Complete mitochondrial genome of the boreal digging frog Kaloula borealis (Anura, Microhylidae)

We sequenced the complete mitochondrial genome from the boreal digging frog Kaloula borealis. The genome sequence was 17,173 bp in size, and the gene order and contents were identical to those of previously reported amphibian mitochondrial genomes. Of 13 protein-coding genes (PCGs), 5 genes (CO2, ATPase 6, CO3, ND3, and ND4) had incomplete stop codons. Also ND1 gene used GTG as a start codon, while CO1 and ND5 genes used AGG as a stop codon. The base composition of K. borealis mitogenome showed a strong anti-G bias (6.11%) on the 3rd position of PCGs.     

Morphology of the spermatozoa of the Microhylidae (Anura, Amphibia)

Microhylid spermatozoa show the autapomorphic condition of possessing a thin post-mitochondrial cytoplasmic collar. Their spermatozoa are apomorphic in several respects. They have lost the distinct nuclear shoulder, endonuclear canal and axial perforatorium observed in urodeles, caecilians and primitive frogs, possess a conical perforatorium and apomorphically lack any fibres associated with the axoneme. The spermatozoa of Cophixalus , however, differ in several respects from those of the other microhylids examined. Cophixalus spermatozoa are longer in almost all measurements, the acrosome vesicle is cylindrical and does not completely cover the putative perforatorium, the perforatorium is asymmetrical and composed of fine fibres, the nucleus is strongly attenuated and narrower, and the mitochondria are elongate. The absence of fibres associated with the axoneme is an apomorphic condition shared with the Ranidae, Rhacophoridae and Pipidae.     

At the lower size limit for tetrapods, two new species of the miniaturized frog genus Paedophryne (Anura, Microhylidae)

I describe two new species in the miniaturized microhylid frog genus Paedophryne from forests in southeastern Papua New Guinea. The first species is described on the basis of two specimens and exhibits female snout-vent length of 8.5-9.0 mm (no males known), whereas that of the second species, described on the basis of 12 specimens, is 8.8-9.3 mm, with males 8.1-8.9 mm. These frogs are smaller than the other two diminutive species described when the genus was recently erected, and they represent what are currently the smallest known species of tetrapods. The two species replace each other elevationally on the same mountain massif and occur in relative geographic proximity to the other named species of the genus. Females of both species contain only two enlarged ova, suggesting that they also possess clutch sizes at the extreme lower end of variation in frogs. All species of Paedophryne inhabit leaf litter, as seen for most other miniaturized anurans.     

Burrowing behavior of Dermatonotus muelleri (Anura,Microhylidae) with reference to the origin of the burrowing behavior of Anura

Dermatonotus muelleri is a forelimbs-head-first burrowing frog that uses its forelimbs for soil removal, and it is the second anuran species known to arch its head downwards at an angle of almost 90° to the longitudinal axis of its body when burrowing. The burrowing behavior of D. muelleri is divided in three stages: head burrowing, body burrowing, and chamber construction. Burrowing in D. muelleri includes construction of a subterranean chamber used for estivation during the dry season. Phylogenetic analysis based on literature survey of burrowing behavior suggested that head-first burrowing behavior has evolved several times in anuran history, forming a convergence complex, and that hindlimbs-first burrowing is a basal behavior.     

Molecular systematics of the Middle American genus Hypopachus (Anura: Microhylidae)

We present the first phylogenetic study on the widespread Middle American microhylid frog genus Hypopachus. Partial sequences of mitochondrial (12S and 16S ribosomal RNA) and nuclear (rhodopsin) genes (1275 bp total) were analyzed from 43 samples of Hypopachus, three currently recognized species of Gastrophryne, and seven arthroleptid, brevicipitid and microhylid outgroup taxa. Maximum parsimony (PAUP), maximum likelihood (RAxML) and Bayesian inference (MrBayes) optimality criteria were used for phylogenetic analyses, and BEAST was used to estimate divergence dates of major clades. Population-level analyses were conducted with the programs NETWORK and Arlequin. Results confirm the placement of Hypopachus and Gastrophryne as sister taxa, but the latter genus was strongly supported as paraphyletic. The African phrynomerine genus Phrynomantis was recovered as the sister taxon to a monophyletic Chiasmocleis, rendering our well-supported clade of gastrophrynines paraphyletic. Hypopachus barberi was supported as a disjunctly distributed highland species, and we recovered a basal split in lowland populations of Hypopachus variolosus from the Pacific versant of Mexico and elsewhere in the Mesoamerican lowlands. Dating analyses from BEAST estimate speciation within the genus Hypopachus occurred in the late Miocene/early Pliocene for most clades. Previous studies have not found bioacoustic or morphological differences among these lowland clades, and our molecular data support the continued recognition of two species in the genus Hypopachus.     

Three new species of Oreophryne (Anura,Microhylidae) from Papua New Guinea

I describe three new species of the diverse microhylid frog genus Oreophryne from Papua New Guinea. Two of these occur in two isolated mountain ranges along the northern coast of Papua New Guinea; the third is from Rossel Island in the very southeasternmost part of the country. All three are the first Oreophryne known from these areas to have a cartilaginous connection between the procoracoid and scapula, a feature usually seen in species far to the west or from the central cordillera of New Guinea. Each of the new species also differs from the many other Papuan Oreophryne in a variety of other morphological, color-pattern, and call features. Advertisement-call data for Oreophryne species from the north-coast region suggest that they represent only two of the several call types seen in regions further south, consistent with the relatively recent derivation of these northern regions as accreted island-arc systems. The distinctively different, whinnying, call type of the new species from Rossel Island occurs among other Oreophryne from southeastern Papua New Guinea but has been unreported elsewhere, raising the possibility that it may characterize a clade endemic to that region.     

A Complete Embryonic Developmental Table of Microhyla fissipes(Amphibia,Anura, Microhylidae)

Access to embryonic developmental stages is essential basic work for understanding how organisms develop. In this study, seven egg clutches(range 209–564 eggs) of ornamented pygmy frog Microhyla fissipes(Amphibia, Anura, Microhylidae) were obtained from seven breeding pairs in laboratory. One egg clutch of them was observed for the embryonic development, and the staging table of normal development was constructed based on morphological and physiological characteristics. Forty-five developmental stages were defined for M. fissipes, and two major developmental periods were designated: 1) early embryonic development period(stages 1–28), from fertilization to operculum completion stage, lasted for 82.6 hours at water temperature(WT) 23–25℃; 2) larval development period(stages 29–45), from operculum completion to tail complete absorption stage, took 38 days at WT 22–26.5℃, showing that the embryos of this species develop rapidly. In addition, the tadpoles were transparent, which is similar to those in field. These characteristics suggest that M. fissipes would be a good model to study developmental biology, adaptive mechanisms from aquatic to terrestrial phases, environmental toxicology, and human disease.     

A New Species of Microhyla (Anura: Microhylidae) from Nilphamari,Bangladesh

A new species of Microhyla frog from the Nilphamari district of Bangladesh is described and compared with its morphologically similar and geographically proximate congeners. Molecular phylogeny derived from mitochondrial DNA sequences revealed that although the new species – designated here as Microhyla nilphamariensis sp. nov. – forms a clade with M. ornate, it is highly divergent from M. ornata and all of its congeners, with 5.7 – 13.2% sequence divergence at the 16S rRNA gene. The new species can be identified phenotypically on the basis of a set of diagnostic (both qualitative and quantitative) characters as follows: head length is 77% of head width, distance from front of eyes to the nostril is roughly six times greater than nostril–snout length, internarial distance is roughly five times greater than nostril–snout length, interorbital distance is two times greater than internarial distance, and distance from back of mandible to back of the eye is 15% of head length. Furthermore, inner metacarpal tubercle is small and ovoid-shaped, whereas outer metacarpal tubercle is very small and rounded. Toes have rudimentary webbing, digital discs are absent, inner metatarsal tubercle is small and round, outer metatarsal tubercle is ovoid-shaped, minute, and indistinct.     

Chromosomal analysis of twelve species of Microhylidae (Anura) from Madagascar

The karyotypes of four species of Dyscophinae and eight species of Cophylinae were analyzed. The chromosome number was 2n=26 in all cases. Between the two subfamilies a difference in the form of the karyotype was observed; the chromosomes show a gradual decrease in length in the Dyscophinae, whereas in the Cophylinae the karyotype demonstrates a clear discontinuity of size between pairs 5 and 6.Chromosomal polymorphism was found in Plethodontohyla tuberata, the chromosomes of pair 4 were subtelocentric in the homozygous specimens, whereas this pair showed a subtelocentric and a submetacentric chromosome of equal length in the heterozygous one, suggesting a pericentric inversion. Although in the Cophylinae the chromosome number is constant, the number of chromosome arms is variable. Pericentric inversions seem to play an important role in the chromosomal evolution of the Cophylinae.     

饰纹姬蛙求偶鸣声特征分析

2012年5月,用SX950录音笔和Praat声音分析软件对浙江丽水繁殖季节饰纹姬蛙(Microhyla ornata)求偶鸣声进行录制和特征分析.结果表明,饰纹姬蛙发出的求偶鸣声具有单一谐波鸣声结构、多脉冲(7、9～16)及纺锤形振幅等特征;所有鸣声主频率范围为1.22～4.09 kHz (n=233),且由不同脉冲数组成的鸣声主频率平均值几近相等;叫声时程随脉冲数的增加而增大,脉冲时程在不同脉冲数鸣声中的大小几近相等,但最后一个脉冲的时程大小≤其他脉冲;脉冲间隔与叫声时程则刚好相反,即叫声时程越短,脉冲间隔就越大.在7个脉冲的鸣声中,其脉冲间隔最大,脉冲率最小;而在16个脉冲的鸣声中,脉冲间隔则最小,脉冲率最大.除7个脉冲和16个脉冲鸣声的脉冲率分别与其他鸣声存在显著性差异以外,随着叫声时程和脉冲数的增加,脉冲率也出现相应变化.在声强方面,除16个脉冲鸣声与其余所有的脉冲鸣声出现显著性差异以外,其他脉冲鸣声之间的两两比较差异不显著.丽水种群与其他5个地理种群(杭州、宣城、Kamoor、Bajipe和Padil)的鸣声特征比较显示饰纹姬蛙在不同地理种群的鸣声结构相似,而鸣声主频率、叫声时程、脉冲时程及脉冲率等在6个地理种群种均出现不同程度的差异.了解不同物种的声信号特征有助于更好地理解动物通讯行为及其进化特点.     

A New Record of Kaloula(Amphibia: Anura: Microhylidae) in Shanghai,China

We discovered a medium-sized species of narrow-mouthed frog of the genus Kaloula in June and September 2014 during municipal surveys of amphibians in Shanghai. Three narrow-mouthed frogs were collected in city drains of the Binjiang Forest Park, Shanghai. Based on subsequent examination and morphological characters, these specimens were identified as Kaloula borealis(Barbour, 1908). The species was mainly distributed in Northern China and Korea. It is a new amphibian record for Shanghai. We found the visible differences in male secondary sexual characteristics between our specimens and Beijing specimens. K. borealis was first described inhabiting urban greenlands which could benefit amphibian conservation in urban and urbanizing areas.     

The orientation of muscle fibres in the tail musculature ofRana temporaria tadpoles (Amphibia,Anura)

Summary Shape of the myosepts and arrangement of the muscle fibres were recorded in the lateral musculature of the tail ofRana temporaria embryos and larvae. Well developed myomeres are present as early as st. 18–19. The main characteristics—ie. those related to functional properties—of myoseptal shape as well as of muscle fibre arrangement, remain unchanged throughout further development until degeneration of the tail occurs during metamorphosis. The rather simple myoseptal shape observed inRana—as compared to the multiple cone-form observed in most fishes—shows a close agreement to hypothetical myosept models described in papers by Jarman (1961), van der Stelt (1968) and Willemse (1966). The muscle fibres in the m. lateralis ofRana are arranged in trajectorial patterns that show a close similarity to the trajectorial patterns observed in typical teleosts. Both arrangements agree with trajectorial models based on the mathematical analyses of Alexander (1968).Neurulas anaesthetized with 1:10000 MS-222 and exposed up two weeks to this anaesthetic developed the same shape of the myosepts and arrangement of muscle fibres as in controls. Thus even the details of the function-related features of the myomere structure develop without functioning. In this field possible feedback meachisms are either not affected by anaesthesia or do not exist at all.     

The submandibular musculature of phyllomedusinae (Anura: Hylidae): A reappraisal

The submandibular musculature of 37 species of the five currently recognized genera of the subfamily Phyllomedusinae (Anura: Hylidae) is described; observations are made on the variation and ontogeny of these muscles. Supplementary apical elements of the m. intermandibularis occur in all phyllomedusines studied, in addition to the supplementary posterolateral elements previously reported. Our observations are discussed in the context of 1) the proposed homology between supplementary apical and posterolateral elements; 2) the homology with the apical elements reported for Pelodryadinae (sister taxon of Phyllomedusinae); and 3) the implications for our understanding of the relationships between Phyllomedusinae and Pelodryadinae. Anatomical differences between the apical and posterolateral elements and their co‐occurrence in phyllomedusines indicate that these supplementary elements are not homologous. Despite differences between phyllomedusines and pelodryadines in the adhesion of supplementary fibers to the principal element of the m. intermandibularis and the occurrence of a broad aponeurosis or a medial raphe, the extensive morphological and developmental resemblances of the apical elements indicate that these structures are homologous, and that the presence of apical elements is a synapomorphy of Phyllomedusinae + Pelodryadinae. J. Morphol., 2011. © 2011 Wiley‐Liss, Inc.     

A New Species of Genus Microhyla(Amphibia: Anura: Microhylidae) from Zhejiang Province,China

We described a new species, Microhyla beilunensis sp. nov., from Zhejiang Province of China. Phylogenetic analyses based on the mitochondrial 12 S, 16 S and CO1 gene sequences suggested that the new taxon was distinctly separated from its congeners and closed to M. mixtura and M. okinavensis. Morphologically, the new species could be identified from its congeners except M. mixtura by several characters:(1) rudimentary webs on toe base;(2) absence of disks and dorsal median longitudinal grooves on finger tips;(3) presence of disks and dorsal median longitudinal grooves on toe tips. As well, the new species could be identified from topotype M. mixtura by the combination of characters:(1) apart from the stripes, bar-shaped and oval-shaped patterns, the rounded spots present on the dorsum of body and legs;(2) the outer metacarpal tubercles prominently larger than the inner one;(3) of males, the ratios of HW, IND, UEW and LAW to SVL of the new species were significantly larger than those of M. mixtura(P 0.01), and the ratios of SL, IOD, LAHL, HLL, TL, TFL and FL to SVL of the new species were significantly less than those of M. mixtura(P 0.05).     

A New Species of Microhyla(Amphibia: Anura: Microhylidae) from Langbian Plateau,Central Vietnam

We describe a new species, Microh yla hongiaoensis sp. nov., from Lam Dong Province, southern Vietnam based on morphological data and molecular evidences. The new species is sister to M. pulchella by molecular phylogenetics and also most closely resembles M. pulchella in morphological characteristics, albeit differs from its congeners by a combination of the following morphological features:(1) size medium(SVL 13.6–14.7 mm in males and 18.3–18.6 mm in females);(2) fingers II–IV with small disks, dorsal surface of disks, without median longitudinal groove;(3) webbing formula I1?– 2 II1 –2 III1 – 2?IV2?– 1 V;(4) toe disks with dorsal median longitudinal groove;(5) dorsal back without two small black spots;(6) one small black spot adjacent behind the eyes;(7) few small black scapular spots in the flanks-belly and inguinal region;(8) palm with two small metatarsal tubercles;(9) tibiotarsal reaching beyond snout. M.hongiaoensis sp. nov. occurs in evergreen montane tropical forests at an elevation of 1500 m a.s.l.     

Systematic relationships of Oriental tiny frogs of the family Microhylidae (Amphibia, Anura) as revealed by mtDNA genealogy

We estimated the genealogical relationships and assessed systematic relationships among 45 out of 89 named species and four unnamed taxa from 11 of 14 genera of the Oriental microhylids from 1767 bp sequences of the mitochondrial DNA genes 12S rRNA and 16S rRNA using maximum parsimony, maximum likelihood, and Bayesian inference methods. Monophyly was rejected for the subfamily Microhylinae, and our data reveal four well-supported clades whose relationships to each other are unresolved: (A) Microhyla, Calluella, and Glyphoglossus, (B) Chaperina, (C) Kaloula, Phrynella, and Metaphrynella, and (D) Micryletta. They were genetically as divergent from each other as from another Oriental subfamily Kalophryninae, and could be recognized as distinct subfamilies. Within Clade A, our data reveal three well-supported subclades whose relationships to each other are unresolved: (AI) Microhyla-I, (AII) Calluella and Glyphoglossus, and (AIII) Microhyla-II. Of the two enigmatic Malaysian genera, whose subfamilial placement has been undetermined, Phrynella was found to be the sister species of Metaphrynella in Clade C, whereas Gastrophrynoides was grouped in the Papua-Australian subfamily Asterophryinae. Currently recognized subgenera and species groups within Microhyla based on morphology were not supported phylogenetically, and require thorough reassessments.     

印尼新几内亚巴布亚省豕蛙属4新种(两栖纲:姬蛙科)(英文)

姬蛙科豕蛙属(Choerophryne)已知5种,几乎都分布于新几内亚岛东部地区,已有物种的描述多基于较少数量的标本,由于标本贫乏,迄今无 后续研究.本文针对采集于1998-2003年的50号标本,对该属进行了再研究,并描述了分布 于新几内亚岛的巴布亚西北地区鲜为人知的4新种.与同属已知物种比较,新种的有效性得 到来自形态、声谱以及分子数据的综合分析结果的支持.在新几内亚岛西部地区豕蛙属4新 种中,至少有3种的种群密度很高,从而极大增加了对该属的了解     

Taxonomic relationships within the pan-oriental narrow-mouth toad Microhyla ornata as revealed by mtDNA analysis (Amphibia, Anura, Microhylidae)

A molecular phylogenetic survey was conducted using mtDNA sequences of 12S and 16S rRNA, and cyt-b genes to examine taxonomic relationships among populations of the Pan-Oriental microhylid, Microhyla ornata, from India, Bangladesh, Thailand, Laos, China, Taiwan, and the Ryukyu Archipelago of Japan. Two discrete clades are recognized within this species, one consisting of populations from India and Bangladesh, and the other encompassing the remaining populations. In the latter clade, populations from the Ryukyu Archipelago are clearly split from the rest (populations from Taiwan and the continent) with considerable degrees of genetic differentiations. Each of the three lineages is judged to represent a good species, and the name Microhyla ornata is restricted to the South Asian populations. For the populations from Taiwan and a wide region from China to Southeast Asia, the name Microhyla fissipes should be applied, whereas the Ryukyu populations are most appropriately referred to as Microhyla okinavensis, although further substantial genetic differentiations are recognized among some island group populations within this last species.     

Population estimation in the Torrent frog, Amolops lamtensis (Anura: Ranidae)

The results of four attempts at yearly intervals to estimate populations of Amolops lamtensis (Blgr.) along 360 m of forest stream are discussed and the most consistent estimates are obtained using Jolly's (1965) method of analysing such data. Observations of movement within the population indicate that most frogs are recaptured close to the original point of capture although a few show considerable downstream movement, possibly as a result of being swept away by the current. The recapture expectations are not, however, random and there is a strong indication that larger frogs are more liable to recapture. There is also evidence that frogs are available for recapture for different lengths of time, and it is likely that this leads to an under-estimate of the total population.