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.

Paedomorphosis and retention of juvenile diet lead speciation in a group of Neotropical snakes (Colubroides-Philodryadini).

Dipsadidae is one of the largest clades of extant reptiles, showing an impressive morphological and ecological diversity. Despite this fact, the developmental processes behind its diversity are still largely unknown. In this study, we used 3D reconstructions based on micro-CT data and geometric morphometrics to evaluate the skull morphology of Philodryas agassizii, a small, surface-dwelling dipsadid that consume spiders. Adult individuals of P. agassizii exhibit a cranial morphology frequently observed in juveniles of other surface-dwelling colubroideans, represented in our analysis by its close relative Philodryas patagoniensis. Large orbits, gibbous neurocranial roof and a relatively short jaw complex are features present in juveniles of the latter species. Furthermore, we performed an extensive survey about diet of P. patagoniensis in which we detected an ontogenetic dietary shift, indicating that arthropods are more frequently consumed by juveniles of this dietary generalist. Thus, we infer that P. agassizzii retained not only the ancestral juvenile skull morphology but also dietary preferences. This study reveals that morphological changes driven by heterochronic changes, specifically paedomorphosis, influenced the retention of ancestral life history traits in P. agassizii, and therefore promoted cladogenesis. In this way, we obtained first evidence that heterochronic processes lead speciation in the snake megadiverse clade Dipsadidae.

A new species of the genus Paradistomum (Platyhelminthes: Trematoda) from Iwasaki's snail-eating snake Pareas iwasakii, with a note on morphological variations of Paradistomum megareceptaculum (Tamura, 1941).

Dicrocoeliid trematodes were detected from Iwasaki's snail-eating snake Pareas iwasakii in Iriomote Island, Okinawa Prefecture, Japan, and described as a new species Paradistomum dextra n. sp. in the present study. This new species can be distinguished from the type series of the other members of the genus based on size of eggs and morphological characteristics of body, oral and ventral suckers, and reproductive organs. However, the new species was hard to distinguish from Paradistomum megareceptaculum infecting snakes in Japan, including Iriomote Island where is the type locality of the new species, because it is closely similar to some part of the broad range of morphological variations in P. megareceptaculum. On the other hand, a partial sequence of 28S ribosomal DNA clearly distinguished these two species. Moreover, the new species' host snake Pareas iwasakii is reported to exclusively feed on land snails while host snakes of P. megareceptaculum feed on small vertebrates, indicating that the new species is also ecologically different from P. megareceptaculum. We also redescribed P. megareceptaculum based on adults sampled in this study and past studies to record the morphological variations of this species.

Phylogenomics of Psammodynastes and Buhoma (Elapoidea: Serpentes), with the description of a new Asian snake family.

Asian mock vipers of the genus Psammodynastes and African forest snakes of the genus Buhoma are two genera belonging to the snake superfamily Elapoidea. The phylogenetic placements of Psammodynastes and Buhoma within Elapoidea has been extremely unstable which has resulted in their uncertain and debated taxonomy. We used ultraconserved elements and traditional nuclear and mitochondrial markers to infer the phylogenetic relationships of these two genera with other elapoids. Psammodynastes, for which a reference genome has been sequenced, were found, with strong branch support, to be a relatively early diverging split within Elapoidea that is sister to a clade consisting of Elapidae, Micrelapidae and Lamprophiidae. Hence, we allocate Psammodynastes to its own family, Psammodynastidae new family. However, the phylogenetic position of Buhoma could not be resolved with a high degree of confidence. Attempts to identify the possible sources of conflict in the rapid radiation of elapoid snakes suggest that both hybridisation/introgression during the rapid diversification, including possible ghost introgression, as well as incomplete lineage sorting likely have had a confounding role. The usual practice of combining mitochondrial loci with nuclear genomic data appears to mislead phylogeny reconstructions in rapid radiation scenarios, especially in the absence of genome scale data.

Largest known madtsoiid snake from warm Eocene period of India suggests intercontinental Gondwana dispersal.

Here we report the discovery of fossils representing partial vertebral column of a giant madtsoiid snake from an early Middle Eocene (Lutetian, ~ 47 Ma) lignite-bearing succession in Kutch, western India. The estimated body length of ~ 11-15 m makes this new taxon (Vasuki indicus gen et sp. nov.) the largest known madtsoiid snake, which thrived during a warm geological interval with average temperatures estimated at ~ 28 °C. Phylogenetically, Vasuki forms a distinct clade with the Indian Late Cretaceous taxon Madtsoia pisdurensis and the North African Late Eocene Gigantophis garstini. Biogeographic considerations, seen in conjunction with its inter-relationship with other Indian and North African madtsoiids, suggest that Vasuki represents a relic lineage that originated in India. Subsequent India-Asia collision at ~ 50 Ma led to intercontinental dispersal of this lineage from the subcontinent into North Africa through southern Eurasia.

Contribution of host species and pathogen clade to snake fungal disease hotspots in Europe.

Infectious diseases are influenced by interactions between host and pathogen, and the number of infected hosts is rarely homogenous across the landscape. Areas with elevated pathogen prevalence can maintain a high force of infection and may indicate areas with disease impacts on host populations. However, isolating the ecological processes that result in increases in infection prevalence and intensity remains a challenge. Here we elucidate the contribution of pathogen clade and host species in disease hotspots caused by Ophidiomyces ophidiicola, the pathogen responsible for snake fungal disease, in 21 species of snakes infected with multiple pathogen strains across 10 countries in Europe. We found isolated areas of disease hotspots in a landscape where infections were otherwise low. O. ophidiicola clade had important effects on transmission, and areas with multiple pathogen clades had higher host infection prevalence. Snake species further influenced infection, with most positive detections coming from species within the Natrix genus. Our results suggest that both host and pathogen identity are essential components contributing to increased pathogen prevalence.

Knowledge and practices related to snakebite prevention, China: a cross-sectional study.

Objective: To assess knowledge and practices related to snakebite prevention among Chinese residents. Methods: By using a multistage random sampling approach augmented by snowball sampling, we surveyed residents from 10 provinces, one municipality and one autonomous region south of the Yangtze River Basin between May 2022 and February 2023. We supplemented the data with a national online survey. We used a χ2-test to identify differences in knowledge and behaviour across various demographic characteristics. We conducted multifactor logistic regression analyses to evaluate factors potentially influencing snakebite knowledge and practices. Findings: We obtained 55 775 valid survey responses, 16 200 respondents from the face-to-face survey and 39 575 respondents from the online survey. Only 25.7% (14 325) respondents demonstrated adequate knowledge about snakebites whereas 25.6% (14 295) respondents knew basic first-aid practices or preventive behaviours. Age, marital status, educational attainment, occupation, type of residence and frequency of exposure to nature are significant independent variables affecting snakebite knowledge (P-values: < 0.05). On the other hand, gender, age, marital status, educational attainment, occupation and type of residence were significant independent variables affecting the behaviour of snakebite prevention and first aid (P-values: < 0.05). Conclusion: There is a notable shortfall in knowledge, first aid and preventive behaviours among Chinese residents regarding snakebites. Misguided first aid practices can severely compromise the effectiveness of evidence-based therapeutic interventions. Consequently, improving health education concerning snakes and snakebites in this population is needed.

Harnessing the Cross-Neutralisation Potential of Existing Antivenoms for Mitigating the Outcomes of Snakebite in Sub-Saharan Africa.

Over 32,000 individuals succumb to snake envenoming in sub-Saharan Africa (sSA) annually. This results from several factors, including a lack of antivenom products capable of neutralising the venoms of diverse snake species in this region. Most manufacturers produce polyvalent antivenoms targeting 3 to 16 clinically important snake species in sSA. However, specific products are unavailable for many others, especially those with a restricted geographic distribution. While next-generation antivenoms, comprising a cocktail of broadly neutralising antibodies, may offer an effective solution to this problem, given the need for their clinical validation, recombinant antivenoms are far from being available to snakebite victims. One of the strategies that could immediately address this issue involves harnessing the cross-neutralisation potential of existing products. Therefore, we assessed the neutralisation potency of PANAF-Premium antivenom towards the venoms of 14 medically important snakes from 13 countries across sSA for which specific antivenom products are unavailable. Preclinical assays in a murine model of snake envenoming revealed that the venoms of most snake species under investigation were effectively neutralised by this antivenom. Thus, this finding highlights the potential use of PANAF-Premium antivenom in treating bites from diverse snakes across sSA and the utility of harnessing the cross-neutralisation potential of antivenoms.

Knowledge in identifying venomous snakes and first aid methods of snakebites among nursing students: A cross-sectional study.

BACKGROUND: Snakebites are a dangerous and significant medical emergency that occurs worldwide. The World Health Organization has recommended that teaching and training in the prevention and management of snakebites be included in the curriculum of nursing schools and other educational activities. Identification of venomous snakes and first aid would be more critical in the prevention of occupational danger worldwide. This study aims to assess the knowledge in identifying venomous snakes, snakebites, and first aid methods of snakebites among nursing students in the Southern Province of Sri Lanka. METHODS: A descriptive, cross-sectional study was performed among 425 nursing students who were studying in different educational settings: undergraduates at the University of Ruhuna, and nursing students in the three schools of nursing in Galle, Matara, and Hambantota. Data were gathered by incorporating a pre-tested self-administered questionnaire after obtaining institutional permission. The total score of whole knowledge ranged from 0 to 34 for the identification of venomous snakes. Data collection was performed after obtaining ethical clearance from the Ethics Review Committee, Faculty of Allied Health Sciences, University of Ruhuna, Sri Lanka. RESULTS: Most of the students (82.6%) were in the 24-26 age category and the majority were females. Most of the sample (64.7%) had low knowledge of identifying venomous snakes. A higher percentage of students (57.4%) had a sufficient level of knowledge about first aid methods associated with snakebites and 169 participants (39.8%) had a high level of knowledge regarding first aid methods. Further, a significant impact on students' knowledge and knowledge of first aid methods was reported. CONCLUSION AND RECOMMENDATION: The overall knowledge of identifying venomous snakes among the nursing students was inadequate. However, the knowledge about the first aid methods was at a moderate level. Strategies are needed to improve knowledge in identifying venomous snakes and first aid methods of snakebites amongst nursing students in both educational settings.

Comparative electron microscopy study of spermatozoa in snakes (Lepidosauria, Squamata).

The ultrastructure of snake sperm has received substantial attention primarily because snakes exhibit considerable variability in reproductive characteristics between species, with a wide range of mating systems and reproductive behaviors. Variability of sperm morphology among snake species may be associated with the reproductive strategies of each taxon, such as competition or sperm storage. We provide a detailed description of the sperm ultrastructure of nine snake species (Anilius scytale, Tropidophis paucisquamis, Bothrops jararaca, Oxyrhopus guibei, Dipsas mikanii, Micrurus corallinus, Xenopholis scalaris, Acrochordus javanicus, and Cylindrophis ruffus) and compared this with sperm data from the literature for the following taxa: Liotyphlops beui, Amerotyphlops reticulatus, Trilepida koppesi, Anilios waitii, Anilios endoterus, Aspidites melanochephalus, Boa constrictor amarali, Corallus hortulana, Epicrates cenchria, Boa constrictor occidentalis, Eryx jayakari, Micrurus corallinus, Micrurus surinamensis, Micrurus frontalis, Micrurus altirostris, Oxyuranus microlepidotus, Bothrops alternatus, Bothrops diporus, Crotalus durissus, Agkistrodon contortrix, Vipera aspis, Boiga irregularis, Zamenis schrenckii, Zamenis scalaris, Stegonotus cuculatus, Nerodia sipedon, Liodytes pygaea, and Myrrophis chinensis. We found twelve polymorphic characters in the ultrastructure of sperm among the described snakes. Our work supports the importance of ultrastructural analysis of sperm morphology to understand snake reproduction, and provides sperm-derived morphological characters for phylogenetic analysis.

Climate change-induced shifts in survival and size of the worlds' northernmost oviparous snake: A 68-year study.

Because of their dependence on ambient temperature ectothermic animals can serve as sentinels of conservation problems related to global warming. Reptiles in temperate areas are especially well suited to study such effects, as their annual and daily activity patterns directly depend on ambient temperature. This study is based on annual data spanning 68 years from a fringe population of Grass Snakes (Natrix natrix), which is the world's northernmost oviparous (egg-laying) reptile, and known to be constrained by temperature for reproduction, morphology, and behavior. Mark-recapture analyses showed that survival probability was generally higher in males than in females, and that it increased with body length. Body condition (scaled mass index) and body length increased over time, indicative of a longer annual activity period. Monthly survival was generally higher during winter (i.e., hibernation) than over the summer season. Summer survival increased over time, whilst winter survival decreased, especially during recent decades. Winter survival was lower when annual maximum snow depth was less than 15 cm, implying a negative effect of milder winters with less insulating snow cover. Our study demonstrates long-term shifts in body length, body condition and seasonal survival associated with a warming climate. Although the seasonal changes in survival ran in opposite directions and though changes were small in absolute terms, the trends did not cancel out, but total annual survival decreased. We conclude that effects of a warming climate can be diverse and pose a threat for thermophilic species in temperate regions, and that future studies should consider survival change by season, preferably in a long-term approach.

Recombinant snake antivenoms get closer to the clinic.

Snakebite envenomings kill ~100 000 victims each year and leave many more with permanent sequelae. Antivenoms have been available for more than 125 years but are in need of innovation. A new study by Khalek et al. highlights broadly neutralizing human monoclonal antibodies (mAbs) that might be used to develop recombinant antivenoms with superior therapeutic benefits.

Isolation of the entomopathogenic fungus Beauveria bassiana from a skin lesion in a wild Nikolsky's viper (Vipera berus nikolskii).

Mycotic lesions of the skin of snakes are often associated with the fungus Ophidiomyces ophiodiicola, but other pathogens can cause similar signs. A skin sample from a wild Nikolsky's viper (Vipera berus nikolskii) with dermal lesions was collected in eastern Ukraine. A pure fungal culture was obtained and identified using nucleotide sequence analysis as the entomopathogenic species Beauveria bassiana sensu lato. Although Beauveria spp are considered to be non-pathogenic in vertebrates, sporadic infections have been reported. This report highlights the need to explore various pathogens when diagnosing the cause of snake integumentary lesions.

Comparative analysis of human, rodent and snake deltavirus replication.

The recent discovery of Hepatitis D (HDV)-like viruses across a wide range of taxa led to the establishment of the Kolmioviridae family. Recent studies suggest that kolmiovirids can be satellites of viruses other than Hepatitis B virus (HBV), challenging the strict HBV/HDV-association dogma. Studying whether kolmiovirids are able to replicate in any animal cell they enter is essential to assess their zoonotic potential. Here, we compared replication of three kolmiovirids: HDV, rodent (RDeV) and snake (SDeV) deltavirus in vitro and in vivo. We show that SDeV has the narrowest and RDeV the broadest host cell range. High resolution imaging of cells persistently replicating these viruses revealed nuclear viral hubs with a peculiar RNA-protein organization. Finally, in vivo hydrodynamic delivery of viral replicons showed that both HDV and RDeV, but not SDeV, efficiently replicate in mouse liver, forming massive nuclear viral hubs. Our comparative analysis lays the foundation for the discovery of specific host factors controlling Kolmioviridae host-shifting.

Socially-mediated activation in the snake social-decision-making network.

Brain areas important for social perception, social reward, and social behavior - collectively referred to as the social-decision-making network (SDN) - appear to be highly conserved across taxa. These brain areas facilitate a variety of social behaviors such as conspecific approach/avoidance, aggression, mating, parental care, and recognition. Although the SDN has been investigated across taxa, little is known about its functioning in reptiles. Research on the snake SDN may provide important new insights, as snakes have a keen social perceptual system and express a relatively reduced repertoire of social behaviors. Here, we present the results of an experiment in which ball pythons (Python regius) interacted with a same-sex conspecific for one hour and neural activation was investigated through Fos immunoreactivity. Compared to controls, snakes that interacted socially had higher Fos counts in brain areas implicated in social behavior across taxa, such as the medial amygdala, preoptic area, nucleus accumbens, and basolateral amygdala. Additionally, we found differential Fos immunoreactivity in the ventral amygdala, which facilitates communication between social brain areas. In many of these areas, Fos counts differed by sex, which may be due to increased competition between males. Fos counts did not differ in early sensory (i.e., vomeronasal) processing structures. As ball python social systems lack parental care, cooperation, or long-term group living, these results provide valuable insight into the basal functions of the vertebrate social decision-making network.

DeTox: a pipeline for the detection of toxins in venomous organisms.

Venomous organisms have independently evolved the ability to produce toxins 101 times during their evolutionary history, resulting in over 200 000 venomous species. Collectively, these species produce millions of toxins, making them a valuable resource for bioprospecting and understanding the evolutionary mechanisms underlying genetic diversification. RNA-seq is the preferred method for characterizing toxin repertoires, but the analysis of the resulting data remains challenging. While early approaches relied on similarity-based mapping to known toxin databases, recent studies have highlighted the importance of structural features for toxin detection. The few existing pipelines lack an integration between these complementary approaches, and tend to be difficult to run for non-experienced users. To address these issues, we developed DeTox, a comprehensive and user-friendly tool for toxin research. It combines fast execution, parallelization and customization of parameters. DeTox was tested on published transcriptomes from gastropod mollusks, cnidarians and snakes, retrieving most putative toxins from the original articles and identifying additional peptides as potential toxins to be confirmed through manual annotation and eventually proteomic analysis. By integrating a structure-based search with similarity-based approaches, DeTox allows the comprehensive characterization of toxin repertoire in poorly-known taxa. The effect of the taxonomic bias in existing databases is minimized in DeTox, as mirrored in the detection of unique and divergent toxins that would have been overlooked by similarity-based methods. DeTox streamlines toxin annotation, providing a valuable tool for efficient identification of venom components that will enhance venom research in neglected taxa.

Characterisation, prevalence and severity of skin lesions caused by ophidiomycosis in a population of wild snakes.

Ophidiomycosis is an emerging infectious disease affecting wild snakes in the Northern Hemisphere. Recently confirmed in Great Britain, the prevalence, severity and significance of ophidiomycosis has yet to be characterised in free-living snakes at a population level in Europe. Therefore, a population of barred grass snakes (Natrix helvetica) in eastern England was monitored for three seasons (May 2019 to October 2021), to investigate the prevalence (25.5%; 191/750 snakes) and severity of skin lesions and their aetiology. The most frequently observed skin lesion characteristics were changes in scale colour, crusting, and scale margin erosion. The majority of such lesions (96.9%; 185/191 snakes) was observed on the ventral surface along the length of the body. The severity of skin lesions was considered mild in more than half of the cases (53.1%; 98/191 snakes). Predominantly, skin lesions were observed in adult snakes (72.8%; 139/191 snakes). Combined histological examinations and qPCR tests of skin lesions from N. helvetica sloughs and/or carcasses confirmed a diagnosis of ophidiomycosis. Further targeted surveillance, supported by molecular and histological examinations to confirm skin lesion aetiology, is required to determine the extent to which our findings reflect the occurrence of ophidiomycosis in populations within wider landscapes.

Another new species of Dibamus Dumril & Bibron, 1839 (Squamata: Dibamidae) from Nui Chua National Park, Ninh Thuan Province, Vietnam.

A new species of dibamid lizard, Dibamus deimontis sp. nov., is described based on eight specimens collected from the highlands of Nui Chua Mountain within Nui Chua National Park, Ninh Thuan Province, southern Vietnam. Dibamus deimontis sp. nov. co-occurs with the recently described Dibamus tropcentr, but is recorded at higher elevations (670700 m a.s.l. vs. 200280 m a.s.l.), and in a different habitat and microhabitats than the previous species. Our study represents the first report on the near sympatric occurrence of two Dibamus species. The new species can be distinguished from other congeners by the following combination of characters: rostral, labial and nasal sutures incomplete; two to three postoculars; three to five scales on the posterior edge of infralabial; 2225 midbody scale rows; 193225 ventral scales; 4755 subcaudal scales; 115 presacral and 27 tail vertebrae (in a single male specimen examined); and maximum snout-vent length 136.2 mm. We suggest this species should be considered as Vulnerable (VU) following the IUCNs Red List categories. Our study brings the number of species in the genus Dibamus to 26; Dibamus deimontis sp. nov. is the eighth species of Dibamus recorded in Vietnam, and underlines the importance of the country as a local center of reptilian diversity in Southeast Asia.

On the status of Helicops wettsteini Amaral, 1929, a senior synonym of Hypsiscopus murphyi (SERPENTES: Homalopsidae).

We redescribe the name-bearing types of Helicops wettsteini Amaral, 1929, a species described from central Costa Rica. However, while Helicops wettsteini has been considered in the literature a synonym of Hypsiscopus plumbeus (previously Enhydris plumbea), we present here morphological evidence for considering Helicops wettsteini a synonym of the recently described species Hypsiscopus murphyi Bernstein, Voris, Stuart, Phimmachak, Seateun, Sivongxay, Neang, Karns, Andrews, Osterhage, Phipps & Ruane, 2022, which hence becomes a junior subjective synonym of Hypsiscopus wettsteini (Amaral, 1929).

Expanded morphological description of the Drogons False Garden Lizard, Pseudocalotes drogon (Squamata: Agamidae) from Peninsular Malaysia.

Species in the genus Pseudocalotes are generally rare and known only from a few specimens. Pseudocalotes drogon was described based on a single male from Frasers Hill, Pahang, Peninsular Malaysia. During a recent field survey at Frasers Hill, a female specimen was collected and identified as P. drogon based on morphological characters and a molecular phylogenetic analysis. The morphological description of P. drogon is expanded here, based on the male holotype and the newly collected female specimen.