凡纳滨对虾人工授精技术初步研究
Preliminary study on artificial insemination techniques for Litopenaeusvannamei
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摘要: 【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2×107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。Abstract: ObjectiveEffective artificial insemination methods for Litopenaeus vannamei were screened out to provide references for the subsequent gynogenetic induction research. MethodWith 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.ResultThe 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 2×107/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. ConclusionSemen 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.