牛、羊胚胎移植技术研究与应用
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摘要
牛、羊胚胎移植技术中，供受体同期化程度不理想、超排效果不稳定以及相关指标不系统是制约其商业化应用的几个关键性问题。本研究以西北农林科技大学奶牛胚胎移植试验站为依托，与7家养殖公司协作进行，结果如下：
1.在荷斯坦奶牛应用“PG+P_4+FSH”的超排方案中，①不同去栓时间(分别在第7次和第8次注射FSH时去栓)卵巢平均黄体数(5.5±0.5个和6.0±1.3个)差异不显著(P＞0.05)；平均采胚数(4.0±0.0枚和7.5±3.3枚)差异不显著(P＞0.05)；平均可用胚数(3.0±0.0枚和2.8±0.9枚)差异不显著(P＞0.05)。②FSH(Folltropin-V，澳大利亚产)的剂量分别为400mg与440mg时，平均冲胚数分别为4.0±0.0枚和7.3±1.9枚，二者差异不显著(P＞0.05)。
2.在波尔山羊应用“PG+P_4+FSH”的超排方案中，①方案1与方案2(方案1比方案2去栓时间晚6 h)相比，两组平均黄体数(33.3±2.2个和23.5±1.6个)差异显著(P＜0.05)；但平均冲胚数(22.4±2.3枚和20.4±2.0枚)和平均可用胚数(20.9±2.0枚和18.8±2.1枚)差异不显著(P＞0.05)。②中科院动物所产FSH和澳大利亚产FSH(Folltropin-V)对波尔山羊超排的效果差异不显著(P＞0.05)；中科院产FSH(9mg与11mg)，澳大利亚产Folltropin-V(360mg与440mg)，不同剂量间的超排效果差异均不显著(P＞0.05)。③在陕西，8～11月份对波尔山羊进行超排(平均冲胚数分别为22.4±2.3枚、17.8±2.2枚和19.3±5.5枚；平均可用胚数分别为20.9±2.1枚、16.0±2.0枚和14.6±5.3枚)，其效果无显著差异(P＞0.05)。
3.以波尔山羊为供体、关中奶山羊为受体，当受体比供体提前12h，6h或同时去栓时的同期率无显著差异(73.8％、80.7％和73.1％，P＞0.05)。
4.受体羊(关中奶山羊)黄体体积的大小对胚胎移植妊娠率的影响差异不显著(黄体体积大、中、小对应的妊娠率分别为69.6％、56.0％和71.4％，P＞0.05)。
5.移植三胚、双胚与单胚的受体羊(关中奶山羊)间妊娠率差异不显著(78.8％、61.9％和61.8％，P＞0.05)。移双胚产双羔的百分率为32.4％(55/170)。
6.供体羊黄体为“白黄体”和正常黄体时，两者超排卵巢反应差异不显著(平均黄体数分别为16.2±1.1个和20.1±1.1个，P＞0.05)；平均冲胚数(4.3±2.4枚和17.1±1.3枚)和平均可用胚数(4.3±2.4枚和15.0±1.3枚)差异极显著(P＜0.01)；每只供体获羔羊数差异显著(1.7±0.9只和5.2±0.6只，P＜0.05)。“白黄体”组与正常黄体组胚胎移植妊娠率分别为为66.7％(8/12)和57.5％(207/360)，二者差异不显著(P＞0.05)。
The commercial application of embryo transfer (ET) in cattle and goats was restricted by the ordinary estrous synchronization, unstable superovulation and unsystematic related index. This research was supported by the Bovine Embryo Engineering Institute of NWSUAF and 7 Farms. The result was as follows:
1. In the bovine superovulation procedure of Holstein by using PG, P4 and FSH:(1) The number of corpus luteum (CL) was not significant different by different time removing the CIDR-B at the 7th and 8th FSH administrition(5.5±0.5 vs 6.0±1.3, P> 0.05). The number of total embryos (4.0 ±0. 0 vs 7.5 ±3.3) and transferable embryos (3.0±0.0 vs 2.8±0.9) were not significatly difference (P>0.05).(2) The Flushing total embryos(4.0±0.0vs 7.3 ±1.9) by using different Folltropin-V dose 400 and 440 mg respectively were not significantly different (P>0.05).
2. In the superovulation procedure of Boer goats by using PG, P4 and FSH, (1)In different procedures (Procedure 1 removes CIDR or PRID later 6h than procedure 2), the number of CL was significantly difference (33.3±2.2 vs 23.5±1.6, P<0.05) , the number of total embryos (22.4 ±2.3 vs 20.4 ±2.0) and transferable embryos (20.9 ± 2.0 vs 18.8 ±2.1) was not significantly (P>0.05). ㏕he results of superovulation hadn't distingushed difference using the different kinds of FSH (produced in Australia and Institute of Zoology, Chinese Academy of Sciences) and different dose(P>0.05). (3) The superovulatory results hadn't distinguished difference (total embryos was 22.4 ±2.3, 17.8±2.2 vs 19.3±5.5, transferable embryos was 20.9 ± 2.1, 16.0 ± 2.0 vs 14.6±5.3, P >0.05) from August to December in Shaanxi.
3. Boer goats were used as the donors and Guanzhong dairy goats as the recipients in the study. The estrous synchronization rates among the recipient's earlier 12h, 6h and oh were not significantly different (73.8%, 80.7% vs 73.1%, P>0.05).
4. In the studies of relationship between the volume of CL and pregnancy, the pregnancy rates between recipients with different size of CL were not significantly different (69.6%, 56.0% vs 71.4%, P>0.05).
5. The pregnancy rate of the recipients transfered 1, 2, 3 embryos was not significantly different (78.8%, 61.9% vs 61.8%, P>0.05). The recipient's ratio of producing twin kids which transfered 2 embryos was 32.4% (55/170).
6. The ovarian responses between the "white" and normal CL goats were not significantly different (the numbers of CL are 16.2±1.1 vs 20.1 ±1.1 respectively, P> 0.05). Total embryos (4.3 ±2.4 vs 17.1 ±1.5) and transferable embryos (4.3 ±2.4 vs 15.0 ±1.3) were significantly different (P<0.01). The kids of every doner were significantly different (1.7±0.9 vs 5.2 ± 0.6, P<0.05). The pregnancy rates of recipients were not affected (66.7% vs 57.5%, P>0.05).
1.在荷斯坦奶牛应用“PG+P_4+FSH”的超排方案中，①不同去栓时间(分别在第7次和第8次注射FSH时去栓)卵巢平均黄体数(5.5±0.5个和6.0±1.3个)差异不显著(P＞0.05)；平均采胚数(4.0±0.0枚和7.5±3.3枚)差异不显著(P＞0.05)；平均可用胚数(3.0±0.0枚和2.8±0.9枚)差异不显著(P＞0.05)。②FSH(Folltropin-V，澳大利亚产)的剂量分别为400mg与440mg时，平均冲胚数分别为4.0±0.0枚和7.3±1.9枚，二者差异不显著(P＞0.05)。
2.在波尔山羊应用“PG+P_4+FSH”的超排方案中，①方案1与方案2(方案1比方案2去栓时间晚6 h)相比，两组平均黄体数(33.3±2.2个和23.5±1.6个)差异显著(P＜0.05)；但平均冲胚数(22.4±2.3枚和20.4±2.0枚)和平均可用胚数(20.9±2.0枚和18.8±2.1枚)差异不显著(P＞0.05)。②中科院动物所产FSH和澳大利亚产FSH(Folltropin-V)对波尔山羊超排的效果差异不显著(P＞0.05)；中科院产FSH(9mg与11mg)，澳大利亚产Folltropin-V(360mg与440mg)，不同剂量间的超排效果差异均不显著(P＞0.05)。③在陕西，8～11月份对波尔山羊进行超排(平均冲胚数分别为22.4±2.3枚、17.8±2.2枚和19.3±5.5枚；平均可用胚数分别为20.9±2.1枚、16.0±2.0枚和14.6±5.3枚)，其效果无显著差异(P＞0.05)。
3.以波尔山羊为供体、关中奶山羊为受体，当受体比供体提前12h，6h或同时去栓时的同期率无显著差异(73.8％、80.7％和73.1％，P＞0.05)。
4.受体羊(关中奶山羊)黄体体积的大小对胚胎移植妊娠率的影响差异不显著(黄体体积大、中、小对应的妊娠率分别为69.6％、56.0％和71.4％，P＞0.05)。
5.移植三胚、双胚与单胚的受体羊(关中奶山羊)间妊娠率差异不显著(78.8％、61.9％和61.8％，P＞0.05)。移双胚产双羔的百分率为32.4％(55/170)。
6.供体羊黄体为“白黄体”和正常黄体时，两者超排卵巢反应差异不显著(平均黄体数分别为16.2±1.1个和20.1±1.1个，P＞0.05)；平均冲胚数(4.3±2.4枚和17.1±1.3枚)和平均可用胚数(4.3±2.4枚和15.0±1.3枚)差异极显著(P＜0.01)；每只供体获羔羊数差异显著(1.7±0.9只和5.2±0.6只，P＜0.05)。“白黄体”组与正常黄体组胚胎移植妊娠率分别为为66.7％(8/12)和57.5％(207/360)，二者差异不显著(P＞0.05)。
The commercial application of embryo transfer (ET) in cattle and goats was restricted by the ordinary estrous synchronization, unstable superovulation and unsystematic related index. This research was supported by the Bovine Embryo Engineering Institute of NWSUAF and 7 Farms. The result was as follows:
1. In the bovine superovulation procedure of Holstein by using PG, P4 and FSH:(1) The number of corpus luteum (CL) was not significant different by different time removing the CIDR-B at the 7th and 8th FSH administrition(5.5±0.5 vs 6.0±1.3, P> 0.05). The number of total embryos (4.0 ±0. 0 vs 7.5 ±3.3) and transferable embryos (3.0±0.0 vs 2.8±0.9) were not significatly difference (P>0.05).(2) The Flushing total embryos(4.0±0.0vs 7.3 ±1.9) by using different Folltropin-V dose 400 and 440 mg respectively were not significantly different (P>0.05).
2. In the superovulation procedure of Boer goats by using PG, P4 and FSH, (1)In different procedures (Procedure 1 removes CIDR or PRID later 6h than procedure 2), the number of CL was significantly difference (33.3±2.2 vs 23.5±1.6, P<0.05) , the number of total embryos (22.4 ±2.3 vs 20.4 ±2.0) and transferable embryos (20.9 ± 2.0 vs 18.8 ±2.1) was not significantly (P>0.05). ㏕he results of superovulation hadn't distingushed difference using the different kinds of FSH (produced in Australia and Institute of Zoology, Chinese Academy of Sciences) and different dose(P>0.05). (3) The superovulatory results hadn't distinguished difference (total embryos was 22.4 ±2.3, 17.8±2.2 vs 19.3±5.5, transferable embryos was 20.9 ± 2.1, 16.0 ± 2.0 vs 14.6±5.3, P >0.05) from August to December in Shaanxi.
3. Boer goats were used as the donors and Guanzhong dairy goats as the recipients in the study. The estrous synchronization rates among the recipient's earlier 12h, 6h and oh were not significantly different (73.8%, 80.7% vs 73.1%, P>0.05).
4. In the studies of relationship between the volume of CL and pregnancy, the pregnancy rates between recipients with different size of CL were not significantly different (69.6%, 56.0% vs 71.4%, P>0.05).
5. The pregnancy rate of the recipients transfered 1, 2, 3 embryos was not significantly different (78.8%, 61.9% vs 61.8%, P>0.05). The recipient's ratio of producing twin kids which transfered 2 embryos was 32.4% (55/170).
6. The ovarian responses between the "white" and normal CL goats were not significantly different (the numbers of CL are 16.2±1.1 vs 20.1 ±1.1 respectively, P> 0.05). Total embryos (4.3 ±2.4 vs 17.1 ±1.5) and transferable embryos (4.3 ±2.4 vs 15.0 ±1.3) were significantly different (P<0.01). The kids of every doner were significantly different (1.7±0.9 vs 5.2 ± 0.6, P<0.05). The pregnancy rates of recipients were not affected (66.7% vs 57.5%, P>0.05).
引文
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