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Accurate delineation of biodiversity units is essential for effective conservation, particularly in species complexes with cryptic diversity and fragmented distributions. In this study, we investigated the genetic structure, phylogenetic relationships, and phylogeographic history of Iranian populations within the Vipera ursinii–renardi complex by analysing two mitochondrial genes (cyt b , ND4) and three nuclear loci (PRLR, BDNF, NT3) from 12 individuals sampled across the species’ known range. Multilocus phylogenetic analyses revealed two well-supported evolutionary lineages, including a previously unreported northwestern lineage (Lineage NW), which is sister to the Central Alborz lineage ( V. ebneri sensu stricto; Lineage CA). Genetic clustering and haplotype network analyses further supported the distinctiveness of these lineages, indicating limited or no gene flow between them. Molecular-clock dating suggests that the divergence between Lineage CA and Lineage NW occurred during the Late Pleistocene, likely driven by climatic oscillations that promoted ecological isolation. Based on reciprocal mitochondrial monophyly, nuclear differentiation, and geographic separation, we recognize Lineage CA and Lineage NW as two evolutionarily significant units in Iran. Recognizing and conserving this cryptic diversity is vital for preserving the evolutionary legacy of Iranian steppe vipers and for guiding effective conservation strategies under increasing environmental and anthropogenic pressures.

The karst viper (Vipera ursinii ssp.) favours high-mountain dry grasslands in southern and south-eastern Croatia. It is medically less important than other Vipera species, because of its remote habitat and the very small amount of venom that it injects by its relatively short fangs. The scientific literature on Vipera ursinii deals mostly with the morphology, ecology and distribution range of this snake, due to the species’ conservation issues, while the toxinological aspects of its venom have not so far been investigated. Here we report on the composition and biological activity of the Vipera ursinii ssp. venom. Using a proteomics approach, we have identified 25 proteins in the venom that belong to seven protein families: snake venom metalloproteinase, serine protease, secreted phospholipase A2, cysteine-rich secretory protein, snake C-type lectin-like protein, serine protease inhibitor and nerve growth factor. The Vipera ursinii ssp. venom was found to be distinctively insecticidal. Its lethal toxicity towards crickets was more than five times greater than that of Vipera ammodytes ammodytes venom, while the opposite held in mice. Interestingly, the mode of dying after injecting a mouse with Vipera ursinii ssp. venom may suggest the presence of a neurotoxic component. Neurotoxic effects of European vipers have so far been ascribed exclusively to ammodytoxins and ammodytoxin-like basic secreted phospholipases A2. Structural and immunological analyses of the Vipera ursinii ssp. venom, however, confirmed that ammodytoxin-like proteins are not present in this venom.

Aim: The aim of this study was to elucidate the phylogeographical pattern of taxa composing the Vipera ursinii complex, for which the taxonomic status and the dating of splitting events have been the subject of much debate. The objectives were to delimit potential refugia and to date splitting events in order to suggest a scenario that explains the diversification of this species complex. Location: Western Europe to Central Asia. Methods: Sequences of the mitochondrial cytochrome b and NADH dehydrogenase subunit 4 (ND4) genes were analysed for 125 individuals from 46 locations throughout the distribution range of the complex. The phylogeographical structure was investigated using Bayesian and maximum likelihood methods. Molecular dating was performed using three calibration points to estimate the timing of diversification. Results: Eighty-nine haplotypes were observed from the concatenation of the two genes. Phylogenetic inferences supported two main groups, referred to in this study as the 'ursinii clade' and the 'renardi clade', within which several subclades were identified. Samples from Greece (Vipera ursinii graeca) represented the first split within the V. ursinii complex. In addition, three main periods of diversification were revealed, mainly during the Pleistocene (2.4-2.0 Ma, 1.4 Ma and 1.0-0.6 Ma). Main conclusions: The present distribution of the V. ursinii complex seems to have been shaped by Quaternary climatic fluctuations, and the Balkan, Caucasus and Carpathian regions are identified in this study as probable refugia. Our results support a south-north pattern of colonization, in contrast to the northsouth colonization previously proposed for this complex. The biogeographical history of the V. ursinii complex corroborates other biogeographical studies that have revealed an east-west disjunction (situated near the Black Sea) within a species complex distributed throughout the Palaearctic region.

The genus Vipera encompasses most species of medically significant venomous snakes of Europe, with Italy harbouring four of them. Envenomation by European vipers can result in severe consequences, but underreporting and the absence of standardised clinical protocols hinder effective snakebite management. This study provides an updated, detailed set of guidelines for the management and treatment of Vipera snakebite tailored for Italian clinicians. It includes taxonomic keys for snake identification, insights into viper venom composition, and recommendations for clinical management. Emphasis is placed on quick and reliable identification of medically relevant snake species, along with appropriate first aid measures. Criteria for antivenom administration are outlined, as well as indications on managing potential side effects. While the protocol is specific to Italy, its methodology can potentially be adapted for other European countries, depending on local resources. The promotion of comprehensive data collection and collaboration among Poison Control Centres is advocated to optimise envenomation management protocols and improve the reporting of epidemiological data concerning snakebite at the country level.

Transferability of habitat suitability models (HSMs), essential to accurately predict outside calibration conditions, has been seldom investigated at intraspecific level. We targeted Vipera ursinii ursinii, a meadow viper from southeastern France and central Italy, to assess determinants of transferability among geographically disjunct populations. We fitted HSMs upon occurrences of the Italian and French populations separately, as well as on the combined occurrence dataset. Internal transferability of HSMs, on spatially independent test data drawn from the calibration region, and their external transferability on the geographically disjunct populations were evaluated according to (a) use of full or spatially rarefied presence datasets; (b) ecology‐driven or statistics‐driven filtering of predictors; (c) modeling algorithm, testing generalized additive models and gradient boosting models; and (d) multivariate environmental novelty within test data. Niche overlap between French and Italian populations was also tested. Niche overlap was low, but niche divergence between the two populations’ clusters was not corroborated. Nonetheless, wider niche breadth and heterogeneity of background environmental conditions characterizing the French populations led to low intercluster transferability. Although models fitted on the combined datasets did not attain consistently higher internal transferability than those separately fitted for the French and Italian populations, ensemble projection from the HSMs fitted on the joint occurrences produced more consistent suitability predictions across the full range of V. u. ursinii. Spatial thinning of occurrences ameliorated internal transferability but did not affect external transferability. The two approaches to predictors filtering did not differ in transferability of the respective HSMs but led to discrepant estimated environment–occurrence relationships and spatial predictions, while the two algorithms attained different relative rankings depending on the considered prediction task. Multivariate novelty of projection sites was negatively correlated to both internal transferability and external transferability. Our findings clarify issues researchers should keep in mind when using HSMs to get predictions across geographically disjunct populations. Transferability of habitat suitability models (HSMs) has been a largely debated topic in predictive ecology for the last two decades. We investigated the factors affecting HSMs https://CRAN.R-project.org/package=ncf transferability across geographically disjunct populations using as a model system Vipera ursinii ursinii, a meadow viper from southeastern France and central Italy. Spatial thinning of occurrences improves internal transferability, while incomplete coverage of the taxon realized niche within calibration data hampers external transferability in case of multivariate novelty in test data. Algorithm chosen to fit the HSMs and the approach used to filter candidate predictors should be also taken into account to comprehensively interpret model predictions.

In a rapidly changing thermal environment, reptiles are primarily dependent on in situ adaptation because of their limited ability to disperse and the restricted opportunity to shift their ranges. However, the rapid pace of climate change may surpass these adaptation capabilities or elevate energy expenditures. Therefore, understanding the variability in thermal traits at both individual and population scales is crucial, offering insights into reptiles' vulnerability to climate change. We studied the thermal ecology of the endangered Greek meadow viper ( Vipera graeca ), an endemic venomous snake of fragmented alpine-subalpine meadows above 1600 m of the Pindos mountain range in Greece and Albania, to assess its susceptibility to anticipated changes in the alpine thermal environment. We measured preferred body temperature in artificial thermal gradient, field body temperatures of 74 individuals in five populations encompassing the entire geographic range of the species, and collected data on the available of temperatures for thermoregulation. We found that the preferred body temperature ( T p ) differed only between the northernmost and the southernmost populations and increased with female body size but did not depend on sex or the gravidity status of females. T p increased with latitude but was unaffected by the phylogenetic position of the populations. We also found high accuracy of thermoregulation in V. graeca populations and variation in the thermal quality of habitats throughout the range. The overall effectiveness of thermoregulation was high, indicating that V. graeca successfully achieves its target temperatures and exploits the thermal landscape. Current climatic conditions limit the activity period by an estimated 1278 h per year, which is expected to increase considerably under future climate scenarios. Restricted time available for thermoregulation, foraging and reproduction will represent a serious threat to the fitness of individuals and the persistence of populations in addition to habitat loss due to mining, tourism or skiing and habitat degradation due to overgrazing in the shrinking mountaintop habitats of V. graeca .

Climate change threatens species adapted to cool alpine environments, particularly ectotherms like reptiles. Small-sized grassland specialist vipers inhabit such environments in Eurasia and are highly susceptible to overheating and dehydration as global temperature rises. This study modelled activity restriction times, defined as hours when environmental temperatures exceed the thermal tolerance (i.e. not available for essential activities) of the species, for 20 grassland viper taxa to assess climate change impacts. Under future conditions, hours of activity restriction are projected to increase by 21% by the SSP1-2.6 scenario, and by 52.1% by the SSP5-8.5 scenario. Elevation and latitude significantly influenced restriction time changes, with high-altitude and northern populations predicted to be most affected. The taxa Vipera graeca and Vipera ursinii moldavica are expected to experience the greatest increase in restriction times. Despite warmer conditions potentially increasing hours within preferred thermal ranges, vipers are unlikely to exploit lower-elevation habitats due to competition and ecological constraints. These findings emphasise the urgent need for conservation strategies, including habitat preservation and connectivity, to mitigate the adverse effects of climate change on grassland vipers, particularly the most vulnerable populations.

Aim The ongoing global change makes landscape planning and management of ecological corridors crucial to preserve biodiversity. We propose a workflow optimizing the use of different data sources to convert ecological niche models (ENMs) into landscape‐focused species distribution models (SDMs), using these latter to compute ecological corridors. We infer corridors connecting present occurrence localities to future climatic refugia as well as to localities where extinct populations occurred. Also, a continuous connectivity change index is proposed to assess current–future differences. Finally, we discuss possible applications of our workflow to conservation, assessing the capability of established protected areas to preserve ecological corridors. Location Europe. Methods As case study to illustrate our framework, we use a database comprising occurrence localities of Vipera ursinii, one of the most endangered European reptiles. We obtain weighted SDMs for each of the four V. ursinii subspecies by coupling climate‐based ENMs with standardized occurrence frequencies along land use and altitude gradients through weighted averaging in GIS. We calculate current and future landscape connectivity for each subspecies based on the corresponding weighted SDM. We compare predictive performance of “traditional” ENMs, including climate, land use and topography as predictors and weighted SDMs. Results Weighted SDMs outperform ENMs, according to Boyce index. SDMs are used to infer connectivity, predicted to decrease in all future scenarios for V. ursinii, and assess where connections may favour movements of individuals to, for example, future suitable areas. Generally, protected areas are predicted to cover low‐connectivity territories. Main conclusions The proposed “couple‐and‐weigh” approach could represent a helpful tool to investigate biogeography, conservation and landscape planning topics, as it permits to capitalize on occurrence records and accessible environmental predictors by narrowing the target species’ potential distribution, estimated within “traditional” ENMs, to the realized one through post‐modelling GIS analyses, which in turn improves estimation of friction maps used to infer connectivity.

Anthropogenic climate change poses a significant threat to species on the brink of extinction. Many non-avian reptiles are endangered, but uncovering their vulnerability to climate warming is challenging, because this requires analyzing the climate sensitivity of different life stages and modeling population growth rates. Such efforts are currently hampered by a lack of long-term life-history data. In this study, we used over 3 decades of mark-recapture data from a natural population of the endangered meadow viper (Vipera ursinii ursinii) to unravel the patterns of temporal variation in reproductive traits, the local climatic determinants of inter-annual variation in reproduction, and the potential buffering effects of life cycle on population growth rate. We found significant inter-annual variation in body growth, gestation length, post-parturition body condition, clutch success, and offspring traits at birth, while reproductive effort showed little temporal variation. Temperature during gestation was the most critical factor, reducing gestation length and increasing both clutch success and post-parturition body condition. In contrast, neither air humidity nor global radiation affected reproductive outcomes. This population had a negative growth rate with minimal temporal variation, indicating a rapid decline largely independent of climatic conditions. Overall, the viper’s life-history traits appeared to be buffered against temporal variation in climatic conditions, with this declining population potentially benefiting on the short term from rising local temperatures.

We present the discovery of the mummified remains of a Vipera ursinii rakosiensis collected in 1962 from a previously unknown occurrence location in Hungary. The first author took pictures of the specimen back in 2003; there is no information about the current whereabouts of the remains.