This study was done to examine the relationship between serum chemical values (urea nitrogen, glucose, total protein and cholesterol) of recipients and pregnancy rate following embryo transfer. Blood samples were taken from 184 Holstein heifers or cows on Day 6 or 7 (Day 0=day of estrus) to analysis for serum urea nitrogen, glucose, total protein and cholesterol concentrations. After selection of recipients, frozen Holstein embryos were thawed and directly transferred to recipients non-surgically. The average serum concentrations of urea nitrogen, glucose, total protein and cholesterol were 13.8 mg/dl, 56.5 mg/dl, 7.2 mg/dl, 124.8 mg/dl, respectively. The average concentrations of serum urea nitrogen and cholesterol were lower (P<0.05) in pregnant recipients (10.7 mg/dl, 99.2 mg/dl) than in non-pregnant recipients (13.0 mg/dl, 122.2 mg/dl), respectively, although the concentrations of glucose and total protein were not different. These results show serum urea nitrogen and cholesterol concentrations can be used important factors for selection of recipients in Holstein.

To investigate the effect of co-transfer of trophoblastic vesicle (TV) with frozen-thawed in vitro Produced (IVP) bovine embryo on pregnancy rate, IVP blastocysts were transferred to synchronized recipients. Elongated blastocysts were recovered at Day 13 to 15, and dissected more than 4 pieces to removed the embryonic disc. Throphoblastic fragments were cultured for 48 hours to make throphoblastic vesicles (TVs). TVs were cryopreserved in ethylene glycol or vitrification solution and frozen-thawed TVs were co-transferred to recipients with frozen-thawed IVP embryos. 1 The recovery rate of elongated blastocyst on Day 13 to 15 was 22.5% (18/80) and the size of recovered elongated blastocysts was 0.2∼5.0mm. 2. Eighteen elongated blastocysts were dissected into 88 pieces and 61.4% of those pieces were formed to TV (54/88) 3. The viability of frozen-thawed TV in ethylene glycol was higher than in vitrified solution (92.8% vs. 68.8%) 4. The pregnancy rate in co-transfer with frozen-thawed TV and IVP blastocyst was better than transfer only IVP blastocysts (50.0% vs. 23.1%).

The ovaries of 178 Holstein heifers or cows (heifer; 41, 1 parity; 72, 2 parity; 65) on Day 6 or 7 (Day 0=day of estrus) were examined by transrectal ultrasonography. Diameter of corpus luteum (CL) and large follicle ( 10 mm), and luteal tissue area were determined by ultrasound system with a 5 MHB rectal probe. Blood samples were taken to progesterone analysis. After selection of recipients, frozen Holstein embryos were thawed and directly transferred to recipients non-surgically. The diameter of CL and luteal tissue area was greater (P<0.01) on Day 7 than on Day 6 in heifers, 1 parity or 2 parity cows, respectively, although progesterone concentrations were not different. The presence of fluid-filled luteal cavitied or multiple CL (2 or more) did not affect serum progesterone concentration. A large follicles were observed in 67.4% of heifers or cows and the average diameter was 14.1 mm. Greater luteal tissue area attributed higher pregnancy in heifers, but not in cows, although there were no difference on pregnancy rate according to progesterone concentration in heifers or cows. The pregnancy rate of recipients contained a large follicle at embryo transfer was lower than that of recipients not contained. These results show ultrasonic assessment of ovaries in Holstein recipients is a reliable tool to determine the follicle and CL for recipient selection.

This study was undertaken with three objectives : to determine the optimal time of blood collection for plasma urea nitrogen(PUN) analysis, to examine the frequency distribution of PUN levels in recipient herd and to relate concentration of PUN to conception rate in Hanwoo recipients. The relationship between PUN level and time postfeeding was examined for 5 individual cows. Mean concentration of PUN rose for 4hrs postfeeding and decreased to PUN level before feeding at 10hrs postfeeding. And then, the blood for PUN analysis was collected at the time before feeding in next experiments. The ratio of cows with PUN concentration of < 12, 12∼18 and 18mg/dl were 50.6, 39.9 and 9.5% in 163 recipients, individually. The pregnancy rate of cows with PUN concentration 12∼16 mg/dl (63.3%) was higher than that of cows with PUN concnetration < 12 mg/dl (46.7%) or > 16mg/dl (42.9%). There results suggest that the PUN test may be beneficial for management of recipient herd in effects to maintain or improve reproductive efficiency.

This study was carried out to establish an effective system for embryo transfer techniques by analyzing several factors affecting in-vivo embryo transfer in Korean cattle Embryos were transferred into a toral of 301 recipients. The results obtained in studies on the factors affecting pregnancy rate after embryo transfer by condition of transfer time were as follows ; 1. The pregnancy rate by the seasons of transferred fresh and frozen embryos were not different, but the pregnancy rate was slightly higher in summer(80.8%). 2. The pregnancy rate by the days of embryo transfer after estrus were not different when fresh embryos were transferred, but the pregnancy rate was highest at 8 days when frozen embryos were transferred(P<0.01, 40.0%). 3. The pregnancy rate at estrus synchronization was remarkably higher with PGF treated than natural (P<0.05, 70.4%, 43.4%). 4. The pragnancy rate by the degree of estrus synchronization was best when the estrus was synchronized in both fresh and frozen embryos (83.3% and 29.7%, respectively), but the pregnancy rate was not different among 2 days. But the pregnancy rate of frozen embryos were slightly higher when the recipients exhibited estrus earlier than donors.

This study was carried out to establish an effective system for embryo transfer techniques by analyzing several factors affecting in-vivo embryo transfer in Korean cattle. Embryos produced in-vivo were transferred into a total of 301 recipients The results obtained in studies on the factors affacting pregnancy rate after embryo transfer by condition of recipients were as follows ; 1. The pregnancy rate by age and parity of recipients showed high in 5~8 and over 12 years old(72.7~73.9%), and 3rd~4th parity(82.1%) for fresh embryos(P<0.05). The pregnancy rate did not differ by age and parity of recipients in frozen embryos. The pregnancy rate of frozen embryos tended to be similar to that of fresh embryos(38.5% and 25.0~36.7%). 2. The number of observation for normal estrus cycles of recipients did not differ In pregnancy rate between one and 2 times in fresh embryos(64.9%, 69.8%). The pregnancy rate by transferred frozen embryos showed significantly higher after 2 times of observation(P<0.05, 16.3%, 37.5%). The pregnancy rate by days open did not differ between fresh and frozen embryos. But the pregnancy rate was slightly higher in 12 months and 6 months of days open for fresh and frozen embryos, respectively(70.1~71.1% and 24.5%, respectively). 3. The pregnancy rate of transferred fresh and frozen embryos into right and left side of uterine horn did not differ(62.1% : 65.9% 25.0% : 24.3%, respectively). The pregnancy rate by the grade of CL was not different in fresh embryos, but the pregnancy rate was significantly higher in the grade A than B for frozen embryos(P<0.01, 43.2%, 16.2%).

This study was carried out to establish an effective system for embryo transfer techniques by analyzing several factors affecting in-vivo embryo transfer in Korean cattle. Embryos produced in-vivo were transferred into a total of 301 recipients. The results obtained in studies on the factors affecting pregnancy rate after embryo transfer by condition of embryos were as follow ; 1. The pregnancy rate of 301 recipients was 45.2% and higher with fresh embryos than with frozen embryos(63.5% : 21.4%, P<0.01). Embryos superovurated by FSH-P had slightly greater than by SUPER-OV in pragnancy rate, athough these were no difference between two treatments. 2. The pregnancy rates of transferred morulae and blastocysts showed no difference between fresh and frozen embryos(63.5% : 63~6% ; 20.0% : 25.8%). However, the pregnancy rates by quality of flesh and frozen embryos were significantly different(P<0~05). The pregnancy rates were outstandingly high in the grade A, B of fresh embryos(59.0~66.4%), and in the grade A of frozen embryos(43.6%). 3. The number of transferred embryos showed no difference in pregnancy rate, but when frozen embryos transferred, the pregnancy rate was slightly higher with two embryos than that with one embryo.

치밀 난구세포로 둘러싸인 소 난자를 . 5% 배양기에시 5% superovulated cow serum(SCS)이 첨가된 m-TCM 199 medium 으로 시간 배양하였으며, 수정능이 획득된 정자와 체외수정하였다. 7일8일경의 수정란을 1.3M methyl cellosolve(MC), 1.1M diethylene glycol(DEG), 1.8M ethylene glycol(EG), 1.6M propylene glycol(PG) 및 1.1M 1,3-butylene glycol(BG) 용액에서 10분간 평형시킨 후 0.25 ml 스트로내에 장전하였다. 스트로를 의 alcohol bath freezer에 넣고 까지 /분 속도로 냉각, 식빙 후 10분간 정체시켰으며, /분 또는 /분으로 까지 냉각 후 스트로를 액체질소에 침지하여 보관하였다. 수정란이 들어있는 스트로를 온수에서 융해하였으며, 수정란을 TCM 199 medium 으로 옮긴 후 5% SCS가 첨가된 TCM 199 medium 에서 48시간 배양하였다. 수정란이 양호한 형태를 유지하며 나중의 발육단계로 진행된 것을 생존한 것으로 간주하였다. 각 종류의 동해방지제에서 동결된 수정란의 일부는 융해 후 동해방지제를 제거하지 않고 직접 비외과적으로 이식하였다. 동결-융해 후 동해방지제의 종류에 따른 탈출배반포 발달율은 EG 50.0%, MC 53.6%, DEG 56.9%, PG 58.0% 그리고 BG 11.5%였다. /분 또는 /분 으로 냉각한 수정란의 생존율은 두 그룹간에 유의적인 차이가 없었으나 (P<0.05), 탈출배반포 발달율은 -0.5분 /분(22.6%, 12/53)보다 /분(64.6%, 31/48) 냉각시에 유의적으로 높았다(P<0.01). 동해방지제의 종류에 따른 수정란의 수태율은 MC 48%(10/21). DEG 30%(3/10), EG 74%(20/27) 및 PG 40%(4/10) 였다. 이러한 결과로 보아 MC, DEG, EG 그리고 PG는 소의 체외수정란의 동결을 위한 동해방지제로서 이용될 수 있음을 보여주었다.

The objective of this study was to enhance the pregnancy rate of repeat-breeder Hanwoo with gonadotropin-releasing hormone(Gn-RH) at the time, dose and site of administration.The results obtained were summaried as fallows:1.Ovulation time and pregnancy rate following GnRH administration time was 46.0, 27.4, 42.0 and 43.2hr and 33.3, 57.1, 37.5 and 40.0% at non-treatment, estus, 1st A' and 2nd Al treatment, respectively.2. Ovulation in repeat-breeder was induced 100% within 24hr with GnRH administration at the time of estrus.3. Ovulation time and pregnancy rate following GnRH adminstration dose and site was 25.2, 32.6, 17.6 and 27.6hr, and 28.6, 42.9, 75.0 and 66.7% at 50g+IU, 50g+IM, 100g+IU and 100g+IM treatments, respectively. It is concluded that GnRH administration for repeat-breeder was enhanced the pregnancy rate when treated with 100g intrauterine at the time of estrus.

This study was carried out to increase the viability of bovine frozen4hawed in vitro produced (IVP) embryos and pregnancy rate by direct transfer method. Cumulus-oocyte complexes were aspirated from excised Hanwoo ovaries and matured in TGM 199 for 20~22 hours at 38.5 in 2% in air. Matured oocytes were fertilized with capacitated sperm for 6 hours and then co-cultured with cumulus cells for 9 days. 63% of the oocytes cultured was deaved and 29% out of them developed into blastocysts. Good or excellent grade of blastocysts on D 7 or 8 were frozen with 1.8M ethylene glycol as a cryoprotectant for direct transfer. Frozen embryos were thawed at 2 water for 10 sec following 4~5 second in air. For the survival assay of frozen4hawed lVP blastocysts, they were cultured in TCM 199 supplemented with 100M -mercaptoethanol and 20% FCS for 72 hours. The percentage of embryos developed to re-expanded or hatched after 72 hours culture was 95. 5 and 77.3%, respectively. When frozen-thawed Ivp embryos were transferred to 43 synchronized recipients by direct transfer method, eighteen recipients (41.8%) was pregnant. The highest pregnant was in naturafly synchronized recipients (71.4%), but induced estrus by using PRID(29.2%) and PGF(20.0%) was showed lower pregnancy rate. The pregnancy rate was higher in day 7 blastocysts(56.0%) than day 8 blastocysts(22.2%). (Key words: in vitro produced, blastocyst, frozen-thawed, direct transfer)