The long-tailed goral (Naemorhedus caudatus) is classified as 'vulnerable' by the International Union for Conservation of Nature and has been designated an endangered species requiring conservation and management in Korea. The sex determination region of the Y (SRY) gene is a useful marker for the study of paternal lineages; however, the SRY gene of the goral has not yet been sequenced. In this study, the nucleotide sequence of the SRY gene of long-tailed gorals was determined based on the sequence of the SRY gene of goats (Capra hircus). The obtained sequences were aligned with those of other species in the Bovidae family. The long-tailed goral SRY gene comprised 720 base pairs, and its nucleotide and protein sequences were identical to those of goats, sheep (Ovis aries), and cattle (Bos taurus) by 96%, 97%, and 93%, respectively. The results of phylogenetic insights obtained from this study constitute important references for genetic diversity and pedigrees studies of male long-tailed gorals and closely related species.

The long-tailed goral (Naemorhedus caudatus) is an endangered animal species in all its habitats worldwide, including South Korea. The imbalanced sex ratio in fragmented habitats is closely associated with extinction. Therefore, sex identification using wild animal samples would be necessary. However, only a few studies have been reported about the sex identification of gorals. In this study, we thus aimed at comparing the efficiency of sex identification using various goral sample types as templates and the amelogenin (AMEL) and DEAD-box polypeptide 3 (DDX3) genes as target sequences. We extracted DNA from goral feces, tissues, and blood samples, then amplified the AMEL (SE47/SE48 and SE47/SE53 primer pairs) and DDX3 genes for sex identification, comparing the goral DDX3X and DDX3Y target sequences to those in cattle. Our results indicated that the tissue- and blood sample-derived AMEL amplicons showed an unspecific band pattern containing the sex-specific band in the case of both primer pairs we used, whereas the DDX3 amplicon showed only the sex-specific band. In the case of the feces samples, only the sex-specific band was amplified using both the AMEL and DDX3 primer pairs. However, we found that the DDX3 amplicon exhibited a clearer band pattern than the AMEL amplicon. Then, we compared the DDX3X and DDX3Y target sequences between cattle and gorals. We found 5 and 8 nucleotide differences in the DDX3X and DDX3Y sequences, respectively. In conclusion, the DDX3 gene-related sex identification of the long-tailed goral appears to be more efficient and precise than the AMEL gene-related approach. This method could be used for the sex identification of the members of the Bovidae family.

In Korea, ecological research on the goral is underway to restore and increase population size of these ungulates, but clinical research on gorals species is still in its infancy. Owing to the nature of the gorals’ habitat, several animals live together within limited areas; so, the risk of parasitic infection is very high. In this study, we performed physical examination of 17 gorals undergoing breeding and restoration at the Korean Goral Restoration Center. Weight, ear height, horn length, withers height of the anthropometric data of this study were no differences from other researchers. Blood samples from each species were analyzed. There was no significant difference among individuals in terms each value examined, although some hematological and biochemical values, such as neutrophils, lymphocytes, glucose and amylase showed differences among individuals. The average values of blood tests in gorals ware lower in lymphocytes, blood urea nitrogen, and calcium than in livestock goats, and hemoglobin, mean corpuscular hemoglobin, and glucose were higher in livestock. Helminth eggs were detected in 15 out of 17 gorals by microscopic examination of their fecal samples, and Eimeria spp. and Strongyloides spp. were detected in all the gorals. However, there may be cases of death due to diarrhea, which may lead to a decrease in growth and production. Therefore, proper prevention and treatment are needed. These results may serve as an important reference for clinical diagnosis and treatment of gorals. This information helps further understand the current health status of these species and may contribute to a more systematic restoration of the gorals.

The aim of this study was to estimate of ovulation time and parturition day at the same time as breeding in small dog by vaginal cytology and to confirm the accuracy by comparing the expected parturition day and the real one. Characteristic features of vaginal cytology during the estrous cycle were the high proportion of large intermediate cell, superficial cell, anuclear cell and erythrocyte in proestrus, superficial cell and anuclear cell in estrus, parabasal cell, small intermediate cell, large intermediate cell and leukocyte in diestrus, parabasal cell and small intermediate in anestrus, respectively. When day 0 was the parturition day, the period of pregnancy is 67.45(64～75) days when the cornification index (CI) is over 90%. Also, on the basis of ovulation day, 63.65(59～66) days was confirmed, and 57.0(52～60) days was confirmed based on the first day of diestrus. There are the gap of 4 days between the day being over 90% in CI and ovulation day. On the basis of this, when expecting parturition day based on the day being over 90% in CI by vaginal cytology, 18.1% was produced in the same of the expected parturition day and the real one, 30.3% and 33.3% were produced in the gap of one day and two days, respectively so, the accuracy within two days was 81.7%. In addition, based on the first day of diestrus, it also was identified to 81.7% as the difference between the expected parturition day and the real one within 2 days. It demonstrated there are any difference between any expected parturition day by vaginal cytology. Thus on the basis of the day of being over 90% CI, it is fully thought to using clinically due to the possibility of prediction the parturition day at the same time as the determination of the proper time of the optimal mating time.

The purpose of this study is to suggest developing plans for 6 beef brands in Kangwon-do. Data were collected by consumer survey on 12th August, 2010 at Daemyung Resort in Hongcheongun. A total of 658 questionnaires were distributed and collected. SAS 9.1 and Excel 2007 were used as statistical packages and ordered logit model was used as a model to analyze the data. From the results of the study the following improvement plans were suggested for the 6 beef brands in Kangwon-do. First, the 6 beef brand agents should use public relation to give information for consumers. Clean image of Kangewon-do and clear distribution process should be stressed for the public relation. Second, promotion is an important tool to discriminate the 6 beef brands with other beef brands. The discrimination by the 6 beef brands should be focused on taste of the 6 beef brands. Third, market segmentation strategies are needed. The market segmentation may be achieved by segmentation of sales places.

Somatic cells such as oviduct epithelial cell, uterine epithelial cell, cumulus-granulosa cell and buffalo rat river cell has been used to establish an effective culture system for bovine embryos produced in in vitro. But nitric oxide (NO) metabolites secreted from somatic cells were largely arrested the development of bovine in vitro matured/ in vitro fertilized (IVM/IVF) embryos, suggesting that NO was induced the embryonic toxic substance into culture medium. The objective of this study was to investigate whether BOEC co-culture system can ameliorate the NO-mediated oxidative stress in the culture of bovine IVM/IVF embryos. Therefore, we evaluated the developmental rate of bovine IVM/IVF embryos under BOEC co-culture system in the presence or absence of sodium nitroprusside (SNP), as a NO donor, and also detected the expression of growth factor (TGF-p , EGF and IGFBP) and apoptosis (Caspase-3, Bax and Bcl-2) genes. The supplement of SNP over 5 uM was strongly inhibited blastocyst development of bovine IVM/IVF embryos than in control and 1 uM SNP group (Table 2). The developmental rates beyond morulae stages of bovine IVM/IVF embryos co-cultured with BOEC regardless of SNP supplement (40.4% in 5 uM SNP+ BOEC group and 65.1% in BOEC group) were significantly increased than those of control (35.0%) and SNP single treatment group (23.3%, p<0.05: Table 3). The transcripts of Bax and Caspase-3 genes were detected in all experiment groups (1:Isolated fresh cell (IFC), 2:Primary culture cell (PCC), 3:PCC after using the embryo culture, 4: PCC containing 5 uM SNP and 5: PCC containing 5 uM SNP after using the embryo culture), but Bcl-2 gene was not detected in IFC and PCC (Fig. 1). In the expression of growth factor genes, TGF-p gene was found in all experimental groups, and EGF and IGFBP genes were not found in IFC and PCC (Fig. 2). These results indicate that BOEC co-culture system can increase the development beyond morula stages of bovine IVM/IVF embryos, possibly suggesting the alleviation of embryonic toxic substance like nitric oxide.

The objective of this study was to establish the optimal conditions for hypoosmotic swelling (HOS) test to assess the functional integrity of the membranes of boar fresh or frozen/thawed spermatozoa. When pooled semen sample was incubated for 30 min at 37℃ with different test solution of varied osmolarity, the highest percentage of HOS positive spermatozoa was observed in a 150 mOsmol fructose/Na-citrate solution (33.6%). Incubation time did not affect significantly the score of HOS positive spermatozoa observed in a 150 mOsmol fructose/Na-citrate solution at 37℃, but the osmolarity affected the score of HOS positive spermatozoa under the same condition above. Fresh semen was significantly better than frozen/thawed semen in semen parameters evaluated such as motility, viability, membrane integrity and lipid peroxidation (p<0.05). In the relationships of sperm parameters, motility vs viability, motility vs membrane integrity and viability vs membrane integrity were positively correlated (0.82～0.94) but lipid peroxidation vs other estimated factors was negatively correlated (－0.90～－0.98). Among the evaluation methods, motility vs viability, motility vs membrane integrity and lipid peroxidation vs other estimated factors were significantly correlated (p<0.05). These results of this study indicate that the optimal condition of HOST in boar spermatozoa is a 150 mOsmol fructose/Na-citrate solution for 30 min incubation at 37℃ and HOST can substitute the examination of motility, viability and lipid peroxidation.

Astaxanthin is a kind of carotenoid compounds, having a antioxidant and anti-inflammatory activities. The antioxidative mechanism by which carotenoid scavenge free radicals has been clearly elucidated, but has not tried for the development of mammalian preimplantation embryo. This study was conducted to investigate the antioxidative effect of astaxanthin on in vitro development of porcine in vitro fertilized embryos. Porcine embryos derived from in vitro fertilization (IVF) were cultured in 5% CO2 in air at 38.5℃ in PZM-3 medium supplemented with different dosages of astaxanthin (0, 1, 5 and 10 M) and taurine (0, 1, 2.5 and 5 mM) as a positive control, and execute to compare the effects of various antioxidants such as taurine, melatonin and asculatin on in vitro development. The proportions of embryos developed to the blastocyst stage were increased when 1 and 5 M of astaxanthin (26.6 and 23.4%, respectively) and 1 and 2.5 mM taurine (25.8 and 26.4%, respectively) were supplemented, compared to controls (p<0.05). Also, various antioxidant-treated groups were significantly higher rates of blastocysts (astaxanthin, 27.4%; taurine, 29.1%; melatonin, 26.8%; aesculetin, 27.9%, respectively ) than control (18.8%). There was no difference in mean cell number of blastocysts between antioxidants and control. This result indicates that astaxanthin has an antioxidant feature when porcine IVF embryos were cultured in vitro.

본 연구는 내분비계 장애물질인 BPA와 DEHP가 돼지 정액 성상(운동성, 생존율, 원형질막의 정상성, 기형율)에 유해한 영향을 미치는지를 검토하였다. 일정 농도로 희석한 돼지 정액에 BPA와 DEHP를 각각 0, 1, 10, 100 μM의 농도로 처리하여 3, 6, 9시간 동안 체외배양을 실시하였다. BPA 처리에 따른 운동성과 생존율은 체외배양 시간과 첨가농도에 따라 감소하였으며, 체외배양시간이 경과함에 따라 대조구와 명백한 차이를 나타냈다. 100 μM의 농도로 처리한 경우의 운동성과 생존율은 체외배양시간에 관계없이 처리군이 대조구에 비해 유의적으로 감소하였다(p<0.05). DEHP 처리에 따른 정자의 운동성과 생존율도 체외배양시간과 첨가농도에 따라 감소하여 BPA의 성적과 유사한 경향을 나타냈다. 정자원형질막의 기능성은 배양시간에 따라 감소하였으며 BPA 및 DEHP 첨가농도 간에 차이가 인정되었으며, 특히 100 μM 처리구에서 급격히 감소하는 성적을 나타났다. 정자의 기형율은 BPA 및 DEHP 처리시간 및 농도에 크게 영향을 받지는 않았다. 본 연구의 결과로 볼 때, 고농도의 BPA와 DEHP(>10 μM)에 정자의 장시간 노출은 유해한 영향을 줄 것으로 사료된다.