Female sperm storage in amphibians.

The three orders of extant amphibians are Gymnophiona, Anura, and Urodela. Although all gymnophionans apparently have internal fertilization and many are viviparous, female sperm storage is unknown. Internal fertilization has convergently evolved in a few anurans, but females of just one species, Ascaphus truei, are known to possess oviductal sperm storage tubules (SSTs). The SSTs of A. truei are similar anatomically to such glands in squamate reptiles. This similarity is convergence due to similar functional adaptations and/or internal design constraints. In salamanders and newts (Urodela), absence of sperm storage in females is the ancestral condition (three families). In the derived condition, sperm storage occurs in cloacal glands called spermathecae, and their possession is a synapomorphy for females in the suborder Salamandroidea (seven families). Salamandroids are the only vertebrates with cloacal sperm storage glands. In this paper, a phenetic analysis of variation in spermathecal characters reveals patterns of convergence in certain spermathecal characters in unrelated taxa that breed in similar habitats. In the family Salamandridae, a role in sperm nutrition for the spermathecal epithelium is questioned, and the widespread occurrence of spermiophagy is related to other reproductive strategies. I propose how the packaging of sperm in structurally different types of spermathecae may influence male paternity.     

两栖动物线粒体基因组结构特征分析

对2007年6月13日以前公布于GenBank上的78种两栖动物的线粒体基因组全序列进行了总结、比较和分析。78种基因组中基因的数量从35~41个不等;根据基因的数量、种类及其排列顺序的差别将其分为22种基因组类型,其进而聚为3组,其中类型4为两栖纲与其它脊椎动物的常见类型,类型8为两栖纲中现生3个目的公有类型。与类型4比较,其余21种线粒体基因组类型涉及基因变动的基因共有18个,其中变动比较多的是tRNA基因,移位、增多和缺失的发生频率都较大,而蛋白编码基因比较稳定,主要是移位。78种两栖动物中,蚓螈目的线粒体基因组均小于18000bps,多数在15000~16000bps;有尾目和无尾目均大于16000bps,其中有尾目多数在16000~17000bps,无尾目的多数在17000~18000bps。     

Amphibian biology and husbandry

Extant amphibians comprise three lineages-- salamanders (Urodela or Caudata), frogs and toads (Anura), and caecilians (Gymnophiona, Apoda, or Caecilia)--which contain more than 6,000 species. Fewer than a dozen species of amphibians are commonly maintained in laboratory colonies, and the husbandry requirements for the vast majority of amphibians are poorly known. For these species, a review of basic characteristics of amphibian biology supplemented by inferences drawn from the morphological and physiological characteristics of the species in question provides a basis for decisions about housing and feeding. Amphibians are ectotherms, and their skin is permeable to water, ions, and respiratory gases. Most species are secretive and, in many cases, nocturnal. The essential characteristics of their environment include appropriate levels of humidity, temperature, and lighting as well as retreat sites. Terrestrial and arboreal species require moist substrates, water dishes, and high relative humidity. Because temperature requirements for most species are poorly known, it is advisable to use a temperature mosaic that will allow an animal to find an appropriate temperature within its cage. Photoperiod may affect physiology and behavior (especially reproduction and hibernation), and although the importance of ultraviolet light for calcium metabolism by amphibians is not yet known, ecological observations suggest that it might be important for some species of frogs. Some amphibians are territorial, and some use olfactory cues to mark their territory and to recognize other individuals of their species. All amphibians are carnivorous as adults, and the feeding response of many species is elicited by the movement of prey. Diets should include a mixture of prey species, and it may be advisable to load prey with vitamins and minerals.     

Initial diversification of living amphibians predated the breakup of Pangaea

The origin and divergence of the three living orders of amphibians (Anura, Caudata, Gymnophiona) and their main lineages are one of the most hotly debated topics in vertebrate evolution. Here, we present a robust molecular phylogeny based on the nuclear RAG1 gene as well as results from a variety of alternative independent molecular clock calibrations. Our analyses suggest that the origin and early divergence of the three living amphibian orders dates back to the Palaeozoic or early Mesozoic, before the breakup of Pangaea, and soon after the divergence from lobe-finned fishes. The resulting new biogeographic scenario, age estimate, and the inferred rapid divergence of the three lissamphibian orders may account for the lack of fossils that represent plausible ancestors or immediate sister taxa of all three orders and the heretofore paradoxical distribution of some amphibian fossil taxa. Furthermore, the ancient and rapid radiation of the three lissamphibian orders likely explains why branch lengths connecting their early nodes are particularly short, thus rendering phylogenetic inference of implicated relationships especially difficult.     

Evolutionary morphology of the caecilian urogenital system. I. The gonads and the fat bodies

The comparative morphology of the gonads and fat bodies of members of 17 genera and 46 species of caecilians (Amphibia: Gymnophiona) is described and analyzed. Comparison is made with the morphology of salamanders and frogs in order to elucidate evolutionary trends and relationships within the order Gymnophiona and within the class Amphibia. The structure of the testis lobes and transverse and longitudinal ducts is described based on gross dissection and histological investigation. The pattern of spermatogenesis and interstitial tissue changes are described and compared with those of other amphibians. A trend toward fusion of testis lobes is analyzed. The characteristics of the seasonal reproductive cycle of male Gymnopis m. proxima are described, and evidence for cyclic reproductive activity in other forms is presented. The morphology of the ovaries and ova is described. Size of ovary and size and number of ova is dependent on the state of maturation of the ova. Some evidence for seasonal ovum production and breeding is presented. Fat body morphology is found to be correlated with size, nutrition, and gonad condition, as in other amphibians.     

Remarkable genetic homogeneity in unstriped, long-tailed Ichthyophis along 1500 km of the Western Ghats, India

The mountainous Western Ghats are part of a biodiversity hotspot and extend for about 1600 km, lying close and subparallel to the west coast of peninsular India. The region is a centre of diversity for amphibians, and recent preliminary work on some components of both caecilian (Gymnophiona) and frog (Anura) fauna is indicative of a high degree of local endemism. We investigated diversity in mitochondrial rRNA 12S and 16S sequences for long-tailed, unstriped Ichthyophis (Amphibia: Gymnophiona: Ichthyophiidae) that potentially represent four taxonomically confused and poorly known endemic caecilian species. Data were analysed for18 individuals from along c . 1500 km of the Western Ghats region. Genetic diversity is remarkably low, with a maximum uncorrected p -distance of 0.5%. These DNA sequences and new morphological data do not allow us to reject the null hypothesis that the sample comprises only a single, widely distributed, highly interconnected species. The phylogenetic signal among the data is extremely low. However, population genetic analyses reveal that the Palghat Gap, a c . 30 km discontinuity in the Western Ghats considered to be significant in the biogeography of other organisms occurring in this region, corresponds to a significant subdivision of long-tailed, unstriped Ichthyophis into two groups.     

探讨两栖纲三个目之间的系统发生关系

现存两栖纲分为3个目,然而它们的系统发生关系仍存在较大的争议.选择Genbank上25种两栖动物线粒体全序列,结合已测定的福建大头蛙(Lim nonectes fujianensis)线粒体基因组全序列,以天鹅和眼镜凯门鳄为外群,用线粒体tRNA基因的合并数据重建系统树.采用P AU P(version 4.0b10)软件构建MP和NJ树,tree-puzzle5.2构建ML树.结果显示,NJ树和ML树以较高的自引导值支持有尾目和蚓螈目为姐妹群,而MP树则支持无尾目和有尾目为姐妹群.     

Pterygoideus muscles and other jaw adductors in amphibians and reptiles

The general structural patterns of jaw adductors in all orders of extant amphibians and reptiles, and also polypteriforms, crossopterygians (coelacanth), and dipnoans, are compared. The pterygoideus muscles probably developed independently and in parallel in gymnophions and amniotes from the profound pseudotemporalis muscle, which was present in their fishlike ancestors and was retained in caudate and anuran amphibians. The functional causes of the development of pterygoideus muscles in the majority of tetrapod groups and the absence of these muscles in Urodela and Anura are discussed. The anterior pterygoideus muscle of crocodiles is homologous to the pseudotemporalis (superficial) muscle of other reptiles.     

用线粒体tRNA基因探讨现存两栖动物三个目间系统发生关系

现存两栖类3个目的系统发生关系仍然没有统一意见,最广泛被接受的假说是单系起源,并且无尾类和有尾类为姐妹群关系而排斥蚓螈类(蛙类假说)。然而,这一假说一直存在争议。我们在测定了泽蛙线粒体基因组全序列的基础上,与已知其他的6种两栖类进行详细的比较分析,同时选择了11种高等脊椎动物的线粒体全基因序列,以硬骨鱼类作外群,用22个tRNA基因合并数据进行系统发生重建分析,结果表明MP、ML树都强力地支持现生两栖类动物为单系群,并且有尾目和蚓螈目为姐妹群关系。这个结果与蛙类假说是相矛盾的,与Bolt(1991)在形态学基础上提出的有尾类和蚓螈类为姐妹群关系的假说相一致,并得到建立在线粒体和核rRNA基因数据基础上的许多分子研究的支持。另外还探讨了本结果与前人的研究不一致的原因,以及利用线粒体全基因序列进行系统发生分析可能存在的偏差。     

中国两栖、爬行动物更新名录

本文在2015年发表的爬行动物名录及同年《中国两栖类信息系统》发布的两栖动物名录的基础上, 通过整理新发表的分类学研究及先前名录遗漏的部分早期文献, 更新了截至2019年底中国现生本土两栖、爬行动物物种名录。2015-2019年间, 中国两栖动物新记录1科, 新描述2属, 恢复1属有效性, 新记录1属, 新描述或恢复有效种74种, 新增国家纪录18种; 另6属、8种的有效性未得到近年研究证据支持(在此视为次定同物异名而未做收录, 后同)。同期, 中国爬行动物新恢复5科, 新描述1亚科, 新描述1属, 恢复3属有效性, 新记录3属, 新描述、恢复或提升有效种43个, 新增国家纪录10种; 另有5属、4种的有效性未得到近年研究证据支持, 并移除1属、4种在我国的分布纪录。此外, 通过整理2015年前文献, 爬行动物增补3属, 提升3亚种至种级地位, 增补国家新纪录3种, 另有3属、2种的有效性未得到近年研究证据支持, 同时移除1种在我国的分布纪录。综上, 截至2019年底, 我国共记录现生本土两栖动物3目13科62属515种(蚓螈目1科1属1种, 有尾目3科14属82种, 无尾目9科47属431种), 爬行动物3目35科135属511种(鳄形目1科1属1种, 龟鳖目6科18属34种, 有鳞目蛇亚目18科73属265种、蜥蜴亚目10科43属211种)。此外, 本文还对先前名录中部分爬行动物的中文名提出了修改建议, 建议恢复部分物种的惯用中文名。2015-2019年, 新物种及新纪录已知物种数量占现两栖、爬行动物物种总数的17.1%和10.2%。近年来, 我国发表的两栖、爬行动物新物种和已知物种的新纪录数量持续增加, 分类体系也在研究中不断完善, 建议今后及时地进行阶段性总结, 同时对存在的问题提出讨论, 以推动中国两栖、爬行动物分类学研究工作的进一步开展。     

Phylogeny of caecilian amphibians (Gymnophiona) based on complete mitochondrial genomes and nuclear RAG1

We determined the complete nucleotide sequence of the mitochondrial (mt) genome of five individual caecilians (Amphibia: Gymnophiona) representing five of the six recognized families: Rhinatrema bivittatum (Rhinatrematidae), Ichthyophis glutinosus (Ichthyophiidae), Uraeotyphlus cf. oxyurus (Uraeotyphlidae), Scolecomorphus vittatus (Scolecomorphidae), and Gegeneophis ramaswamii (Caeciliidae). The organization and size of these newly determined mitogenomes are similar to those previously reported for the caecilian Typhlonectes natans (Typhlonectidae), and for other vertebrates. Nucleotide sequences of the nuclear RAG1 gene were also determined for these six species of caecilians, and the salamander Mertensiella luschani atifi. RAG1 (both at the amino acid and nucleotide level) shows slower rates of evolution than almost all mt protein-coding genes (at the amino acid level). The new mt and nuclear sequences were compared with data for other amphibians and subjected to separate and combined phylogenetic analyses (Maximum Parsimony, Minimum Evolution, Maximum Likelihood, and Bayesian Inference). All analyses strongly support the monophyly of the three amphibian Orders. The Batrachia hypothesis (Gymnophiona, (Anura, Caudata) receives moderate or good support depending on the method of analysis. Within Gymnophiona, the optimal tree (Rhinatrema, (Ichthyophis, Uraeotyphlus), (Scolecomorphus, (Gegeneophis Typhlonectes) agrees with the most recent morphological and molecular studies. The sister group relationship between Rhinatrematidae and all other caecilians, that between Ichthyophiidae and Uraeotyphlidae, and the monophyly of the higher caecilians Scolecomorphidae+Caeciliidae+Typhlonectidae, are strongly supported, whereas the relationships among the higher caecilians are less unambiguously resolved. Analysis of RAG1 is affected by a spurious local rooting problem and associated low support that is ameliorated when outgroups are excluded. Comparisons of trees using the non-parametric Templeton, Kishino-Hasegawa, Approximately Unbiased, and Shimodaira-Hasegawa tests suggest that the latter may be too conservative.     

Mitochondrial evidence on the phylogenetic position of caecilians (Amphibia: Gymnophiona)

The complete nucleotide sequence (17,005 bp) of the mitochondrial genome of the caecilian Typhlonectes natans (Gymnophiona, Amphibia) was determined. This molecule is characterized by two distinctive genomic features: there are seven large 109-bp tandem repeats in the control region, and the sequence for the putative origin of replication of the L strand can potentially fold into two alternative secondary structures (one including part of the tRNA(Cys)). The new sequence data were used to assess the phylogenetic position of caecilians and to gain insights into the origin of living amphibians (frogs, salamanders, and caecilians). Phylogenetic analyses of two data sets-one combining protein-coding genes and the other combining tRNA genes-strongly supported a caecilian + frog clade and, hence, monophyly of modern amphibians. These two data sets could not further resolve relationships among the coelacanth, lungfishes, and tetrapods, but strongly supported diapsid affinities of turtles. Phylogenetic relationships among a larger set of species of frogs, salamanders, and caecilians were estimated with a mitochondrial rRNA data set. Maximum parsimony analysis of this latter data set also recovered monophyly of living amphibians and favored a frog + salamander (Batrachia) relationship. However, bootstrap support was only moderate at these nodes. This is likely due to an extensive among-site rate heterogeneity in the rRNA data set and the narrow window of time in which the three main groups of living amphibians were originated.     

Direct development in the tailed amphibians (salamanders), its origin and evolution

Biphasic life circle including water larvae and terrestrial adults is the primitive character for recent amphibians. Respectively, the larval development type (sensu Zakhvatkin, 1975) is also primitive for them. However, many amphibians possess direct development, in which the most part of ontogenesis takes place in the egg, and a miniature copy of the adult adapted to the terrestrial mode of life comes into the world. Transition from the larval type to the direct one occurs several times independently in Apoda, Urodela, and Anura. Pathways and mechanisms of formation of the direct type, its evolutionary tendencies, and ontogenetic prerequisites of that transition are studied in the plethodontid urodelans herewith in details. It is observed that the entire process of initial formation and subsequent specialization of the direct type involves: 1) progressing displacement of ontogenesis into embryogenesis and loss of larval characters, 2) desynchronization of initially metamorphic transformations and processes with their progressive lost, 3) acceleration of the beginning of functional activity of the thyroid gland, and 4) subsiding of the role of thyroid hormones in the ontogenesis regulation. Transition to the direct development type involves similar kinds of ontogenetic transformations and regulations in both Anura and Urodela despite of their independent evolution. Respectively, mechanisms of that transition are universal for the amphibians. The ontogenetic prerequisites of that transition are shown to be either significant dissociability of the larval and adult stages of ontogenesis (in anurans and plethodontid urodelans), or absence of the extreme larval specializations and respective sharp differences between larvae and adults (in extant Apoda and extinct labyrinthodonts and seymouriamorphs).     

猫儿山小鲵肝的组织结构

通过常规石蜡切片技术对2例猫儿山小鲵(Hynobius maoershanens)肝进行组织学观察。结果显示,猫儿山小鲵肝分为两叶,右叶稍大于左叶。肝组织结构主要由被膜、中央静脉、门管区和肝细胞组成。门管区的小叶间静脉和小叶间胆管清晰可见,但小叶间动脉不易观察。肝内结缔组织少,肝小叶之间界限不清。肝细胞索围绕中央静脉呈放射状排列,但放射状不明显。肝实质中含有大量清晰可见的棕黑色色素团,可能与此物种对低氧环境的适应有关。     

龙洞山溪鲵精子的超微结构

应用透射电镜和扫描电镜对龙洞山溪鲵(Batrachuperuslongdongensis)精子的超微结构进行观察和研究,探讨山溪鲵属以及小鲵科物种精子的结构特征,并探讨有尾两栖类精子结构特征的演化及其与生殖进化的关系。结果表明:1)龙洞山溪鲵的精子具小鲵科物种精子的共同特征,即精子无顶体钩,顶体呈三叶草状,尾部无线粒体,轴纤维粗大呈圆柱状等特征;2)龙洞山溪鲵精子核脊的形态结构与小鲵科其它属物种以及有尾类其它科物种精子的核脊存在明显的差别,由此可见核脊形态结构具有属或种的特异性。此外,与已有报道的有尾两栖类物种精子的结构特征进行分析比较表明:1)有尾类精子特征的分化与其受精方式的变化是一致的;2)精子结构特征支持隐鳃鲵超科为单系的推测     

Study of regeneration in amphibians in age of molecular-genetic approaches and methods

The results of molecular-genetic mechanisms of regeneration in amphibians are reviewed. Based on the examples of traditional and well-studied models of the restoration of the retinas and lenses of eyes, as well as limbs and tails in amphibians, we analyze the current state of regeneration problems and questions linked to cell reprogramming, growth, and morphogenesis. The development of the Kol’tsov school of thought in the age of molecular-genetic approaches and methods are monitored. The contemporary interpretation of organ regeneration in terms of molecular-genetic regulation and a new look at the definition of regeneration as repeated development is proposed. We also emphasize the current problems that exist in that field of developmental biology and are caused by the many difficulties of genome sequencing and the introduction of transgenesis in Urodela, the animal species with the highest regeneration abilities.     

The hyal and ventral branchial muscles in caecilian and salamander larvae: Homologies and evolution

Amphibians (Lissamphibia) are characterized by a bi‐phasic life‐cycle that comprises an aquatic larval stage and metamorphosis to the adult. The ancestral aquatic feeding behavior of amphibian larvae is suction feeding. The negative pressure that is needed for ingestion of prey is created by depression of the hyobranchial apparatus as a result of hyobranchial muscle action. Understanding the homologies of hyobranchial muscles in amphibian larvae is a crucial step in understanding the evolution of this important character complex. However, the literature mostly focuses on the adult musculature and terms used for hyal and ventral branchial muscles in different amphibians often do not reflect homologies across lissamphibian orders. Here we describe the hyal and ventral branchial musculature in larvae of caecilians (Gymnophiona) and salamanders (Caudata), including juveniles of two permanently aquatic salamander species. Based on previous alternative terminology schemes, we propose a terminology for the hyal and ventral branchial muscles that reflects the homologies of muscles and that is suited for studies on hyobranchial muscle evolution in amphibians. We present a discussion of the hyal and ventral branchial muscles in larvae of the most recent common ancestor of amphibians (i.e. the ground plan of Lissamphibia). Based on our terminology, the hyal and ventral branchial musculature of caecilians and salamanders comprises the following muscles: m. depressor mandibulae, m. depressor mandibulae posterior, m. hyomandibularis, m. branchiohyoideus externus, m. interhyoideus, m. interhyoideus posterior, m. subarcualis rectus I, m. subarcualis obliquus II, m. subarcualis obliquus III, m. subarcualis rectus II‐IV, and m. transversus ventralis IV. Except for the m. branchiohyoideus externus, all muscles considered herein can be assigned to the ground plan of the Lissamphibia with certainty. The m. branchiohyoideus externus is either apomorphic for the Batrachia (frogs + salamanders) or salamander larvae depending on whether or not a homologous muscle is present in frog tadpoles. J. Morphol., 2011. © 2011 Wiley‐Liss, Inc.     

The determination of ectodermal derivatives in various species of amphibia

The process of labyrinth determination has been studied in amphibians (three Urodela and seven Anura species) using homoplastic transplantation of ear ectoderm, containing labyrinth material onto the abdominal wall of embryos of the same stage of development. The stage of appearance of organ-specific properties in ear ectoderm was determined and the increase of these properties in the course of development was observed. The frequency of ear vesicles formation, the level of their differentiation and their size served as criteria. These criteria allow to align the studied species into a row, where organ-specific properties appear earlier and most completely in representatives of Ranidae family and in Bufo viridis, and weakest, in smooth newt. A comparison of properties of labyrinth material and other areas of ectoderm allowed to conclude that specific differences in determination of different ectodermal primordia are based on specific peculiarities of the whole ectoderm. The appearance of these differences can be explained by the shift in the beginning of gastrulation towards later stages of cleavage during the evolution of amphibians.     

中国两栖动物受威胁现状评估

为了了解我国两栖动物受威胁现状和致危因素, 进而制定相关的保护措施和开展国际合作, 本文依据中国两栖动物野生种群与生境现状, 利用《IUCN物种红色名录濒危等级和标准》(3.1版)和《IUCN物种红色名录标准在国家或地区的应用指南》(4.0版), 对中国已知的408种两栖动物的濒危状况进行了评估, 并编制了《中国两栖动物红色名录》。评估结果表明: 中国两栖动物有1种灭绝, 1种区域灭绝, 受威胁的两栖动物共计176种, 占评估物种总数的43.1%, 明显高于《IUCN濒危物种红色名录》(2015)的物种受威胁率(30.8%)。中国两栖动物特有种272种, 其中48.9%属于受威胁物种。中国两栖动物受威胁比例最高的目是有尾目(63.4%), 明显高于无尾目(39.0%); 受威胁比例最高的科是隐鳃鲵科(Cryptobranchidae) (仅有1种, 100%受威胁), 小鲵科(Hynobiidae) (86.7%)和叉舌蛙科(Dicroglossidae) (78.1%)。有11个省区的受威胁物种数占本省区两栖动物物种总数的30%及以上, 前3位分别是四川(40.8%)、广西(39.2%)和云南(37%)。中国大多数两栖动物物种分布在西南山地和华南地区, 以海拔2,000 m以下区域为主。栖息地退化或丧失、捕捉、环境污染列受威胁两栖动物致危因子的前3位。鉴于中国两栖动物区系的复杂性和独特性, 进一步加强两栖动物资源调查、种群和生境监测及相关科学研究, 仍是今后一段时期开展两栖动物多样性保护和濒危物种拯救行动的关键性基础工作。     

Risk of extinction increases towards higher elevations across the world's amphibians

Aim

Life in mountains is associated with multiple features that increase the risk of demographic collapses in populations – small geographic ranges, short breeding seasons, specialization to harsh climates – leading to the hypothesis that extinction risk is exacerbated in species inhabiting higher elevations. Here, we implement the first test of this hypothesis across the amphibian tree of life – the tetrapods with the largest proportion of montane species, and nature's most threatened animals.

Location

Global.

Time Period

Present.

Major Taxa Studied

Class Amphibia.

Methods

We collated a dataset spanning 8042 species from across all three amphibian orders (Anura, Caudata and Gymnophiona). We preformed phylogenetic logistic regressions to test the predictions that extinction risk increases with elevation, and whether this effect is caused by factors previously hypothesised to drive high-elevation declines, including restrictions on species' geographic ranges, variation in their life histories and the presence of infectious disease.

Results

Globally, extinction risk increases towards higher elevations. At order-level, this relationship holds for frogs and salamanders. Even when controlling for geographic range size, life histories and infectious disease, extinction risk increases with elevation for amphibians combined and frogs globally, and in the Americas. In contrast, whereas extinction risk is greater among high-elevation Eurasian amphibians, this relationship is explained by larger body sizes and lower fecundity.

Main Conclusions

Our analyses indicate that after considering factors previously thought to explain the increase in extinction risk towards higher elevations (e.g., geographic range size, disease), elevation remains a significant predictor of amphibian extinction risk. Given that the only available tests of this hypothesis in other tetrapods (birds and reptiles) conflict with our findings, we suggest that physiological or life-history features of amphibians may explain this observed phenomenon.