A new species of the genus Ptyctolaemus Peters, 1864 (Squamata, Agamidae) from Sagaing, Myanmar

A new species of Ptyctolaemus Peters, 1864 is described from Htamanthi Wildlife Sanctuary, Sagaing Division, Myanmar. The new species differs from P. gularis and Ptyctolaemus aff. gularis from Tibet, China, by having relatively longer limbs and different colorations of the gular region, and it differs from P. collicristatus by having much longer limbs and a less developed nuchal crest in males. Moreover, the new species differs genetically from Ptyctolaemus aff. gularis from Tibet, China, and P. collicristatus by an uncorrected percentage distance of 23.5% and 24.8%, respectively, inferred from mitochondrial NADH dehydrogenase subunit 2 gene sequences. This discovery increases the number of known Ptyctolaemus species to three.

A new cryptic arboreal species of the Cyrtodactylus brevipalmatus group (Squamata: Gekkonidae) from the uplands of western Thailand

The first integrative taxonomic analysis of the Cyrtodactylus brevipalmatus group of Southeast Asia recovered two newly discovered populations from the Tenasserim Mountains in Suan Phueng District, Ratchaburi Province, Thailand as a new species described here as C. rukhadeva sp. nov. Based on 1397 base pairs of the mitochondrial gene NADH dehydrogenase subunit 2 (ND2), C. rukhadeva sp. nov. is the well-supported sister species to a clade containing three undescribed species, C. ngati, and C. cf. interdigitalis with a large uncorrected pairwise sequence divergence from other species in the brevipalmatus group ranging from 15.4–22.1%. Cyrtodactylus elok and C. brevipalmatus are recovered as poorly supported sister species and the well-supported sister lineage to the remainder of the brevipalmatus group. Cyrtodactylus rukhadeva sp. nov. is putatively diagnosable on the basis of a number of meristic characters and easily separated from the remaining species of the brevipalmatus group by a number of discrete morphological characters as well as its statistically significant wide separation in multivariate morphospace. The discovery of C. rukhadeva sp. nov. continues to underscore the unrealized herpetological diversity in the upland forests of the Tenasserim Mountains and that additional field work will undoubtedly result in the discovery of additional new species.

A new species of the genus Tylototriton (Urodela, Salamandridae) from western Thailand

We describe a new species of the newt genus Tylototriton from Umphang Wildlife Sanctuary, Tak Province, western Thailand based on molecular and morphological evidence and named here as Tylototriton umphangensissp. nov. The new species is assigned to the subgenus Tylototriton and differs from other species in having dark-brown to blackish-brown body and limbs, truncate snout, prominent antero-medial ends of the expansion of the dentary bones, laterally protruding quadrate regions, indistinct and small rib nodules, a well-segmented vertebral ridge, and rough dorsolateral bony ridges, which are steeper anterior, and curved medially at the posterior ends. The molecular data show that Tylototriton umphangensissp. nov. differs from T. uyenoi sensu stricto by a 5% genetic sequence divergence of the mitochondrial NADH dehydrogenase subunit 2 region gene. The new species and T. uyenoi are both endemic to Thailand, distributed along the Northwest Thai (Dawna) Uplands of Indochina. To clarify the species boundary between Tylototriton umphangensissp. nov. and T. uyenoi, additional field research is needed in adjacent areas. Tylototriton umphangensissp. nov. is restricted to evergreen hill forests in Umphang Wildlife Sanctuary. We suggest that the new species should be classified as Endangered (EN) in the IUCN Red List.

A case of mistaken identity: Genetic and morphological evidence for the presence of Redeye Bass in the Verde River, Arizona

We report genetic and morphological evidence for the presence of Redeye Bass Micropterus coosae, in the Verde River of Arizona, previously thought to be Smallmouth Bass Micropterus dolomieu. We performed meristic measurements on 15 individuals sampled from the Upper Verde River Wildlife Area, Yavapai County, Arizona. Meristic data for lateral line scales, scales above lateral line, and scales below lateral line were all consistent with Redeye Bass and not Smallmouth Bass. We analyzed mitochondrial and nuclear genetic data to determine if one of the black bass (Genus Micropterus) species historically introduced to the Verde River was Redeye Bass and that they persist in the system. We extracted DNA from fin clips of five individuals for phylogenetic analysis of the NADH dehydrogenase subunit 2 (ND2) mitochondrial gene and for analysis of nuclear DNA using a diagnostic Single Nucleotide Polymorphism (SNP) panel. Results of the ND2 genetic sequencing and phylogenetic analysis indicated that these fish likely originated from native Redeye Bass stock from the Coosa River system of Alabama, Georgia, and Tennessee. Similarly, nuclear SNP data from the five individuals collected from the Verde River aligned with Redeye Bass reference genotypes based on STRUCTURE analysis. These results support the hypothesis that at least one of the introductions of black bass in Arizona’s Verde River founded a previously unrecognized population of Redeye Bass. Further work is needed to determine the extent of the Redeye Bass presence in Arizona, whether Smallmouth Bass are also present in the Verde River system, and if hybridization of Redeye Bass and other black basses is occurring.

Brown banded bamboo shark (Chiloscyllium punctatum) shows high genetic diversity and differentiation in Malaysian waters

The demersal brown banded bamboo shark Chiloscyllium punctatum is a major component of sharks landed in Malaysia. However, little is known about their population structure and the effect of high fishing pressure on these weak swimming sharks. Both mitochondrial DNA control region (1072 bp) and NADH dehydrogenase subunit 2 (1044 bp) were used to elucidate the genetic structure and connectivity of C. punctatum among five major areas within the Sundaland region. Our findings revealed (i) strong genetic structure with little present day mixing between the major areas, (ii) high intra-population genetic diversity with unique haplotypes, (iii) significant correlation between genetic differentiation and geographical distance coupled with detectable presence of fine scale geographical barriers (i.e. the South China Sea), (iv) historical directional gene flow from the east coast of Peninsular Malaysia towards the west coast and Borneo, and (v) no detectable genetic differentiation along the coastline of east Peninsular Malaysia. Genetic patterns inferred from the mitochondrial DNA loci were consistent with the strong coastal shelf association in this species, the presence of contemporary barriers shaped by benthic features, and limited current-driven egg dispersal. Fine scale population structure of C. punctatum highlights the need to improve genetic understanding for fishery management and conservation of other small-sized sharks.

Protective Effect of Mitochondrial ND2 C5178A Gene Mutation on Cell and Mitochondrial Functions

Background. Mitochondrial NADH dehydrogenase subunit 2 (MT-ND2) m. 5178C>A gene mutation has protective effects against various diseases, but the molecular mechanism is still unclear. In previous study, we found a heteroplasmy level of MT-ND2 m. 5178C>A mutation in normotensive controls. Peripheral blood samples were obtained from essential hypertension individuals carrying the mutation and healthy controls without gene mutation to establish immortalized lymphocyte lines. To investigate the effect of the MT-ND2 m. 5178C>A gene mutation, comparative analyses of the two group cell lines were performed, including measurements of cell proliferation, viability, ATP synthesis, mitochondrial oxidative stress, and oxidative phosphorylation. Results. The cell proliferation rate and viability of the MT-ND2 m. 5178C>A mutant lymphocyte line were higher than those of the control group. Mitochondrial functions of the MT-ND2 m. 5178C>A mutant lymphocyte were increased, including increased ATP synthesis, decreased ROS production, increased mitochondrial membrane potential and Bcl-2 gene transcription and protein translation, decreased Caspase 3/7 activity, and decreased early apoptosis and late apoptosis. The oxygen consumption rate (OCR) of the mutant lymphocyte line was higher than that of the control group, including basal OCR, ATP-linked OCR, maximal OCR, proton leak OCR, and reserve OCR, and there was no significant difference in nonmitochondrial OCR. The activity of Mitochondrial Complex I of the mutant group was increased than that of the control group. Conclusions. The MT-ND2 m. 5178C>A mutation is a protective mutation that may be related to improvement of mitochondrial functions and decrease in apoptosis.

Onchocerca Flexuosa. sp. (Nematoda: Filarioidea) in Japanese Wild Sika Deer (Cervus Nippon): Pathological and Molecular Identification of Intradermal Worms

More than thirty species of the genus Onchocerca (Nematoda; Filarioidea) were responsible for onchocercosis (a vector-borne parasitic disease) of wild and domestic ungulates. “Between 2016 and 2017”, the presence of Onchocerca flexuosa (Wedl, 1856) was detected and investigated in 17 Japanese Sika deer (Cervus nippon) captured in Gifu and Shiga Prefectures, Japan. The worms were found within characteristics subcutaneous nodules dispersed in different regions of the body of all 17 deer. The all collected nodules were examined stereo-microscopically. The parasites were extracted from the nodules and identified through morphological and histopathological examinations. Molecular identification through sequencing of the following genes; internal transcribed spacer subunit 2 (ITS2)–28S ribosomal RNA (28S rRNA), cytochrome c oxidase subunit 1 (cox1) and mitochondrially encoded NADH dehydrogenase subunit 2 (NAD2) were performed. The histopathological, molecular and phylogenetic analysis demonstrated that, the filarial nematode isolated from Gifu and Shiga Prefectures in Japan is O. flexuosa. This is the first report about presence of O. flexuosa in Japanese Sika deer (Cervus nippon) in Gifu and Shiga Prefectures.

A new species of Hemiphyllodactylus Bleeker (Squamata: Gekkonidae) from Yunnan, China and its phylogenetic relationship to other congeners

A new species of the gekkonid genus Hemiphyllodactylus is described from forested karst hills near Zhutangxiang town, Lancang Lahu Autonomous County, Yunnan, China. Hemiphyllodactylus zhutangxiangensis sp. nov. is distinguished from all other congeners in morphology and an uncorrected pairwise sequence divergence of greater than 14% based on 1039 base pairs of the mitochondrial NADH dehydrogenase subunit 2 gene (ND2). The new species is defined by the following characters: a maximum SVL of 44.42 mm; 7–9 chin scales; enlarged postmentals; five circumnasal scales; 2–4 internasal scales; 8–11 supralabial scales; 8–11 infralabial scales; subdigital lamellae on fingers II–V (3–5)-(4–6)-(4 or 5)-(4 or 5); subdigital lamellae on toes II–V (4 or 5)-(4 or 5)-(4–6)-(4 or 5); dorsal scales 11–15; ventral scales 5–7; 20–23 continuous femoroprecloacal pores; having pale-grey base color on the body with various darker transverse dorsal blotches; a dark postorbital stripe extending to at least to the base of the neck; no dark dorsolateral or ventrolateral stripe on trunk; and postsacral marking bearing a dark fork-like pattern with anteriorly projecting arms. The new species brings the species total of Hemiphyllodactylus in China to 11.

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.

Molecular phylogenetics, PCA, and MFA recover a new species of Cyrtodactylus (Squamata: Gekkonidae) from an isolated sandstone massif in northwestern Cambodia

The integrated results of maximum likelihood (ML) and Bayesian inference (BI) analyses, principal component analyses (PCA), and a multiple factor analysis (MFA) recover a new, widely allopatric species of the Cyrtodactylus intermedius species group. Cyrtodactylus kulenensis sp. nov is endemic to the Phnom Kulen sandstone massif of the Phnom Kulen National Park, Siem Reap Province, in the lowlands of northwestern Cambodia. A phylogenetic analysis from a short read (275 base pairs) of the mitochondrial gene NADH dehydrogenase subunit 2 (ND2) from C. kulenensis sp. nov. was aligned with 1449 base pairs from all other species in the intermedius group. The analysis recovered C. kulenensis sp. nov. as the sister species to a lineage composed of populations from the widely separated hilly regions of Sa Keao and Sakaerat in eastern Thailand. Multivariate (PCA, DAPC, and MFA) and univariate analyses (ANOVA) using combinations of meristic (scale counts), mensural (morphometric), and categorical (color pattern and morphology) characters from 52 specimens encompassing all species of the intermedius group clearly demonstrate C. kulenensis sp. nov. is significantly different and discretely diagnosable from all other species in the intermedius group. This new discovery further highlights the herpetological diversity and high levels of range-restricted endemism in basin-habitat-island landscapes throughout Indochina and the continued need for field work in the landscapes that remain unsurveyed.