冷冻稀释液中添加大豆卵磷脂对梅花鹿冻融精子质量的影响

doi:10.16431/j.cnki.1671-7236.2022.11.023

摘要/Abstract

摘要： 【目的】探究在冷冻稀释液中添加大豆卵磷脂代替10%卵黄对梅花鹿精液冷冻保存效果的影响，为梅花鹿人工授精体系的完善提供参考。【方法】采用电刺激法采集梅花鹿精液，以精液冷冻稀释液中分别添加1%、2%、3%、4%和5%大豆卵磷脂代替10%卵黄作为试验组，添加20%卵黄作为对照组，分别进行各组精液冷冻保存。5 d后，进行精液解冻，检测解冻后各组精子的活力、质膜完整率、顶体完整率、线粒体活性、存活时间，筛选合适浓度的大豆卵磷脂。选取4~5岁健康雌性梅花鹿，肌肉注射300 IU孕马血清促性腺激素（PMSG）和0.4 mg氯前列醇钠进行同期发情处理，发情后第20 h用20%卵黄组与筛选出的大豆卵磷脂组冻精进行人工输精，输精后30 d使用B超检测仪检测妊娠情况，统计妊娠率。【结果】与对照组相比，1%大豆卵磷脂组冻融后的精子活力、向前活动力、快速前进活力、活率、质膜完整率、顶体完整率及线粒体活性均显著提高（P<0.05）；随着稀释液中大豆卵磷脂浓度的增加，其冻融后精子活力、向前活动力、快速前进活力、活率、质膜完整率、顶体完整率以及线粒体活性呈下降趋势，精子存活时间也随浓度的增加而减少。1%大豆卵磷脂组冻融精子人工授精梅花鹿的妊娠率为61.11%，高于对照组、2%和3%大豆卵磷脂组，但差异均不显著（P>0.05）。【结论】在梅花鹿精子冷冻稀释液中添加1%大豆卵磷脂替代10%卵黄，能有效提高梅花鹿冻融精子的质量，为进一步筛选新型梅花鹿精液冷冻稀释液提供理论基础。

关键词: 梅花鹿; 大豆卵磷脂; 精子; 冷冻保存; 卵黄

Abstract: 【Objective】 The aim of this study was to explore the effects of adding soybean lecithin instead of 10% egg yolk to the frozen diluent on the cryopreservation of semen of sika deer, in order to provide references for the improvement of the artificial insemination system of sika deer.【Method】 The semen of sika deer was collected by electrical stimulation, and 1%, 2%, 3%, 4% and 5% soybean lecithin was added to the frozen diluent to replace 10% egg yolk as the experimental group, and 20% egg yolk was added as control group, respectively.Cryopreservation of semen in each group was performed.After 5 days, the semen was thawed, and the sperm motility, plasma membrane integrity rate, acrosome integrity rate, mitochondrial activity, and survival time of each group were detected after thawing, and soybean lecithin of suitable concentration was screened.Healthy female sika deer aged 4-5 years were selected, intramuscularly injected with 300 IU PMSG and 0.4 mg cloprostol sodium for simultaneous estrus treatment, and artificial insemination was performed with frozen semen in the 20% egg yolk group and the selected soybean lecithin group at 20 h after estrus, 30 days after insemination, B-ultrasound detector was used to detect the pregnancy, and count the pregnancy rate.【Result】 Compared with control group, the sperm motility, forward motility, fast-forward motility, motility, plasma membrane integrity rate, acrosome integrity rate and mitochondrial activity were significantly improved in 1% soybean lecithin group after freezing and thawing (P<0.05).With the increase of the concentration of soybean lecithin in the diluent, the sperm motility, forward motility, fast forward motility, motility, plasma membrane integrity rate, acrosome integrity rate and mitochondrial activity showed a downward trend, and sperm survival time also decreased with the increasing concentration.The pregnancy rate of sika deer in 1% soybean lecithin group was 61.11% after artificial insemination of frozen-thawed sperm, which was higher than that of control group, 2% and 3% soybean lecithin groups, but the difference was not significant (P>0.05).【Conclusion】 Adding 1% soybean lecithin to replace 10% egg yolk in the frozen diluent of sika deer sperm could effectively improve the quality of the frozen and thawed semen of sika deer, and provide a theoretical basis for further screening of new frozen diluents of sika deer semen.

Key words: sika deer; soybean lecithin; sperm; cryopreservation; egg yolk

中图分类号:

S825

李梦杰, 赵伟刚, 郭肖兰, 顾士钢, 赵全民, 徐超. 冷冻稀释液中添加大豆卵磷脂对梅花鹿冻融精子质量的影响[J]. 中国畜牧兽医, 2022, 49(11): 4327-4334.

LI Mengjie, ZHAO Weigang, GUO Xiaolan, GU Shigang, ZHAO Quanmin, XU Chao. Effects of Soybean Lecithin in Frozen Diluent on Quality of Frozen-thawed Sperm of Sika Deer[J]. China Animal Husbandry and Veterinary Medicine, 2022, 49(11): 4327-4334.

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