본 연구는 국내에서 생산된 주요 동계 사료작물인 청보리 및 청호밀 사일리지를 이용하여 거세 한우의 비육후기(25~30개월 령) 용으로 조제된 TMR의 소 체내 이용성을배합사료 및 볏짚으로 구성된 관행 사료와 비교하고자 반추위 누관이 장착된 소 3두를 이용하여 3 × 3 Latin squaredesign 방법으로 실시되었다. 시험동물에는 비육 후기용 배합사료 및 볏짚을 분리 급여하였거나 (대조구) 청보리 사일리지 TMR (BS-TMR) 또는 청호밀 사일리지 TMR (RSTMR)을 급여하였다. 전체 채취시간에 걸쳐 반추위액의pH, 암모니아 농도 및 총 휘발성지방산 농도는 시험사료간 차이가 없었다. Acetate (C2)의 조성 비율은 사료 급여후 1시간에서 청보리 사일리지를 섭취한 처리구 (BS-TMR)에서 낮은 (p<0.033) 값을 제외하고는 대부분의 채취시간에서 시험사료 간 차이가 없었다. Propionate (C3) 및 butyrate조성비율과 C2/C3 비율 역시 모든 채취시간에서 시험사료간 차이가 없었다. 사료 내 주요 성분의 반추위 유효분해율과 전장소화율은 전체 시험사료 간은 물론 BS-TMR과RS-TMR 간에도 차이가 없는 것으로 나타났다. 본 대사시험 결과로 미루어보아 청보리 및 청호밀로 제조된 TMR의급여 가치가 배합사료 및 볏짚의 분리급여와 차이가 없는것으로 여겨진다.

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.

This study was conducted to investigate the effect of age on the oxidative stability of Hanwoo (Korean cattle) cow beef. The samples of M. longissimus dorsi from 65 and 133 months-old-Hanwoo cows (4 and 3 heads, respectively) were stored at 4±0.2℃ for 9 days. The proximate composition, total myoglobin content, pH value, water-holding capacity and drip loss were not significantly affected by age. At day 9 of storage, the lipid oxidation (2-thiobarbituric acid reactive substances) was significantly (p<0.05) higher in beef from 133 months-old-cows than 65 months-old-cows. Lower (p<0.05) oxymyoglobin concentration and R630-R580 value and higher (p<0.05) metmyoglobin concentration were observed in beef from 133 months-old-cows at day 9 of storage than 65 months-old-cows. The CIE a*, b* and C* values were significantly (p<0.05) lowered in beef from 133 months-old-cows at day 9 of storage than 65 months-old-cows. It could be concluded that old age decreased the lipid oxidation stability and myoglobin oxidation stability in Hanwoo cow beef.

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.

The objective of this research was to investigate the effect of dietary cottonseed meal (CSM) supplementation on carcass and meat quality characteristics of Hanwoo (Korean cattle) bulls. Twenty two months-old-Hanwoo bulls of eight heads (4 heads per treatments) were fed for 5 months on a diet with or without (control) CSM (500 g/head/day). The samples of M. longissimus dorsi from carcasses were used for the quality measurement. Dietary CSM supplementation did not affect the carcass traits of bulls and pH value, drip loss, water-holding capacity and fatty acid composition of beef loin. Crude protein and crude ash content of beef loin were significantly (p<0.05) higher in beef loin from bulls fed CSM than in the control. During storage of 5 days at 4℃, CIE a*, b*, C* and total color values were significantly (p<0.05) higher in beef loin from bulls fed CSM than in the control. Overall, the present results indicate that dietary CSM supplementation may improve the oxidation stability of Hanwoo beef loin.

This study was conducted to estimate heritabilities and genetic and phenotypic correlations among growth traits (weaning weight, weight of 12 month, weight of 18 month, weight of 24 month). Growth data collected from steers (n=3,321) and 8,818 animals in pedigree were used for estimation of genetic parameters. Traits evaluated included live weight at 4 month (W4), 12 month (W12), 18 month (W18) and 24 month (W24). Parameters were estimated using an multitrait animal model and derivative-free restricted maximum likelihood procedures. Heritabilities estimated for growth traits at W4, W12, W18 and W24 were 0.21, 0.11, 0.08 and 0.14, respectively. Genetic correlations were in the range of 0.79 to 0.87. Phenotypic correlations were in the range of 0.16 to 0.81.

This study evaluated a method of sorting X and Y chromosomes based on size using the forward angle light scatter related refractive index (FSC) of a flow cytometer. Hanwoo bulls sperm were separated to X and Y chromosomes by the parameters of FSC or Hoechst 33342 intensity. As a result, using monitor program linked flow cytometry during sorting processing, the purities were or for the X-fraction and or for the Y-fraction in the two sperm sorting methods. There were no differences in the X and Y ratios (X and Y %) between the sperm sorting methods based on FSC or DNA content. The proportions of female and male embryos used for in vitro fertilization and development were or , and or when sperm were processed using the sex sorting method by FSC or Hoechst 33342. In conclusion, further study is needed to determine the optimum procedure and improve the nozzle to enhancing sorting accuracy or efficiency. Also, the findings of this study do not negate the possibility that the difference method of sperm sorting cannot use a UV laser beam.

This study evaluated a sexed sperm ability to produce embryos by flow cytometer. Hanwoo bulls sperm were separated to X and Y sperm via Hoechst 33342 stained with near UV laser or performed the pre-sorted without near UV laser beam in flow cytometry. Pre-sorted sperm had significantly higher viability (84±1.15 %, p<0.05) compared to other sorted groups in frozen-thawed semen. For fresh semen, pre-sorted sperm had the higher viability (79±3 %, p<0.05) than those of the X and Y sperm (44.7±1.67 and 41.7±1.2 %) separated by differences of DNA content. On the other hand, pre-sorted and X sperm sorted according to differences in DNA content had significantly higher viabilities (24.3±1.2 and 25.7±0.9 %, p<0.05) compared to that of the sorted Y sperm (13.7±1.2 %) in the hypoosmotic swelling test. The proportion acrosome reaction in the sorted X sperm was higher (55.0±1.7 and 45.0±1.5 %) than those of the sorted Y-sperm (32.3±0.9 %, p<0.05). However, the sperm morphologies of the sorted groups were not significantly differences. In conclusion, the sex-sorting procedure by flow cytometry affected some characteristics of Hanwoo sperm. Further study is needed to determine the optimal procedures to enhance male and female embryos and sorting accuracy.

Economic traits are quantitative traits and are mostly controlled by a large number of genes. Some these genes tend to have a large effect on quantitative traits in cattle and are known as major genes primarily located at quantitative traits loci (QTL). However, in practice, QTL is linked to allele associates of the gene controlling traits of interest. It is hypothesized that if QTL explaining a part of genetic differences between animals are detected, the effect of the genes located at QTL could assist in estimating an animal’s true genetic value. Therefore, QTL information could probably provides accuracy of breeding value estimation as well as more genetic gain through selection of animals at relatively younger age. Marker assisted selection (MAS) is the indirect selection process where a quantitative trait of economic importance is selected not just based on the trait itself but also on the basis of marker linked to QTL. MAS could be useful for traits that are difficult to measure, exhibit low heritability, and are expressed late in development. Major genes which are responsible for QTL could possibly be identified first by using different techniques such as gene expression analysis and QTL mapping. Thereafter, the information generated could be implemented for MAS in estimating breeding value. In this review we focused on delivering genome information into Hanwoo breeding program.

This study was carried out to investigate the effect of muscle part and aging period on free amino acids and aroma compounds of Hanwoo (Korean cattle) cow beef. The M. longissimus (ML) and M. semitendinosus (MS) from 101 mon-old-cows were aged at 2℃ for 14 d. The free amino acids concentration increased in both ML and MS on 14 d of aging. In ML, asparagine, glutamine, histidine, glycine, threonine, arginine, tyrosine, phenylalanine, isoleucine, leucine and lysine were significantly (p<0.05) higher than those in MS. Varieties of aldehydes, ketones, alcohols, hydrocarbons, nitrogen and sulfur compounds were detected in both ML and MS and majority of these compounds showed increasing trend on aging. The ML had higher 14 aldehydes (acetaldehyde, 2-methylpropanal, 3-methylbutanal, 2- methylbutanal, pentanal, hexanal, heptanal, E-2-heptanal, octanal, 2-octenal, 2-nonenal, E-2-decenal, E,E-2,4-decadienal and 2-undecenal), 5 ketones (2-propanone, 2,3-butanedione, 2-butanone, 2-heptanone and 2,3-octanedione), 4 alcohols (ethanol, 1-pentanol, 1-hexanol and 1-octanol), 3 hydrocarbons (3-ethyl-2-methyl-1,3-hexadiene, 3- methyldecane and 2,2-dimethyloctane) significantly (p<0.05) compared with MS. However, the MS had higher 5 nitrogen and sulfur compounds (methanethiol, dimethyldisulfide, fufural, 2,5-dimethylpyrazine and 2-octylfuran) significantly (p<0.05) compared with ML.

The Oct-4 (octamer-4), a member of the POU family transcription factor, is expressed in early mouse embryogenesis and in pluripotent embryonic stem (ES) lines. Oct-4 expression is thought to remain confined to the germline after gastrulation in the embryo. Therefore, the study was designed to, study the location of Oct-4 protein in the ovaries, placenta and testis of Korean native cattle (Hanwoo). Expression of Oct-4 mRNA in the ovaries and placenta of bovine was confirmed by RT-PCR and immunohistochemical analysis. Oct-4 was expressed in granulosa, thecal cells irrespective of the shape and size of follicles and endometerium of Korean native cattle (Hanwoo). Expression of Oct-4 was profound in all the tissues of Korean native cattle (Hanwoo) suggestung their role in them. Oct-4 localization and expression could contribute to further developmental studies in Korean native cattle (Hanwoo).

This study was performed to investigate the FSH levels for superovulation procedure in Korean Native Cattle (Hanwoo). The effectiveness of 200 mg and 400 mg of FSH to initiate superovulation was examined in Hanwoo. Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH group was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. Also, 400 mg FSH group was treated with 80, 60, 40, 20 mg FSH levels. On the 3rd day administration of FSH, 25 mg PGF2α was administered and CIDR was withdrawn. Donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 μg GnRH at time of 1st insemination and embryos were recovered 8 days after the 1st insemination. As a results, average number of CL treated with FSH 200 mg was higher as 20.9±1.20 than 15.8±0.63 for donors treated with FSH 400 mg, respectively(p<0.05). Treated group of 200 mg FSH level increased (p<0.05) the number of embryos recovered per procedure compared to 400 mg FSH level (18.2±1.18 vs. 12.38±0.52, respectively). When treatment of 200 mg FSH was performed, average transferable embryos/ova increased (p<0.05) to 14.1±1.12 from 6.8±0.33 of treated of 400 mg FSH. Group of 200 mg FSH increased (p<0.05) to 8.3±0.76 from 2.0±0.26 in morula stage compare to 400 mg FSH group. Mean of total early blastocyst and expanded blastocyst stage embryos was similar (p<0.05) between the 200 mg and 400 mg FSH levels group (4.7±1.19 vs. 2.9±0.18 and 1.2±0.40 vs. 1.9±0.17). These results suggest that 200 mg FSH level-based superovulation protocol with CIDR may be effectively used for production of superior embryos in Hanwoo. In other words, the less level of FSH may be effectively applied for Hanwoo (Korean Native Cattle), because Hanwoo was smaller body size than beef or daily cow.

포유동물 수정란의 동결보존기술은 최근 기후 변화에 따른 생물종 다양성을 보존하기 위해서 중요하게 여겨지는 연구 분야이다. 따라서 멸실 위험에 처한 동물의 개량과 증식, 보존과 복원 및 생명공학의 분야에 이르기까지 응용 기술은 다양하게 이용되어진다. 본 연구에서는 한우 수정란의 동결 후 생존성 향상을 위해서 동결 방법에 따른 체내 외수정한의 내동성을 조사하였다. 완만동결에 따른 체내 외수정란의 동결 융해 후 수정란의 재확장률은 89.6%와 81.5% 그리

Spermatozoa sorted by flow cytometry have been successfully used to produce offspring in domestic animals and are commercially available for cattle. Also sheath fluid is the important environment for viability of sex-sorted sperm in flow cytometry. The aim of this study was to investigate whether or not HEPES (N-2-hydroxyethylpiperazine-N'-2-Ethanesulfonic acid) has any effect on the viability in sex-sorted Hanwoo (Korean native cattle) sperm. In this study, the semen was collected from Hanwoo of Hoengseong Livestock Cooperation by artificial vagina method then pooled and subjected to cryopreservation in straws. Sperm were cultured for 0, 30, 60 and 120 min with 0, 2.5, 5, 7.5 and 10 mM of HEPES added to the sheath fluid and incubated at 4, 20 and 38, respectively. For the cytometric analysis the frozen-thawed semen was extended with 5 mM HEPES extender to final concentration ( spermatozoa) at 4, 20 and 37. Sperm viability was assessed with SYBR-14 and propidium iodide (PI) staining. This study shows that the viability of sperm was decreased with prolongation of incubation time in all of test. But the viability of sperm which were treated with 38 was gently decreased than that of treated with other temperature. The viability of the control was sharply decreased (p<0.05) than all of the HEPES treatment group at 60 to 120 min in 38. X-sexed sperm was more sensitive than Y-sexed sperm to temperature during f10w cytometry (p<0.05). In conclusion, the results of this study suggest that the sheath fluid with 5 mM HEPES has effect on maintenance of viability after sperm sexing at 37 in Hanwoo.

This study was performed in order to determine optimum flushing solution using the direct embryo collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3 day administration of FSH, 25 mg was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 GnRH at time of 1 insemination and embryos were recovered 8 days after the 1 insemination. Embryo collection from superovulated donors were performed to flushing by DEC and conventional method. As a results, the average number of recovered embryos were significantly higher as 19.11.40 with DEC method than 12.00.44 with conventional embryo collection method, respectively (p<0.05). Also, The average number of transferable embryos were significantly higher (p<0.05) as 15.81.72 with DEC method than 6.90.35 from conventional embryo recovery procedures. Meanwhile, number of recovered embryos and number of recovered transferable embryos following the number of flushing times until 6 flushing were significantly higher as 8.60.53 and 8.60.53 from 2 flushing time than other groups (p<0.05). No. of Ear. B stage embryos were significantly higher as 3.90.90 and 3.90.90 with 2 flushing time in total collected embryos and transferable embryos (p<0.05). Com M stage embryos were significantly higher as 3.71.00 in 2 flushing time and as 2.20.76 in 3 flushing time for recovered embryos (p<0.05). In transferable embryos, Com. M stage embryos were significantly higher (p<0.05) as 3.71.00 in 2 flushing time and as 2.20.76 in 34 flushing time, also. No. of degradation embryos was significantly higher as 2.20.72 in 5 flushing time, On the other hand, degradation embryos was not observed in transferable embryos (p<0.05). In conclusion, these results suggest that DEC method should effective methods for production of in vivo embryos using less flushing solution following perform until 4 flushing time than conventional embryo collecting method. Also, it might be effectively collection of transferable embryos following more less procedure times compared to conventional embryo recovery methods.

This study was performed in order to simplify the operation and minimize stress of donor and be readily available in the field with low cost and high quality embryos using the Direct Embryo Collection (DEC). Donors, at random stages of the estrous cycle, received a CIDR. 7 days later, 200 mg FSH was treated with 40, 30, 20, 10 mg FSH levels in declining doses twice daily by intramuscular injection for 4 days. On the 3rd day administration of FSH, 25 mg was administered and CIDR was withdrawn. After FSH injections were complete, donors were artificially inseminated twice at 12 hr intervals. The donor cattle received 250 GnRH at time of 1 st insemination and embryos were recovered 8 days after the 1st insemination. Embryo collection from superovulated donors was performed to flushing by non-surgical methods of 3-way, 2-way and DEC (l-way). The average number of recovered embryos were 11.250.63, 12.50.65 and 11.750.48 from operations of 3-way, 2-way and DEC methods, respectively. There were no significant differences among the embryo collection methods. Also, The average number of transferable embryos were 6.250.48, 7.250.48 and 7.250.63 from each embryo collection procedures. The number of transferable embryos was no differences among the 3-way, 2-way and DEC methods, respectively. Meanwhile, the ratio of transferable embryos for all recovered embryos from DEC methods was higher as 61.7 % than 55.6 %, 58 % from methods of 3-way, 2-way. And the flushing solution required for recovering embryos by DEC method was significantly lower as 0.280.32 1 than 1.80.12 1, 1.750.10 1 from 3-way, 2-way methods (p<0.05). Also, the time required for recovering embryos by DEC methods was significantly lower as 272 min than 513, 452 min, respectively (p<0.05). In conclusion, these results suggest that DEC method for embryo collection may be effectively used for production of in vivo embryos using less flushing solution and, it might be effectively available in the field compared to conventional embryo recovery methods using 3-way or 2-way balloon catheter.

In general, zona pellucida (ZP) of the blastocyst has to be removed first, then either isolated the inner cell mass (ICM) or ZP-removed whole blastocyst, which is then cultured on the feeder layer to induce ICM outgrowth for the generation of embryonic stem cells (ESC). However, it is unclear whether ICM isolation before seeding on feeder layer is beneficial or not because the interaction between ICM and trophoblasts may affect cellular growth and/or pluripotency during the culture on the feeder. In the present study, two ZP removal methods (mechanically by splitting with a 28-gauge needle versus chemically by the treatment of acid-Tyrode's solution) and two ICM isolation methods (ZP-free whole blastocyst seeding versus mechanical isolation of ICM) were evaluated for the efficient isolation and culture of putative parthenogenetic bovine ESC. The number of maintained outgrown colonies was counted in each experimental group. As the result, mechanical removal of ZP with a needle and followed by whole ZP-free blastocyst seeding on feeder cells tended to attach more on the feeder layer and resulted in more outgrown colonies with its simple and less time-costing benefits. Currently we are generating ESC lines in HanWoo cattle by using this method for initial outgrowth of the parthenogenetic bovine blastocysts.

This study was carried out to investigate the effect of amino acids complex and choline supplementation on the antioxidant enzyme activities and oxidative stability of Hanwoo (Korean cattle) beef. Fifteen months-old-Hanwoo steers were assigned into two groups and fed on a basal diets with or without amino acids complex (≥25% L-lysine monohydrochloride+≥8% DL- methionine)+choline (≥25% choline chloride) for 12 months. After slaughter, the M. longissimus from carcasses were stored at 4℃ for 7 days. Catalase, glutathione peroxidase and total superoxide dismutase activities were found to be unaffected by supplementation of amino acids+choline. After 2 days of storage, significant decline (p<0.05) in lipid oxidation (2-thiobarbituric acid reactive substances, TBARS) was observed when supplemented with amino acids+choline. However, supplementation of amino acids+choline maintained meat color as indicated by higher CIE L* (Lightness), a* (Redness), b* (Yellowness) and C* (Chroma) values during storage. It was therefore concluded that supplemental amino acids+choline could stabilize the lipid oxidation stability and meat color in Hanwoo beef.