The aim of this study was to investigate the effects of different rearing spaces on growth performances, behavioral traits, and blood parameters in Hanwoo (Korean Native Cattle) heifers and steers. The average daily gain and feed conversion ratio were measured to assess the growth performance. A total of 54 Hanwoo heifers and steers at 18 months old, were housed separately in groups of 2, 3 or 4 heads in a pen (32 m2) for nine months. In the case of the heifers, the growth performances were significantly (p<0.05) improved by increasing the 2 head group size with the exception of the 18 month old heifers. However, among the steers the growth performances were significantly enhanced (p<0.05) after 18 months of age, by increasing the 3 heads group size. In addition, the heifers in groups of 2 heads spent less time standing and more time lying down or walking, and also had a significantly (p<0.05) lower frequency of self-grooming, scratching, and fighting. These findings are similar to the results obtained for steers housed in 2 or 3 heads per group. The steers housed in 2, 3 heads groups spent more time feeding and consequently had a significantly (p<0.05) lower frequency of pair-wise grooming and Leaning. The heifers and steers housed in groups of 4 heads showed lowest red blood cell counts, but highest cortisol content with statistical significance at p<0.05. The heifers in groups of 2 heads had the highest hemoglobin concentrations (p<0.05). On the other hand, white blood cell count was found to be the lowest in groups of 2 steers with statistical significance at p<0.05. We conclude that a large rearing space improved growth performances, behavioral characteristics, and blood parameters in Hanwoo heifers and steers. In the future, this data could be used to improve the welfare of rearing Hanwoo cattle.

Bovine papillomaviruses (BPVs) are DNA oncogenic viruses that belong to the Papillomavirus genus. The purposes of the present study were to histopathologically diagnose skin warts on Korean native cattle from Jeju Island and to detect the presence of BPVs from tissue samples of the skin warts, using the polymerase chain reaction (PCR) assay. The sites of 10 skin wart involvement were the head (40%: 10% were in both the head and neck), shoulder (30%), neck (20%), and tail (20%), including the perineum. All skin warts were confirmed as fibropapillomas, as characterized by the abundant proliferating fibrous tissue in the dermis and increased epithelial covering of variable thickness. According to the PCR analysis, five out of 10 (50%) cattle were positive for BPV-1, and six (60%) cattle were positive for BPV-2. Two cattle were positive for both BPV-1 and BPV-2. To the best of our knowledge, this is the first study of BPV-1 and BPV-2 infections of skin warts from Korean native cattle in Korea.

Calculating break-even price of calf production is closely associated with reproductive efficiency. To determine the price, we need data from reproduction records including number of claves weaned, number of cows exposed for breeding, and annual cash coast per cow, and average weaning or market weight of claves sold and retained. Unfortunately, the data were not available in Korea native cow (Hanwoo). To evaluate the performance and the price, we collected calving interval from about 60,000 cows for last 10 years and estimated reproductive performance. Calving interval was increased 4.3% and pregnancy rate was decreased about 1.4∼2.8% year-on-year. Increases in growth rates of number of cow and semen per calf supported the low reproductive performance. Finally, break-even price was calculated using estimated percent calf crop and demonstrated that growth rate of break-even price is larger than that of annual cash per cow, suggesting cow-calf profitability and financial efficiency in Korea native cow (Hanwoo) is getting worse.

The objective of this study was to monitor health conditions of genetically identical somatic cells cloned Korean white cattle, endangered indigenous cattle (EIC) and indigenous cattle (IC) by analysis of hematologic characteristics. Naturally ovulated oocytes and donor cells were used for somatic cell nuclear transfer (SCNT). Donor cells and enucleated oocytes were followed by electric fusion, chemical activation and surgical embryo transfer into the oviducts of surrogate females. Two recipients became pregnant; two maintained pregnancy to term, and one live cattle were delivered by caesarean section. The cloned Korean white cattle were genetically identical to the nuclear donor cattle. As a result, the mean values of RBC and platelet of cloned cattle and white cattle were significantly decreased by age (P<0.05). The mean values of RBC, HCT, MCV and MCHC between cloned cattle and IC of the same age (1∼2 years) showed the statistical significance (P<0.05). Also, in the WBC of Korean white cattle, the estimated values were decreased according to the age from 12.0×103/μl under 1 year to 11.0×103/μl over 1 years respectively. Although clone-cattle had lower numbers of RBC than reference range, the most of RBC and WBC related heamatologic results of cloned cattle were not different when compared to reference range. This study suggests that cloned Korean white cattle derived from SCNT did not have remarkable health problems, at least in the growth pattern and hematological parameters. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

These studies were conducted to establish the practical Hanwoo (Korean native cattle) improvement system through the combining of embryo transfer technology and confirming individual Hanwoo oocyte culture system and to investigate that correlation of Hanwoo carcass classification (intramuscular marbling) and oocyte donor for blastocyst production in vitro. In case of Hanwoo, the carcass meat quality grades were divided to grade 1⁺⁺, 1⁺, 1, 2, and 3 depends on fat distribution of longest muscle cross-sectional surface. As results, the numbers of follicular oocytes collected from individual fundamentally-registered Hanwoo yielded 1⁺⁺, 1⁺, 1, 2 and 3 meat quality were 9.5, 9.4, 8.5, 8.8 and 8.8 per ovary, respectively. The numbers of retrieval oocyte from follicles were significantly higher in the cattle yield 1⁺⁺, 1⁺ meat quality than in the cattle yield 1, 2 and 3 meat quality (p<0.05). The rates of blastocyst formation were 18.2, 21.3, 29.4, 30.9, and 31.5% in the cattle yield 1⁺⁺, 1⁺, 1, 2 and 3 meat quality of after in vitro maturation, respectively. It was significantly lower in the cattle yield 1⁺⁺ and 1⁺ meat quality than in the cattle yield 1, 2 and 3 meat quality (p<0.05). In order to evaluate embryos quality, TUNNEL assay was conducted for each meat quality grade using blastocyst stage embryo on day 8. The results showed that apoptosis cell number was higher tendency in the cattle yield 1⁺⁺and 1⁺ meat quality (81 and 79, respectively) than in the cattle yield 1, 2 and 3 meat quality (51, 48 and 50, respectively) but there was no statistical significance in each group. After embryo transfer, the conception rate of recipient was 53.5 (23 out of 43), 52.1 (38 out of 73) and 58.0% (58 out of 100) in the meat quality of 1, 1+ and 1++, respectively. These results showed that the conception rate was significantly higher in the cattle yield 1 meat quality than in the cattle yield 1⁺⁺, 1⁺ , 2, and 3 meat quality (p<0.05). In summary, these results indicate that the application of confirming Hanwoo individual oocyte culture system and embryo transfer technology can make good use of the genetic resources conservation and improvement of Hanwoo. Relevance of the meat quality grade and reproductive ability of carcasses of Hanwoo will be considered to be one of the effective means for the associated research with obesity and reproduction.

The purpose of this study is to produce wanted sex progeny of genetically confined White Hanwoo (albinism) with preselected sex sperm. One bull of White Hanwoo was chosen for semen donor and X sperm was sorted by MoFlo XDP cell sorter. To compare the pregnancy and birth rates, KPN straw was used as control, total number of unsorted sperm was 20×106/straw. Sexed X frozen semen with 2×106 cells or 4×106 cells per straw were in seminated twice on Hanwoo heifers. The abnormality of the sexed X semen was 24.9 ± 7.31% and distal reflex abnormality of mid piece was significantly (p<0.05) higher (11.7%) compared with that of KPN 768 (5.6%). There were no differences on the pregnancy and birth rates between 2×106 cells or 4×106 cells of X-sperm but KPN semen showed significant differences (p<0.05). The pregnancy rates of KPN 768, 2×106 cells and 4×106 cells X-sperm of White Hanwoo cattle were 85.0%, 26.3% and 50%. The birth rates were 80.0%, 15.8% and 21.4%, respectively. The female offspring rates of KPN 768, 2×106 cells and 4×106 cells X-sperm of White Hanwoo cattle were 43.8%, 100% and 100% (p<0.05). These results indicated that sex sorted White Hanwoo could be used for the production of wanted progeny with 2×106 cells/straw for AI. To increase the efficiency of calf production, the sperm number of sex sorted semen will be optimized for sex selection of White Hanwoo progeny.

The importance of genetic resource preservation has been highlighted in the literature as a means of maintaining genetic diversity. Investigations for hematologic values and the differential count of white blood cell count (WBC) for Korean indigenous cattle (KIC) and endangered indigenous cattle (EIC) are rarely performed. Therefore, the objective of this study was to investigate the hematologic values of total 40 EIC (White, Black, Mini cattle) and 35 KIC as control by analysis of hematologic characteristics. As a result, the mean values of RBC and platelet of EIC were significantly decreased by age (p<0.05). The mean values of RBC, HCT, MCV and MCHC between EIC and KIC of the same age (2 ～3 years) showed the statistical significance (p<0.05). Also, in the WBC of EIC, the mean values were decreased according to the age from 13.9×103/μL～12.7×103/μL under 1 year to 9.1×103/μL～11.5×103/μL over 2 years respectively. In the differential count of WBC of EIC (White, Black, Mini cattle), it showed generally the rates of 40.2%, 52.2%, 49.0% lymphocyte and 27.2%, 33.9%, 32.0% segmented neutrophil from 2～3 years respectively. Result of this study will be used for establishing reference range for blood analysis in EIC such as white, black and mini cattle. This study reported hematological values which could serve as baseline information for comparison in conditions of nutrient deficiency, physiological and health status of endangered Korean native cattle. In addition, this study provides a valuable resource for further investigations of the preservation of rare genetic stocks underlying traits of interest in cattle.

Cattle breeds were classified previously into three different haplogroups (Y1 and/or Y2 in Bos taurus and Y3 in B. indicus) based on Y chromosome-specific polymorphisms. In particular, a rapid and unambiguous classification method was reported recently. However, a haplogroup classification of Korean native cattle breeds has not been reported. In this study, 196 animal samples from four Korean native cattle breeds (Hanwoo, Chikso, Heugu, and Jeju black cattle) and six exotic breeds were used to determine the Y chromosome-specific haplogroup classification. We amplified an 81 bp indel region within intron 26 of the USP9Y gene and performed electrophoresis to classify the Y1 and Y2 haplogroups. Moreover, enzyme digestion was carried out with the SspI restriction enzyme to classify the Y2 and Y3 haplogroups. Finally, sequence variation in each haplogroup was confirmed by DNA sequencing. All animals in the four Korean native cattle and two exotic breeds (Charolais and Simmental) belonged to the Y2 haplogroup. Three other exotic breeds (Holstein, Angus, and Hereford) belonged to Y1 haplogroup. Japanese black cattle were divided into both the Y1 and Y2 haplogroups. The Y3 haplogroup corresponding to B. indicus was not found in this study. In conclusion, Korean native cattle breeds originated from B. taurus without introduction from B. indicus. In addition, they showed the same paternal heredity pattern which belonged to only Y2 haplogroup. These results can be used to investigate the origin of Korean native cattle breeds.

2013년 11월부터 2014년 7월까지 가축유전자원시험장 장내에서 사육 중인 한우 및 칡소 206두(한우 187두, 칡소 19두)를 대상으로 연령별, 종별 그리고 성별로 각각 분류하고, 이들에 대한 혈액학적 수치를 확인하였다. 혈액학적 검사 결과에서 1년 이하에서 5년 이상으로 연령이 증가됨에 따라 RBC(12.0~9.5x106/μl로)와 PLT치(717~432x103/μl로)에서 각각 유의성 있는 감소(P＜0.05)가 인정되었다. 동일 연령(2~3년)의 한우군과 칡소군 간에서의 비교에서는 RBC(10.1~10.4x106/μl)와 HCT(44.8~42.1%), MCV(49.4~44.9 fl) 및 MCHC (28.9~31.1 g/dl)에서 두 품종 간의 차이점이 인정되었으며, MCV와 MCHC에서도 통계적 유의성이 인정되었다(P＜0.05). 또한, 백혈구계 검사 결과에서도 연령이 증가함에 따라 WBC 수치(1년 이하의 13.8에서 5년 이상의 9.5x103/μl로)가 감소하는 경향이 인정되었으며, 백혈구 분포의 백분율에서는 전반적으로 림프구가 46.2%를, 분엽형 호중구가 35.6%로 나타났다. 한우군과 칡소군 간의 백혈구계 비교에서는 칡소군의 분엽형 호중구치가 2.7x103/μl로서 한우군에 비하여 유의성 있는 저하(P＜0.05)가 인정되었으며, 림프구치는 5.2x103/μl로 4.7x103/μl 한우군에 비하여 높게 나타났다. 임신우군과 비임신우군 간의 비교 분석에서는 Hg(9.9 g/dl)과 HCT(40.5%)이 비임신군에 비하여 통계적 유의성이 있는 저하(P＜0.05)가 인정되었으며, 총 백혈구수와 분엽형 호중구수, 림프구수 모두 비임신우군에 비하여 높게 나타났지만, 통계적 유의성은 인정되지 않았다.

Sexed semen is commonly used for the production of calves of the desired gender. Gender selection is important in animal production industries. For example, female cattle are required for the dairy industry while males are preferred in the beef cattle industry. The present study was to assess the in vivo embryo production efficiency using the semen separated according to sex during superovulation in Hanwoo. Seventy Hanwoo donor cows were flushed on day 7 of estrus cycle with same FSH and artificial insemination by the same technicians. Embryos were recovered on 7 days after the third insemination by flushing the uterus with embryo collection medium. KPN semen straws used artificial insemination contained 20 million sperm (total number 60 million per donor). Sex-sorted semen straws contained 4 million sperm (total number 12 million per donor). The results obtained were as follows: No differences were observed in the efficiency of superovulation rates on KPN semen 87%, and sexed semen 100%, respectively. The mean numbers of total embryos are each 12.58±8.31 and 13.25±7.86. The mean numbers of transferable embryos, sexed semen were significantly lower than KPN semen (3.75±1.98 vs. 8.23±6.07, P＜0.05). The rates of unfertilized embryos from superovulation using sexed semen were significantly higher than KPN semen (50% vs. 15%, P＜0.05). The rate of degenerated 2-cell embryos from sexed and KPN semen was 60.87% and 11.11%, respectively (p＜0.05). In conclusion, these results indicate that superovulation using sexed semen was useful, but efficient embryo production was important to reducing the damage caused by the Flowcytometer-based sperm sorting procedure.

The objective of this study was to evaluate the characteristics of Korean Native Cattle sperm frozen-thawed with L-cysteine and/or catalase. The semen from bulls was collected by the artificial vagina method, and Triladyl containing 20% egg-yolk and/or L-cysteine (L), catalase (C) and L-cysteine + catalase was added to the diluted semen for cryopreservation. The results showed that sperm viability was significantly higher in the L-cysteine + catalase (69.49 ± 3.16%) group than in the control (60.5 ± 3.94%) group (p<0.05). Acrosome damage was significantly lower in the L-cysteine (17.12 ± 1.08%) group than in the control (21.46 ± 1.14%), catalase (20.54 ± 0.76%), and L-cysteine + catalase (19.29 ± 0.65%) groups (p<0.05). In addition, the level of intact mitochondria in the spermatozoa was significantly higher in the L-cysteine (58.65 ± 1.39%) group than in the control (50.63 ± 2.37%) group (p<0.05). The hydrogen peroxide level in the frozen-thawed sperm was significantly lower in the L-cysteine (3.74 ± 1.66%), catalase (4.65 ± 1.87%), and L-cysteine + catalase (8.11 ± 2.15%) groups than in the control (13.22 ± 1.6%) group (p<0.05). The glutathione level was significantly higher in the L-cysteine (1.33 ± 0.03%) group than in the control (1.08 ± 0.06 %), catalase (1.05 ± 0.02%) and L-cysteine + catalase (1.11 ± 0.03%) groups (p<0.05). In conclusion, L-cysteine and catalase could protect the membrane of Korean Native Cattle sperm from damage during sperm cryopreservation. Especially, L-cysteine was more effective for keeping acrosomes and mitochondria intactness during sperm cryopreservation.

Cell-free fetal RNA has been highlighted as useful tools for the fetal sex determination or other genetic inherent disorder. However, there is no knowledge about the sex determination using cell free fetal RNA in bovine field. Thus, the present study aimed to evaluate the presence of transcripts of DDX3Y, USP9Y and ZRSR2Y genes in maternal plasma of pregnant cows to determine the sex of the fetus using real-time quantitative polymerase chain reaction assay, and verify its accuracy, sensitivity and specificity compared with the molecular testing and the calf sex at birth. Transcripts of USP9Y and DDX3Y genes were expressed in the all plasma of males and females both the control group and the experimental group. However, ZRSR2Y gene was matched up with the molecular testing and the true sex in control group and has an overall accuracy of 82.6%, a sensitivity of 75%, and a specificity of 100% in experimental group. Therefore, these results indicated that real time PCR technique, as a noninvasive and cost-efficient method, is possible to determination fetal sex in the bovine species using circulating cell free RNA in maternal plasma and especially ZRSR2Y gene could be a good candidate for the RNA based sex determination work.

The aim of this study was to identify allele variability and frequencies of six microsatellite markers (BM861, INRA124, INRA189, UMN0103, UMN0307 and UMN0504) specific to the Y-chromosome. DNA samples from 147 males of three Korean native and seven exotic cattle breeds were tested. Three (Chikso, Heugu, and Jeju black cattle) Korean native cattle (KNC) breeds showed Bos taurus genotype. In the neighbor-joining tree, based on Nei’s DA genetic distance, ten breeds represented main group. In addition, Bayesian clustering result showed that 3 main clusters are for individual allele variability and frequencies. Moreover, KNC breeds showed differences in genotype with B. taurus type when compared to previous studies. The molecular information of paternal lineage in this study would be useful for the conservation and utilization of three KNC breeds as genetic resources.

The role of Fatty acid binding protein 4 (FABP4) gene is critical for lipid metabolism and for maintaining homeostasis in adipocytes. Association between Hanwoo carcass traits and FABP4 gene g.7516G>C SNP has been reported previously, however, its association and how does it influence Korean brindle and black cattle has not been demonstrated and established till date. For this purpose, the study was planned to analyze the SNP association (Single Nucleotide Polymorphism) (g.7516G>C) in FABP4 gene and gather genetic information on economic traits of Korean brindle cattle and black cattle. As per sequence of bovine FABP4 gene available (Genbank accession No. NC_007312.4), one pair of primers (5’-ATA TAG TCC ATA GGG TGG CAA AGA-3’ and 5’-AAC CTC TCT TTG AAT TCT CCA TTC T-3’) was designed to amplify a 452bp product of the FABP4 gene including the region of 7417–7868. The SNP, detected by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) methods using restriction enzyme MspA1I, was genotyped in 117 animals of brindle cattle and 24 animals of black cattle population. Statistical analysis revealed that the FABP4 genotype significantly (p<0.05) affect with carcass weight, but there was no significant association with any other economic traits was observed in brindle as well as in black cattle. In conclusion, these results suggested that SNP (g.7516G>C), located in FABP4 gene, could be used as important DNA marker of economic traits in Korean brindle cattle. Furthermore, we suggest that additional samples needs to further analysed to make related data exclusively authentic.

The melanocortin receptor type 4 (MC4R) gene is expressed in the hypothalamus and regulates energy intake and body weight. Recently, it has been reported that obesity and energy balance in human were also regulated by the MC4R gene. Therefore the objective of this study was to identify the polymorphism on the MC4R gene SNP C1786T and its association with economic traits in Korean native cattle (brindle and black cattle) by PCR-RFLP. A total of 125 cattle from the two breeds were tested for economic traits (meat quality index, backfat, thickness, carcass weight, longissimus muscle area and marbling score) and data was analyzed using SAS program. In the results, C allele had highest frequency than G allele frequency in the both breeds and the gene was significantly associated with meat quantity index and backfat thickness in brindle cattle breed. However, in black cattle, the gene was significantly associated with longissimus muscle area (p<0.05). These results suggest that C1786T SNP of the MC4R gene may be useful as a genetic marker for economic traits in the brindle and black cattle.

The objective of this study was to evaluate several types of uterine bacteria in Hanwoo. uterine bacteria from randomly selected 5 uterus was collected by flushing methods into a sterilized 1.5 ml centrifuge tube and was inoculated onto MacConkey agar and blood agar, respectively. After being incubated for 5% CO2, aerobic or anaerobic condition at 37℃ during 48h, bacterial colonies were selected and re-inoculated onto blood agar plates. Re-cultured colonies were identified by Gram staining and finally identified using Vitek system. The identified bacteria were Staphylococcus lentus, Staphylococcus sciuri, Staphylococcus vitulinus, Staphylococcus warneri of Gram (+) and Rhizobium radiobacter, Sphingomonas paucimobilis of Gram () bacteria. Although, pathogenicity of identified bacteria was unclear, the bacteria can have an effect on the uterine microenvironment. Therefore, repetitive research will be required to determine the effects of bacteria in cattle exposed to a various environment.

Solution of glycerol, ethylene glycol, sucrose, dextrose (GESD) and cryotop methods were carried out to investigate the survivability on vitrification of embryos. Embryos cultured in vitro were vitrified by GESD of 10 or 8 step and cryotop methods of 6 step, from cryopreservation step to frozen-thawed and culture step. Survival rate and ICM, TE cells of embryos were investigated after frozen-thawed 24 h. As a results, cryotop method was significantly (p<0.05) higher ( vs. , ) than GESD 10 or 8 step methods on survivability. Also, In ICM cell number, cryotop method was significantly (p<0.05) higher to cells than GESD 8 step method. TE cell number was significantly (p<0.05) highest to cells in cryotop method. On the other hand, survival rate, TE and total cell number were all the significantly (p<0.05) high, except ICM in GESD 10 step method between GESD 10 step method and GESD 8 step method. In conclusion cryotop method was to be most effective, but it is considered necessary to study vitrification method for step-by-step freezing and thawing process.

This study was carried out to effects of ethylene glycol concentration, sucrose and culture day of in vitro production embryo on slow-down freezing in Hanwoo. 6, 7, 8 and 9 day embryos produced in vitro were frozen using 1.8M EG+0.1M sucrose, 1.8M EG+0.5% BSA and 1.5M EG+0.1M sucrose media. Survivability was confirmed after frozen-thawed 24 and 48h and ICM, TE cell number were counted by Hoechst 33342 and PI staining after frozen-thawed 24h. As a result, 1.8M EG+0.1M sucrose group was most significantly (p<0.05) higher compared with the other treatment groups on survivability, TE and total cell number after frozen-thawed 24h (, and ). ICM number did not found significant (p<0.05) differences between the three treatment groups. in 6, 7, 8 and 9 day of embryos using three types of freezing media, frozen-thawed, 1.8M EG+0.1M sucrose groups with embryos cultured 8 day was significantly (p<0.05) highest survivability to after frozen-thawed 24h. 1.5M EG+0.1 sucrose group with embryos cultured 9 day was significantly higher survivability than group of embryos cultured 8 day after frozen-thawed 24 and 48h. In conclusion, 1.8M EG+0.1M sucrose media is considered to be effective to cryopreservation of embryos cultured 8 and 9 day.

After spermatogenesis, spermatozoa come in contact with fluids in the epididymis where they mature. During ejaculation, spermatozoa are mixed with secretions from prostate gland, vesicular glands, and bulbourethral glands. During natural mating, seminal plasma is deposited in the female reproductive tract eliciting various physiological and immunological responses. With the advances in proteomics, the components of seminal plasma have been identified and the information may be valuable in identifying markers for fertility. Components of seminal plasma that affect fertility have been discovered and the mechanism of action of these factors has been determined. The objective of this study was to determine the specific seminal plasma proteins from Korean native cattle, Hanwoo, and Korean native brindle cattle (KNBC) with the long term goal of improving fertilization rate. After SDS-PAGE and 2-dimensional gel electrophoresis, proteins were identified by Q -ToF analysis. They include plasma serine protease inhibitor precursor and platelet-activating factor acetylhydrolase after SDS-PAGE. Number and density of the spots in 2-dimensional gels were higher in KNBC than Hanwoo. Proteins identified from the paired spots of both breeds include chain A, bull seminal plasma PDC-109 Fibronectin Type II module, BSP-30 kDa precursor, and Spermadhesin Z13 or its precursor. Interestingly, some proteins were identified from multiple spots. The functional differences of these diverse forms of the proteins may require further studies. With their previously reported roles in sperm capacitation by these proteins, the studies on the mechanism of action, ligand interaction and the variation in the genome may help improving fertility in cattle.