山羊胚胎移植效果的影响因素研究
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摘要
近年来，胚胎移植技术在短期内扩大优良种用山羊的数量方面发挥了重要作用。但在实践中，胚胎移植的成功率仍不稳定。国内外已有不少学者在提高胚胎移植成功率方面进行了大量研究，但是这些研究大多是针对个别影响因素的探讨。对影响胚胎移植技术的各环节的因素进行全面的分析将有助于进一步提高其成功率，增加成功率的稳定性。本试验对30只波尔山羊进行超排；89只鲁北白山羊作为受体，针对影响超数排卵、同期发情和受体妊娠率的多种因素进行综合分析。
采用适合性检验对5个超排指标进行正态性分析，结果表明：黄体数、可用胚数、采胚数均服从正态分布。而变性胚数和未受精卵数不服从正态分布，对其应采用非参数检验的方法；多变量方差分析对影响超排效果的6个因素的分析结果表明：FSH的注射时间和同期处理方法在综合效应模型中是产生超排效果差异的主要原因。各因素间的交互作用差异不显著。
采用单因素方差分析和非参数检验对影响超排效果的6个因素进行分析。结果表明：膘情因素四个水平中，中上、中等体况个体的黄体数为(11.17±3.29)枚、(6.22±2.37)枚，显著高于其它水平(P＜0.01)，而变性胚率和未受精卵率居较低水平。说明不同膘情对超排效果的影响主要体现在黄体数方面；三种FSH注射时间以第7d或第9d开始注射的超排效果好，说明波尔山羊的卵泡生长波的启动应该集中在第7～9d；不同的同期处理方法对黄体数量产生极显著差异(P＜0.01)，一次肌注PG2mL剂量过大，会严重影响供体的受精卵率和可用胚数。放栓组和PG注射组比较无显著差异(P＞0.05)。结果表明，PG分两次注射(1mL／次)的总体效果较好；注射LH后其采胚率(29.56％＜63.58％)、可用胚率(75.30％＜94.34％)低于不注射组，并且未受精卵率显著高于不注射组(20.07％＞1.37％)。说明LH的注射增加了不成熟卵泡的排卵数目；同期处理后24h发情的供体在采胚数(6.29±1.57)和可用胚数(5.10±1.56)方面高于36h、42h发情组，同时未受精率也较高(15.90％)，三个组对超排效果的影响差异不显著(P＞0.05)，说明供体发情出现的时间对超排效果的影响并不明显；人工+本交的交配方式可显著降低未受精卵率(P＜0.05)，提高超排效果。
PG的应用可使供体的36h同期发情率达到100％，各水平间以两次注射PG的效果为好，24h发情率达到80％；黄体数与采胚数的相关系数为0.415(P＜0.05)，采胚数和可用胚数的相关系数为0.928(P＜0.01)，在实际应用中可以通过黄体数判断其采胚数和可用胚数。
采用Logistic回归分析了影响受体妊娠率的4种因素，结果表明，移植方式和发情状况对妊娠结果的产生具有决定性作用(P＜0.01)，可通过建立的回归模型对受体妊娠情况进行预测。胚胎发育阶段的差异会极显著影响返情状况；自然发情的受体有较高的不返情率(69.44％)，显著高于诱导发情组(P＜0.05)；受体黄体数量的差异对返情率的影响不显著(P＞0.05)；双侧移植方式可显著提高受体的不返情率(P＜0.01)，达到了80％；桑椹胚的返情率最低，显著低于其它水平(P＜0.01)，达到31.58％。
综合以上研究结果，可以得出如下结论
门）FSH的注射时间和同期处理方法在综合效应模型中是产生超排效果差异的
主要原因。
② 采用如下方法可得到较好的超排效果：选择中上、中等体况的个体作为超
排个体，在第7 d或第9 d采用递减法注射FSH（16 nL），比例为S：5：3，每大2
次，间隔12小时；在超排第3 d上、下午各肌注lmLPG（间隔12 h）；选择同期处
理后 24 h发情的个体；采用本交十人工授精的交配方式；在发情羊配种后 6～7大，
采用于宫角冲胚法可得到较好的超排效果。
（3）采用如下方法可得到最好的妊娠效果：将采集到的桑棋胚移植到同期发情
（同步差为上12 h）的受体内。受体最好为自然发情：采用于宫角双侧移植的方法。
。＿。，、，。，、＿．．＿＿＿，，、、＿＿。、，，＿，＿、＿＿＿C
（4）用发情状况（XI）和移植方式（x；）建立的回归方程P＝＿
l＋e
可对受体妊娠率进行预测，预测准确率为79 3％。
In recent years, embryo transfer technology has made great contributions to increasing the amount of goats for breeding in a short period, but the success rate of embryo transfer was not stable in practice. Many people from native or abroad have carried out lots of study on improving the rate of success, but most of them discussed only about one factor affecting the rate of success. A comprehensive study on the influencial factors in the course of embryo transfer will help to improve the rate of success and enhance the stability of it. In this study, 30 Boer goats as donor; 89 LUBei white goats as recipients of estrus synchronization, analysis on varieties of factors affecting the superovulation, synchronization and the rate of pregnancy.
Analysis on 5 kinds of targets for superovulation by normal test showed: CL (corpus luteum) number, embryo number, usable embryo number were normal, while metamorphic embryo number and no zygote number were unnormal. So nonparametric test method should be adopted for the later; Multivariate Analysis on 6 kinds of influential factors for the superovulation effect showed: The injection time of FSH and estrous synchronization methods were two main reasons to take the different effects of superovulation among all the factors. The intercept effect was not significant.
Analysis on 6 kinds of influential factors for superovulation effect with One-way ANOVA Analysis and Nonparametric Test. The result showed : The CL number of the MM (more than middle level of body status ) and M(middle level of body status ) groups were more than other groups significantly (11.17?.29 and 6.22?.37).It showed that the main effect that defferent body status affected the superovulation was the CL number; The effect of superovulation of the 7th or 9th day in the estrous period was better among different injection time of FSH. It showed that the ovum vesicle growth wave of Boer Goats should begins with the 7~9th day in the estrous period. CL numbers were different significantly in the different method for synchronization (PO.01). PG 2ml (2 mL /once) was injected to donor, which affected the rate of fertilization and usable embryo number significantly. There were no difference between the bolt group and the injection group. The result showed that PG 2 mL (interval 12h, 1 mL /once) was injected, w
hich can produce perfect effect of superovulation and estrous synchronization; The rate of picking embryo (29.56%<63.58%) and the rate of usable embryo (75.30%<94.34%) in the injection group with LH were lower than no injection group, and the rate of no zygote (20.07%>1.37%)was higher than no injection group significantly. It showed that LH was injected to the donor, which increased the number of ovulation of premature follicular; The embryo number(6.29 ?1.57)and usable embryo number(5.10?1.56)in the 24 h estrus group (donor's estrous time was in the 24h after estrous synchronization)after synchronization were higher than the 36 h or 42 h estrous group, but the rate of no zygote (15.90%) was higher than other two groups.The difference for the effect of superovulation among three groups was no significant (P>0.05). It showed that the estrous time of donor couldn't affect the effect of superovulation significantly; Mating assistance with AI can reduce significantly the no zygote rate (PO.05), morever improve
significantly effect of
superovulation.
The rate of synchronization within 36 h of donor was 100% in the PG group. PG was injected 2 mL (interval 12 h , 1 mL /once )can make the rate of synchronization within 24 h reach to 80%; The correlated coefficient between the CL number and embryo number was 0.415(P<0.05), the correlated coefficient between the embryo number and usable embryo number was 0.928(P<0.01), we can predict the embryo number and usable embryo number through the CL number .
Four factors affecting the rate of recipient were discussed with Logistic Regression Model. The result showed that the method of transfer and the status of estrous affected the rate of recipient pregnancy signifi
采用适合性检验对5个超排指标进行正态性分析，结果表明：黄体数、可用胚数、采胚数均服从正态分布。而变性胚数和未受精卵数不服从正态分布，对其应采用非参数检验的方法；多变量方差分析对影响超排效果的6个因素的分析结果表明：FSH的注射时间和同期处理方法在综合效应模型中是产生超排效果差异的主要原因。各因素间的交互作用差异不显著。
采用单因素方差分析和非参数检验对影响超排效果的6个因素进行分析。结果表明：膘情因素四个水平中，中上、中等体况个体的黄体数为(11.17±3.29)枚、(6.22±2.37)枚，显著高于其它水平(P＜0.01)，而变性胚率和未受精卵率居较低水平。说明不同膘情对超排效果的影响主要体现在黄体数方面；三种FSH注射时间以第7d或第9d开始注射的超排效果好，说明波尔山羊的卵泡生长波的启动应该集中在第7～9d；不同的同期处理方法对黄体数量产生极显著差异(P＜0.01)，一次肌注PG2mL剂量过大，会严重影响供体的受精卵率和可用胚数。放栓组和PG注射组比较无显著差异(P＞0.05)。结果表明，PG分两次注射(1mL／次)的总体效果较好；注射LH后其采胚率(29.56％＜63.58％)、可用胚率(75.30％＜94.34％)低于不注射组，并且未受精卵率显著高于不注射组(20.07％＞1.37％)。说明LH的注射增加了不成熟卵泡的排卵数目；同期处理后24h发情的供体在采胚数(6.29±1.57)和可用胚数(5.10±1.56)方面高于36h、42h发情组，同时未受精率也较高(15.90％)，三个组对超排效果的影响差异不显著(P＞0.05)，说明供体发情出现的时间对超排效果的影响并不明显；人工+本交的交配方式可显著降低未受精卵率(P＜0.05)，提高超排效果。
PG的应用可使供体的36h同期发情率达到100％，各水平间以两次注射PG的效果为好，24h发情率达到80％；黄体数与采胚数的相关系数为0.415(P＜0.05)，采胚数和可用胚数的相关系数为0.928(P＜0.01)，在实际应用中可以通过黄体数判断其采胚数和可用胚数。
采用Logistic回归分析了影响受体妊娠率的4种因素，结果表明，移植方式和发情状况对妊娠结果的产生具有决定性作用(P＜0.01)，可通过建立的回归模型对受体妊娠情况进行预测。胚胎发育阶段的差异会极显著影响返情状况；自然发情的受体有较高的不返情率(69.44％)，显著高于诱导发情组(P＜0.05)；受体黄体数量的差异对返情率的影响不显著(P＞0.05)；双侧移植方式可显著提高受体的不返情率(P＜0.01)，达到了80％；桑椹胚的返情率最低，显著低于其它水平(P＜0.01)，达到31.58％。
综合以上研究结果，可以得出如下结论
门）FSH的注射时间和同期处理方法在综合效应模型中是产生超排效果差异的
主要原因。
② 采用如下方法可得到较好的超排效果：选择中上、中等体况的个体作为超
排个体，在第7 d或第9 d采用递减法注射FSH（16 nL），比例为S：5：3，每大2
次，间隔12小时；在超排第3 d上、下午各肌注lmLPG（间隔12 h）；选择同期处
理后 24 h发情的个体；采用本交十人工授精的交配方式；在发情羊配种后 6～7大，
采用于宫角冲胚法可得到较好的超排效果。
（3）采用如下方法可得到最好的妊娠效果：将采集到的桑棋胚移植到同期发情
（同步差为上12 h）的受体内。受体最好为自然发情：采用于宫角双侧移植的方法。
。＿。，、，。，、＿．．＿＿＿，，、、＿＿。、，，＿，＿、＿＿＿C
（4）用发情状况（XI）和移植方式（x；）建立的回归方程P＝＿
l＋e
可对受体妊娠率进行预测，预测准确率为79 3％。
In recent years, embryo transfer technology has made great contributions to increasing the amount of goats for breeding in a short period, but the success rate of embryo transfer was not stable in practice. Many people from native or abroad have carried out lots of study on improving the rate of success, but most of them discussed only about one factor affecting the rate of success. A comprehensive study on the influencial factors in the course of embryo transfer will help to improve the rate of success and enhance the stability of it. In this study, 30 Boer goats as donor; 89 LUBei white goats as recipients of estrus synchronization, analysis on varieties of factors affecting the superovulation, synchronization and the rate of pregnancy.
Analysis on 5 kinds of targets for superovulation by normal test showed: CL (corpus luteum) number, embryo number, usable embryo number were normal, while metamorphic embryo number and no zygote number were unnormal. So nonparametric test method should be adopted for the later; Multivariate Analysis on 6 kinds of influential factors for the superovulation effect showed: The injection time of FSH and estrous synchronization methods were two main reasons to take the different effects of superovulation among all the factors. The intercept effect was not significant.
Analysis on 6 kinds of influential factors for superovulation effect with One-way ANOVA Analysis and Nonparametric Test. The result showed : The CL number of the MM (more than middle level of body status ) and M(middle level of body status ) groups were more than other groups significantly (11.17?.29 and 6.22?.37).It showed that the main effect that defferent body status affected the superovulation was the CL number; The effect of superovulation of the 7th or 9th day in the estrous period was better among different injection time of FSH. It showed that the ovum vesicle growth wave of Boer Goats should begins with the 7~9th day in the estrous period. CL numbers were different significantly in the different method for synchronization (PO.01). PG 2ml (2 mL /once) was injected to donor, which affected the rate of fertilization and usable embryo number significantly. There were no difference between the bolt group and the injection group. The result showed that PG 2 mL (interval 12h, 1 mL /once) was injected, w
hich can produce perfect effect of superovulation and estrous synchronization; The rate of picking embryo (29.56%<63.58%) and the rate of usable embryo (75.30%<94.34%) in the injection group with LH were lower than no injection group, and the rate of no zygote (20.07%>1.37%)was higher than no injection group significantly. It showed that LH was injected to the donor, which increased the number of ovulation of premature follicular; The embryo number(6.29 ?1.57)and usable embryo number(5.10?1.56)in the 24 h estrus group (donor's estrous time was in the 24h after estrous synchronization)after synchronization were higher than the 36 h or 42 h estrous group, but the rate of no zygote (15.90%) was higher than other two groups.The difference for the effect of superovulation among three groups was no significant (P>0.05). It showed that the estrous time of donor couldn't affect the effect of superovulation significantly; Mating assistance with AI can reduce significantly the no zygote rate (PO.05), morever improve
significantly effect of
superovulation.
The rate of synchronization within 36 h of donor was 100% in the PG group. PG was injected 2 mL (interval 12 h , 1 mL /once )can make the rate of synchronization within 24 h reach to 80%; The correlated coefficient between the CL number and embryo number was 0.415(P<0.05), the correlated coefficient between the embryo number and usable embryo number was 0.928(P<0.01), we can predict the embryo number and usable embryo number through the CL number .
Four factors affecting the rate of recipient were discussed with Logistic Regression Model. The result showed that the method of transfer and the status of estrous affected the rate of recipient pregnancy signifi
引文
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