Search

The fully aquatic Japanese giant salamander (Andrias japonicus) is a member of the Cryptobranchidae, and is currently distributed in western Japan, with other members of this group restricted to China and North America. Their feeding behaviour is characterized by a form of suction feeding that includes asymmetric movements of the jaw and hyobranchial apparatus. Previous studies on the North American species, Cryptobranchus alleganiensis, have suggested that this specialized jaw movement is produced by a flexible quadrate-articular joint combined with a loosely connected lower jaw symphysis including two small fibrocartilaginous pads. However, little is known about this feeding behaviour in the Asian species, nor have the three-dimensional asymmetric jaw movements been fully investigated in any member of Cryptobranchidae. In this study, we explore the asymmetric jaw movements in A. japonicus using three methods: (1) dissection of musculoskeletal structures; (2) filming of feeding behaviour to understand in which situations asymmetric feeding is used; (3) analysis of 3D movement of jaws and skull. In the third component, fresh (from frozen) specimens of A. japonicus were manipulated to replicate asymmetric and symmetric jaw movements, with the specimens CT scanned after each step to obtain the 3D morphology of the jaws at different positions. These positions were combined and their Euler angles from resting (closed) jaw position were calculated for asymmetric or symmetric jaw positions. Our filming revealed that asymmetric jaw movements are linked to the position of the prey in relation to the snout, with the jaw closest to the prey opening asymmetrically. Moreover, this action allows the salamander to simultaneously grasp prey in one side of the mouth while ejecting water on the other side, if the first suction attempt fails. The asymmetric jaw movements are performed mainly by rotation of the mandible about its long axis, with very limited lateral jaw movements. During asymmetric and symmetric jaw movements, the posterior ends of the maxilla and quadrate move slightly. The asymmetric jaw movements are permitted by a mobile quadrate-articular joint formed by wide, round cartilages, and by two small fibrocartilage pads within the jaw symphysis that act as cushions during jaw rotation. Some of these soft tissue structures leave traces on the jaws and skull, allowing feeding mode to be reconstructed in fossil taxa. Understanding cryptobranchid asymmetric jaw movement thus requires a comprehensive assessment of not only the symphysial morphology but also that of other cranial and hyobranchial elements., (© 2024 Anatomical Society.)

Tonic immobility is an antipredator defence in which the prey animal remains motionless after physical contact with the predator, pretending to be dead. This behaviour has been observed among a variety of taxa but has received only little attention in amphibian larvae. During our field studies with fire salamander larvae, we observed that larvae from different habitats display tonic immobility after handling. In our study site, we find larvae in pond and stream habitats, that differ in several aspects such as their stress response and their risk-taking behaviour, likely caused by the very different habitat conditions. We measured the time that the tonic immobility behaviour was displayed but found no difference between larvae from the two habitat types. Likewise, we also found no correlation between the size of the larvae and the duration of displaying the behaviour. In conclusion, we found that fire salamander larvae show tonic immobility, but found no evidence that the different habitat conditions influence the tonic immobility behaviour., Competing Interests: We declare no conflict of interest., (© 2024 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

The amphibian taxon includes three orders that present different morphological characteristics: Anura, Caudata, and Apoda. Their skin has a crucial role: it acts as an immune organ constituting a physical, chemical, immunological, and microbiological barrier to pathogen insult and conducts essential physiological processes. Amphibians have developed specialized features to protect the vulnerable skin barrier, including a glandular network beneath the skin surface that can produce antimicrobial and toxic substances, thus contributing to the defense against pathogens and predators. This study aims to characterize Langerhans cells in the skin of Lithobates catesbeianus (order: Anura; Shaw, 1802), Amphiuma means (order: Caudata; Garden, 1821), and Typhlonectes natans (order: Apoda; Fischer, 1880) with the following antibodies: Langerin/CD207 (c-type lectin), Major Histocompatibility Complex (MHC)II, and Toll-like receptor (TLR)2 (expressed by different types of DCs). Our results showed Langerhans cells positive for Langerin CD/207 in the epidermis of the three species; moreover, some antigen-presenting cells (APCs) in the connective tissue expressed TLR2 and MHCII. The distribution of the Langerhans cells is very similar in the three amphibians examined, despite their different habitats. A greater knowledge of the amphibian immune system could be useful to better understand the phylogeny of vertebrates and to safeguard amphibians from population declines. Furthermore, the similarities between amphibians' and human skin concerning immunological features may be useful in both biology and translational medicine.

Skin fungi are among the most dangerous drivers of global amphibian declines, and few mitigation strategies are known. For Batrachochytrium salamandrivorans (Chytridiomycota), available treatments rely on temperature, partially combined with antifungal drugs. We report the clearance of B. salamandrivorans in 2 urodelan species using a solely drug-based approach.

Infectious diseases are one of the main threats to biodiversity. The fungus Batrachochytrium dendrobatidis (Bd) is associated with several amphibian losses around the globe, and environmental conditions may dictate the success of pathogen spread. The Brazilian Amazon has been considered climatically unsuitable for chytrid fungus, but additional information on Bd dynamics in this ecoregion is still lacking. We sampled 462 amphibians (449 anurans, 4 caudatans and 9 caecilians), representing 57 species from the Brazilian Amazon, and quantified Bd infections using qPCR. We tested whether abiotic variables predicted the risk of Bd infections, and tested for relationships between biotic variables and Bd. Finally, we experimentally tested the effects of Bd strains CLFT 156 and CLFT 102 (from the southern and northern Atlantic Forest, respectively) on Atelopus manauensis. We detected higher Bd prevalence than those previously reported for the Brazilian Amazon, and positive individuals in all 3 orders of amphibians sampled. Both biotic and abiotic predictors were related to prevalence, and no variable explained infection load. Moreover, we detected higher Bd prevalence in forested than open areas, while the host's reproductive biology was not a factor. We detected higher mortality in the experimental group infected with CLFT 156, probably because this strain was isolated from a region characterized by discrepant climatic conditions (latitudinally more distant) when compared with the host's sampling site in Amazon. The lowland Brazilian Amazon is still underexplored and future studies targeting all amphibian orders are essential to better understand Bd infection dynamics in this region.

Amphibians are useful bioindicators for monitoring aquatic health and the influence of xenobiotics such as endocrine disrupting chemicals. Because aquatic ecosystems experience the majority of global pollution, aquatic organisms are most exposed and vulnerable to endocrine disruptors. Furthermore, penetration of endocrine disruptors into aquatic organisms especially in amphibians is even easier because of more permeable skin, resulting in high bioavailability and bioaccumulation of chemicals. One of the most potent endocrine disruptors is thiourea, which chemically blocks the synthesis of thyroid hormones and prevents metamorphosis in amphibians. We investigated the influence of thiourea on histomorphology of the thyroid gland in Triturus newts at the metamorphic stage, when thyroid hormone concentrations should reach their maximum level. Chronic exposure to thiourea induced hypertrophy and hyperplasia of follicular cells as well as a significant reduction of interstitial tissue. The intensity of the thyroglobulin immunostaining signal significantly decreases upon chronic exposure to thiourea. Successful cross-reactivity of human primary antibody in immunochemical detection of thyroglobulin in Urodela confirms potential homology in thyroglobulin structure throughout the vertebrates., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Morphological patterns are modeled by the interaction of functional, phylogenetic, ecological, and/or developmental constraints. In addition, the evolution of life cycle complexity can favor phenotypic diversity; however, the correlation between stages of development may constrain the evolution of some organs. Salamanders present microhabitat and life cycle diversity, providing an excellent framework for testing how these factors constrain phenotypic evolution. We reconstructed the morphological evolution of the terminal phalanx using a sample of 60 extinct and living species of salamanders. Using a geometric morphometric approach combined with comparative analyses, we further investigated the impact of phylogenetic, ecological, and/or life cycle factors on the shape of the terminal phalanx. We find that the phylogeny has some influence in determining the dorsal shape of the phalanges; whereas a relationship between microhabitat or life cycle and the dorsal and lateral shapes of the phalanx was not observed in the analyzed species. The allometric pattern found in the phalanx shape implies that small phalanges are more curved and with more truncated end than bigger phalanges. The evolutionary rate of phalanx shape was higher in the semiaquatic species, and the morphological disparity was significantly higher on biphasic groups. These results contradict the hypothesis that a complex life cycle constrains body shape. Finally, the phalanx shape of the salamander remains quite conserved from the Mesozoic. This configuration would allow them to occur in the different microhabitats occupied by the salamander lineages., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Amphibians undergo a variety of post-embryonic transitions (PETr) that are partly governed by thyroid hormone (TH). Transformation into a terrestrial form follows an aquatic larval stage (biphasic) or precedes hatching (direct development). Some salamanders maintain larval characteristics and an aquatic lifestyle into adulthood (paedomorphosis), which obscures the conclusion of their larval period. Paedomorphic axolotls exhibit elevated TH during early development that is concomitant with transcriptional reprogramming and limb emergence. A recent perspective suggested this cryptic TH-based PETr is uncoupled from metamorphosis in paedomorphs and concludes the larval period. This led to their question: "Are paedomorphs actual larvae?". To clarify, paedomorphs are only considered larval in form, even though they possess some actual larval characteristics. However, we strongly agree that events during larval development inform amphibian life cycle evolution. We build upon their perspective by considering the evolution of limb emergence and metamorphosis. Limbless hatchling larval salamanders are generally associated with ponds, while limbed larvae are common to streams and preceded the evolution of direct development. Permian amphibians had limbed larvae, so their PETr was likely uncoupled from metamorphosis, equivalent to most extant biphasic and paedomorphic salamanders. Coupling of these events was likely derived in frogs and direct developing salamanders., (© 2022 American Association for Anatomy.)

Background: West Siberia is a large region in North Eurasia, which harbours multiple climatic zones, landscape types and biomes. Its amphibian fauna is characterised by a combination of European and Asian species. For many species, this region is the place where the limits of their global ranges are located ( Ranatemporaria , R.amurensis , Bufotessitibundus ). West Siberia also has at least two non-native amphibian species ( Pelophylaxridibundus , Bufotesviridis ). The exact ranges and patterns of distribution of the West Siberian amphibian species are poorly studied. The mapping of species ranges is important for the development of conservation measures and monitoring of invasive species is required to investigate their impacts on the natural ecosystems., New Information: This work presents the most complete biogeographic and occurrence records database of the amphibians of West Siberia. To assemble the database, we digitised data from 190 published works, obtained data from major museum collections and from the data bank on the abundance and distribution of animals «Zoomonitor» by the Zoomonitoring laboratory of the Institute of Systematics and Ecology of Animals, Siberian Branch of Russian Academy of Sciences. The database also includes original and partly unpublished data collected by the authors from 1975 to 2021, as well as quality-assessed citizen science data from the iNaturalist portal. In total, the database contains 2530 records for 11 species of amphibians, including the locality data, the observation date (when known) and the source of the observation (at least one of the following: literature reference, museum sample ID, observer's name, iNaturalist link)., (Evgeniy Simonov, Valentina N Kuranova, Artem Lisachov, Vadim V Yartsev, Irina N Bogomolova.)

The North American newt genera Taricha and Notophthalmus (order Caudata) are well known for the combination of potent toxicity, aposematic coloration, and striking defense postures that protects these animals from predation. This suite of traits is centered around the neurotoxin tetrodotoxin, which causes paralysis and death in metazoans by disrupting the initiation and propagation of electrical signals in the nerves and muscles. Tetrodotoxin defends newts from predation across multiple life history stages and its role in generating arms-race coevolution between Taricha newts and garter snake (genus Thamnophis) predators is well studied. However, understanding the broader picture of chemical defenses in Taricha and Notophthalmus requires an expanded comprehension of the defensive chemical ecology of tetrodotoxin that includes possible coevolutionary interactions with insect egg predators, protection against parasites, as well as mimicry complexes associated with tetrodotoxin and aposematic coloration in both genera. Herein the authors review what is known about the structure, function, and pharmacology of tetrodotoxin to explore its evolution and chemical ecology in the North American newt. Focus is made specifically on the origin and possible biosynthesis of tetrodotoxin in these taxa as well as providing an expanded picture of the web of interactions that contribute to landscape level patterns of toxicity and defense in Taricha and Notophthalmus., (© 2022. The Author(s), under exclusive license to Springer Nature Switzerland AG.)

Although paedomorphosis is widespread across salamander families, only two species have ever been documented to exhibit paedomorphosis in Hynobiidae. One of these two exceptional species is Hynobius retardatus in which paedomorphosis was first reported in 1924, in specimens from Lake Kuttara in Hokkaido. This population became extinct after the last observation in 1932; since then, no paedomorphs of this species have been reported anywhere. Here, we report the rediscovery of paedomorphs of this species. Three paedomorph-like male salamanders were collected from a pond in the south Hokkaido in December 2020 and April 2021; in size, these specimens were similar to metamorphosed adults but they still displayed larval features such as external gills and a well-developed caudal fin. An artificial fertilization experiment demonstrated that they were sexually compatible with metamorphosed females, thus, confirming them to be paedomorphs. Future efforts to find additional paedomorphs in this and other populations are required to assess the prevalence of paedomorphosis in H. retardatus and to improve understanding of the ecology and evolution of paedomorphisis in Urodela., (© 2021. The Author(s).)

We present a taxonomic, spatial, and thematic overview of the current state of knowledge on helminth parasites of Mexican amphibians. Sixty-six host species have been studied so far, representing 17.5% of the amphibian species distributed in Mexico. A total of 139 nominal species of helminths - 68 platyhelminths, 62 nematodes, three acanthocephalans, three annelids (hirudineans), and three arthropods (pentastomids) - have been recorded parasitizing these hosts. Most taxa found in larval stages have not been identified at the species level. The gastrointestinal nematode Aplectana itzocanensis exhibits the broadest host range, while the bladder fluke Gorgoderina attenuata and A. itzocanensis show the widest geographic distribution. Our analysis of helminthological studies evidenced gaps and biases on research efforts that have been devoted to relatively few host species, regions, and approaches. Most helminthological records come from two species, the cane toad Rhinella marina and the Montezuma's frog Lithobates montezumae, and most studies have focused on describing the helminth fauna of a host species in a particular location or on the description of new helminth species. The highest proportion of records corresponds to the Veracruzan biogeographic province, and helminth richness is significantly correlated with host richness and with total amphibian richness by biogeographic province. Only three provinces (Yucatan Peninsula, Pacific Lowlands, and Baja Californian) have positive, yet still low helminth species discovery effort. Based on our findings, we recommend pursuing research approaches unexplored in Mexico and we provide guidelines to improve research on helminths parasitizing amphibians.

The growing availability of genetic data sets, in combination with machine learning frameworks, offers great potential to answer long-standing questions in ecology and evolution. One such question has intrigued population geneticists, biogeographers, and conservation biologists: What factors determine intraspecific genetic diversity? This question is challenging to answer because many factors may influence genetic variation, including life history traits, historical influences, and geography, and the relative importance of these factors varies across taxonomic and geographic scales. Furthermore, interpreting the influence of numerous, potentially correlated variables is difficult with traditional statistical approaches. To address these challenges, we analysed repurposed data using machine learning and investigated predictors of genetic diversity, focusing on Nearctic amphibians as a case study. We aggregated species traits, range characteristics, and >42,000 genetic sequences for 299 species using open-access scripts and various databases. After identifying important predictors of nucleotide diversity with random forest regression, we conducted follow-up analyses to examine the roles of phylogenetic history, geography, and demographic processes on intraspecific diversity. Although life history traits were not important predictors for this data set, we found significant phylogenetic signal in genetic diversity within amphibians. We also found that salamander species at northern latitudes contained low genetic diversity. Data repurposing and machine learning provide valuable tools for detecting patterns with relevance for conservation, but concerted efforts are needed to compile meaningful data sets with greater utility for understanding global biodiversity., (© 2020 John Wiley & Sons Ltd.)

Helminth infracommunities were studied at 174 sites of Latvia in seven hosts from six amphibian taxa of different taxonomical, ontogenic and ecological groups. They were described using a standard set of parasitological parameters, compared by ecological indices and linear discriminant analysis. Their species associations were identified by Kendall's rank correlation, but relationships with host size and waterbody area were analysed by zero-inflated Poisson and zero-inflated negative binomial regressions. The richest communities (25 species) were found in post-metamorphic semi-aquatic Pelophylax spp. frogs, which were dominated by trematode species of both adult and larval stages. Both larval and terrestrial hosts yielded depauperate trematode communities with accession of aquatic and soil-transmitted nematode species, respectively. Nematode loads peaked in terrestrial Bufo bufo. Helminth infracommunities suggested some differences in host microhabitat or food object selection not detected by their ecology studies. Associations were present in 96% of helminth species (on average, 7.3 associations per species) and dominated positive ones. Species richness and abundances, in most cases, were positively correlated with host size, which could be explained by increasing parasite intake rates over host ontogeny (trematode adult stages) or parasite accumulation (larval Alaria alata). Two larval diplostomid species (Strigea strigis, Tylodelphys excavata) had a negative relationship with host size, which could be caused by parasite-induced host mortality. The adult trematode abundances were higher in larger waterbodies, most likely due to their ecosystem richness, while higher larval abundances in smaller waterbodies could be caused by elevated infection rates under high host densities.

Determining the range, status, ecology and behaviour of species from areas where surveys and samplings are uncommon or difficult to conduct is a challenge, such as in the Democratic People's Republic of Korea (DPR Korea). Here, we used genetic samples, field surveys, call recordings, photographic identification and a literature review to estimate the presence, range and status of amphibians in the DPR Korea. From our combined results and based on the IUCN Red List categories and criteria, we were able to estimate the national threat levels for most species. Our results demonstrated the presence of 18 native species and the suspected presence of Karsenia koreana and two Onychodactylus species. We reported the first record for Rana uenoi in the vicinity of Pyongyang using molecular tools and similarly confirmed the presence of Dryophytes japonicus at the same location. Based on distribution and modelling, we can expect the contact zone between species within the Rana and Onychodactylus genera to be located along the Changbai Massif, a mountain range that marks a shift in ecoregions and acts as a barrier to dispersion. The species richness was higher in the lowlands and at lower latitudes, with such areas populated by up to 11 species, while more northern regions were characterised by species richness of about half of that value. The combination of ecological models and known threats resulted in the recommendation of ten species as threatened at the national level following the IUCN Red List categories and criteria. This high number of threatened species was anticipated based on the high threat level to amphibians in bordering nations and globally. While the ecology of species in the DPR Korea is still understudied, we argue that species relying on agricultural wetlands such as rice paddies are not under imminent threat due to the enduring presence of extensive agricultural landscapes with low rates of chemical use and mechanisation. The maintenance of such landscapes is a clear benefit to amphibian species, in contrast to more industrialised agricultural landscapes in neighbouring nations. In comparison, the status of species dependent on forested habitats is unclear and threat levels are likely to be higher because of deforestation, as in neighbouring nations.

Dramatic declines in amphibians worldwide highlight the need for a better understanding of diseases affecting these species. To delineate the health issues of amphibians submitted to Texas A&M University System (2016-2020), the databases were queried on the basis of defined selection criteria. A total of 502 anurans (157 frogs [44 species] and 345 toads [10 species]) and 30 caudatans (23 salamanders [6 species] and 7 newts [4 species]) were reviewed. A most likely cause of death or major pathological finding (CD-MPF) leading to euthanasia was identified in 295 (55%) Anura cases and 15 (50%) Caudata cases. Of the 532 records reviewed, anurans included 492 captive, seven free-ranging and three undetermined specimens. All caudatans were captive. The most common CD-MPF in anurans was infectious/inflammatory (228/295; 77%), involving mycobacteriosis (73/228; 32%), chlamydiosis (44/228; 19%) and mycosis (32/228; 14%). Neoplasia was less common (28/295; 9%). Infectious/inflammatory lesions (14/15; 93%) were the main CD-MPFs in caudatans. Infectious diseases are a significant threat to captive amphibians in Texas and these results may aid personnel involved in amphibian conservation programmes, veterinarians and diagnosticians., (Copyright © 2021 Elsevier Ltd. All rights reserved.)

The salamander genus Salamandra is widespread across Europe, North Africa, and the Near East and is renowned for its conspicuous and polymorphic colouration and diversity of reproductive modes. The phylogenetic relationships within the genus, and especially in the highly polymorphic species S. salamandra, have been very challenging to elucidate, leaving its real evolutionary history and classification at species and subspecies levels a topic of debate and contention. However, the distribution of diversity and species delimitation within the genus are critically important for identifying evolutionarily significant units for conservation and management, especially in light of threats posed by the pathogenic chytrid fungus Batrachochytrium salamandrivorans that is causing massive declines of S. salamandra populations in central Europe. Here, we conducted a phylogenomic analysis from across the taxonomic and geographic breadth of the genus Salamandra in its entire range. Bayesian, maximum likelihood and network-based phylogenetic analyses of up to 4905 ddRADseq-loci (294,300 nucleotides of sequence) supported the distinctiveness of all currently recognised species (Salamandra algira, S. atra, S. corsica, S. infraimmaculata, S. lanzai, and S. salamandra), and all five species for which we have multiple exemplars were confirmed as monophyletic. Within S. salamandra, two main clades can be distinguished: one clade with the Apenninic subspecies S. s. gigliolii nested within the Iberian S. s. bernardezi/fastuosa; and a second clade comprising all other Iberian, Central and East European subspecies. Our analyses revealed that some of the currently recognized subspecies of S. salamandra are paraphyletic and may require taxonomic revision, with the Central- and Eastern-European subspecies all being poorly differentiated at the analysed genomic markers. Salamandra s. longirostris - sometimes considered a separate species - was nested within S. salamandra, consistent with its subspecies status. The relationships identified within and between Salamandra species provide valuable context for future systematic and biogeographic studies, and help elucidate critical evolutionary units for conservation and taxonomy., (Copyright © 2020 Elsevier Inc. All rights reserved.)

With amphibian populations facing a multitude of threats, including habitat loss, climate change, invasive species and infectious diseases, it is important to identify valuable amphibian habitat and the imminent pressures these environments face. Between 2004 and 2019, 6 years of amphibian surveys were conducted at Greenburn, Roe and McLean lakes in the Southern Gulf Islands of British Columbia, Canada. We assessed (1) species composition and trends of native amphibians, including at-risk northern red-legged frog (Rana aurora); (2) observations of invasive American bullfrog (Lithobates catesbeianus); and (3) the efficacy of visual encounter and trapping survey methods in determining multi-species amphibian occupancy. The shallow, semi-ephemeral McLean Lake hosted more amphibian species and more breeding activity than the larger, deeper waters of Greenburn and Roe lakes. Despite multiple observations, bullfrogs have thus far not established a detectable population within these lakes, with the presence of native and introduced predators as potential contributing factors. Declining trends in occupancy of native populations of R. aurora, Pacific chorus frog (Pseudacris regilla) and rough-skinned newt (Taricha granulosa) were observed at all three lakes. Results varied within years by species and survey method, highlighting the importance of effective replication and employing complementary survey methods to optimize studies of amphibian occupancy. These observations also emphasize the value of shallow, small- to medium-sized waterbodies to native amphibian populations in the Southern Gulf Islands. As these waterbodies become increasingly threatened by global climate change and habitat degradation, the potential impacts of declining freshwater ecosystem health on amphibian populations should be considered.

Alongside exotic reptiles, amphibians, such as toads, frogs, salamanders, and newts, are nowadays considered popular pets worldwide. As reported for other exotic pet animals, amphibians are known to harbor numerous gastrointestinal parasites. Nonetheless, very little data are available on captive amphibian parasitic diseases. In this study, we applied direct saline fecal smears (DSFS) to examine in total 161 stool samples from 41 different amphibian species belonging to the orders Anura and Caudata. In addition, carbolfuchsin-smear (CFS) staining (n = 74 samples) was used to detect amphibian Cryptosporidium oocysts. Also, complete dissections of deceased amphibians (n = 107) were performed to specify parasite infections and to address parasite-associated pathogenicity. Overall, examined amphibian fecal samples contained 12 different parasite taxa. The order Rhabditida with the species Rhabdias spp. and Strongyloides spp. were the most prevalent nematode species (19.3%), followed by flagellated protozoans (8.7%), Amphibiocapillaria spp./Neocapillaria spp. (7.5%), Oswaldocruzia spp. (4.3%), Blastocystis spp. (3.1%), Cosmocerca spp. (3.1%), oxyurids (Pharyngonoidae) (3.1%), spirurids (1.2%), un-sporulated coccidian oocysts (0.6%), Tritrichomonas spp. (0.6%), Karotomorpha spp. (0.6%), and Cryptosporidium spp. (0.6%). One CFS-stained fecal sample (1.4%) was positive for Cryptosporidium oocysts. Within dissected amphibians, 31 (48.4%) of the anurans and 11 (26.2%) of the salamanders were infected with gastrointestinal parasites. One cutaneous Pseudocapillaroides xenopi infection was diagnosed in an adult African clawed frog (Xenopus laevis). Etiologically, 17 (15.9%) of them died due to severe parasitic and/or bacterial infections (e.g., Chryseobacterium indologenes, Citrobacter freudii, Sphingobacterium multivorum, Klebsiella pneumoniae). High prevalence and pathological findings of several clinical amphibian parasitoses call for more detailed investigation on gastrointestinal parasite-derived molecular mechanisms associated with detrimental lesions or even death.

The worldwide biodiversity crisis with the resulting need to increase species protection has led researchers to pursue and select survey methods that guarantee the best quality of data and produce the least negative effects on wild animals. Plethodontids are the most diverse family of salamanders; all species are very sensitive to human handling and noninvasive, but accurate, measurement methods are needed to reduce researchers' impact. Here, we tested the reliability of a noninvasive post hoc method in estimating the snout-vent length (SVL) from photographs showing salamanders' dorsal view. The correlation between the estimated snout-vent length (SVL e ) and the conventional SVL was high ( R 2 m  = .81), and no significant difference occurred between operators with different experience. Finally, we list the numerous advantages for the use of SVL e in terms of data quality and in reducing the stress caused to wild animals., Competing Interests: None declared., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Hynobius dunni is a salamander species of the genus Hynobius endemically distributed in eastern Kyushu in southwestern Japan. In this study, we determined the complete mitochondrial genome sequence and clarified the phylogenetic position of this species. The mitochondrial genome was 16,47 bp in length and encoded 13 protein, 2 ribosomal RNA, and 22 transfer RNA genes. Phylogenetic tree based on 13 protein-coding genes revealed that H. nebulosus were the most closely related species within the Hynobius species. The data identified in this study will be useful for population and conservation genetic studies of Hynobius species., (© 2020 The Author(s). Published by Informa UK Limited, trading as Taylor & Francis Group.)

While many morphological, physiological, and ecological characteristics of organisms scale with body size, some do not change under size transformation. They are called invariant. A recent study recommended five criteria for identifying invariant traits. These are based on that a trait exhibits a unimodal central tendency and varies over a limited range with body mass (type I), or that it does not vary systematically with body mass (type II). We methodologically improved these criteria and then applied them to life history traits of amphibians, Anura, Caudata (eleven traits), and reptiles (eight traits). The numbers of invariant traits identified by criteria differed across amphibian orders and between amphibians and reptiles. Reproductive output (maximum number of reproductive events per year), incubation time, length of larval period, and metamorphosis size were type I and II invariant across amphibians. In both amphibian orders, reproductive output and metamorphosis size were type I and II invariant. In Anura, incubation time and length of larval period and in Caudata, incubation time were further type II invariant. In reptiles, however, only number of clutches per year was invariant (type II). All these differences could reflect that in reptiles body size and in amphibians, Anura, and Caudata metamorphosis (neotenic species go not through it) and the trend toward independence of egg and larval development from water additionally constrained life history evolution. We further demonstrate that all invariance criteria worked for amphibian and reptilian life history traits, although we corroborated some known and identified new limitations to their application., Competing Interests: The authors declare that they have no conflict of interest., (© 2020 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Pastureland currently occupies 26% of Earth's ice-free land surface. As the global human population continues to increase and developing countries consume more protein-rich diets, the amount of land devoted to livestock grazing will only continue to rise. To mitigate the loss of global biodiversity as a consequence of the ever-expanding amount of land converted from native habitat into pastureland for livestock grazing, an understanding of how livestock impact wildlife is critical. While previous reviews have examined the impact of livestock on a wide variety of taxa, there have been no reviews examining how global livestock grazing affects amphibians. We conducted both an empirical study in south-central Florida examining the impact of cattle on amphibian communities and a quantitative literature review of similar studies on five continents. Our empirical study analyzed amphibian community responses to cattle as both a binary (presence/absence) variable, and as a continuous variable based on cow pie density. Across all analyses, we were unable to find any evidence that cattle affected the amphibian community at our study site. The literature review returned 46 papers that met our criteria for inclusion. Of these studies, 15 found positive effects of livestock on amphibians, 21 found neutral/mixed effects, and 10 found negative effects. Our quantitative analysis of these data indicates that amphibian species that historically occurred in closed-canopy habitats are generally negatively affected by livestock presence. In contrast, open-canopy amphibians are likely to experience positive effects from the presence of livestock, and these positive effects are most likely to occur in locations with cooler climates and/or greater precipitation seasonality. Collectively, our empirical work and literature review demonstrate that under the correct conditions well-managed rangelands are able to support diverse assemblages of amphibians. These rangeland ecosystems may play a critical role in protecting future amphibian biodiversity by serving as an "off-reserve" system to supplement the biodiversity conserved within traditional protected areas., (© 2019 by the Ecological Society of America.)

Copper (Cu) appears to be consistently more toxic to anuran species relative to other vertebrate taxa. There are limited Cu toxicity data for salamanders; of the few studies conducted on salamanders, most examined Cu effects on the embryonic, but not the larval, stage. We performed acute toxicity experiments, to quantify LC50s, on Harrison stage 46 larvae (free swimming hatchlings with egg yolk completely absorbed) of three ambystomatid salamander species. Each LC50 experiment used exposure concentrations of 10, 20, 30, 40, 50, and 60 µg/L with 10 replicates per concentration each containing one larva. We found very high toxicity for all species compared to previously published research on the embryo stage. Specifically, the 4-d LC50s for Ambystoma tigrinum and A. opacum were 35.3 and 18.73 µg/L, respectively. The same Cu concentrations caused similar toxicity to A. talpoideum (LC50 = 47.88 µg/L), but exposures required up to 48 d to elicit the same level of mortality. A time-to-event analysis indicated that time to mortality was significantly affected by Cu concentration. Additionally, for A. talpoideum, we observed that elevated levels of Cu decreased growth rate. Comparisons with previously reported Cu toxicity for embryos suggest that, as with fish, Cu may be more toxic to larval salamander stages than for embryos. Further, our data suggest that Cu is an important environmental contaminant that deserves increased scrutiny on the potential for population-level effects where contamination has occurred in wetlands and streams inhabited by salamanders.

Nanomaterials (NMs) have been used in a growing number of commercial products, and their rapid expansion could lead to their release into the aquatic environments. However, there is limited knowledge about the impact of NMs in the biota, especially the amphibians. The present study revised the historical use of amphibian species as a model system for nanoecotoxicological studies and summarized the data available in the scientific literature about the genotoxic, mutagenic, histopathological, embryotoxic and reproductive effects of NMs in different groups of amphibians. The interaction, bioaccumulation, mode of action (MoA) and ecotoxicity of NMs on amphibians were also revised. The nanoecotoxicological studies were conducted with 11 amphibian species, being eight species of the order Anura and three species of the order Caudata. Xenopus laevis was the most studied species. The studies were conducted mainly with inorganic NMs (72%) compared to organic ones. The nanoecotoxicity depends on NM behavior and transformation in the environment, as well as the developmental stages of amphibians. The known effects of NMs in amphibians were mainly reported with reactive oxygen species (ROS) production, oxidative stress, and genotoxic effects. Results emphasize the need for further studies testing the ecotoxicity of different NMs, concentrations and exposure periods at environmentally relevant approaches. Furthermore, standard protocols for nanoecotoxicological tests using amphibians are required. Revised data showed that amphibians are suitable organisms to assess the environmental impact of NMs and indicated significant research gaps concerning the ecotoxicity of NMs on freshwater ecosystems and recommendations for future researches., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Salamanders have some of the largest, and most variable, genome sizes among the vertebrates. Larger genomes have been associated with larger cell sizes, lower metabolic rates, and longer embryonic and larval durations in many different taxonomic groups. These life-history traits are often important for dictating fitness under different environmental conditions, suggesting that a species' genome size may have the potential to constrain its ecological distribution. We test how genome size varies with the ephemerality of larval habitat across the salamanders, predicting that species with larger genomes will be constrained to more permanent habitats that permit slower development, while species with smaller genomes will be more broadly distributed across the gradient of habitat ephemerality. We found that salamanders with larger genomes are almost exclusively associated with permanent aquatic habitats. In addition, the evolutionary transition rate between permanent and ephemeral larval habitats is much higher in salamander lineages with smaller genome sizes. These patterns suggest that genome size may act as an evolutionary constraint on the ecological habitats of salamanders, restricting those species with large genomes and slower development to habitats with permanent sources of water.

Poly- and perfluoroalkyl substances (PFAS) are ubiquitous contaminants that can bioaccumulate in aquatic taxa. Amphibians are particularly vulnerable to contaminants and sensitive to endocrine disruptors during their aquatic larval stage. However, few studies have explored PFAS uptake rates in amphibians, which is critical for designing ecotoxicology studies and assessing the potential for bioaccumulation. Uptake rates of perfluorooctane sulfonate (PFOS) and perfluorooctanoic acid (PFOA) were measured for larval northern leopard frogs (Rana pipiens), American toads (Anaxyrus americanus), and eastern tiger salamanders (Ambystoma tigrinum) during a 240-h exposure to 10 and 1000 μg/L concentrations. We measured body burden and calculated bioconcentration factor (BCF) every 48 h during the experiments. For all species and exposures, body burdens often reached steady state within 48-96 h of exposure. Steady-state body burdens for PFOA and PFOS ranged from 3819 to 16,481 ng/g dry weight (BCF = 0.46-2.5) and 6955-489,958 ng/g dry weight (47-259 BCFs), respectively. Therefore, PFAS steady state occurs rapidly in the larval amphibians we studied and particularly for PFOS. This result reflects a high potential for PFAS trophic transfer because amphibians are often low in trophic position and are important prey for many aquatic and terrestrial species., (Copyright © 2019. Published by Elsevier Inc.)

Aim: For many endemic species with limited dispersal capacities, the relationship between landscape changes and species distributions is still unclear. We characterized the population structure of the endemic ringed salamander ( Ambystoma annulatum ) across its distribution in the Central Interior Highlands (CIH) of North America, an area of high species endemism, to infer the ecological and evolutionary history of the species., Methods: We sampled 498 individuals across the species distribution and characterized the population genetic structure using nuclear microsatellite and mitochondrial DNA (mtDNA) markers., Results: Ambystoma annulatum exist in two strongly supported nuclear genetic clusters across the CIH that correspond to a northern cluster that includes the Missouri Ozark populations and a southern cluster that includes the Arkansas and Oklahoma Ozarks and the Ouachita Mountains. Our demographic models estimated that these populations diverged approximately 2,700 years ago. Pairwise estimates of genetic differentiation at microsatellite and mtDNA markers indicated limited contemporary gene flow and suggest that genetic differentiation was primarily influenced by changes in the post-Pleistocene landscape of the CIH., Main Conclusions: Both the geologic history and post-European settlement history of the CIH have influenced the population genetic structure of A. annulatum . The low mtDNA diversity suggests a retraction into and expansion out of refugial areas in the south-central Ozarks, during temperature fluctuations of the Pleistocene and Holocene epochs. Similarly, the estimated divergence time for the two nuclear clusters corresponds to changes in the post-Pleistocene landscape. More recently, decreased A. annulatum gene flow may be a result of increased habitat fragmentation and alteration post-European settlement., Competing Interests: The authors declare no conflicts of interest associated with this work., (© 2019 The Authors. Ecology and Evolution published by John Wiley & Sons Ltd.)

Patterns of phenotypic evolution can abruptly shift as species move between adaptive zones. Extant salamanders display three distinct life cycle strategies that range from aquatic to terrestrial (biphasic), to fully aquatic (paedomorphic) and to fully terrestrial (direct development). Life cycle variation is associated with changes in body form such as loss of digits, limb reduction or body elongation. However, the relationships among these traits and life cycle strategy remain unresolved. Here, we use a Bayesian modelling approach to test whether life cycle transitions by salamanders have influenced rates, optima and integration of primary locomotory structures (limbs and trunk). We show that paedomorphic salamanders have elevated rates of limb evolution with optima shifted towards smaller size and fewer digits compared to all other salamanders. Rate of hindlimb digit evolution is shown to decrease in a gradient as life cycles become more terrestrial. Paedomorphs have a higher correlation between hindlimb digit loss and increases in vertebral number, as well as reduced correlations between limb lengths. Our results support the idea that terrestrial plantigrade locomotion constrains limb evolution and, when lifted, leads to higher rates of trait diversification and shifts in optima and integration. The basic tetrapod body form of most salamanders and the independent losses of terrestrial life stages provide an important framework for understanding the evolutionary and developmental mechanisms behind major shifts in ecological zones as seen among early tetrapods during their transition from water to land., (© 2019 European Society For Evolutionary Biology. Journal of Evolutionary Biology © 2019 European Society For Evolutionary Biology.)

Amphibians are among the most highly threatened lineages, with at least 2,000 species estimated to be in danger of extinction [1, 2]. Alarmingly, another ∼2,200 species (∼25% of all ∼7,900 known species) are data deficient or not evaluated (hereinafter termed data deficient) by the International Union for Conservation of Nature (IUCN) [1]. Without an estimate of their status, data-deficient species are usually overlooked in conservation planning and resource allocation [3]. Amphibians have the highest proportion of data-deficient species of any vertebrate group [1, 4], which highlights the need to estimate their threat status considering potentially imminent extinctions. We apply a trait-based spatio-phylogenetic statistical framework [5] to predict threat status for data-deficient species. Because ecological, geographical, and evolutionary attributes increase extinction risk [6, 7], we used geographic distribution data [1, 8], phylogenetically imputed ecological traits, and an amphibian phylogeny [9] to provide initial baseline predictions. We estimate that half of the ∼2,200 data-deficient species are threatened with extinction (vulnerable, endangered, or critically endangered), primarily in the Neotropics and Southeast Asia. This increases the number of amphibian species estimated to be threatened with extinction by ∼50%. Of these, we predict that ∼500 species are endangered or critically endangered, and three may be extinct already. We highlight families that are most at risk and suggest where urgent conservation is needed to avert their loss. We show that some of the most vulnerable species may also be the most poorly known and offer an analytical framework for preliminary analysis of their threat status in the face of deficient empirical data., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

We provide a comparative population genetic study of the elusive amphibian, Proteus anguinus, by comparing the genetic diversity and divergence among 4 cave populations (96 individuals) sampled in the Dinaric Karst of Croatia. We developed 10 variable microsatellite markers using pyrosequencing and applied them to the 4 selected populations belonging to 4 different cave systems. The results showed strong genetic differentiation between the 4 caves corroborating with previous findings suggesting that Proteus might comprise several unrecognized taxa. Our results confirmed that gene flow should be high within the caves, whereas it is low between hydrographic systems since geological periods. Finally, we conclude that the high genetic subdivision suggests the necessity of treating the 4 studied Proteus populations as evolutionary significant units., (© The American Genetic Association 2018. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

We examine the phylogenetic structure and morphological differentiation within the Hynobius kimurae-H. boulengeri species complex-a widely-distributed group of stream-breeding hynobiid salamanders, inhabiting montane areas of western, central and eastern parts of Honshu Island, Japan. Phylogenetic relationships were assessed based on analyses of mitochondrial (mtDNA) and nuclear (nuDNA) gene fragments for a total of 51 samples representing 23 localities covering the entire range of the species complex. Morphological study included one-way analysis of variance (ANOVA) and principal components analysis (PCA) analyses of 26 morphometric and six meristic characters for 38 adult specimens of H. kimurae and three adult specimens of H. boulengeri . MtDNA genealogy supported monophyly of the H. kimurae-H. boulengeri complex, which is sister to all other Hynobius except H. retardatus . The complex is subdivided into three major clades, corresponding to the Eastern (Clade I) and Western (Clade II) populations of H. kimurae sensu lato, and to H. boulengeri (Clade III). Monophyly of H. kimurae sensu lato is only moderately supported by mtDNA, while nuDNA suggested that the Western form of H. kimurae is closer to H. boulengeri than to the eastern form. The time of the split of the H. kimurae-H. boulengeri complex is estimated as late Miocene and coincides with intensive crust movement in western Japan. Divergence between Clades I and II took place in early Pliocene and was likely influenced by the uplift of Central Japanese Highlands. All three clades were found to be different in a number of morphological characters, allowing us to describe the eastern form of H. kimurae as a new species, Hynobius fossigenus sp. nov ., Competing Interests: The authors declare that they have no competing interests.

Body mass is rarely recorded in amphibians, and other body measurements (e.g. snout to vent length, SVL) are generally collected instead. However, length measurements, when used as proxies of body mass in comparative analyses, are problematic if different taxa and morphotypes are included. We developed allometric relationships to derive body mass from SVL measurements. We fitted phylogenetic generalized least square models for frogs (Anura) and salamanders (Caudata) and for several families separately. We tested whether allometric relationships differed between species with different habitat preferences and between morphs in salamanders. Models were fitted with SVL-mass measurements for 88 frog and 42 salamander species. We assessed the predictive performance of the models by cross-validation. Overall, the models showed high explained variance and low forecasting errors. Models differed among semi-aquatic, terrestrial and arboreal frogs, and between paedomorphic and non-paedomorphic salamanders. Body mass estimates derived from our models allow for comparability of studies on multiple taxa and can be used for testing theories built upon evolutionary and ecological processes which are directly related to body mass., (© 2017 International Society of Zoological Sciences, Institute of Zoology/Chinese Academy of Sciences and John Wiley & Sons Australia, Ltd.)

Body elongation in vertebrates can be achieved by lengthening of the vertebrae or by an increase in their number. In salamanders, longer bodies are mostly associated with greater numbers of vertebrae in the trunk or tail region. However, studies on the relative contribution of the length of single vertebra to body elongation are lacking. In this study, we focus on evolutionary and ontogenetic changes in differentiation of the trunk vertebrae and the relative contribution of individual vertebrae to trunk lengthening in Triturus newts, a monophyletic group of salamanders that shows remarkable disparity in body shape. We compared juveniles and adults of the most elongated T. dobrogicus, which has 17 trunk vertebrae, with juveniles and adults of two closely related species (T. ivanbureschi and T. anatolicus belonging to the T. karelinii species complex) representing a stout and robust morphotype with thirteen trunk vertebrae. We show that trunk vertebrae are uniform in size at the juvenile stage of both analyzed morphotypes. In adults, the trunk vertebrae of the elongated T. dobrogicus are largely uniform, while in those of T. anatolicus, the first two vertebrae differ from the remaining trunk vertebrae. There was no difference in the relative contribution of individual vertebrae to body lengthening between species or stages. We conclude that body elongation in Triturus newts is achieved by increasing the number of vertebrae but not their length., (© 2017 Wiley Periodicals, Inc.)

The rise of high-throughput sequencing techniques provides the unprecedented opportunity to analyse controversial phylogenetic relationships in great depth, but also introduces a risk of being misinterpreted by high node support values influenced by unevenly distributed missing data or unrealistic model assumptions. Here, we use three largely independent phylogenomic data sets to reconstruct the controversial phylogeny of true salamanders of the genus Salamandra, a group of amphibians providing an intriguing model to study the evolution of aposematism and viviparity. For all six species of the genus Salamandra, and two outgroup species from its sister genus Lyciasalamandra, we used RNA sequencing (RNAseq) and restriction site associated DNA sequencing (RADseq) to obtain data for: (1) 3070 nuclear protein-coding genes from RNAseq; (2) 7440 loci obtained by RADseq; and (3) full mitochondrial genomes. The RNAseq and RADseq data sets retrieved fully congruent topologies when each of them was analyzed in a concatenation approach, with high support for: (1) S. infraimmaculata being sister group to all other Salamandra species; (2) S. algira being sister to S. salamandra; (3) these two species being the sister group to a clade containing S. atra, S. corsica and S. lanzai; and (4) the alpine species S. atra and S. lanzai being sister taxa. The phylogeny inferred from the mitochondrial genome sequences differed from these results, most notably by strongly supporting a clade containing S. atra and S. corsica as sister taxa. A different placement of S. corsica was also retrieved when analysing the RNAseq and RADseq data under species tree approaches. Closer examination of gene trees derived from RNAseq revealed that only a low number of them supported each of the alternative placements of S. atra. Furthermore, gene jackknife support for the S. atra - S. lanzai node stabilized only with very large concatenated data sets. The phylogeny of true salamanders thus provides a compelling example of how classical node support metrics such as bootstrap and Bayesian posterior probability can provide high confidence values in a phylogenomic topology even if the phylogenetic signal for some nodes is spurious, highlighting the importance of complementary approaches such as gene jackknifing. Yet, the general congruence among the topologies recovered from the RNAseq and RADseq data sets increases our confidence in the results, and validates the use of phylotranscriptomic approaches for reconstructing shallow relationships among closely related taxa. We hypothesize that the evolution of Salamandra has been characterized by episodes of introgressive hybridization, which would explain the difficulties of fully reconstructing their evolutionary relationships., (Copyright © 2017. Published by Elsevier Inc.)

Batrachochytrium salamandrivorans (Bsal) is a virulent fungal pathogen that infects salamanders. It is implicated in the recent collapse of several populations of fire salamanders in Europe. This pathogen seems much like that of its sister species, Batrachochytrium dendrobatidis (Bd), the agent responsible for anuran extinctions and extirpations worldwide, and is considered to be an emerging global threat to salamander communities. Bsal thrives at temperatures found in many mountainous regions rich in salamander species; because of this, we have screened specimens of salamanders representing 17 species inhabiting mountain ranges in three continents: The Smoky Mountains, the Swiss Alps, and the Peruvian Andes. We screened 509 salamanders, with 192 representing New World salamanders that were never tested for Bsal previously. Bsal was not detected, and Bd was mostly present at low prevalence except for one site in the Andes.

Understanding metapopulation dynamics requires knowledge about local population dynamics and movement in both space and time. Most genetic metapopulation studies use one or two study species across the same landscape to infer population dynamics; however, using multiple co-occurring species allows for testing of hypotheses related to different life history strategies. We used genetic data to study dispersal, as measured by gene flow, in three ambystomatid salamanders ( Ambystoma annulatum , A. maculatum , and A. opacum ) and the Central Newt ( Notophthalmus viridescens louisianensis ) on the same landscape in Missouri, USA. While all four salamander species are forest dependent organisms that require fishless ponds to reproduce, they differ in breeding phenology and spatial distribution on the landscape. We use these differences in life history and distribution to address the following questions: (1) Are there species-level differences in the observed patterns of genetic diversity and genetic structure? and (2) Is dispersal influenced by landscape resistance? We detected two genetic clusters in A. annulatum and A. opacum on our landscape; both species breed in the fall and larvae overwinter in ponds. In contrast, no structure was evident in A. maculatum and N. v. louisianensis , species that breed during the spring. Tests for isolation by distance were significant for the three ambystomatids but not for N. v. louisianensis . Landscape resistance also contributed to genetic differentiation for all four species. Our results suggest species-level differences in dispersal ability and breeding phenology are driving observed patterns of genetic differentiation. From an evolutionary standpoint, the observed differences in dispersal distances and genetic structure between fall breeding and spring breeding species may be a result of the trade-off between larval period length and size at metamorphosis which in turn may influence the long-term viability of the metapopulation. Thus, it is important to consider life history differences among closely related and ecologically similar species when making management decisions.

Co-existence between potentially competing newt species can lead to niche differentiation (e.g., in terms of diet shifts). This may cause adaptive responses involving changes in head shape. Here, we tested the hypothesis: the head shape of Lissotriton montandoni is different in conditions of co-occurrence with Ichthyosaura alpestris than in conditions in which other newt species are absent. We analysed images depicting head shape of specimens of I. alpestris and L. montandoni from a museum collection. All specimens of I. alpestris originated in a habitat where L. montandoni also occurred, whereas specimens of L. montandoni derived from populations that cohabited with I. alpestris and populations in which the presence of another newt species was not recorded. In each image, landmarks and semilandmarks were digitised. Females of L. montandoni from the population where I. alpestris also occurred were characterised by more massive heads and longer mouths in lateral views than females from sites where no other newt species occurred. Significant differences in head shape were also found when analysing ventral views between these species when they occupied the same habitat. We confirmed that the head shape of female L. montandoni differs between conditions of co-occurrence and absence of I. alpestris ; no differences were found for males. A differently shaped head may be an adaptation to diet; L. montandoni females with longer mouths and more robust basal parts of the head can feed on larger invertebrates and compete more effectively with I. alpestris . The co-existence of newt species should be taken into account in future ecomorphological studies.

The entire mitogenome of Tylototriton kweichowensis is 16 727 bp in length. It consists of 13 protein-coding genes (PCGS), 2 ribosomal RNA genes (rRNA), 22 transfer RNA genes (tRNA), and 1 control region (CR). Except for ND6 subunit and 8 tRNA genes are distributed on the L-strand, all the other PCGs and tRNA genes are located on the H-strand. "ATG" and "GTG" are the start codons of the PCGs, "TAA", "AGA", "TA-" and "T--" are the stop codons. Most of the tRNA genes can be folded into typical clover-leaf secondary structure. The genome of T. kweichowensis has two repeat sequences in the cob-noncoding region. Mitogenomic phylogenetic analysis (NJ tree) robustly resolved the genus-level relationship among the three genera Tylototriton, Echinotriton, and Pleurodeles, and which is congruent with the previous molecular phylogeny results.

The cutaneous microbiota plays a significant role in the biology of their vertebrate hosts, and its composition is known to be influenced both by host and environment, with captive conditions often altering alpha diversity. Here, we compare the cutaneous bacterial communities of 61 amphibians (both wild and captive) from Hiroshima, Japan, using high-throughput amplicon sequencing of a segment of the 16S rRNA gene. The majority of these samples came from a captive breeding facility at Hiroshima University where specimens from six species are maintained under highly standardized conditions for several generations. This allowed to identify host effects on the bacterial communities under near identical environmental conditions in captivity. We found the structure of the cutaneous bacterial community significantly differing between wild and captive individuals of newts, Cynops pyrrhogaster, with a higher alpha diversity found in the wild individuals. Community structure also showed distinct patterns when comparing different species of amphibians kept under highly similar conditions, revealing an intrinsic host effect. Bacterial communities of dorsal vs. ventral skin surfaces did not significantly differ in most species, but a trend of higher alpha diversity on the ventral surface was found in Oriental fire-bellied toads, Bombina orientalis. This study confirms the cutaneous microbiota of amphibians as a highly dynamic system influenced by a complex interplay of numerous factors.

The complete mitogenome of Tylototriton shanjing is 16,661 bp in length with GenBank accession number KR154461, which contains 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNA), 2 ribosomal RNA genes (rRNA), and 1 control region (CR). The overall base composition of this mitogenome is biased toward AT content at 59.45%. Most of the PCGs and tRNA genes are located on the H-strand, except for ND6 subunit gene and eight tRNA genes, which were distributed on the L-strand. The PCGs used "ATG" and "GTG" as the start codons, while "TAA", "TAG", "AGA", and "T-" are used as stop codons. Almost all tRNA genes were folded into typical cloverleaf secondary structures. The T. shanjing genome had two tandem repeat sequences in the cob-noncoding region. The mitogenomic phylogenetic analyses shows that the genera Echinotriton and Tylototriton were clustered into a strong supported monophyletic clade, which is a sister clade to the genus Pleurodeles, this confirms the previous phylogenetic results.

Tylototriton taliangensis was listed as a Near Threatened amphibian in IUCN red list. In this study, we sequenced the complete mitochondrial (mt) genome of this species (GenBank: KP979646) and found it contains 16,265 base pairs, which encode 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNA), 2 ribosomal RNA genes (rRNA) and 1 control region (CR). We also found that almost all PCGs and tRNA genes are located on the H-strand, except for ND6 subunit gene and eight tRNA genes, which were distributed on the L-strand. The PCGs used "ATG" and "GTG" as the start codon, while used four types of stop codons. Almost all tRNA genes were folded into typical cloverleaf secondary structures. The L-strand replication origin (OL) and a non-coding region were also found. The new mitogenomic phylogenetic tree confirms the reciprocally monophyly of the genus Tylototriton, Echinotriton and Pleurodeles with high bootstrap value. The present study will provide information for future studies on the conservation genetics and phylogeny of this species and its relatives.

Critical habitat for many species is often limited to occupied localities. For rare and cryptic species, or those lacking sufficient data, occupied habitats may go unrecognized, potentially hindering species recovery. Proposed critical habitat for the aquatic Jollyville Plateau salamander (Eurycea tonkawae) and two sister species were delineated based on the assumption that surface habitat is restricted to springs and excludes intervening stream reaches. To test this assumption, we performed two studies to understand aspects of individual, population, and metapopulation ecology of E. tonkawae. First, we examined movement and population demographics using capture-recapture along a spring-influenced stream reach. We then extended our investigation of stream habitat use with a study of occupancy and habitat dynamics in multiple headwater streams. Indications of extensive stream channel use based on capture-recapture results included frequent movements of >15 m, and high juvenile abundance downstream of the spring. Initial occupancy of E. tonkawae was associated with shallow depths, maidenhair fern presence and low temperature variation (indicative of groundwater influence), although many occupied sites were far from known springs. Additionally, previously dry sites were three times more likely to be colonized than wet sites. Our results indicate extensive use of stream habitats, including intermittent ones, by E. tonkawae. These areas may be important for maintaining population connectivity or even as primary habitat patches. Restricting critical habitat to occupied sites will result in a mismatch with actual habitat use, particularly when assumptions of habitat use are untested, thus limiting the potential for recovery.

A newly identified fungal pathogen, Batrachochytrium salamandrivorans(Bsal), is responsible for mass mortality events and severe population declines in European salamanders. The eastern USA has the highest diversity of salamanders in the world and the introduction of this pathogen is likely to be devastating. Although data are inevitably limited for new pathogens, disease-risk assessments use best available data to inform management decisions. Using characteristics of Bsalecology, spatial data on imports and pet trade establishments, and salamander species diversity, we identify high-risk areas with both a high likelihood of introduction and severe consequences for local salamanders. We predict that the Pacific coast, southern Appalachian Mountains and mid-Atlantic regions will have the highest relative risk from Bsal. Management of invasive pathogens becomes difficult once they are established in wildlife populations; therefore, import restrictions to limit pathogen introduction and early detection through surveillance of high-risk areas are priorities for preventing the next crisis for North American salamanders.

The order Caudata includes about 660 species and displays a variety of important developmental traits such as cleavage pattern and egg size. However, the cleavage process of tailed amphibians has never been analyzed within a phylogenetic framework. We use published data on the embryos of 36 species concerning the character of the third cleavage furrow (latitudinal, longitudinal or variable) and the magnitude of synchronous cleavage period (up to 3-4 synchronous cell divisions in the animal hemisphere or a considerably longer series of synchronous divisions followed by midblastula transition). Several species from basal caudate families Cryptobranchidae (Andrias davidianus and Cryptobranchus alleganiensis) and Hynobiidae (Onychodactylus japonicus) as well as several representatives from derived families Plethodontidae (Desmognathus fuscus and Ensatina eschscholtzii) and Proteidae (Necturus maculosus) are characterized by longitudinal furrows of the third cleavage and the loss of synchrony as early as the 8-cell stage. By contrast, many representatives of derived families Ambystomatidae and Salamandridae have latitudinal furrows of the third cleavage and extensive period of synchronous divisions. Our analysis of these ontogenetic characters mapped onto a phylogenetic tree shows that the cleavage pattern of large, yolky eggs with short series of synchronous divisions is an ancestral trait for the tailed amphibians, while the data on the orientation of third cleavage furrows seem to be ambiguous with respect to phylogeny. Nevertheless, the midblastula transition, which is characteristic of the model species Ambystoma mexicanum (Caudata) and Xenopus laevis (Anura), might have evolved convergently in these two amphibian orders.

Kokartus honorarius from the Middle Jurassic (Bathonian) of Kyrgyzstan is one of the oldest salamanders in the fossil record, characterized by a mixture of plesiomorphic morphological features and characters shared with crown-group salamanders. Here we present a detailed histological analysis of its long bones. The analysis of a growth series demonstrates a significant histological maturation during ontogeny, expressed by the progressive appearance of longitudinally oriented primary vascular canals, primary osteons, growth marks, remodelling features in primary bone tissues, as well as progressive resorption of the calcified cartilage, formation of endochondral bone and development of cartilaginous to bony trabeculae in the epiphyses. Apart from the presence of secondary osteons, the long bone histology of Kokartus is very similar to that of miniaturized temnospondyls, other Jurassic stem salamanders, miniaturized seymouriamorphs and modern crown-group salamanders. We propose that the presence of secondary osteons in Kokartus honorarius is a plesiomorphic feature, and the loss of secondary osteons in the long bones of crown-group salamanders as well as in those of miniaturized temnospondyls is the result of miniaturization processes. Hitherto, all stem salamander long bong histology (Kokartus, Marmorerpeton and 'salamander A') has been generally described as having paedomorphic features (i.e. the presence of Katschenko's Line and a layer of calcified cartilage), these taxa were thus most likely neotenic forms. The absence of clear lines of arrested growth and annuli in long bones of Kokartus honorarius suggests that the animals lived in an environment with stable local conditions., (© 2015 Anatomical Society.)

Understanding the processes that generate novel adaptive phenotypes is central to evolutionary biology. We used comparative analyses to reveal the history of tetrodotoxin (TTX) resistance in TTX-bearing salamanders. Resistance to TTX is a critical component of the ability to use TTX defensively but the origin of the TTX-bearing phenotype is unclear. Skeletal muscle of TTX-bearing salamanders (modern newts, family: Salamandridae) is unaffected by TTX at doses far in excess of those that block action potentials in muscle and nerve of other vertebrates. Skeletal muscle of non-TTX-bearing salamandrids is also resistant to TTX but at lower levels. Skeletal muscle TTX resistance in the Salamandridae results from the expression of TTX-resistant variants of the voltage-gated sodium channel NaV 1.4 (SCN4a). We identified four substitutions in the coding region of salSCN4a that are likely responsible for the TTX resistance measured in TTX-bearing salamanders and variation at one of these sites likely explains variation in TTX resistance among other lineages. Our results suggest that exaptation has played a role in the evolution of the TTX-bearing phenotype and provide empirical evidence that complex physiological adaptations can arise through the accumulation of beneficial mutations in the coding region of conserved proteins., (© 2014 The Author(s). Evolution © 2014 The Society for the Study of Evolution.)

Background: Disentangling evolutionary shifts in developmental timing (heterochony) is dependent upon accurate estimates of ancestral patterns. However, many classic assessments of heterochronic patterns predate robust phylogenetic hypotheses and methods for trait reconstruction, and therefore may have been polarized with untested 'primitive' conditions. Here we revisit the heterochronic modes of development that underlie the evolution of metamorphosis, maturation, and paedomorphosis in plethodontid salamanders. We focus on the tribe Spelerpini, which is a diverse clade that exhibits tremendous variation in timing of metamorphosis and maturation, as well as multiple independent instances of larval form paedomorphosis. Based on morphology and biogeography, early investigators concluded that the most recent common ancestors of plethodontids, and also spelerpines, were large salamanders, with very long larval periods and late maturation times. This prevailing assumption influenced subsequent heterochronic assessments, which concluded that most modern spelerpines (with shorter larval periods) were derived through multiple independent accelerations in larval development. It was also concluded that most occurrences of larval form paedomorphosis in this clade resulted from progenesis (acceleration of gonadal development relative to metamorphosis)., Results: By reconstructing the time to metamorphosis on a molecular-based phylogeny of plethodontids, we find that ancestral spelerpines likely had relatively shorter larval periods than previously proposed. Taken together with the credibility interval from our ancestral state estimation we show that very long larval periods are likely derived decelerations, only a few lineages have undergone appreciable accelerations in metamorphic timing, and the remaining taxa have lower probabilities of being different than the ancestral condition (possibly due to stasis). Reconstructing maturation age across nodes concomitant with the evolution of larval form paedomorphosis in one large radiation does not show clear evidence of progenesis, but more likely indicates a case of neoteny (delayed metamorphosis)., Conclusions: This study demonstrates cases in plethodontid salamanders where phylogenetic-based character reconstructions reject previously hypothesized ancestral life history conditions. As a result, several prior hypotheses of heterochronic evolution in this family are reversed.

The genus Salamandra represents a clade of six species of Palearctic salamanders of either contrasted black-yellow, or uniformly black coloration, known to contain steroidal alkaloid toxins in high concentrations in their skin secretions. This study reconstructs the phylogeny of the genus Salamandra based on DNA sequences of segments of 10 mitochondrial and 13 nuclear genes from 31 individual samples representing all Salamandra species and most of the commonly recognized subspecies. The concatenated analysis of the complete dataset produced a fully resolved tree with most nodes strongly supported, suggesting that a clade composed of the Alpine salamander (S. atra) and the Corsican fire salamander (S. corsica) is the sister taxon to a clade containing the remaining species, among which S. algira and S. salamandra are sister species. Separate analyses of mitochondrial and nuclear data partitions disagreed regarding basal nodes and in the position of the root but concordantly recovered the S. atra/S. corsica as well as the S. salamandra/S. algira relationship. A species-tree analysis suggested almost simultaneous temporal splits between these pairs of species, which we hypothesize was caused by vicariance events after the Messinian salinity crisis (from late Miocene to early Pliocene). A survey of toxins with combined gas chromatography/mass spectroscopy confirmed the presence of samandarine and/or samandarone steroidal alkaloids in all species of Salamandra as well as in representatives of their sister group, Lyciasalamandra. Samandarone was also detected in lower concentrations in other salamandrids including Calotriton, Euproctus, Lissotriton, and Triturus, suggesting that the presence and possible biosynthesis of this alkaloid is plesiomorphic within the Salamandridae., (Copyright © 2014 Elsevier Inc. All rights reserved.)

Larval salamanders (Lissamphibia: Caudata) are known to be effective suction feeders in their aquatic environments, although they will eventually transform into terrestrial tongue feeding adults during metamorphosis. Early tetrapods may have had a similar biphasic life cycle and this makes larval salamanders a particularly interesting model to study the anatomy, function, development, and evolution of the feeding apparatus in terrestrial vertebrates. Here, we provide a description of the muscles that are involved in the feeding strike in salamander larvae of the Hynobiidae and compare them to larvae of the paedomorphic Cryptobranchidae. We provide a functional and evolutionary interpretation for the observed muscle characters. The cranial muscles in larvae from species of the Hynobiidae and Cryptobranchidae are generally very similar. Most notable are the differences in the presence of the m. hyomandibularis, a muscle that connects the hyobranchial apparatus with the lower jaw. We found this muscle only in Onychodactylus japonicus (Hynobiidae) but not in other hynobiid or cryptobranchid salamanders. Interestingly, the m. hyomandibularis in O. japonicus originates from the ceratobranchial I and not the ceratohyal, and thus exhibits what was previously assumed to be the derived condition. Finally, we applied a biomechanical model to simulate suction feeding in larval salamanders. We provide evidence that a flattened shape of the hyobranchial apparatus in its resting position is beneficial for a fast and successful suction feeding strike., (Copyright © 2013 Wiley Periodicals, Inc.)