Phylogenetic relationships and genetic diversity of the Polypedates leucomystax complex in Thailand

Taxonomic uncertainty of the Asian tree frog Polypedates leucomystax complex presents the challenging task of inferring its biogeographical history. Here, we describe its dispersion and the genetic relationships among different populations in Thailand, where we connect the population of the P. leucomystax complex of the Sunda Islands to the Indochina (mainland) population based on analyses of 266 sequences of the mitochondrial cytochrome c oxidase subunit I (COI) gene. Our maternal genealogy implies that there are four well-supported lineages in Thailand, consisting of Northern A (clade A: Polypedates sp.), Nan (clade B: P. cf. impresus), Southern (clade C: P. cf. leucomystax) and Northern D (clade D: P. cf. megacephalus), with Bayesian posterior probability >0.9. Phylogeny and haplotype networks indicate that clades A, B and D are sympatric. In contrast, clade C (P. cf. leucomystax) and clade D (P. cf. megacephalus) are genetically divergent due to the geographical barrier of the Isthmus of Kra, resulting in an allopatric distribution. Climatic conditions, in particular differences in rainfall on each side of the Isthmus of Kra, may play an important role in limiting the immigration of both clades. For the within-populations of either clades C or D, there was no significant correlation between geographic and genetic distance by the isolation-by-distance test, indicating intraspecific-dispersal of each clade. Population expansion occurred in clade C, whereas clade D showed a constant population. Taken together, the P. leucomystax complex in South East Asia may have diversified under climatic pressure, leading to allopatric and/or sympatric speciation

In vitro chondroprotective potential of Senna alata and Senna tora in porcine cartilage explants and their species differentiation by DNA barcoding-high resolution melting (Bar-HRM) analysis.

Senna species and anthraquinone derivatives generated by these organisms, rhein and aloe-emodin, exert anti-inflammatory effects. These species present a similar morphology but produce different ingredients when they are used as medicinal products. In this study, a DNA barcoding- (Bar-) high-resolution melting (HRM) technique was developed using internal transcribed sequence 2 (ITS2) to differentiate between Senna alata and Senna tora as a result of significant differences in their melting profiles. We used this approach for confirmation of S. alata and S. tora raw materials, and we examined the chondroprotective properties of the ethanolic extracts of S. alata and S. tora using a porcine model of cartilage degradation induced by a combination of interleukin-17A (IL-17A) and IL-1β. We found that both Senna ethanolic extracts, at a concentration of 25 μg/mL, effectively prevented cartilage degradation. Rhein and aloe-emodin were present in the extract of S. alata but not in that of S. tora. We observed a reduction in the release of sulfated glycosaminoglycans (S-GAGs) and hyaluronic acid (HA) into media in both treatments of Senna extracts, which indicated proteoglycan preservation in explant tissues. These results suggest that neither rhein nor aloe-emodin are the main factors responsible for cartilage-protecting properties. Taken together, results show that both S. alata and S. tora are promising for further development as anti-osteoarthritic agents and that Bar-HRM using ITS2 could be applied for species confirmation with Senna products

Post-treatment of hyaluronan to decrease the apoptotic effects of carprofen in canine articular chondrocyte culture

A major concern associated with the use of drugs is their adverse side effects. Specific examples of the drugs of concern include antibiotic agents and non-steroidal anti-inflammatory drugs. Despite the presence of a high degree of efficacy for specific conditions, these drugs may deteriorate the surrounding tissues that are exposed to them. Often, carprofen is used for joint inflammation; however, it may stimulate cartilage degradation which can then lead to osteoarthritis progression. In this study, hyaluronan was combined with carprofen treatment in three different applications (pre-treatment, co-treatment and post-treatment) on normal canine chondrocytes to determine whether Hyaluronan (HA) is capable of mitigating the degree of chondrotoxicity of carprofen. Our findings revealed that carprofen at IC20 (0.16 mg/mL) decreased viability and increased nitric oxide (NO) production. Importantly, carprofen induced the apoptosis of canine chondrocytes via the up-regulation of Bax, Casp3, Casp8, Casp9 and NOS2 as compared to the control group. Although the co-treatment of HA and carprofen appeared not to further alleviate the chondrotoxicity of carprofen due to the presence of a high number of apoptotic chondrocytes, post-treatment with HA (carprofen treatment for 24 h and then changed to HA for 24 h) resulted in a decrease in chondrocyte apoptosis by the down-regulation of Bax, Casp3, Casp8, Casp9, NOS2, along with NO production when compared with the treatment of carprofen for 48 h (P < 0.05). These results suggest that HA can be used as a therapeutic agent to mitigate the degree of chondrotoxicity of carprofen

Dystroglycan 1: A new candidate gene for patellar luxation in Chihuahua dogs

Aim: The objective of this study was to uncover new candidate genes related to patellar luxation (PL) in dogs to select for those with low susceptibility for breeding purposes. Materials and Methods: The inter simple sequence repeat (ISSR) technique was performed to construct DNA fingerprints of 61 Chihuahua dogs with PL and 30 healthy Chihuahua dogs. DNA polymorphisms were detected by comparing the sequences between the affected and unaffected dogs, using the pairwise alignments in MultAlin. Genotyping was performed using allele-specific polymerase chain reaction (AS-PCR). The association analysis of ISSR DNA fingerprints and genotypes or phenotypes was performed using the Chi-square (χ2) model and generalized linear model (GLM), respectively. Results: Two single nucleotide polymorphisms (SNPs), namely SNP1UBC811 (g.91175C>G) and SNP2UBC811 (g.92259T>C), were found in the intron of the Dystroglycan 1 (DAG1) gene, which was obtained using the PL-related marker UBC811 primer (p=0.02), and genotyped by AS-PCR. When investigated using the GLM, g.91175C>G had a significant association with PL (p=0.0424), whereas g.92259T>C did not have such an association (p=0.0959). Conclusion: DAG1 might be one of the genes related to PL in Chihuahuas and could aid the process of marker-assisted selection in genetic breeding for Chihuahua dogs without PL

The Potential Distribution and Maternal Lineage of Two Cetaceans Species (Grampus griseus and Pseudorca crassidens) in the Subfamily Globicephalinae from the Thai Andaman Sea and the Gulf of Thailand

Although the existence of two cetacean species in and around Thai Seas from within the subfamily Globicephalinae, Risso&rsquo;s dolphin Grampus griseus G. Cuvier, 1812, and false killer whale Pseudorca crassidens Owen, 1846, has been known for decades, current knowledge on the abundance, genetic diversity, and conservation status of these marine mammals is limited as these are rare oceanic species for Thailand&rsquo;s territorial waters. Frozen skin tissue samples taken from six cetaceans (four Risso&rsquo;s dolphins: two false killer whales) stranded along Thai coastlines were investigated. We aimed to identify the maternal lineage and connection of our samples throughout their distribution range. Accordingly, we analyzed the dataset of 110 and 50 mtDNA control region sequences of Risso&rsquo;s dolphins and false killer whales, respectively. This dataset was retrieved from the online database of the National Center for Biotechnology Information (NCBI) and included six mtDNA sequences obtained from Thai Seas. Two unique haplotypes of Risso&rsquo;s dolphins were found in the Thai Andaman Sea, whereas one haplotype identified as being from the Gulf of Thailand is a common haplotype shared with other regions of the Pacific Ocean. Two haplotypes were found for false killer whales from the Thai Andaman Sea, and these were also in common with other regions of the Indo Pacific Ocean. While shared haplotypes with other regions may imply inheritance from the same female ancestor, we speculate that distinct populations with unique genetic structures also exist in Thai Seas. Beneficially, our results could be used to monitor alterations of haplotypes or to assess the maternal genetic diversity of designated species in the future to establish baseline information for Thai Seas and adjacent waters

Genetic Diversity in Leatherback Turtles (Dermochelys coriacea) along the Andaman Sea of Thailand

The leatherback sea turtle (Dermochelys coriacea) is the largest and one of the most migratory turtle species, inhabiting oceans throughout the world. There has been a steady decline in leatherback populations over the past several decades due to human activities. They are considered endangered in Thailand and global, so conservation strategies are needed to study and protect the species, including determining their genetic diversity. A total of 8 microsatellite loci and 658 bp amplicon of mitochondrial DNA (mtDNA) were used to assess genetic data from 149 dead leatherback turtle hatchlings among 14 nests in five locations along the Andaman Sea, Thailand, between 2018&ndash;2020. The microsatellite findings show that the observed heterozygosity (Ho) ranged from 0.44 &plusmn; 0.09 to 0.65 &plusmn; 0.10. Population structures were further divided into two genetically distinct groups by Bayesian inference. For the mtDNA control region, our samples consisted of three haplotypes. Globally, there are 27 haplotypes of leatherback turtles, with a relatively low genetic diversity (h = 0.43). These results reveal the genetic status of leatherback turtles in Thailand and globally, and raise concerns about their relative genetic health, which highlight the need for proactive, long-term management and conservation strategies for this endangered species

Genetic Diversity and Maternal Lineage of Indo-Pacific Bottlenose Dolphin (<i>Tursiops aduncus</i>) in the Andaman Sea of Thailand

Indo-Pacific bottlenose dolphins (Tursiops aduncus) are a coastal species found in Thai waters off the coasts of the Andaman Sea and the Gulf of Thailand. This species was recently re-listed as near-threatened by the IUCN Red List, though the population status in Thai seas is not known. Here, we investigated genetic diversity, population structure, maternal lineage, and demographics by analyzing skin tissue samples (n = 30) of T. aduncus stranded along the Andaman coastline of Thailand between 1990 and 2019. This study was based on 11 microsatellite loci and 265 bp mtDNA control regions compared to data available through the National Center for Biotechnology Information (NCBI). From microsatellites, the observed heterozygosity (Ho) ranged from 0.46 to 0.85. The mean fixation index (F) value for all loci was 0.10 ± 0.04, which suggests some degree of inbreeding. Two genetic clusters (the most likely K at K = 2) were observed in T. aduncus through the population structure analysis using multiple criteria. For the mtDNA control region, a total of 17 haplotypes were found for dolphins in Thai seas (14 haplotypes from our samples; three haplotypes from the NCBI database) with high levels of haplotype diversity (h) at 0.926 ± 0.027 and nucleotide diversity (π) at 0.045 ± 0.002. A decline in the effective population size from 0.05 million years ago also was observed in Thai T. aduncus through Bayesian Skyline Plots analysis. A unique set of haplotypes was identified in our samples, which may have originated from the Australian and Indian Oceans rather than the Western Pacific Ocean. These results improve our understanding of the maternal lineage of the Indo-Pacific bottlenose dolphin, which can be used for monitoring population status and establishing better conservation plans for this species in the Thai Andaman Sea