貉精液采集技术的研究
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摘要
哺乳动物精液的人工采集有重要的科学和应用价值，历来受到科学界的广泛关注。貉的生殖生理特性与其它家畜有较大的差异，这给貉人工采精技术的研究带来了很大的困难，目前在貉精液采集方面只有少量电刺激采精法成功报道。本研究以乌苏里貉为实验材料，采取电刺激法、假阴道法、按摩法三种方法对貉精液采集技术进行研究，以最终筛选出一种适合貉人工授精的最佳采精方法，主要内容包括：
(1)本实验采用电刺激法对13只貉进行了42次采精实验，成功采得38次，成功率为90.5％。貉每次射精量1.52±1.23ml，PH值6.56±0.37，密度5.43±1.08×10~8个／ml，活力0.9±0.2级，畸形率11.23±0.37％。
(2)本实验在分析貉的生理解剖和交配行为的基础上，设计了貉的假阴道，并在采精中进行了2次改进，初步建立了貉的假阴道采精器，在设计中改进了控制假阴道温度和压力的装置，温度用体温和水温共同来控制，压力用链球调节，这样既保证了假阴道温度长期的恒定，又实现了压力的自由调控；在外壳上安置了一个“锁结球”成功模拟了貉自然交配时的“锁结”行为。本实验虽未成功采得一次精液，但在貉采精方面首次尝试了假阴道采精法，为貉假阴道采精法的进一步研究提供了基本的技术参数。
(3)本实验用按摩法对貉精液采集进行了成功实验。在2006～2007年问共对57只貉进行了93次实验，成功采得26次，成功率为27.9％，并首次建立了貉按摩法的采精技术和精液品质参数。貉每次射精量0.29±0.18ml，PH值6.64±0.19，密度0.99±0.83×10~8个／ml，活力0.81±0.1，畸形率16.41±2.20％，活精子率85.11±8.3％。
在影响按摩法采精效果的诸多因素中，重点对刺激部位、训练、诱情、性腺的发育、选种五方面进行了实验分析。结果表明，貉射精的主要刺激部位在阴茎骨后缘两侧和龟头球下部锥性凹槽处；采精前对貉进行3～4天的训练，可消除貉对陌生环境的应激反应，增强貉在采精过程中和术者的配合程度；诱情尤其是放对诱情可明显增强貉的性兴奋，提高貉采精成功率；要提高貉采精成功率，从长远的角度考虑，通过配种记录和貉性腺的发育情况对貉进行选种是必要的。
结论：本实验研究认为，假阴道法采精目前是不成功的，假阴道的设计仍需改进；电刺激法虽成功率在90％以上，但由于操作复杂，对公貉伤害比较大，不适合用于貉人工授精；如果加强貉的采精训练和做好公貉的选种工作，按摩采精法可应用于貉人工授精。
Semen collection of mamal is extensive concerned in science, owning to importancevalue in science and the application.The raccoon dog is differrent with domestic animals inphysiology characteristics, This bring a very great difficulty in the research of semencollection of the raccoon dog.Electroejaculation have been reported a littile in semencollection of the raccoon dog. This research is taking three different ways, Artificial vagina,Electroejaculation, Massage method.Three kinds of methods collect a technique to carry onAI of the raccoon dog, the main contents include:
(1)The experiments were involved electroejaculation from 13 healthy raccoon dog whichwere 42 time to semen collection,During that time, the raccoon dog were ejaculated 38 times,which held 90.5 percent to the total number.The ejaculation volume of the raccoon dog was1.52±1.23ml; pH was 6.56±0.37, the sperm concentration was 5.43±1.08 million,Spermmotility was 91±0.2, The integrity rate of acrosome wasl 1.23±0.37%.
Electric voltage of ejaculation was 25～40V, Electric current frequency was 40～45Hz, the raccoon dog begin to ejaculate in 11～13min.The whole process is colected in 15～20min.
(2)This experiment have designed a Artificial vagina and improved 2 times in sememcollection, based on analysis of mating and the physiological dissection, one AV to collectsemen artificially was established initially.Some new data on the artificial vagina werepresented,one kind of new way to control the temperature has guaranted constantly vacationvagina temperature for long time, using the body temperature and water temperature tocontrol together.The pressure may adjustment freely through the chain ball;this experimenthave been imitated a mating behavior(lock knot) in nature, through having been placeda"lock knot ball" at outer shellof AV. Although this experiment didn't succeed to picked seminalfluid, it is the first research, on it and provide basic technical parameter for the furtherresearch in the AV of the raccoon dog.
(3)The experiments were involved Massage method from 57 healthy raccoon dog whichwere 96 time to semen Collection from 2006 to 2007. During that time, the raccoon dog wereejaculated 26 times, which held 27.9 percent to the total number. The ejaculation volumeof the raccoon dog was 0.29±0.18 ml. pH was 6.6±0.19, the sperm concentration was0.99±0.83 billion, Spermmotility was 0.9±0.2, The integrity rate of acrosome was16.41±2.20%,Rate of living was 85.11±8.3%.
The experiment analyzed mainly to five aspects in many factors, which effect results ofsemen collection,which includes the stimulated spot, the training, the incentive, the germ cells growth,and the animal selection.The result indicated, main stimulation spot to semination ofraccon, dog is in both sides behind penis bone,in channel place below Glans penisball.Restraining before semen collection from raccoon dog decrease stressTo new environmentand increases matching to holder for 3～4 day.The incentive stimuli are effective in increasingthe sexual response,and increase the number of sperm cells obtained.Considered from thelong-term angle, to enhance the fine success ratio of semen collection,collecting fine raccoondog is essential by sex glands and the breeding.record.
Conclusion: This study believed,artificial vagina is not successful in Semen collectionfrom the raccoon dog. The design to Artificial Vagina still had to be improved.Althoughelectroejaculation of the raccoon dog is above 90% in success ratio,it is not suitably in using tothe AI of the raccoon dog,because opratation is complexed,and it injury animalveryquitely.Massage Method is possible to apply in artificial insemination,if we selection fineseed and restraining before semen collection.
(1)本实验采用电刺激法对13只貉进行了42次采精实验，成功采得38次，成功率为90.5％。貉每次射精量1.52±1.23ml，PH值6.56±0.37，密度5.43±1.08×10~8个／ml，活力0.9±0.2级，畸形率11.23±0.37％。
(2)本实验在分析貉的生理解剖和交配行为的基础上，设计了貉的假阴道，并在采精中进行了2次改进，初步建立了貉的假阴道采精器，在设计中改进了控制假阴道温度和压力的装置，温度用体温和水温共同来控制，压力用链球调节，这样既保证了假阴道温度长期的恒定，又实现了压力的自由调控；在外壳上安置了一个“锁结球”成功模拟了貉自然交配时的“锁结”行为。本实验虽未成功采得一次精液，但在貉采精方面首次尝试了假阴道采精法，为貉假阴道采精法的进一步研究提供了基本的技术参数。
(3)本实验用按摩法对貉精液采集进行了成功实验。在2006～2007年问共对57只貉进行了93次实验，成功采得26次，成功率为27.9％，并首次建立了貉按摩法的采精技术和精液品质参数。貉每次射精量0.29±0.18ml，PH值6.64±0.19，密度0.99±0.83×10~8个／ml，活力0.81±0.1，畸形率16.41±2.20％，活精子率85.11±8.3％。
在影响按摩法采精效果的诸多因素中，重点对刺激部位、训练、诱情、性腺的发育、选种五方面进行了实验分析。结果表明，貉射精的主要刺激部位在阴茎骨后缘两侧和龟头球下部锥性凹槽处；采精前对貉进行3～4天的训练，可消除貉对陌生环境的应激反应，增强貉在采精过程中和术者的配合程度；诱情尤其是放对诱情可明显增强貉的性兴奋，提高貉采精成功率；要提高貉采精成功率，从长远的角度考虑，通过配种记录和貉性腺的发育情况对貉进行选种是必要的。
结论：本实验研究认为，假阴道法采精目前是不成功的，假阴道的设计仍需改进；电刺激法虽成功率在90％以上，但由于操作复杂，对公貉伤害比较大，不适合用于貉人工授精；如果加强貉的采精训练和做好公貉的选种工作，按摩采精法可应用于貉人工授精。
Semen collection of mamal is extensive concerned in science, owning to importancevalue in science and the application.The raccoon dog is differrent with domestic animals inphysiology characteristics, This bring a very great difficulty in the research of semencollection of the raccoon dog.Electroejaculation have been reported a littile in semencollection of the raccoon dog. This research is taking three different ways, Artificial vagina,Electroejaculation, Massage method.Three kinds of methods collect a technique to carry onAI of the raccoon dog, the main contents include:
(1)The experiments were involved electroejaculation from 13 healthy raccoon dog whichwere 42 time to semen collection,During that time, the raccoon dog were ejaculated 38 times,which held 90.5 percent to the total number.The ejaculation volume of the raccoon dog was1.52±1.23ml; pH was 6.56±0.37, the sperm concentration was 5.43±1.08 million,Spermmotility was 91±0.2, The integrity rate of acrosome wasl 1.23±0.37%.
Electric voltage of ejaculation was 25～40V, Electric current frequency was 40～45Hz, the raccoon dog begin to ejaculate in 11～13min.The whole process is colected in 15～20min.
(2)This experiment have designed a Artificial vagina and improved 2 times in sememcollection, based on analysis of mating and the physiological dissection, one AV to collectsemen artificially was established initially.Some new data on the artificial vagina werepresented,one kind of new way to control the temperature has guaranted constantly vacationvagina temperature for long time, using the body temperature and water temperature tocontrol together.The pressure may adjustment freely through the chain ball;this experimenthave been imitated a mating behavior(lock knot) in nature, through having been placeda"lock knot ball" at outer shellof AV. Although this experiment didn't succeed to picked seminalfluid, it is the first research, on it and provide basic technical parameter for the furtherresearch in the AV of the raccoon dog.
(3)The experiments were involved Massage method from 57 healthy raccoon dog whichwere 96 time to semen Collection from 2006 to 2007. During that time, the raccoon dog wereejaculated 26 times, which held 27.9 percent to the total number. The ejaculation volumeof the raccoon dog was 0.29±0.18 ml. pH was 6.6±0.19, the sperm concentration was0.99±0.83 billion, Spermmotility was 0.9±0.2, The integrity rate of acrosome was16.41±2.20%,Rate of living was 85.11±8.3%.
The experiment analyzed mainly to five aspects in many factors, which effect results ofsemen collection,which includes the stimulated spot, the training, the incentive, the germ cells growth,and the animal selection.The result indicated, main stimulation spot to semination ofraccon, dog is in both sides behind penis bone,in channel place below Glans penisball.Restraining before semen collection from raccoon dog decrease stressTo new environmentand increases matching to holder for 3～4 day.The incentive stimuli are effective in increasingthe sexual response,and increase the number of sperm cells obtained.Considered from thelong-term angle, to enhance the fine success ratio of semen collection,collecting fine raccoondog is essential by sex glands and the breeding.record.
Conclusion: This study believed,artificial vagina is not successful in Semen collectionfrom the raccoon dog. The design to Artificial Vagina still had to be improved.Althoughelectroejaculation of the raccoon dog is above 90% in success ratio,it is not suitably in using tothe AI of the raccoon dog,because opratation is complexed,and it injury animalveryquitely.Massage Method is possible to apply in artificial insemination,if we selection fineseed and restraining before semen collection.
引文
[1] 张荣祖等．中国哺乳动物分布．中国林业出版社．1997：420
[2] 赵正阶．中国东北地区珍稀濒危动物志 1999，609-612
[3] A. M. Mustonen, P. Nieminen ,M, Puukka ,J. Asikainen, S. Saarela, S.-L. Karonen, J. V. K. Kukkonen H. Hyvarinen. Physiological adaptations of the raccoon dog (Nyctereutes procyonoides) to seasonal fastingfat and nitrogen metabolism and influence of continuous melatonin treatment. J Comp Physiol B,2004,174: 1-12
[4] 佟煜人，钱国成主编．中国毛皮兽饲养技术大全．中国农业科技出版社．1990，1：6-10，22，261-278
[5] 华树芳．养貉业面临的形势及对策．特种经济动植物一2004年第7期
[6] 王百合，李成禹，王杰等．貉人工授精的初步研究．毛皮动物饲养．1988，2：15-16
[7] 张旭秋，许加成，郭淑芹等．貉精液冷冻配种技术的研究与效果．特种养殖．1999，4：5
[8] 王建辰，章孝荣主编．动物生殖调控．安徽科学技术出版社．1998，1：197-231，416-418
[9] 岳文斌，杨国义，任有蛇等．动物繁殖新技术．中国农业出版社．2003，1：16-22
[10] 徐旺生．我国近代家畜人工授精技术的引进和发展．古今农业．1996，1：29-34
[11] 潘永杰，贾世斋。野生动物人工采精方法探讨．动物学杂志．1998，33(5)：55-58
[12] 李涤非，吴晓民，李宝山等．乌苏里貉电刺激采精技术．经济动物学报．2003，7(3)：7～8
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[2] 赵正阶．中国东北地区珍稀濒危动物志 1999，609-612
[3] A. M. Mustonen, P. Nieminen ,M, Puukka ,J. Asikainen, S. Saarela, S.-L. Karonen, J. V. K. Kukkonen H. Hyvarinen. Physiological adaptations of the raccoon dog (Nyctereutes procyonoides) to seasonal fastingfat and nitrogen metabolism and influence of continuous melatonin treatment. J Comp Physiol B,2004,174: 1-12
[4] 佟煜人，钱国成主编．中国毛皮兽饲养技术大全．中国农业科技出版社．1990，1：6-10，22，261-278
[5] 华树芳．养貉业面临的形势及对策．特种经济动植物一2004年第7期
[6] 王百合，李成禹，王杰等．貉人工授精的初步研究．毛皮动物饲养．1988，2：15-16
[7] 张旭秋，许加成，郭淑芹等．貉精液冷冻配种技术的研究与效果．特种养殖．1999，4：5
[8] 王建辰，章孝荣主编．动物生殖调控．安徽科学技术出版社．1998，1：197-231，416-418
[9] 岳文斌，杨国义，任有蛇等．动物繁殖新技术．中国农业出版社．2003，1：16-22
[10] 徐旺生．我国近代家畜人工授精技术的引进和发展．古今农业．1996，1：29-34
[11] 潘永杰，贾世斋。野生动物人工采精方法探讨．动物学杂志．1998，33(5)：55-58
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