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Polyploidization is a rare yet sometimes successful way for animals to rapidly create geno- and phenotypes that may colonize new habitats and quickly adapt to environmental changes. In this study, we use water frogs of the Pelophylax esculentus complex, comprising two species (Pelophylax lessonae, genotype LL; Pelophylax ridibundus, RR) and various diploid (LR) and triploid (LLR, LRR) hybrid forms, summarized as P. esculentus, as a model for studying recent hybridization and polyploidization in the context of speciation. Specifically, we compared the geographic distribution and genetic diversity of diploid and triploid hybrids across Europe to understand their origin, maintenance and potential role in hybrid speciation. We found that different hybrid and parental genotypes are not evenly distributed across Europe. Rather, their genetic diversity is structured by latitude and longitude and the presence/absence of parental species but not of triploids. Highest genetic diversity was observed in central and eastern Europe, the lowest in the northwestern parts of Europe. This gradient can be explained by the decrease in genetic diversity during postglacial expansion from southeastern glacial refuge areas. Genealogical relationships calculated on the basis of microsatellite data clearly indicate that hybrids are of multiple origin and include a huge variety of parental genomes. Water frogs in mixed-ploidy populations without any parental species (i.e. all-hybrid populations) can be viewed as evolutionary units that may be on their way towards hybrid speciation. Maintenance of such all-hybrid populations requires a continuous exchange of genomes between diploids and triploids, but scenarios for alternative evolutionary trajectories are discussed., (© 2015 John Wiley & Sons Ltd.)

A complex wide-range study on the haemoproteid parasites of chelonians was carried out for the first time. Altogether, 811 samples from four tortoise species from an extensive area between western Morocco and eastern Afghanistan and between Romania and southern Syria were studied by a combination of microscopic and molecular-genetic methods. Altogether 160 Haemoproteus-positive samples were gathered in the area between central Anatolia and eastern Afghanistan. According to variability in the cytochrome b gene, two monophyletic evolutionary lineages were distinguished; by means of microscopic analysis it was revealed that they corresponded to two previously described species-Haemoproteus anatolicum and Haemoproteus caucasica. Their distribution areas overlap only in a narrow strip along the Zagros Mts. range in Iran. This fact suggests the involvement of two different vector species with separated distribution. Nevertheless, no vectors were confirmed. According to phylogenetic analyses, H. caucasica represented a sister group to H. anatolicum, and both of them were most closely related to H. pacayae and H. peltocephali, described from South American river turtles. Four unique haplotypes were revealed in the population of H. caucasica, compared with seven haplotypes in H. anatolicum. Furthermore, H. caucasica was detected in two tortoise species, Testudo graeca and Testudo horsfieldii, providing evidence that Haemoproteus is not strictly host-specific to the tortoise host species., (© 2015 The Author(s) Journal of Eukaryotic Microbiology © 2015 International Society of Protistologists.)

Iran is a country characterized by high biodiversity and complex biogeographic patterns. Its diverse landscape and steep climatic gradients have resulted in significant faunal diversity and high level of endemism. To better understand these patterns, we investigated the historical environmental drivers that have shaped Iran's current geological and climatological conditions, and, consequently, have shaped the current zoological distribution patterns. Furthermore, we provide an overview of the country's zoological diversity and zoogeography by reviewing published studies on its fauna. We analyzed nearly all available catalogs, updated checklists, and relevant publications, and synthesized them to present a comprehensive overview of Iran's biodiversity. Our review reports approximately 37,500 animal species for Iran. We also demonstrated that the country serves as a biogeographic transition zone among three zoogeographical realms: the Palearctic, Oriental, and Saharo-Arabian, where distinct faunal elements intersect. This biogeographic complexity has made it challenging to delineate clear zoogeographical zones, leading to varying classifications depending on the taxon. The uplift of mountain ranges, in particular, has played a crucial role in shaping faunal diversity by serving as barriers, corridors, and glacial refugia. These mountains are largely the result of orogeny and plate collisions during the Mesozoic and Cenozoic eras, coupled with the development of the Tethyan Sea and the uplift of several ranges during the Miocene. Despite these insights, our understanding of biodiversity distribution in Iran remains incomplete, even for some well-studied taxa, such as certain vertebrate families and arthropods. We highlight the existing gaps in knowledge regarding zoogeographical patterns and propose approaches to address these gaps, particularly concerning less-studied species and the highly diverse group of insects. [ABSTRACT FROM AUTHOR]

Gametogenesis produces gametes as a piece of genetic information transmitted to the offspring. While during sexual reproduction, progeny inherits a mix of genetic material from both parents, asexually reproducing organisms transfer a copy of maternal or paternal DNA to the progeny clonally. Parthenogenetic, gynogenetic and hybridogenetic animals have developed various mechanisms of gametogenesis, however, their inheritance is not fully understood. Here, we focused on the inheritance of asexual gametogenesis in hybrid Pelophylax esculentus (RL), emerging after crosses of P. lessonae (LL) and P. ridibundus (RR). To understand the mechanisms of gametogenesis in hybrids, we performed three-generation experiments of sexual P. ridibundus females and hybrids from all-male hybrid populations. Using fluorescent in situ hybridization, micronuclei analysis, flow cytometry and genotyping, we found that most adult hybrid males simultaneously produced two types of clonal sperm. Also, most male tadpole progeny in two successive backcrossed generations simultaneously eliminated L and R parental genomes, while some progeny produced only one type of sperm. We hypothesize that the reproductive variability of males producing two kinds of sperm is an adaptive mechanism to reproduce in mixed populations with P. ridibundus and may explain the extensive distribution of the all-male lineage across the European River Basin. [ABSTRACT FROM AUTHOR]

We here describe abundant new fossil material of amphibians and reptiles from different late Neogene localities of northern Greece: the Early Pliocene (MN 14) of Spilia 0, Spilia 1, and Spilia 2; the Early Pliocene (MN 15) of Spilia 3, Spilia 4, Spilia 5, and Vevi; and the Late Miocene or Pliocene of Chalicorrema and Rema Marmara. These new late Neogene herpetofaunas are highly diverse, documenting a considerably rich herpetofauna allowing the identification of at least two salamander, seven frog, two turtle, seven lizard, and eight snake taxa. Salamanders are represented by the salamandrid genera Ommatotriton and Ichthyosaura. Frogs are represented by the bombinatorid Bombina, the discoglossids Latonia cf. ragei and Latonia sp., the pelobatid Pelobates aff. praefuscus and Pelobates sp., the ranids Pelophylax and Rana cf. dalmatina, the hylid Hyla gr. arborea, and the bufonid Bufotes gr. viridis. Turtles are represented by the emydid Emys and an indeterminate geoemydid. Lizards are represented by the scincid Ophiomorus, two lacertids (one of which potentially pertaining to Lacerta), amphisbaenians, agamids, the anguid Pseudopus, and a potential varanid. Snakes are represented by the erycid Eryx, the natricid Natrix aff. rudabanyaensis, a small-sized elapid, an "Oriental viper", the colubriforms Periergophis and Paraxenophis, as well as two further distinct but still indeterminate morphotypes of colubriforms. For the material from Spilia tentatively referred to Ommatotriton, this is only the third occurrence in the fossil record globally. The new material of Ichthyosaura and Bombina mark the first documentation of these genera in the Greek fossil record. Abundant cranial and postcranial material from Spilia is tentatively referred to Latonia ragei, a taxon previously known from the Early Miocene of Western Europe. The new record of Pelobates represents the oldest documented occurrences of the genus in the Greek fossil record. Interestingly, the Pelobates from Spilia bears much resemblance to an extinct taxon, Pelobates aff. praefuscus, which is otherwise known from the Late Miocene of the Caucasus, and not to the extant species that currently inhabits the area. The identification of Pelophylax and Rana adds to the rather poor Neogene record of ranids from Greece. Particularly for the case of Rana cf. dalmatina from Spilia 4, this corresponds to the only documented occurrence of this extant taxon in the Greek fossil record. Similarly, the identification of Hyla gr. arborea in Spilia 1, Spilia 3, and Spilia 4, marks only the third documented occurrence of this genus in the Greek fossil record. The Bufotes material from Spilia 1, Spilia 3, and Spilia 4 represents the first documented fossil occurrence of the extant Bufotes viridis complex in Greece. The material of Emys gr. orbicularis from Vevi marks the only known pre-Quaternary record of the genus in Greece and one of the only few Neogene members of the genus known from Europe. The scincid Ophiomorus is identified in Spilia 4, known from several cranial and postcranial remains, well outside the extant range of the genus. Practically, the Spilia Ophiomorus is only the fourth known fossil occurrence of this extant genus globally and also represents one of its oldest known occurrences. Material from Spilia 1 and Spilia 3 is tentatively referred to cf. Lacerta sp., and this would mark the first known occurrence of this emblematic extant genus in the Greek fossil record, denoting the presence of the genus since at least the Early Pliocene. The new amphisbaenian specimens from Spilia 4 add to the recently described single vertebra from the same locality, and represent the youngest occurrence of amphisbaenians from continental Eastern Europe. The new agamid material from Vevi and Chalicorrema add substantially to the record of this group, which was in Greece so far known exclusively from the latest Miocene/earliest Pliocene of Maramena. A similarly important addition is the record of Pseudopus from Spilia 4, as this genus had been known in the Greek fossil record only from a very few localities. The potential varanid from Spilia represents one of the few Pliocene occurrences of this group in Europe. We identify Natrix aff. rudabanyaensis among the material from Spilia 0, Spilia 4, and Vevi, adding to the previously known record of this taxon from Maramena, however, its precise species level assignment should await a revision of Neogene European Natrix spp. The find of a small elapid from Spilia 4 represents the youngest occurrence of a coral snake from Europe, extending their statigraphic range up to the Early Pliocene (MN 14). Periergophis and Paraxenophis, two bizarre snakes, so far exclusively known from their type locality in Maramena, could be also present here, even if tentatively identified. Most notably, the new material from these localities comprises forms that are now extinct (e.g., Periergophis and Paraxenophis) or extirpated from Europe (e.g., Latonia, Varanidae, Elapidae) but at the same time also loudly attests the emergence of the extant genera that dominate the extant herpetofaunas of Greece (Ichthyosaura, Bombina, Pelobates, Pelophylax, Rana, Hyla, Bufotes, Emys, Ophiomorus, Lacerta, Pseudopus, Eryx, and Natrix), for some of which their fossil record is documented here for the first time in the area. [ABSTRACT FROM AUTHOR]

Animal morphology is influenced by several factors, including gonadal development and gametogenesis. Although their effects are well documented in male/female differentiation, much less is known about same-sex effects, such as those caused by their mode of reproduction. Here, using geometric morphometric analyses, we compare two groups of all-female triploid hybrid fish Chrosomus eos × eos-neogaeus, that differ only by their sexual and asexual reproductive strategies. We demonstrate that morphological differences arise from factors inherently associated with their mode of reproduction, with results replicated in two distinct lineages and in natural and common garden environments. Such differences provide additional insight about the costs and benefits of both reproductive strategies, which have mostly been of a demographic, population genetic, or genetic nature. In particular, these findings have important implications for the ecology of asexual organisms and contribute to the study of sex evolution by adding complexity to the paradox of sex theory. Morphology associated with reproductive strategy in animals can be difficult to isolate. Here, the authors use geometric morphometrics to study triploid hybrid Chrosomus fish that reproduce sexually and asexually, identifying morphological differences associated with reproductive strategy. [ABSTRACT FROM AUTHOR]

The Spur-thighed tortoise has a large geographic distribution and can be infested by several tick species, mainly Hyalomma aegyptium. This tick has recently been confirmed as a natural carrier of numerous pathogenic, including zoonotic agents. The purpose of this study was to estimate the level of hard tick infestation in Mediterranean tortoises living in some regions of the west of Algeria (Oran, Mostaganem, and Tiaret) during the spring and summer of the year 2019. Among 39 turtles collected from the different study regions, 22 tested positive, which represents an infestation rate of 56.41 %. A total of 224 adult ticks and one nymph were collected, which can display a tick infestation intensity of 10.23 %. All of these ticks belong to a single species, which is Hyalomma aegyptium. In terms of abundance, this study showed 5.78 ticks/tortoise. Regarding sex, we have collected 198 male and 26 female ticks with a sex ratio of 7.62. The preferred attachment sites of Hyalomma aegyptium ticks were the inguinal, the limbs, and to a lesser degree, the neck. A few rare cases have been collected from the carapace. Further studies are needed to assess the impact of the intensity of H. aegyptium infestation on the tortoise's health status and to assess the potential transmission of zoonotic pathogens. [ABSTRACT FROM AUTHOR]

Despite amphibians having the least number of families in comparison with other vertebrate classes, the digenean parasites within these host species are largely predominant. To date, no investigation has been conducted on the digenean fauna of frogs in western Iran. The helminths occurrence and morphological description of this study were analyzed for the first time. A total of 54 Levant water frogs, Pelophylax bedriagae, (n = 53), and a long-legged wood frog, Rana macronemis, (n = 1) were examined. Frogs harbor in total four digenean parasites. Three taxa were characterized based on morphological features as Gorgodera cf. asiatica, G. varsoniensis, and Pleurogenoides sp. within P. bedriagae, while Haematoloechus sp. was only found in R. macronemis. Pleurogenoides sp. exhibited a moderate prevalence (23%) and intensity (7.3 ± 6.3). Gorgodera cf. asictica can be distinguished from congeners based on the body and sucker size, the ovary and testes shape, and in G. varsoviensis by having lobulated caeca and transversely elongated testes. In addition, Pleurogenoides sp. mainly differs from all its congeners by the form of its body, genital apparatus shape and Y-shaped excretory vesicles. With regard to helminths in amphibians, the existing body of literature is extensive and complex. To accurately determine the species boundaries, both morphological and molecular data are needed. We report two new records of these parasites from the region. [ABSTRACT FROM AUTHOR]

Background: Hepatozoon fitzsimonsi (Dias, 1953) is a frequently found haemogregarine of southern African tortoises. At the time of this species' reassignment from the genus Haemogregarina to Hepatozoon, developmental stages such as sporocysts and sporozoites were observed in ticks associated with H. fitzsimonsi parasitised and non-parasitised tortoises. It was thus suggested that ticks may act as the potential vectors for this parasite. However, this earlier research was unable to confirm the identity of these sporogonic stages using molecular markers. In a separate study aimed at identifying tick species parasitising South African reptiles and molecularly screening these for the presence of Hepatozoon, that study identified H. fitzsimonsi in tortoise-associated ticks. Thus, the present study aimed to revisit the potential of ticks to act as vectors for H. fitzsimonsi in tortoises using a combined microscopy and molecular approach. Methods: Specimens of Kinixys natalensis, Kinixys spekii, Kinixys zombensis and Stigmochelys pardalis were collected from Bonamanzi and Ndumo Game Reserve, South Africa. Upon capture, animals were examined for ticks, and these were collected along with blood and other tissues. Adult ticks were dissected and visceral impression slides were prepared along with thin blood and tissue smears on clean microscope slides. Smears and impression slides were stained with Giemsa, screened and micrographs of parasites were captured. Two primer sets were employed to target fragments of the 18S rRNA gene of parasites found in both tortoises and ticks and the resulting sequences were then compared with other known H. fitzsimonsi and haemogregarine sequences from the GenBank database. Results: Peripheral blood gamont and liver merogonic stages were observed in S. pardalis, while the sporogonic stages were observed in the haemocoel of Amblyomma ticks. Gamont and sporocyst stages compared morphologically with previous descriptions of H. fitzsimonsi, identifying them as this species. Phylogenetic analysis revealed that the blood and tick sequences obtained in this study clustered in a monophyletic clade comprising known H. fitzsimonsi. Conclusions: The present study provides further support for ticks acting as the vectors of H. fitzsimonsi by molecularly identifying and linking observed developmental stages in tortoises (S. pardalis) with those in the invertebrate host (Amblyomma spp.). [ABSTRACT FROM AUTHOR]

Ophidiomyces ophidiicola, the causative agent of ophidiomycosis, poses a potential threat to wild snakes worldwide. This study aimed to retrospectively investigate the prevalence of O. ophidiicola in archived snake moults collected from the San River Valley in the Bieszczady Mountains, Poland, from 2010 to 2012. Using qPCR for O. ophidiicola detection and conventional PCR for clade characterisation, we analysed 58 moults and one road-killed specimen of Zamenis longissimus and Natrix natrix. A novel combination of primers (ITS2L) was used to simultaneously confirm SYBR Green-based qPCR results and perform genotyping. O. ophidiicola has been detected from two Z. longissimus and one N. natrix specimens. The identified clade (I-B) is consistent with those found in wild snakes of eastern Europe and San River Valley, indicating that O. ophidiicola has been present in this region for at least a decade. This study underscores the value of historical samples in understanding the long-term presence of pathogens and highlights the potential role of environmental reservoirs in the persistence of O. ophidiicola. Our findings are crucial for informing conservation strategies for the endangered Aesculapian snake populations in Poland, emphasising the need for ongoing monitoring and habitat management to mitigate the potential impact of ophidiomycosis. [ABSTRACT FROM AUTHOR]

Population systems in which all three taxa of the Pelophylax esculentus complex coexist are rare and in Serbia can only be found along the Danube. Although several studies describe the diet in populations with only one taxon, there is a lack of data from mixed populations. We studied the diet in mixed populations of edible frogs at three sites for three years. We collected gut contents from 221 adult frogs using the stomach flushing method and identified 1,477 prey items. The diet consisted mainly of insects (88%). For all three taxa, the most frequently consumed prey groups were Hymenoptera (28%), Coleoptera (18%), Lepidoptera (17%) and Diptera (11%). Larger prey (in terms of length and volume) was mainly consumed by P. ridibundus, followed by P. esculentus. The smallest prey were mainly consumed by P. lessonae. However, P. ridibundus consumed fewer prey items than P. lessonae. No significant difference was found between the taxa in the staple diet, while there were differences in the less abundant prey categories, especially between P. ridibundus and P. lessonae. The narrowest trophic niche width was observed in P. lessonae, followed by P. esculentus, and the widest in P. ridibundus. Our results suggest that the diet of the three taxa is diverse and consists of a large number of invertebrate groups. However, the diet was locality-specific, with the dominance of different prey groups in different localities depending on habitat characteristics. These results confirm our expectation that the frogs of the Pelophylax esculentus complex follow an opportunistic foraging strategy, as predicted by optimal foraging theory. [ABSTRACT FROM AUTHOR]

Two water frog species: the pool frog Pelophylax lessonae (L) and the marsh frog P. ridibundus (R) occur sympatrically in Central Europe and form mixed populations (genetic systems) with their hybrid, the edible frog P. esculentus (E). The aim of the study was to assess the species composition of water frogs in urban and rural populations and compare our current findings with the results of previous studies. We surveyed the same sites that were investigated by Berger et al. in 1962–1970 (Poznań, urban landscape) and 1977–1997 (Dezydery Chłapowski Landscape Park, rural landscape). Because some ponds surveyed in the past were destroyed or dried-up, we also explored others located in the adjacent areas. We captured frogs during breeding seasons 2020 and 2021 and identified them by the nuclear marker gene SAI-1. We found three types of populations in the urban area: R-E, E-E and R-E-L and four in the rural area: R-E, L-E, E-E and R-E-L. Compared to the historical data, we found a drastic decrease in the frequency of P. lessonae in urban and rural landscapes: from 89.1% and 68% to 2.7% and 1.8%, respectively. At the same time, the frequency of P. ridibundus increased from 2.2% and 0.1% to 40% and 29.6%, respectively. A similar pattern was found for P. esculentus whose frequency increased from 8.7% and 31.9% to 57.3% and 68.6%, respectively. Additionally, we confirmed the presence of a cryptogenic Balkan water frog, P. kurtmuelleri, which was recently discovered in south-western Poland. The frequency of SAI-1 allele specific for this taxon reached 7.3%. The patterns found in both types of landscapes are in line with the current situation of both parental species in Europe. Such dynamic changes show the need for long-term monitoring of the population compositions of water frogs, what is crucial for their conservation management. [ABSTRACT FROM AUTHOR]

Ticks carry a diverse community of microorganisms including non-pathogenic symbionts, commensals, and pathogens, such as viruses, bacteria, protozoans, and fungi. The assessment of tick-borne microorganisms (TBM) in tortoises and their ticks is essential to understand their eco-epidemiology, and to map and monitor potential pathogens to humans and other animals. The aim of this study was to characterize the diversity of microorganisms found in ticks collected from the spur-thighed tortoise (Testudo graeca) in North Africa and Anatolia. Ticks feeding on wild T. graeca were collected, and pathogens were screened by polymerase chain reaction using group-specific primers. In total, 131 adult Hyalomma aegyptium ticks were collected from 92 T. graeca in Morocco (n = 48), Tunisia (n = 2), Algeria (n = 70), and Turkey (n = 11). Bacteria and protozoa detected included Hemolivia mauritanica (22.9%), Midichloria mitochondrii (11.4%), relapsing-fever borreliae (8.4%), Ehrlichia spp. (7.6%), Rickettsia spp. (3.4%), Borrelia burgdorferi s.l. (0.9%), Francisella spp. (0.9%), and Wolbachia spp. (0.8%). The characterization of Rickettsia included R. sibirica mongolitimonae (Algeria), R. aeschlimannii (Turkey), and R.africae (Morocco). Hemolivia mauritanica and Ehrlichia spp. prevalence varied significantly with the sampling region/country. We did not detect significant associations in microorganism presence within ticks, nor between microorganism presence and tick mitochondrial DNA haplogroups. This is the first report of Francisella persica-like, relapsing fever borreliae, M. mitochondrii, and Wolbachia spp. in H. aegyptium ticks collected from wild hosts from the South and Eastern Mediterranean region, and of R. sibirica mongolitimonae and R. africae in H. aegyptium from Algeria and Morocco, respectively. Given that T. graeca is a common species in commercial and non-commercial pet trade, the evaluation of the role of this species and its ticks as hosts for TBM is particularly relevant for public health., (© 2021 Wiley-VCH GmbH.)

The aim of this study is to elaborate on the prior information about Hipparchia hermione ecology, examine its behaviour and find the habitat preferences of the species in specific study areas. The obtained results contribute to better management and conservation of the species and its habitats in Slovakia. Six types of behaviour were observed, with the exception of courtship dances, mating and perching. The most common types of behaviour observed were flight and food intake. The RDA analysis of behaviour and environmental variables resulted in a positive effect of free substrate and tall-herb vegetation on H. hermione abundance. The free substrate had a significant positive effect on all types of H. hermione behaviour. The results of the RDA analysis indicate butterflies' shrub preference. The notion of shrubs' presence having a positive effect on the representation of H. hermione is supported by another data set, according to which shrub vegetation provided ideal conditions for all types of observed behaviour. The abundance and diversity of nectaring plants did not have a statistically significant effect on species representation. Due to the environmental variables, the tree crowns' significance plays a vital role. The average values are of 50%. The number of H. hermione individuals in the study areas increased with the growing number of tree trunks over 20 cm and heights over 3 m and the growing number of caterpillar host plants (Festuca ovina). The data show that Hipparchia hermione butterflies depend more on the spatial and structural arrangement of the forest than on abundance or diversity of the flora in the study areas. [ABSTRACT FROM AUTHOR]

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Studying the distribution of morphologically cryptic animal species is always a very difficult task. Because most marsh frog species (the Pelophylax ridibundus complex) are cryptic, we used molecular markers to identify them. Three marsh frog species (P. ridibundus, P. kurtmuelleri and P. cf. bedriagae) inhabit the northern part of Western Palearctic. We created a database of localities and built models of their modern distribution. These models showed that the most suitable habitats are on the north of the Mediterranean region for P. cf. bedriagae, temperate Europe for P. ridibundus, and the Balkan coastal areas for P. kurtmuelleri. The projection of the modern ecological niches under the late-Quaternary climatic conditions showed that the range of P. kurtmuelleri remained largely unchanged during the period, whereas the ranges of P. cf. bedriagae and especially P. ridibundus changed greatly over time. During the Last Glacial Maximum, the presumed range of P. cf. bedriagae covered a relatively large area in the north of the Mediterranean region and the south of European Russia. Glacial refugia of P. ridibundus were apparently located in the northern Balkans, the northern coast of the Black and Azov seas, and possibly in Western Europe. The northward long-distance post-glacial dispersal of P. ridibundus occurred from refugia in the northeastern Balkans and the Black-Azov seas region. Since the Late Pleistocene, suitable habitats for P. cf. bedriagae in southern Russia began to decline, but local habitats for P. ridibundus become more suitable. Therefore, a mosaic of populations consisting of these both species and their hybrids has now been found here. [ABSTRACT FROM AUTHOR]

Objective: The literature does not clearly specify the locations inhabited by the spurthighed tortoise (Testudo graeca) from the eastern Black Sea coast of Türkiye. This study thus aimes to reveal new locality records for T. graeca in Trabzon province. Materials and Methods: Two adult male specimens were caught from the Darıca and Konaklar neighborhoods in the respective Akçaabat and Ortahisar districts. Some of the morphological characteristics of these specimens have been recorded using a digital caliper. After taking morphometric measurements, the tortoises were returned to their habitat. No anesthetic procedure was performed on the turtles. Results: Both the Darıca and Konaklar specimens have five vertebral scutes, 11 pairs of marginal scutes, and four pairs of costal scutes on their carapace. In addition, both specimens were seen to have one undivided supracaudal scute and one nuchal scute on their carapace. The Darıca specimen has a straight carapace length (SCL) of 208.19 mm, and plastron length (PL) of 188.08 mm while the Konaklar specimen has an SCL of 216.33 mm and a PL of 196.28 mm. Conclusion: The study compared its specimens' the pholidosis and morphometric characteristics and color-pattern features with those of specimens reported in the literature. The morphological features of the Darıca and Konaklar specimens are similar to those for the samples of Testudo graeca ibera in the literature. The study's findings concluded that the samples of Darıca and Konaklar belong to the T. g. ibera subspecies. [ABSTRACT FROM AUTHOR]

Genetic diversity is a key part of biodiversity, threatened by human activities that lead to loss of gene flow and reduction of effective population sizes. Gene flow is a result of both landscape connectivity and demographic processes determining the number of dispersing individuals in space and time. Thus, the effect of human impact on processes determining the level of genetic diversity must be interpreted in the context of basic ecological conditions affecting survival and recruitment. When the intensity of human impact and habitat suitability correlate, the effect on genetic diversity and gene flow may be challenging to predict. We compared genetic diversity, gene flow and landscape resistance in two contrasting landscapes in Norway for the pond‐breeding amphibian Triturus cristatus: a highly human‐impacted, agricultural landscape with ecologically productive habitats, and a forested landscape with less productive habitats and lower levels of human impact. Our results show that genetic diversity was higher and gene flow lower within the forested landscape. Microclimatic moisture conditions and vegetation cover were important determinants of landscape resistance to gene flow within both landscapes. There were indications that landscape resistance was increased by minor roads in the forested landscape, which was not the case for the agricultural landscape, suggesting a higher vulnerability to human interference within the landscape matrix for the populations in less productive habitats. Our findings suggest that the effect of human impact on genetic diversity may not be straightforward but modulated by the ecological conditions underlying local demographic processes. Populations within both landscapes seem to be vulnerable to loss of genetic diversity, but due to different mechanisms. This has implications for the choice of relevant management actions, that is, increasing population stability may be more relevant within an agricultural landscape still permeable for dispersal, while conserving dispersal corridors may be more appropriate in the forested landscape, to avoid isolation and increased genetic drift. [ABSTRACT FROM AUTHOR]

Background Only a few comprehensive studies have been carried out on parasites in amphibians and reptiles in Ukraine. This has resulted in identifying over 100 helminth species across these vertebrate groups. However, most of the studies were performed in the 20 century and the taxonomy of many parasites and their hosts has changed ever since, in addition to the discovery of new species and registrations of species that had not been previously known for Ukraine. In recent decades, there have been very few publications on helminths from amphibian or reptile hosts in this region. Notably, just one of these recent studies is a faunistic study, providing a list of helminths found in two species of green frogs - Pelophylax ridibundus (Pallas, 1771) and Pelophylax esculentus (Linnaeus, 1758). Therefore, it is clear that publishing datasets of modern records of helminths in these vertebrate groups, based on modern taxonomy, is an essential step in further studies of their parasitic diversity. Additionally, such study is important in terms of global climate change, the growing number of possibilities of invasion of alien species (both hosts and parasites) that might potentially become a threat to native biota and growing anthropogenic pressure on local populations of hosts that affect the parasites as well. In future, this study is planned to be used for the creation of a checklist of helminths of the herpetofauna of Ukraine. The present dataset is an inventory of various species of helminths parasitising common species of the herpetofauna in central, northern, western and southern Ukraine recorded during field studies in the 2021-2023 period. New information The dataset is the first one to represent the up-to-date and unified data on helminths of reptiles and amphibians of Ukraine. Previously, records of this group of organisms with reference to their hosts were presented as several separate records within the country. Currently, this is the largest dataset presenting geocoded records of non-human-related helminths in the fauna of Ukraine. It reports helminth species from 15 hosts (205 individuals), including eight amphibians and seven reptilian species found in various Ukrainian regions. A total of 47 helminth species have been documented in the research and during 2021-2023 period on the territory of northern (Kyiv and Zhytomyr), western (Lviv, Zakarpattia Ivano-Frankivsk), central (Vinnytsia, Dnipropetrovsk, Cherkasy, Zaporizhzhia and Poltava) and southern (Odesa) regions of Ukraine. The identified helminth species belong to the following phyla: Acanthocephala (Centrorhynchidae (2), Echinorhynchidae (2)); Nematoda (Acuariidae, Anisakidae, Cosmocercidae (3), Dioctophymatidae, Gnathostomatidae (1), Kathlanidae (1), Molineidae (7), Onchocercidae (1), Pharyngodonidae (1), Rhabdiasidae (6), Strongyloididae); Platyhelminthes (Diplodiscidae (1), Diplostomidae (2), Encyclometridae (1), Haematoloechidae (1), Leptophallidae (2), Macroderidae (1), Mesocestoididae, Opisthorchiidae (2), Plagiorchiidae (3), Pleurogenidae (2), Polystomatidae (3), Proteocephalidae (1), Strigeidae (1) and Telorchiidae (3)). Only some helminths in the dataset were not identified to species level. Material is stored in the collection of the department of Parasitology of the I. I. Schmalhausen Institute of Zoology NAS of Ukraine. [ABSTRACT FROM AUTHOR]

Herein, we revise the material of the extinct taxon Pseudopus pannonicus from Central Europe, the largest known anguid lizard and iconic member of herpetofaunas from the Upper Cenozoic of Europe. The geographical position of Polgárdi 2, the type locality of P. pannonicus, as well as several other closely located important localities make Central Europe a valuable area of high interest for studies regarding European Cenozoic palaeoherpetological assemblages. We clarified the nature of the type material of Pseudopus pannonicus, showing that it consisted not only of the five specimens originally figured. Instead, the syntype series also includes a considerable number of specimens from Polgárdi 2 that were only described or figured 12 years after the original description of P. pannonicus. Detailed osteological descriptions are provided for specimens from the type series and Polish specimens, with the aid of high-resolution imaging (SEM and μCT scanning), and intraspecific variability is discussed. The articular surface with the lappet of the parietal overlapping the frontal is discussed as a character potentially relevant for the diagnosis of P. pannonicus. We updated the identification of several specimens of P. pannonicus and discussed the biogeographic implications of such revisions. In Poland, P. pannonicus is an abundant component of Neogene and early Quaternary herpetofaunas, known with certainty from the Middle Miocene of Przeworno (the oldest known occurrence of the species globally), the Early-Late Pliocene of Węże I, the Late Pliocene of Rębielice Królewskie I, the Late Pliocene of Węże II, and the Early Pleistocene of Kadzielnia (one of the youngest occurrences documented globally). An indeterminate anguine with Anguis affinities is newly reported from Rębielice Królewskie II. The taxonomic status of other large anguids from the Neogene of Europe is discussed and we conclude that most are junior synonyms of P. pannonicus. We also show that another purported synonym, that is, Ophisaurus intermedius from the Early Pleistocene of Romania, is instead a nomen nudum. We conducted phylogenetic analyses (18 taxa, 65 characters) to understand the relationship of P. pannonicus relative to other anguid representatives and anguid-related group (i.e., glyptosaurids). A single most parsimonious tree (length: 134 steps) was recovered. The clade Pseudopus is stable, comprising the two distinct sister subclades [Pseudopus laurillardi + Pseudopus ahnikoviensis] and [Pseudopus pannonicus + Pseudopus apodus]. These phylogenetic results are in accordance with previously published works., (© 2024 American Association for Anatomy.)

Freshwater vertebrate predators can exert trophic control over aquatic and littoral communities. Among these predators, post‐metamorphic anurans exhibit a biphasic trophic spectrum by foraging in both terrestrial and aquatic habitats. Many studies have described their diet through the classical taxonomic classification of prey. However, these singular diet habits imply a complex, time‐dependent, realised trophic niche in which predation pressure occurs over many consumers that fill diverse functional roles throughout the aquatic and terrestrial interface of ponds. Among anurans, marsh frogs (Pelophylax ridibundus) have been introduced outside their range in many countries and are now invading nationwide areas, particularly in western Europe. Focusing on their foraging specificities will therefore further the understanding of the trophic role of these alien taxa in pond environments that are highly colonised.We collected stomach contents from 761 marsh frogs from introduced populations in 19 ponds in southern France once a month over 4 months of their active period in the spring. The populations of marsh frogs were studied in a geographic area that was devoid of native water frogs and their origin tracks back as far as south‐eastern Europe (i.e., more than 1,000 km from the studied sites, as evidenced previously by genetic analyses).Marsh frogs exhibited generalist and opportunistic feeding strategies. The trophic niche was strongly asymmetrical and broader in the terrestrial environment than in the aquatic environment. However, predation occurred in communities of large freshwater macroinvertebrates and amphibians. Whereas the composition of the terrestrial diet exhibited strong seasonal variations, predation pressure was continuously exerted on the same aquatic organisms over time. Primary consumers and consumers at higher trophic levels frequenting aquatic benthic, vegetated, pelagic, and surface microhabitats were preyed upon, underlying the multidimensional extent of the predation spectrum.The diversified feeding strategies of alien marsh frogs highlight the extent of potential ecological control by predation on pond communities. Because of their wide trophic niche, they exert predation pressure on most pond organisms, triggering possible top‐down control of the overall aquatic communities.Our results show that the integration of the functional traits and microhabitats of consumed prey may aid in a better understanding of how predation by anurans may target specific components of pond communities. More particularly, this study raises concerns about the predatory role of introduced anurans in the context of biological invasions. [ABSTRACT FROM AUTHOR]

The southern crested newt Triturus karelinii Strauch, 1870, is widely distributed on the northern Iran plateau, where it is one of the most common newts. Mitochondrial DNA sequences (the genes cyt b and ND4) were used to determine the genetic diversity and phylogenetic position of Iranian Triturus. The results corroborate the monophyly of T. karelinii and confirm that all Iranian and Azerbaijan populations belong to the same species. Genetic distances (K2-p) between Iranian T. karelinii and other Triturus species was relatively high, (7–25% for ND4 and 8–19% for cytb). However, infraspecific genetic diversity among the Iranian populations was very low, which may indicate strong gene flow between populations. [ABSTRACT FROM AUTHOR]

Ectoparasites, such as ticks, modulate host population dynamics by impacting demographic traits. They transmit infectious agents among their hosts, posing a critical threat to animal and public health. This study aimed to characterize and analyze the Hyalomma aegyptium infestation on one of its main hosts, the spur-thighed tortoise, its effects on demographic traits, and to determine the diversity of infectious agents present in both ticks and tortoises in the Maamora forest (northwestern Morocco). Our results show that 100% of the tortoises were parasitized by adult ticks in spring, an infestation intensity of 4 ticks/tortoise (5.1 and 3.6 ticks/tortoise in males and females, respectively; 4.2 and 3.3 ticks/tortoise in gravid and non-gravid females, respectively) and an abundance ranging from 1 to 12. Although without significant differences, male tortoises had higher tick abundances than females. The interaction of tortoise sex and body condition was significantly related to tick abundance, male body condition decreased with higher tick abundance in contrast to females. Nevertheless, the interaction of body condition and reproductive stage of females was not significantly related to tick abundance. Gravid females were significantly associated with tick abundance, showing a slightly higher infestation than non-gravid females. Molecular analysis of pooled tick samples revealed the presence of Ehrlichia ewingii, Candidatus Midichloria mitochondrii, and Rickettsia africae, with a minimum infection rate of 0.61 to 1.84%. However, blood sample analysis of the tortoises was infectious agent-free, pinpointing a lack of significant health problems. Given the possible effect on the transmission of zoonotic diseases by spur-thighed tortoises associated with their frequent collection as pets, it should be surveyed to control possible human health problems. In conservation terms, as a long-lived species, the role of tick infestation in demographic traits might be included in the management and conservation programs of spur-thighed tortoises. [ABSTRACT FROM AUTHOR]

Simple Summary: Over the years, conservation physiologists have attempted to evaluate the threat factors affecting populations and manage ecosystems effectively by understanding their physiological responses. Amphibians are currently considered to be one of the most endangered vertebrate groups, and many researchers are contributing to their conservation efforts through physiological analysis. Our review aimed to examine current research trends in the conservation physiology of amphibians and identify areas for potential future studies. We categorized the 796 studies into 10 research topics. In each topic, strategies to achieve the goals of conservation physiology were identified. Additionally, we propose four comprehensive directions for future research to advance amphibian conservation physiology. Analysis of physiological responses can be used to assess population health, identify threat factors, and understand mechanisms of stress. In addition to this, conservation physiologists have sought to establish potential management strategies for environmental change and evaluate the effectiveness of conservation efforts. From past to present, the field of conservation physiology is developing in an increasingly broader context. In this review, we aim to categorize the topics covered in conservation physiology research on amphibians and present the measured physiological parameters to provide directions for future research on conservation physiology. Physiological responses of amphibians to environmental stressors are the most studied topic, but conservation physiological studies on metamorphosis, habitat loss and fragmentation, climate change, and conservation methods are relatively lacking. A number of physiological indices have been extracted to study amphibian conservation physiology, and the indices have varying strengths of correlation with each subject. Future research directions are suggested to develop a comprehensive monitoring method for amphibians, identify interactions among various stressors, establish physiological mechanisms for environmental factors, and quantify the effects of conservation activities on amphibian physiology. [ABSTRACT FROM AUTHOR]

Hybrid zones are geographic areas where individuals from distinct taxa meet, mate, and hybridize. These zones may have complex histories, but many of them originated relatively recently, during climatic oscillations in the Quaternary, following range shifts of formerly isolated, well-differentiated lineages. The Bufo bufo species group comprises four species distributed over the Western Palearctic. Whereas the contact zone between Bufo bufo and B. spinosus in western Europe has been well characterized, little is known about other species contacts. Here we focused on the contact between B. verrucosissimus and B. bufo in northeastern Türkiye, using mtDNA and microsatellite markers to describe genetic structure and patterns of admixture in the hybrid zone based on Bayesian clustering and cline analyses. Both species meet in a narrow contact zone at Rize province, with restricted introgression suggesting barriers to hybridization consistent with species status. Spatial population genetic analyses of microsatellite data pinpoint a possible enclave population of B. bufo at the Borçka district in Artvin province, isolated within the B. verrucosissimus range. The centers of the microsatellite and mtDNA-based clines are slightly displaced, with B. verrucosissimus mtDNA introgressing about 33 km W of the nuclear contact. Hybrid zone dynamics seem to be associated with range shifts mediated by Pleistocene glacial cycles and/or sex-biased dispersal. [ABSTRACT FROM AUTHOR]

Despite recent advances in the molecular knowledge of amphistome trematodes, most genera known from fish remain to be genetically characterized. This is the case for Kalitrema, a genus of the speciose family Cladorchiidae and the type of Kalitrematinae. The type and only species of this genus, Kalitrema kalitrema Travassos, 1933, was originally proposed based on two specimens found in an armored suckermouth catfish from Brazil, and its phylogenetic position has not been evaluated. In this study, paramphistomes found in Hypostomus alatus (2/9; 22.2%) and Hypostomus francisci (4/143; 2.8%) from the Paraobepa River (São Francisco River basin), Minas Gerais, Brazil, between December 2019 and November 2021, were subjected to morphological study. The parasites were identified in low intensity of infection [1.2 (1–2)] and redescribed as K. kalitrema. This species exhibits unique features such as a linguiform body with a circular ridge near the anterior end and a deep, median notch present at the posterior extremity of the body, apparently dividing the body into two lobes. A subset of specimens was further subject to phylogenetic analyses based on the most densely sampled markers, the nuclear ribosomal RNA (28S and ITS2) and mitochondrial cox1, which revealed the inclusion of K. kalitrema in a Neotropical clade of fish paramphistomes. The most comprehensive phylogenetic tree, based on the 28S dataset, confirmed K. kalitrema as an independent, early diverging lineage among Neotropical fish cladorchiids. However, the monophyly of Kalitrematinae was not sustained, given that species of the other kalitrematine genera Pseudocladorchis and Iquitostrema included in the phylogenetic analysis fell in a distinct clade with other fish cladorchiids. As a result, we propose here a narrower concept for Kalitrematinae sensu stricto, accommodating only Kalitrema (type genus) until a more natural subfamilial or familial classification is provided. [ABSTRACT FROM AUTHOR]

Mayflies (Ephemeroptera) are among the crucial water and habitat quality bioindicators. However, despite their intensive long-term use in various studies, more reliable mayfly DNA barcode data have been produced in a negligible number of countries, and only ~40% of European species had been barcoded with less than 50% of families covered. Despite being carried out in a small area, our study presents the second-most species-rich DNA reference library of mayflies from Europe and the first comprehensive view from an important biodiversity hotspot such as the Western Carpathians. Within 1153 sequences, 76 morphologically determined species were recorded and added to the Barcode of Life Data System (BOLD) database. All obtained sequences were assigned to 97 BINs, 11 of which were unique and three represented species never barcoded before. Sequences of 16 species with high intraspecific variability were divided into 40 BINs, confirming the presence of cryptic lineages. Due to the low interspecific divergence and the non-existing barcoding gap, sequences of six species were assigned to three shared BINs. Delimitation analyses resulted in 79 and 107 putative species respectively. Bayesian and maximum-likelihood phylogenies confirmed the monophyly of almost all species and complexes of cryptic taxa and proved that DNA barcoding distinguishes almost all studied mayfly species. We have shown that it is still sufficient to thoroughly investigate the fauna of a small but geographically important area to enrich global databases greatly. In particular, the insights gained here transcend the local context and may have broader implications for advancing barcoding efforts., (© 2024 John Wiley & Sons Ltd.)

Species with wide‐range distributions usually display high genetic variation. This variation can be partly explained by historical lineages that were temporally isolated from each other and are back into secondary reproductive contact, and partly by local adaptations. The smooth newt (Lissotriton vulgaris) is one of the most widely distributed amphibians species across Eurasia and forms a species complex with a partially overlapping distribution and morphology. In the present study, we explored the population genomic structure of smooth newt lineages in the Carpathian Basin (CB) relying on single‐nucleotide polymorphisms. Our dataset included new and previously published data to study the secondary contact zone between lineages in the CB and also tested for the barrier effect of rivers to gene flow between these lineages. We confirmed the presence of the South L. v. vulgaris Lineage distributed in Transdanubia and we provided new distribution records of L. v. ampelensis inhabiting the eastern territories of the CB. High genetic diversity of smooth newts was observed, especially in the North Hungarian Mountains and at the interfluves of the main rivers in the South with four distinct lineages of L. v. vulgaris and one lineage of L. v. ampelensis showing a low level of admixture with the spatially closest L. v. vulgaris lineage. Moreover, admixture detected at the interfluve of the main rivers (i.e. Danube and Tisza) suggested a secondary contact zone in the area. Finally, we found that the river Danube has a very weak effect on population divergence, while the river Tisza is a geographical barrier limiting gene flow between smooth newt lineages. As the range boundaries of L. v. vulgaris and L. v. ampelensis in the CB coincide with the river Tisza, our study underpins the influence of rivers in lineage diversification. [ABSTRACT FROM AUTHOR]

Central European water frog Pelophylax esculentus is a hybrid that lives sympatrically and forms genetic systems with one of its parental species, in this case L-E (P. lessonae - P. esculentus). Hybrids are restored in each generation due to hybridogenesis, a unique mode of hemiclonal reproduction. We investigated changes in the species composition, mate choice, and genome transmission in naturally formed pairs in the L-E hybridogenetic system from the Raków fish pond complex (SW Poland). Our observations showed that the genetic system remained the same for over a decade but the share of the taxa and the sex ratios within the species changed. The frequency of P. lessonae decreased by about 20% in favor of P. esculentus. The proportion of males to females of P. lessonae increased twofold and of P. esculentus fourfold. We also found that the stability of the L-E population was ensured by a specific female mate choice pattern with the dominance of homotypic P. lessonae and heterotypic ♀P. esculentus × ♂P. lessonae pairs (almost 90% of the pairs collected from nature). The youngest females and males that formed a successful amplexus were 3 years old. We noted a male-male competition manifested by the exclusion of young males (before the third hibernation) from the participation in the mating. An analysis of genome transmission to gametes revealed that females P. lessonae transmitted Lx, while P. esculentus transmitted the Rx and/or Lx genomes. Males of P. lessonae transmitted the Lx or Ly genomes, while P. esculentus transmitted the Lx, Ly, and/or Rx genomes. The high proportion of the L genomes transferred to gametes enables both the restoration of the parental species P. lessonae and the regular renewal of a new generation of hybrids. Significance statement: Central European water frog complex includes two species: Pelophylax ridibundus (genotype RR) and P. lessonae (LL), and their natural hybrid, P. esculentus (RL). Hybrid individuals of both sexes reproduce due to a specific mode of reproduction, i.e., hybridogenesis, in which hybrids eliminate one of the parental genomes (R or L) from the germ line prior to meiosis and transmit the other one (not recombined, i.e., clonal) to the gametes. The hybrids live in mixed populations with one of the parental species and are restored in each generation. This, in turn, allows a kind of balance to be maintained in a population due to specific mate choice and genome transmission. Our research showed that the stability of the studied population was ensured by female preference to P. lessonae and male-male competition manifested by the exclusion of males under 3 years of age from the participation in the mating. [ABSTRACT FROM AUTHOR]

Heavily urbanized areas can hinder dispersal and gene flow between amphibian populations. Given the growth potential of urbanization, it is important to examine how this specific environment shapes their genetic patterns at the local scale. The ability of the European green toad to successfully colonize large human settlements has been convincingly confirmed in the recent past, but little is known about its population genetics under these new conditions. In this study, we examined the effects of the environment on genetic variation, population structure, and the level of gene flow in populations of this amphibian in the city of Košice and the adjacent rural area (eastern Slovakia) using a set of neutral genetic markers. We found that urban populations had lower genetic variability than populations in adjacent rural areas; however, the degree of inbreeding was relatively low in all samples. Genetic differentiation was higher, and gene flow was more restricted in urban area, although geographic distances between sites were significantly less than in rural area (2–4 km versus 6–13 km). Our analyses suggested genetic isolation of urban populations at sites with less suitable habitat for green toads. In contrast, admixture of the population inhabiting the large city park, established on a former floodplain, with all rural populations was likely the result of an intense historical gene flow. The densely developed environment of the other urban sites likely presents a strong barrier to gene flow. The lack of suitable wetland habitat prior to development suggests that these sites were only recently colonized by a limited number of founders. Thus, we found differential effects of the city on the population structure of the green toad. Understanding current local genetic variation and structure is important for future conservation plans in urban environments. [ABSTRACT FROM AUTHOR]

The Karpathos water frog (Pelophylax cerigensis) is classified as Critically Endangered according to the IUCN Red List and its distribution is restricted on the island of Karpathos, South Aegean Sea. Utilizing a next generation sequencing approach, we obtained whole mitochondrial genomes of P. cerigensis to determine the species phylogenetic position within the genus Pelophylax and to clarify the phylogenetic relationship between the species' population on Karpathos Island and the population of P. cf. bedriagae from the neighboring Rhodes Island. High-throughput sequencing generated mean ± SD = 227 945 ± 18 306 reads averaged per sample. High quality reads were assembled resulting in the complete mitogenome of P. cerigensis of a total size of 17 922 bp. Mitogenome organization was similar to other Pelophylax species, comprising of 13 Protein Coding Genes (PCGs), 22 tRNAs, two rRNAs and one Control Region. Overall mean genetic distance for the 13 PCGs, within the Ranidae family, ranged from 8.01% (COIII) to 11.6% (ATP8), while ratios of synonymous and non-synonymous substitutions were <1 in all the 13 PCGs, indicating purifying selection. Maximum Likelihood and Bayesian Inference analyses based on the concatenated sequences of 13 PCGs and two rRNAs (12S and 16S), showed that individuals from Karpathos and Rhodes form a well-supported monophyletic group with low genetic distance between them (p-distance = 0.1%). Hence, the Rhodes populations seem to belong to P. cerigensis and not to P. bedriagae as formerly thought, highlighting the need for revision of P. cerigensis current conservation status as well as further examination of the P. bedriagae species group. [ABSTRACT FROM AUTHOR]

Within the Balkan Peninsula, topographic and climatic agents have promoted biodiversity and shaped the speciation history of many ectotherms. Here, we targeted an iconic European reptile, the nose‐horned viper species‐complex (Vipera ammodytes), and explored its spatial and temporal evolution. We (i) utilized genome‐wide single nucleotide polymorphisms to infer genetic structure and build a time‐calibrated species‐tree, and (ii) applied species distribution modelling with niche‐divergence tests among major phylogenomic clades. Geographically structured genetic diversity was found. Cycles of recurrent isolation and expansion during glacial–interglacial periods led to allopatric speciation and to secondary contacts and formation of multiple hybrid zones throughout the Balkan Peninsula. Deep divergence is still detected among populations separated by old and imminent biogeographical barriers (Pindos Mountain Range, the Cyclades islands, etc.), but in most cases speciation is incomplete. At the other end of the speciation continuum, we recognize two well‐differentiated lineages, currently lacking any evidence of gene flow; one is distributed in the Northwestern Balkans and the other in the Southeastern Balkans, further expanding into Asia. Despite their split 5 million years ago, there is no evidence of ecological divergence, as speciation probably occurred in niche‐pockets of analogous environments. These two lineages probably represent different species, while V. transcaucasiana does not merit species status. By comparing the genomic phylogenies to an updated mitochondrial one, we propose an evolutionary scenario that resolves all mitonuclear conflicts, according to which the history of the V. ammodytes species‐complex was shaped by complex processes, including a major event of introgressive hybridization with asymmetric mitochondrial capture. [ABSTRACT FROM AUTHOR]

In this study, we used genetic approaches to assess the conservation status of a protected amphibian species, the Macedonian crested newt, Triturus macedonicus, in Northern Pindos National Park (Epirus, Greece). Mitochondrial DNA sequences and multilocus genotypes of individuals from 38 breeding sites were used to infer their phylogenetic position and to detect and measure genetic variation patterns, population genetic structure, and levels of gene flow. The examined individuals fell within two major clades of the Macedonian crested newt phylogeny, being geographically separated by the Aoos River valley and Vikos Gorge. Both groups constitute separate gene pools, bearing private haplotypes and alleles, and the groups were found to be highly differentiated in both their mitochondrial and microsatellite markers. Thus, they meet all of the criteria needed to be characterized as evolutionary significant units (ESUs) that deserve a separate conservation status. Within each ESU, the particularly high inter-population differentiation and low migration rates imply a lack of connectivity between breeding sites and local isolation in favorable habitats. Our results can inform future programs and actions towards the conservation and management of the Macedonian crested newt in the area. [ABSTRACT FROM AUTHOR]

The knowledge of the diversity and geographic distribution of parasite species is the first step towards understanding processes of global epidemiology and species conservation. Despite recent increases in research on haemosporidian and haemogregarine parasites of reptiles and amphibians, we still know little about their diversity and parasite-host interactions, especially in the Iberian Peninsula, where a few studies have been conducted. In this study, the haemosporidian and haemogregarine diversity and phylogenetic relationships of the parasites in southwestern Iberian amphibians and reptiles were assessed using PCR approaches on blood samples of 145 individuals from five amphibian and 13 reptile species. The amphibians did not present any of both groups of parasites studied. Regarding reptiles, five Hepatozoon, one Haemogregarina, and one Haemocystidum haplotypes were found infecting four different species, revealing new host records for these parasites. Among them, we found one new Haemocystidium haplotype and three new and a previously reported Hepatozoon haplotype from a north African snake. The latter finding suggests that some Hepatozoon parasites may not be host-specific and have large geographic ranges even crossing geographical barriers. These results increased the knowledge about the geographic distribution and the number of known host species of some reptile apicomplexan parasites, highlighting the great unexplored diversity of them in this region. [ABSTRACT FROM AUTHOR]

In recent years, the influence of historical geological and climatic events on the evolution of flora and fauna in the Tibetan Plateau has been a hot research topic. The Qilian Mountain region is one of the most important sources of biodiversity on the Qinghai‐Tibet Plateau. Many species existed in the region during the Pleistocene glacial oscillation, and the complex geographical environment provided suitable conditions for the survival of local species. The shrinkage, expansion, and transfer of the distribution range and population size of species have significant effects on genetic diversity and intraspecific differentiation. To reveal the effects of geological uplift and climate oscillation on the evolution of fish populations in the Qilian Mountains, we investigated the genetic structure, phylogenetic relationship, and phylogeographical characteristics of genus Triplophysa species in the Qilian Mountains using the mitochondrial DNA gene (COI), three nuclear genes (RAG1, sRH, and Myh6) and 11 pairs of nuclear microsatellite markers. We collected 11 species of genus Triplophysa living in the Qilian Mountains, among which Triplophysa hsutschouensis and Triplophysa papillosolabiata are widely distributed in the rivers on the northern slope of the Qilian Mountains. There was a high degree of lineage differentiation among species, and the genetic diversity of endemic species was low. The different geographical groups of T. papillosolabiata presented some allogeneic adaptation and differentiation, which was closely related to the changes in the river system. Except for the population expansion event of T. hsutschouensis during the last glacial period of the Qinghai‐Tibet Plateau (0.025 MYA), the population sizes of other plateau loach species remained stable without significant population expansion. Starting from the east and west sides of the Qilian Mountains, T. hsutschouensis, and T. papillosolabiata showed two species colonization routes in opposite directions. The geological events of the uplift of the Qinghai‐Tibet Plateau and the climatic oscillation of the Quaternary glaciation had a great influence on the genetic structure of the plateau loach in the Qilian Mountains, which promoted the genetic differentiation of the plateau loach and formed some unique new species. The results of this study have important guiding significance for fish habitat protection in the Qilian Mountains. [ABSTRACT FROM AUTHOR]

Neotropical fish amphistomes represent a highly diverse group within the Paramphistomoidea, with wide distribution across major South American hydrological drainages. However, the limited molecular characterization of these taxa has impeded a comprehensive assessment of their evolutionary relationships and the systematic relevance of morphological features in classification schemes. Our study, based on the critical evaluation of the type material of both nominal species of Zonocotyle (type genus of the monotypic Zonocotylidae), and newly collected specimens of Zonocotyle bicaecata from Steindachnerina insculpta (Curimatidae) in the Upper Paraná River basin, Brazil, presents a morphological reappraisal of Z. bicaecata and provides molecular data (28S rDNA, ITS1-5.8S-ITS2 region, and COI mtDNA) to assess its phylogenetic relationships. Our phylogenetic analyses confirm this species belongs to the Paramphistomoidea. The most comprehensive analyses (based on 28S and COI) further indicate a close relationship with other fish amphistomes from the Neotropical region. Additionally, we emphasized the necessity for a new classification within Paramphistomoidea and briefly discussed the host range of Zonocotyle among curimatid fish hosts., (© 2024. The Author(s), under exclusive licence to Springer Nature B.V.)

Trematode infection of the second intermediate hosts can lead to changes in their fitness and, as a result, a change in the invasion rate of animal communities. It is especially pronounced during the invasion of parasite species that reduce activity due to the manipulation of hosts through the changes of their morphology and physiology. One of these cases is an anomaly P syndrome hotspot found in some populations of water frogs and toads in Europe caused by the trematode Strigea robusta metacercariae. The occurrence of pathogen and their participation in ecosystems are intrigues questions in the anomaly P phenomenon, as well as the role of planorbid snails that serve as the first intermediate hosts for many trematode species. Herein, we focused on trematodes spectra from planorbid snails and amphibians from the anomaly P hosts with the aim to undetected interactions between the pathways of parasites. Emerging cercariae of 6802 planorbid snails of dominant species (Planorbarius corneus, Planorbis planorbis, and Anisus spp.) were detected by both morphological and molecular methods in seven waterbodies in Privolzhskaya Lesostep Nature Reserve (Russia). A total of 95 sequences of 18 species were received, and 48 sequences were unique and did not present in any genetic databases. The 18 species of trematodes from snails and 14 species of trematodes from amphibian hosts (Pelophylax ridibundus; Ranidae; Anura) were detected. Three species (Echinostoma nasincovae, Tylodelphys circibuteonis and Australapatemon burti) was new for the trematode fauna of the Middle Volga River region and Russia as a whole. Eleven species of parasitic flatworms have amphibians in their life cycles and nine species used amphibians as metacercariae hosts: Echinostoma nasincovae, E. miyagawai, Echinoparyphium recurvatum, Tylodelphys circibuteonis, Neodiplostomum spathula, Paralepoderma cloacicola, Macrodera longicollis, Strigea robusta, and Strigea strigis. The occurrence of trematode species from planorbid mollusks and frogs were compared. [ABSTRACT FROM AUTHOR]

Isophya modestior (Orthoptera: Tettigoniidae) is a species distributed in central and south-eastern Europe, where its distribution is largely separated by two large rivers (Sava and Danube). Since previous studies on the song and morphology of the stridulatory file across its complete geographic range showed that the species is separated into two main groups, we decided to use phylogenetic and karyological analyses in order to evaluate the status of the previously analysed populations. Phylogenetic analyses showed the existence of two major clades within I. modestior with very high bootstrap values and posterior probabilities—Clade A: present on the Balkan Peninsula, Slovenia (Inner Carniola), Italy, Pannonian Serbia (Vršac Mts and Deronje) and Austria (Burgenland and Lower Austria); Clade B: present in Slovenia (Upper Carniola), Croatia and Austria (Carinthia), Pannonian Serbia (Fruška Gora Mt.) and Hungary. A comparison of chromosomes of 51 specimens revealed discrete differences between their karyotypes. The physical characteristics of the karyotypes included chromosome number (2n), sex chromosome (X) morphology and C-banding patterns. The standard chromosome complement of 50 specimens from different localities is characterized by 2 n  = 30 + X0 in males. In one male collected in Fruška Gora Mt. (Andrevlje), the chromosome number was reduced to 2 n  = 28 + neo-XY. Therefore, further cytogenetic studies involving larger samples, especially from Fruška Gora Mt. are needed in order to gain a more comprehensive view of the chromosome evolution in this group of Isophya species. [ABSTRACT FROM AUTHOR]

The geographic range of a species is crucial for obtaining information on the exact distribution of the species. The geographic data are important for delimiting distinct species or exploring the degree of differentiation among different populations of a species. The local details of species boundaries facilitate the study of the importance of phylogeographic background, secondary contacts, and hybrid zones, along with the relations between the species and its extrinsic environmental factors. In the present study, the range boundaries of Bufo bufo and Bufo verrucosissimus in the north-eastern region of Türkiye were delineated using an integrative taxonomic approach that utilized a combination of molecular and morphological data. According to the mtDNA results of the present study, B. bufo inhabits a single distribution from İyidere town to Çayeli town in Rize, while B. verrucosissimus is distributed from Şavşat town of Artvin to Ardeşen town in Rize. In addition, the two species coexist in Pazar, Hemşin, and Çamlıhemşin towns in Rize. The demographic analyses indicated a distinct population expansion for the B. verrucosissimus species after the Last Glacial Maximum, while the same did not occur for B. bufo. The univariate and multivariate statistical analyses conducted for the morphological data of the two species corroborated the presence of a putative contact zone between B. bufo and B. verrucosissimus. In summary, the present study resolved the non-distinct geographic boundaries between B. bufo and B. verrucosissimus species and also revealed the easternmost distribution of B. bufo in Türkiye. In addition, important evidence on the putative contact zone between the two species was indicated using an integrative taxonomic approach. [ABSTRACT FROM AUTHOR]

Related species often engage in abutting or overlapping contact zones with various strengths of interspecific competition. Biotic interactions such as these preclude the registration of the full profile of environmental variables that would describe the otherwise larger species ranges. Here, I advocate to forego full range species distribution modelling and instead focus on the ecography of the contact zone, for example with 'two-species distribution models' (TSDMs), in which presence data are contrasted against the background of environmental data. The newts Triturus cristatus and Triturus marmoratus meet in the west of France. A countrywide TSDM suggests that the contact zone of the species is located at a climatic gradient, in line with their north-eastern vs. south-western ranges. The species are also ecologically segregated by elevation and forestation, which is in line with a documented movement of the contact zone caused by hedgerow removal in lowland areas. Hindcasts for the Holocene suggest that the species contact zone was positioned at either the same place as at present or more to the south, depending on the amount of forestation. A forecast under climate warming predicts a fast movement to the north, but this scenario is deemed unrealistic. One reason is that recent habitat loss compromises dispersal and range expansion. Other species pairs to which TSDMs have been applied are listed for comparison. [ABSTRACT FROM AUTHOR]

With five currently recognized species that form several secondary contact zones, slow worms (Anguidae: Anguis) offer a valuable model to study the fate of evolutionary lineages in the face of hybridization and genetic introgression. The relationships between the Western Slow Worm Anguis fragilis and the Italian Slow Worm Anguis veronensis are particularly puzzling. Their respective distributions remain poorly known on the edges of their parapatric ranges, as both species lack external differentiation. Contra earlier mitochondrial phylogenies, new phylogenomic inferences have shown that A. fragilis and A. veronensis are sister taxa, thus casting doubts on their specific status. In this study, we analyze the A. fragilis / veronensis transition in southeastern France, based on one mitochondrial (ND2) and two nuclear (PRLR and HA1) genetic markers in 81 specimens from 61 localities. The ranges of A. fragilis and A. veronensis roughly extends northwest and southeast of the Rhône-Durance valleys, respectively, with clear signs of introgressive hybridization in the areas of contact (notably the eastern parts of the lower Rhône valley). Based on the three molecular markers analyzed, gene flow does not seem to reach outside the narrow hybrid zone, which likely indicates (incomplete) intrinsic reproductive isolation. Hence, we provisionally suggest maintaining A. veronensis as a separate species from A. fragilis. More generally, patterns of genetic divergence, external differentiation, and hybridization (both historical and contemporary) in Anguis ssp. supports a speciation continuum spanning from cryptic, genetically compatible alloparapatric lineages to phenotypically distinct, deeply diverged and fully reproductively isolated taxa able to coexist in sympatry. [ABSTRACT FROM AUTHOR]

Isolated distribution patches may represent local remnants of a formerly wider range or could have originated by human-mediated expansion beyond the natural range. Distinguishing between these two scenarios is not always straightforward. Northern crested newts (Triturus cristatus) in the Dutch coastal dunes are disconnected from the main species range by over 40 kilometres and whether they have been present historically is unclear. We genotyped crested newts from throughout the Netherlands for an mtDNA marker to determine the provenance of the coastal dune population. Because a closely related species, the Italian crested newt (T. carnifex), has an introduction history in the Netherlands, we also screened eight nuclear DNA SNP markers diagnostic for T. cristatus vs. T. carnifex. The crested newts from the coastal dunes carry a single T. cristatus mtDNA haplotype that naturally occurs in the south, but not the east, of the Netherlands. Therefore, we cannot distinguish if the population represents a natural distribution relict or is derived from an introduction. We find no evidence of genetic admixture with T. carnifex in the coastal dunes, but such admixture is apparent at another Dutch locality (far removed from a previously known genetically admixed population). Our study illustrates how difficult it can be to determine the origin of isolated populations. [ABSTRACT FROM AUTHOR]

Six families belonging to the Paramphistomoidea superfamily have been reported in South America, with only Zonocotylidae and Balanorchiidae being endemic. The Zonocotylidae was initially classified as Aspidogastrea and then as a paramphistomoid. This family comprises a single genus, Zononocotyle , with two species. It is primarily characterized by having an attachment organ with transverse ridges and a single testis. The placement of Zonocotylidae within Paramphistomoidea is controversial, as some researchers speculate that this genus is the most primitive member of the superfamily, while others consider it an aberrant form. The main objectives of our study were to provide the first sequences of Zonocotylidae and elucidate its phylogenetic position. We amplified the 28S gene from two parasites from Cyphocharax sp. from Punta Lara, Buenos Aires. Newly generated sequences were used to infer the phylogenetic relationships with other Paramphistomoidea species using a Bayesian approach. Zonocotylidae were clustered with Dadayiinae and Kalitrematinae (Cladorchiidae) species found in freshwater fishes from South America. Genetic analyses revealed that they formed a well-supported clade with cladorchiids in freshwater hosts from South America. However, the occurrence of genera of Cladorchiidae in North America, Middle America, Asia, and Australia suggested its polyphyletic nature and may indicate the need for the erection of new families. Other Paramphistomoidea families may also require further revision. The addition of new sequences to phylogenetic analyses along with a comprehensive and more detailed description of the genera will help resolve the relationships within this group. [ABSTRACT FROM AUTHOR]