中文摘要：

以伊红染色法检测样品精子存活率为依据，研究了4℃下5种保存液及Ca^2＋、Mg^2＋离子对中华绒螯蟹精子体外保存效果的影响。5种保存液分别为人工海水（ASW）、2倍钙离子人工海水（2×Ca^2＋-一ASW）、无镁离子人工海水（Mg^2＋-FASW）、无钾离子人工海水（K^＋-FASW）、无钙离子人工海水（Ca^2＋-FASW），经4天保存后，各保存液中精子样品的存活率和精子密度均出现明显差异，K^＋-FASW、ASW及2×Ca^2＋-ASW三种保存液中的精子因发生顶体反应而大量死亡，而Mg^2＋-FASW、Ca^2＋-FASW的保存效果较好。在此基础上，进一步探讨了不同Mg^2＋和Ca^2＋浓度对精子存活率的影响，结果发现，经24h保存后各实验组精子存活率均随着两种离子浓度的增加而明显下降。上述结果表明：K^＋-FASW、ASW及2×CaH—ASW不适合精子保存，而Mg^2＋-FASW和Ca^2＋-FASW均可作为该蟹精子的保存液；Ca^2＋因可引起精子顶体反应而造成保存液中精子的大量死亡，其浓度与存活率呈明显的负相关；无K^＋的保存液中，Ca^2＋、Mg^2＋的存在与否对精子的保存效果起关键作用；无Mg^2＋人工海水之所以具有较好的保存效果，可能与Mg^2＋的缺乏而导致Ca^2＋逆浓度差转运受阻，避免了因Ca^2＋进入而诱发顶体反应有关。

英文摘要：

The viability rate of in vitro sperms of Eriocheir sinensis preserved in five solutions at 4℃ was studied with eosin stain to evaluate the impact from metal ions. The five preservation solutions （Tab. 1） included artificial seawater （ASW）, 2-fold calcium ion artificial seawater （2× Ca^2＋-ASW）, magnesium-free artificial seawater （Mg^2＋-FASW）, potassium-free artificial seawater （K^＋-FASW） and calcium-free artificial seawater （Ca^2＋-FASW）. Results show that the sperms preserved in K^＋-FASW, ASW, and 2 × Ca^2＋-ASW were completely killed 4 days later because of acrosome reaction. However, most of the sperms in Mg^2＋-FASW and Ca^2＋-FASW remained alive in the same period. Therefore, Mg^2＋-FASW and Ca^2＋-FASW can be ideal solutions for preserving E.sinensis sperms in vitro, but K^＋-FASW, ASW, and 2 × Ca^2＋-ASW. Furthermore, the viability rate was also checked with different concentrations of Ca^2＋ and Mg^2＋. As the concentrations of Ca^2＋ and Mg^2＋ incremented, the viability rate dropped in 24 hour, indicating that Ca^2＋ could lead to mass mortality of the sperms due to acrosome reaction. Inverse correlation was found between the viability rate and Ca^2＋ concentration. Ca^2＋ and Mg^2＋ played a key role in e K^＋-free preservation solutions. The good performance of Mg^2＋-FASW is probably due to the prohibition of Ca^2＋ transportation against the concentration difference, so that Ca^2＋ was kept away from entering the sperms resulting in acrosome reaction.