肉羊同期发情和人工授精技术的研究与示范
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摘要
本文所提肉羊是指肉用绵羊品种,以下如无特别说明,所提肉羊均为肉用绵羊。
为了提高引进优良肉用种公羊的利用率,迅速扩大肉羊的种群和繁育数量,本试验对肉羊同期发情和人工授精的配套繁殖技术进行了系统研究。研究内容包括母羊同期发情技术、公羊精液低温保存方法、精液冷冻稀释液筛选、种公羊的饲养模式及人工授精方法。通过以上内容的研究,建立了从同期发情到人工授精的肉羊高效繁殖配套技术体系,并对肉羊产业化生产进行推广。母羊同期发情方法进行试验和研究,并对传统的阴道栓法的各个环节进行了认真试验和细致研究,对阴道栓法的某些技术环节进行了改进,通过对母羊同期发情技术的改进,使用阴道栓法处理母羊同期发情,具有发情率高,易于操作,成本低,效果好,受胎率高,适于产业化推广的特点,是母羊同期发情处理的首选方法;不同饲养管理对种公羊精液活力存在影响,对肉羊鲜精低温保存和精液冷冻效果影响显著,通过实验,放牧结合补饲精料的饲养方式有利于提高种公羊精液精子的活力和精液的品质;通过实验筛选出了适宜生产中应用的精液低温保存稀释液配方和保存方法,即葡萄糖-果糖-庶糖-蛋黄稀释液;不同冷冻稀释液的冷冻效果不同,本实验对五种冷冻稀释液进行实验和分析,筛选出了稀释液二适合对肉羊精液进行稀释,用于制作冷冻精液;对传统输精方法进行改进,用自制的羊阴道观察器(已申请了国家专利)判断母羊是否发情和确定最佳输精时间,并用特制的输精枪,显著提高了羊人工输精的受胎率。
In order to make use of foreign introduced ram effectively, and to enlarge the breeding population, we carried out systemic studies on cmprehensive reproduction techniques of ram production including ewe synchronized estrus technique, ram breeding and management models, semen low-temperature preservation methods, semen cryopreservation diluents and artificial insemination methods.
Three ewe synchronized estrus methods were studied to investigate their validity. Results showed that fertility efficiencies of using a vaginal plug and of sex hormone 1 are significantly higher than that of sex hormone 2 (P<0.01), while there is no significant difference between that of vaginal plug and of sex hormone 1 methods. Considering the simpleness in operation and low cost of the vaginal plug method , it is the best choice in ewe synchronized estrus treatment.
Four breeding and management models were investigated to research their effects on ram semen quality. Results showed that breeding and management model plays great effect on ram semen low-temperature preservation quality. The model of feed supplementating plus grazing can improve ram semen quality.
Three low-temperature preservation diluents were studied to compare their efficiencies. Results showed that the the sperm viability of using milk diluent is significant lower than that of using diluent A and C (P<0.01), while there is no significant difference between t diluent A and C. According to this experiment, ram semen can be preserved for 167 hours at low temperature in diluent A. The sperm viability after 77 hours is 70%. The living time of ram sperm at room temperature is about 72h, thus low-temperature preservation can greatly prolong the living time of ram sperm.
Five semen cryopreservation diluents were screened in this experiment. Results showed that there are significant difference of these five diluents on the effect of frozen sperm (P<0.05). the post-thawed sperm viability of diluent 2, 3 and 4 is significantly higher than that of diluent1nd 5(P<0.05).
In order to improve the conception rate of frozen semen, traditional insemination method was reformed. Results showed that the average conception rate of using the reform insemination method is 62.7%, which is significantly higher than that of the traditional method, which is only 32.3% P<0.01).
为了提高引进优良肉用种公羊的利用率,迅速扩大肉羊的种群和繁育数量,本试验对肉羊同期发情和人工授精的配套繁殖技术进行了系统研究。研究内容包括母羊同期发情技术、公羊精液低温保存方法、精液冷冻稀释液筛选、种公羊的饲养模式及人工授精方法。通过以上内容的研究,建立了从同期发情到人工授精的肉羊高效繁殖配套技术体系,并对肉羊产业化生产进行推广。母羊同期发情方法进行试验和研究,并对传统的阴道栓法的各个环节进行了认真试验和细致研究,对阴道栓法的某些技术环节进行了改进,通过对母羊同期发情技术的改进,使用阴道栓法处理母羊同期发情,具有发情率高,易于操作,成本低,效果好,受胎率高,适于产业化推广的特点,是母羊同期发情处理的首选方法;不同饲养管理对种公羊精液活力存在影响,对肉羊鲜精低温保存和精液冷冻效果影响显著,通过实验,放牧结合补饲精料的饲养方式有利于提高种公羊精液精子的活力和精液的品质;通过实验筛选出了适宜生产中应用的精液低温保存稀释液配方和保存方法,即葡萄糖-果糖-庶糖-蛋黄稀释液;不同冷冻稀释液的冷冻效果不同,本实验对五种冷冻稀释液进行实验和分析,筛选出了稀释液二适合对肉羊精液进行稀释,用于制作冷冻精液;对传统输精方法进行改进,用自制的羊阴道观察器(已申请了国家专利)判断母羊是否发情和确定最佳输精时间,并用特制的输精枪,显著提高了羊人工输精的受胎率。
In order to make use of foreign introduced ram effectively, and to enlarge the breeding population, we carried out systemic studies on cmprehensive reproduction techniques of ram production including ewe synchronized estrus technique, ram breeding and management models, semen low-temperature preservation methods, semen cryopreservation diluents and artificial insemination methods.
Three ewe synchronized estrus methods were studied to investigate their validity. Results showed that fertility efficiencies of using a vaginal plug and of sex hormone 1 are significantly higher than that of sex hormone 2 (P<0.01), while there is no significant difference between that of vaginal plug and of sex hormone 1 methods. Considering the simpleness in operation and low cost of the vaginal plug method , it is the best choice in ewe synchronized estrus treatment.
Four breeding and management models were investigated to research their effects on ram semen quality. Results showed that breeding and management model plays great effect on ram semen low-temperature preservation quality. The model of feed supplementating plus grazing can improve ram semen quality.
Three low-temperature preservation diluents were studied to compare their efficiencies. Results showed that the the sperm viability of using milk diluent is significant lower than that of using diluent A and C (P<0.01), while there is no significant difference between t diluent A and C. According to this experiment, ram semen can be preserved for 167 hours at low temperature in diluent A. The sperm viability after 77 hours is 70%. The living time of ram sperm at room temperature is about 72h, thus low-temperature preservation can greatly prolong the living time of ram sperm.
Five semen cryopreservation diluents were screened in this experiment. Results showed that there are significant difference of these five diluents on the effect of frozen sperm (P<0.05). the post-thawed sperm viability of diluent 2, 3 and 4 is significantly higher than that of diluent1nd 5(P<0.05).
In order to improve the conception rate of frozen semen, traditional insemination method was reformed. Results showed that the average conception rate of using the reform insemination method is 62.7%, which is significantly higher than that of the traditional method, which is only 32.3% P<0.01).
引文
[1] 张嘉保. 21 世纪我国家畜繁育技术的发展趋势与展望.黑龙江动物繁殖,1999,4
[2] 曾培坚等.应用 MOET 繁育中国美利奴优质细毛羊.中国畜牧杂志,1997,3
[3] 杨永林等.不同超排方法对中国美利奴羊超排效果试验.中国养羊,1997,4
[4] 杨永林等.利用同期发情技术提高中国美利奴羊双胎率的试验.黑龙江动物繁殖,1998,1
[5] 倪健宏等.不同药物在绵羊同期发情中的应用试验.黑龙江动物繁殖,1997,2
[6] 杨永林等.中国美利奴羊非繁殖季节同期发情、冻胚移植试验.中国草食动物,1996,2
[7] 陈学进.最有前途的家畜育种方法.草食家畜 1996
[8] 刘志芬等.浅谈山羊冷配受胎率.黑龙江动物繁殖,2003,1
[9] 刘玉堂.精液冷冻保存的影响因素分析.黑龙江动物繁殖,2003,2
[10] 张国昌.羊人工授精站操作程序.黑龙江动物繁殖,2003,3
[11] 幸奠权.提高母羊繁殖力综合技术.黑龙江动物繁殖,2003,4
[12] 刘俊方 艾群等.羊的工人授精.黑龙江动物繁殖,2004,1
[13] 鞠连民.提高冷冻精液配种效果的体会.黑龙江动物繁殖,2004,4
[14] 刘孝德,姜勋平.山羊精液常温稀释液和最佳稀释液倍数研究.中国草食动物,2003,5
[15] 罗海玲,贾志海.维生素 E 对绵羊鲜精及冻精精液品质的影响.中国草食动物,2004,5
[16] 吴世林.用维生素 B 注射液作解冻液能提高情期受胎率.畜牧兽医杂志,1989 年第 1
[17] 吴永孝,郝正里.中式解冻液解冻牛精液的受胎试验.黄牛杂志,1995,20-21
[18].吴永孝等.牛冻精中药解冻液首次开发成功.中国农业科学,1992, 25(2)
[19] 新疆农垦绵羊冻精科研协作组.提高绵羊冷冻精液受胎率的研究.绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[20] 新疆自治区绵羊冻精技术研究协作组.绵羊冷冻精液技术试验研究报告(1975-1980).绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[21] 张春兰等.小尾寒羊精子内磷脂在冷冻一解冻前后含量的变化.黑龙江畜牧科技,1999,3
[22] 张坚中,陈元明等.全国绵羊冷冻精液技术中间试验北京片预备试验报告.绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[23] 张连峰等.冷冻前后牛羊精子表面三种凝集素受体的定量研究.中国农业科学,1994,27(1)
[24] 张文举,杨军祥等.谷肤甘肤提高牛冻精活力及受胎率的研究.甘肃畜牧兽医,1997 年 2
[25] 张扬.用 VB 作颗粒冻精解冻液提高母牛冻配受胎率的研究.草食家畜,1997,2
[26] 赵宗胜,代江生等.维生素 B 作鲜精稀释液提高绵羊产羔性能试验.草与畜杂志,1998,2
[27] 朱裕鼎,张坚中等.提高绵羊冷冻精液质量和受胎率的研究.绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[28] 朱云彬.热滞蛋白在低温冷冻保存卵母细胞和胚胎中的作用.国外畜牧科技第 22 卷,2
[29] 安民家.畜精液的液态保存.家畜繁殖技术进展,1979,39
[30] Berger T,ally M.In vitro pertility evaluation ofcryoperserved ram semen and its corr Probnis E.Z., et al. Evaluation of relation fertility of cryopreserved goat sperm fertility of cryopreserved goat sperm. Theriogenology 41:711-717,1994
[31] Choudhry T. M.,Berger T.& D.elation with relative in vivo fertility.Theriogenology 44:1195-1200 1995
[32] Codde J,M&Berger T..In vivo fertilip, of rants in relation to sperm-zonapellucida binding and sperm-zona pellucida penetration of ovine oodles,Theriogenology 44:901-906, 1995
[33] Correa J.R&Zavos P.M.The h)poosrnotic swelling test: its employment as anassay to evaluate the functional integrity of the.f-ozen-thecnsed bovine spermmembrane Theriogenology 42:351-360,1994
[34] Correa J.R.&Zzvos P.M., Frozen-thawed bovine spermatozoa diluted,slowor rapid dilution method. measurements on occurrence of osmotic shock andsperm viability, Theriogenology 44:963-971, 1995
[35] Cross N.L..&Watson S.K., assessing acrosomal status of bovine sperm usingfluoresceinated lectins, Theriogenolo_y 42:89-98, 1994
[36] Deleeuw F.E. et al. Efects of various cryoprotective agents and membranestabilizing compounds on bull sperm membrane integrity after cooling andfreezing. Cryobiology, 1993, 30: 32-44
[37] Lenz R.W.. Ball G.D. Et Al. Chodroitin sulfate facilitates an acrosome reactionin bovine spermatozoa as evidenced by light microscopyelectron microscopy and vitro fertilization, Biol. Reprod. 1983; 28:683-690
[38] Mahaclevan M. et al. Efect of protective media and dilution methods。。preservation ofhuman spermatozoa. Andrologia, 1983, 15: 355-368
[39] Maxwell W.M.C., Landers A.J.&Evans G., Survival and fertility of ramspermatozoa frozen in pellets, straws and minitubes
[40] Molinia F. C., Evans G.&Maxwell, W.M.C., Incorporation of penetrating cryoprotectants in diluents for spermatozoa Theriogenology42:849-858,1994
[41] Maxwell W.M.C., Szell A., Hunton Jr,. Artificial breeding: embryotransfer and cloning. In: Oldham c.nt, martin gb, purvis iw (eds) reproduction ofmerino sheep-concepts and consequences. Ch. 17. Perth, School Of Agriculture(Animal Science), The University Of Western Australia, 1990; 217-237
[42] Nagy Sz,Hazas G.. Bali Papp A. et al. Evaluation of sperm tail menbraneintegrity 妙 the microscope Theriogenology 52:1153-1159,1999Polge C et al. Revival of spermatozoa after vitrification and dehydration at low temperature. Nature, 1949, 164:666
[43] Polge C et al. Revival of spermatozoa after vitrification and dehydration at low temperature. Nature, 1949, 164:666
[44] Sinha M.P.. Sinha A.K.&Singh B.K.. Effect of met 勿 Ixanthines of motility and fertility o f /cozen-thawed goat semen. Theriogenology 44:907-914. 1995
[45] Sutiyotin P.. Thwaites C.J.. Sanehez-Partida L.G.&B.P. Setchell. Evaluation of a modified sperm penetration test in ran? semen.Theriogenology 44 29-40 1995
[46] Galcarcel A., De Las Heras M.A., Perez L. Moses DT & H. Baldassarre fluorescent staining as a methods of assessing membrane damage and post-thawing survival of ram spermatozoa Theriogenology 41:483-489, 1994
[2] 曾培坚等.应用 MOET 繁育中国美利奴优质细毛羊.中国畜牧杂志,1997,3
[3] 杨永林等.不同超排方法对中国美利奴羊超排效果试验.中国养羊,1997,4
[4] 杨永林等.利用同期发情技术提高中国美利奴羊双胎率的试验.黑龙江动物繁殖,1998,1
[5] 倪健宏等.不同药物在绵羊同期发情中的应用试验.黑龙江动物繁殖,1997,2
[6] 杨永林等.中国美利奴羊非繁殖季节同期发情、冻胚移植试验.中国草食动物,1996,2
[7] 陈学进.最有前途的家畜育种方法.草食家畜 1996
[8] 刘志芬等.浅谈山羊冷配受胎率.黑龙江动物繁殖,2003,1
[9] 刘玉堂.精液冷冻保存的影响因素分析.黑龙江动物繁殖,2003,2
[10] 张国昌.羊人工授精站操作程序.黑龙江动物繁殖,2003,3
[11] 幸奠权.提高母羊繁殖力综合技术.黑龙江动物繁殖,2003,4
[12] 刘俊方 艾群等.羊的工人授精.黑龙江动物繁殖,2004,1
[13] 鞠连民.提高冷冻精液配种效果的体会.黑龙江动物繁殖,2004,4
[14] 刘孝德,姜勋平.山羊精液常温稀释液和最佳稀释液倍数研究.中国草食动物,2003,5
[15] 罗海玲,贾志海.维生素 E 对绵羊鲜精及冻精精液品质的影响.中国草食动物,2004,5
[16] 吴世林.用维生素 B 注射液作解冻液能提高情期受胎率.畜牧兽医杂志,1989 年第 1
[17] 吴永孝,郝正里.中式解冻液解冻牛精液的受胎试验.黄牛杂志,1995,20-21
[18].吴永孝等.牛冻精中药解冻液首次开发成功.中国农业科学,1992, 25(2)
[19] 新疆农垦绵羊冻精科研协作组.提高绵羊冷冻精液受胎率的研究.绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[20] 新疆自治区绵羊冻精技术研究协作组.绵羊冷冻精液技术试验研究报告(1975-1980).绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[21] 张春兰等.小尾寒羊精子内磷脂在冷冻一解冻前后含量的变化.黑龙江畜牧科技,1999,3
[22] 张坚中,陈元明等.全国绵羊冷冻精液技术中间试验北京片预备试验报告.绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[23] 张连峰等.冷冻前后牛羊精子表面三种凝集素受体的定量研究.中国农业科学,1994,27(1)
[24] 张文举,杨军祥等.谷肤甘肤提高牛冻精活力及受胎率的研究.甘肃畜牧兽医,1997 年 2
[25] 张扬.用 VB 作颗粒冻精解冻液提高母牛冻配受胎率的研究.草食家畜,1997,2
[26] 赵宗胜,代江生等.维生素 B 作鲜精稀释液提高绵羊产羔性能试验.草与畜杂志,1998,2
[27] 朱裕鼎,张坚中等.提高绵羊冷冻精液质量和受胎率的研究.绵羊精液冷冻资料汇编(全国绵羊精液冷冻技术科研协作组),1983
[28] 朱云彬.热滞蛋白在低温冷冻保存卵母细胞和胚胎中的作用.国外畜牧科技第 22 卷,2
[29] 安民家.畜精液的液态保存.家畜繁殖技术进展,1979,39
[30] Berger T,ally M.In vitro pertility evaluation ofcryoperserved ram semen and its corr Probnis E.Z., et al. Evaluation of relation fertility of cryopreserved goat sperm fertility of cryopreserved goat sperm. Theriogenology 41:711-717,1994
[31] Choudhry T. M.,Berger T.& D.elation with relative in vivo fertility.Theriogenology 44:1195-1200 1995
[32] Codde J,M&Berger T..In vivo fertilip, of rants in relation to sperm-zonapellucida binding and sperm-zona pellucida penetration of ovine oodles,Theriogenology 44:901-906, 1995
[33] Correa J.R&Zavos P.M.The h)poosrnotic swelling test: its employment as anassay to evaluate the functional integrity of the.f-ozen-thecnsed bovine spermmembrane Theriogenology 42:351-360,1994
[34] Correa J.R.&Zzvos P.M., Frozen-thawed bovine spermatozoa diluted,slowor rapid dilution method. measurements on occurrence of osmotic shock andsperm viability, Theriogenology 44:963-971, 1995
[35] Cross N.L..&Watson S.K., assessing acrosomal status of bovine sperm usingfluoresceinated lectins, Theriogenolo_y 42:89-98, 1994
[36] Deleeuw F.E. et al. Efects of various cryoprotective agents and membranestabilizing compounds on bull sperm membrane integrity after cooling andfreezing. Cryobiology, 1993, 30: 32-44
[37] Lenz R.W.. Ball G.D. Et Al. Chodroitin sulfate facilitates an acrosome reactionin bovine spermatozoa as evidenced by light microscopyelectron microscopy and vitro fertilization, Biol. Reprod. 1983; 28:683-690
[38] Mahaclevan M. et al. Efect of protective media and dilution methods。。preservation ofhuman spermatozoa. Andrologia, 1983, 15: 355-368
[39] Maxwell W.M.C., Landers A.J.&Evans G., Survival and fertility of ramspermatozoa frozen in pellets, straws and minitubes
[40] Molinia F. C., Evans G.&Maxwell, W.M.C., Incorporation of penetrating cryoprotectants in diluents for spermatozoa Theriogenology42:849-858,1994
[41] Maxwell W.M.C., Szell A., Hunton Jr,. Artificial breeding: embryotransfer and cloning. In: Oldham c.nt, martin gb, purvis iw (eds) reproduction ofmerino sheep-concepts and consequences. Ch. 17. Perth, School Of Agriculture(Animal Science), The University Of Western Australia, 1990; 217-237
[42] Nagy Sz,Hazas G.. Bali Papp A. et al. Evaluation of sperm tail menbraneintegrity 妙 the microscope Theriogenology 52:1153-1159,1999Polge C et al. Revival of spermatozoa after vitrification and dehydration at low temperature. Nature, 1949, 164:666
[43] Polge C et al. Revival of spermatozoa after vitrification and dehydration at low temperature. Nature, 1949, 164:666
[44] Sinha M.P.. Sinha A.K.&Singh B.K.. Effect of met 勿 Ixanthines of motility and fertility o f /cozen-thawed goat semen. Theriogenology 44:907-914. 1995
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