Life History of a Unique Asian Plethodontid Salamander, Karsenia koreana.

Using preserved specimens, we studied the basic life history of the topotypic population of the unique Asian plethodontid salamander, Karsenia koreana. Of 51 individuals examined, 11 males and 13 females were judged as mature from the development of gonads. The ovarian eggs were large (diameter 3.7-4.8 mm) and yellow to orange in color, and the clutch size was about 8-10. These values approximate those of actually spawned eggs recently reported. Skeletochronological analyses revealed the average age of males (5.3 years) to be lower than females (7.3 years). The age at maturity and maximum observed longevity were four and nine years in males and five and 10 years in females, respectively. In the growth curves estimated by a von Bertalanffy growth model, the growth coefficient and asymptotic SVL did not differ between the sexes, although males (40.6 mm) were smaller than females (45.3 mm) in the average snout-vent length. The time and place of courtship behavior, oval development, hatching, and especially, whether the species shows aquatic larval stage or direct development, are important topics to be resolved in future.

Detection and Molecular Characterization of Cryptosporidium spp. from Wild Rodents and Insectivores in South Korea.

In order to examine the prevalence of Cryptosporidium infection in wild rodents and insectivores of South Korea and to assess their potential role as a source of human cryptosporidiosis, a total of 199 wild rodents and insectivore specimens were collected from 10 regions of South Korea and screened for Cryptosporidium infection over a period of 2 years (2012-2013). A nested-PCR amplification of Cryptosporidium oocyst wall protein (COWP) gene fragment revealed an overall prevalence of 34.2% (68/199). The sequence analysis of 18S rRNA gene locus of Cryptosporidium was performed from the fecal and cecum samples that tested positive by COWP amplification PCR. As a result, we identified 4 species/genotypes; chipmunk genotype I, cervine genotype I, C. muris, and a new genotype which is closely related to the bear genotype. The new genotype isolated from 12 Apodemus agrarius and 2 Apodemus chejuensis was not previously identified as known species or genotype, and therefore, it is supposed to be a novel genotype. In addition, the host spectrum of Cryptosporidium was extended to A. agrarius and Crosidura lasiura, which had not been reported before. In this study, we found that the Korean wild rodents and insectivores were infected with various Cryptosporidium spp. with large intra-genotypic variationa, indicating that they may function as potential reservoirs transmitting zoonotic Cryptosporidium to livestock and humans.

Spatiotemporal Niche Separation among Passeriformes in the Halla Mountain Wetland of Jeju, Republic of Korea: Insights from Camera Trap Data.

This study analyzed 5322 camera trap photographs from Halla Mountain Wetland, documenting 1427 independent bird sightings of 26 families and 49 species of Passeriformes. Key observations include morning activities in Cyanoptila cyanomelana and Horornis canturians and afternoon activity in Muscicapa dauurica and Phoenicurus auroreus. Wetlands were significantly preferred (P_i = 0.398) despite their smaller area, contrasting with underutilized grasslands (P_i = 0.181). Seasonal activity variations were notable, with overlap coefficients ranging from 0.08 to 0.81 across species, indicating diverse strategies in resource utilization and thermoregulation. Population density was found to be a critical factor in habitat usage, with high-density species showing more consistent activity patterns. The study's results demonstrate the ecological adaptability of Passeriformes in the Halla Mountain Wetland while highlighting the limitations of camera trapping methods. These limitations include their fixed field of view and intermittent recording capability, which may not fully capture the spectrum of complex avian behaviors. This research underlines the need for future studies integrating various methodologies, such as direct observation and acoustic monitoring, to gain a more comprehensive understanding of avian ecology.

Decreased tissue and serum expression of galectin-7 in patients with hypertrophic scars.

Hypertrophic scars (HS) result from an imbalance between collagen biosynthesis and matrix degradation during wound healing. In this study a proteomics approach was used to compare the protein profiles of skin tissue obtained from patients with HS and healthy controls. One of the epidermal proteins, galectin-7 was markedly down-regulated in HS. Serum levels of galectin-7 in 27 patients with HS were less than 1/3 of those in 15 healthy controls. Tissue protein expression was subsequently evaluated using immunohistochemical staining on HS tissue and on serially-obtained control tissue during wound healing. Weaker galectin-7 immunoreactivity was detected along the cytoplasmic membrane of basal and suprabasal cells in samples from HS. In addition, galectin-7 was stained in the extracellular space of the upper papillary dermis in HS tissue. Ablative laser treatment, used to induce wound healing of healthy control tissue, demonstrated marked galectin-7 expression at the cytoplasmic membrane on days 3, 5, 14 and 21. Pronounced galectin-7 staining at the upper papillary dermis was detected on days 1, 3 and 10. These results suggest that the differences in galectin-7 expression and subcellular and extracellular distribution may be crucially involved in the pathogenic process of HS.

Porcine SPP1 gene polymorphism association with phenotypic traits in the Landrace × Jeju (Korea) Black pig F2 population.

This study examined polymorphisms of the secreted phosphoprotein 1 (SPP1) gene and its association with growth and carcass traits in the F(2) population of the crossbred Landrace × Jeju (Korea) Black pig. The authors detected the presence/absence polymorphisms of short interspersed nuclear element in the SPP1 intron 6 of the population; they then designated the longer fragment as allele A and the shorter one as allele B. The SPP1 A/B heterozygous pigs evidenced significantly heavier body weight at birth and on days 21 and 70, and a higher level of average daily gain during the early developmental period than was seen in the A/A and B/B homozygous pigs (P < 0.05). Further, the SPP1 A/B heterozygous pigs evidenced significantly greater body length, less backfat thickness measured at three different sites, and larger loin muscle area than the homozygotes (P < 0.05). On the other hand, the levels of late average daily gain, 140th-day body weight, and marbling score were not significantly associated (P > 0.05). The results of this study reveal faster growth rate and differences in pig productivity according to genotypes of the SPP1 gene. These findings demonstrate that SPP1 genotypes may effectively function as molecular genetic markers for the improvement of Jeju Black pig-related crossbreeding systems.

A promoter polymorphism of MSTN g.-371T>A and its associations with carcass traits in Korean cattle.

A promoter polymorphism of bovine Myostatin (MSTN) gene g.-371T>A was screened in Holstein and two Korean indigenous cattle breeds, Hanwoo and Jeju Black cattle (JBC). The MSTN g.-371T>A polymorphism was found in all three cattle breeds tested. An allele MSTN g.-371A was the most frequent in the JBC breed among breeds tested. The association of MSTN genotypes for carcass traits was also tested in the Hanwoo population. Significant differences were found between the genotypes and level of meat quality grade index which converted the marbling score levels (P < 0.05), reflecting the metabolic role of MSTN for inhibition of preadipocyte differentiation in intramuscular fat deposition. In addition, significant differences were found for fat color index of backfat according to MSTN genotypes (P < 0.05), suggesting that MSTN may play a role in deposition of white-yellow adipocytes in backfat. However, there was no detection of significant association of genotypes with the live weight, carcass weight, backfat thickness, eye muscle area, marbling score, or meat color index (P > 0.05). Despite the lack of statistical association, wild type g.-371T/-showed association patterns similar to those of A/A homozygotes, such as heavier weights, thinner backfat, larger eye muscle area, and lower marbling score. The results of the present study suggest that MSTN promoter polymorphism g.-371T>A may affect carcass traits, which could be a useful molecular marker for planning improvements in the economic traits of Korean cattle breeds.

Molecular Evidence Reveals the Sympatric Distribution of Cervus nippon yakushimae and Cervus nippon taiouanus on Jeju Island, South Korea.

Non-native species threaten native ecosystems and species, particularly on islands where rates of endemism and vulnerability to threats are high. Understanding species invasion will aid in providing insights into ecological and evolutionary processes. To identify the non-native sika deer (Cervus nippon) population in Jeju, South Korea, and their phylogenetic affinities, we collected tissue samples from roadkill and the World Natural Heritage Headquarters in Jeju. Mitochondrial DNA cytochrome B (CytB) gene sequences were analyzed to determine two distinct CytB haplotypes. Phylogenetic analysis using maximum likelihood tree revealed two haplotypes of CytB clustered into two different groups representing two subspecies: C. n. yakushimae, native to Japan, and C. n. taiouanus, native to Taiwan. The tentative divergence time between the two subspecies was estimated at 1.81 million years. Our study confirmed that the two subspecies of sika deer are sympatric in the natural ecosystem of Jeju Island. This study provides valuable information to help government and conservation agencies understand alien species and determine control policies for conserving native biodiversity in South Korea.

Systematic relationships of Oriental tiny frogs of the family Microhylidae (Amphibia, Anura) as revealed by mtDNA genealogy.

We estimated the genealogical relationships and assessed systematic relationships among 45 out of 89 named species and four unnamed taxa from 11 of 14 genera of the Oriental microhylids from 1767 bp sequences of the mitochondrial DNA genes 12S rRNA and 16S rRNA using maximum parsimony, maximum likelihood, and Bayesian inference methods. Monophyly was rejected for the subfamily Microhylinae, and our data reveal four well-supported clades whose relationships to each other are unresolved: (A) Microhyla, Calluella, and Glyphoglossus, (B) Chaperina, (C) Kaloula, Phrynella, and Metaphrynella, and (D) Micryletta. They were genetically as divergent from each other as from another Oriental subfamily Kalophryninae, and could be recognized as distinct subfamilies. Within Clade A, our data reveal three well-supported subclades whose relationships to each other are unresolved: (AI) Microhyla-I, (AII) Calluella and Glyphoglossus, and (AIII) Microhyla-II. Of the two enigmatic Malaysian genera, whose subfamilial placement has been undetermined, Phrynella was found to be the sister species of Metaphrynella in Clade C, whereas Gastrophrynoides was grouped in the Papua-Australian subfamily Asterophryinae. Currently recognized subgenera and species groups within Microhyla based on morphology were not supported phylogenetically, and require thorough reassessments.

The Jeju weasel, Mustela sibilica quelpartis, a new definitive host for Gnathostoma nipponicum Yamaguti, 1941.

Adult gnathostomes were discovered in the stomach of the Jeju weasel, Mustela sibilica quelpartis, road-killed in Jeju-do (Province). Their morphological characters were examined to identify the species. Total 50 gnathostome adults were collected from 6 out of 10 weasels examined. In infected weasels, 4-6 worms were grouped and embedded in each granulomatous gastric tumor, except 1 weasel. Male worms were 25.0×1.4 mm in average size, and had a tail with pedunculate papillae, a spicule, and minute tegumental spines. Females were 40.0×2.5 mm in average size, and had a tail without tegumental spines. Pointed and posteriorly curved hooklets were arranged in 8-10 rows on the head bulb. Tegumental spines were distributed from behind the head bulb to the middle portion of the body. The spines were different in size and shape by the distribution level of the body surface. Fertilized eggs were 65.5×38.9 µm in average size, and had a mucoid plug at 1 pole. These gnathostomes from Jeju weasels were identified as Gnathostoma nipponicum Yamaguti, 1941. By the present study, it was confirmed for the first time that G. nipponicum is distributed in Jeju-do, the Republic of Korea, and the Jeju weasel, M. sibilica quelpartis, plays a crucial role for its definitive host.

Discovery of larval Gnathostoma nipponicum in frogs and snakes from Jeju-do (Province), Republic of Korea.

A survey was performed to find out the intermediate hosts of Gnathostoma nipponicum in Jeju-do (Province), the Republic of Korea. In August 2009 and 2010, a total of 82 tadpoles, 23 black-spotted pond frogs (Rana nigromaculata), 7 tiger keelback snakes (Rhabdophis tigrinus tigrinus), 6 red-tongue viper snakes (Agkistrodon ussuriensis), and 2 cat snakes (Elaphe dione) were collected in Jeju-do and examined by the pepsin-HCl digestion method. Total 5 gnathostome larvae were detected in 3 (50%) of 6 A. ussuriensis, 70 larvae in 3 of 7 (42.9%) R. tigrinus tigrinus, and 2 larvae in 2 of 82 (8.7%) frogs. No gnathostome larvae were detected in tadpoles and cat snakes. The larvae detected were a single species, and 2.17 × 0.22 mm in average size. They had characteristic head bulbs, muscular esophagus, and 4 cervical sacs. Three rows of hooklets were arranged in the head bulbs, and the number of hooklets in each row was 29, 33, and 36 posteriorly. All these characters were consistent with the advanced third-stage larvae of G. nipponicum. It has been first confirmed in Jeju-do that R. nigromaculata, A. ussuriensis, and R. tigrinus tigrinus play a role for intermediate and/or paratenic hosts for G. nipponicum.

Dispersal history of Miniopterus fuliginosus bats and their associated viruses in east Asia.

In this study, we examined the role of the eastern bent-winged bat (Miniopterus fuliginosus) in the dispersion of bat adenovirus and bat alphacoronavirus in east Asia, considering their gene flows and divergence times (based on deep-sequencing data), using bat fecal guano samples. Bats in China moved to Jeju Island and/or Taiwan in the last 20,000 years via the Korean Peninsula and/or Japan. The phylogenies of host mitochondrial D-loop DNA was not significantly congruent with those of bat adenovirus (m2XY = 0.07, p = 0.08), and bat alphacoronavirus (m2XY = 0.48, p = 0.20). We estimate that the first divergence time of bats carrying bat adenovirus in five caves studied (designated as K1, K2, JJ, N2, and F3) occurred approximately 3.17 million years ago. In contrast, the first divergence time of bat adenovirus among bats in the 5 caves was estimated to be approximately 224.32 years ago. The first divergence time of bats in caves CH, JJ, WY, N2, F1, F2, and F3 harboring bat alphacoronavirus was estimated to be 1.59 million years ago. The first divergence time of bat alphacoronavirus among the 7 caves was estimated to be approximately 2,596.92 years ago. The origin of bat adenovirus remains unclear, whereas our findings suggest that bat alphacoronavirus originated in Japan. Surprisingly, bat adenovirus and bat alphacoronavirus appeared to diverge substantially over the last 100 years, even though our gene-flow data indicate that the eastern bent-winged bat serves as an important natural reservoir of both viruses.

SINE indel polymorphism of AGL gene and association with growth and carcass traits in Landrace x Jeju Black pig F(2) population.

Genetic polymorphisms in the glycogen debrancher enzyme (AGL) gene were assessed with regard to their association with growth and carcass traits in the F(2) population crossbred Landrace and Jeju (Korea) Black pig. Three genotypes representing the insertion and/or deletion (indel) polymorphisms of short interspersed nuclear element were detected at frequencies of 0.278 (L/L), 0.479 (L/S), and 0.243 (S/S), respectively. The AGL S allele-containing pigs evidenced significantly heavier body weights at birth, the 3rd week, 10th week, and 20th week during developmental stages and higher average daily gains during the late period than were noted in the L/L homozygous pigs (P < 0.05), respectively. However, average daily gains during the early period were not significantly associated with genotype distribution (P > 0.05). With regard to the carcass traits, the S allele pigs (S/-) evidenced significantly heavier carcass weights and thicker backfat than was measured in L/L homozygous pigs (P < 0.05). However, body lengths, meat color, and marbling scores were all found not to be statistically significant (P > 0.05). Consequently, the faster growth rate during the late period and backfat deposition rather than intramuscular fat deposition cause differences in pig productivity according to genotypes of the AGL gene. These findings indicate that the AGL genotypes may prove to be useful genetic markers for the improvement of Jeju Black pig-related crossbreeding systems.

Length difference between equine ZFX and ZFY genes and its application for molecular sex determination.

PURPOSE: we analyzed the sex chromosome-encoding ZFX-ZFY genes and tested molecular sexing using the amplification patterns of intron 9 of ZFX-ZFY in the horse. METHODS AND RESULTS: the amplification of the ZFX-ZFY produced two distinct patterns, reflecting sexual dimorphism based on a length difference between the X and Y chromosomes. The amplification products from foals showed two distinct bands: one was common to all foals and mares, indicating that this band was amplified from ZFX, while the other was specific to some foals, indicating that it was from ZFY. The result based on the PCR assay was identical to the results of amplification of the Y chromosome-specific SRY gene and those of investigations of the phenotypic gender in three different horse populations. CONCLUSION: we suggest that this PCR strategy for determining sexes by comparing the amplification patterns of ZFX-ZFY genes is a convenient and precise method for discriminating sexes in horses.

Illegal wildlife trade in local markets of Feuang and Mad districts of Vientiane Province, Lao People's Democratic Republic.

The Lao People's Democratic Republic has emerged as a hub for illegal flora and fauna trade, more specifically, species in protected categories. However, numerous local species are traded and given less consideration despite their importance. Hence, we observed the local markets in the Feuang and Mad districts of Vientiane Province in summer and winter seasons to determine the species for trade, as well as their volume and conservation status. Altogether, 602 specimens corresponding to 23 genera and 22 species were identified. Among them, the highest number of species was mammals, followed by birds, reptiles, amphibians, and insects. Six species are listed in threatened categories according to the International Union for Conservation of Nature and nine under Convention on International Trade in Endangered Species categories. The species in trade varied between seasons, as winter was the main poaching season for mammals and birds and summer was for amphibians, reptiles, and insects. This study revealed that food, traditional medicine, and curio production were the main reasons for wildlife trade. The lack of strong regulations, monitoring and law enforcement, and poverty are the major reasons for wildlife trade. Therefore, strong law enforcement, creating alternate income sources, and participatory conservation programs are required to effectively control wildlife trade in the Lao People's Democratic Republic.

Various branching patterns of the aortic arch in the Siberian roe deer (Capreolus pygargus, Pallas, 1771) on Jeju Island.

This study examined and compared the branching pattern of the aortic arch (AA) and its major branches in the Siberian roe deer (Capreolus pygargus) on Jeju Island (Jeju roe deer [JRD]) with those in the roe deer of the Korean peninsula (mainland roe deer [MRD]). Seven of the nine expected types was observed in the arterial silicone casts of 29 deer (10 males, 19 females). The JRD was identical to the MRD in that absence of the typical pattern; however, the main three pattern types differed between the two. This difference resulted from differences in the branching patterns of the right subclavian artery and costocervical trunk. In conclusion, the JRD has different type of AA from the MRD.

Encephalomyocarditis virus is potentially derived from eastern bent-wing bats living in East Asian countries.

Bats are reservoir hosts of many zoonotic viruses and identification of viruses that they carry is important. This study aimed to use high throughput screening to identify the viruses in fecal guano of Taiwanese insectivorous bats caves in order to obtain more information on bat-derived pathogenic viruses in East Asia. Guano samples were collected from two caves in Taiwan, pooled, and then subjected to Multiplex PCR-based next generation sequencing for viral identification. Subsequently, encephalomyocarditis virus (EMCV) sequence was detected and confirmed by reverse transcription PCR. EMCV is considered as rodent virus and thus, animal species identification through cytochrome oxidase I (COI) barcoding was further done to identify the viral source. Finally, determination of distribution and verification of the presence of EMCV in guano obtained from Japanese and South Korean caves was also done. We concluded that the guano collected was not contaminated with the excrement of rodents which were reported and presumed to live in Taiwan. Also, EMCV genome fragments were found in guanos of Japanese and South Korean caves. It is possible that the eastern bent-wing bat (Miniopterus fuliginosus) is one of the natural hosts of EMCV in East Asia.

Genetic identification for prey birds of the Endangered peregrine falcon (Falco peregrinus).

To reveal the dietary ecology of the peregrine falcon (Falco peregrinus), we conducted a molecular analysis for mitochondrial genes (COI and CYTB) to identify the prey species collected from a feeding place found on Sasu Island, South Korea. The results from noninvasive genetic analysis showed that the sequences obtained came from nine species of birds (Cuculus canorus, Eurystomus orientalis, Limosa limosa, Microscelis amaurotis, Oriolus chinensis, Phasianus colchicus, Sterna hirundo, Streptopelia orientalis, and Turdus pallidus). Five of the species (C. canorus, M. amaurotis, S. hirundo, S. orientalis, and T. pallidus) had previously been observed on this island, but the other four species (E. orientalis, L. limosa, O. chinensis, and P. colchicus) were newly identified as present. No mtDNA sequences of land animals such as amphibians, reptiles, and mammals were found in the dietary remains, suggesting that the peregrine falcon preys mostly on other birds rather than on other animals inhabiting Sasu Island. This island has rich avian diversity and abundant animal populations and therefore supplies sufficient dietary resources for the peregrine falcon. Our findings suggested that a DNA-based molecular method may be useful to identify the prey species of these birds and may be valuable in future studies of the Endangered peregrine falcon.

Genetic identification of prey species from teeth in faeces from the Endangered leopard cat Prionailurus bengalensis using mitochondrial cytochrome b gene sequence.

To understand the dietary ecology of the leopard cat (Prionailurus bengalensis), DNA analysis was performed to identify prey species using DNA isolated from teeth harvested from the faeces of this feline species. From 70 DNA samples, a total of 52 mitochondrial DNA (mtDNA) cytochrome b (cytb) gene sequences of mammals were identified. The results of a sequence identity test indicated that those sequences were derived from four rodent species (Apodemus agrarius, Apodemus peninsulae, Eothenomys regulus and Tamias sibiricus) and two shrew species (Crocidura lasiura and Crocidura shantungensis). The sequences contained nine unique cytb sequences from site 1 and 13 from site 2. These results indicate that the leopard cat hunts rodents and shrews, and at least nine animals at site 1 and 13 animals at site 2 were eaten. These findings suggest that the animal molecular signatures that remain undigested in the faeces may provide useful ecological information about food items and may contribute to a better understanding of the leopard cat's feeding ecology.

New record of the Oriental house rat, Rattus tanezumi, in Nepal inferred from mitochondrial Cytochrome B gene sequences.

This study determines the presence of R. tanezumi from in Nepal using morphological and molecular analyses. Morphologically, it is indistinguishable with R. rattus owing to similar fur colour and morphometric data. However, molecular identification and phylogenetic analysis using sequences of the mitochondrial DNA (mtDNA) Cytochrome B (CytB) gene revealed two different species R. rattus and R. tanezumi from collected specimens. The genetic distance between R. rattus and R. tanezumi was found 0.043. In phylogenetic tree, the clade of R. tanezumi is distinguished into two sub-clades, R. tanezumi found in Nepal, and East Asian countries, China, Laos, Thailand, Viet Nam, and South Korea have genetic distance 0.031, suggesting the different lineages of R. tanezumi. This study confirmed the R. tanezumi present in Nepal. Our findings suggest that morphological analysis and molecular study should be carried out simultaneously for accurate identification of small sized cryptic mammals like R. tanezumi and R. rattus.

Branching patterns of the aortic arch in the Siberian roe deer (Capreolus pygargus Pallas, 1771).

This study examined the branching pattern of the aortic arch (AA) and its major branches in the Siberian roe deer (Capreolus pygargus Pallas, 1771) from South Korea. A total of eight of the nine expected types, based on the branching site and bilateral levels of the costocervical trunk (CCT) and subclavian artery (SB), were observed in the arterial silicone casts of 35 deer (16 males, 19 females). This deer has no typical type. The three most common types were present in 28.6, 25.7 and 20.0% of cases and resulted from different branching patterns of the left CCT and left SB. These results suggest that the Siberian roe deer in the Korean peninsula has various AA branching patterns, which differs from other ruminants.