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How do chemically defended animals resist their own toxins? This intriguing question on the concept of autotoxicity is at the heart of how species interactions evolve. In this issue of Molecular Ecology (Molecular Ecology, 2024, 33), Bodawatta and colleagues report on how Papua New Guinean birds coopted deadly neurotoxins to create lethal mantles that protect against predators and parasites. Combining chemical screening of the plumage of a diverse collection of passerine birds with genome sequencing, the researchers unlocked a deeper understanding of how some birds sequester deadly batrachotoxin (BTX) from their food without poisoning themselves. They identified that birds impervious to BTX bear amino acid substitutions in the toxin-binding site of the voltage-gated sodium channel Nav1.4, whose function is essential for proper contraction and relaxation of vertebrate muscles. Comparative genetic and molecular docking analyses show that several of the substitutions associated with insensitivity to BTX may have become prevalent among toxic birds through positive selection. Intriguingly, poison dart frogs that also co-opted BTX in their lethal mantles were found to harbour similar toxin insensitivity substitutions in their Nav1.4 channels. Taken together, this sets up a powerful model system for studying the mechanisms behind convergent molecular evolution and how it may drive biological diversity.

Predator-prey interactions have been suggested as drivers of diversity in different lineages, and the presence of anti-predator defences in some clades is linked to higher rates of diversification. Warning signals are some of the most widespread defences in the animal world, and there is evidence of higher diversification rates in aposematic lineages. The mechanisms behind such species richness, however, are still unclear. Here, we test whether lineages that use aposematism as anti-predator defence exhibit higher levels of genetic differentiation between populations, leading to increased opportunities for divergence. We collated from the literature more than 3000 pairwise genetic differentiation values across more than 700 populations from over 60 amphibian species. We find evidence that over short geographical distances, populations of species of aposematic lineages exhibit greater genetic divergence relative to species that are not aposematic. Our results support a scenario where the use of warning signals could restrict gene flow, and suggest that anti-predator defences could impact divergence between populations and potentially have effects at a macro-evolutionary scale.

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.

Frogs are an ecologically diverse and phylogenetically ancient group of anuran amphibians that include important vertebrate cell and developmental model systems, notably the genus Xenopus. Here we report a high-quality reference genome sequence for the western clawed frog, Xenopus tropicalis, along with draft chromosome-scale sequences of three distantly related emerging model frog species, Eleutherodactylus coqui, Engystomops pustulosus, and Hymenochirus boettgeri. Frog chromosomes have remained remarkably stable since the Mesozoic Era, with limited Robertsonian (i.e., arm-preserving) translocations and end-to-end fusions found among the smaller chromosomes. Conservation of synteny includes conservation of centromere locations, marked by centromeric tandem repeats associated with Cenp-a binding surrounded by pericentromeric LINE/L1 elements. This work explores the structure of chromosomes across frogs, using a dense meiotic linkage map for X. tropicalis and chromatin conformation capture (Hi-C) data for all species. Abundant satellite repeats occupy the unusually long (~20 megabase) terminal regions of each chromosome that coincide with high rates of recombination. Both embryonic and differentiated cells show reproducible associations of centromeric chromatin and of telomeres, reflecting a Rabl-like configuration. Our comparative analyses reveal 13 conserved ancestral anuran chromosomes from which contemporary frog genomes were constructed.

For nearly four decades, harlequin toads, genus Atelopus, have suffered unparalleled population declines. While this also results in limited understanding of alphataxonomic relationships, these toads face an urgent need for advances in systematics to inform conservation efforts. However, high intraspecific variation and cryptic diversity have hindered a comprehensive understanding of Atelopus diversity. This is particularly exemplified among Amazonian populations related to A. spumarius, where decades of taxonomic work have not been able yet to unravel relationships between the many forms, while the names coined so far have led to taxonomic confusion leaving numerous lineages unnamed. A recent comprehensive phylogenetic study has revealed new insights into the systematics of harlequin toads with an emphasis on Amazonian forms, identifying several unnamed lineages. We here describe one of these evolutionary lineages as a new species, restricted to the Ecuadorian Amazon basin, in an integrative taxonomic approach using molecular, morphological, bioacoustic and larval information. With this, we contribute to a better understanding of Atelopus diversity as the baseline of conservation action. [ABSTRACT FROM AUTHOR]

Herein, basel on novel data gathered by citizens scientists and specialists, we contribute to the improvement of scientific knowledge and conservation of the Near Threatened Proceratophrys paviotii in order to: 1) test for the first time the phylogenetic position and a species delimitation of P. paviotii through a molecular approach; 2) describe a larger sample of its advertisement call to properly encompass the species intraspecific variation; 3) describe for the first time the P. paviotii release call; and 4) provide novel insights on the species conservation status. Our 16S tree confidently grouped P. paviotii with P. cururu, P. renalis, and P. laticeps. The average sequence divergence between P. paviotii and its congeners ranged from 2.2% (P. laticeps) to 9.1% (P. redacta). Advertisement calls consisted of a single note with duration of 0.26–0.58 s, 17–41 pulses emitted at rate of 54.19–77.49 pulses/s and peak frequency of 775.19–947.46 Hz. Release calls consisted of a single note with duration of 0.04–0.43 s, 2–13 pulses emitted at rate of 21.17–81.58 pulses/s and peak frequency of 689.1–1,722.6 Hz. Additionally, our study strongly supports the notion that Citizen Science approaches can yield invaluable information concerning species' geographic distribution and conservation. [ABSTRACT FROM AUTHOR]

The poison frog family (Anura: Dendrobatidae) consists of species with conspicuous (e.g., warning coloration and toxicity or low palatability) and cryptic (e.g., palatable, and inconspicuous coloration) traits. Previous literature suggests that conspicuous, but not cryptic, species require diet specialization in prey high in alkaloids. To test for dietary preferences of poison frog species, we identified, to the lowest possible taxonomic rank, the diets of 21 Epipedobates darwinwallacei (conspicuous) and 22 Hyloxalus awa (cryptic) frogs living in syntopy in the Otongachi Forest in northwestern Ecuador. We then tested for differences in diet assemblage composition, and diet specialization, in these putatively conspicuous and cryptic frogs. Our analyses showed significant differences in the composition of arthropod assemblages consumed by both frog species, which translated into a narrow niche breadth and nine arthropod taxa (out of a total of 18) consumed by both species. Moreover, the index of relative importance, which measures frog's diet specialization, suggested that E. darwinwallacei, and H. awa prefer specific arthropod taxa, where the former consumes preferentially springtails and mites, while the latter consumes mostly ants and Coleoptera larvae. Thus, contrary to expectations, diet specialization is not a unique characteristic of the species with conspicuous traits when living in syntopy. [ABSTRACT FROM AUTHOR]

Identifying and quantifying the main factors that are jeopardizing amphibian communities worldwide is essential for planning effective amphibian conservation. We collected data on the presence/absence of salamanders and newts (Caudata) and frogs and toads (Anura) in >3600 water points from a large region (30,000 km2) in NW Spain during a 10‐year period (2004–2013). We contrasted a large set of hypotheses explaining presence/absence as a function of anthropogenic factors across the regional elevation gradient (1–2036 m). Logistic regression modeling revealed that salamanders and newts were mainly influenced by pollution and by land use changes, and that the effect of these factors changed with elevation. However, frogs and toads were affected by a larger set of factors acting synergistically, although not including land use changes, and their effect operated at all elevation ranges, except in the case of biological factors (invasive species and wild boar abundance) with higher effects at low elevation. Changes in land use was the most common factor at any elevation and included the abandonment of rural areas, which favors shrub and tree encroachment on former open land, and loss of water points. The most resilient species at any elevation were two frogs, Pelophylax perezi and Rana parvipalmata. The least resilient species were two salamanders, Chioglossa lusitanica and Lissotriton helveticus, followed closely by two toads, Alytes obstetricans and Pelobates cultripes. Unoccupied sites had higher frequencies of biological effects and of changes in land use in the first 5 years of study and lower frequencies of direct human influence factors in the second period. Overall, our results showed that the studied amphibian metacommunity was negatively influenced both by direct and indirect anthropogenic factors, but also that many amphibian species were not only capable of occupying sites which had been altered by human action, but were even favored by land uses generating open habitat, a habitat type that is increasingly uncommon in the region, in the entire Iberian Peninsula and in Europe. [ABSTRACT FROM AUTHOR]

Tadpoles serve as crucial evidence for testing systematic and taxonomic hypotheses. Suctorial tadpoles collected in Guyana were initially assigned to Rhaebo nasicus through molecular phylogeny. Subsequent analysis of larval and adult morphological traits revealed synapomorphies within the clade encompassing R. nasicus and R. ceratophrys, prompting the recognition of a new genus described herein as Adhaerobufo. The new genus is distinguished from other bufonids by specific phenotypic traits including an enlarged, suctorial oral disc with distinct papillae arrangements, and the presence of certain muscles and narial vacuities at the larval stage. However, only a few adult external characteristics (e.g., enlarged eyelids, infraocular cream spot), seem to be reliably discriminative from related genera. This study underscores the significance of larval morphology in anuran systematics and offers new insights into the evolution of suctorial and gastromyzophorous larvae within bufonids. [ABSTRACT FROM AUTHOR]

Habitat alteration is recognized as one of the most important drivers of biodiversity decline. Urbanization implies a major habitat alteration and its impact on biodiversity depends on type and degrees of habitat modifications. Growing cities generate great heterogeneity with respect to the uses of space that enable the preservation of a certain diversity of amphibians within the urban frame. Our objective is to understand how environments with different levels of urbanization influence local amphibian diversity. We grouped various urban green spaces of a medium-sized and growing city of Argentina in relation to natural and anthropic environmental variables which allows us to describe an urban environmental gradient and we calculate beta diversity among these groups of urban environments considering the richness, composition and abundance of amphibian at each sites' group. From this, we analyze and tested the differences in the functional diversity of amphibians in the defined urban environmental gradient. We hypothesize that: (i) the described spatial heterogeneity would be related to an elevated beta diversity of amphibians within the urban frame; and (ii) the analysis of functional diversity would reveal a stronger biotic or abiotic filtering of the focal community. We found that groups of urban environments with an intermediate degree of urbanization can present a greater functional diversity and species richness than those that are less urbanized. We identified urban avoiders (T. typhonius, P. santafecinus and P. biligonigerus) and urban tolerant species (E. bicolor, R. arenarum, R. dorbignyi, L. macrosternum, L. latinasus, P. albonotatus, D. nanus and S. nasicus). The replacement beta diversity recorded indicated that the species assemblage changes throughout urban environmental gradient, but functionally similar species thrive in the less to the most urbanized sites, allowing them to occupy the entire functional space of the groups of urban environments with different types and degrees of urbanization. [ABSTRACT FROM AUTHOR]

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

Anurans exhibit a diverse acoustic repertoire, and the release call is emitted when an individual is improperly clasped, often by males mistakenly grasping other males or unreceptive females during amplexus. The release call also serves as a crucial source for taxonomic resolution. However, for certain widespread hylid genera like Trachycephalus, knowledge about their release calls is limited or non-existent. Trachycephalus comprises 18 species, yet only one has its release call described. This study presents the first description of the release call of T. typhonius from a population in the Cerrado of Central-Western Brazil. Its release call is composed of a multipulsed note, distinguishing it from the only other known call in the genus (T. 'vermiculatus'). Given the differences in the release call parameters between these species, such variations could serve as diagnostic criteria for distinguishing the two lineages. Additional descriptions of release calls, including from more T. typhonius and T. 'vermiculatus' populations, are essential for species diagnosis and taxonomic classification. [ABSTRACT FROM AUTHOR]

During their development, amphibians undergo various physiological processes that may affect their susceptibility to environmental pollutants. Naturally occurring fluctuations caused by developmental events are often overlooked in ecotoxicological studies. Our aim is to investigate how biomarkers of oxidative stress are modulated at different stages of larval development in the Amazonian amphibian species, Physalaemus ephippifer. The premetamorphosis, prometamorphosis and metamorphic climax stages were used to analyze total antioxidant capacity (ACAP), glutathione S-transferase (GST) activity, lipid peroxidation (LPO) levels and the expression of genes nrf2, gst, gsr (glutathione reductase) and gclc (glycine-cysteine ligase, catalytic subunit). Although there was no difference in ACAP and the genes expression among the studied stages, individuals from the premetamorphosis and prometamorphosis showed higher GST activity than ones under the climax. LPO levels were highest in individuals from the metamorphic climax. The present study suggests that the oxidative status changes during ontogeny of P. ephippifer tadpoles, especially during the metamorphic climax, the most demanding developmental phase. Variations in the redox balance at different developmental stages may lead to a divergent response to pollution. Therefore, we recommend that studies using anuran larvae as biomonitors consider possible physiological differences during ontogeny in their respective analyses. [ABSTRACT FROM AUTHOR]

We describe a new species of Centrolene from the Subandean Cordillera of Kutukú in southeastern Ecuador. The new species differs from all other glassfrogs by the combination of the following characters: presence of processes of vomers but without vomerine teeth; humeral spines in males; dorsum green with light green dots and without dark marks; dorsal skin with abundant tubercles; all visceral peritonea translucent (except for pericardium); and small body size (snout-vent length 21.5–21.9 mm in adult males). The new species is sister to Centrolene camposi from the Western Cordillera of the Andes of southwestern Ecuador, and together they form a clade with C. condor from the Subandean Cóndor Cordillera in southeastern Ecuador. Our time tree suggests that the new species originated at the end of the Pliocene. In addition, we present new information for C. zarza, expanding its geographic range across the southeastern Andes and the Kutukú and Cóndor cordilleras, amending its definition and diagnosis, and offering new information on its natural history and extinction risk. We also discuss the taxonomic status of Ecuadorian populations reported as C. huilensis and conclude that they are C. muelleri based on their close phylogenetic relationships and morphological similarity to samples of C. muelleri from Peru. Centrolene huilensis is a valid species and not closely related to C. muelleri. [ABSTRACT FROM AUTHOR]

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

Many anuran survival strategies involve hydric regulation, and reproduction is not different. The aquaporin (AQP) family plays an important role in water transport and regulation in many tissues, including the male gonad. The testes undergo various stages of change during the reproductive cycle, and water balance is an important factor for ensuring reproductive success. Considering the relevance of water control in testicular development in anurans and the lack of research regarding the tissue localization of AQP in the male gonad, the present study investigated the expression of three AQPs (1, 2, and 9) in the testis of the neotropical anuran species Leptodactylus podicipinus during two different periods of the reproductive cycle (reproductive and non-reproductive). AQP1 and 2 immunoreactions were found in early germ cells, spermatozoa, Leydig cells, and Sertoli cells, which were more frequently expressed within the reproductive period. AQP1 was also found in the testicular blood vessels. AQP9 was identified predominantly in the epithelium of the intratesticular ducts of reproductive-period individuals. This study presents, for the first time, the localization of AQP1, AQP2, and AQP9 in the testes of an anuran species and the differences in their location during two distinct periods of the reproductive cycle.

The Chinese horned toads, Boulenophrys boettgeri (Boulenger, 1899) and Boulenophrys kuatunensis (Pope, 1929), are two captivating species within the family Megophryidae, which inhabit the mountainous streams in the Eastern of China. In this study, two new complete mitochondrial genomes of B. boettgeri and B. kuatunensis were sequenced, assembled, and annotated using next-generation sequencing. The length of mitochondrial genomes of B. boettgeri and B. kuatunensis was 16,597 and 17,921 bp, respectively, with both containing 13 protein coding genes, 22 tRNA genes, two rRNA genes, and one putative control region. Phylogenetic relationships based on protein-coding mitochondrial genes showed that the two Boulenophrys species formed a cluster with other Boulenophrys species. The two new sequences provide valuable insights into the mitochondrial genomes of these two species, offering important data for understanding the phylogenetic relationships of the genus Boulenophrys.

This article provides a detailed description of the tadpole of the Hyloxalus nexipus frog species, focusing on its geographic variation and morphological characteristics. The researchers collected specimens from Ecuador and Peru and examined their coloration and measurements. The document emphasizes the importance of studying tadpoles in understanding anuran species. It also discusses the morphology and characteristics of tadpoles belonging to the Hyloxalus genus of poison dart frogs, noting specific differences in the upper jaw sheath and the position of the upper fin in Hyloxalus nexipus tadpoles. The information is based on detailed observations of single specimens in different developmental stages and references other relevant studies on tadpole morphology and taxonomy. [Extracted from the article]

Simple Summary: Head and neck cancers are aggressive and challenging to treat due to the severe side effects and toxicity of current treatments. Bufalin, a natural compound from the Chinese toad, has shown promise in fighting various cancers but has not been thoroughly studied for head and neck cancers. Our research aims to explore how bufalin works against these specific cancer cells. By using different techniques, we discovered that bufalin could reduce cancer cell growth, induce cell death, and enhance the body's immune response against tumors. These findings suggest that bufalin could become a new, effective treatment option with potentially fewer side effects for patients with head and neck cancers. This research could pave the way for developing better therapies and improving outcomes for patients facing this difficult disease. Bufalin, a cardiotonic steroid derived from the Chinese toad (Bufo gargarizans), has demonstrated potent anticancer properties across various cancer types, positioning it as a promising therapeutic candidate. However, comprehensive mechanistic studies specific to head and neck cancers have been lacking. Our study aimed to bridge this gap by investigating bufalin's mechanisms of action in head and neck cancer cells. Using several methods, such as Western blotting, immunofluorescence, and flow cytometry, we observed bufalin's dose-dependent reduction in cell viability, disruption of cell membrane integrity, and inhibition of colony formation in both HPV-positive and HPV-negative cell lines. Bufalin induces apoptosis through the modulation of apoptosis-related proteins, mitochondrial function, and reactive oxygen species production. It also arrests the cell cycle at the G2/M phase and attenuates cell migration while affecting epithelial–mesenchymal transition markers and targeting pivotal signaling pathways, including Wnt/β-catenin, EGFR, and NF-κB. Additionally, bufalin exerted immunomodulatory effects by polarizing macrophages toward the M1 phenotype, bolstering antitumor immune responses. These findings underscore bufalin's potential as a multifaceted therapeutic agent against head and neck cancers, targeting essential pathways involved in proliferation, apoptosis, cell cycle regulation, metastasis, and immune modulation. Further research is warranted to validate these mechanisms and optimize bufalin's clinical application. [ABSTRACT FROM AUTHOR]

In addition to the type locality (the summit of Aprada-tepui, Bolívar State of Venezuela), the distribution of the egg-brooding frog Stefania satelles was long thought to include several isolated tabletop mountain (tepui) summits surrounding the large Chimantá Massif in Bolívar State (hence the Latin name "satelles"). However, multilocus molecular phylogenetic analyses have revealed that this taxon includes several undescribed morphologically cryptic species, and that S. satelles should be restricted to its type locality. Two tepui-summit species confused under that name in the literature remain to be named, and the present paper aims at describing these populations previously referred to as Stefania sp. 3 and S. sp. 5. Stefania sp. 3 is only known from the small summit of Angasima-tepui, while S. sp. 5 is only reported from the small summit of Upuigma-tepui, both mountains being located south of the Chimantá Massif. These new, phylogenetically distinct species are described based on external morphology and osteology and in comparison to close relatives in the S. ginesi clade, which consists exclusively of tepui summit species. Both new species have highly restricted geographic ranges (less than 3 km2) and should be listed as Critically Endangered according to IUCN criteria. [ABSTRACT FROM AUTHOR]

Centrolenidae is a Neotropical family widely distributed in Central and South America, with its species richness concentrated in the tropical Andes. Several taxonomic problems have been identified within this family, mostly related to species with broad geographic distributions. In this study, we assessed and redefined the species boundaries of the Centrolene buckleyi species complex, and formally described two new species from the Andes of Ecuador. These new taxa are recognized by a combination of morphometric, osteological, acoustic, and genetic data. Following IUCN criteria, we propose that the two new species should to be considered as Endangered (EN), mainly because of their small distributions and habitat loss. The C. buckleyi complex provides insights into the biogeography of closely related Andean species. As in other glassfrogs, speciation in Centrolene seems to be mediated by the linearity of the Andes, where gene flow can be restricted by topography and, also, local extinctions. [ABSTRACT FROM AUTHOR]

The Chinese horned toads, Boulenophrys boettgeri (Boulenger, 1899) and Boulenophrys kuatunensis (Pope, 1929), are two captivating species within the family Megophryidae, which inhabit the mountainous streams in the Eastern of China. In this study, two new complete mitochondrial genomes of B. boettgeri and B. kuatunensis were sequenced, assembled, and annotated using next-generation sequencing. The length of mitochondrial genomes of B. boettgeri and B. kuatunensis was 16,597 and 17,921 bp, respectively, with both containing 13 protein coding genes, 22 tRNA genes, two rRNA genes, and one putative control region. Phylogenetic relationships based on protein-coding mitochondrial genes showed that the two Boulenophrys species formed a cluster with other Boulenophrys species. The two new sequences provide valuable insights into the mitochondrial genomes of these two species, offering important data for understanding the phylogenetic relationships of the genus Boulenophrys. [ABSTRACT FROM AUTHOR]

Background: Kurixalus lenquanensis Yu, Wang, Hou, Rao and Yang, 2017 was originally described, based on 14 adult male specimens from Lengquan Village, Mengzi, Yunnan, China. So far, this species is known only from south-eastern Yunnan and information on females of this species is not available. During the field surveys in 2023, two Kurixalus specimens (one female and one male) were collected from central eastern Yunnan (Shilin, Kunming, Yunnan, China). These two specimens were confirmed to be K. lenquanensis by molecular phylogenetic analyses, based on 16S rRNA sequences. New information: A female specimen of K. lenquanensis is described for the first time and morphological variation amongst populations of this species is provided. The diagnosis and distribution of this species are updated. [ABSTRACT FROM AUTHOR]

Photoreceptor oil droplets (ODs) are spherical organelles placed most commonly within the inner segment of the cone photoreceptors. Comprising neutral lipids, ODs can be either non-pigmented or pigmented and have been considered optically functional in various studies. Among living amphibians, ODs were only reported to occur in frogs and toads (Anura), while they are absent in salamanders and caecilians. Nonetheless, the limited understanding of their taxonomic distribution in anurans impedes a comprehensive assessment of their evolution and relationship with visual ecology. We studied the retinae of 134 anuran species, extending the knowledge of the distribution of ODs to 46 of the 58 currently recognized families, and providing a new perspective on this group that complements the available information from other vertebrates. The occurrence of ODs in anurans shows a strong phylogenetic signal, and our findings revealed that ODs evolved at least six times during the evolutionary history of the group, independently from other vertebrates. Although no evident correlation was found between OD occurrence, adult habits and diel activity, it is inferred that each independent origin involves distinct scenarios in the evolution of ODs concerning photic habits. Furthermore, our results revealed significant differences in the size of the ODs between nocturnal and arrhythmic anurans relative to the length of the cones' outer segment. [ABSTRACT FROM AUTHOR]

The distribution of the Coastal Bull Frog, Hoplobatrachus litoralis in Arunachal Pradesh was not certain for lack of any tangible report. The occurrence of this species in this Himalayan state of Arunachal Pradesh is reported herein based on recent collection of three individuals from a swampy area from the Senki Valley of the Itanagar Wildlife Sanctuary, Papum Pare district, Arunachal Pradesh. Brief diagnostic characters are also provided based on the observed individuals. [ABSTRACT FROM AUTHOR]

Urban environments expose wildlife to levels of anthropogenic noise they would not experience in rural areas (e.g., traffic noise), and research suggests that many species adjust their acoustic signals for optimal transmission in urban soundscapes. However, our understanding of anuran (order Anura) responses to noise pollution in urban environments of the southeastern United States is limited, particularly for species that can breed during winter. Our goal was to examine how vocal anuran advertisement call characteristics during winter varied with increasing distance from roadways in bottomland hardwoods of Louisiana, USA. We deployed acoustic recording units at two sites (i.e., rural and urban) perpendicular to Interstate 10 at 200‐, 400‐, and 600‐m intervals (i.e., close, middle, and far) from November 2019 to January 2020. We detected Cajun Chorus Frogs (Pseudacris fouquettei) and Cricket Frogs (Acris spp.) at our rural site, and only detected Cricket Frogs at our urban site. At the rural site, Cajun Chorus Frogs produced longer duration notes at the far location compared to the middle location. At the urban site, Cricket Frogs produced higher dominant frequency calls at the close location compared to the far and middle locations and longer duration notes at the far location compared to the close location. We were unable to account for additional factors in our models (e.g., temperature, noise levels), but our results generally align with previous research. Our study provides baseline data for future research to examine the potential effects of traffic noise on winter advertisement calls in locations with similar environmental conditions and species. [ABSTRACT FROM AUTHOR]

Understanding species composition across temporal and spatial scales through participatory monitoring has contributed to the development of several studies focused on biodiversity in Neotropical ecosystems. Habitat loss and the illegal collection of bromeliads pose significant threats to bromeligenous frogs, which depend on the rainwater collected between bromeliad leaves for egg and tadpole development. In this study, we compiled a comprehensive dataset of bromeligenous frogs using data from "Projeto Bromélias" on the iNaturalist citizen science platform. Our dataset includes records of 85 species of bromeligenous frogs, representing 52% of the 164 known species that reproduce in bromeliads. These species belong to 33 genera and 10 families and are reported from 18 countries. Twenty-eight species are listed in threatened categories on a global scale. Our findings extended the known geographic distribution of four species. Notably, the green and black poison dart Frog, Dendrobates auratus, was recorded as a non-native species in Hawaii, USA. Regarding the temporal data, the number of bromeligenous records increased substantially after 2009. This study highlights the value of citizen science platforms as important tools for monitoring bromeliad inhabitants and contributing to management and conservation initiatives. [ABSTRACT FROM AUTHOR]

A new species of the genus Boulenophrys is described from the coastal hills of eastern Fujian Province, China. The new taxon can be distinguished from all recognized congeners by a combination of discrete morphological character state differences and genetic divergences in the combined mitochondrial 16S + CO1 genes. We also provide a map showing the distribution pattern of Boulenophrys species in Fujian and a provincial-specific key, which will aid their conservation by helping the local authorities accurately identify species during field identifications and data collection efforts. [ABSTRACT FROM AUTHOR]

Animals constantly need to evaluate available external and internal information to make appropriate decisions. Identifying, assessing, and acting on relevant cues in contexts such as mate choice, intra-sexual competition, and parental care is particularly important for optimizing individual reproductive success. Several factors can influence decision-making, such as external environmental cues and the animal's own internal state, yet, we have limited knowledge on how animals integrate available information. Here, we used an entire island population (57 males, 53 females, and 1,109 tadpoles) of the neotropical brilliant-thighed poison frog Allobates femoralis to investigate how 2 factors (olfactory cues and personality traits) influence the ability of males to find and use new resources for tadpole deposition. We experimentally manipulated the location of tadpole deposition sites and their associated olfactory cues, and repeatedly measured exploration and boldness in adult males. We further reconstructed tadpole deposition choices via inferred parent–offspring relationships of adult frogs and tadpoles deposited in our experimental pools using molecular parentage analysis. We found that the discovery and use of new rearing sites were heavily influenced by olfactory cues; however, we did not find an effect of the measured behavioral traits on resource discovery and use. We conclude that in highly dynamic environments such as tropical rainforests, reliable external cues likely take priority over personality traits, helping individuals to discover and make use of reproductive resources. [ABSTRACT FROM AUTHOR]

Advertisement calls are the main communication form of anurans, and other individuals can use it to evaluate several aspects of the calling individual. In this context, environmental disturbances, such as traffic noise, can potentially affect this recognition. Therefore, this study aims to evaluate the response of Aplastodiscus leucopygius to traffic noise in a fragment of Atlantic Forest within the city of São Paulo. The experimentation consisted of recording the calling individual previously, during and after an exposure to urban noise. After that, individuals were measured to evaluate the Scaled Mass Index (SMI), and individual and environmental temperatures were taken. Also, considering that individuals of this species present harmonic shifting, we tried to evaluate which factors (individual, acoustic, or environmental) are associated with this phenomenon. We observed that the individuals showed an increase in call activity after exposure to traffic noise, but none of the evaluated aspects here could explain the harmonic shifting in their calls. Considering that this increasing on call activity also means an increasing of individual's expense of energy, traffic noise is potentially harmful to the communication of A. leucopygius and, consequently, to its permanence in the site. [ABSTRACT FROM AUTHOR]

Anthropogenic activity and urbanization subject the diversity and distribution of amphibians to constant change. This study aims to reveal the impact of increasing urbanization pressure on amphibian diversity, density, and distribution in Istanbul, a megacity. I researched amphibian species' presence and dispersal in and around the wetlands of Istanbul, spanning urban, semi-urban, and non-urban areas. I conducted surveys in 2016 and 2017 at 396 sites and found seven species (four families) of amphibians at 151 stations. Anurans occurred at 38.13% of sites, and newts at 8.6%. The alpha diversity of semi-urban regions was higher than in urban and non-urban areas of Istanbul. Nonmetric multidimensional scaling (NMDS) analysis of relative abundances of amphibian species at the three site types showed significant differences. Compared to the distribution of each species reported in the literature, I found that distributions have spread to the west, east, and north of the province. Several species occurred in semi-urban wetlands due to the pressure of urbanization, which causes extirpations or declines to low abundance, making their detection unlikely in urban areas. [ABSTRACT FROM AUTHOR]

Nonvisual opsins are transmembrane proteins expressed in the eyes and other tissues of many animals. When paired with a light-sensitive chromophore, nonvisual opsins form photopigments involved in various nonvisual, light-detection functions including circadian rhythm regulation, light-seeking behaviors, and seasonal responses. Here, we investigate the molecular evolution of nonvisual opsin genes in anuran amphibians (frogs and toads). We test several evolutionary hypotheses including the predicted loss of nonvisual opsins due to nocturnal ancestry and potential functional differences in nonvisual opsins resulting from environmental light variation across diverse anuran ecologies. Using whole-eye transcriptomes of 81 species, combined with genomes, multitissue transcriptomes, and independently annotated genes from an additional 21 species, we identify which nonvisual opsins are present in anuran genomes and those that are also expressed in the eyes, compare selective constraint among genes, and test for potential adaptive evolution by comparing selection between discrete ecological classes. At the genomic level, we recovered all 18 ancestral vertebrate nonvisual opsins, indicating that anurans demonstrate the lowest documented amount of opsin gene loss among ancestrally nocturnal tetrapods. We consistently found expression of 14 nonvisual opsins in anuran eyes and detected positive selection in a subset of these genes. We also found shifts in selective constraint acting on nonvisual opsins in frogs with differing activity periods, habitats, distributions, life histories, and pupil shapes, which may reflect functional adaptation. Although many nonvisual opsins remain poorly understood, these findings provide insight into the diversity and evolution of these genes across anurans, filling an important gap in our understanding of vertebrate opsins and setting the stage for future research on their functional evolution across taxa. [ABSTRACT FROM AUTHOR]

The repeated evolution of gliding in diverse Asian vertebrate lineages is hypothesized to have been triggered by the dominance of tall dipterocarp trees in the tropical forests of Southeast Asia. These dipterocarp forests have acted as both centres of diversification and climatic refugia for gliding vertebrates, and support most of their extant diversity. We predict similarities in the diversification patterns of dipterocarp trees and gliding vertebrates, and specifically test whether episodic diversification events such as rate shifts and/or mass extinctions were temporally congruent in these groups. We analysed diversification patterns in reconstructed timetrees of Asian dipterocarps, the most speciose gliding vertebrates from different classes (Draco lizards, gliding frogs and Pteromyini squirrels) and compared them with similar-sized clades of non-gliding relatives (Diploderma lizards, Philautus frogs and Callosciurinae squirrels) from Southeast Asia. We found significant declines in net-diversification rates of dipterocarps and the gliding vertebrates during the Pliocene-Pleistocene, but not in the non-gliding groups. We conclude that the homogeneity and temporal coincidence of these rate declines point to a viable ecological correlation between dipterocarps and the gliding vertebrates. Further, we suggest that while the diversification decay in dipterocarps was precipitated by post-Miocene aridification of Asia, the crises in the gliding vertebrates were induced by both events concomitantly.

Amphibians exhibit an incredible diversity of reproductive and life-history strategies, including various forms of nest construction and nesting behaviour. Although anuran amphibians (frogs and toads) are not known for their nests, nesting behaviour in this clade-broadly defined as a location chosen or constructed for eggs and young-is tightly linked to the amphibious lifestyle of this group. Transitions to increasingly terrestrial living have driven reproductive diversity in anurans, including the repeated, independent evolution of nests and nesting. Indeed, a core feature of many notable anuran adaptations-including nesting behaviour-is the maintenance of an aquatic environment for developing offspring. The tight link between increasingly terrestrial reproduction and morphological, physiological and behavioural diversity in anurans provides inroads for studying the evolutionary ecology of nests, their architects and their contents. This review provides an overview of nests and nesting behaviour in anurans, highlighting areas where additional work may be particularly fruitful. I take an intentionally broad view of what constitutes nesting to highlight what we can learn from thinking and researching comparatively across anurans and vertebrates more broadly. This article is part of the theme issue The evolutionary ecology of nests: a cross-taxon approach.

The phylogeny of the major lineages of Amphibia has received significant attention in recent years, although evolutionary relationships within families remain largely neglected. One such overlooked group is the subfamily Holoadeninae, comprising 73 species across nine genera and characterized by a disjunct geographical distribution. The lack of a fossil record for this subfamily hampers the formulation of a comprehensive evolutionary hypothesis for their diversification. Aiming to fill this gap, we inferred the phylogenetic relationships and divergence times for Holoadeninae using molecular data and calibration information derived from the fossil record of Neobatrachia. Our inferred phylogeny confirmed most genus-level associations, and molecular dating analysis placed the origin of Holoadeninae in the Eocene, with subsequent splits also occurring during this period. The climatic and geological events that occurred during the Oligocene-Miocene transition were crucial to the dynamic biogeographical history of the subfamily. However, the wide highest posterior density intervals in our divergence time estimates are primarily attributed to the absence of Holoadeninae fossil information and, secondarily, to the limited number of sampled nucleotide sites.

The identification of conservation priority areas for amphibians in Iran remains unexplored. Although protected areas play a vital role in biodiversity conservation, 40 percent of amphibians are at risk of extinction. The present study aimed to identify the priority areas for amphibian conservation and assess the effectiveness of the current protected conservation areas. 2192 species occurrence points for 23 amphibians, including 16 species of Anura and 7 species of Caudata, were curated. These occurrence points in addition to bioclimatic, anthropogenic, and geographical variables were used to model the species distribution. Then, the richness and rarity maps of amphibians were determined and hotspots identified based on the top 10 % area of the country with the highest richness and rarity values. Finally, Iran's protected areas were overlaid to evaluate the current coverage of hotspots and identify future conservation priorities. Although approximately 12 % of Iran is currently protected, the present study findings indicate that the current network of protected areas is ineffective for amphibian conservation; over 90 % of hotspots for amphibians lie outside the current national protected area network. The most important hotspots are located in the Caspian Hyrcanian Mixed Forest, the western margin of the North Zagros, and the Central Zagros Mountains Forest-Steppe ecoregions of the country. Among different types of protected areas that overlap with amphibian hotspots, protected areas and wildlife refuges respectively ranked the highest in quantity and size, while national parks ranked the lowest. In the current study, a baseline of top candidate areas for expanding protected areas where habitats can be managed to protect amphibians in Iran was provided. To further improve protected areas' coverage, priority conservation areas in the Zagros Mountains Forest-Steppe hotspots were recommended.

Habitat alteration is recognized as one of the most important drivers of biodiversity decline. Urbanization implies a major habitat alteration and its impact on biodiversity depends on type and degrees of habitat modifications. Growing cities generate great heterogeneity with respect to the uses of space that enable the preservation of a certain diversity of amphibians within the urban frame. Our objective is to understand how environments with different levels of urbanization influence local amphibian diversity. We grouped various urban green spaces of a medium-sized and growing city of Argentina in relation to natural and anthropic environmental variables which allows us to describe an urban environmental gradient and we calculate beta diversity among these groups of urban environments considering the richness, composition and abundance of amphibian at each sites’ group. From this, we analyze and tested the differences in the functional diversity of amphibians in the defined urban environmental gradient. We hypothesize that: (i) the described spatial heterogeneity would be related to an elevated beta diversity of amphibians within the urban frame; and (ii) the analysis of functional diversity would reveal a stronger biotic or abiotic filtering of the focal community. We found that groups of urban environments with an intermediate degree of urbanization can present a greater functional diversity and species richness than those that are less urbanized. We identified urban avoiders (T. typhonius, P. santafecinus and P. biligonigerus) and urban tolerant species (E. bicolor, R. arenarum, R. dorbignyi, L. macrosternum, L. latinasus, P. albonotatus, D. nanus and S. nasicus). The replacement beta diversity recorded indicated that the species assemblage changes throughout urban environmental gradient, but functionally similar species thrive in the less to the most urbanized sites, allowing them to occupy the entire functional space of the groups of urban environments with different types and degrees of urbanization.

The emergence of complex social interactions is predicted to be an important selective force in the diversification of communication systems. Parental care presents a key social context in which to study the evolution of novel signals, as care often requires communication and behavioral coordination between parents and is an evolutionary stepping-stone toward increasingly complex social systems. Anuran amphibians (frogs and toads) are a classic model of acoustic communication and the vocal repertoires of many species have been characterized in the contexts of advertisement, courtship, and aggression, yet quantitative descriptions of calls elicited in the context of parental care are lacking. The biparental poison frog, Ranitomeya imitator, exhibits a remarkable parenting behavior in which females, cued by the calls of their male partners, feed tadpoles unfertilized eggs. Here, we characterized and compared calls across three social contexts, for the first time including a parental care context. We found that egg-feeding calls share some properties with both advertisement and courtship calls but also had unique properties. Multivariate analysis revealed high classification success for advertisement and courtship calls but misclassified nearly half of egg feeding calls as either advertisement or courtship calls. Egg feeding and courtship calls both contained less identity information than advertisement calls, as expected for signals used in close-range communication where uncertainty about identity is low and additional signal modalities may be used. Taken together, egg-feeding calls likely borrowed and recombined elements of both ancestral call types to solicit a novel, context-dependent parenting response.

Genome and cell size are strongly correlated across species1,2,3,4,5,6 and influence physiological traits like developmental rate.7,8,9,10,11,12 Although size scaling features such as the nuclear-cytoplasmic (N/C) ratio are precisely maintained in adult tissues,13 it is unclear when during embryonic development size scaling relationships are established. Frogs of the genus Xenopus provide a model to investigate this question, since 29 extant Xenopus species vary in ploidy from 2 to 12 copies (N) of the ancestral frog genome, ranging from 20 to 108 chromosomes.14,15 The most widely studied species, X. laevis (4N = 36) and X. tropicalis (2N = 20), scale at all levels, from body size to cellular and subcellular levels.16 Paradoxically, the rare, critically endangered dodecaploid (12N = 108) Xenopus longipes (X. longipes) is a small frog.15,17 We observed that despite some morphological differences, X. longipes and X. laevis embryogenesis occurred with similar timing, with genome to cell size scaling emerging at the swimming tadpole stage. Across the three species, cell size was determined primarily by egg size, whereas nuclear size correlated with genome size during embryogenesis, resulting in different N/C ratios in blastulae prior to gastrulation. At the subcellular level, nuclear size correlated more strongly with genome size, whereas mitotic spindle size scaled with cell size. Our cross-species study indicates that scaling of cell size to ploidy is not due to abrupt changes in cell division timing, that different size scaling regimes occur during embryogenesis, and that the developmental program of Xenopus is remarkably consistent across a wide range of genome and egg sizes.

The global emergency of antimicrobial resistance has drawn several efforts to evaluate new drug candidates, such as natural defensive biomolecules. Ocellatins are a group of antimicrobial peptides found in anurans of the Leptodactylidae family. This work investigated the presence of antimicrobial peptides in the skin secretion of Leptodactylus vastus from the Brazilian northeast. The secretion was fractionated by RP-HPLC, and the fractions were screened for antibacterial activity. A peptide isolated from the most active fraction was characterized for primary structure and evaluated for antibacterial activity, cytotoxicity to murine melanoma cells (B16-F10), and hemolytic activity. The RP-HPLC profile displayed 26 fractions, with fraction 25 being the most active. One of the two peptides present in this fraction had the primary structure determined, belonging to the group of ocellatins. Since it was not identical to other ocellatins previously reported, it was named ocellatin-VT. This peptide especially inhibited Gram-negative bacteria growth, with the highest activity against Acinetobacter baumannii and Escherichia coli (growth inhibition was higher than 95% at 8 and 16 µM, respectively). Ocellatin-VT was weakly cytotoxic to B16-F10 cells and showed low hemolytic activity. In conclusion, a new ocellatin was isolated from L. vastus skin secretion that was active against non-resistant and multidrug-resistant bacteria.

Aposematism continues to be a phenomenon of central interest in evolutionary biology. The life history of the mimic poison frog, Ranitomeya imitator, relies heavily on aposematism. In order for aposematic signals to be effective, predators must be able to learn to avoid the associated phenotype. However, in R. imitator, aposematism is associated with four different color phenotypes that mimic a complex of congeneric species occurring across the mimic frog's geographic range. Investigations of the underlying mechanics of color production in these frogs can provide insights into how and why these different morphs evolved. We used histological samples to examine divergence in the color production mechanisms used by R. imitator to produce effective aposematic signals across its geographic range. We measured the coverage of melanophores and xanthophores (the area covered by chromatophores divided by total area of the skin section) in each color morph. We find that morphs that produce orange skin exhibit a higher coverage of xanthophores and lower coverage of melanophores than those that produce yellow skin. In turn, morphs that produce yellow skin exhibit a higher coverage of xanthophores and lower coverage of melanophores than those that produce green skin. Generally, across the morphs, a high ratio of xanthophores to melanophores is associated with colors of brighter spectral reflectance. Together, our results contribute to the understanding of color production in amphibians and document divergence in the histology of a species that is subject to divergent selection associated with aposematism.

Herein, we describe a new species of terrestrially-nesting fanged frog from Sulawesi Island, Indonesia. Though male nest attendance and terrestrial egg deposition is known in one other Sulawesi fanged frog (Limnonectes arathooni), the new species exhibits a derived reproductive mode unique to the Sulawesi assemblage; male frogs guard one or more clutches of eggs festooned to leaves or mossy boulders one to two meters above small slow-moving streams, trickles, or seeps. This island endemic has thus far been collected at three sites on Sulawesi: one in the Central Core of the island, and two on the Southwest Peninsula-south of the Tempe Depression (a major biogeographical boundary). The new Limnonectes has the smallest adult body size among its Sulawesi congeners-with a maximum snout-vent length of about 30 millimeters. Beyond its unique reproductive behavior and body size, the species is further diagnosed on the basis of advertisement call and genetic distance from sympatric fanged frogs. The discovery and description of the new species highlights the remarkable reproductive trait diversity that characterizes the Sulawesi fanged frog assemblage despite that most species in this radiation have yet to be formally described.

True poison-dart frogs (Phyllobates, Dendrobatidae) evolved the ability to secrete batrachotoxins, the most powerful alkaloids known to date. The genus comprises five species whose systematics, at first glance, appeared clear. The most derived clade would include two Colombian species (P. terribilis and P. bicolor) with the highest toxicity, the largest body size, and predominantly yellow body colouration. The other three species (P. aurotaenia, P. vittatus, and P. lugubris) are less toxic on average, have smaller size, and are predominantly black with bright dorsolateral stripes. Recent research has revealed the existence of two major lineages among the three Colombian species. The northern lineage appears to result from a complex evolutionary history, including perhaps introgression among yellow and black taxa. The southern lineage instead revealed the existence of new clades closely related to P. terribilis, black and yellow, that arguably deserve their recognition as new species. Here, available evidence is combined to support the erection of southern populations of P. aurotaenia as a new highly toxic species, sister to P. terribilis, and much closer to it than to any other yellow or black-bodied species, Phyllobates samperi sp. nov. Their common ancestor is sister to an additional yellow species, which we also describe here as Phyllobates bezosi sp. nov. Both new species can be externally diagnosed using colouration. Our previous and current analyses also suggest the existence of additional taxa and corroborate multiple transitions in colouration across these hypertoxic taxa. [ABSTRACT FROM AUTHOR]

The behavioural responses of predator-naïve (laboratory-born), predator-experienced (laboratory-born but with short-term experience with predators), and wild-caught (with long-term experience with predators in natural settings) tadpoles of Sphaerotheca breviceps to carnivorous predatory tadpoles of Hoplobatrachus tigerinus was studied in the laboratory. The predator's stimulus solution (kairomones) was used to simulate predation threat. All S. breviceps tadpoles (i.e., predatornaïve, predator-experienced and wild-caught) exhibited behavioural antipredator responses, i.e., reduced swimming activity and time spent swimming, and had a higher burst speed in response to water-borne kairomonal cues of predators. Wildcaught tadpoles showed stronger responses than predator-naïve and predator-experienced tadpoles and the latter exhibited stronger responses than predator-naïve tadpoles. Our study thus suggests that antipredator behaviour in these tadpoles is basically innate. Furthermore, the enhanced antipredator behaviour of wild-caught tadpoles indicates that learning is involved. [ABSTRACT FROM AUTHOR]

The Alliance of Genome Resources (Alliance) is an extensible coalition of knowledgebases focused on the genetics and genomics of intensively studied model organisms. The Alliance is organized as individual knowledge centers with strong connections to their research communities and a centralized software infrastructure, discussed here. Model organisms currently represented in the Alliance are budding yeast, Caenorhabditis elegans , Drosophila , zebrafish, frog, laboratory mouse, laboratory rat, and the Gene Ontology Consortium. The project is in a rapid development phase to harmonize knowledge, store it, analyze it, and present it to the community through a web portal, direct downloads, and application programming interfaces (APIs). Here, we focus on developments over the last 2 years. Specifically, we added and enhanced tools for browsing the genome (JBrowse), downloading sequences, mining complex data (AllianceMine), visualizing pathways, full-text searching of the literature (Textpresso), and sequence similarity searching (SequenceServer). We enhanced existing interactive data tables and added an interactive table of paralogs to complement our representation of orthology. To support individual model organism communities, we implemented species-specific "landing pages" and will add disease-specific portals soon; in addition, we support a common community forum implemented in Discourse software. We describe our progress toward a central persistent database to support curation, the data modeling that underpins harmonization, and progress toward a state-of-the-art literature curation system with integrated artificial intelligence and machine learning (AI/ML). [ABSTRACT FROM AUTHOR]

Taxonomy and species boundaries within the Rhacophorus rhodopus and Rhacophorus bipunctatus complexes are very confusing. In this study, we attempt to delimit the species boundaries and test the currently accepted taxonomic assignments within these two complexes based on newly collected samples and previously published data across their distributions. Phylogenetic analyses revealed that the R. rhodopus and R. bipunctatus complexes consisted of six distinct clades (labeled A‒F) that diverged from each other by genetic distances (p-distance) ranging from 5.3% to 9.2% in 16S rRNA sequences, and accordingly analyses of species delimitation placed them into six species, of which three correspond to known species (R. rhodopus, R. bipunctatus, and R. napoensis) and three represent different cryptic species. Rhacophorus rhodopus (Clade C) is distributed in southern Yunnan, China, northern Laos, and northern and central Thailand; R. bipunctatus (Clade F) is distributed in northeastern India and western and northern Myanmar; and R. napoensis (Clade B) is distributed in Guangxi, China and northern Vietnam. Based on both molecular and morphological evidence, we described the clade consisting of samples from Hainan, China and central Vietnam (Clade A) as a new species, Rhacophorus qiongica sp. nov. There are two cryptic species requiring additional morphological studies: one only contains samples from Motuo, Xizang, China (Clade E), and the other is distributed in western Yunnan, China, central Myanmar, central Thailand, and Malaysia (Clade D). Additionally, our results supported the idea that some old GenBank sequences of R. reinwardtii need to be updated with the correct species name. [ABSTRACT FROM AUTHOR]

We describe a new frog species that is the sibling of Pristimantis reichlei. These two sister species inhabit the Amazonian lowlands and adjacent foothills of the Andes, from central Bolivia to central Peru. Pristimantis reichlei occurs from central Bolivia to southern Peru (Alto Purús National Park), while the new species occurs from northern Bolivia (Departamento Pando) to Panguana in central Peru (Departamento Huánuco), at elevations between 220 and 470 m a.s.l. In spite of their morphological crypsis, these siblings occur in syntopy without evidence of interbreeding (in the Alto Purús area) and are recovered as reciprocally monophyletic. Their uncorrected pairwise genetic distances in the 16S rRNA gene range from 9.5–13.5%, and their advertisement calls differ in both qualitative and quantitative traits. Moreover, our study found uncorrected pairwise distances within the new species of up to 5.0% and up to 9.3% within P. reichlei. We therefore cannot rule out the possible existence of hybrids or additional species-level lineages hidden in this complex. Furthermore, we found another potential pair of sibling species composed of P. danae and an unnamed lineage, with divergences of 9.4% in the 16S gene, whose in-depth analysis and taxonomic treatment are pending future revision. With the new nominal species, the Pristimantis danae species group now includes 20 species, distributed across the upper Amazon basin and in the eastern Andes, from western Brazil to Bolivia and Peru. Our study, together with an increasing number of other studies, indicates that sibling species are far from being rare among Amazonian amphibians and that species resolution remains low even for groups that have received considerable attention in recent years. [ABSTRACT FROM AUTHOR]

Recent phylogenetic studies in Himalayan lazy toads of the genus Scutiger Theobald, 1868 revealed the presence of genetically deeply divergent lineages. The taxonomy of S. nepalensis sensu lato was re-assessed based on museum material considering molecular and morphological data. The results strongly support the recognition of a new species, S. kanjiroba sp. nov. distributed along the Nepalese Kanjiroba massif. It is further shown that S. sikimmensis has an apparently much more restricted distribution range than previously thought. The frequent misidentification of Scutiger across large areas of the Himalaya-Tibet area highlights the significance of careful taxonomic evaluation of collection material and the need for the direct morphological comparison of closely related species when describing new species. [ABSTRACT FROM AUTHOR]

We report the first mitochondrial genomes of the Afrobatrachian frog species Hyperolius substriatus, providing insights into the gene arrangements, duplications, and evolutionary dynamics within this taxonomically complex genus. Assemblies were gained from two museum specimens and included all 37 typical vertebrate mitogenomic genes. In both cases, the mitogenome of H. substriatus shows duplications of tRNAs (trnL, trnT and trnP). The WANCY region exhibits the characteristic Afrobatrachian pattern but with unique intergenic spacers. Both copies of duplicated non-coding and control regions show high sequence similarity within and between specimens. The distinct gene order and homology of duplications observed can be explained under the tandem duplication and random loss model of mitochondrial evolution. Furthermore, a comparison with Hyperolius marmoratus highlights differences in gene copy number and synteny, and the presence of a duplication of trnM in the latter appears to be a derived condition. Together, these findings provide valuable insights into the taxonomy and evolution of Hyperolius, contributing to understanding interspecific gene reorganisation in this diverse group and suggesting that gene arrangements could help resolve some of the taxonomic problems of this complex genus. [ABSTRACT FROM AUTHOR]