A study was performed to investigate the effect of reseeding rates of Italian ryegrass (IRG)〔Lolium multiflorum L.〕on its nutritive value and productivity in native grasssland of Jeju region from Oct. 2011 to Jul. 2012. IRG was sown at reseeding rates (0 30, 40 and 50 kg/ha) in native grassland. When IRG was sown, there was no draught and lodging damage. Plant height in treatment 30 kg/ha of reseeding IRG was the tallest at all harvest stages (p<0.05). DM yield in treatment of 30 kg/ha of reseeding IRG was 12,790 kg/ha and increased 2.7 folds as compared to control. However, there was no significant difference among the treatments. The content of neutral detergent fiber (NDF) significantly decreased in treatment of reseeding, compared to control (p<0.05). In general, the content of NDF in second harvest stage was lower than that of control in first harvest stage. The content of acid detergent fiber (ADF) also showed simliar trend as content of NDF. The content of total digestible nutrients (TDN) in treatments of 30 kg/ha and 50 kg/ha of reseeding were approximately 45.2% each. The content of TDN in treatments of 30 kg/ha and 50kg/ha of reseeding increased as compared to 40 kg/ha and control of reseeding (p<0.05). Therefore, it is conclued that reseeding rate of IRG may improve the yield and nutritive values of IRG at 30 kg/ha.

Precise, rapid and simple methods for species identification in animals are among the most important techniques in the livestock industry and research fields including meat classification. In this study, polymerase chain reaction (PCR) based molecular identification using inter species polymorphisms were examined by PCR-restriction fragment length polymorphism (RFLP) analysis for mitochondrial DNA (mtDNA) cytochrome b (CYTB) gene sequences among four mammalian livestock animals (cattle, horse, goat and pig). The results from PCR-RFLP analysis using the AluI restriction enzyme were also provided for the species-specific band patterns among CYTB gene sequences in these four species. The AluI-digestion for CYTB genes provided interesting migration patterns differentially displayed according to each species. Cattle and horse had one AluI-recognition site at different nucleotide positions and their AluI-digested fragments showed different band patterns on the gels. Pig had two AluI-recognition sites within the amplified CYTB sequences and produced three bands on the gels. Goat had no AluI-recognition site and was located at the same position as the uncut PCR product. The results showed the species-specific band patterns on a single gel among the four livestock animal species by AluI-RFLP. In addition, the results from blind tests for the meat samples collected from providers without any records showed the identical information on the species recorded by observing their phenotypes before slaughter. The application of this PCR-RFLP method can be useful and provide rapid, simple, and clear information regarding species identification for various tissue samples originating from tested livestock species.

The objective of the study is to assess temperament of a horse based on general temperament test by a questionnaire survey. Five test criteria were identified: gentleness, patience, aggressiveness, sensitivity, and friendliness, each on a 5-point scale. 114 horses bred at the National Institute of Animal Science (NIAS) in 2010, 2011 and 2012. The horses recorded scores of 3.6∼3.9 for gentleness, 3.1∼3.6 for patience, 3.4∼4.0 for aggressiveness, 2.8∼3.2 for sensitivity, and 3.4∼3.8 for friendliness, the overall score for sensitivity the lowest. Horses born in 2012 scored lower than the rest in all five areas at a statistically significant level (P<0.05). By gender, the colts scored higher than the fillies in all five areas, but the discrepancy was not statistically significant. Factor analysis yielded only one factor, and the Cronbach's α value was 0.980 for standardization of Factor 1, indicating a high reliability of internal consistency. The correlation coefficients among the test criteria ranged between 0.85 and 0.91 (P<0.01). The assessment criteria used in this study are expected to provide a useful basis designing a temperament test horses.

This study was carried out to examine a molecular marker system for parentage test in Jeju Black cattle (JBC). Based on the preliminarily studies, we finally selected for construction of a novel genetic marker system for molecular traceability, identity test, breed certification, and parentage test in JBC and its related industrial populations. The genetic marker system had eight MS markers, five indel markers, and two single nucleotide polymorphisms (SNPs; g.G299T and g.del310G) within MC1R gene which is critical to verify the breed specific genotypes for coat color of JBC differing from those of exotic black cattle breeds such as Holstein and Angus. The results showed lower level of a combined non-exclusion probability for second parent (NE-P2) of 4.1202×10-4 than those previously recommended by International Society of Animal Genetics (ISAG) of 5.000×10-4 for parentage, and a combined non-exclusion probability for sib identity (NE-SI) of 2.679×10-5. Parentage analysis has been successfully identified the JBC offspring in the indigenous population and cattle farms used the certified AI semens for production using the JBC-derived offspring for commercial beef. This combined molecular marker system will be helpful to supply genetic information for parentage test and traceability and to develop the molecular breeding system for improvement of animal productivity in JBC population.

This study was carried out to investigate synthetic extender for semen cryopreservation of Jeju Native Black Bull. The semen was collected using an artificial vagina and transported to the laboratory. The semen was diluted 1:1 by Tris-Egg yolk extender and contrifuged in 1,500 rpm for 15 minutes. The supernatant was removed. The pellect was diluted to final sperm concentration of 2×108/ml by doubling in every 30 minutes at 4℃ cold chamber. The semen was equilibrated for 4 hours at cold chamber and packed to 0.5 ml straw. The semen straws were located above 5 cm for 10 minutes. The height and duration affect the freezing speed by temperature. The frozen straw was plunged to LN2. The presented straws were examined the viability and motility after thawed at 37℃ water bath. Frozen-thawed sperm were evaluated sperm viability, membrane integrity and acrosome integrity. Post-thawed sperm viability has been significantly higher (p<0.05) in fresh sperm (93.27±1.62%) than frozen-thawed sperm (73.34±3.27%). However, there were no significant differences between fresh and frozen-thawed dead cell rate (7.35±2.63 vs, 13.71±2.85). In sperm motility, between Triladyl and AndroMed Extender, there was no significant different (72.86±2.83 vs, 81.47±2.48), similarly, the dead cell rates was similar (18.41±3.42% and 17.26±4.25). The results of our study suggest that AndroMed to the freezing extender showed more positive effect on the frozen-thawed spermatozoa in Jeju Native Black bull semen.

The present study was to assess the in vitro viability and sexing rate of bovine embryos. Blastocysts were harvested on day 7～9 day after insemination(in vitro and in vivo), and the sex of the embryos was examined using the LAMP method. Embryo cell biopsy was carried out in a 80 μl drop Ca2+, Mg2+ free D-PBS and, biopsied embryos viability were evaluated after more 12 h culture in IVMD culture medium. The formation of recovered embryo to expanded and hatching stages had ensued in higher of sexed embryo in vivo than in vitro (100% vs. 89%, p<0.05), and in vitro, the rates of degeneration after sexing were significantly (p<0.05) higher in vitro than in vivo(11% vs. 0.0%). The rates of the predicted sex were female 61% vs. 56%, and male 39% vs. 44% in vivo and in vitro, respectively. The rates of survival following different biopsy methods were seen between punching and bisection method in vivo and in vitro (100% vs. 89% and 100% vs, 78% respectively). Biopsy method by punching was significantly (p<0.05) higher than bisection between produced embryos in vivo and in vitro. The present data indicate that with microblade after punching for embryo sexing results in high incidence of survivability on development after embryo biopsy. It is also suggested that LAMP-based embryo sexing suitable for field applications.

Cryopreservation induces sublethal damage to the spermatozoa, which leads to their reduced fertile life. This study was designed to determine effect of glycerol and ethylene glycol as cryoprotectant in extender on improve the freezability of Jeju horse semen. The semen was cryopreserved with glucose-EDTA extender containing each 5% glycerol, 5% ethylene glycol, 8% glycerol or 8% ethylene glycol, respectively. Post-thawed sperm were evaluated motility, viability, Membrane integrity and acrosome integrity. Post-thawed sperm motility were not significantly differences among treatments. However, sperm viability were significantly higher (p<0.05) in 8% glycerol (39.85% ±11.41) than in 5% glycerol treatment (18.08%±1.61). In membrane integrity, swelling sperm ratio was significantly higher (p<0.05) in 8% glycerol (34.12%±11.02) than other treatments. In the percentage of capacitated sperm assessed by CTC staining, F pattern was significantly higher in 8% ethylene glycol than 5% glycerol and 5% ethylene glycol (p<0.05). B pattern ratio was significantly increased in 5% ethylene glycol compared with 8% glcerol and 8% ethylene glycol (p<0.05). Moreover, 8% ethylene glycol treatment was significantly decreased AR pattern ratio compared with other treatments (p<0.05). It is concluded that treatment of 8% glycerol was improved the sperm viability and 8% ethylene glycol was improved the sperm ascrosome integrity after thawing. However, they were not significantly difference between 8% glycerol and 8% ethylene glycol on post-thawed sperm viability. Therefore, 8% ethylene glycol was more effective sperm cryoprotectant than 8% glycerol in Jeju Horse.

This study was designed to determine whether low-density lipoproteins (LDL) from egg yolk and taurine, hypotaurine and trehalose as antioxidant in extender improve the freezability and fertility of Korean Jeju Black Bull semen. The semen was cryopreserved with tris egg yolk extenders containing 7% glycerol and treated 4% LDL, 20 mM taurine, hypotaurine and trehalose. Frozen-thawed sperm were evaluated motility, viability, membrane, and acrosome integrity and sperm penetration ability. The results were compared to semen cryopreserved in tris egg yolk extender only as control. Frozen-thawed semen evaluation cleary indicated that the addition of LDL and LDL-antioxidants (taurine, hypotaurine and trehalose) combination were significantly improved (p<0.05) the viability (%; with staining test using eosin-Y) compared to control spermatozoa. Also, in membrane integrity (%; with supravital hypo-osmotic swelling test), not only LDL-antioxiants combination but also LDL were significantly increased (p<0.05) the swelled sperm using HOST compared to control. Sperm acrosome integrity state was classified by CTC (chlortetracycline) staining test. F pattern was significantly increased in LDL-antioxidant combination than control (p<0.05) and B pattern was not significantly differences among all treatments and control. However, AR pattern was significantly decreased in LDL-antioxidants combination than control (p<0.05). Pronucleus formation and sperm penetration index (SFI) were significantly increased in LDL and LDL-antioxidants combination than control (p<0.05). Especially, LDL-taurine significantly improved pronucleus fomation and SFI than LDL (p<0.05). It was concluded that LDL and LDL-antioxidants in extender improved the freezability and fertility of Korean Jeju Black bull spermatozoa.

Sperm examination is an important tool in estimating the fertilizing capacity of an ejaculate. The number of spermatozoa in a semen dose, morphology and motility are important for the fertilization process. By evaluation of semen, artificial insemination (AI) using high quality of semen can increase fertilization rate. Boar semen is subject to contamination by various pathogens that can result in fertility disorders in sows. Among these pathogens, porcine reproductive and respiratory syndrome virus (PRRSV), porcine parvovirus (PPV), porcine circovirus-2 (PCV-2) are of particular importance and accurate monitoring prior to and during the presence of boars in AI stations is essential. Because of the high risk of dissemination of disease via AI, The absolute goal is to provide pathogen-free semen and this is feasible with the adequate measures. The disease affects boars semen causes a significant reduction quality. In this study we investigated the characterization boar semen in Jeju, interaction of pathogenic virus infection with characterization of boar semen. Forty-two boar semen from 13 farms were investigated. The semen were stored during 5 days at 17℃ and the sperm qualities in the stored semen were analysed. Visual-motility assessment is a tool (Computer- Assisted Semen Analysis) used to determine the quality of boar semen. Percentage of morphologically normal spermatozoa were assessed. PRRS ,PPV and PCV-2 were detected in boar semen using PCR. The motion characteristics in boar semen was showed 68.4±9.1% for motility, 48.6±7.1 μm/s for VAP, 45.3±7.0 μm/s for VSL, 79.1±8.7 μm/s for VCL, 1.3±0.2 μm/s for ALH, 8.3±0.4 Hz for BCF, 93.6±3.5% for STR, 57.9±6.4 % for LIN. The percentage of sperm with abnormal head, midepeice and tail were 0.3±0.7%, 14.4±12.5%, 4.9±6.6%, respectively. Based on the PCR method, PPV was detected in 20 samples (48%). However, PCV-2 and PRRSV were not detected in any cases. Marked differences in motility and morphology between PPV negative and PPV positive semen were not observed. Sperm cell production was not affected by PPV infection. However, slight increases in detached head, coiled tail after infection were observed (p<0.05). The motility of semen in Jeju is similar to case comparing with other regions in Korea. Although PPV in semen was not affected in semen quality, there is the high risk of virus excretion in the semen of Jeju boars. Therefore continuous screening tests for some particular pathogens in boar semen would be warranted.