脱脂奶粉和大豆卵磷脂对绵羊精液低温保存效果的影响
发布时间：2018-09-05 20:51
【摘要】：本试验探讨了添加脱脂奶粉替代鲜牛奶和大豆卵磷脂替代卵黄在精液低温保存的可行性,旨在开发适宜在养羊研究及生产实践中应用的绵羊精液低温保存稀释液。假阴道法采集绵羊精液,用不同浓度的脱脂奶粉和大豆卵磷脂分别替代绵羊精液稀释液中的鲜牛奶和卵黄,对绵羊精液稀释并进行低温保存,定时复温并对绵羊精子的活力、存活时间、活精子百分率、畸形率和顶体完整率进行检测,从而评价绵羊精液的低温保存效果,并将低温保存稀释液用于成年绵羊体外受精试验进行验证。在绵羊精液稀释中添加不同浓度的(4%、7%、10%、13%)脱脂奶粉替代鲜牛奶作为试验组,对照组则添加牛奶,试验结果表明:精子活力变化情况,在0~60h各脱脂奶粉组稀释液与牛奶组无显著差异(P0.05),但10%脱脂奶粉组高于其他脱脂奶粉组,在72~84h时,10%脱脂奶粉组与牛奶组无显著性差异(P0.05),而4%、13%脱脂奶粉组显著低于牛奶组(P0.05),在72~132h,其他脱脂奶粉组(4%、7%、13%)均低于10%脱脂奶粉组和牛奶组(P0.05);活精子百分率变化情况,在0~72h各个脱脂奶粉组均与牛奶组无显著性差异(P0.05),到96h,10%脱脂奶粉组最高,达到62.62%,与牛奶组差异不显著(P0.05),120h时,10%脱脂奶粉组与牛奶组无显著性差异(P0.05),但是显著高于其他脱脂奶粉组(P0.05);精子畸形率变化情况,在24h、72h和96h三个时间点,7%、10%脱脂奶粉组与牛奶组相互之间差异不显著(P0.05),10%脱脂奶粉组畸形率最低,在120h时,10%脱脂奶粉组与牛奶组差异不显著(P0.05),并显著低于其他脱脂奶粉组(P0.05);精子顶体完整率变化情况,在48h时10%脱脂奶粉组与牛奶组差异不显著(P0.05),显著高于4%和13%脱脂奶粉组(P0.05)。综合以上各指标检测结果,以10%脱脂奶粉替代鲜牛奶的效果为最佳。在以10%脱脂奶粉替代鲜牛奶进行优化绵羊低温精液稀释液配方的基础上,利用不同浓度(0.375%、0.750%、1.500%、3.000%)的大豆卵磷脂取代传统稀释液中的卵黄,试验结果表明:精子活力变化情况,在0~120h各时间点,0.375%大豆卵磷脂组精子活力与卵黄组差异均不显著(P0.05),其中在84h时,0.375%大豆卵磷脂组精子活力还维持在0.43,在60~96h,1.500%、3.000%大豆卵磷脂组活力均显著低于卵黄组和0.375%大豆卵磷脂组(P0.05),在108h时,0.750%、1.500%、3.000%大豆卵磷脂组活力均显著低于卵黄组(P0.05);精子存活时间情况,0.375%大豆卵磷脂的有效存活时间为88h,总存活时间为120h,与卵黄组组差异不显著(P0.05),其余各大豆卵磷脂组的有效存活时间和总存活时间均显著低于0.375%大豆卵磷脂组和卵黄组(P0.05);活精子百分率变化情况,在0、24h时间点,各个组间无显著性差异(P0.05),从48h~120h,0.375%大豆卵磷脂组与卵黄组差异不显著(P0.05),尤其在72h、96h、120h,0.375%大豆卵磷脂组与卵黄组差异不显著(P0.05),但显著高于0.750%、1.500%、3.000%大豆卵磷脂组(P0.05);精子畸形率变化情况,0~120h,0.375%大豆卵磷脂组与卵黄组差异均不显著(P0.05),在72h、96h时,0.375%大豆卵磷脂组畸形率最低;精子顶体完整率变化情况,0~120h,各个大豆卵磷脂组均与卵黄组差异不显著(P0.05)。综合以上结果可以看出,0.375%的大豆卵磷脂组替代卵黄效果最好。利用最优化的绵羊精液低温保存稀释液(10%脱脂奶粉替代新鲜牛奶,0.375%大豆卵磷脂替代卵黄)低温保存绵羊精精液,进行绵羊的体外受精试验,以冷冻精液做为对照,结果表明:低温组精液的绵羊卵母细胞体外受精卵裂率与冷冻组无显著性差异(P0.05),但低温组卵裂率(63.12%)高于冷冻组(55.92%),低温组的囊胚发育率(20.61%)显著高于冷冻组(14.27%)(P0.05);囊胚经荧光染色后计数细胞数,低温组(111.64)和冷冻组(109.60)无显著差异(P0.05)。
[Abstract]:The feasibility of using skimmed milk powder instead of fresh milk and soybean lecithin instead of yolk in cryopreservation of semen was studied in this experiment. The purpose of this experiment was to develop Sheep Semen Cryopreservation dilution suitable for sheep breeding research and production practice. The fresh milk and egg yolk in sheep semen dilution solution were diluted and cryopreserved, and the sperm viability, survival time, percentage of live sperm, deformity rate and acrosome integrity of sheep were detected. The cryopreserved semen dilution was used in adult sheep. External fertilization test was carried out to verify the results. Different concentrations of skimmed milk powder (4%, 7%, 10%, 13%) were added in the dilution of Sheep Semen to replace fresh milk as the experimental group, while milk was added in the control group. The results showed that there was no significant difference in sperm motility between the skimmed milk powder group and the milk group at 0-60 h (P 0.05), but the 10% skimmed milk powder group was higher than the milk group. Other skimmed milk powder groups, at 72-84 hours, 10% skimmed milk powder group and milk group had no significant difference (P 0.05), but 4%, 13% skimmed milk powder group was significantly lower than milk group (P 0.05), at 72-132 hours, other skimmed milk powder group (4%, 7%, 13%) were lower than 10% skimmed milk powder group and milk group (P 0.05); the percentage of live sperm was changed in each skimmed milk powder group at 0-72 hours. There was no significant difference between the milk group and the 10% skimmed milk powder group (P 0.05). At 96 h, the 10% skimmed milk powder group was the highest, reaching 62.62%, which was not significantly different from the milk group (P 0.05). At 120 h, the 10% skimmed milk powder group and the milk group had no significant difference (P 0.05), but significantly higher than the other skimmed milk powder group (P 0.05); the change of sperm deformity rate was 7%, 10% skimmed at 24 h, 72 h and 96 h. There was no significant difference between the fat milk powder group and the milk group (P 0.05). The deformity rate of the 10% skimmed milk powder group was the lowest. At 120 hours, the deformity rate of the 10% skimmed milk powder group and the milk group was not significantly different (P 0.05), and was significantly lower than that of the other skimmed milk powder groups (P 0.05). It was significantly higher than that of 4% and 13% skimmed milk powder group (P 0.05). According to the above results, 10% skimmed milk powder was the best substitute for fresh milk. The results showed that the sperm motility of 0.375% soybean lecithin group was not significantly different from that of egg yolk group (P 0.05) at 0-120 H. The sperm motility of 0.375% soybean lecithin group remained at 0.43, 60-96 h, 1.500%, 3.000% soybean lecithin group was significantly lower than that of egg at 84 H. The activity of 0.750%, 1.500% and 3.000% soybean lecithin group was significantly lower than that of lecithin group (P 0.05) at 108 hours in yellow group and 0.375% soybean lecithin group (P 0.05); the effective survival time of sperm in 0.375% soybean lecithin group was 88 hours and the total survival time was 120 hours, which was not significantly different from that in yolk group (P 0.05). Survival time and total survival time were significantly lower than 0.375% soybean lecithin group and yolk group (P 0.05); the percentage of live sperm had no significant difference (P 0.05) between each group at 0 and 24 hours, and there was no significant difference (P 0.05) between soybean lecithin group and yolk group from 48 to 120 hours, 0.375% soybean lecithin group and yolk group (P 0.05), especially at 72 hours, 96 hours, 120 hours, 0.375% soybean lecithin group and egg yolk group. There was no significant difference between yellow group and yolk group (P 0.05), but it was significantly higher than 0.750%, 1.500%, 3.000% soybean lecithin group (P 0.05); sperm deformity rate was not significantly different between 0-120 h, 0.375% soybean lecithin group and yolk group (P 0.05); at 72 h, 96 h, 0.375% soybean lecithin group had the lowest deformity rate; acrosome integrity rate of sperm, 0-120 h, each soybean egg. The results showed that 0.375% soybean lecithin group had the best effect on replacing yolk. Sheep semen was cryopreserved in vitro with the optimum dilution of Sheep Semen (10% skim milk instead of fresh milk, 0.375% soybean lecithin instead of yolk). The results showed that there was no significant difference in cleavage rate between cryogenic group and cryogenic group (P 0.05), but the cleavage rate of cryogenic group (63.12%) was higher than that of cryogenic group (55.92%). The blastocyst development rate of cryogenic group (20.61%) was significantly higher than that of cryogenic group (14.27%) (P 0.05). The number of counted cells in cryogenic group (111.64) and cryopreservation group (109.60) had no significant difference (P0.05).
【学位授予单位】：河北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826
本文编号：2225429
[Abstract]:The feasibility of using skimmed milk powder instead of fresh milk and soybean lecithin instead of yolk in cryopreservation of semen was studied in this experiment. The purpose of this experiment was to develop Sheep Semen Cryopreservation dilution suitable for sheep breeding research and production practice. The fresh milk and egg yolk in sheep semen dilution solution were diluted and cryopreserved, and the sperm viability, survival time, percentage of live sperm, deformity rate and acrosome integrity of sheep were detected. The cryopreserved semen dilution was used in adult sheep. External fertilization test was carried out to verify the results. Different concentrations of skimmed milk powder (4%, 7%, 10%, 13%) were added in the dilution of Sheep Semen to replace fresh milk as the experimental group, while milk was added in the control group. The results showed that there was no significant difference in sperm motility between the skimmed milk powder group and the milk group at 0-60 h (P 0.05), but the 10% skimmed milk powder group was higher than the milk group. Other skimmed milk powder groups, at 72-84 hours, 10% skimmed milk powder group and milk group had no significant difference (P 0.05), but 4%, 13% skimmed milk powder group was significantly lower than milk group (P 0.05), at 72-132 hours, other skimmed milk powder group (4%, 7%, 13%) were lower than 10% skimmed milk powder group and milk group (P 0.05); the percentage of live sperm was changed in each skimmed milk powder group at 0-72 hours. There was no significant difference between the milk group and the 10% skimmed milk powder group (P 0.05). At 96 h, the 10% skimmed milk powder group was the highest, reaching 62.62%, which was not significantly different from the milk group (P 0.05). At 120 h, the 10% skimmed milk powder group and the milk group had no significant difference (P 0.05), but significantly higher than the other skimmed milk powder group (P 0.05); the change of sperm deformity rate was 7%, 10% skimmed at 24 h, 72 h and 96 h. There was no significant difference between the fat milk powder group and the milk group (P 0.05). The deformity rate of the 10% skimmed milk powder group was the lowest. At 120 hours, the deformity rate of the 10% skimmed milk powder group and the milk group was not significantly different (P 0.05), and was significantly lower than that of the other skimmed milk powder groups (P 0.05). It was significantly higher than that of 4% and 13% skimmed milk powder group (P 0.05). According to the above results, 10% skimmed milk powder was the best substitute for fresh milk. The results showed that the sperm motility of 0.375% soybean lecithin group was not significantly different from that of egg yolk group (P 0.05) at 0-120 H. The sperm motility of 0.375% soybean lecithin group remained at 0.43, 60-96 h, 1.500%, 3.000% soybean lecithin group was significantly lower than that of egg at 84 H. The activity of 0.750%, 1.500% and 3.000% soybean lecithin group was significantly lower than that of lecithin group (P 0.05) at 108 hours in yellow group and 0.375% soybean lecithin group (P 0.05); the effective survival time of sperm in 0.375% soybean lecithin group was 88 hours and the total survival time was 120 hours, which was not significantly different from that in yolk group (P 0.05). Survival time and total survival time were significantly lower than 0.375% soybean lecithin group and yolk group (P 0.05); the percentage of live sperm had no significant difference (P 0.05) between each group at 0 and 24 hours, and there was no significant difference (P 0.05) between soybean lecithin group and yolk group from 48 to 120 hours, 0.375% soybean lecithin group and yolk group (P 0.05), especially at 72 hours, 96 hours, 120 hours, 0.375% soybean lecithin group and egg yolk group. There was no significant difference between yellow group and yolk group (P 0.05), but it was significantly higher than 0.750%, 1.500%, 3.000% soybean lecithin group (P 0.05); sperm deformity rate was not significantly different between 0-120 h, 0.375% soybean lecithin group and yolk group (P 0.05); at 72 h, 96 h, 0.375% soybean lecithin group had the lowest deformity rate; acrosome integrity rate of sperm, 0-120 h, each soybean egg. The results showed that 0.375% soybean lecithin group had the best effect on replacing yolk. Sheep semen was cryopreserved in vitro with the optimum dilution of Sheep Semen (10% skim milk instead of fresh milk, 0.375% soybean lecithin instead of yolk). The results showed that there was no significant difference in cleavage rate between cryogenic group and cryogenic group (P 0.05), but the cleavage rate of cryogenic group (63.12%) was higher than that of cryogenic group (55.92%). The blastocyst development rate of cryogenic group (20.61%) was significantly higher than that of cryogenic group (14.27%) (P 0.05). The number of counted cells in cryogenic group (111.64) and cryopreservation group (109.60) had no significant difference (P0.05).
【学位授予单位】：河北农业大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：S826
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