Marine introgressions and Andean uplift have driven diversification in neotropical Monkey tree frogs (Anura, Phyllomedusinae).

The species richness in the Neotropics has been linked to environmental heterogeneity and a complex geological history. We evaluated which biogeographic processes were associated with the diversification of Monkey tree frogs, an endemic clade from the Neotropics. We tested two competing hypotheses: the diversification of Phyllomedusinae occurred either in a "south-north" or a "north-south" direction in the Neotropics. We also hypothesized that marine introgressions and Andean uplift had a crucial role in promoting their diversification. We used 13 molecular markers in a Bayesian analysis to infer phylogenetic relationships among 57 species of Phyllomedusinae and to estimate their divergence times. We estimated ancestral ranges based on 12 biogeographic units considering the landscape modifications of the Neotropical region. We found that the Phyllomedusinae hypothetical ancestor range was probably widespread throughout South America, from Western Amazon to Southern Atlantic Forest, at 29.5 Mya. The Phyllomedusines' ancestor must have initially diverged through vicariance, generally followed by jump-dispersals and sympatric speciation. Dispersal among areas occurred mostly from Western Amazonia towards Northern Andes and the South American diagonal of dry landscapes, a divergent pattern from both "south-north" and "north-south" diversification hypotheses. Our results revealed a complex diversification process of Monkey tree frogs, occurring simultaneously with the orogeny of Northern Andes and the South American marine introgressions in the last 30 million years.

Immunohistochemical Characterization of Langerhans Cells in the Skin of Three Amphibian Species.

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.

Vocal repertoire of Microhyla nilphamariensis from Delhi and comparison with closely related M. ornata populations from the western coast of India and Sri Lanka.

Advertisement calls in frogs have evolved to be species-specific signals of recognition and are therefore considered an essential component of integrative taxonomic approaches to identify species and delineate their distribution range. The species rich genus Microhyla is a particularly challenging group for species identification, discovery and conservation management due to the small size, conserved morphology and wide distribution of its members, necessitating the need for a thorough description of their vocalization. In this study, we provide quantitative description of the vocal behaviour of Microhyla nilphamariensis, a widely distributed south Asian species, from Delhi, India, based on call recordings of 18 individuals and assessment of 21 call properties. Based on the properties measured acrossed 360 calls, we find that a typical advertisement call of M. nilphamariensis lasts for 393.5 ±  57.5 ms, has 17 pulses on average and produce pulses at rate of 39 pulses/s. The overall call dominant frequency was found to be 2.8 KHz and the call spectrum consisted of two dominant frequency peaks centered at 1.6 KHz and 3.6 KHz, ranging between 1.5-4.1 KHz. Apart from its typical advertisement call, our study also reveals the presence of three 'rare' call types, previously unreported in this species. We describe variability in call properties and discuss their relation to body size and temperature. We found that overall dominant frequency 1 (spectral property) was found to be correlated with body size, while first pulse period (temporal property) was found to be correlated with temperature. Further, we compare the vocal repertoire of M. nilphamariensis with that of the congener Microhyla ornata from the western coast of India and Sri Lanka and also compare the call properties of these two populations of M. ornata to investigate intra-specific call variation. We find statistically significant differentiation in their acoustic repertoire in both cases. Based on 18 call properties (out of 20), individuals of each locality clearly segregate on PCA factor plane forming separate groups. Discriminant function analysis (DFA) using PCA factors shows 100% classification success with individuals of each locality getting classified to a discrete group. This confirms significant acoustic differentiation between these species as well as between geographically distant conspecifics. The data generated in this study will be useful for comparative bioacoustic analysis of Microhyla species and can be utilized to monitor populations and devise conservation management plan for threatened species in this group.

Tonic immobility behaviour does not differ between fire salamander larvae from ponds and streams.

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.

Diversity and Evolution of Frog Visual Opsins: Spectral Tuning and Adaptation to Distinct Light Environments.

Visual systems adapt to different light environments through several avenues including optical changes to the eye and neurological changes in how light signals are processed and interpreted. Spectral sensitivity can evolve via changes to visual pigments housed in the retinal photoreceptors through gene duplication and loss, differential and coexpression, and sequence evolution. Frogs provide an excellent, yet understudied, system for visual evolution research due to their diversity of ecologies (including biphasic aquatic-terrestrial life cycles) that we hypothesize imposed different selective pressures leading to adaptive evolution of the visual system, notably the opsins that encode the protein component of the visual pigments responsible for the first step in visual perception. Here, we analyze the diversity and evolution of visual opsin genes from 93 new eye transcriptomes plus published data for a combined dataset spanning 122 frog species and 34 families. We find that most species express the four visual opsins previously identified in frogs but show evidence for gene loss in two lineages. Further, we present evidence of positive selection in three opsins and shifts in selective pressures associated with differences in habitat and life history, but not activity pattern. We identify substantial novel variation in the visual opsins and, using microspectrophotometry, find highly variable spectral sensitivities, expanding known ranges for all frog visual pigments. Mutations at spectral-tuning sites only partially account for this variation, suggesting that frogs have used tuning pathways that are unique among vertebrates. These results support the hypothesis of adaptive evolution in photoreceptor physiology across the frog tree of life in response to varying environmental and ecological factors and further our growing understanding of vertebrate visual evolution.

Multifunctional host-defense peptides isolated from skin secretions of the banana tree dwelling frog Boana platanera (Hylidae; Hylinae).

Five host-defense peptides (figainin 2PL, hylin PL, raniseptin PL, plasticin PL, and peptide YL) were isolated from norepinephrine-stimulated skin secretions of the banana tree dwelling frog Boana platanera (Hylidae; Hylinae) collected in Trinidad. Raniseptin PL (GVFDTVKKIGKAVGKFALGVAKNYLNS.NH2) and figainin 2PL (FLGTVLKLGKAIAKTVVPMLTNAMQPKQ. NH2) showed potent and rapid bactericidal activity against a range of clinically relevant Gram-positive and Gram-negative ESKAPE + pathogens and Clostridioides difficile. The peptides also showed potent cytotoxic activity (LC50 values < 30 µM) against A549, MDA-MB-231 and HT29 human tumor-derived cell lines but appreciably lower hemolytic activity against mouse erythrocytes (LC50 = 262 ± 14 µM for raniseptin PL and 157 ± 16 µM for figainin 2PL). Hylin PL (FLGLIPALAGAIGNLIK.NH2) showed relatively weak activity against microorganisms but was more hemolytic. The glycine-leucine-rich peptide with structural similarity to the plasticins (GLLSTVGGLVGGLLNNLGL.NH2) and the non-cytotoxic peptide YL (YVPGVIESLL.NH2) lacked antimicrobial and cytotoxic activities. Hylin PL, raniseptinPL and peptide YL stimulated the rate of release of insulin from BRIN-BD11 clonal ß-cells at concentrations ≥100 nM. Peptide YL was the most effective (2.3-fold increase compared with basal rate at 1 µM concentration) and may represent a template for the design of a new class of incretin-based anti-diabetic drugs.

Interspecific cytogenomic comparison reveals a potential chromosomal centromeric marker in Proceratophrys frog species.

Among the repetitive elements, satellite DNA (SatDNA) emerges as extensive arrays of highly similar tandemly repeated units, spanning megabases in length. Given that the satDNA PboSat01-176, previously characterized in P. boiei, prompted our interest for having a high abundance in P. boiei and potential for centromeric satellite, here, we employed various approaches, including low coverage genome sequencing, followed by computational analysis and chromosomal localization techniques in four Proceratophrys species and, investigating the genomic presence and sharing, as well as its potential for chromosomal centromere marker in Proceratophrys frog species. Our findings demonstrate that PboSat01-176 exhibits high abundance across all four Proceratophrys species, displaying distinct characteristics that establish it as the predominant repetitive DNA element in these species. The satellite DNA is prominently clustered in the peri/centromeric region of the chromosomes, particularly in the heterochromatic regions. The widespread presence of PboSat01-176 in closely related Proceratophrys species reinforces the validity of the library hypothesis for repetitive sequences. Thus, this study highlighted the utility of the satDNA family PboSat01-176 as a reliable centromeric marker in Proceratophrys species, with potential to be applied in other species of anuran amphibians. The observed sharing and maintenance of this sequence within the genus suggest possibilities for future research, particularly through expanded sampling to elucidate parameters that underlie the library hypothesis and the evolutionary dynamics of satDNA sequences.

Manual for the study of tapeworms (Cestoda) parasitic in ray-finned fish, amphibians and reptiles.

Based on long-term and often frustrating experiences with the poor quality of tapeworms (Cestoda) collected throughout the world for taxonomic and phylogenetic studies, and considering the increasing obstacles to obtaining new material, a simple, easy-to-use and illustrated methodological guide (manual) is provided. It focusses mainly on key steps in examining hosts, collecting cestodes from poikilothermous vertebrates except elasmobranchs, i.e., from ray-finned fish (Actinopterygii), amphibians and 'reptiles' (a paraphyletic group comprising all sauropsids except birds), and fixing them for subsequent morphological and molecular study. It is proposed that the following methodological points should be followed: (i) ideally only freshly euthanised hosts (not previously frozen) should be used for parasitological examination; (ii) hosts examined should be documented by photographs; host tissue should also be preserved for future genotyping if necessary; (iii) tapeworms should be detached carefully to keep the scolex intact and properly cleaned before fixation; (iv) a small piece of cestode tissue should be always preserved in molecular grade ethanol for DNA sequencing; (v) tapeworms should be fixed as quickly as possible after collecting them and while they are still alive, always using hot (heated) fixatives; this prevents unnatural contraction or deformation and ensures uniform fixation; (vi) each sample (vial) should be properly labelled (a unique code should be given to every cestode sample); (vii) vouchers of sequenced specimens (hologenophores or paragenophores) should always be preserved for identification, and deposited in internationally recognised collections. It is hoped that this guide helps researchers and students to properly process valuable material of cestodes to make it suitable for reliable identification including genotyping and comparative anatomy, which is a prerequisite for any subsequent ecological, biogeographical, phylogenetic life cycle or molecular study.

Morbidity in California giant salamander (Dicamptodon ensatus Eschscholtz, 1833) caused by Euryhelmis sp. Poche, 1926 (Trematoda: Heterophyiidae).

In the fall of 2021, California Department of Fish and Wildlife reported larval and adult California giant salamanders (Dicamptodon ensatus Eschscholtz, 1833) with skin lesions at multiple creeks in Santa Clara and Santa Cruz Counties, California, USA. Field signs in both stages included rough, lumpy textured skin, and larvae with tails that were disproportionately long, flat, wavy, and flaccid. Presence of large-bodied larvae suggested delayed metamorphosis, with some larvae having cloudy eyes and suspected blindness. To determine the cause of the disease, three first-of-the-year salamanders from one location were collected, euthanized with 20% benzocaine, and submitted for necropsy to the U.S. Geological Survey, National Wildlife Health Center. Upon gross examination, all salamanders were emaciated with no internal fat stores, and had multiple pinpoint to 1.5-mm diameter raised nodules in the skin over the body, including the head, gills, dorsum, ventrum, all four limbs, and the tail; one also had nodules in the oral cavity and tongue. Histologically all salamanders had multiple encysted metacercariae in the dermis, subcutis, and skeletal muscles of the head, body, and tail that were often associated with granulomatous and granulocytic inflammation and edema. A small number of encysted metacercariae or empty cysts were present in the gills with minimal inflammation, and rarely in the kidney with no associated inflammation. Morphology of live metacercariae (Trematoda: Heterophyiidae), and sequencing of the 28S rRNA gene identified a species of Euryhelmis (Poche, 1926). Artificial digestion of a 1.65 g, decapitated, eviscerated carcass yielded 773 metacercariae, all of similar size and morphology as the live specimens. Based on these findings, the poor body condition of these salamanders was concluded to be due to heavy parasite burden. Environmental factors such as drought, increased temperature, and overcrowded conditions may be exacerbating parasite infections in these populations of salamander.

Chytridiomycosis causes high amphibian mortality prior to the completion of metamorphosis.

Amphibian populations are undergoing extensive declines globally. The fungal disease chytridiomycosis, caused by the pathogenic fungus Batrachochytrium dendrobatidis (Bd), is a primary contributor to these declines. The amphibian metamorphic stages (Gosner stages 42-46) are particularly vulnerable to a range of stressors, including Bd. Despite this, studies that explicitly examine host response to chytridiomycosis throughout the metamorphic stages are lacking. We aimed to determine how Bd exposure during the larval stages impacts metamorphic development and infection progression in the endangered Fleay's barred frog (Mixophyes fleayi). We exposed M. fleayi to Bd during pro-metamorphosis (Gosner stages 35-38) and monitored infection dynamics throughout metamorphosis. We took weekly morphological measurements (weight, total body length, snout-vent-length and Gosner stage) and quantified Bd load using qPCR. While we observed minimal impact of Bd infection on animal growth and development, Bd load varied throughout ontogeny, with an infection load plateau during the tadpole stages (Gosner stages 35-41) and temporary infection clearance at Gosner stage 42. Bd load increased exponentially between Gosner stages 42 and 45, with most exposed animals becoming moribund at Gosner stage 45, prior to the completion of metamorphosis. There was variability in infection outcome of exposed individuals, with a subgroup of animals (n = 5/29) apparently clearing their infection while the majority (n = 21/29) became moribund with high infection burdens. This study demonstrates the role that metamorphic restructuring plays in shaping Bd infection dynamics and raises the concern that substantial Bd-associated mortality could be overlooked in the field due to the often cryptic nature of these latter metamorphic stages. We recommend future studies that directly examine the host immune response to Bd infection throughout metamorphosis, incorporating histological and molecular methods to elucidate the mechanisms responsible for the observed trends.

First documentation on herpetofauna diversity in Gunung Belumut Amenity Forest, Peninsular Malaysia: Implications for conservation in an Environmentally Sensitive Area (ESA).

Malaysia is blessed with lush tropical rainforests that harbour an exceptional diversity of amphibians and reptiles. However, compared to other animal groups, amphibians and reptiles have received limited attention in research, despite their ecological significance. With amphibians and reptile species having been declining rapidly due to anthropogenic activities, there is a pressing need to conserve these species and their habitats. Environmentally Sensitive Areas (ESAs) are designated regions that are beneficial due to their critical role in providing essential ecosystem services and serving as repositories of biodiversity. Nonetheless, the classification of ESAs in Malaysia lacks biological elements and only focuses on physical attributes. To enhance the current ESA classification framework by integrating biological components, there is an urgent need to obtain information on diversity and habitat in Malaysia. Therefore, the objectives of this study were twofold: to determine the diversity of amphibians and reptiles in Gunung Belumut Amenity Forest and to conduct a comparative analysis between the herpetofauna assemblages in Gunung Belumut with other forest reserves in Peninsular Malaysia. The survey was carried out between March and June 2022, with additional sampling conducted in February 2023. The Visual Encounter Survey (VES) and pitfall trap methods were employed to survey the herpetofauna species, focusing on both aquatic and terrestrial habitats within the study area. A total of 210 individuals representing 38 species of herpetofauna were recorded, comprising 18 amphibian and 20 reptile species. Amongst the observed species, Limnonectesblythii was the most frequently encountered amphibian, with 59 individuals observed, while the dominant reptile species was Cyrtodactylusconsobrinus, represented by eight individuals. This pioneering study serves as a vital baseline documentation of the amphibian and reptile assemblages in Gunung Belumut Amenity Forest. It provides valuable information for identifying extant herpetofauna species, including those of potential conservation concern or rarity. These findings contribute to ongoing conservation efforts dedicated to the preservation of herpetofauna within the region. By understanding the diversity and distribution patterns of amphibians and reptiles in Gunung Belumut, effective conservation strategies can be developed to protect these species and their habitats.

Diversification of the terrestrial frog genus Anomaloglossus (Anura, Aromobatidae) in the Guiana Shield proceeded from highlands to lowlands, with successive loss and reacquisition of endotrophy.

Two main landscapes emerge from the Guiana Shield: the highlands to the west called the Pantepui region and the Amazonian lowlands to the east, both harbouring numerous endemic species. With 32 currently recognized species, the genus Anomaloglossus stands out among Neotropical frogs as one that diversified only within the Guiana Shield both in the highlands and the lowlands. We present a time-calibrated phylogeny obtained by using combined mitogenomic and nuclear DNA, which suggests that the genus originates from Pantepui where extant lineages started diversifying around 21 Ma, and subsequently (ca. 17 Ma) dispersed during the Miocene Climatic Optimum to the lowlands of the eastern Guiana Shield where the ability to produce endotrophic tadpoles evolved. Further diversification within the lowlands in the A. stepheni group notably led to an evolutionary reversal toward exotrophy in one species group during the late Miocene, followed by reacquisition of endotrophy during the Pleistocene. These successive shifts of reproductive mode seem to have accompanied climatic oscillations. Long dry periods might have triggered evolution of exotrophy, whereas wetter climates favoured endotrophic forms, enabling colonization of terrestrial habitats distant from water. Acquisition, loss, and reacquisition of endotrophy makes Anomaloglossus unique among frogs and may largely explain the current species diversity. The micro evolutionary processes involved in these rapid shifts of reproductive mode remain to be revealed.

The cardiorespiratory system of miniature frogs.

Anurans of the genus Brachycephalus are among the smallest vertebrates in the world, due to an extreme process of miniaturization. As an example of this process, Brachycephalus species show loss of fingers, loss of the eardrum and middle ear, bone fusions, and the presence of paravertebral plates and parotic plaque. However, no studies addressing the consequences of miniaturization on internal organs, such as the lungs and heart, are currently available. Thus, this study aimed to investigate if overall small body size has affected the cardiorespiratory system. We investigated, via dissections, individuals of four Brachycephaloidea species: Brachycephalus rotenbergae, B. pitanga, Eleutherodactylus johnstonei, and Ischnocnema parva. We observed that B. rotenbergae and B. pitanga present a reduction of the atrial septum and absence of the carotid body. On the other hand, despite being a member of the sister genus to Brachycephalus (both genera belong to the Brachycephalidae), individuals of Ischnocnema present a heart with a complete septum and carotid body; this is also observed in E. johnstonei (Eleutherodactylidae). We observed that B. rotenbergae and B. pitanga have thin skin with a one to two cell thick germ layer, and their lungs likely exhibit lower blood supply when compared to individuals of the E. johnstonei and I. parva species. Based on the observed structures, we suggest that in species of Brachycephalus, respiration is performed mainly through the skin, and their lungs may have a reduced respiratory function.

Cellular and Molecular Triggers of Retinal Regeneration in Amphibians.

Understanding the mechanisms triggering the initiation of retinal regeneration in amphibians may advance the quest for prevention and treatment options for degenerating human retina diseases. Natural retinal regeneration in amphibians requires two cell sources, namely retinal pigment epithelium (RPE) and ciliary marginal zone. The disruption of RPE interaction with photoreceptors through surgery or injury triggers local and systemic responses for retinal protection. In mammals, disease-induced damage to the retina results in the shutdown of the function, cellular or oxidative stress, pronounced immune response, cell death and retinal degeneration. In contrast to retinal pathology in mammals, regenerative responses in amphibians have taxon-specific features ensuring efficient regeneration. These include rapid hemostasis, the recruitment of cells and factors of endogenous defense systems, activities of the immature immune system, high cell viability, and the efficiency of the extracellular matrix, cytoskeleton, and cell surface remodeling. These reactions are controlled by specific signaling pathways, transcription factors, and the epigenome, which are insufficiently studied. This review provides a summary of the mechanisms initiating retinal regeneration in amphibians and reveals its features collectively directed at recruiting universal responses to trauma to activate the cell sources of retinal regeneration. This study of the integrated molecular network of these processes is a prospect for future research in demand biomedicine.

Catalogue of type specimens deposited in the Herpetology Collection of the Natural History Museum Gustavo Orcés V. at Escuela Politécnica Nacional (Ecuador).

Background: The Herpetology Collection of the Natural History Museum Gustavo Orcés V. at Escuela Politécnica Nacional (MEPN-H) in Quito maintains more than sixteen thousand curated specimens and it comprises Ecuador ´s second largest collection of herps. New information: The Collection contains 193 type specimens: 14 holotypes, 34 paratopotypes and 145 paratypes, which correspond to 10 families, 17 genera and 32 species. The collection of type specimens is particularly important in the genera Atelopus and Pristimantis in amphibians and the genera Atractus and Enyaloides in reptiles. An assessment of the geographic distribution showed that collection sites of type specimens are clustered towards the south of Ecuador in the provinces of Zamora Chinchipe, Morona Santiago and Pastaza in the Amazon Region; and in the provinces of Carchi and Azuay in the Andes. The collection of type specimens dates from 1955 to 2013, comprising an invaluable source of historical biodiversity data.

Reed bamboos drive skull shape evolution in bush frogs of the Western Ghats, Peninsular India.

Reed bamboo is a major ecological and economic resource for many animals, including humans. Nonetheless, the influence of this plant's evolutionary role on the morphology of animal species remains unexplored. Here, we investigated the significance of bamboo habitats as ecological opportunities in shaping the skull morphology of bush frogs (Raorchestes) from the Western Ghats, Peninsular India. We applied a three-dimensional (3D) geometric morphometric approach to capture the skull shape of 55 species of bush frogs. We visualized the skull shape variables in phylomorphospace with principal component analysis and performed phylogenetic generalized least-squares analysis to assess the impact of cranial size (evolutionary allometry) and habitat (bamboo or non-bamboo) on cranial shape. We quantified the morphological disparity between bamboo and non-bamboo bush frogs' skull shape, and employed RRphylo, a phylogenetic ridge regression method, to access the evolutionary rate and rate shifts of skull shape change. The phylomorphospace delineated bamboo and non-bamboo bush frogs. While cranial shape exhibited a significant but smaller association with size, its association with habitat type was non-significant. We detected, however, significant differences in skull shape between the two frog groups, with bamboo frogs showing higher morphological disparity and a remarkable shift in the evolutionary rate of skull shape diversification. These findings underscore the role of reed bamboo in the evolution of skull shape in the radiation of frogs, endemic to the Western Ghats. We demonstrate that the association between the members of two distinct endemic clades (bamboo reeds and bamboo frogs) is the outcome of a deep-time ecological opportunity that dates back to the Miocene.

A geometric morphometric analysis of variation in Australian frog ilia and taxonomic interpretations.

Anurans including frogs and toads exhibit an ilium that is often regarded as taxonomically diagnostic. The ilium, one of the three paired bones that make up the pelvic girdle, has been important in the fossil record for identifying anuran morphotypes. Osteological collections for Australian frogs are rare in herpetological museums, and skeletonizing whole-bodied specimens requires destroying soft tissue morphology which is valuable to anuran specialists working on living species. Computed tomography scans provide the opportunity to study anuran osteology without the loss of soft tissues. Our study, based on microcomputed tomography scans of extant Australian frogs from the public repository Morphosource and from museum collections focuses on the morphological differences between Australian frogs at the familial and generic levels using geometric morphometrics to compare the diagnostic shape of the ilium. Principal component analysis (PCA) and canonical variate analysis (CVA) were conducted to assess differences in the ilium between supraspecific groups of Australian frogs. The canonical variates analysis accurately predicted group membership (i.e., the correct family) with up to 76.2% success for cross-validated predictions and 100% of original group predictions. While the sample was limited to familial and generic level analyses, our research shows that ilial morphology in Australian frogs is taxonomically informative. This research provides a guide for identifying Australian anurans, including fossils, as well as new information relevant to considerations about their phylogenetic relationships, and the potential use of the fossil record to enhance efforts to conserve threatened living frog species.

Life History Traits and Longevity of the Invasive Asian Common Toad Duttaphrynus melanostictus (Schneider, 1799) in Madagascar.

We analyzed the body length, age structure, and age at sexual maturity of the invasive Asian common toad Duttaphrynus melanostictus from different sites in Toamasina, east Madagascar. We used skeletochronology as a proxy for age estimation, while gonads were histologically analyzed to determine the age of sexual maturity. The analysis of pooled age data from three sites investigated in 2016 showed that both sexes were larger, although not older, than those of native populations. For the individuals from Madagascar, the males were significantly smaller and younger (mean ± SD, SVL: 71.4 ± 1.6 mm; age: 1.8 ± 0.7 years) than the females (SVL: 78.42 ± 1.9 mm; age: 2.7 ± 1.3 years), when the data were pooled, but when the data were analyzed separately for each of the three sites, similar results were obtained only for one site. The oldest recorded male and female were 3 and 6 years old, respectively. Gonadal histology showed that the males and females reach sexual maturity after the first and second years of age, respectively. Further studies are needed to understand if the larger size and faster growth rates observed in the invasive population of D. melanostictus in Madagascar are a consequence of more favorable environmental conditions with respect to the native range (e.g., the availability of larger trophic niches, a lack of competitors, and lower predatory pressure), and we suggest to extend the monitoring of these life history traits to understand how they might influence the invasion.

Thyroid and reproduction in amphibians and reptiles.

The relationship between the thyroid gland and reproduction in amphibians and reptiles has been studied for more than 100 years. Most studies suggest a positive involvement of thyroid hormones with some aspects of reproduction, but some studies support a negative role for thyroid hormones at certain life stages. Comprehensive studies of gene activation/suppression by thyroid hormones and their absence at various levels of the hypothalamo-pituitary-gonadal axis coupled with observations of adrenocorticoid activity, reproductive performance, and metabolic involvement are needed to understand this complex relationship.

Modeling the Distribution of the Chytrid Fungus Batrachochytrium dendrobatidis with Special Reference to Ukraine.

Amphibians are the most threatened group of vertebrates. While habitat loss poses the greatest threat to amphibians, a spreading fungal disease caused by Batrachochytrium dendrobatidis Longcore, Pessier & D.K. Nichols 1999 (Bd) is seriously affecting an increasing number of species. Although Bd is widely prevalent, there are identifiable heterogeneities in the pathogen's distribution that are linked to environmental parameters. Our objective was to identify conditions that affect the geographic distribution of this pathogen using species distribution models (SDMs) with a special focus on Eastern Europe. SDMs can help identify hotspots for future outbreaks of Bd but perhaps more importantly identify locations that may be environmental refuges ("coldspots") from infection. In general, climate is considered a major factor driving amphibian disease dynamics, but temperature in particular has received increased attention. Here, 42 environmental raster layers containing data on climate, soil, and human impact were used. The mean annual temperature range (or 'continentality') was found to have the strongest constraint on the geographic distribution of this pathogen. The modeling allowed to distinguish presumable locations that may be environmental refuges from infection and set up a framework to guide future search (sampling) of chytridiomycosis in Eastern Europe.