The present study was carried out for the comparative study on the collection of bovine follicular oocytes by ultrasound-guided ovum pick-up(OPU) and slaughterhouse-derived (SHD) ovary aspiration and in vitro production of bovine embryos with the follicular oncytes in Korean native cows. Bovine follicular nocytes were observed with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use and the oocytes were collected with the aspiration equipment attached to the ultrasonograph. Bovine ovaries were collected and transported in phosphate buffered saline from the local slaughterhouse, the follicular oocytes were collected by the aspiration method. The collected follicular oocytes in good quality were matured, fertilized and cultured in the media. The total number of the visible follicles and the recovery rate of follicular oocytes were increased in ultrasonography following follicle stimulating hormone(FSH) treatment in Korean native cows. The mean recovery rate of oocytes was 66.2, 52.8 and 41.7% in the FSH-OPU, non-treatment-OPU and SHD ovaries, respectively. The mean number of recorved oocytes per cow were not significantly(P<0.05) different between the FSH-OPU(14.011.54) and SHD(17.1i6.21) groups, but the numbers in both groups were significantly(P<0.05) higher than the number in the non-treatment-OPU(3.71.57) group. The mean number of usable nocytes in Grade T /11 per ovary was 6.3, 4.8 and 1.3 in the cows of the SHD, FSH-OPU and non-treatment-OPU groups, respectively. The in vitro developmental rate to the blastocyst was not significantly different between the oocytes obtained via OPU(37.1%) and SHD(29.3%). Therefore, the ultrasound-guided OPU technique can be applied to the production of excellent embryos from the high-quality cows, and for the large scale production of in vitro bovine oocytes and embryos, the SHD ovary aspiration method is valuable.

From September 1993 to August 1997, we treated ovarian disorders in 1,782 repeat breeder cows after diagnosis by ultrasound on 35 farms in Kyeong-ki do. The rates of ovarian appearance were 59.8% of CL group, 16.7% of ovarian atrophy or hypofunction, 15.4% of luteal cyst, 4.3% of follicular cyst and 3.7% of follicle group in diagnosis with rectal palpation and ultrasound. The results of treatment for ovarian disorders were 1,316 cows(73.8%) in estrus, 348 cows(19.5%) in non-detected and 118 cows(6.6%) in unidentified. The rates of PGF, GnRH and mineral vitamin complex treatment to estrus were 79.6, 69.2 and 50.3%. Two groups were treated with 5 ml PGF intramuscular injection(I.M.) and 1.5 ml PGF intraovarian injection(I.O.), and the results of 1.5ml PGF I.0. were significantly higher than that of 5ml PGF I.M. in inductiom estrus(p<0.05). The pregnant rates were 29.8% in total repeat breeder cows with ovarian disorders following diagnosis and treatment. In summary, rectal palpation and ultrasonography were proven to be useful tools of diagnosis and treatment in ovarian disorders, and it was also suggested that the response to treatment with PGF I.0. was better than PGF I.M.

This study was carried out to compare the actual size(length and height) of ovaries, follicles and corpora lutea of Korean native cow with those on sonograms. We used 3 different probes(3.5 MHz abdominal probe, 6.5 MHz transvaginal probe and 5.0 MHz transrectal probe) and a calipher for measurements of ovaries, follicles and corpora lutea on sonograms and actual size. Under water immersion, 157 ovaries were scanned with 3 probes and measured in actual size and compared each other. The average height and width of ovaries of Korean native cows were 17.403.99 and 34.236.02mm, respectively. In comparison of height, length of ovaries and preovulation follicles, we found that image with a transvaginal probe was nearly the same as the actual size(p<0.01), but with an abdominal probe the image was appeared larger than the actual size. In measurement(diameter) of preovulation follicles the transvaginal probe was proven to be more accurate to the actual size than other probes and in corpus luteum measurement all probes were accurate. In the comparison of number of follicles by different size ranges, there was no statistical difference in the count of follicles over 10 mm in diameter between the transvaginal probe and naked eyes.

The present study was carried out to evaluate the effect of superovulation treatments on ovarian responses, oocyte recovery rates and grades of collected oocytes using an ultrasound-guided transvaginal approach in Korean native cows. Superovulation in cows was induced with two different regimenes: 1) FSH-decreasing dose(n=8): the cows were received twice per day for three days of the total dose of 400 mg of FSH-p, 2) FSH-single dose(n=9): the cows were administrated a single dose of 400 mg of FSH-p in 25% PVP. The Observation of visible follicles and collection of oocytes were performed 12 hours following the last FSH in FSH-decreasing dose group and 48 hours after the FSH-single dose injection. All visible follicles larger than 6 mm were punctured and aspirated with a 6.5 MHz convex-array ultrasound transducer designed for intravaginal use. The mean number of visible follicles(> 6 mm) was significantly(P<0.05) higher in the FSH-decreasing dose treatment (22.811.9) and FSH-single dose treatment (20.612.0) groups than the non-treatment group(7.08). The mean recovery rate of oocytes was not significantly(P<0.05) different between the treatment and control groups, but the mean number of collected oocytes was significantly(P<0.05) higher in the FSH-decreasing dose treatment( 12.611.5) and FSH-single dose treatment (11.813.6) groups than the non-treatment group(3.70.5). In conclusion, the FSH-single dose treatment at superovulation in cows for ultrasound-guided aspiration might increase the number of aspiratable follicles and the recovery rate of follicular oocytes as the FSH-decreasing dose treatment.

The present study was carried out to produce in vitro fertilized embryos with immature follicular oocytes collected by transvaginal aspiration from Holstein cows. A simple aspiration apparatus consists of two stainless steel tubes, an inner tube (needle holder; 1.2cmdiameter, 55cm long) and an outer tube (1.5cm diameter, 4Scm long), and a hand-operated vacuum pump was used. Under epidural anesthesia, the needle guide was passed into the vagina of the cow to a point next to the cervix. An ovary was placed against the wall of the vagina over the end of the aspiration needle by rectal manipulation. As the needlepassed into the ovary, an assistant was asked to apply vacuum(l00mrnHg) and the ovary was manipulated back and forth in all directions over the needle. When all sites of the ovary was aspirated, the needle was withdrawn and the needle guide was moved to the other side of ovary and the procedure was repeated. When the oocyte aspiration procedure was finished, collected fluid was transported to laboratory. Oocytes surrounded with at least 1 layer of cumulus cells were matured, fertilized and cultured in vitro. The results were as follows; Ninety seven oocytes were collected by transvaginal aspiration from seventeen Holstein cows(5.7 /head). The number of oocytes surrounded with at least 1 layer of cumulus cells were 60(61.9%). Following in vitro maturation, fertilization and culture, the cleavage and development rate to morula+blastocyst were 83.3% and 30.0%, respectively. From this study, transferable in vitro fertilized embryos could be produced with imma- ture follicular oocytes collected by transvaginal aspiration from Holstein cows using a simple aspiration apparatus

To improve the efficiency of in vitro production of embryos with follicular oocytes in Korean Native cows, the recovery rates, in vitro maturation, fertilization and development, and the time required for collecting and processing oocytes by aspiration with or without slicing were evaluated comparatively. The ovaries were obtained from a local abattoir and placed in physiological saline at 25~28 and brought to the laboratory within 3 hrs. The oocytes were collected by aspiration of follicles(2~6mm) with or without slicing ovaries after aspiration, and classified into Grade I, Grade II, Denuded, Expanded oocytes by the morphology of cumulus cells attached and the homogeneity of cytoplasmic granules. Also the time required for each step of collecting and processing oocytes were measured. The cumulus cells were removed in some Grade I oocytes to measure their size and nuclear configuration before and after in vitro maturation. The Grade I oocytes were matured in vitro(IVM) for 24 hrs. in TGM-199 supplemented with 35g /ml FSH, 10g /ml LH, 1 g /ml at 39 under 5% C02 in air. They were fertilized in vitro(IVF) by epididymal spermatozoa treated with heparin for 24hrs. and then the zygotes were cocultured in vitro (IVC) with bovine oviductal epithelial cells for 10 days. The results obtained were as follows: The number of oocytes recovered per ovary was averaged 6.6 by aspiration and 11.2 by slicing post aspiration, which summed to 17.8. The number of Grade I oocytes recovered per ovary was averaged 3.1 by aspiration and 3.6 by slicing, which summed to 6.7. The percentage of Grade I to total oocytes recovered was significantly(P<0.05) higher as 48.0 % in aspiration than 31.6% in slicing post aspiration. The time requlred for recovering a Grade I oocyte by aspiration and slicing was 1.1 and 2.5 min, respectively. The mean diameter of Grade I oocytes by aspiration and slicing was similar as 148.7 and 151.5m, respectively. The percentage of Metaphase II stage oocytes after IVM for 24 hours was significantly (P

본 시험은 국내생산 주요 조사료인 볏짚, 옥수수사일레지, 목건초를 급여하면서 농후사료 수준을 조절하였을 때 반추생리 및 저작형태에 미치는 효과를 조사하기 위하여 농후사료 5개수준(0, 3, 6,9 12kg/일)에 젖소 15두를 공시하여 시험을 실시한 결과는 다음과 같다. 1. 건물섭취량은 볏짚, 옥수수사일레지, 목건초가 대사체중비 4.97, 5.47 그래고 5.60%를 섭취하였으며 농후사료 증가에 따른 조사료 건물섭취량 감소량은 볏짚 0.330, 옥수수

The suitable electric stimulation is essential for activation and fusion of oocytes before or after nuclear transplantation The present study was undertaken to determine the optirnal condition for the parthenogenetic activation of in vitro rnatured(IVM) bovine oocytes by electric stimulation. Different direct current(DC) electric voltage of 1.0, 1.5 and 2.0 kV/cm and pulse duration of 30, 60 and 120 sec were applied to the JVM nocytes in 0.3 M mannitol solution containing each 100 M CaCl and MgCl. IVM occytes at 24, 28 and 32 hours Post-maturation(hpm) were also electrically stimulated at 1.5 kV /cm, for 60 sec. The stimulated nocytes were then co-cultured in TCM-199 solution containing 10% fetal calf serum with bovine oviductal epithelial cells for 7~9 days in a 5% incubator at 39 ~ Their activation and in vitro development to morula and blastocyst were assessed under an inverted microscope. The higher activation rates 62.8 and 63.4% and in vitro de- velopment rates to morula and blastocyst 5.1 and 10.9% were shown in the oocytes stimulated at the voltage of 1.0 and 1.5 kV/cm than 2.0 kV/cm, respectively. No signifi- cantly(P<0.05) different activation rate was shown in JVM oocytes stimulated for 30, 60 and 120 sec, but developmental rates to morula and blastocyst was significantly(P<0.05) higher in the oocytes stimulated for 30 sec(6~3%) and 60 sec(10~0%) than 120 sec(0~ 0%). The aged oocytes at 28 and 30 hpm showed significantly(P<0.05) higher activation rates(72~7 and 79.7%) than the oocytes at 24 hpm(50~9%)~ Also, their developmental rates to morula and blastocyst were significantly(P<0.05) higher in the nocytes at 28(14.3%) and 32 hpm(15.9%) than 24 hpm(3.6%). From these results, it can be suggested that the optimal electric stimulation for IVM bovine occytes is a DC voltage between 1.0 and 1.5 kV/cm, pulse duration of 30 or 60 sec, and the optimal age of IVM oocytes for electric activation is at 32 hpm.

대상방목 체계하에서 Lolium perenne초생의 채식 허용량을 3 수준으로 달리 하여 고능력 착유우에 의한 방목시험으로부터 얻은 채식량과 산유량을 조사하여 얻을 결과를 요약하면 아래와 같다. 채식된 사포로부터 배설된 분증질소, 크롬 및 회분 함량은 시험시기가 늦을수록, 채식허용량이 낮을수록 약간 감소되는 경향이었다. 시험시기별 채식 허용량의 수준에 따른 채식초의 소화율은 시험시기 1의 82.7%에서 시험시기 3의 79.1%로 감소하였고, 고수준의 채

방목체계하에서 일당 채식허용량(kg OM/cow)의 수준에 따른 일당 채식량 및 산유량의 경향을 방목에 따른 초생의 구성 및 구조적 변화와 관련하여 검토하는데 목적을 두었다. 본 보에서는 5일간 24시간 대상 방목체계하에서 고능력 착유우에 대한 3 수준의 일당 채식허용량에서 방목후 초생의 구조와 관련된 요인들의 변화를 조사한 결과를 요약하였다. 시험시기별 방목적 초생량 및 초장은 시험시기 1에서 5,047kg OM/ha와 341mm에서 초생의 생육이 진

Total thirty of flushing were attempted on day 4 to 15 of estrus cycle with S heifers and 9 cows by nonsurgical method. The flushed or recovered rate among flushings was 86.7% (26/30) or 88.5% (23/26), respectively. There was no difference in the recovered rate between heifers (85.7%,6/7) and cows (89.5%, 17119). The embryo was recovered on day 4 to 15 of estrus cycle from the donors in natural heat without any technical difficulties.The I2FG Foley catheter used for pubertal heifers had sometimes plug in it with uterine mucus during flushing of uterine horn. But the problem could be overcomed by pumping the catherter with fluthing solution or by changing the catheter. Three normal embryos were recovered from 3 pubertal (10-11 month old) heifers. The rate of normal and abnormal eggs was 60.9% (14123) and 39.1% (9/23), respectively. The abnormal eggs were on degenerating except one unfertilized egg and were mostly recovered from heifers or cows flushed consecutively during the estrus cycle. The developmental states of normal embryos were l6-cells on day 5, 32-cells on day 6, compacted-morula on day 7, early-to expanded-blastocyst on day 8-to 9, and hatching-to hatched-blastocyst on day 10 to 11 of estrus cycle. The stage of embryos on day 8 to 10 showed varities among donors. On day 8 to 9 of estrus cycle hatching-blas tocyst was recovered from some donors.