국내에 서식중인 청개구리의 배아를 활용하여 농약류 3종의 독성평가를 위해 FETAX(Frog Embryo Teratogenesis Assay-Xenopus) 기법에 따라 청개구리(Hyla japonica)의 배아를 배양하면서 Benomyl(살균제), Carbofuran(살충제), Thiobencarb(제초제)의 효과를 probit 분석법으로 조사하였다. 그 결과, Benomyl, Carbofuran, Thiobencarb의 농도에 의존하여 유생의 체장 길이는 감소하고 치사율과 기형율은 증가하였다. Benomyl, Carbofuran, Thiobencarb의 teratogenic concentration(EC50)은 각각 1.00, 0.58, 4.75mg/L을 나타내어 Carbofuran이 기형유발에 가장 민감하게 반응하였으며, embryo lethal concentrations(LC50)은 7.04, 28.71, 16.12mg/L을 나타내어 Benomyl이 가장 낮은 농도에 서 배아를 치사하는 것으로 나타났다. Teratogenic index(TI=LC50/EC50)는 Benomyl 7.04, Carbofuran 49.50, Thiobencarb 3.39를 나타내어 TI값이 모두 기형유발물질로 판단하는 기준인 1.5 이상을 나타내어 시험에 사용된 농약류 3종은 최기형성 물질로 판단되며 Carbofuran이 가장 강력한 최기형성물질로 작용함을 알 수 있었다.

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 On the 3 day administration of FSH, 25 mg was administered and CIDR was withdrawn.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.