凡纳滨对虾人工授精技术初步研究

Preliminary study on artificial insemination techniques for Litopenaeus vannamei

【目的】筛选出有效的凡纳滨对虾人工授精方法，为后续的雌核发育诱导研究奠定基础。【方法】以受精率和幼体孵化率为考核指标，探讨外体精卵结合授精法、精荚移植法、精液移植法等不同人工授精技术对凡纳滨对虾繁育效果的影响。【结果】体外精卵结合授精法在各处理组均无幼体成功孵出，但在精子密度为2＆#215;107个/L和卵子密度为300枚/L，且在卵排出1 min内完成授精操作时，可获得相对较高的受精率（6.67%）。在精液移植法中，发现精荚在8℃恒温条件下保存3h内进行授精操作，仍可获得一定的受精率和幼体孵化率，超过3h后则无法实现受精；精荚保存温度以8-16℃为宜，保存温度升至25℃时则无法实现受精。3种人工授精技术中，以精液移植法的受精率和幼体孵化率最高，分别为52.33%和12.35%，其次是精荚移植法，体外精卵结合授精法的效果最差，无幼体孵出。【结论】精液移植法是凡纳滨对虾最有效的人工授精方法，但精荚保存时间不宜超过3 h，且应在雌虾排卵后1-2 min完成挤精荚和网搓释放精子溶液，同时可添加预先收集浓缩制备好的卵水溶液，以提高凡纳滨对虾的人工授精效率。

[Objective]Effective artificial insemination methods for Litopenaeus vannamei were screened out to provide references for the subsequent gynogenetic induction research. [Method]With fertility rate and hatching rate as evaluation in-dexes, the effects of fertilization in vitro spermatophore transplantation and semen transplantation on the breeding efficiency of Litopenaeus vannamei were compared.[Result]The results indicated that no larva was hatched from all treatments in this ex-periment, but relatively high fertility rate （6.67%） could be gained under sperm density of 2＆#215;107/L and unfertilized egg density of 300 individual/L within unfertilized egg effluxtime time of 1 min. In semen transplantation method, certain fertilization rate and hatching rate could be obtained with spermatophore at 8℃preservation and insemination operating within 3 h. Fertiliza-tion failed to achieve under more than 3 h. The appropriate spermatophore storage temperature was 8-16℃. Fertilization could not be achieved above 25℃. Among the three kinds of artificial insemination methods, semen transplantation method had the highest fertilization rate and hatching rate, which was 52.33% and 12.35%, respectively. It was followed by spermatophore transplantation method, and vitro fertilization method was the lowest with no nauplius hatched out. [Conclusion]Semen trans-plantation was the most effective artificial insemination method for Litopenaeus vannamei, and the spermatophore storage time should not exceed 3 h. Spermatophore extrusion and sperm solution release should be performed within 1-2 min after ovulation. Meanwhile the prepared egg collection solution could be added to improve the artificial insemination rate.