提高牛、羊胚胎移植效率的因素分析
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摘要
本实验目的是通过对影响牛羊胚胎移植妊娠率和影响绵山羊超排效果以及影响绵羊同期发情效果的因素分析，建立一套比较完整的技术体系。本实验对800头母牛进行同期发情处理，胚胎移植600头；对绵山羊超数排卵和新鲜胚胎移植实验，共移植受体绵羊466只，山羊913只；在绵羊冷冻胚胎移植实验中，同期发情处理受体705只，移植受体460只。结果表明：西杂牛的移植利用率和移植妊娠率高于土种杂交牛和黑白花，但各组间差异不显著(P＞0.05)：2～6周岁的受体牛移植利用率和移植妊娠率高于≥7岁的受体牛和＜2岁的受体牛，但各组间差异不显著(P＞0.05)；经产1～3胎牛高于育成牛的利用率和妊娠率，但各组间差异不显著(P＞0.05)；并且利用率显著高于≥4胎的受体牛利用率，组间差异显著(P＜0.05)；体况膘情上等和中等的受体牛移植利用率和妊娠率差异不显著(P＞0.05)；同期发情放置阴道栓时卵巢处于黄体期的利用率显著高于处于卵泡期的利用率，两者之间差异极显著(P＜0.01)；移植后是否再次放置阴道栓对移植妊娠率的影响不大(P＞0.05)；双胚的移植妊娠率高于单胚，但差异不显著(P＞0.05)；早期囊胚的移植妊娠率最高，其次为桑椹胚，最低为囊胚，但各组间差异不显著(P＞0.05)；左侧子宫角的妊娠妊娠率显著高于右侧(P＜0.05)。FSH(中科院产)和Ovagen及新FSH对道塞特、萨福克、杜伯等的超排效果，平均可用胚数之间只有道塞特表现出FSH(中科院产)显著高于Ovagen(P＜0.05)；澳FSH、加FSH、新FSH对波尔山羊的超排结果表明，加FSH的可用胚数高于新FSH，差异不显著(P＞0.05)，并且显著高于澳FSH，差异极显著(P＜0.01)；在重复超排实验中，波尔山羊两次都获得8枚以上的可用胚数，绵羊两次超排可用胚数间差异不显著(P＞0.05)；山羊受体卵巢有无卵泡对移植妊娠率差异不显著(P＞0.05)；绵山羊受体单侧3个黄体的移植妊娠率高于2个黄体的和1个黄体，2个黄体高于1个黄体的移植妊娠率，但各组间差异不显著(P＞0.05)；1M+的移植妊娠率显著高于1M的妊娠率(P＜0.05)：1M+1B与2M、2B间的移植妊娠率差异不显著(P＞0.05)；腹腔镜与手术法间移植妊娠率无显著性差异(P＞0.05)；用CIDR和Progestagen Sponge(孕酮海绵栓)处理绵羊同期发情，发情率无显著性差异(P＞0.05)。
Factors on the pregnancy rate of bovine or ovine embryos transferred, superovulation of sheep and goats, and estrus synchronization of sheep to establish a highly efficient ET system. In the experiment, of the 800 cows selected at random for estrus synchronization, 600 served as ET recipients. The sheep and goats fresh embryos collected following superovulation were transferred into recipient sheep
(n=466) and goats (n=913) . And 705 sheep were selected for estrus synchronization fol lowed by transfer of frozen-thawed embryos into 460 recipients. The results of the experiment indicated: The rate of available recipients and pregnancy rate in Simental cross-bred (multiparity cows (1~3 parity) /cows of 2 ~ 6 years) was higher than the local cattle cross-bred and Holsteins (nulliparity cows/the recipient cows (> 7 years or < 2 years)) , but they showed no significant difference (P>.05) between groups. The rate of available recipients of l~3-parity cows was higher(P<0.05) than that of >-parity cows. No significant difference (P>.05 ) in the rate of available recipients and pregnancy rate was observed in the cows of good condition and medium. The rate of available recipients treated for estrus synchronization with CIDR when ovum was in luteal stage was higher (PO.01) than that in follicular stage. Whether CIDR was inserted into recipient vagina or not after ET had no great effect on ET pregnanc
y rate (P>0.05 ). The pregnancy rate of double embryo transferred was higher (P>0.05 ) than that of single one. Early blastocyst had the highest pregnancy rate in ET, the second was morula, and the last was blastocyst; no significance (P>0.05) was observed between groups. The pregnancy rate of embryo transferred into left-side uterus horn was higher (PO.05) than that of right-side one.The mean number of viable embryos collected from Dorset superovulated by highly purified FSH made in CAS (Chinese Academy of Science ) was obviously higher (P<0.05 ) than that from Ovagen. Furthermore the mean number of viable embryos collected form Boer goats superovulated by highly purified FSH made in Canada was significantly higher than in Australia (PO.01) and in Singapore (P>0.05) .After repeated superovulation, every Boer goat produced viable embryos (n > 8 ) in the first or second batch, and there was no significant difference (P>0.05) in the two bacthes of superovulation of sheep producing viable embryos. Preg
nancy rate of recipient goats with or without follicle was similar (P>0.05). Pregnancy rate of recipient sheep and goats with 3 CLs on single side was higher (P>0.05) than with 1 (the lowest) or 2 CLs. Pregnancy rate of 1M+ transferred was higher (P<0.05) than that of 1M. Pregnancy rate of 1M+1B, 2M and 2B transferred was similar (P>0.05) .No significant difference (P>0.05 ) was observed in the pregnancy rate of embryo transferred surgically or by laparoscope and the estrus rate of sheep treated with CIDR or Progestagen Sponge did not differ (P>0.05 ) .
Factors on the pregnancy rate of bovine or ovine embryos transferred, superovulation of sheep and goats, and estrus synchronization of sheep to establish a highly efficient ET system. In the experiment, of the 800 cows selected at random for estrus synchronization, 600 served as ET recipients. The sheep and goats fresh embryos collected following superovulation were transferred into recipient sheep
(n=466) and goats (n=913) . And 705 sheep were selected for estrus synchronization fol lowed by transfer of frozen-thawed embryos into 460 recipients. The results of the experiment indicated: The rate of available recipients and pregnancy rate in Simental cross-bred (multiparity cows (1~3 parity) /cows of 2 ~ 6 years) was higher than the local cattle cross-bred and Holsteins (nulliparity cows/the recipient cows (> 7 years or < 2 years)) , but they showed no significant difference (P>.05) between groups. The rate of available recipients of l~3-parity cows was higher(P<0.05) than that of >-parity cows. No significant difference (P>.05 ) in the rate of available recipients and pregnancy rate was observed in the cows of good condition and medium. The rate of available recipients treated for estrus synchronization with CIDR when ovum was in luteal stage was higher (PO.01) than that in follicular stage. Whether CIDR was inserted into recipient vagina or not after ET had no great effect on ET pregnanc
y rate (P>0.05 ). The pregnancy rate of double embryo transferred was higher (P>0.05 ) than that of single one. Early blastocyst had the highest pregnancy rate in ET, the second was morula, and the last was blastocyst; no significance (P>0.05) was observed between groups. The pregnancy rate of embryo transferred into left-side uterus horn was higher (PO.05) than that of right-side one.The mean number of viable embryos collected from Dorset superovulated by highly purified FSH made in CAS (Chinese Academy of Science ) was obviously higher (P<0.05 ) than that from Ovagen. Furthermore the mean number of viable embryos collected form Boer goats superovulated by highly purified FSH made in Canada was significantly higher than in Australia (PO.01) and in Singapore (P>0.05) .After repeated superovulation, every Boer goat produced viable embryos (n > 8 ) in the first or second batch, and there was no significant difference (P>0.05) in the two bacthes of superovulation of sheep producing viable embryos. Preg
nancy rate of recipient goats with or without follicle was similar (P>0.05). Pregnancy rate of recipient sheep and goats with 3 CLs on single side was higher (P>0.05) than with 1 (the lowest) or 2 CLs. Pregnancy rate of 1M+ transferred was higher (P<0.05) than that of 1M. Pregnancy rate of 1M+1B, 2M and 2B transferred was similar (P>0.05) .No significant difference (P>0.05 ) was observed in the pregnancy rate of embryo transferred surgically or by laparoscope and the estrus rate of sheep treated with CIDR or Progestagen Sponge did not differ (P>0.05 ) .
引文
Adams G P, 1994. Control of ovarian follicular wave dynamics in cattle: implications for synchronization & superstimulation. Therigenology, 41:19-24
Adams GP Control of ovarian follicular wave dynamics in cattle: implications for synchroization and superstimulation. Theriogenology 1994; 41: 19-24。
Adams GP et al. Superstimulatory response of ovarian follicles of wave 1 versus wave 2 in heifers. Theriogenology 1994:42:1103-1113
AGP et al. Selection of the dominant follicle on regression of its subordinates in heifers Can J Anim Sci 1993;73:267-275
Arije B.G et al. Prediction of age and weight at puberty in beef heifers. J Anim. Sci 1974;38:803-810.
Armstrong D T, 1993, Recent advances in superovulation ofcattle. Theriogenology, 39:7-24。
Armstrong D T, Pfitzner A P, Warnes O M, et al. Superovulation treatment and embryo transfer in Angora goats[J]. Journal of Reproduciton and Fertility, 1983, 67(2):403-410.
Armstrong DT, Evans G. Factors influencing success of embryo transfer in sheep and goats Theriogenology Teriogenology 1983;19;31-42.
Barros CM et al. Synchronization of ovulation in beef cows (Bos indicus) using GnRH, PGF2 a and estrdiol benzoate Theriogenology 2000, 53:1121-1134.
Beal WE et al. Variation in Pregnancy rate following synchronization of estrus with melengestrol acetate and prostaglandin F2 a. J Anim Sci 1988;66:599-602.
Biggers, B.G et al. Effect of heat stress on early embryonic development in the beef cow J.Anim. Sci. 1987;64:1512-1518
Bo G.A et al. Follicular wave emergence after treatment with estradiol benzoate and CIDR-B Vaginal devices in beef cattle. 13th Int Congr Anim Reprod Syney, Australia.
Bo GA et al. Effect of progestogen plus E-17 B treatment on superovulatory response in beef cattle. Theriogenology 1996;45:897-910.
Boland. Comparison of lambing out come in ewes after synchronization of estrus by progestagen or prostaglandin treatment[J].J. Agric. Sci. cama, 1978 (91) :756-766.
Boland. The effects of treatment by progtagandin analogue orprogestagen on ovulation and fertilization in cyclic ewes[J]. J.Agric. Sci. 1987 (91):727-730.
Bondioli K M et al. Theriogenology., 1989; 31:95-104
Booth WD et al. Plasma oestrogen and progesterone in relation to superovulation and egg recovery in the cow. Vet Rec 1975;97;366-369
Brebion P, Baril G, Cognie Y, Vallet JC. Embryo transfer in sheep and goat. Ann Zoot 1992;41:331-339.
Brebion P, Cognie Y. Increased superovulation in the ewe following 14 days of GnRH agonist pre-treatment.In:Proc 5th Mtg Ass Eur Trans Emb. (AETE) 1989;106abstr.
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Broadbent P J, Stewart M, Dolman D F, 1991, Recipient managem ent and embryo transfer. Theriogemology, 35:115-126
Bulter W R. et al. Imterrelationships between energy balance and postpartum reproductive function in dairy cattle. J.Dairy Sci 1989;72:767-783
Calder M, Rajamahendran R. follicular growth, ovulation and embryo recovery in dairy cows given FSH at the beginning or middle of the estrus cycle. Therioenology 1992;38:1163-1174
Christie W.B et al. Embryo survival in heifers after transfer of an gee to the uterine horn contralateral to the corpus luteum and the effect of treatment with progesterone or HCG on pregnancy rats. J.Reprod. Fert. 1979;56:701-706
Cognie Y, Benoit F, Poulin N, et al. Effect of follicle size and of the FecB Booroola gene on oocyte fumction in sheep. J Reprod Fertil 1998;112:379-386.
Colazo EE et al. Fixed-time artificial insemination in beef cattle using CIDR-B devices, progesterone and estriodiol benzoate. Theriogenology 1995;51:404 abstr
Coleman, DA et al. Estrus synchronization and establishment of pregnancy in bovine embryo transfer recipients J Dairy Sci. 1987;70:858-866
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Adams GP Control of ovarian follicular wave dynamics in cattle: implications for synchroization and superstimulation. Theriogenology 1994; 41: 19-24。
Adams GP et al. Superstimulatory response of ovarian follicles of wave 1 versus wave 2 in heifers. Theriogenology 1994:42:1103-1113
AGP et al. Selection of the dominant follicle on regression of its subordinates in heifers Can J Anim Sci 1993;73:267-275
Arije B.G et al. Prediction of age and weight at puberty in beef heifers. J Anim. Sci 1974;38:803-810.
Armstrong D T, 1993, Recent advances in superovulation ofcattle. Theriogenology, 39:7-24。
Armstrong D T, Pfitzner A P, Warnes O M, et al. Superovulation treatment and embryo transfer in Angora goats[J]. Journal of Reproduciton and Fertility, 1983, 67(2):403-410.
Armstrong DT, Evans G. Factors influencing success of embryo transfer in sheep and goats Theriogenology Teriogenology 1983;19;31-42.
Barros CM et al. Synchronization of ovulation in beef cows (Bos indicus) using GnRH, PGF2 a and estrdiol benzoate Theriogenology 2000, 53:1121-1134.
Beal WE et al. Variation in Pregnancy rate following synchronization of estrus with melengestrol acetate and prostaglandin F2 a. J Anim Sci 1988;66:599-602.
Biggers, B.G et al. Effect of heat stress on early embryonic development in the beef cow J.Anim. Sci. 1987;64:1512-1518
Bo G.A et al. Follicular wave emergence after treatment with estradiol benzoate and CIDR-B Vaginal devices in beef cattle. 13th Int Congr Anim Reprod Syney, Australia.
Bo GA et al. Effect of progestogen plus E-17 B treatment on superovulatory response in beef cattle. Theriogenology 1996;45:897-910.
Boland. Comparison of lambing out come in ewes after synchronization of estrus by progestagen or prostaglandin treatment[J].J. Agric. Sci. cama, 1978 (91) :756-766.
Boland. The effects of treatment by progtagandin analogue orprogestagen on ovulation and fertilization in cyclic ewes[J]. J.Agric. Sci. 1987 (91):727-730.
Bondioli K M et al. Theriogenology., 1989; 31:95-104
Booth WD et al. Plasma oestrogen and progesterone in relation to superovulation and egg recovery in the cow. Vet Rec 1975;97;366-369
Brebion P, Baril G, Cognie Y, Vallet JC. Embryo transfer in sheep and goat. Ann Zoot 1992;41:331-339.
Brebion P, Cognie Y. Increased superovulation in the ewe following 14 days of GnRH agonist pre-treatment.In:Proc 5th Mtg Ass Eur Trans Emb. (AETE) 1989;106abstr.
Bredbacka P et al. Pcr-sexing of bovine embryos:a simplified protocol. Theriogenology 1995;44:167-176
Broadbent P J, Stewart M, Dolman D F, 1991, Recipient managem ent and embryo transfer. Theriogemology, 35:115-126
Bulter W R. et al. Imterrelationships between energy balance and postpartum reproductive function in dairy cattle. J.Dairy Sci 1989;72:767-783
Calder M, Rajamahendran R. follicular growth, ovulation and embryo recovery in dairy cows given FSH at the beginning or middle of the estrus cycle. Therioenology 1992;38:1163-1174
Christie W.B et al. Embryo survival in heifers after transfer of an gee to the uterine horn contralateral to the corpus luteum and the effect of treatment with progesterone or HCG on pregnancy rats. J.Reprod. Fert. 1979;56:701-706
Cognie Y, Benoit F, Poulin N, et al. Effect of follicle size and of the FecB Booroola gene on oocyte fumction in sheep. J Reprod Fertil 1998;112:379-386.
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