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New methods for cryopreservation of chicken semen using Hiroshima solution (trehalose solution)

使用广岛溶液（海藻糖溶液）冷冻保存鸡精液的新方法

基本信息

批准号：
02660279
负责人：
TERADA Takato
金额：
$ 1.28万
依托单位：
Hiroshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1990
资助国家：
日本
起止时间：
1990 至 1991
项目状态：
已结题

项目摘要

Since an initial report by Polge et al. who found a protective action of glycerol in chicken spermatozoa deep-frozen to -79ﾟC, there have been many investigations of freeze-storage of chicken spermatozoa. However, fertility by freeze-thaw semen is markedly below that obtained with fresh semen. The purpose of the present study is to evaluate an effectiveness of trehalose and to improve pelleted methods for cryopreservation of chicken semen. The semen was diluted with an extender containing different levels of trehalose and glycerol, and was frozen in pellets. After thawing, the spem motility and lactate dehydrogenase(LDH)activity of extracellular medium of freeze-thaw semen were measured. Along with the increased levels of trehalose, the motility of postthaw sperm increased and LDH activity in freeze-thaw semen decreased. The minimum difference between LDH leakage before and after freeze-thawing was shown to be at 0.26 M trehalose. The 9% glycerol level gave the best postthaw recovery in the sperm motility and the lowest LDH activity. The effects of PH and osmotic pressure of the extenders containg trehalose on the sperm motility and LDH leakage after freeze-thawing were investigated. The optimum pH value and osmotic pressure for the extender used to freeze semen was 7.2 and 370 mOsm, respectively. Furthermore, the effects of sperm concentration on the sperm motility and LDH leakage after freeze-thawing were estimated. The lowest sperm concentration at freezing gave the best postthaw recovery in the sperm motility and the lowest LDH activity in the extracellular medium of freeze-thaw semen. From those results, it was concluded that cryopreservation of semen diluted with 0.26 M trehalose solution (PH=7.2 ; osmotic pressure, 370 mOsm) containing 9% glycerol was the method feasible for poultry breeding programs.

自Polge等人发现甘油对深度冷冻至-79 C的鸡精子具有保护作用以来，人们对鸡精子的冷冻储存进行了许多研究。然而，冻融精液的生育能力明显低于新鲜精液。本研究的目的是评估海藻糖的有效性，并改进鸡精液的丸剂冷冻保存方法。精液用含有不同水平海藻糖和甘油的扩展剂稀释，并冷冻成颗粒。解冻后，测定冻融精液细胞外培养基中精子活力和乳酸脱氢酶（LDH）活性。随着海藻糖水平的升高，解冻后精子活力增加，冻融后精液LDH活性降低。冻融前后LDH渗漏量的最小差异在海藻糖浓度为0.26 M时。9%甘油水平解冻后精子活力恢复最好，LDH活性最低。研究了海藻糖填充剂的PH和渗透压对冻融后精子活力和LDH泄漏的影响。精液冷冻剂的最佳pH值和渗透压分别为7.2和370 mOsm。分析了精子浓度对冻融后精子活力和LDH渗漏的影响。冷冻时精子浓度最低，解冻后精子活力恢复最好，细胞外培养基中LDH活性最低。由此得出结论，用含有9%甘油的0.26 M海藻糖溶液（PH=7.2，渗透压为370 mOsm）稀释精液进行冷冻保存是可行的家禽育种方案。