied group in this archipelago. Liolaemus genus is divided into two clades: chiliensis and nigromaculatus. Within the nigromaculatus clade the zapallarensis group is restricted to the semi-arid and arid coastal habitats of the Atacama Desert in north-central Chile. While it has been reported that lizards from the zapallarensis group inhabit various islands within the Humboldt Archipelago, there has been limited knowledge regarding their specific species identification. To identify the lizard species inhabiting these islands, we conducted phylogenetic analyses using a mitochondrial gene and examined morphological characteristics. Our findings reveal that lizards from the Damas, Choros, and Gaviota islands belong to Liolaemus silvai. In contrast, the lizards on Chañaral Island form a distinct and previously unrecognised group, clearly distinguishable from Liolaemus silvai. In conclusion, our study not only confirms the presence of L. silvai on the Damas, Choros, and Gaviota islands but also describes a new lizard species on Chañaral Island named Liolaemus carezzae sp. nov. These findings contribute valuable insights into the biodiversity of these islands and introduce a newly discovered endemic taxon to the region, enriching our understanding of Chile's unique island ecosystems.

The effects of early-life and intergenerational stress on the brain.

Stress experienced during ontogeny can have profound effects on the adult phenotype. However, stress can also be experienced intergenerationally, where an offspring's phenotype can be moulded by stress experienced by the parents. Although early-life and intergenerational stress can alter anatomy, physiology, and behaviour, nothing is known about how these stress contexts interact to affect the neural phenotype. Here, we examined how early-life and intergenerational stress affect the brain in eastern fence lizards (Sceloporus undulatus). Some lizard populations co-occur with predatory fire ants, and stress from fire ant attacks exerts intergenerational physiological and behavioural changes in lizards. However, it is unclear if intergenerational stress, or the interaction between intergenerational and early-life stress, modulates the brain. To test this, we captured gravid females from fire ant invaded and uninvaded populations, and subjected offspring to three early-life stress treatments: (1) fire ant attack, (2) corticosterone, or (3) a control. Corticosterone and fire ant attack decreased some aspects of the neural phenotype while population of origin and the interaction of early-life stress and population had no effects on the brain. These results suggest that early-life stressors may better predict adult brain variation than intergenerational stress in this species.

Malignant ovarian dysgerminoma in a 16-year-old leopard gecko (Eublepharis macularius).

The 16-year-old female leopard gecko (Eublepharis macularius) was presented with distended coelom and cachexia. Examination of the faecal sample ruled out the presence of protozoan parasites. A radiographic examination confirmed the presence of radiopaque foreign material in the intestine. The conservative treatment with tramadol, butylscopolamine, famotidine, vitamin B complex, and supportive fluid therapy with Hartmann solution and Duphalyte, was performed for 14 days. Ultrasonographic examination revealed the presence of a large mass adherent to the liver (with hypoechoic regions), a thin-walled cystic structure close to the liver, and coelomic effusion. Surgical exploration revealed a large mass on the right ovary. The unilateral (right) ovariectomy was performed. Histologic examination of the mass revealed dysgerminoma with an invasion of the ovarian bursa and blood vessels. Nine months after the surgery the patient was active and doing well. In reptiles, dysgerminoma is an uncommon type of neoplasia. To the best of our knowledge, this is the first case of dysgerminoma tumour diagnosed intravitally and treated successfully in lizards.

Metabolic regulation reduces the oxidative damage of arid lizards in response to moderate heat events.

Climate warming poses a significant threat to species worldwide, particularly those inhabiting arid and semi-arid regions where extreme temperatures are increasingly prevalent. However, empirical studies investigating how moderate heat events affect the physiological processes of arid and semi-arid animals are largely scarce. To address this knowledge gap, we used an arid and semi-arid lizard species (Phrynocephalus przewalskii) as a study system. We manipulated thermal environments to simulate moderate heat events (43.5 ± 0.3°C during the heating period) for lizards and examined physiological and biochemical traits related to survival, metabolism, locomotion, oxidative stress, and telomere length. We found that the body condition and survival of the lizards were not significantly affected by moderate heat events, despite an increase in body temperature and a decrease in locomotion at high test temperatures were detected. Mechanistically, we found that the lizards exhibited down-regulated metabolic rates and enhanced activities of antioxidative enzymes, resulting in reduced oxidative damage and stable telomere length under moderate heat events. Based on these findings, which indicated a beneficial regulation of fitness by physiological and biochemical processes, we inferred that moderate heat events did not have a detrimental effect on the toad-headed agama, P. przewalskii. Overall, our research contributes to understanding the impacts of moderate heat events on arid and semi-arid species and highlights the adaptive responses and resilience exhibited by the toad-headed agama in the face of climate warming.

Sex identification in juvenile and adult Indonesian blue-tongued skinks (Tiliqua gigas) through cystoscopy and accuracy of contrast radiography.

OBJECTIVE: To describe a nonsurgical endoscopic technique for sex identification in Indonesian blue-tongued skinks (Tiliqua gigas) and to assess accuracy of contrast radiography of the hemipenile/hemiclitoral pouches. ANIMALS: 42 clinically healthy Indonesian blue-tongued skinks between 6 months and 3 years old and weighing between 22 and 550 g. METHODS: Cystoscopy was performed under general anesthesia. Gonads were visualized through the transparency of the urinary bladder, and their gross morphology was described. Contrast was applied in the tail pouches before obtaining full-body radiographs. Two radiologists, blinded to the sex of the skink, evaluated the radiographs. RESULTS: Cystoscopy was achieved in all 42 skinks. Visualization of the gonads through the urinary bladder was possible in 41 (98%; 95% CI, 87% to 99%) of the skinks, with 18 of them identified as males and 23 identified as females. Median procedure time was 60 seconds (range, 25 to 180 seconds) and was not associated with procedure order (-0.69; 95% CI, -1.83 to 0.45) or with the weight (0.02 g; 95% CI, -0.07 to 1.0) or the identified sex (11.7; 95% CI, -15.07 to 38.45) of the skink. Radiographs had a sensitivity of 69.6% (95% CI, 47.1% to 86.8%) and a specificity of 75.0% (47.6% to 92.7%) to identify female skinks. All the skinks recovered uneventfully. CLINICAL RELEVANCE: Cystoscopic sex identification is feasible in Indonesian blue-tongued skinks of various age and size. Considering the difficulty in identifying their sex otherwise, this technique could provide a significant improvement in the veterinary care of this species. In this population, contrast radiographs showed limited accuracy for sex identification.