A new species of the Gekko japonicus group (Squamata: Gekkonidae) from Khammouane Province, central Laos

A new species of the Gekko (Japonigekko) japonicus group from Khammouane Province, central Laos is described based on morphological characters and molecular evidence. Morphologically, Gekko khunkhamensis sp. nov. can be distinguished from the remaining congeners by a combination of the following characters: size moderate (SVL 69.7–75.2 mm); nares in contact with rostral; internasals absent; postmentals enlarged; interorbital scales between anterior corners of the eyes 31 or 32; dorsal tubercles absent; ventrals between mental and cloacal slit 181–185; midbody scales 127–138; ventral scales 42–45; subdigital lamellae on first toe 13 or 14, on fourth toe 14 or 15; tubercles on upper surface of fore and hind limbs absent; precloacal pores absent in the male and females; postcloacal tubercles 2; tubercles absent on dorsal surface of tail base; subcaudals distinctly enlarged; dorsal surface of body with five dark grey bands, which become irregular posteriorly. Genetically, the new species is placed in a clade, consisting of G. bonkowskii, G. nadenensis, G. scientiadventura, G. sengchanthavongi, and G. thakhekensis and differs from other congeners by at least 13% in terms of pairwise distance based on a fragment of the mitochondrial ND2 gene.

A New Species of Cyrtodactylus Gray, 1827 (Squamata, Gekkonidae) from Southern Yunnan, China

A new species of Cyrtodactylus is described based on five specimens collected from the karst formations of Maguan County, Wenshan Prefecture, Yunnan Province, China. The new species is recognized by having a unique combination of morphological characters: medium body size, ventrolateral folds present without interspersed tubercles, 7 – 9 precloacal pores in a continuous series, enlarged femoral scales present and continuous with pore-bearing precloacal scales, femoral pores on each enlarged femoral scale in males, 1 – 4 postcloacal tubercles on each side, subcaudals enlarged, a black postocular streak extending from posterior corner of eye rearwards to above tympanum, nuchal loop discontinuous, 6 – 7 black irregular dorsal bands between limbs, most bands discontinuous. Genetically, uncorrected sequence divergences of the ND2 gene and its flanking tRNAs between the new species and investigated congeners ranged from 12.5% to 18.2%.

The phylogenetic position of the enigmatic Assam day gecko Cnemaspis cf. assamensis (Squamata: Gekkonidae) demonstrates a novel biogeographic connection between Northeast India and south India-Sri Lanka

Northeast Indian biodiversity has long been considered to have a stronger affinity to Southeast Asian rather than Peninsular Indian fauna, however, few molecular phylogenetic studies have explored this hypothesis. In Asia, the polyphyletic gekkonid genus Cnemaspis sensu lato is comprised of two distantly related groups; one primarily from South Asia with some members in Southeast Asia, and the other exclusively from Southeast Asia. Cnemaspis assamensis is a systematically obscure and geographically isolated species (>1400 km from its nearest congeners) from the Brahmaputra River Valley in Northeast India. We provide the first molecular phylogenetic assessment of this species based on a partial ND2 gene fragment. Cnemaspis assamensis is determined to be a deeply divergent (Oligocene) member of the South Asian radiation and is sister to the podihuna clade which is endemic to Sri Lanka. The biogeographic implications of this find are discussed and this is suspected to represent a rare example of true disjunction between the wet zones of Northeast India and southern India/Sri Lanka. These results further emphasise the importance of Northeast India as a refuge for unique ancient faunal lineages.

Four new Bent-toed geckos (Cyrtodactylus Gray: Squamata: Gekkonidae) from northeast India

Based on morphology and ND2 gene sequences, four new species of Cyrtodactylus, two each from the Indian states of Meghalaya and Mizoram are described herein. The new species are a part of the Cyrtodactylus khasiensis group. The species from Meghalaya represent the lowland clade whereas the species from Mizoram represent the highland clade within the south of Brahmaputra clade of Indo-Burmese Cyrtodactylus. The two distinct populations from Meghalaya are sister to one another, differing from each by an uncorrected p-distance 0.065 and collectively are sister to Cyrtodactylus septentrionalis. The species from Mizoram differ from each other by an uncorrected p-distance of 0.085–0.121 and collectively are sister to Cyrtodactylus montanus.

Taxonomic reassessment and phylogenetic placement of Cyrtodactylus phuketensis (Reptilia, Gekkonidae) based on morphological and molecular evidence

The taxonomy and phylogeny of the Cyrtodactylus pulchellus complex along the Thai-Malay Peninsular region has been the focus of many recent studies and has resulted in the recognition of 17 species. However, the majority of these studies were focused on Peninsular and insular Malaysia where there were specimens and genetic vouchers. The taxonomic status and phylogenetic relationships of the Thai species in this complex remain unresolved, due to the lack of genetic material of some species, especially C. phuketensis and C. macrotuberculatus from Thai populations. In this study, we investigated the phylogenetic relationship between C. phuketensis and its closely related species C. macrotuberculatus, using both morphometric and molecular data. Phylogenetic analyses of mitochondrial NADH dehydrogenase subunit 2 (ND2) gene revealed that C. phuketensis is embedded within a C. macrotuberculatus clade with 1.45–4.20% (mean 2.63%) uncorrected pairwise sequence divergences. Morphological comparisons showed nearly identical measurements of C. phuketensis and C. macrotuberculatus and overlapping ranges in meristic characters. Based on these data, C. phuketensis is considered to be a variant of C. macrotuberculatus, thus rendering C. phuketensis a junior synonym of C. macrotuberculatus.

Interspecies Behavioral Variability of Medaka Fish Assessed by Comparative Phenomics

Recently, medaka has been used as a model organism in various research fields. However, even though it possesses several advantages over zebrafish, fewer studies were done in medaka compared to zebrafish, especially with regard to its behavior. Thus, to provide more information regarding its behavior and to demonstrate the behavioral differences between several species of medaka, we compared the behavioral performance and biomarker expression in the brain between four medaka fishes, Oryzias latipes, Oryzias dancena, Oryzias woworae, and Oryzias sinensis. We found that each medaka species explicitly exhibited different behaviors to each other, which might be related to the different basal levels of several biomarkers. Furthermore, by phenomics and genomic-based clustering, the differences between these medaka fishes were further investigated. Here, the phenomic-based clustering was based on the behavior results, while the genomic-based clustering was based on the sequence of the nd2 gene. As we expected, both clusterings showed some resemblances to each other in terms of the interspecies relationship between medaka and zebrafish. However, this similarity was not displayed by both clusterings in the medaka interspecies comparisons. Therefore, these results suggest a re-interpretation of several prior studies in comparative biology. We hope that these results contribute to the growing database of medaka fish phenotypes and provide one of the foundations for future phenomics studies of medaka fish.

A new species of Dixonius (Squamata: Gekkonidae) from Vientiane Capital, Laos

A new species of the gekkonid genus Dixonius is described from Huaysorn-Huaysua village, Nasaithong District, Vientiane Capital, Laos. The new species can be distinguished from all other species by a combination of the following characters: maximum SVL 47.1 mm; 19–21 longitudinal rows of dorsal tubercles at midbody; 23–26 longitudinal rows of ventrals across the abdomen; 7 or 8 supralabials, 6 in mid-orbital position; 5 or 6 infralabials; 5 or 6 precloacal pores in males, femoral pores lacking; precloacal and femoral pores absent in females; a canthal stripe running from rostrum through the eye and terminating at back of head; dorsum brown with irregular dark marks. In phylogenetic analyses, the species is recovered as the sister taxon to D. siamensis, which occurs in Laos, Thailand, and Vietnam. Genetically, the two species are approximately 9.4% divergent from each other based on the complete mitochondrial ND2 gene with six partial or complete adjacent tRNAs. It is the 12th species known in the genus Dixonius and the second Dixonius species described from Laos.

Genetic Identification of Edible Bird’s Nest in Thailand Based on ARMS-PCR

Edible bird’s nest (EBN) is a popular delicacy in the Asian Pacific region originating from Indonesia, Malaysia, Thailand and Vietnam, which consist of various potential medicine value in Traditional Chinese Medicine (TCM). Thailand is one of the main exporters of EBN. However, the genetic information of EBN, a key part of molecular biology, has yet to be reported in Thailand. It is necessary to explore the genetic information of EBN in Thailand based on a quick and simple method to help protect the rights and interests of consumers. This research aimed to systematically evaluate different methods of extracting EBN DNA to improve the efficiency of the analysis of cytochrome b (Cytb) and NADH dehydrogenase subunit 2 (ND2) gene sequences, the establishment of phylogenetic trees, and the genetic information of EBN in Thailand. Additionally, we aimed to develop a quick and simple method for identifying EBN from different species based on the genetic information and amplification-refractory mutation system PCR (ARMS-PCR). By comparing the four methods [cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS), kit and guanidinium isothiocyanate methods] for EBN extraction, we found that the guanidinium isothiocyanate method was the optimal extraction method. Phylogenetic trees generated on the basis of Cytb and ND2 gene analyses showed that 26 samples of house EBN and 4 samples of cave EBN came from Aerodramus fuciphagus and Aerodramus maximus, respectively. In addition, to distinguish different samples from different species of Apodiformes, we designed 4 polymerase chain reaction (PCR) amplification primers based on the ND2 gene sequences of A. fuciphagus and A. maximus. The ARMS-PCR results showed band lengths for A. fuciphagus EBN of 533, 402, and 201 bp, while those for A. maximus EBN were 463, 317, and 201 bp. Collectively, the results showed that ARMS-PCR is a fast and simple method for the genetic identification of EBN based on designing specific original identification primers.

Two new species of Hemiphyllodactylus (Squamata: Gekkonidae) from Hoa Binh Province, Vietnam

We describe two new species of the genus Hemiphyllodactylus on the basis of a new collection of geckos from limestone karst forest of Hoa Binh Province, northwestern Vietnam. Hemiphyllodactylus bonkowskii sp. nov. from Hang Kia—Pa Co Nature Reserve and Hemiphyllodactylus ngocsonensis sp. nov. from Ngoc Son—Ngo Luong Nature Reserve can be distinguished from each other and from their congeners by genetic distinction and morphological differences in circumnasal scales, chin scales, precloacal and femoral pores, cloacal spurs, dorsal scales, ventral scales, digital lamellae formula, and color pattern. In phylogenetic analyses, both new species are nested in a clade that includes H. dushanensis, H. flaviventris, H. hongkongensis, H. huishuiensis, H. kiziriani, H. serpispecus, and H. zugi. In terms of pairwise genetic distance, the two new species are at least 7.5%–8.5% divergent from other described species based on a fragment of the ND2 gene.

First Records of Bufo luchunnicus (Yang et Rao, 2008) and Amolops wenshanensis Yuan, Jin, Li, Stuart et Wu, 2018 (Anura: Bufonidae, Ranidae) from Vietnam

We record two species of amphibians for the first time from Vietnam: Bufo luchunnicus from Lao Cai and Son La provinces and Amolops wenshanensis from Quang Ninh Province. Morphologically, the Vietnamese representatives of B. luchunnicus resemble the type series from China. The specimen of A. wenshanensis from Vietnam slightly differs from the type series from China by having a smaller size (SVL 33.2 mm vs. 35.7 – 39.9 mm in males) and the presence of distinct transverse bands on the dorsal surfaces of limbs. Genetic divergence between the sequence of the Vietnamese specimen and those of A. wenshanensis from China available from GenBank is 1.2 – 1.6% (ND2 gene). In addition, morphological data and natural history notes of aforementioned species are provided from Vietnam.