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Centella asiatica (L.) Urb. has wound healing, anti-inflammatory, cognitive improvement, and neuroprotective properties which have been attributed to its centelloside content. However, the quantities of these bioactive compounds are limited and vary due to genetic and environmental factors. Light qualities are known to enhance the production of secondary metabolites in several plant species, both preharvest and postharvest. In this study, fresh leaves of C. asiatica were subjected to different light emitting diode (LED) quality including white, dark, red, blue, and green to assess centelloside content, phytochemical constituents, and transcription level expression of triterpenoid biosynthesis genes. Results showed that white and blue LEDs significantly increased centelloside content in C. asiatica leaves at 3 days postharvest (dph) by 73 % over the control group at 0 dph. Blue LEDs stimulated the expression of triterpenoid biosynthesis genes including C. asiatica squalene synthase (CaSQS), C. asiatica β-amyrin synthase (CabAS), and C. asiatica UDP gluclosyltransferase-73AH1 (CaUGT73AH1; CaUGT), while different LED conditions gave diverse results. Red LED treatment triggered higher total flavonoid content (TFC) and total triterpenoid content (TTC) while white LEDs enhanced total triterpenoid content (TTC). Taken together, our findings suggest that postharvest under blue LEDs is a great approach to increase centelloside production of C. asiatica through gene up-regulation in triterpenoid pathway. Therefore, postharvest technology by LEDs serves as an effective tool for improving raw material quality for medicinal plant industries.

Magnetoreception, the remarkable ability of organisms to perceive and respond to Earth’s magnetic field, has captivated scientists for decades, particularly within the field of quantum biology. In the plant science, the exploration of the complicated interplay between quantum phenomena and classical biology in the context of plant magnetoreception has emerged as an attractive area of research. This comprehensive review investigates into three prominent theoretical models: the Radical Pair Mechanism (RPM), the Level Crossing Mechanism (LCM), and the Magnetite-based MagR theory in plants. While examining the advantages, limitations, and challenges associated with each model, this review places a particular weight on the RPM, highlighting its well-established role of cryptochromes and in-vivo experiments on light-independent plant magnetoreception. However, alternative mechanisms such as the LCM and the MagR theory are objectively presented as convincing perspectives that permit further investigation. To shed light on these theoretical frameworks, this review proposes experimental approaches including cutting-edge experimental techniques. By integrating these approaches, a comprehensive understanding of the complex mechanisms driving plant magnetoreception can be achieved, lending support to the fundamental principle in the RPM. In conclusion, this review provides a panoramic overview of plant magnetoreception, highlighting the exciting potential of quantum biology in unraveling the mysteries of magnetoreception. As researchers embark on this captivating scientific journey, the doors to deciphering the diverse mechanisms of magnetoreception in plants stand wide open, offering a profound exploration of nature’s adaptations to environmental cues.

Abstract Canine babesiosis is a tick-borne disease caused by Babesia spp., which infects and destroys healthy erythrocytes, leading to mortality and morbidity in dogs. The diagnosis of babesiosis is tedious and time-consuming, especially in latent and chronic infections. Here, a recombinase polymerase amplification combined with a lateral flow dipstick (RPA-LFD) assay was developed for rapid and accurate detection of Babesia spp. in canine blood specimens based on the 18S rRNA region. The RPA-LFD assay using rpaBab264 gave specificity to Babesia spp. in dogs (B. vogeli and B. gibsoni) without cross-amplification to other parasites (apicomplexans and non-apicomplexans), with detection limit of at least 22.5 copies/μl (0.1 fg/µl) at 40 °C for at least 10 min. The whole process of DNA amplification by RPA and readout by LFD did not exceed 30 min. To determine the performance of the RPA-LFD assay, a total of 30 clinical samples was examined and compared with conventional PCR (cPCR) and multiplex HRM (mHRM). Eight dogs (26.67%) were detected as positive by RPA-LFD, while seven and six were found positive by cPCR and mHRM, respectively. RPA-LFD and cPCR showed high agreement with Babesia spp. detection with kappa > 0.9. We confirmed that the dogs were infected by B. vogeli from sequences of positive PCR results. Our findings suggested that RPA-LFD using the rpaBab264 assay offered a rapid, accurate, cost-effective and simple method for Babesia spp. detection that is feasibly applicable to be rapid kit at a pet hospital or point-of-care testing.

Background and Aim: The Indo-Pacific bottlenose dolphin, Tursiops aduncus, and the pantropical spotted dolphin, Stenella attenuata, are protected marine mammals in Thailand; however, knowledge regarding the populations of both species in Thai seas is minimal. We aimed to reveal the genetic diversity and population structure of two species, T. aduncus, and S. attenuata, based on inter-simple sequence repeats (ISSRs). Materials and Methods: Samples of stranded T. aduncus (n = 30) and S. attenuata (n = 23) found along Thai Andaman Sea coasts from 1998 to 2018 were used in this study. A total of 17 and 16 ISSR primers that produced clear and polymorphic bands were selected for T. aduncus and S. attenuata, respectively. Results: The highest percentages of polymorphic bands for T. aduncus and S. attenuata were 93.750% and 92.857%, respectively. Phylogenetic dendrograms indicated that the population of each species was clustered into three groups. This outcome was consistent with the genetic population structure, as both suggested three genetic clusters (ΔK = 3). Genetic diversity analysis revealed that the average Shannon's information index (I) was 1.926 ± 0.066 for T. aduncus and 1.714 ± 0.090 for S. attenuata, which indicate a high level of genetic variation. Further, low fixation index (F) values were observed for T. aduncus and S. attenuata at –0.231 ± 0.024 and –0.312 ± 0.042, respectively, suggesting that inbreeding is unlikely to have occurred for both species over the past decades. Conclusion: At least three genetic clusters of both species were found in the Thai Andaman Sea, and the diversity indices of each species indicated that these species are not at a critical level for extinction. However, monitoring their population status should be prioritized to observe any future changes in the level of diversity.

Xanthomonas oryzae pv. oryzae (Xoo) is a widespread pathogen causing bacterial leaf blight (BLB) disease, devastating rice productivity in many cultivated areas of Thailand. A specific and simple method for Xoo detection is required to improve surveillance of disease transmission and outbreak. This study developed a recombinase polymerase amplification (RPA) assay assisted with CRISPR-cas12a assay (RAC) for Xoo detection from bacterial cell suspension of infected rice samples without DNA extraction. The efficiency of the RAC system for Xoo detection using either Xoo80 or Xoo4009 locus was optimized to amplify and determine the sensitivity and specificity using a Xoo DNA template from bacterial cell suspension of infected rice samples without DNA extraction. The RAC system using the Xoo4009 locus gave a higher specificity than Xoo80 locus, because only Xoo species was amplified positive RPA product with fluorescence signal by cas12a digestion, which indicated no cross reactivity. Optimal RAC using the Xoo4009 locus enabled diagnosis of Xoo presence from both plant extracted samples of Xoo artificially inoculated rice leaves within 3 d post-inoculation without symptomatic BLB appearance, and Xoo naturally infected rice. Findings exhibited that RAC using the Xoo4009 locus offered sensitivity, specificity and simplicity for Xoo detection, with low intensities of Xoo-DNA (1 × 103 copies/μL) and Xoo-cell (2.5 × 103 cfu/mL). This developed RAC system showed significantly potential for Xoo detection at point-of-care application for early signs of BLB disease outbreak in rice fields.

Abstract Rapid and accurate species diagnosis accelerates performance in numerous biological fields and associated areas. However, morphology-based species taxonomy/identification might hinder study and lead to ambiguous results. DNA barcodes (Bar) has been employed extensively for plant species identification. Recently, CRISPR-cas system can be applied for diagnostic tool to detect pathogen’s DNA based on the collateral activity of cas12a or cas13. Here, we developed barcode-coupled with cas12a assay, “Bar-cas12a” for species authentication using Phyllanthus amarus as a model. The gRNAs were designed from trnL region, namely gRNA-A and gRNA-B. As a result, gRNA-A was highly specific to P. amarus amplified by RPA in contrast to gRNA-B even in contaminated condition. Apart from the large variation of gRNA-A binding in DNA target, cas12a- specific PAM’s gRNA-A as TTTN can be found only in P. amarus. PAM site may be recognized one of the potential regions for increasing specificity to authenticate species. In addition, the sensitivity of Bar-cas12a using both gRNAs gave the same detection limit at 0.8 fg and it was 1,000 times more sensitive compared to agarose gel electrophoresis. This approach displayed the accuracy degree of 90% for species authentication. Overall, Bar-cas12a using trnL-designed gRNA offer a highly specific, sensitive, speed, and simple approach for plant species authentication. Therefore, the current method serves as a promising tool for species determination which is likely to be implemented for onsite testing.

Abstract Dugong (Dugong dugon) populations have been shrinking globally, due in large part to habitat fragmentation, degradation and ocean pollution, and today are listed as Vulnerable by the IUCN. Thus, determining genetic diversity in the remaining populations is essential for conservation planning and protection. In this study, measures of inter-simple sequence repeat (ISSR) markers and mtDNA D-loop typing were used to evaluate the genetic diversity of 118 dugongs from skin samples of deceased dugongs collected in Thai waters over a 29-year period. Thirteen ISSR primers revealed that dugongs from the Andaman Sea and Gulf of Thailand exhibited more genetic variation in the first 12 years of the study (1990–2002) compared to the last decade (2009–2019). Dugongs from the Andaman Sea, Trang, Satun and some areas of Krabi province exhibited greater diversity compared to other coastal regions of Thailand. Eleven haplotypes were identified, and when compared to other parts of the world (235 sequences obtained from NCBI), five clades were apparent from a total 353 sequences. Moreover, dugongs from the Andaman Sea were genetically distinct, with a separate haplotype belonging to two clades found only in Thai waters that separated from other groups around 1.2 million years ago. Genetic diversity of dugongs in present times was less than that of past decades, likely due to increased population fragmentation. Because dugongs are difficult to keep and breed in captivity, improved in situ conservation actions are needed to sustain genetically healthy wild populations, and in particular, the specific genetic group found only in the Andaman Sea.

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.

Marine mammals vary greatly in size and lifespan across species. This study determined whether measures of adult body weight, length and relative telomere length were related to lifespan. Skin tissue samples (n = 338) were obtained from 23 marine mammal species, including four Mysticeti, 19 Odontoceti and one dugong species, and the DNA extracted to measure relative telomere length using real-time PCR. Life span, adult body weight, and adult body length of each species were retrieved from existing databases. The phylogenetic signal analysis revealed that body length might be a significant factor for shaping evolutionary processes of cetacean species through time, especially for genus Balaenoptera that have an enormous size. Further, our study found correlations between lifespan and adult body weight (R2 = 0.6465, p < 0.001) and adult body length (R2 = 0.6142, p ≤0.001), but no correlations with relative telomere length (R2 = −0.0476, p = 0.9826). While data support our hypothesis that larger marine mammals live longer, relative telomere length is not a good predictor of species longevity.

We established a method to test if bones have been varnished with lacquer. Handheld X-ray fluorescence was employed to detect the elemental at the surface of porcine femoral bones (n=10) that served as lacquer-free bones (control group). Eight elements – silicon (Si), phosphorus (P), sulfur (S), calcium (Ca), titanium (Ti), zinc (Zn), cadmium (Cd) and tin (Sn) – plus light elements (LE) showed a significant discrepancy between lacquer-varnished and lacquer-free bones (p

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 2018–2020. The microsatellite findings show that the observed heterozygosity (Ho) ranged from 0.44 ± 0.09 to 0.65 ± 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.

Currently, species identification of stranded marine mammals mostly relies on morphological features, which has inherent challenges. The use of genetic information for marine mammal species identification remains limited, therefore, new approaches that can contribute to a better monitoring of stranded species are needed. In that context, the ISSR-HRM method we have proposed offers a new approach for marine mammal species identification. Consequently, new approaches need to be developed to identify individuals at the species level. Eight primers of the ISSR markers were chosen for HRM analysis resulting in ranges of accuracy of 56.78–75.50% and 52.14–75.93% in terms of precision, while a degree of sensitivity of more than 80% was recorded when each single primer was used. The ISSR-HRM primer combinations revealed a success rate of 100% in terms of discrimination for all marine mammals included in this study. Furthermore, ISSR-HRM analysis was successfully employed in determining marine mammal discrimination among varying marine mammal species. Thus, ISSR-HRM analysis could serve as an effective alternative tool in the species identification process. This option would offer researchers a heightened level of convenience in terms of its performance and success rate. It would also offer field practice to veterinarians, biologists and other field-related people a greater degree of ease with which they could interpret results when effectively classifying stranded marine mammals. However, further studies with more samples and with a broader geographical scope will be required involving distinct populations to account for the high degree of intraspecific variability in cetaceans and to demonstrate the range of applications of this approach.

The ability to estimate age and determine the growth status of free-ranging dugongs (Dugong dugon) is vital to providing insight into the basic biology of this endangered species. Currently, age estimation in dugong carcasses relies on counting dentin growth layer groups (GLGs) in tusks, but a disadvantage is they need to be intact. We explored whether measures of telomere length could be used as an alternative approach to age estimation in dugongs given that in other species, telomere length and age are inversely related. In this study, relative telomere length (rTL) was measured by qPCR in skin samples from 24 dugongs of varying ages determined by counts of GLGs. In addition, relationships between age by GLG counts and body weight and length and were examined. Our findings indicate that age estimated by GLGs was negatively correlated with telomere length using the logistic formula with a rate of telomere attrition of approximately 0.036 rTL/year between the ages of 5–20 years. By comparison, both body weight and length were positively correlated with GLG-based age, with growth rates of ~8.8 kg/year for weight and ~3.58 cm/year for length, respectively. After that, growth rates slowed substantially and then plateaued. The results suggest that physical maturity in dugongs occurs at 20 years of age and that measures of rTL might serve as a tool for age estimation in dugongs, living and deceased.

The identification of differing physical characteristics of dogs is an uncomplicated and straightforward way to categorize dog breeds. However, many dog owners and veterinarians still struggle to distinguish between pure breed and mixed variations in certain breeds of dogs. Presently, the absence of the tools and methods needed to confirm a pure breed dog is a significant problem since the only method available to validate pure or mongrel breeds is the official pedigree system. Inter-simple sequence repeat markers have been successfully used to assess genetic variations and differentiations. Notably, inter-simple sequence repeat markers coupled with high resolution melting analysis were effectively used for the breed identification of 43 breeds of dogs (total 463 dogs). The 10 primers chosen for analysis resulted in a range of 31–78.6% of breed discrimination when using one primer, while a combination of two primers was able to successfully discriminate between all of the 43 dog breeds (100%). Shannon’s index information (I = 2.586 ± 0.034) and expected heterozygosity (He = 0.908 ± 0.003) indicated a high level of genetic diversity among breeds. The fixation index (Fst) revealed a value of 10.4%, demonstrating that there was a high level of genetic subdivision between populations. This study showed that inter-simple sequence repeat marker analysis was effective in demonstrating high genetic diversity among varying breeds of dogs, while a combination of Inter-simple sequence repeat marker analysis and high resolution melting analysis could provide an optional technique for researchers to effectively identify breeds through genetic variations.

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.

Although the existence of two cetacean species in and around Thai Seas from within the subfamily Globicephalinae, Risso’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’s territorial waters. Frozen skin tissue samples taken from six cetaceans (four Risso’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’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’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.

The dugong (Dugong dugon) is an endangered species of marine mammals, so knowledge of genetic diversity of these populations is important for conservation planning within different habitats. In this study, six microsatellite markers were used to assess the genetic diversity and population structure of 77 dugongs from skin samples of stranded animals collected from 1994–2019 (69 from Andaman Sea and 8 from the Gulf of Thailand). Our results found that dugongs in the Andaman Sea had higher genetic variation than those in the Gulf of Thailand. Populations in Trang, Satun, and some areas of Krabi had highest diversity compared to other regions of Thailand. Bayesian genetic clustering analysis revealed that dugongs in Thailand consist of five genetic groups. Moreover, dugongs in the middle and lower Andaman Sea presented the greatest gene flow compared to other regions. However, based on calculation of inbreeding coefficients (Fis value = 0.239), dugong populations in the Sea of Thailand are experiencing some levels of inbreeding, and so may warrant special protections. These results provide important information for understanding the genetic status of dugongs that can lead to improved management and conservation of this endangered species.

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.

This study conducted an investigation on three exons of the endothelin receptor type B (EDNRB) gene of Thai odd-eyed cats to find out the association between the variations in the gene and heterochromia iridis. DNA sequencing analysis was performed on 11 odd-eyed cats compared to 11 normal-eyed cats. Seven variations were found across the studied region (XM_003980457.2: c.610A>G, c.820+40C>T, c.821-14C>T, c.916A>G, c.1025+36G>T, c.1025+69A>T, and c.1025+138C>T) with two of them (c.610A>G and c.916A>G) causing amino acid changes (P.Asn128Ser and P.Val230Ala). There was no statistical association between the variations near the three exons of EDNRB and feline heterochromia iridis (chi-square test, P-value >0.05).

The oral cavity of dogs and cats is colonized by hundreds of bacterial species. Here, we describe the bacterial composition in the dental calculus of dogs and cats. Dental calculus samples from 43 dogs and 4 cats were pooled into four different groups. Dogs were categorized into three groups: non-small breed dogs (NSB), non-brachycephalic small breed dogs (SB) and brachycephalic small breed dogs (SBb). The fourth group included cats. Bacterial communities were identified based on 16S rRNA sequencing (V3 and V4 hypervariable regions) with the Illumina platform. The numbers of operational taxonomic units (OTUs) identified in the three groups of dogs were 180, 190 and 150 and in NSB, SBb and SB, respectively, while in cats there were 111 OTUs. In dental calculus from both dogs and cats, the phylum Firmicutes had the highest proportion of read number, especially the class Clostridia. PCoA and UPGMA analysis revealed differences in the microbiomes of canine and feline calculus. Our findings demonstrated that the bacterial communities in calculus seemed to differ from those in other sites of the oral cavity. Calculus may serve as a potential habitßat for the growth of bacteria linked to canine and feline periodontal disease.

The aims of this study were to investigate the coding sequence and the deduced amino acid sequence of Asian elephant"s biglycan gene as well as its expression in different tissues and conditions using wound healing as a model. The results showed that Asian elephant biglycan coding sequence was 1,110 base pair (bp) long (accession number: JQ753329), encoding 369 amino acids. The coding and amino acid sequences between Asian and African elephants revealed 99% and 98% similarity, respectively. The conserved domains of biglycan protein were also observed. In addition, its expression was found in 15 tissues with a predominant expression in cartilage and spleen. For expression analysis in the wound healing process, it was found that the level of biglycan mRNA was influenced by many factors, including age, type of wound and stage of wound healing.

Abstract The dugong (Dugong dugon, Müller) is an endangered marine mammal species. We examined the relationship between sex, habitat and body length based on the skull and scapular morphology and morphometrics of 81 dugong samples in Thailand. A total of 58 parameters from the skull and scapula (25 from the cranium, 23 from the mandible and 10 from the scapula) as well as tusks were used in this study. Data were analyzed by univariate analysis, followed by discriminant analysis and multivariate linear regression. Here we show, 100% and 98.5% accuracy rates for sexing using large tusks and the skull, respectively. Scapular morphology using the caudal border tubercle and coracoid process showed 91.30% and 96.15% accuracy rates for identifying males and females. Skull morphometrics could categorize dugong habitat, i.e. living in the Andaman Sea or Gulf of Thailand, with 100% accuracy. Moreover, our model could be used to estimate body length with coefficient of determination (R 2) of 0.985. The results of our study showed that skull morphology and morphometric measurements could be used as a tool for sex identification, location identification and estimation of body length. But scapular morphology is the best tool for sex identification in dugongs.

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.

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.

Mineralized tissues accumulate elements that play crucial roles in animal health. Although elemental content of bone, blood and teeth of human and some animal species have been characterized, data for many others are lacking, as well as species comparisons. Here we describe the distribution of elements in horn (Bovidae), antler (Cervidae), teeth and bone (humerus) across a number of species determined by handheld X-ray fluorescence (XRF) to better understand differences and potential biological relevance. A difference in elemental profiles between horns and antlers was observed, possibly due to the outer layer of horns being comprised of keratin, whereas antlers are true bone. Species differences in tissue elemental content may be intrinsic, but also related to feeding habits that contribute to mineral accumulation, particularly for toxic heavy metals. One significant finding was a higher level of iron (Fe) in the humerus bone of elephants compared to other species. This may be an adaptation of the hematopoietic system by distributing Fe throughout the bone rather than the marrow, as elephant humerus lacks a marrow cavity. We also conducted discriminant analysis and found XRF was capable of distinguishing samples from different species, with humerus bone being the best source for species discrimination. For example, we found a 79.2% correct prediction and success rate of 80% for classification between human and non-human humerus bone. These findings show that handheld XRF can serve as an effective tool for the biological study of elemental composition in mineralized tissue samples and may have a forensic application.