This study evaluated the effect of reduced glutathione (GSH) for the reduction of stress and inflammatory response in calves inoculated with foot-and-mouth disease (FMD) vaccine. Twenty-five calves were divided into five groups of 5 calves. The negative control (NC) did not receive any vaccination or drug treatment. The positive control (PC), GSH-25, GSH-50 and GSH-100 were intramuscularly injected with GSH at concentrations of 0, 25, 50 and 100 mg / 10 kg body weight (BW), respectively, for 3 days after FMD vaccination. On day 3, 5 and 7 post-treatment, the serum cortisol and tumor necrosis factor- α (TNF-α) levels in GSH-50 and GSH-100 were significantly decreased compared with those in PC (p < 0.05). However, there was no significant difference in the serum cortisol and TNF-α levels between GSH-100 and NC 3 and 5 days post-treatment, and between GSH-50, GSH-100, and NC 7 days post-treatment. The results from this study suggest that treatment of 50 mg / 10 kg BW GSH for 3 days is useful for the reduction of stress and inflammatory response caused by FMD vaccination in calves.

한우 송아지의 생후 환경스트레스에 대한 면역반응성 및 신바이오틱제제의 급여효과를 구명하기 위해서 출생 후부터 각 처리구별로 5두씩 배치하여 총10두(2처리×5두)를 대상으로 대조구(신바이오틱제제미투여)와 처리구(신바이오틱제제 투여구)로 배치하여 총 65일간(포유기간) 시험하였다. 송아지 체중은 대조구(21.4±2.51 kg - 57.0±12.83 kg)에 비해 신바이오틱제제 급여 처리구(22.0±2.12 kg - 53.2±4.32 kg) 개체간의 체중의 편차를 줄이는 역할을 하는 것으로 나타났다. Nitric oxide는 신바이오틱제제 처리구(12.52 μM/L-39.72 μM/L)가 대조구(19.68 μM/L-64.80uM/L)보다는 낮았으며, 코티졸 농도는 한우 송아지 신바이오틱제제 처리구에서 7일령, 45일령, 그리고 65일령에서 유의적으로 감소하였고(P<0.05), 혈중 Glutathione는 65일령에서 유의적으로 증가하였다(P<0.05). SOD의 농도는 신바이오틱제제 처리구의 28일령과 65일령에서 유의적으로 감소하였고(P<0.05), H2O2의 농도는 신바이오틱제제 처리구 45일령과 65일령에서 유의적으로 감소하였다(P<0.05). Interlukin-1β 농도는 대조구에서 28일령(43.26±4.40 pg/mL)과 처리구 21일령(24.68±3.20 pg/mL)에 가장 높았으며, 14일령, 28일령, 65일령의 신바이오틱제제 처리구에서는 유의적으로 낮았다. IL-2 농도는 대조구 28일령 (218±16.94 pg/mL)과 신바이오틱제제 처리구 7일령(174.60±11.60 pg/m)에서 각각 가장 높았고, 21일령 이후 처리구에서 유의적으로 감소하였다. IL-6 농도 또한 대조구 28일령(403.20±48.19 pg/mL)과, 신바이오틱제제 처리구 65일령( 238.20±15.63 pg/mL)에서 가장 높았으며, 21일령 이후 신바이오틱제제 처리구에서 유의적으로 낮았다. PGE2 농도는 대조구 45일령(3660±463.25 pg/mL)과 신바이오틱제제 처리구 45일령(1070±141.92 pg/mL)에서 각각 가장 높았으며, 14일령 이후 신바이오틱 제제 처리구에서 유의적으로 낮았다. 앞의 결과를 종합해보면, 신바이오틱제제의 경구투여로 생체내의 싸이토카인의 체내 균형조절 및 면역 염증반응을 억제하는 데 관여하는 것으로 보인다. 그리고 신바이오틱제제의 급여로 분만 직후 한우 송아지의 체내의 변화가 체외의 변화로 이어져 가축의 성장촉진 및 열악한 주변환경 극복 등에 보다 더 확실한 효과를 기대하기 위해서는 신바이오틱제제의 급여기간을 늘릴 필요가 있을 것으로 사료된다.

The increase in the total number of cows on farms, which breed Korean Native Cattle (KNC), is associated with many problems. In particular, the services per conception and calving interval have increased. In this study, we examined the effect of dietary supplementation with vitamin and mineral complex on the estrus rate, pregnancy rate, gestation length, and birth weight of KNC calves. Multiparous or primiparous KNC were divided into 3 groups with 40 heads per group. Experimental group 1 was administered a consisting of 35,000 IU vitamin A, 100 IU vitamin E, 200 mg -carotene, 200 mg Zn methionine, and 1.5 mg Se. Experimental group 2 was administered a vitamin complex (100 g/day) consisting of 100 IU vitamin E and 200 mg -carotene. In the case of multiparous KNC, the estrus detection rate in the control group was 90.0% and those in the experimental groups were 75.0% to 95.0%. However, the first-service pregnancy rate after parturition in the control group was 41.2%, which was significantly lower than that in the experimental groups (71.0% to 76.7%; p<0.05). The average duration of pregnancy in the group supplemented with the vitamin complex was days, which was similar to that in the case of the control group. The birth weight of calves from cows fed with vitamin complex was 25.3 to 27.0 kg, which was similar to that in the case of the control group (25.2 to 26.0 kg). In the case of primiparous KNC, no differences in the estrus rate, pregnancy rate, gestation length, or the birth weight of calves were noted between the groups. Thus, dietary supplementation vitamin and mineral complex have no effect on the reproductive efficiency in primiparous cows, but the pregnancy rate was observed to have increased in multiparous KNC with these supplements.

본 연구는 체세포 복제 한우 송아지의 생산에서 생시체중과 생존성과의 관계에 대하여 살펴보고자 실시하였다. 293두의 한우 대리모에 580개의 복제란을 이식하였다. 복제란의 임신율은 이식 후 50일까지 72.3%로 높았으나, 이후 급격하게 감소하였다. 평균 임신 기간은 복제 송아지에서 287일(279~295일)이었으며, 인공수정 송아지도 287일(255~293일)로 각각 나타났다. 복제 송아지의 생시체중(30.3kg)은 인공수정 송아지(23.7kg)에 비하여 유의하게 높은 것으로 나타났다(p<0.05). 자연분만(n=17, 29.9kg)과 제왕절개(n=14, 32.3kg)로 태어난 복제 송아지의 생시체중은 차이가 없었다. 하지만, 생후 175일 이전에 사망한 복제 송아지(n=18, 32.8kg)의 생시체중이 175일 이상 생존한 복제 송아지(n=11, 28.3kg)보다 유의적으로 높게 나타났다(p<0.05). 흥미로운 점은 생시체중이 15kg 이하(n=5) 또는 35kg 이상(n=9)인 복제 송아지들은 모두 생후 175일 이전에 폐사하였다. 생후 175일 이전에 폐사한 복제 송아지들(n=20)의 사망 원인은 미성숙 개체 2두(10.0%), 폐와 간 이상 2두(10.0%), 폐의 원인 4두(20.0%), 기형 4두(20.0%), 출생 후 급사(sudden death syndrome) 4두(20.0%) 및 기타 원인불명이 4두(20.0%) 등으로 분류되었다. 이상의 결과를 종합하여보면, 복제 송아지의 정상 생시체중이 6개월 이상을 생존하는데 가장 중요한 요소들 중의 한 가지임을 확인하였다.

Cloned calves derived from somatic cell nuclear transfer (SCNT) have been frequently lost by sudden death at 1 to 3 month following healthy birth. To address whether placental anomalies are responsible for the sudden death of cloned calves, we compared protein patterns of 2 placentae derived from SCNT of Korean Native calves died suddenly at two months after birth and those of 2 normal placentae obtained from AI fetuses. Placental proteins were separated using 2-Dimensional gel electrophoresis. Approximately 800 spots were detected in placental 2-D gel stained with coomassie-blue. Then, image analysis of Malanie III (Swiss Institute for Bioinformatics) was performed to detect variations in protein spots between normal and SCNT placentae. In the comparison of normal and SCNT samples, 8 spots were identified to be up-regulated proteins and 24 spots to be down-regulated proteins in SCNT placentae, among which proteins were high mobility group protein HMG1, apolipoprotein A-1 precursor, bactenecin 1, tropomyosin beta chain, H+-transporting ATPase, carbonic anhydrase II, peroxiredoxin 2, tyrosine-rich acidic matrix protein, serum albumin precursor and cathepsin D. These results suggested that the sudden death of cloned calves might be related to abnormal protein expression in placenta.

The development potential of bovine somatic cells was evaluated using nuclear transfer. A single donor cell derived from fetus of HanWoo(Korean Native Cattle) was selected and deposited into perivitelline space of each enucleated oocyte before electrical fusion and activation. Nuclei of donor cells starved for 7 days (37%) tended to support the development of reconstitute embryo the blastocyst stage better than those of donor cells starved 3, 14 and 30 days. The cleavage rate was significantly lower(P<0.05) in reconstitute embryos derived from large size donor cells(51.2%), than those from small medium size donor cells(76.6 and 73.5, respectively). The developmental rate to blastocyst of reconstructed embryos from medium size donor cells was higher than those from small and medium size donor cells. This study demonstrates that an appropriate culture period for induction into quiescent stage and the size of donor cells effect on the efficiency of nuclear transfer using cultured bovine cells.

The objective of this study was to evaluate the embryonic development ability and the appearance of blastocysts of bovine in vitro fertilized oocytes cultured in different culture media, and also to evaluate survival rate after thawing of frozen embryos by using 1.5 or 1.8M ethylene glycol(EG) with sucrose or trehalose. Fertilized oocytes were divided into three groups; i ) monolayer of cumulus /granulosa cell prepared by TGM 199+5% calf serum(TGM199), ii)GRlaa+5% CS, iii)SOF+5% CS, and they were cultured after insemination for 9 days, at 39˚C, under 5% in air, but SOF+5% CS was cultured at 39˚C, under 5% 02, 5% GO2, 99% N2. Blastocysts derived from GRlaa + 5% CS on day 7~8 after insemination were frozen by using 1.5M EG or 1.8M EG with/without 0.2M sucrose or O.1M trehalose. The development rate of blastocysts on day 7 after insemination in SOF+5% CS was significant higher than in TCM199 or CR1aa(P<0.05). The appearance rate of blastocysts on day 7-8 after insemination was higher than in TCM199, when fertilized oocytes were cultured in GRlas or SOF. The survival rate of frozen blastocysts after thawing tended to increase, when blastocysts were frozen by using 1.8M EG with 0.2M sucrose or O.1M trehalose. These results indicated that SOF or CRlaa media with amino acids was superior to TCM199 with monolayer in terms of blastocyst development in culturing of in vitro fertilized bovine nocytes, and sucrose or trehalose was supposed to prevent embryos from the freezing shock.

This study was carried out to enhance the efficiency of Korean Native cattle embryos and establish the techniques for producing the twin calves. Bisected embryos without zona pellucida which were divided by simple method not using holding pipette or whole two embryos were transferred to recipients.The pedigrees of monozygotic twin calves produced by transfer of bisected pair embryos were identified. The results obtained were as follows ; The average successful bisection rate was 89.16%. The embryos of blastocyst stage (91.66%) were bisected successfully at significantly (P<0.05) higher rate, compared with the morula stage embryos (86.66%). The average survival rate of bisected embryos following 24 hours culture was 59.02%. The survival rate of morula stage embryos (62.50%) was significantly (P<0.05) higher than that of blastocyst stage embryos (55.5%). For the production of monozygotic twin calves, ten pairs of flesh or frozen demi-em- lymphocytes antigen, the twin calves produced by transfer of bisected pair embryos of Korean Native cattle were identified in pedigrees and confirmed as monozygotes.

This study was carried out to establish the techniques for producing the calves of genetically superior Korean Native cattle by transfer of frozen-thawed embryos. The effects of some factors related to embryo recovery following superovulation and pregnancy rate following transfer of frozen-thawed embryos were evaluated. Also calving state was investigated. The results obtained were as follows ; The mean number of total and transferrable embryos recovered per superovulated cow was 8.72 and 4.90, respectively, from a total of 72 superovulations using 34 donor cows. There were no significant differences in the number of total or transferrable embryos recovered per superovulated cow between products of follicle stimulating hormone (FSH), years, seasons, and collection numbers. The pregnancy rate was found 44.44% following transfer of frozen-thawed embryos of Korean Native cattle to a total of 180 recipient cows including 82 Angus, 27 Charolais, 62 Hereford and 9 Korean Native cows. The pregnancy rate was significantly (P<0.05) higher in the transfer of excellent (42.99) and good embryos (40.17%), compared with fair (5.90%) grade embryos. And the pregnancy rate was significantly (P<0.05) higher in the transfer of embryos of morula stage (43.86%) than blastocyst stage (15.51%). But there were no significant differences in pregnancy rates between natural and induced estrus estrus asynchrony of 1 days, breeds, and parities of recipient cows. The normal calving rate of 80 pregnant cows following transfer of frozen4hawed em-bryos was 87.5% and the other 10 pregnant cows showed abortion during the period from pregnancy diagnosis at 50~60 days to calving. The average gestation length of normally delivered recipients was 288.50 days and the average birth weight of 70 calves born was 24.22 kg. The gestation length was significantly (P<0.05) shorter in the recipients delivering female calves (286.70 days) than males (289.39 days). But there were no significant differences in gestation tength and birth weight of calves born between the recipient breeds.