Redescription of Raillietiella mottae (Raillietiellidae: Pentastomida) a generalist parasite of insectivorous lizards: phenotypic plasticity revealed by molecular data.

Pentastomida are lung parasites of a variety of hosts, mainly reptiles. Raillietiella is the genus with the largest number of species, whose taxonomic identification is still largely based on morphological data alone. In this classification, differences divide the genus into morphotypic groups. The Raillietiella parasites of insectivorous lizards, for example, are divided into "Group I" with species with hooks with sharp tips and "Group II" with posterior hooks with blunt tips. In the present work, we used molecular analysis to infer the morphological variation in hooks and size of specimens of Raillietiella mottae, parasites of several species of insectivorous lizards. Our results demonstrate that morphological differences comprise a phenotypic plasticity of a single species, whose variation is associated with host species, with females being larger and with hooks with blunt tips in nocturnal geckos and smaller ones with sharp tips in diurnal insectivorous lizards of different species, sizes, and habitats.

Epidermal Differentiation Genes of the Common Wall Lizard Encode Proteins with Extremely Biased Amino Acid Contents.

The epidermal differentiation complex (EDC) is a cluster of genes that code for protein components of cornified cells on the skin surface of amniotes. Squamates are the most species-rich clade of reptiles with skin adaptations to many different environments. As the genetic regulation of the skin epidermis and its evolution has been characterized for only a few species so far, we aimed to determine the organization of the EDC in a model species of squamates, the common wall lizard (Podarcis muralis). By comparative genomics, we identified EDC genes of the wall lizard and compared them with homologs in other amniotes. We found that the EDC of the wall lizard has undergone a major rearrangement leading to a unique order of three ancestral EDC segments. Several subfamilies of EDC genes, such as those encoding epidermal differentiation proteins containing PCCC motifs (EDPCCC) and loricrins, have expanded by gene duplications. Most of the EDPCCC proteins have cysteine contents higher than 50%, whereas glycine constitutes more than 50% of the amino acid residues of loricrin 1. The extremely biased amino acid compositions indicate unique structural properties of these EDC proteins. This study demonstrates that cornification proteins of the common wall lizard differ from homologous proteins of other reptiles, illustrating the evolutionary dynamics of diversifying evolution in squamates.

Effects of blood extraction and ecophysiological experiments on stress in adult males of Liolaemus attenboroughi.

Stress during laboratory experiments can affect the outcomes of ecophysiological studies. The serum corticosterone concentration (CORT), the leukocyte profile, heterophil/lymphocyte ratio (H/L), and the presence of blood endoparasites were analyzed as a proxy of stress and immunological state in adult males of the lizard Liolaemus attenboroughi, endemic to Patagonia, Argentina. The results of the ecophysiological variables (preferred temperature, running speed, locomotor endurance, and body condition index, BCI) were analyzed in relation to stress indicators obtained from blood samples taken at three different times: at capture, and on the third and seventh days in the laboratory. Males at capture showed a high percentage of lymphocytes and heterophils and a low of basophils, monocytes, and eosinophils. Haemogregorina-type endoparasites have been recorded in the genus Liolaemus for the first time. The proportion of infected males remained stable during captivity; however, these males showed higher CORT levels, increased percentages of basophils, and decreased percentages of lymphocytes. There was a significant increment in CORT and H/L, and a decrease in BCI during laboratory experiments, compared with baseline values at capture. The performance was not related to the CORT or the repeated blood sampling. The BCI decreased, possibly due to energy reserve mobilization caused by acute stress. This study shows that blood extraction and ecophysiological experiments over 7 days have a minor effect on the stress indicators used.

Parasites of reptiles in Iran (1922-2023): A literature review.

Reptiles are among the most diverse groups of animals, inhabiting nearly all continents and environments. Understanding their parasite biodiversity has garnered significant interest, particularly from a One Health perspective. Although the highly diverse reptile fauna of Iran, comprising 272 species i.e. 89 snakes (Serpentes), 171 lizards (Sauria), 8 turtles, 2 tortoises (Testudines), 1 crocodile (Crocodilia), and 1 worm-lizard (Amphisbaenia), there is a shortage of information about parasites. The present review is a compilation of 62 studies published from 1922 to August 2024. We present information on 56 species of reptiles from five groups (amphisbaenians, crocodiles, testudines, snakes, and lizards) and 98 parasitic taxa belonging to different protozoa and metazoa i.e. nematodes, cestodes, trematodes, acanthocephala, leeches, ticks, mites, and myiasis -producing flies. Although 63 taxa were diagnosed at the species level, 35 parasite taxa were only reported at the genus or family levels. Reviewing the literature, we found a paucity of information about endemic reptiles several of which are vulnerable species. Considering that some of the detected parasites e.g. Cryptosporidium and amoebae have serious clinical and/or public health threats molecular diagnostic techniques are needed for precise identification and understanding of the epidemiology and the potential zoonotic implications associated with parasites of reptiles. There is also a need to understand the exact distribution and host-parasite associations in different reptilian species present in Iran including the role of the reptiles as intermediate and reservoir hosts.

Microscopic structure and immunolabeling of extremely overlapped scales in some scincid, anguid, and pygopod lizards.

Skink, anguid, and pygopod lizards possess an extremely flat skin, imparting a compact and solid body and shining surface that facilitates their slider and/or fossorial movements. The present morphological study, conducted using immunohistochemistry and electron microscopy, has analyzed the microscopical morphology of extremely overlapped scales in different lizards, including species with limb reduction (scincids such as Lerista bougainvilli, Scincella lateralis, Lampropholis delicata) or legless (pygopods such as Lialis burtonis and Delma molleri and the anguid Anguis fragilis). The outer surface of the epidermis shows different micro-structures of the Oberhautchen layer containing corneous beta-proteins (CBPs) with variable immunoreactivity for these proteins. The beta-layer is relatively thick in most of these species, probably in relation to the resistance against strong mechanical forces acting on scales during the movements on harsh substrates. The scincid and anguid lizards also possess and regenerate osteoderms that reinforce scales flatness and mechanical resistance during the serpentiform or fossorial movements of these reptiles. Osteoderms are absent in pygopods. Roundish cells with a granular content are detected in the deep hinge region of scales in Lerista and Lampropholis skinks. Whether these cells may secrete substances that facilitate scale anti-friction and also determine shining of the skin surface remains to be shown.

Metazoan Endoparasites of the Gecko Phyllopezus periosus (Squamata: Phyllodactylidae) Inhabiting the Rock Cliffs of a Waterfall Canyon in the Semi-Arid Region from Northeastern Brazil.

PURPOSE: This study describes the composition and infection patterns of macro endoparasites associated with the lizard Phyllopezus periosus. METHODS: We collected 115 P. periosus over 12 expeditions on the rocky cliffs of the Missão Velha waterfall canyon in northeastern Brazil. Specimens were collected by hand, during active searches conducted between 18:00 h and 23:30 h. The infection patterns were estimated using prevalence, total abundance, and mean intensity of infection for all parasites found. RESULTS: A total of 4256 parasites were collected from 104 hosts, resulting in an exceptionally high overall prevalence of 90.4%. No significant ontogenetic difference in the prevalence of infection was detected. Although juveniles exhibited a high parasite prevalence their parasite loads were significantly lower compared to adults. Six species of endoparasites were found; among these, the nematode Spauligodon oxkutzcabiensis was the most common endoparasite in the digestive tract, while the pentastomid Raillietiella mottae was the only species found in the respiratory tract. CONCLUSIONS: Infection rates in Gekkota species appear to correlate with body size, with larger individuals generally exhibiting a higher prevalence. This pattern was also observed across separate populations of P. periosus and other geckos from distinct ecological domains. The dominance of the monoxenous S. oxkutzcabiensis highlights potential transmission mechanisms, suggesting that behaviors such as limb licking after feeding may contribute to its prevalence. The occurrence of Physalopteroides venancioi, Oswaldocruzia sp., Parapharyngodon sp., Physaloptera sp., infecting P. periosus are new records for this host. These findings significantly contribute to the knowledge of host-parasite dynamics in geckos.

Mandibular gland proteomics of the Mexican alligator lizard, Abronia graminea, and the red-lipped arboreal alligator lizard, Abronia lythrochila.

A useful approach to deepen our knowledge about the origin and evolution of venom systems in Reptilia has been exploring the vast biodiversity of this clade of vertebrates in search of orally produced proteins with toxic actions, as well as their corresponding delivery systems. The occurrence of toxins in anguimorph lizards has been demonstrated experimentally or inferred from reports of the toxic effects of the oral secretions of taxa within the Varanidae and Helodermatidae families. In the present study, we have focused on two alligator lizards of the Anguidae family, the Mexican alligator lizard, Abronia graminea, and the red-lipped arboreal alligator lizard, A. lythrochila. In addition, the fine morphology of teeth of the latter species is described. The presence of a conserved set of proteins, including B-type natriuretic peptides, cysteine-rich secretory proteins, group III phospholipase A2, and kallikrein, in submandibular gland extracts was demonstrated for both Abronia species. These proteins belong to toxin families found in oral gland secretions of venomous reptile species. This finding, along with previous demonstration of toxin-producing taxa in both paleo- and neoanguimorpha clades, provides further support for the existence of a handful of conserved toxin families in oral secretions across the 100+ million years of Anguimorpha cladogenesis.

Clinical features, surgical management and outcome of squamous and basal cell carcinoma in squamates and chelonians.

BACKGROUND: Squamous cell carcinoma (SCC) is one of the most commonly diagnosed neoplastic disorders in reptiles. Recently, however, it has been demonstrated that basal cell carcinomas (BCCs) are frequently misclassified as SCCs. Several histological SCC and BCC variants have been characterised and their classification may allow the establishment of appropriate prognosis estimation and treatment approaches. HYPOTHESIS/OBJECTIVES: To describe the clinical features and surgical outcomes of SCCs and BCCs diagnosed between 2010 and 2022 in reptiles. ANIMALS: Thirty-three captive reptiles (21 squamates and 12 chelonians). MATERIALS AND METHODS: Detailed clinical history, including staging and surgical outcomes, were performed. Statistical analysis assessed significant factors using Prism (v8.2.1). RESULTS: While SCC was predominantly diagnosed in lizards, BCC was most commonly diagnosed in chelonians, and both neoplasms mainly occurred in adult to aged, male individuals. Although the gross pathological findings were highly comparable between SCC and BCC, considerable variation could be seen according to the primary location (oral, cutaneous or epidermis of the shell). Humane euthanasia or noncurative intent surgeries were performed in a minority of the cases. Curative intent surgeries were successful in 19 of 27 cases during a 1- to 7-year follow-up period, yet recurrence was seen in 8 cases. The results of this study allowed the identification of significant high-risk prognostic factors for SCC and BCC in reptiles. CONCLUSIONS AND CLINICAL RELEVANCE: This study contributes to predicting the clinical behaviour and prognosis of distinct SCC and BCC histological variants, and selecting the most appropriate treatment protocol.

The Phylogenetic Relationships of Major Lizard Families Using Mitochondrial Genomes and Selection Pressure Analyses in Anguimorpha.

Anguimorpha, within the order Squamata, represents a group with distinct morphological and behavioral characteristics in different ecological niches among lizards. Within Anguimorpha, there is a group characterized by limb loss, occupying lower ecological niches, concentrated within the subfamily Anguinae. Lizards with limbs and those without exhibit distinct locomotor abilities when adapting to their habitats, which in turn necessitate varying degrees of energy expenditure. Mitochondria, known as the metabolic powerhouses of cells, play a crucial role in providing approximately 95% of an organism's energy. Functionally, mitogenomes (mitochondrial genomes) can serve as a valuable tool for investigating potential adaptive evolutionary selection behind limb loss in reptiles. Due to the variation of mitogenome structures among each species, as well as its simple genetic structure, maternal inheritance, and high evolutionary rate, the mitogenome is increasingly utilized to reconstruct phylogenetic relationships of squamate animals. In this study, we sequenced the mitogenomes of two species within Anguimorpha as well as the mitogenomes of two species in Gekkota and four species in Scincoidea. We compared these data with the mitogenome content and evolutionary history of related species. Within Anguimorpha, between the mitogenomes of limbless and limbed lizards, a branch-site model analysis supported the presence of 10 positively selected sites: Cytb protein (at sites 183 and 187), ND2 protein (at sites 90, 155, and 198), ND3 protein (at site 21), ND5 protein (at sites 12 and 267), and ND6 protein (at sites 72 and 119). These findings suggested that positive selection of mitogenome in limbless lizards may be associated with the energy requirements for their locomotion. Additionally, we acquired data from 205 mitogenomes from the NCBI database. Bayesian inference (BI) and Maximum Likelihood (ML) trees were constructed using the 13 mitochondrial protein-coding genes (PCGs) and two rRNAs (12S rRNA and 16S rRNA) from 213 mitogenomes. Our phylogenetic tree and the divergence time estimates for Squamata based on mitogenome data are consistent with results from previous studies. Gekkota was placed at the root of Squamata in both BI and ML trees. However, within the Toxicofera clade, due to long-branch attraction, Anguimorpha and (Pleurodonta + (Serpentes + Acrodonta)) were closely related groupings, which might indicate errors and also demonstrate that mitogenome-based phylogenetic trees may not effectively resolve long-branch attraction issues. Additionally, we reviewed the origin and diversification of Squamata throughout the Mesozoic era, suggesting that Squamata originated in the Late Triassic (206.05 Mya), with the diversification of various superfamilies occurring during the Cretaceous period. Future improvements in constructing squamate phylogenetic relationships using mitogenomes will rely on identifying snake and acrodont species with slower evolutionary rates, ensuring comprehensive taxonomic coverage of squamate diversity, and increasing the number of genes analyzed.

A rapid inventory of amphibians, squamates, and bats of Mata de Plátano Field Station and Nature Reserve, Arecibo, Puerto Rico.

Puerto Rico harbors a diverse vertebrate fauna with high levels of endemism. However, while several books for vertebrate diversity and local checklists for birds have been published, checklists of amphibians, reptiles, and bats are lacking or nonexistent at both local and regional scales. In this study, we documented the amphibian, reptile, and bat faunas at Mata de Plátano Field Station and Nature Reserve, in Arecibo, Puerto Rico. We document four species of amphibians, seven lizards, three snakes, and nine bats. Despite prior works using samples from this nature reserve, this represents the first vertebrate checklist for the Mata de Plátano Field Station and Nature Reserve. We provide additional natural history observations based on our survey results and highlight the importance of including local and regional checklists of species for downstream research and conservation.

Environmental and anthropogenic factors mediating the functional connectivity of the mesquite lizard along the eastern Trans-Mexican Volcanic Belt.

Functional connectivity, the extent to which a landscape facilitates or impedes the dispersal of individuals across the landscape, is a key factor for the survival of species. Anthropogenic activities, such as urbanization, agriculture and roads, negatively impact functional connectivity of most species, particularly low-vagility species like lizards. Here, we examine how a landscape modified by anthropogenic activities affects the functional connectivity, at both broad and fine scales, of a widely distributed generalist lizard Sceloporus grammicus in the eastern Trans-Mexican Volcanic Belt, Mexico. We estimated for the first time the species' genetic structure, gene flow and functional connectivity in agricultural and forest zones using genomic data, a comprehensive landscape characterization and novel methods including gravity models. Our results showed not only marked genetic differentiation across the study region but also that functional connectivity is maintained for tens of kilometres despite S. grammicus low vagility. Specifically, we found that substrate and air temperature facilitated connectivity over broad and fine scales, respectively, while agricultural cover, relative humidity and slope were important for connectivity and gene flow. Contrastingly, forest cover and roads favoured (broad-scale) and limited (fine-scale) connectivity, likely associated with movement facilitated by small forest patches and with thermoregulation. Altogether, these results support that S. grammicus alternates its thermoregulatory behaviour depending on the distance travelled and the habitat environmental conditions, and that it can disperse through relatively modified landscapes, mainly using agricultural zones. The information obtained is crucial to understanding the response of lizards to current anthropogenic pressures and their potential to adapt.

Staphylococci, Reptiles, Amphibians, and Humans: What Are Their Relations?

Reptiles and amphibians are largely present in many environments, including domestic areas when they are kept as pet animals. They often harbor zoonotic pathogens, which can pose a serious risk of infection for humans, mainly immunocompromised individuals, the elderly, children, and pregnant women. Several studies have been carried out to verify the role of cold-blooded animals in the epidemiology of some bacteria, mainly Salmonella, whereas scarce attention has been focused on these animals as a source of staphylococci. These bacteria are often antimicrobial-resistant and they act as opportunistic pathogens, which can cause relevant infections in humans and animals, both domestic and wild. Asymptomatic reptiles and amphibians often harbor staphylococcal strains, such as Staphylococcus aureus and coagulase-negative Staphylococcus spp.; however, these bacteria have been associated with clinical conditions that usually appear in animals under stress conditions. In all cases, greater attention should also be focused on staphylococci in cold-blooded animals due to their implications in human and veterinary medicine.

Lizard Body Temperature Acquisition and Lizard Recognition Using Artificial Intelligence.

The acquisition of the body temperature of animals kept in captivity in biology laboratories is crucial for several studies in the field of animal biology. Traditionally, the acquisition process was carried out manually, which does not guarantee much accuracy or consistency in the acquired data and was painful for the animal. The process was then switched to a semi-manual process using a thermal camera, but it still involved manually clicking on each part of the animal's body every 20 s of the video to obtain temperature values, making it a time-consuming, non-automatic, and difficult process. This project aims to automate this acquisition process through the automatic recognition of parts of a lizard's body, reading the temperature in these parts based on a video taken with two cameras simultaneously: an RGB camera and a thermal camera. The first camera detects the location of the lizard's various body parts using artificial intelligence techniques, and the second camera allows reading of the respective temperature of each part. Due to the lack of lizard datasets, either in the biology laboratory or online, a dataset had to be created from scratch, containing the identification of the lizard and six of its body parts. YOLOv5 was used to detect the lizard and its body parts in RGB images, achieving a precision of 90.00% and a recall of 98.80%. After initial calibration, the RGB and thermal camera images are properly localised, making it possible to know the lizard's position, even when the lizard is at the same temperature as its surrounding environment, through a coordinate conversion from the RGB image to the thermal image. The thermal image has a colour temperature scale with the respective maximum and minimum temperature values, which is used to read each pixel of the thermal image, thus allowing the correct temperature to be read in each part of the lizard.

hsp70 PCR-RFLP as an alternative tool to identify Sauroleishmania species.

Sauroleishmania spp. comprises one of the four Leishmania subgenera, which has been historically considered a non-pathogenic protozoan of reptiles. However, some strains appear to be transiently infective to mammals, and recent findings have detected these parasites in dogs and humans in areas where leishmaniasis is endemic. Herein, the digestion pattern of PCR-RFLP of the 234 bp-hsp70 fragment was evaluated as a simpler and cheaper tool to distinguish the Sauroleishmania species from the other Leishmania subgenera. As a result, the digestion of the 234 bp-hsp70 fragments with HaeIII produced a banding pattern specific to the four Sauroleishmania strains assessed. This technique could contribute to the identification of Leishmania parasites isolated from sandflies, reptiles, or even mammals in fieldworks as an alternative to the use of laborious and expensive methodologies.

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.

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.

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.

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.

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.