Larval morphology and natural history in two Minervarya species from Andaman Islands, with comments on a new phytotelmonous larval morphotype for the genus.

We studied the larval morphology and natural history of two species of Minervarya from the Andaman Islands of India. A recent assignment of Charles Darwins frog to the genus Minervarya opened up new questions on the systematic affinities and natural history of its enigmatic tadpoles. Originally described as Rana charlesdarwini Das, 1998, and long considered as Ingerana charlesdarwini, the tadpoles of this species were poorly understood and incomparable to those of other presumed close relatives based on an original brief description of its phytotelmonous tadpoles. Here we present a detailed redescription of the larval morphology of M. charlesdarwini based on genetically confirmed tadpoles. At the same time, we compared the external morphology of M. charlesdarwini tadpoles to that of its closest phylogenetic and sympatric congener, M. andamanensis, consequently reporting the first-ever larval description for M. andamanensis. For the first time also for minervaryan frogs, we record that M. charlesdarwini lays eggs on the inner walls of tree holes, and the tadpoles possess morphological adaptations for a phytotelmonous life. Their unique features, such as tail-to-body ratio >2.2, reduced labial tooth rows (LTRF 1/1), acutely pointed serrations in the jaw sheath, and the medial portion of the upper sheath broadly arched and that of the lower sheath U-shaped, justify adding a new larval morphotype to the genus Minervarya. In contrast, M. andamanensis breeds and undergoes development in ground water puddles and ponds, a common trait for other known minervaryan species. Furthermore, our study generates new knowledge on the amplexus, oviposition site, and early embryonic development in these species. The deeper understanding of the biology and habitat requirements contributes additional traits for future systematic and evolutionary studies, and helps guide conservation assessments for these two species endemic to a handful of islands in the Andaman Archipelago.

Hiding in Plain Sight: Rain Water Puddles in Nicobar Islands of India Reveal Abundance of a New Frog Species of the Genus Microhyla Tschudi, 1838 (Anura: Microhylidae).

Recent studies on frogs of the Microhyla heymonsi species complex have demonstrated that high genetic variation exists among its various known populations from regions across Asia. We assessed the taxonomic identity of the Nicobar population of Microhyla cf. heymonsi and compared it to the typical Microhyla heymonsi from Taiwan and the two recently described species in this complex from Vietnam. Our study demonstrates that the Nicobar population is both genetically and morphologically divergent and warrants recognition as a new species, which we formally describe here as Microhyla nakkavaram sp. nov. The new species is closely related to M. daklakensis, M. heymonsi, and M. ninhthuanensis, but diagnosable from all three species by a suite of morphological characters, such as the presence of two small tubercles at mid-dorsum along with ( )-shaped markings, length of finger I longer than half the length of finger II, presence of three distinct metacarpal tubercles on hand, rudimentary foot webbing, as well as its slender body shape, granular dorsal skin texture, and other colour characters and body markings. Statistical analyses based on multiple morphometric characters also clearly separate our new taxon from M. heymonsi, with which it was previously confused. Our phylogenetic analyses based on the mitochondrial 16S rRNA locus find Microhyla nakkavaram sp. nov. to be nested in the Microhyla achatina species group, where it is delimited as a distinct species. This lineage shows genetic distances of ≥ 3.5% from all the other known congeners. Currently, the known distribution of the new taxon is restricted to the southernmost group of Nicobar Islands-Great Nicobar, Kondul, and Little Nicobar-where it is found in abundance across a wide range of habitats during the monsoon season. Our study underlines the need to reassess the identity of all the known populations of M. 'heymonsi' from other regions in Southeast and East Asia. To facilitate future taxonomic work in the light of our and other recent findings, we also provide a detailed redescription and revised diagnosis for M. heymonsi based on morphological examination of its century-old type material originating from Taiwan.

A tiny new species of Microhyla Tschudi, 1838 (Amphibia: Anura: Microhylidae) from Belitung Island and Southeastern Sumatra, Indonesia.

A tiny new species of narrow-mouthed frog of the genus Microhyla is described from the island of Belitung and southeastern Sumatra, Indonesia. The most distinctive feature of the new frog is its diminutive adult size, snout-vent length ranging only from 12.3 to 15.8 mm in adult males. Phylogenetic analyses based a fragment of the mitochondrial 16S rRNA gene, along with detailed morphological and acoustic comparison differentiate the new taxon from all known congeners. The new species, formally described as Microhyla sriwijaya sp. nov., is a member of the M. achatina species group and the sister taxon to M. orientalis. It is diagnosable from other congeners by a combination of characters: (1) smaller male snout-vent size 16 mm; (2) snout obtusely rounded in dorsal view; (3) absence of mid-dorsal line and skin fold; (4) first finger reduced (finger I length less than half of finger II length); (5) dorsum with a prominent dark median mark extending posteriorly, narrow anteriorly near the level of the shoulder and expanding dorsolaterally up to the vent; margins of the dorsal marking concave with broad reddish-brown or orange colouration on either side; (6) foot webbing rudimentary, reaching just up to the first subarticular tubercle on all toes; (7) dorsal skin with prominent tubercles, especially in life; (8) tibiotarsal articulation of adpressed limb reaching beyond the snout tip; and (9) males produce a single type of call with pulsatile temporal structure, calls of relatively short duration ranging between 31.862.8 s, with two to three pulses delivered at a rate ranging between 32.236.0 pulses per second, and the mean overall dominant frequency of 4.3 kHz. The uncorrected pairwise genetic distances between Microhyla sriwijaya sp. nov. and all other known congeners are > 3.8% for the studied 16S gene fragment. The new species was discovered from wayside rural areas with oil palm plantations at four localities in the small island of Belitung (type locality), and from coffee plantation and secondary forest at Lampung in southeastern Sumatra. It is not known from any protected area and appears to be threatened due to tin mining activity, intensive logging, oil palm, and other commonly practiced agriculture activities.

An integrative approach to infer systematic relationships and define species groups in the shrub frog genus Raorchestes, with description of five new species from the Western Ghats, India.

The genus Raorchestes is a large radiation of Old World tree frogs for which the Western Ghats in Peninsular India is the major center for origin and diversification. Extensive studies on this group during the past two decades have resolved long-standing taxonomic confusions and uncovered several new species, resulting in a four-fold increase in the number of known Raorchestes frogs from this region. Our ongoing research has revealed another five new species in the genus, formally described as Raorchestes drutaahu sp. nov., Raorchestes kakkayamensis sp. nov., Raorchestes keirasabinae sp. nov., Raorchestes sanjappai sp. nov., and Raorchestes vellikkannan sp. nov., all from the State of Kerala in southern Western Ghats. Based on new collections, we also provide insights on the taxonomic identity of three previously known taxa. Furthermore, since attempts for an up-to-date comprehensive study of this taxonomically challenging genus using multiple integrative taxonomic approaches have been lacking, here we review the systematic affinities of all known Raorchestes species and define 16 species groups based on evidence from multi-gene (2,327 bp) phylogenetic analyses, several morphological characters (including eye colouration and pattern), and acoustic parameters (temporal and spectral properties, as well as calling height). The results of our study present novel insights to facilitate a better working taxonomy for this rather speciose and morphologically conserved radiation of shrub frogs. This will further enable proper field identification, provide momentum for multi-disciplinary studies, as well as assist conservation of one of the most colourful and acoustically diverse frog groups of the Western Ghats biodiversity hotspot.

New insights on the systematics and reproductive behaviour in tree frogs of the genus Feihyla, with description of a new related genus from Asia (Anura, Rhacophoridae).

The taxonomy of the Asian tree frog genus Feihyla has been in a state of flux ever since its proposal in 2006. Allocation of species to Feihyla remains confusing, particularly with respect to the closely related genus Chirixalus (formerly Chiromantis sensu lato). At the same time, several Chirixalus species are known only from cursory descriptions and remain poorly studied. In this study, we review the systematics of the genus Feihyla and clarify the generic placement of its members along with all other species currently assigned to Chirixalus. Based on integrative evidence gathered from new collections, examination of types and original descriptions, morphological comparisons, phylogenetic relationships inferred from a multi-gene (three mitochondrial + two nuclear) 1,937 bp dataset, as well as reproductive modes including egg-laying, nesting behaviour, and clutch morphology, our results show that the six species formerly attributed to Feihyla represent three morphologically and phylogenetically distinct groups: (1) the type species of the genus, F. palpebralis, along with F. fuhua constitute the Feihyla palpebralis group or Feihyla sensu stricto; (2) F. inexpectata and F. kajau represent the Feihyla vittiger group and are closely related to the Feihyla palpebralis group; (3) 'F. hansenae' and 'F. vittata' constitute another distinct and reciprocally monophyletic lineage, more closely related to Chirixalus + Chiromantis. In light of long-standing taxonomic confusions and unresolved phylogenetic relationships, we propose recognition of a new genus to accommodate 'F. hansenae' and 'F. vittata' in order to stabilise the classification of several Asian rhacophorid species that have been frequently confused and transferred within the Chirixalus-Chiromantis-Feihyla complex. Based on integrative evidence, eight species previously attributed to the genus Chirixalus or Feihyla are formally transferred to Rohanixalus gen. nov., two Chirixalus members are allocated to Feihyla, and one synonymised with Rhacophorus bipunctatus. In addition, we report the first member of the tree frog family Rhacophoridae from the Andaman Islands of India-Rohanixalus vittatus, along with description of its male advertisement call, reproductive behaviour including parental care by the female, and larval morphology. Extended distributions are also provided for Rohanixalus species across Northeast India. The study further reveals the presence of potentially undescribed diversity in the new genus. Altogether, the revised classification and novel insights presented herein will facilitate a better working taxonomy for four phylogenetically distinct but morphologically related groups of Old World tree frogs.

A new Microhyla species (Anura: Microhylidae) from riparian evergreen forest in the eastern Himalayan state of Arunachal Pradesh, India.

A new frog species of the genus Microhyla (Anura, Microhylidae) is described from riparian mid-elevation (860 m asl) evergreen forest in Namdapha National Park, located in the eastern Himalayan state of Arunachal Pradesh, India. The new species can be morphologically distinguished from other congeners by a suite of characters such as adult size, dorsal and lateral colouration and markings, snout shape, foot webbing, and digit tip morphology. Phylogenetically, the new species is more closely related to some of the smallest known members of the genus. It forms a deeply divergent sister lineage to the clade containing members of the Microhyla zeylanica species group that are restricted to Peninsular India and Sri Lanka, and shows sequential relationship with Southeast Asian species M. superciliaris, followed by clade containing M. aurantiventris + M. butleri. The discovery indicates that novel taxa representing distinct evolutionary lineages still remain to be formally described in the genus Microhyla, especially from less explored regions such as the eastern Himalayan forests in Northeast India.

Rediscovery of Micryletta inornata (Boulenger, 1890) from Sumatra: redescription, molecular identity, and taxonomic implications.

Micryletta inornata (Boulenger 1890), the type species of the genus Micryletta, was originally described from the island of Sumatra in Indonesia. Subsequently, this species has been widely reported from Sundaland (Sumatra and Malay Peninsula), Indo-China, Northeast India and South Andaman, up to southern China and Taiwan. However, since the original description there has been no further report of this species from the type locality or the island. During a herpetofaunal survey in Sumatra, several specimens that are morphologically concordant with the original description and the syntypes of M. inornata were found, and thus the species was rediscovered after 125 years. Here, we provide a redescription of the species based on the freshly collected specimens, along with a detailed morphological and molecular comparison with known congeners. Further, using molecular data from the mitochondrial 16S rRNA gene, our study recovered the Sumatran M. inornata as a phylogenetically distinct lineage from all other populations previously referred to this species. This confirms that all known Micryletta 'inornata' populations from regions outside Sumatra constitute several other lineages representing either new species or previously available names currently considered as synonyms, consequently requiring taxonomic validation in the future.

Author Correction: New microhylid frog genus from Peninsular India with Southeast Asian affinity suggests multiple Cenozoic biotic exchanges between India and Eurasia.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Multiple Independent Recruitment of Sodefrin Precursor-Like Factors in Anuran Sexually Dimorphic Glands.

Chemical signaling in animals often plays a central role in eliciting a variety of responses during reproductive interactions between males and females. One of the best-known vertebrate courtship pheromone systems is sodefrin precursor-like factors (SPFs), a family of two-domain three-finger proteins with a female-receptivity enhancing function, currently only known from salamanders. The oldest divergence between active components in a single salamander species dates back to the Late Paleozoic, indicating that these proteins potentially gained a pheromone function earlier in amphibian evolution. Here, we combined whole transcriptome sequencing, proteomics, histology, and molecular phylogenetics in a comparative approach to investigate SPF occurrence in male breeding glands across the evolutionary tree of anurans (frogs and toads). Our study shows that multiple families of both terrestrially and aquatically reproducing frogs have substantially increased expression levels of SPFs in male breeding glands. This suggests that multiple anuran lineages make use of SPFs to complement acoustic and visual sexual signaling during courtship. Comparative analyses show that anurans independently recruited these proteins each time the gland location on the male's body allowed efficient transmission of the secretion to the female's nares.

A new species of Micryletta frog (Microhylidae) from Northeast India.

We describe a new species of frog in the microhylid genus Micryletta Dubois, 1987 from Northeast India based on molecular and morphological evidence. The new species, formally described as Micryletta aishani sp. nov., is phenotypically distinct from other congeners by a suite of morphological characters such as brown to reddish-brown dorsum; dorsal skin shagreened with minute spinules; snout shape nearly truncate in dorsal and ventral view; a prominent dark streak extending from tip of the snout up to the lower abdomen; ash-grey mottling along the margins of upper and lower lip extending up to the flanks, limb margins and dorsal surfaces of hand and foot; tibiotarsal articulation reaching up to the level of armpits; absence of outer metatarsal tubercles; and absence of webbing between toes. Phylogenetic relationships within the genus are inferred based on mitochondrial data and the new taxon is found to differ from all the recognised Micryletta species by 3.5-5.9% divergence in the mitochondrial 16S rRNA. The new species was found in the states of Assam, Manipur, and Tripura, from low to moderate elevation (30-800 m asl) regions lying south of River Brahmaputra and encompassing the Indo-Burma Biodiversity Hotspot. The discovery validates the presence of genus Micryletta in Northeast India based on genetic evidence, consequently confirming the extension of its geographical range, westwards from Southeast Asia up to Northeast India. Further, for nomenclatural stability of two previously known species, Microhyla inornata (= Micryletta inornata) and Microhyla steinegeri (= Micryletta steinegeri), lectotypes are designated along with detailed descriptions.

New microhylid frog genus from Peninsular India with Southeast Asian affinity suggests multiple Cenozoic biotic exchanges between India and Eurasia.

Anurans in Peninsular India exhibit close biogeographical links with Gondwana as well as Laurasia, often explainable by the geological history of the Indian subcontinent; its breakup from Gondwanan landmasses followed by long isolation that resulted in diversification of endemic lineages, and subsequent land connections with Asia that enabled dispersal of widespread groups. Although widely distributed, the frog subfamily Microhylinae mostly comprises of geographically restricted genera found either in Southeast and East Asia or Peninsular India and Sri Lanka. Here we report a previously unknown microhylid from the Western Ghats in Peninsular India with closest relatives found over 2,000 km away in Southeast Asia. Based on integrated evidence from mitochondrial and nuclear DNA, adult and tadpole morphology, hand musculature, male advertisement call, and geographical distance, we recognize this enigmatic frog as a distinct new species and genus endemic to the Western Ghats. The discovery of Mysticellus franki gen. et sp. nov. and its close evolutionary relationship with the Southeast Asian genus Micryletta also provide insights on the biogeography of Microhylinae. Genus-level divergences within the subfamily suggest multiple Cenozoic biotic exchange events between India and Eurasia, particularly through postulated Eocene land bridges via Southeast Asia prior to accretion of the two landmasses.

Cryptic diversity within the Megophrys major species group (Amphibia: Megophryidae) of the Asian Horned Frogs: Phylogenetic perspectives and a taxonomic revision of South Asian taxa, with descriptions of four new species.

The Megophrys major species group (MMSG) is composed of typically medium to large sized frogs. Within the genus, it is the most geographically widespread clade ranging from the western Himalayas to southern Indochina. In this study, we examined in detail the extent of cryptic diversity within the MMSG-Indian populations based on molecular data (up to ten genes) using multigene concatenation and coalescent-based phylogenetic techniques, species delimitation analyses and extensive morphological data.Molecular evidence suggests a high level of hidden cryptic diversity within the MMSG, particularly within the M. major species complex (MMC), highlighting overlapping distributions, a case of potential mitochondrial transfer between two species, and tree topology discordance between phylogenetic methods and mitochondrial and nuclear data sets. Most analyses indicated distinct eastern and western clades in the MMC, and that the western clade may further divide into a northern and a southern subclade.A detailed taxonomic review of Indian members of the Megophrys major species group is provided. Previously undocumented complex nomenclatural issues involving known species are highlighted and resolved. Megophrys monticola is taxonomically redefined for the first time as a valid species since its synonymy with M. parva in 1893. The taxonomic status of two recently described species, Xenophrys katabhako and X. sanu are discussed in light of increased molecular and morphological sampling, and are synonymised with M. monticola. Megophrys monticola and M. robusta are redescribed based on their original type specimens and recently collected material. Megophrys major is neotypified and M. robusta lectotypified to remove prevailing nomenclatural instability. Four new large sized Indian MMC species are formally described from the Northeast Indian states of Arunachal Pradesh, Meghalaya and Nagaland, and Myanmar. All South Asian MMSG species are morphologically diagnosed from known congeners in the group. The geographic distributions of all taxa discussed are significantly redefined based on the revised taxonomy and extensive literature review. Morphological and molecular evidence suggests that Megophrys major sensu stricto might be endemic to Northeast India; M. glandulosa is formally removed from the Indian and Bhutan species checklists. Numerous misidentifications in literature are highlighted and corrected. In order to reduce future misidentifications of species reported from surrounding regions, high definition images of the holotypes of three Chinese species, M. glandulosa, M. medogensis and M. zhangi are provided for the first time, and a detailed description of Myanmar specimens of M. glandulosa is also given. This study provides the principle foundation for further research into the taxonomic status of the remaining, currently undescribed MMC taxa from Southeast Asia.

Delineating Microhyla ornata (Anura, Microhylidae): mitochondrial DNA barcodes resolve century-old taxonomic misidentification.

Microhyla ornata, a species originally described from the southwest coast of India in 1841, was long reported to be wide-ranging throughout South, Southeast, and East Asia. Although the name M. ornata is restricted to populations from South Asia, the species is still considered to occur widely in India and its neighboring regions. To clarify the identity and geographical distribution of M. 'ornata', we performed DNA barcoding using a fragment of the mitochondrial 16S rRNA gene from 62 newly obtained samples. Our results show that this taxon is restricted to Peninsular India and Sri Lanka, whereas, populations from the other parts represent three different species - M. mukhlesuri, M. mymensinghensis, and M. nilphamariensis, creating new country records for India. Our work reemphasizes the benefits of DNA barcoding for rapidly identifying populations of widespread species and provides insights into the patterns of genetic differentiation in the M. 'ornata' species complex of South Asia.

Seven new species of Night Frogs (Anura, Nyctibatrachidae) from the Western Ghats Biodiversity Hotspot of India, with remarkably high diversity of diminutive forms.

The Night Frog genus Nyctibatrachus (Family Nyctibatrachidae) represents an endemic anuran lineage of the Western Ghats Biodiversity Hotspot, India. Until now, it included 28 recognised species, of which more than half were described recently over the last five years. Our amphibian explorations have further revealed the presence of undescribed species of Nights Frogs in the southern Western Ghats. Based on integrated molecular, morphological and bioacoustic evidence, seven new species are formally described here as Nyctibatrachus athirappillyensis sp. nov., Nyctibatrachus manalari sp. nov., Nyctibatrachus pulivijayani sp. nov., Nyctibatrachus radcliffei sp. nov., Nyctibatrachus robinmoorei sp. nov., Nyctibatrachus sabarimalai sp. nov. and Nyctibatrachus webilla sp. nov., thereby bringing the total number of valid Nyctibatrachus species to 35 and increasing the former diversity estimates by a quarter. Detailed morphological descriptions, comparisons with other members of the genus, natural history notes, and genetic relationships inferred from phylogenetic analyses of a mitochondrial dataset are presented for all the new species. Additionally, characteristics of male advertisement calls are described for four new and three previously known species. Among the new species, six are currently known to be geographically restricted to low and mid elevation regions south of Palghat gap in the states of Kerala and Tamil Nadu, and one is probably endemic to high-elevation mountain streams slightly northward of the gap in Tamil Nadu. Interestingly, four new species are also among the smallest known Indian frogs. Hence, our discovery of several new species, particularly of easily overlooked miniaturized forms, reiterates that the known amphibian diversity of the Western Ghats of India still remains underestimated.

Evolutionary History of the Asian Horned Frogs (Megophryinae): Integrative Approaches to Timetree Dating in the Absence of a Fossil Record.

Molecular dating studies typically need fossils to calibrate the analyses. Unfortunately, the fossil record is extremely poor or presently nonexistent for many species groups, rendering such dating analysis difficult. One such group is the Asian horned frogs (Megophryinae). Sampling all generic nomina, we combined a novel ∼5 kb dataset composed of four nuclear and three mitochondrial gene fragments to produce a robust phylogeny, with an extensive external morphological study to produce a working taxonomy for the group. Expanding the molecular dataset to include out-groups of fossil-represented ancestral anuran families, we compared the priorless RelTime dating method with the widely used prior-based Bayesian timetree method, MCMCtree, utilizing a novel combination of fossil priors for anuran phylogenetic dating. The phylogeny was then subjected to ancestral phylogeographic analyses, and dating estimates were compared with likely biogeographic vicariant events. Phylogenetic analyses demonstrated that previously proposed systematic hypotheses were incorrect due to the paraphyly of genera. Molecular phylogenetic, morphological, and timetree results support the recognition of Megophryinae as a single genus, Megophrys, with a subgenus level classification. Timetree results using RelTime better corresponded with the known fossil record for the out-group anuran tree. For the priorless in-group, it also outperformed MCMCtree when node date estimates were compared with likely influential historical biogeographic events, providing novel insights into the evolutionary history of this pan-Asian anuran group. Given a relatively small molecular dataset, and limited prior knowledge, this study demonstrates that the computationally rapid RelTime dating tool may outperform more popular and complex prior reliant timetree methodologies.

Description of four new species of Burrowing Frogs in the Fejervarya rufescens complex (Dicroglossidae) with notes on morphological affinities of Fejervarya species in the Western Ghats.

The Rufescent Burrowing Frog, Fejervarya rufescens, is thought to have a wide distribution across the Western Ghats in Peninsular India. This locally abundant but secretive species has a short breeding period, making it a challenging subject for field studies. We sampled 16 populations of frogs morphologically similar to F. rufescens in order to understand the variation among populations found across the Western Ghats. Our study shows significant morphological and genetic differences among the sampled populations, suggesting that F. 'rufescens' is a complex of several undescribed species. Using evidence from morphology and genetics, we confirm the presence of five distinct species in this group and formally describe four as new. The new species were delineated using a phylogeny based on three mitochondrial genes (16S, COI and Cytb) and a haplotype network of a nuclear gene (Rag1). Hereafter, the distribution of F. rufescens is restricted to the state of Karnataka and adjoining regions of northern Kerala. Three new species (Fejervarya kadar sp. nov., Fejervarya manoharani sp. nov. and Fejervarya neilcoxi sp. nov.) are from regions south of Palghat gap in the state of Kerala, and one (Fejervarya cepfi sp. nov.) from the northern Western Ghats state of Maharashtra. These findings indicate that Fejervarya frogs of the Western Ghats are more diverse than currently known. Our results will also have implications on the conservation status of F. rufescens, which was previously categorized as Least Concern based on its presumed wide geographical distribution. Furthermore, in order to facilitate a better taxonomic understanding of this region's fejervaryan frogs, we divide all the known Fejarvarya species of the Western Ghats into four major groups-Fejervarya nilagirica group, Fejervarya rufescens group, Fejervarya sahyadris group and Fejervarya syhadrensis group, based on their morphological affinities.

Molecular and Morphological Study of Leaping Frogs (Anura, Ranixalidae) with Description of Two New Species.

The monotypic anuran family Ranixalidae is endemic to India, with a predominant distribution in the Western Ghats, a region that is home to several unique amphibian lineages. It is also one of the three ancient anuran families that diversified on the Indian landmass long before several larger radiations of extant frogs in this region. In recent years, ranixalids have been subjected to DNA barcoding and systematic studies. Nearly half of the presently recognized species in this family have been described over the last three years, along with recognition of a new genus to accommodate three previously known members. Our surveys in the Western Ghats further suggest the presence of undescribed diversity in this group, thereby increasing former diversity estimates. Based on rapid genetic identification using a mitochondrial gene, followed by phylogenetic analyses with an additional nuclear gene and detailed morphological studies including examination of museum specimens, new collections, and available literature, here we describe two new species belonging to the genus Indirana from the Western Ghats states of Karnataka and Kerala. We also provide new genetic and morphological data along with confirmed distribution records for all the species known prior to this study. This updated systematic revision of family Ranixalidae will facilitate future studies and provide vital information for conservation assessment of these relic frogs.

A new species of Microhyla (Anura: Microhylidae) from Sri Lanka: an integrative taxonomic approach.

Species boundaries of Microhyla rubra of India and Sri Lanka were assessed using the following criteria: genetic barcoding, morphology, and vocalization. We use a ca. 500 bp fragment of the 16S rRNA mitochondrial gene and show that there is an uncorrected pairwise distance of 2.7-3.2% between the Indian and Sri Lankan populations of M. rubra. We show that they are different in several call characteristics such as, dominant frequency, call duration, call rise time and pulse rate. Morphologically, the Sri Lankan population can be distinguished from the typical M. rubra described from southern India, by a combination of characters: body size, skin texture, and feet dimensions. We recognize the population from Sri Lanka as a new species, Microhyla mihintalei sp. nov., a widely distributed lowland species with an elevational distribution of up to 500 m a.s.l.

On the taxonomic status of Ichthyophis husaini Pillai & Ravichandran, 1999 (Amphibia: Gymnophiona: Ichthyophiidae).

Extensive surveys at and around the type locality of Ichthyophis husaini Pillai & Ravichandran, 1999 (an 'unstriped' Ichthyophis) in North East India between 2008 and 2014 yielded tens of striped but no unstriped specimens of Ichthyophis. Contrary to the original description, we observed a lateral stripe on the holotype of I. husaini. The holotype of I. husaini is very similar to the holotype of the striped species I. garoensis Pillai & Ravichandran, 1999, which was collected from a locality only 20 km away from the type locality of I. husaini. The holotypes of these two species are also very similar to 12 newly collected specimens of striped Ichthyophis from this area. We conclude that the holotypes of I. husaini and I. garoensis represent but a single species, and we designate Ichthyophis husaini Pillai & Ravichandran, 1999 as a junior subjective synonym of Ichthyophis garoensis Pillai & Ravichandran, 1999. This taxonomic action and newly reported material extends the known geographic range of I. garoensis. It also reduces the number of Ichthyophis species of North East India to eight, and restricts unstriped members of the genus within this region to a single species (I. sikkimensis Taylor, 1960) that is known only from the far west of the region.