性控精液在奶牛胚胎移植产业化中的应用效果研究
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摘要
随着奶牛X、Y精子分离技术的发展,使得受精前控制后代性别成为可能。本实验主要从性控精液与配超排后奶牛和性控胚胎移植妊娠率两方面进行了较系统研究,目的是探讨性控精液在奶牛胚胎移植产业化应用的可行性。
性控精液与配超排奶牛结果:
一、常规冻精组平均回收胚数、平均可用胚数(10.2、6.68)显著高于性控冻精组(6.85、3.74)(P＜0.01)。可用胚胎率常规冻精组显著高于性控冻精组(65.5%vs 54.5%),未受精率显著低于性控冻精组(13.9%vs 21.9%)(P＜0.01)。
二、平均可用胚数性控精液A组显著高于性控冻精B组(5.1vs 2.3)(P＜0.01),平均未受精卵数性控冻精A组显著低于性控精液B组(1.7 vs 2.7)(P＜0.05)。不同输精精剂量(2支vs 3支),可用胚率3支组显著高于2支组(60.5%vs 40.6%)(P＜0.05),未受精卵率3支组显著低于2支组(22.4%vs 39.1%)(P＜0.05);输精剂量(3支vs 4支)受精效果无显著差异,但未受精卵率输精4支组显著低于输精3支组(15.2%vs 30.7%)(P＜0.01)。
三、性控冻精不同输精时间,可用胚率13～15h组(65.6%)显著高于11～13h组和15～17h组(P＜0.05),13～15h组的退化胚率(12.5%)和未受精卵率(21.9%)也低于11～13h组的(22.2%、37.1%)和15～17h组的(27.2%、29.1%),但差异不显著(P＞0.05)。
四、不同输精部位对常规冻精超排受精效果无差异,性控冻精深部输精受精效果好于浅部输精,可用胚率(56.1 vs 46.8)、未受精卵率(22.7%vs 36.7%),但差异不显著(P＞0.05)。
性控胚胎移植结果:
一、性控鲜胚移植妊娠率为48.8%略高于性控冻胚的移植妊娠率44.4%,但两者差异不显著(P＞0.05)。普通胚胎移植妊娠率为51.2%,性控胚胎为46.2%,性鉴胚胎为49.0%,普通胚胎、性控胚胎和性鉴胚胎三者的妊娠率差异不显著(P＞0.05)。
二、普通胚胎、性控胚胎和性鉴胚胎胚胎的产犊率分别为91.2%、91.8%和89.9%,差异不显著(P＞0.05)。母犊率以性鉴胚胎组最高97.9%,但与性控胚胎母犊率91.1%差异不显著(P＞0.05),两者均明显高于常规胚胎组母犊率(48.2%)(p＜0.01)。
三、常规胚胎组、性控胚胎组和性鉴胚胎组的流产或死胎率分别为8.8%、8.2%和10.1%,差异不显著(P＞0.05)。
初步研究表明,性控精液可以作为奶牛胚胎移植产业化的组成部分进行推广,但是还需要对性控精液使用程序和性控胚胎移植所产后代发育情况进一步研究。
It becames possible for determining sex of offsprings pre-fertilization along with development of X and Y sperm sorting technology. This test is designed to study on effect of sexing semen on bovine superovulation and sexing embryo on pregnant rate systemically and discuss the feasibility of application of sexing semen in industrialization of dairy embryo transfer.
The effect of sexing sperm on dairy superovulation indicated:
1 .The total quantity of and viable embryos obtained mated with routine frozen semen were the remarkable higher than ones with sexing sperm, which were 10.2,6.68 vs 6.85, 3.74 and both of them had a remarkable difference(p<0.01). The rate of available embryos and unfertilized oocytes are 65.5%, 13.9% vs 54.5%, 21.9% respectively, which had a remarkable difference (p<0.01), but the rate of degenerated embryos are 20.6% and 23.6%, which had no significant difference. (p>0.05).
2. The study on the different concentration and dosage of sexing semen found that there were no significant difference in the average of embryos collected mated with different concentration of sexing sperm(8.1 vs 6.4), (P>0.05), and got the similar results in the average of total quantity of, viable and degenerated embryos collected mated with 3 or 4 straws(5.94, 2.82, 1.29 vs 7.76, 4.65, 1.94), but there were significant difference in the rate of unfertilized oocytes (30.7% vs 15.2%)collected inseminated with 3 vs 4 straws(p<0.01).
3. The average of embryos collected in the group of 11-13h performing insemination post estrous was remarkable higher than that of 15-17h(4.5 vs 8.0)(P<0.05). The group of 13-15h performing insemination post estrous was remarkable higher than that of 1 l-13h(4.67vs 1.83)(P<0.05), but no siginificant difference comparing with 15-17h groups (P<0.05).
4. There was a better result by performing sexing semen into each horn of uterus in dairy superovulation,there was no significant difference with different part insemination of uterus in routine semen and sexing semen groups, but the average of unfertilized oocytes was significant difference in sexing semen group (P<0.05).
Analysis of effect of sexing embryos on pregnant rate of recipients shows:
1. the pregnant rate of sexing fresh embryos was a little higher than that of sexing frozen embryos (51.2% vs 44.4%),but both of them were no siginificant difference(P>0.05). There were no remarkable difference in pregnant rate of regular embryos(51.2%), sexing embryos(46.2%) and sexed embryos(49.0%).
2.The rate of delivering offsprings origined from regular embryos,sexing embryos and sexed embryos was 91.2%, 91.8%and 89.9% repectively and no siginificant difference above three types of embryos. The rate of delivering female offsprings origined from sexed embryos is the highest(97.9%) in three groups, but no siginificant difference comparing with sexing embryos(p>0.05), of course, both of two groups as above were higher than regular embryos group(p<0.01).
3.The rate of abortion or fetal death origined from regular embryos, sexing embryos and sexed embryos were 8.8%, 8.2% and 10.1%, which no remarkable diffence among them(P>0.05).
In summary, the research of superovulation with sexing sperm and pregnancy rate of sexing embryo transfer indicated that sexing semen as part of industrialization of dairy embryo transfer can extension in the future, but need to research further in the protocol of usage of sexing sperm and development of offsprings origined from sexing embryos.
性控精液与配超排奶牛结果:
一、常规冻精组平均回收胚数、平均可用胚数(10.2、6.68)显著高于性控冻精组(6.85、3.74)(P＜0.01)。可用胚胎率常规冻精组显著高于性控冻精组(65.5%vs 54.5%),未受精率显著低于性控冻精组(13.9%vs 21.9%)(P＜0.01)。
二、平均可用胚数性控精液A组显著高于性控冻精B组(5.1vs 2.3)(P＜0.01),平均未受精卵数性控冻精A组显著低于性控精液B组(1.7 vs 2.7)(P＜0.05)。不同输精精剂量(2支vs 3支),可用胚率3支组显著高于2支组(60.5%vs 40.6%)(P＜0.05),未受精卵率3支组显著低于2支组(22.4%vs 39.1%)(P＜0.05);输精剂量(3支vs 4支)受精效果无显著差异,但未受精卵率输精4支组显著低于输精3支组(15.2%vs 30.7%)(P＜0.01)。
三、性控冻精不同输精时间,可用胚率13～15h组(65.6%)显著高于11～13h组和15～17h组(P＜0.05),13～15h组的退化胚率(12.5%)和未受精卵率(21.9%)也低于11～13h组的(22.2%、37.1%)和15～17h组的(27.2%、29.1%),但差异不显著(P＞0.05)。
四、不同输精部位对常规冻精超排受精效果无差异,性控冻精深部输精受精效果好于浅部输精,可用胚率(56.1 vs 46.8)、未受精卵率(22.7%vs 36.7%),但差异不显著(P＞0.05)。
性控胚胎移植结果:
一、性控鲜胚移植妊娠率为48.8%略高于性控冻胚的移植妊娠率44.4%,但两者差异不显著(P＞0.05)。普通胚胎移植妊娠率为51.2%,性控胚胎为46.2%,性鉴胚胎为49.0%,普通胚胎、性控胚胎和性鉴胚胎三者的妊娠率差异不显著(P＞0.05)。
二、普通胚胎、性控胚胎和性鉴胚胎胚胎的产犊率分别为91.2%、91.8%和89.9%,差异不显著(P＞0.05)。母犊率以性鉴胚胎组最高97.9%,但与性控胚胎母犊率91.1%差异不显著(P＞0.05),两者均明显高于常规胚胎组母犊率(48.2%)(p＜0.01)。
三、常规胚胎组、性控胚胎组和性鉴胚胎组的流产或死胎率分别为8.8%、8.2%和10.1%,差异不显著(P＞0.05)。
初步研究表明,性控精液可以作为奶牛胚胎移植产业化的组成部分进行推广,但是还需要对性控精液使用程序和性控胚胎移植所产后代发育情况进一步研究。
It becames possible for determining sex of offsprings pre-fertilization along with development of X and Y sperm sorting technology. This test is designed to study on effect of sexing semen on bovine superovulation and sexing embryo on pregnant rate systemically and discuss the feasibility of application of sexing semen in industrialization of dairy embryo transfer.
The effect of sexing sperm on dairy superovulation indicated:
1 .The total quantity of and viable embryos obtained mated with routine frozen semen were the remarkable higher than ones with sexing sperm, which were 10.2,6.68 vs 6.85, 3.74 and both of them had a remarkable difference(p<0.01). The rate of available embryos and unfertilized oocytes are 65.5%, 13.9% vs 54.5%, 21.9% respectively, which had a remarkable difference (p<0.01), but the rate of degenerated embryos are 20.6% and 23.6%, which had no significant difference. (p>0.05).
2. The study on the different concentration and dosage of sexing semen found that there were no significant difference in the average of embryos collected mated with different concentration of sexing sperm(8.1 vs 6.4), (P>0.05), and got the similar results in the average of total quantity of, viable and degenerated embryos collected mated with 3 or 4 straws(5.94, 2.82, 1.29 vs 7.76, 4.65, 1.94), but there were significant difference in the rate of unfertilized oocytes (30.7% vs 15.2%)collected inseminated with 3 vs 4 straws(p<0.01).
3. The average of embryos collected in the group of 11-13h performing insemination post estrous was remarkable higher than that of 15-17h(4.5 vs 8.0)(P<0.05). The group of 13-15h performing insemination post estrous was remarkable higher than that of 1 l-13h(4.67vs 1.83)(P<0.05), but no siginificant difference comparing with 15-17h groups (P<0.05).
4. There was a better result by performing sexing semen into each horn of uterus in dairy superovulation,there was no significant difference with different part insemination of uterus in routine semen and sexing semen groups, but the average of unfertilized oocytes was significant difference in sexing semen group (P<0.05).
Analysis of effect of sexing embryos on pregnant rate of recipients shows:
1. the pregnant rate of sexing fresh embryos was a little higher than that of sexing frozen embryos (51.2% vs 44.4%),but both of them were no siginificant difference(P>0.05). There were no remarkable difference in pregnant rate of regular embryos(51.2%), sexing embryos(46.2%) and sexed embryos(49.0%).
2.The rate of delivering offsprings origined from regular embryos,sexing embryos and sexed embryos was 91.2%, 91.8%and 89.9% repectively and no siginificant difference above three types of embryos. The rate of delivering female offsprings origined from sexed embryos is the highest(97.9%) in three groups, but no siginificant difference comparing with sexing embryos(p>0.05), of course, both of two groups as above were higher than regular embryos group(p<0.01).
3.The rate of abortion or fetal death origined from regular embryos, sexing embryos and sexed embryos were 8.8%, 8.2% and 10.1%, which no remarkable diffence among them(P>0.05).
In summary, the research of superovulation with sexing sperm and pregnancy rate of sexing embryo transfer indicated that sexing semen as part of industrialization of dairy embryo transfer can extension in the future, but need to research further in the protocol of usage of sexing sperm and development of offsprings origined from sexing embryos.
引文
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