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引用本文:蔡明夷,柯才焕,王桂忠,郭 峰,郑 云,游伟伟,许贻斌.九孔鲍精子短期保存技术研究[J].海洋科学,2008,32(1):1-5.
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九孔鲍精子短期保存技术研究
蔡明夷1,2, 柯才焕1, 王桂忠1, 郭 峰1, 郑 云1, 游伟伟1, 许贻斌1
1.厦门大学海洋系;2.集美大学水产生物技术研究所
摘要:
应用单因素试验和三因素正交试验研究了温度、二甲基亚砜(DMSO)浓度及精子密度等因素对九孔鲍(Haliotis diversicolor supertextexta)精子短期保存的影响。单因素试验结果表明: DMSO、维生素C、甘油、葡萄糖等添加剂中仅5%和7. 5% DMSO有利于精子活力的保持; 3个试验温度中28℃精子活力下降速度最快,其次为1℃, 最慢是5℃; 稀释度越高精子活力下降速度越快。正交试验中, 九孔鲍精液分别在25个保存条件下保存96 h, 保存期间每隔一段时间取样观察精子活力。25 个处理的活力变化数据用生长函数拟合, 根据拟合方程计算出精子活力半衰期, 并对结果进行直观分析及方差分析。结果表明, 较理想的精子保存条件是: 温度为7℃, DMSO质量分数为3%, 精子密度为7.5×108个/mL; 各因素对精子活力半衰期的影响按精子密度、温度、DMSO密度顺序递减; 精子密度对精子活力半衰期有显著影响(P<0.05) , 温度及DMSO浓度的影响不显著(P>0.05) 。
关键词:  九孔鲍(Haliotis diversicolor supertextexta)  精子保存  正交试验  精子密度  二甲基亚砜  温度
DOI:
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基金项目:国家863 计划资助项目(2003AA603240)
Semen short-term storage of the small abalone
郭 峰,郑 云
Abstract:
The effects of sperm concentration, temperature and DMSO concentration on the decay of sperm viability of the small abalone, Haliotis diversicolor supertextexta were determined. The results of one-factor experiment showed: All the test extenders had negative effects on the life of small abalone sperm,except for 5% DMSO and 7.5% DMSO; Among the three test temperatures, the rank of decay speed of sperm motility score was 28,1,5℃; The decay speed increased with the dilution increase.A 3-factor and 5-level orthogonal experiment was designed based on the results of one-factor experiment. 25 aliquots of small abalone semen were stored separately for intervals up to 96 h. The data of sperm viability score in 25 given conditions were fit into 25 equations with a growth model. The half-lives of sperm in the 25 kinds of storage conditions were estimated with the equations. The results show that the optimal store condition was at sperm concentration of 7.5×108 sperm/ mL in seawater with 3 % DMSO at the temperature of 7℃. Among the three factors, the most important one was sperm concentration,and then storage temperature, the last one was DMSO concentration. The results of ANOVA showed that the effect of sperm concentration on half life of sperm was significant(P<0.05), while the effects of temperature and DMSO concentration were not significant.
Key words:  Haliotis diversicolor supertextexta  sperm storage  orthogonall experiment  sperm concentration  DMSO  temperature
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