Raoul H. Bain

Cryptic Species of a Cascade Frog from Southeast Asia: Taxonomic Revisions and Descriptions of Six New Species

Abstract The Southeast Asian cascade frog, Rana livida (Blyth, 1856), has long been suspected to be a complex of species. Several different forms are described from across its vast range. The loss of type material and disparate sampling efforts are challenges. Is variability in this species due to geographic variation or to the presence of multiple species? We use concordant evidence from morphology, morphometrics, cellular DNA content, and allozyme electrophoresis to investigate diversity in R. livida from Vietnam. Three distinct species are recognized on the basis of morphology, as are four other suspect groups (morphotypes). Discriminant function analyses of morphometric data detect patterns of morphological variation among all seven groups. Pairwise comparison of cellular DNA content using t-tests shows significant differences among sympatric morphotypes, suggesting they represent distinct species. This hypothesis is supported by an analysis of 14 allozymic loci, in which fixed allelic differences are found among specimens in sympatry and allopatry. Examination of available type material of four junior synonyms of R. livida results in their recognition as species. One of these species, R. chloronota, is a wide-ranging species erroneously referred to as R. livida. Seven species occur in Vietnam. We describe six new cryptic species belonging to the Rana chloronota complex, redescribe R. chloronota, R. livida, R. sinica, and R. graminea, and give comments on R. leporipes. Three of these new species (R. bacboensis, new species, R. hmongorum, new species, and R. daorum, new species) occur in montane forests in northern Vietnam, and two (R. banaorum, new species and R. morafkai, new species) are known only from the Tay Nguyen Plateau of Vietnams Central Highlands. One species, R. megatympanum, new species, occurs in portions of both northern Vietnam and the Central Highlands. An identification key for the Rana chloronota complex from Vietnam is provided. The finding of six cryptic species within a small portion of the geographic region of R. chloronota suggests that many more cascade ranids await discovery. This documentation has serious implications for conservation; each of the new species occurs in sympatry with at least one other member of the complex. Consequently, far more species are being affected by habitat loss than was previously thought.

Molecular phylogeny of Rhacophoridae (Anura): A framework of taxonomic reassignment of species within the genera Aquixalus, Chiromantis, Rhacophorus, and Philautus

Phylogenetic relationships among representative species of the family Rhacophoridae were investigated based on 2904bp of sequences from both mitochondrial (12S rRNA, 16S rRNA, the complete t-RNA for valine), and nuclear (tyrosinase, rhodopsin) genes. Maximum parsimony, maximum likelihood, and Bayesian analyses were employed to reconstruct the phylogenetic trees. This analysis, combined with previous phylogenetic studies, serves as a framework for future work in rhacophorid systematics. The monophyly of Rhacophorus is strongly confirmed except for the species R.hainanus, which is the sister taxon to A.odontotarsus. The non-monophyly of the newly designated genus Aquixalus by Delorme et al. [Delorme, M., Dubois, A., Grosjean, S., Ohler, A., 2005. Une nouvelle classification générique et subgénérique de la tribu des Philautini (Amphibia, Anura, Ranidae, Rhacophorinae). Bull. Mens. Soc. Linn. Lyon 74, 165-171] is further confirmed. Aquixalus (Aquixalus) forms a well-supported monophyletic group within Kurixalus, whereas, Aquixalus (Gracixalus) is more closely related to species of Rhacophorus, Polypedates, and Chiromantis. Philautus as currently understood, does not form a monophyletic group. Philautus (Kirtixalus) is the sister group to the clade comprising Kurixalus and Aquixalus (Aquixalus), and more remotely related to Philautus (Philautus). Chiromantisromeri does not cluster with species of Chiromantis, and forms a basal clade to all rhacophorids save Buergeria. We propose some taxonomic changes that reflect these findings, but further revision should await more detailed studies, which include combined morphological and molecular analyses, with greater species sampling.

Herpetofaunal Diversity of Ha Giang Province in Northeastern Vietnam, with Descriptions of Two New Species

Abstract In April and May of 2000, herpetological surveys of Ha Giang Province, Vietnam, near the Chinese border were undertaken. Surveys concentrated on isolated forests of Mount Tay Con Linh II (contiguous with the highest peak in eastern Vietnam, Mt. Tay Con Linh). The 26-day survey yielded 36 species of amphibians and 16 species of reptiles. The collection contains elements of Himalayan as well as Indo-Malayan assemblages and documents a new country record (Philautus rhododiscus), eight new records east of the Red River (Bombina microdeladigitora, Megophrys parva, Amolops chapaensis, Chaparana delacouri, Chirixalus gracilipes, Philautus odontotarsus, Polypedates dugritei, Rhacophorus hoanglienensis), seven species complexes (Fejervarya limnocharis, Hoplobatrachus rugulosus, Limnonectes kuhlii, Rana chloronota, R. maosonensis, Polypedates dugritei, and P. leucomystax), three unidentified amphibian species, and two previously undescribed species of cascade ranid (Rana iriodes, new species and Rana tabaca, new species). Rana iriodes differs from R. daorum and Amolops chunganensis in having an iridescent green-gold dorsum, orange-red dorsolateral folds, a gold-white flank spot, vomerine teeth, and webbing to toe disks. Rana tabaca, new species, can be differentiated from other cascade ranids by a combination of characters: mottled brown upper lip, gold lip line below the eye to the arm insertion, shagreened dorsum, dorsolateral folds, and unpigmented eggs. Species accumulation curves indicate that the diversity of the region is still underestimated, which, along with the relatively extensive remnant forest, underscores the importance for a greater faunal understanding and conservation effort for the montane forests of the region.

Impending conservation crisis for Southeast Asian amphibians

With an understudied amphibian fauna, the highest deforestation rate on the planet and high harvesting pressures, Southeast Asian amphibians are facing a conservation crisis. Owing to the overriding threat of habitat loss, the most critical conservation action required is the identification and strict protection of habitat assessed as having high amphibian species diversity and/or representing distinctive regional amphibian faunas. Long-term population monitoring, enhanced survey efforts, collection of basic biological and ecological information, continued taxonomic research and evaluation of the impact of commercial trade for food, medicine and pets are also needed. Strong involvement of regional stakeholders, students and professionals is essential to accomplish these actions.

Is The Amphibian Tree of Life really fatally flawed

Wiens (2007 , Q. Rev. Biol. 82, 55–56) recently published a severe critique of Frost et al.s (2006, Bull. Am. Mus. Nat. Hist. 297, 1–370) monographic study of amphibian systematics, concluding that it is “a disaster” and recommending that readers “simply ignore this study”. Beyond the hyperbole, Wiens raised four general objections that he regarded as “fatal flaws”: (1) the sampling design was insufficient for the generic changes made and taxonomic changes were made without including all type species; (2) the nuclear gene most commonly used in amphibian phylogenetics, RAG‐1, was not included, nor were the morphological characters that had justified the older taxonomy; (3) the analytical method employed is questionable because equally weighted parsimony “assumes that all characters are evolving at equal rates”; and (4) the results were at times “clearly erroneous”, as evidenced by the inferred non‐monophyly of marsupial frogs. In this paper we respond to these criticisms. In brief: (1) the study of Frost et al. did not exist in a vacuum and we discussed our evidence and evidence previously obtained by others that documented the non‐monophyletic taxa that we corrected. Beyond that, we agree that all type species should ideally be included, but inclusion of all potentially relevant type species is not feasible in a study of the magnitude of Frost et al. and we contend that this should not prevent progress in the formulation of phylogenetic hypotheses or their application outside of systematics. (2) Rhodopsin, a gene included by Frost et al. is the nuclear gene that is most commonly used in amphibian systematics, not RAG‐1. Regardless, ignoring a study because of the absence of a single locus strikes us as unsound practice. With respect to previously hypothesized morphological synapomorphies, Frost et al. provided a lengthy review of the published evidence for all groups, and this was used to inform taxonomic decisions. We noted that confirming and reconciling all morphological transformation series published among previous studies needed to be done, and we included evidence from the only published data set at that time to explicitly code morphological characters (including a number of traditionally applied synapomorphies from adult morphology) across the bulk of the diversity of amphibians (Haas, 2003, Cladistics 19, 23–90). Moreover, the phylogenetic results of the Frost et al. study were largely consistent with previous morphological and molecular studies and where they differed, this was discussed with reference to the weight of evidence. (3) The claim that equally weighted parsimony assumes that all characters are evolving at equal rates has been shown to be false in both analytical and simulation studies. (4) The claimed “strong support” for marsupial frog monophyly is questionable. Several studies have also found marsupial frogs to be non‐monophyletic. Wiens et al. (2005, Syst. Biol. 54, 719–748) recovered marsupial frogs as monophyletic, but that result was strongly supported only by Bayesian clade confidence values (which are known to overestimate support) and bootstrap support in his parsimony analysis was < 50%. Further, in a more recent parsimony analysis of an expanded data set that included RAG‐1 and the three traditional morphological synapomorphies of marsupial frogs, Wiens et al. (2006, Am. Nat. 168, 579–596) also found them to be non‐monophyletic. Although we attempted to apply the rule of monophyly to the naming of taxonomic groups, our phylogenetic results are largely consistent with conventional views even if not with the taxonomy current at the time of our writing. Most of our taxonomic changes addressed examples of non‐monophyly that had previously been known or suspected (e.g., the non‐monophyly of traditional Hyperoliidae, Microhylidae, Hemiphractinae, Leptodactylidae, Phrynobatrachus, Ranidae, Rana, Bufo; and the placement of Brachycephalus within “Eleutherodactylus”, and Lineatriton within “Pseudoeurycea”), and it is troubling that Wiens and others, as evidenced by recent publications, continue to perpetuate recognition of non‐monophyletic taxonomic groups that so profoundly misrepresent what is known about amphibian phylogeny.

A Biogeographic Synthesis of the Amphibians and Reptiles of Indochina

ABSTRACT Indochina (Laos, Cambodia, and Vietnam) houses over 600 species of amphibians and reptiles, roughly a quarter of which has been described within the last 15 years. Herein, we undertake the first biogeographic synthesis of the regional herpetofauna since the first half of the 20th century. We review the literature to measure and map species richness and endemism, the contributions of regional faunas, and ecological characteristics of Indochinas amphibians (Anura, Caudata), and reptiles (Serpentes, Sauria, Testudines, Crocodylia). Dividing Indochina into 19 subregions defined by topography and geology, we estimate the similarity among the regional faunas and appraise the effects of area and survey effort on these comparative analyses. Variation in species composition is broadly correlated with topography, habitat complexity, and proximity to regions outside Indochina. Indochinas herpetofauna is dominated (in decreasing order) by endemic species, widely distributed species, a South China fauna, and a biota centered in Thailand and Myanmar. Species richness is highest in amphibians and snakes, and peaks in upland forests. Endemism, highest among amphibians and lizards, also peaks in forests of the regions northern uplands and Annamite Range. Endemic species occupy a narrower range of habitats than nonendemics. Patterns of richness and endemism are partially explained by ecological constraints: amphibians and lizards are more restricted to forests than snakes, turtles, and crocodiles; amphibians are more restricted to uplands, turtles to lowlands. We also assess biogeography in the context of Indochinas geology, climate, and land cover. In northern Indochina, the Red River either acts as or coincides with an apparent dispersal barrier. Herpetofauna in northeastern upland areas are closely allied with fauna of southeastern China. In southern Indochina there is little evidence that the Mekong River represents a biogeographic barrier to the regional herpetofauna. The Annamite Range is composed of at least three distinct units and its elevated species richness and endemism are also noted in adjacent lowlands. Contribution of subtropical biota to Indochinas fauna is significantly greater than that of tropical biota and there is little other evidence for intermixing at intermediate latitudes. Our results have implications for biogeography and conservation efforts, although they must be viewed in the context of rapidly evolving systematic knowledge of the regions amphibians and reptiles. Future survey efforts, and the phylogenetic analyses that come from them, are essential for supporting regional conservation efforts, as they will better resolve the known patterns of amphibian and reptile richness and endemism.

Three New Indochinese Species of Cascade Frogs (Amphibia: Ranidae) Allied to Rana archotaphus

Abstract Three new frog species allied to Rana archotaphus are described from Laos and Vietnam. One new species from the Hoang Lien Mountains of northwestern Vietnam is characterized by having females with snout-vent length 66–68 mm, no visible pineal body, the first finger shorter than second, width of third finger disc about equal to tympanum diameter, and tympanum of males relatively larger than females. The second new species from northern Laos is characterized by having males with snout-vent length 38–44 mm, no visible pineal body, first finger shorter than second, width of third finger disc equal to the tympanum diameter, venter with very light or no spotting, and venter translucent. The third new species from the Northern Truong Son (Annamite Highlands) of Laos and the Kon Tum Plateau of Vietnam is characterized by having females with snout-vent length 56–57 mm, a visible pineal body, tympanum of males relatively equal to females, the first finger shorter than second, and males with width of third finger disc about half the tympanum diameter. Descriptions are expanded of several species that resemble the two new species: Rana archotaphus, R. daorum, R. iriodes, and Amolops chunganensis.

The stream-dwelling tadpole of Hyloscirtus charazani (Anura: Hylidae) from Andean Bolivia

Abstract We describe the lotic larva of Hyloscirtus charazani from Charazani and nearby areas, Departamento La Paz, Bolivia (2700–3200 m a.s.l.). It is morphologically undistinguishable from tadpoles of H. armatus sensu lato. These tadpoles show stream adaptation through a robust body with strong tail musculature, enlarged oral disc, increased number of labial tooth rows, and complete marginal papillae. Furthermore, they have a large saccular structure that underlies the limb buds, encloses the vent tube, and partially covers the hindlimbs during their development. Resumen Describimos la larva lótica de Hyloscirtus charazani de Charazani y de otra localidad cercana, Departamento La Paz, Bolivia (2700–3200 m sobre el nivel del mar). Morfologicamente es identica con renacuajos de H. armatus sensu lato. Estos renacuajos muestran caracteres comunmente entendidos como adaptaciones a la vida en arroyos rapidos, como ser el cuerpo robusto, musculatura de la cola robusta, disco oral ensanchado, un aumento en las filas de dientes labiales y papila marginal completa. Ademas, tienen una estructura sacular grande, bajo los primordios de los miembros posteriores, que contiene al tubo cloacal, y que envuelve parcialmente a los miembros durante su desarrollo.

Three New Species of Narrow-Mouth Frogs (Genus: Microhyla) from Indochina, with Comments on Microhyla annamensis and Microhyla palmipes

Abstract Three species of Microhyla from Indochina are described. One species is from the Northern Truong Son (Annamite Highlands) of Vietnam and Laos, characterized by finger I less than one-half the length of finger II; expanded disks at the tip of the digits with dorsal median longitudinal grooves producing the appearance of two scutes; two metatarsal tubercles; extensive webbing on feet: on toe IV web full to distal subarticular tubercle; and marbled belly. The second species is from the Tay Nguyen Plateau, Gia Lai Province, Vietnam. It is smaller than the above species, has full webbing to medial subarticular tubercle of toe IV, and has a dusted belly. Microhyla annamensis is discussed because of its similarity to the two new species above. A third species of Microhyla is described from Ngoc Linh Mountain in Quang Nam Province, Vietnam. It is easily separated from congeners of Indochina and China by its small size and the appearance of the first finger as a small, partially free nub. Microhyla palmipes is discussed because of its similarity to the third new species.

THREE NEW SPECIES OF SPINULE-BEARING FROGS ALLIED TO RANA MEGATYMPANUM BAIN, LATHROP, MURPHY, ORLOV AND HO, 2003 FROM LAOS AND VIETNAM

Three new species of frogs allied to Rana megatympanum are described from Laos and Vietnam. One new species is from the Bolaven Plateau of southern Laos, a second is from central Laos and adjacent central Vietnam, and a third is from northern Laos. The three new species are distinguished from similar species by skin texture, the location of white spinules on the skin in males and females, spotting on the lips, body size, relative tympanum size, and unpigmented eggs.