添加低剂量大豆卵磷脂稀释液低温保存绵羊精液的优化研究
|
摘要
本实验旨在探讨添加低剂量大豆卵磷脂稀释液对绵羊精液低温保存效果。实验一采用6个浓度(0%、0.25%、0.5%、0.75%、1.0%、1.25%)大豆卵磷脂替代TRIS专利稀释液中卵黄低温保存绵羊精液,在第0、1、4、7、10、12、18天检测精子活率、顶体完整率;实验二选用实验一中最优添加组(0.5%组)和TRIS专利稀释液制成粉剂低温保存绵羊精液,在保存第0、1、4、7、10、12天对精子活率和顶体完整率进行测定,并在保存第10天对绵羊进行人工授精。结果表明:实验一中,0%组从第1天精子活率低于其他组(P<0.05),0.25%组精子活率观测值从第10天开始低于0.25%以上浓度组(P<0.05),0.5%、0.75%、1.0%、1.25%组保存第10天精子活率均大于50%,0.5%组保存第18天精子活率高于1.0%、1.25%组(P<0.05);各组顶体完整率缓慢下降,各时间点均无显著性差异;实验二中,0.5%组与TRIS组在各时间点的精子活率、顶体完整率与受胎率均无显著性差异,保存第10天精子活率均高于50%;0.5%组受胎率为65.49%,略低于TRIS组67.65%(P>0.05)。本实验条件下,绵羊精液低温保存稀释液中添加0.5%大豆卵磷脂替代卵黄效果最佳。
The aim of this experiment was to investigate the effect of low-dose soybean lecithin diluent on sheep semen cryopreservation. In experiment 1, six concentrations(0%, 0.25%, 0.5%, 0.75%, 1.0%, 1.25%)of soybean lecithin instead of egg yolk in patent dilution of TRIS to preserve sheep semen at low temperature. Sperm motility and acrosomal integrity rate were detected at 0, 1, 4, 7, 10, 12 and 18 day. In the experiment 2, the best addition group(0.5%) and TRIS patent diluent were used to make powder for cryopreservation of sheep semen, sperm motility and acrosomal integrity rate were detected at 0,1, 4, 7, 10, 12 day, and artificial insemination was performed on sheep semen on the 10 th day after storage. The results showed that the sperm motility of 0% group from the first day was significantly lower than that of other groups(P<0.05). the sperm motility in 0.25% group was significantly lower than that in above 0.25% concentration group from the 10 th day P<0.05), The sperm motility of 0.5%, 0.75%, 1%, 1.25% group on the 10 th day was higher than 50%. The sperm motility in 0.5% group was significantly higher than that in 1.0%, 1.25% group on the 18 th day(P<0.05). There was no significant difference in acrosomal integrity at all time points. There was no significant difference between 0.5% group and TRIS group at each time point. The sperm motility rate was higher than that of 50% 0.5% group on the 10 th day(P>0.05). The conception rate of 0.5% group was slightly lower than that of TRIS group 67.65%(P>0.05). Adding 0.5% soybean lecithin to the diluent of sheep semen cryopreservation is the best alternative to egg yolk.
The aim of this experiment was to investigate the effect of low-dose soybean lecithin diluent on sheep semen cryopreservation. In experiment 1, six concentrations(0%, 0.25%, 0.5%, 0.75%, 1.0%, 1.25%)of soybean lecithin instead of egg yolk in patent dilution of TRIS to preserve sheep semen at low temperature. Sperm motility and acrosomal integrity rate were detected at 0, 1, 4, 7, 10, 12 and 18 day. In the experiment 2, the best addition group(0.5%) and TRIS patent diluent were used to make powder for cryopreservation of sheep semen, sperm motility and acrosomal integrity rate were detected at 0,1, 4, 7, 10, 12 day, and artificial insemination was performed on sheep semen on the 10 th day after storage. The results showed that the sperm motility of 0% group from the first day was significantly lower than that of other groups(P<0.05). the sperm motility in 0.25% group was significantly lower than that in above 0.25% concentration group from the 10 th day P<0.05), The sperm motility of 0.5%, 0.75%, 1%, 1.25% group on the 10 th day was higher than 50%. The sperm motility in 0.5% group was significantly higher than that in 1.0%, 1.25% group on the 18 th day(P<0.05). There was no significant difference in acrosomal integrity at all time points. There was no significant difference between 0.5% group and TRIS group at each time point. The sperm motility rate was higher than that of 50% 0.5% group on the 10 th day(P>0.05). The conception rate of 0.5% group was slightly lower than that of TRIS group 67.65%(P>0.05). Adding 0.5% soybean lecithin to the diluent of sheep semen cryopreservation is the best alternative to egg yolk.
引文
[1]De Paz P,Esteso M C,Alvarez M,et al.Development of extender based on soybean lecithin for its application in liquid ram semen[J].Theriogenology,2010,74(4):663-671.
[2]Ramanathan K,Vanmathy K,Ahmed T,et al.Effect of semen extenders on sperm parameters of ram semen during liquid storage at 4℃[J].Small Ruminant Res,2011,99:208-213.
[3]Beccaglia M,Anastasi P,Chigioni S,et al.Tris-lecithin extender supplemented with antioxidant catalase for chilling of canine semen[J].Reprod Domest Anim,2009,44(2):345-349.
[4]Kmenta I,Strohmayer C,Müller-Schl?sser F,et al.Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of coldstored canine spermatozoa[J].Theriogenology,2011,75:1095-1103.
[5]赵建清,王杰,李娜,等.大豆卵磷脂替代卵黄的稀释液低温保存绵羊精液效果[J].塔里木大学学报,2018(4):15-19.
[6]王杰,李娜,肖国亮,等.稀释液冻干粉剂低温超长时间保存绵羊精液效果[J].中国草食动物科学,2018,38(3):64-66.
[7]Forouzanfara M,Sharafi M,Hosseini S M,et al.In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen[J].Theriogenology,2010,73(4):480-487.
[8]孟娜娜,孙树春,王寒阳,等.绵羊精液4℃液态保存的稀释液配方优化研究[J].中国畜牧杂志,2015,51(5):34-38.
[9]张树棣.不同组分精液稀释液对牛精子冷冻保护效果的研究[D].呼和浩特:内蒙古农业大学,2007.
[10]曹宗顺,卢凤琦.磷脂的提取与精制[J].中国医药工业杂志,1995,26(l):42-43.
[11]Witte T S,Schafer-Somi S,Kuchar A,et al.Effect of hens egg yolk on capacitation and acrosome reaction of diluted canine spermatozoa[J].Anim Reprod Sci,2009,110:293-305.
[12]Ashworth P J,Harrison R A,Miller N G,et al.Survival of ram spermatozoa at high dilutions:protective effect of simple constituents of culture media as compared with seminal plasma[J].Reprod Fertil Dev,1994,6:173-180.
[13]迟玉杰.蛋制品加工技术[M].北京:中国轻工业出版社,2009.
[14]陈晓利,郑永富,王杰,等.超长时间低温保存绵羊鲜精活率和DNA完整性的比较[J].黑江畜牧兽医,2017(20):90-92.
[15]费俊阳,高庆华,肖国亮.绵羊鲜精超长时间保存技术研究[J].黑龙江畜牧兽医,2018(14):62-65.
[16]覃能斌,李孝娟.浅谈家畜人工授精发展[J].黑龙江动物繁殖,2017,25(6):17-19.
[2]Ramanathan K,Vanmathy K,Ahmed T,et al.Effect of semen extenders on sperm parameters of ram semen during liquid storage at 4℃[J].Small Ruminant Res,2011,99:208-213.
[3]Beccaglia M,Anastasi P,Chigioni S,et al.Tris-lecithin extender supplemented with antioxidant catalase for chilling of canine semen[J].Reprod Domest Anim,2009,44(2):345-349.
[4]Kmenta I,Strohmayer C,Müller-Schl?sser F,et al.Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of coldstored canine spermatozoa[J].Theriogenology,2011,75:1095-1103.
[5]赵建清,王杰,李娜,等.大豆卵磷脂替代卵黄的稀释液低温保存绵羊精液效果[J].塔里木大学学报,2018(4):15-19.
[6]王杰,李娜,肖国亮,等.稀释液冻干粉剂低温超长时间保存绵羊精液效果[J].中国草食动物科学,2018,38(3):64-66.
[7]Forouzanfara M,Sharafi M,Hosseini S M,et al.In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen[J].Theriogenology,2010,73(4):480-487.
[8]孟娜娜,孙树春,王寒阳,等.绵羊精液4℃液态保存的稀释液配方优化研究[J].中国畜牧杂志,2015,51(5):34-38.
[9]张树棣.不同组分精液稀释液对牛精子冷冻保护效果的研究[D].呼和浩特:内蒙古农业大学,2007.
[10]曹宗顺,卢凤琦.磷脂的提取与精制[J].中国医药工业杂志,1995,26(l):42-43.
[11]Witte T S,Schafer-Somi S,Kuchar A,et al.Effect of hens egg yolk on capacitation and acrosome reaction of diluted canine spermatozoa[J].Anim Reprod Sci,2009,110:293-305.
[12]Ashworth P J,Harrison R A,Miller N G,et al.Survival of ram spermatozoa at high dilutions:protective effect of simple constituents of culture media as compared with seminal plasma[J].Reprod Fertil Dev,1994,6:173-180.
[13]迟玉杰.蛋制品加工技术[M].北京:中国轻工业出版社,2009.
[14]陈晓利,郑永富,王杰,等.超长时间低温保存绵羊鲜精活率和DNA完整性的比较[J].黑江畜牧兽医,2017(20):90-92.
[15]费俊阳,高庆华,肖国亮.绵羊鲜精超长时间保存技术研究[J].黑龙江畜牧兽医,2018(14):62-65.
[16]覃能斌,李孝娟.浅谈家畜人工授精发展[J].黑龙江动物繁殖,2017,25(6):17-19.