This study was conducted to investigate the survival rate of frozen-thawed spermatozoa in equine by glycerol concentration and freezing speed. Two stallions (1 Thoroughbred-13 year old and 1 Arab-7 year old) bred in Korea Racing authority was examined for 1 times in a couple of weeks. Semen was collected by condom method standing heated mare and were centrifuged 650 g for 15 min. and isolated the seminal plasma. Thick fraction of semen was diluted EDTA-Lactose-egg yolk diluents to 1:1 and contained in 0.5 ml straw as 6~14×107cells/ml. Final concentrations of glycerol were 3, 5 and 7% in cryopreseved diluents and added 4 times for 2 hours equilibration. For the freezing, equilibrated straws were located 3 or 5 cm above LN2 gas for 5 or 10 min. Survival rates of pre-frozen sperm were 65.0±13.2%, 68.3±10.4%, 66.7±11.5% and post-frozen were 53.3±23.1%, 45.0±15.0%, 50.0±18.0% in 3, 5, 7% glycerol concentration, respectively. There was no difference between glycerol concentrations. Survival rates of frozen-thawed sperm on freezing speed were 36.7±10.4%, 40.0±7.1%, 30.0±13.2% at 3 cm-5 min and 33.3±11.5%, 31.7± 2.9%, 21.7±10.4% at 3 cm-10 min in 3, 5, 7% glycerol concentration, respectively. Survival rates of frozen-thawed sperm on freezing speed were 43.3±15.3%, 32.0±17.9%, 22.3±15.7% at 5cm-5 min and were 47.5±15.0%, 43.3±12.6%, 48.3±15.3% at 5cm-10 min in 3, 5, 7% glycerol concentration, respectively. There were significantly different between groups (p<0.05). These results suggest that glycerol concentration did not affect cryopreservation of stallion semen within 3~7% but freezing speed affects. In our experiment, the best cryopreservation condition was at 5 cm above LN2 gas for 10 min for pre-freezing and 7% of glycerol concentration. These results lead to commercial AI with frozen-thawed stallion semen.
해충에 대한 화학농약의 오남용으로 인하여 해충의 저항성이 발달되고 환경오염, 인축에 대한 위험 등의 이유로 화학농약의 사용을 줄이게 되었다. 그래서 친환경적인 방제인자인 미생물농약 Bacillus thuringiensis(이하 B.t)가 현재 전 세계적으로 널리 사용되고 있다. B.t는 그람 양성의 호기성 간균으로 토양, 곡물의 분진, 낙엽, 곤충의 사체등 다양한 곳에서 분리되는 것으로 알려져 있는데 본 연구에서는 풍뎅이류 곤충의 사체로부터 B.t를 분리하여 딱정벌레목 및 나비목 해충에 대한 생물활성 검정을 하여 살충활성이 우수한 균주를 선발하였으며 SDS-PAGE를 수행하였다. 파밤나방에 대한 생물활성 검정 결과, CAB530 균주는 처리 48시간 후에 90%가 넘는 사충률을 보여주었으나 CAB533 균주는 6%의 사충률을 보여주었다. 반면에 장수풍뎅이 유충과 배추벼룩잎벌레 성충에 대해서는 살충활성을 나타내지 않았다. SDS-PAGE 수행 결과 파밤나방에 살충활성이 있는 CAB530 균주는 B.t subsp. kurstaki와 비슷한 130kDa의 밴드를 나타냈으며 파밤나방 중장액으로 소화를 시킨 후에 약 65kDa의 밴드를 확인할 수 있었다. 파밤나방에 대해 살충활성이 적은 CAB533 균주는 65kDa의 독소단백질 밴드를 나타냈지만 파밤나방에 대해서 살충활성이 높지는 않았다.