참게(E. sinensis)의 유생 생장에 미치는 기초적인 환경 및 생물학적 정보를 얻기 위해 본 실험을 실시하였다. 참게 유생을 카드뮴과 수은의 0.1, 0.2, 0.3 ppm농도에 처리하였다. 참게의 생존율과 카드뮴과 수은의 중금속 농도와 양의 상관을 나타내었다. 그런데 96시간 사육시 카드뮴과 수은의 반수치사농도는 유생 단계간 유의한 차이를 나타내었다. 반수치사농도는 카드뮴이 수은보다 높았으므로 수은이 더 치명적이였다. 중금속의 축적은 카드뮴과 수

The aims of these study were to diagnose early pregnancy and reproductive disorders by using progesterone concentration and ultrasonography. The measurement of blood progesterone (P) concentration was conducted to diagnose pregnancy and to detect corpus luteum (CL) or evaluate disorder of CLs. As a result, the incidence rates of reproductive disorders were as follows : SH and EED (41.9%), inacitve ovaries (32.6%), follicullar cyst (9.3%), PCL (7.0%), endometritis (4.7%), pyometra (2.3%) and luteal cyst (2.3%). 61 Cows having Pconcentration 1.0 ng/ml(at the insemination) were increased to 1.0 ng/ml 6day after insemination. 50 cows among 61 cows were diagnosed pregnant. 8 cows among 13 HanWoos having Pconcentration 1.0 ng/ml at the insemination and 1.0 ng/mnl 6 day after insemination had non-ovulatory estrus and the others had Pconcentration 1.0 ng/ml at the insemination and 1.0 ng/ml 6 day after insemination, which was the error of estrus detection. All 13 cows were diagnosed non-pregnant. 47 cows diagnosed pregnant after insemination of Pconcentration 3.0 ng/ml were examined by ultrasonography at 30 day post-insemination. As a result, 41 cows were diagnosed pregnant (87.2%) but 14 cows having Pconcentration 3.0 ng/ml at 21 day after insemination was diagnosed to non-pregnancy. Calving intervals by surveying 100 cows were as follows 11~12 months (20%), 12~13 months (36%), 13~14 months (19%), 14 months (25%), respectively. In conclusion, hormone and ultrasonography help to detect reproductive disorders exactly and diagnose early pregnancy. This study suggest that diagnosis of early pregnancy and reproductive disorder by blood Pconcentration and ultrasonography improve reproduction management of HanWoo.

The aims of this study are to establish a stable isolation method of blastomeres from bovine early embryos and examine their developmental potential in vitro Early embryos were produced by maturation and fertilizaion in vitro of bovine follicular oocytes. Blastomeres were isolated from 2~8-cell embryos in +-, +-free PBS+EDTA after removing the zonae pellucidae Isolated blastomeres were cultured in CZB containing BOEC for upto 240 hpi. Cleavage rates of them were 18.5%(10 /54) in 1 /2 blastomeres, 33.3%(16/48) in 1/4 blastomeres and 34.2%(14 /41) in 1/8 blastomeres, respectively. The rates of blastocystic vesicle formed were 8.7%(4 /46) in 1/2 blastomeres, 26.6% (17/64) in 1/4 blastomeres and 10.3%(8 /78) in 1/8 blastomeres, respectively. Blastomeres developed into various types of blastocystic vesicles and trophoblastic vesicles as evidenced by the Hoechst 33258 staining and morphology. This results suggest that the isolation method used and subsequent culture of isolated blastomeres from bovine early embryos should be useful to obtain extra embryonic cells for various analyses such as PCR and putative ES cell culture.

This study was conducted to compare the insemination time of bovine oocytes and determine the effects of glucose(1.5 mM) on the development of bovine embryos at early cleavage stage. Oocytes were matured for 24 h, followed by exposure to sperm and cultured in modified Tyrode's media drops or with bovine oviduct epithelial cell monolayer prepared in TCM199(BOECM). Insemination time and culture system were varied in each experiment. In experiment 1, to investigate the developmental capacity of bovine embryos after different time of exposure to sperm, bovine ova and sperm were co-incubated for 18, 30 or 54 h, respectively. The development to blastocysts of 30 and 54 h insemination groups were significantly higher(P<0.05) than 18 h group, and in case of blastocysts of cleaved embryos, 30 h group were significantly higher(P<0.05) than other groups. In experiment 2, we investigated the effect of glucose on early bovine embryos. After 18 h insemination, in vitro fertilized oocytes were separated following 3 groups ; G+0, C+24 and C+48. Oocytes of G+0 group were cultured in glucose added Tyrode's medium after fertilization, oocytes in C+24 and C+48 groups were cultured in glucose free Tyrode's medium after fertilization. After 24 h culture, G+24 group was moved to glucose added medium. All oocytes of 3 groups were moved to BOECM after 48 h culture. The rates of cleavage and development to blastocysts in G+0 group were significantly lower than other groups. In experiment 3, we determined the effects of glucose exposure from 8 to 20 h after insemination on the cleavage and development of oocytes. The oocytes in glucose added group had high capacity of cleavage and further development. This study shows that in bovine oocytes, the optimal exposure to sperm is 30 h and glucose exposure to bovine one-cell embryos is detrimental to their first cleavage and further development in vitro but there has no evidence of detrimental effect of glucose(1.5 mM) exposure to bovine embryos over the two-cell stage in vitro.

The present study was conducted to investigate the influence of embryo cell stage at 18h post-fusion and oocyte preactivation on sebsequent in vitro developmental potential in the nuclear transplant rabbit embryos. The embryos of 16-cell stage were collected and synchronized to G phase of 32-cell stage. The recipient cytoplasms were obtained by removing the first polar body and chromosome rnass from the oocytes collected by non-dis-ruptive microsurgery procedure. The separated G phase blastomeres of 32-cell stage were injected into non-preactivated recipient cytoplasms. Otherwise, the enucleated recipient cytoplasms were preactivated by electrical stimulation at 18h post-hCG injection and the separated G phase blastomeres of 32-cell stage were injected. Mter culture until 20h post-hOG injection, the nuclear transplant oocytes were electrofused by electrical stimulation. The fused nuclear transplant embryos were classified into 3~4-cell, 2-cell and 1-cell stage at 18 hrs post-fusion and cultured until the embryos reached blastocyst stage. The developmental rate to blastocyst stage was significantly (P <0.05) higher in all the reconstituted embryos of 3~4-cell stage(58.0%) than in 2 and icell stage. The developmental rate to blastocyst stage in the embryos of 3~4-cell stage at 18 hrs post-fusion was significantly (P<0.05) higher in the reconstituted without oocyte preactivation(77.8%) than in the oocyte-preactivated embryos (33.3%). These results indicated that the higher rate of in the in vitro development to blastocyst stage might be obtained form the embryos which were reconstituted with nuclear donor of G phase and non-preactivated oocyte, and developed more rapidly for 18 hrs post-fusion.