狐狸精液品质的研究
摘要
银狐(Vulpes falva)和蓝狐(Alopex lagopus)是珍贵的毛皮兽，其皮张是比较流行的裘皮原料。但目前我国人工饲养的狐品种退化，为此我国96年以来进口芬兰大体形蓝狐5000多只，并利用鲜精人工授精技术改良国内自繁普通蓝狐。可是，人们对芬兰蓝狐和改良一代蓝狐的精液特性还不很清楚，有必要对芬兰狐、改良狐、普通蓝狐和银狐的精液品质进行深入、细致的研究，同时探索狐精液冷冻工艺，延长精液的存活时间，提高公狐的利用率。本实验利用常规的精液晶质检查方法对芬兰狐、改良狐、普通蓝狐和银狐的精液品质进行了研究；对精子体外常湿保存活率和低渗肿胀试验(HOS)应用于狐精子生物学进行了探索：研究结果如下：
(1)3月下旬～4月上旬对4只芬兰狐采精21份，平均射精量、精子密度分别为0.69±0.18ml，1412.05±1157.70×10~6个／ml；对7只改良一代蓝狐采精53份，平均射精量，精子密度分别为0.71+0.37ml，1042.73±849.2×10~6个／ml；对10只普通蓝狐采精38份，平均射精量、精子密度分别为0.51±0.35ml、537.11±349.20×10~6个／ml；对9只银狐采精32份，平均射精量、精子密度分别为0.48+0.24ml、492.344±421.12×10~6个／ml。结果表明，芬兰狐和改良狐平均射精量、精子密度，即每次射精平均总精子数比普通蓝狐明显高，而普通银狐和蓝狐则类似。
(2)通过光学显微镜，利用线性测微尺测量芬兰狐、改良狐、普通蓝狐和银狐的长度，精子全长分别为66.23±1.2μm，65.30±1.42μm，65.34±1.35μm，67.31±2.79μm，头部长分别为6.75±0.17μm，6.78±0.22μm，6.64±0.22μm，6.64±0.23μm，中段长分别为12.33±0.35μm，12.29±0.32μm，12.56±0.68μm，12.84±0.74μm，主段长分别为43.13±0.72μm，42.76±2.00μm，42.37±0.63μm，43.84±1.61μm。结果表明，芬兰蓝狐、改良蓝狐和普通蓝狐精子长度相似，银狐和蓝狐精子长度没有明显差异。
(3)对蓝狐和银狐精液进行了低渗肿胀试验。结果表明，银狐和蓝狐精子尾部在低渗液下有9种类型的肿胀，其中有两种类型为尾部完全肿胀。将5只蓝狐平均活力为0.88的23份精液在低渗液中孵育150min以上，结果发现，尾部肿胀率在30min达到90％左右，90min达到最高值；而完全肿胀率在45min达到最高值。并且，对11份银狐和11份蓝狐精液低渗肿胀试验分析表明，尾部肿胀率、尾部完全肿胀率分别与活力显著正相关。为进一步用HOS试验评价狐精子受精力提供了材料和依据。
(4)本试验对30只进口芬兰大体型种公狐的112份精液于2000和2001年度两个繁殖季节进行了精子存活率的检测。结果表明：进口的芬兰大体型公狐人工授精的可利用率为83.4％；精子的平均总存活时间分别为A组26.0h，B组21.7h，C组5h：精子的平均生存指数分别为A组16.6，B组10.8，C组0.8；同组公狐中精子的存活率存在个体差别：同一个体公狐精子的存活率两个繁殖季节相对稳定。
The study on sperm biology of Finnish giant blue fox, general bule fox and hybird Fl (Finnish giant blue fox X general blue fox ) was conducted during 1998-1999.The study results were as follows) :
1. Semen sample were collected by digital manupulation. The ejaculate volume and sperm density were 6.69 + 0.8ml and 1412.05 + 1157.7.2x 106/ml for Finnish giant blue fox respectively; 0.71 +0.37ml and 1042.73 +849.2 X 106ml for hybrid F 1 ; 0.51 +0.35ml and 537.11 +349.7X 106/ml for general blue fox; 0.48 + 0.24ml and 492.34+421.12X 106/ml for silver fox. The average ejaculate volume and sperm density of Finnish giant blue fox and hybrid F 1 was higher than general blue fox and silver fox.
2. The sperm length of Finnish giant blue fox, hybrid Fl, general blue fox and silver fox was 66.23+1.21 um, 65.30+1.42 u m, 65.34 + 1.35 u m and 67.31 +2.79 u m respectively. The head length of sperm was 6.75 + 0.17 u m for giant blue fox, 6.78 + 0.22 u m for hybrid Fl, 6.64 + 0.22 um for general blue fox and 6.24 + 0.23 um for silver fox. The midpiece length of sperm tail was 12.33 + 0.35 u m for Finnish giant blue fox, 12.29 + 0.3211 m for hybrid Fl, 12.56 + 0.68urn for general blue fox and 12.84 + 0.74 u m for silver fox. The principle piece length of sperm tail was 43.13 + 0.72 u m for Finnish blue fox, 42.76 + 2 u m for hybrid Fl, 42.37 + 0.63 u m for general blue fox and 43.84+1.61 u m for silver fox. There was no significant difference in sperm length between Finnish giant blue fox, hybrid Fl, general blue fox and silver fox.
3. Nine type of sperm tail swelling were obseved in hypoosmotic swelling test, among them two type belonged to whole tail swelling, The percentage of sperm tail swelling and whole tail swelling showed a significant positive correlation to sperm motility.
4. Results showed that utilization ration of artificial insemination of import fmland fox was 83.4%. Mean survival time of A, B, C groups were 20.6h, 21.7hand5h, respectively. Mean survival index of sperm in A, B, C groups were 16.6, 10.8, 0.8, respectively.
(1)3月下旬～4月上旬对4只芬兰狐采精21份，平均射精量、精子密度分别为0.69±0.18ml，1412.05±1157.70×10~6个／ml；对7只改良一代蓝狐采精53份，平均射精量，精子密度分别为0.71+0.37ml，1042.73±849.2×10~6个／ml；对10只普通蓝狐采精38份，平均射精量、精子密度分别为0.51±0.35ml、537.11±349.20×10~6个／ml；对9只银狐采精32份，平均射精量、精子密度分别为0.48+0.24ml、492.344±421.12×10~6个／ml。结果表明，芬兰狐和改良狐平均射精量、精子密度，即每次射精平均总精子数比普通蓝狐明显高，而普通银狐和蓝狐则类似。
(2)通过光学显微镜，利用线性测微尺测量芬兰狐、改良狐、普通蓝狐和银狐的长度，精子全长分别为66.23±1.2μm，65.30±1.42μm，65.34±1.35μm，67.31±2.79μm，头部长分别为6.75±0.17μm，6.78±0.22μm，6.64±0.22μm，6.64±0.23μm，中段长分别为12.33±0.35μm，12.29±0.32μm，12.56±0.68μm，12.84±0.74μm，主段长分别为43.13±0.72μm，42.76±2.00μm，42.37±0.63μm，43.84±1.61μm。结果表明，芬兰蓝狐、改良蓝狐和普通蓝狐精子长度相似，银狐和蓝狐精子长度没有明显差异。
(3)对蓝狐和银狐精液进行了低渗肿胀试验。结果表明，银狐和蓝狐精子尾部在低渗液下有9种类型的肿胀，其中有两种类型为尾部完全肿胀。将5只蓝狐平均活力为0.88的23份精液在低渗液中孵育150min以上，结果发现，尾部肿胀率在30min达到90％左右，90min达到最高值；而完全肿胀率在45min达到最高值。并且，对11份银狐和11份蓝狐精液低渗肿胀试验分析表明，尾部肿胀率、尾部完全肿胀率分别与活力显著正相关。为进一步用HOS试验评价狐精子受精力提供了材料和依据。
(4)本试验对30只进口芬兰大体型种公狐的112份精液于2000和2001年度两个繁殖季节进行了精子存活率的检测。结果表明：进口的芬兰大体型公狐人工授精的可利用率为83.4％；精子的平均总存活时间分别为A组26.0h，B组21.7h，C组5h：精子的平均生存指数分别为A组16.6，B组10.8，C组0.8；同组公狐中精子的存活率存在个体差别：同一个体公狐精子的存活率两个繁殖季节相对稳定。
The study on sperm biology of Finnish giant blue fox, general bule fox and hybird Fl (Finnish giant blue fox X general blue fox ) was conducted during 1998-1999.The study results were as follows) :
1. Semen sample were collected by digital manupulation. The ejaculate volume and sperm density were 6.69 + 0.8ml and 1412.05 + 1157.7.2x 106/ml for Finnish giant blue fox respectively; 0.71 +0.37ml and 1042.73 +849.2 X 106ml for hybrid F 1 ; 0.51 +0.35ml and 537.11 +349.7X 106/ml for general blue fox; 0.48 + 0.24ml and 492.34+421.12X 106/ml for silver fox. The average ejaculate volume and sperm density of Finnish giant blue fox and hybrid F 1 was higher than general blue fox and silver fox.
2. The sperm length of Finnish giant blue fox, hybrid Fl, general blue fox and silver fox was 66.23+1.21 um, 65.30+1.42 u m, 65.34 + 1.35 u m and 67.31 +2.79 u m respectively. The head length of sperm was 6.75 + 0.17 u m for giant blue fox, 6.78 + 0.22 u m for hybrid Fl, 6.64 + 0.22 um for general blue fox and 6.24 + 0.23 um for silver fox. The midpiece length of sperm tail was 12.33 + 0.35 u m for Finnish giant blue fox, 12.29 + 0.3211 m for hybrid Fl, 12.56 + 0.68urn for general blue fox and 12.84 + 0.74 u m for silver fox. The principle piece length of sperm tail was 43.13 + 0.72 u m for Finnish blue fox, 42.76 + 2 u m for hybrid Fl, 42.37 + 0.63 u m for general blue fox and 43.84+1.61 u m for silver fox. There was no significant difference in sperm length between Finnish giant blue fox, hybrid Fl, general blue fox and silver fox.
3. Nine type of sperm tail swelling were obseved in hypoosmotic swelling test, among them two type belonged to whole tail swelling, The percentage of sperm tail swelling and whole tail swelling showed a significant positive correlation to sperm motility.
4. Results showed that utilization ration of artificial insemination of import fmland fox was 83.4%. Mean survival time of A, B, C groups were 20.6h, 21.7hand5h, respectively. Mean survival index of sperm in A, B, C groups were 16.6, 10.8, 0.8, respectively.
引文
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71． Crozet, N. (1984) , Vltraqtructural aspects of in vizo fevtilization in the cow. Gamete Res. 10:241-251．
72． Cummins, J.M. and woodall. P.P. (1985) On mammalian sperm dimensions. J. Reprod. Pert. 75, 153-175．
73． Cummins, J.M. (1983) Sperm size, body mass and reproduction in mammals.In The Sperm Cell, PP.395-398． Ed.J.Andre Martimus Nijhoff, The Hague.
74． Dandekar P, Talbot P. (1992) Perivitelline space of mammalian oocytes: Extracellular matrix of unfertilized oocytes and formation of a cortical granule envelope following fertilization. Mol Reprod Dev. 31 (2) :135．
75． Davis, M.L. (1982) The spermatozoon of the common Marmoset, Callithrix jacchus (Primates, Callithricidae ) :an ultrastructural investigation. Ph.D. thesis , Texas A & M University. (Diss.Abstr.Int.43B 1696-1697)
76． De Loos F, Van Maurik P, Van Beneden T, et al (1992) , Structural aspects of bovine oocyte maturation in vitro. Mol Reprod Dev. 31 (3) :208．
77． Ding JC, Clarke N, Nagai T, Moor RM. (1992) , Protein and nuclear changes in pig eggs at fertilization. Mol Reprod Dev. 31 (4) :287．
78． Dobrinski, I., Lulai, C., Barm, A.D.andPost, K. (1993) Effects of four different extenders and three different freezing rates on post thaw viability of dog semen. J. Reprod. Fertil. Suppl., 47:291-296．
79． Dott, H.M.&Utsi, M.N.P. (1971) The collection and examination of semen of the Reindeer (Rangifer tarandus) J.Zool., Lond.164, 419-424．
80． EI-Gaafary．M．N．等著，单志清摘择(1991) ．国外畜牧科技．(2) ：21-22．
81． Elder, F.F.B.Hsu.T.C. (1981) Silver staining patterns of mammalian epididymal spermatozoa. Cytogenet. cell Genet. 30, P157-167
82． England, G.C.W. (1992) The cryopreservation of dog semen. Thesis, Royal Veterinary College, University of London, UK, 152pp.
83． Farstad, w.K.., Fougner, J. A. and Andersen Berg, K. (1992c) Species differences in fertility after artificial insemination with frozen semen in fox pure breeding. Norw. J. Agric. Sci. , Suppl. , 9:115-121．
84． Farstad, W (1996) seman cryopreservation in dogs and foxes. Auim. Reprod. Sci. 42, 251-260．
85． Farstad, W. ( 1984) Bitch fertility after natural mating and after artificial insemination with fresh or frozen semen. J. Small Anim. Pract., 25:561-565．
86． Farstad, W., Fougner, J.A. and Toues, C.G. (1992b) The optimum time for artificial insemination of blue fox vixens (Alopex lagopus) with frozen-thawed semen from silver foxes ( Vulpes vulpes ) semen. Theriogenology, 38:853-856．
87． Farstad, W., Fougner. J.a. and Torres. C.G (1992a) The effect of sperm number on fertility in blue fox vixens (Alopex lagopus) artificially inseminated with frozen silver fox (Vulpes vulpes) semen. Theriogenology, 36:699-711．
88． Farstad, W., Krogen s, A., Nagyova, E., Hafne, A.L. and Hyttel, P., (1993) In vitro techniques in fox reproduction .Livest. Prod.Sci., 36:23-27．
89． Fawcett, D.W. (1958) The structure of the mammalian spermatozoon, int. Rev. Cytol. 7, 195-235
90． Fawcett, D.W. (1970) A comparative view of spermatozoa ultrastructure.Biol. Reprod (Suppl.) 2:90-127．
91． Ogawa, k, etal (1977) Identification of dynein as the outer arms of se urchin sperm axonemes. Matl. Acad. Sci. USA. 74:5006-5010．
92． Fawcett, D.W. (1975) The mammalian spermatozoon. Deveiop.Biol. 44:394-436．
93． Fawcett, D.W. (1976) Unsolved problems in morphogenesis of the mammalian spermatozoon. In International Cell Biology 1976-1977, PP.288-601． EdsB. R. Brinkley & K.R.Porter.Rockefeller U.Press.Nwe York.
94． Fawcett. D.W. (1975) The mammalian spermatozoon. Dev. Biol.44:394．
95． Fawcett.D.W (1975) The mammalinan spermatozoan Dev. Biol..44:394-436．
96． Fawcett.D.W. Anderson, W.A.& Phillips, D.M. (1971) Morphogenetic factors influencing the shape of the sperm head. Devi Biol .26 , 220-251
97． Feito, R.&Gallardo, M. (1976) Notes on the sperm morphology of Ctenomys maulinus (Rodentia, Octodontidae) Experientia 32, 734-735．
98． Feito, R.& Gallardo , M. (1982) Sperm morphology of the Chilean species of Ctenomys (Octodontidae) J.Mammal.63 , 658-661．
99． Flechon, J.E.& Hafez, E.S.E. (1976) Scanning electron microscopy of human spermatozoa. In Human Semen and Fertility Regulation in Man, pp.76-82． Ed.E.S.E.Hafez.C.V.Mosby Co, St Louis.
100． Fleming, A.D., Yanagimachi, R.& Yanagimachi, H. (1981) Spermatozoa of the Atlantic bottlenosed dolphin, Tursiops truncatus. J. Reprod. Pert. 63, 509-514．
101． Fontbonne, A. and Badinand, F. (1993) Canine artificial insemination with frozen semen:comparison of intravaginal and intrauterine deposition of seme. J. Reprod. Fertil. Suppl., 47:325-327．
102． Foote, R.H., 1964． Extenders for freezing dog semen.Am.J.Vet.Res., 25:37-40．
103． Forman, G.L. (1968) Comparative gorss morphology of spermatozoa of two families of North American bats. Univ.Kansas Sci.Bull. 47 (No.16) , 901-908
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60． Bwanga, C.O. (1991) Cryopreservation of boar semen.In:A literature review. Acta Vet.Scand., 32:431-453．
61． Calow, P. (1979) The cost of reproduction-a physiological aproach.Biol.Rev.54, 23-40．
62． Chandley, A.C., Jones, R.C.Dott, H.M., Allen, W.R.& Short, R.V. (1974) Meiosis in interspecific
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63． Charlesorth, B. (1978) The population genetics of anisogamy, J.theor, Biol.73, 347-357．
64． Chittleborough, R.G.( 1995) Aspects of reproduction in the male Humpback whale, Megaptera nodosa (Bonaterre) .Aust.J.mar.Freshwater Res.6, 1-29．
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66． Could, K.G., Martin, D.E.& Hafez.E.S.E. (1975) Mammalian spermatozoa. In SEM atlas of Mammalian Reproduction, PP.42-57． Ed.E.S.E. Hafez. Springer-Verlag, New York.
67． Courteas, J.L., and paquighon, M. (1985) Ultrastrcuture of fresh, and frozen, and frozen-thawed spermatozoa of the boar. In "Deep Freezing of BOAR Semen", PP. 61-99． Swedish university on Agacutural Sciences. Uppsala.
68． Couturier, M.A.J. (1956) La spermatozoide du Buoquetin des Alpes (Capra ibex ibex L. 1758) , Mammalia 20, 124-127．
69． Cran DG, Cheng WTK. (1985) , Changes in cortical granules during prcine oocyte maturation. Gamete Res.11 (3) :311．
70． Cran DG, Cheng WTK. (1986) , The cortical reaction in pig oocytes during in vivo and in vitro fertilization. Gamete Res. 13 (3) :241
71． Crozet, N. (1984) , Vltraqtructural aspects of in vizo fevtilization in the cow. Gamete Res. 10:241-251．
72． Cummins, J.M. and woodall. P.P. (1985) On mammalian sperm dimensions. J. Reprod. Pert. 75, 153-175．
73． Cummins, J.M. (1983) Sperm size, body mass and reproduction in mammals.In The Sperm Cell, PP.395-398． Ed.J.Andre Martimus Nijhoff, The Hague.
74． Dandekar P, Talbot P. (1992) Perivitelline space of mammalian oocytes: Extracellular matrix of unfertilized oocytes and formation of a cortical granule envelope following fertilization. Mol Reprod Dev. 31 (2) :135．
75． Davis, M.L. (1982) The spermatozoon of the common Marmoset, Callithrix jacchus (Primates, Callithricidae ) :an ultrastructural investigation. Ph.D. thesis , Texas A & M University. (Diss.Abstr.Int.43B 1696-1697)
76． De Loos F, Van Maurik P, Van Beneden T, et al (1992) , Structural aspects of bovine oocyte maturation in vitro. Mol Reprod Dev. 31 (3) :208．
77． Ding JC, Clarke N, Nagai T, Moor RM. (1992) , Protein and nuclear changes in pig eggs at fertilization. Mol Reprod Dev. 31 (4) :287．
78． Dobrinski, I., Lulai, C., Barm, A.D.andPost, K. (1993) Effects of four different extenders and three different freezing rates on post thaw viability of dog semen. J. Reprod. Fertil. Suppl., 47:291-296．
79． Dott, H.M.&Utsi, M.N.P. (1971) The collection and examination of semen of the Reindeer (Rangifer tarandus) J.Zool., Lond.164, 419-424．
80． EI-Gaafary．M．N．等著，单志清摘择(1991) ．国外畜牧科技．(2) ：21-22．
81． Elder, F.F.B.Hsu.T.C. (1981) Silver staining patterns of mammalian epididymal spermatozoa. Cytogenet. cell Genet. 30, P157-167
82． England, G.C.W. (1992) The cryopreservation of dog semen. Thesis, Royal Veterinary College, University of London, UK, 152pp.
83． Farstad, w.K.., Fougner, J. A. and Andersen Berg, K. (1992c) Species differences in fertility after artificial insemination with frozen semen in fox pure breeding. Norw. J. Agric. Sci. , Suppl. , 9:115-121．
84． Farstad, W (1996) seman cryopreservation in dogs and foxes. Auim. Reprod. Sci. 42, 251-260．
85． Farstad, W. ( 1984) Bitch fertility after natural mating and after artificial insemination with fresh or frozen semen. J. Small Anim. Pract., 25:561-565．
86． Farstad, W., Fougner, J.A. and Toues, C.G. (1992b) The optimum time for artificial insemination of blue fox vixens (Alopex lagopus) with frozen-thawed semen from silver foxes ( Vulpes vulpes ) semen. Theriogenology, 38:853-856．
87． Farstad, W., Fougner. J.a. and Torres. C.G (1992a) The effect of sperm number on fertility in blue fox vixens (Alopex lagopus) artificially inseminated with frozen silver fox (Vulpes vulpes) semen. Theriogenology, 36:699-711．
88． Farstad, W., Krogen s, A., Nagyova, E., Hafne, A.L. and Hyttel, P., (1993) In vitro techniques in fox reproduction .Livest. Prod.Sci., 36:23-27．
89． Fawcett, D.W. (1958) The structure of the mammalian spermatozoon, int. Rev. Cytol. 7, 195-235
90． Fawcett, D.W. (1970) A comparative view of spermatozoa ultrastructure.Biol. Reprod (Suppl.) 2:90-127．
91． Ogawa, k, etal (1977) Identification of dynein as the outer arms of se urchin sperm axonemes. Matl. Acad. Sci. USA. 74:5006-5010．
92． Fawcett, D.W. (1975) The mammalian spermatozoon. Deveiop.Biol. 44:394-436．
93． Fawcett, D.W. (1976) Unsolved problems in morphogenesis of the mammalian spermatozoon. In International Cell Biology 1976-1977, PP.288-601． EdsB. R. Brinkley & K.R.Porter.Rockefeller U.Press.Nwe York.
94． Fawcett. D.W. (1975) The mammalian spermatozoon. Dev. Biol.44:394．
95． Fawcett.D.W (1975) The mammalinan spermatozoan Dev. Biol..44:394-436．
96． Fawcett.D.W. Anderson, W.A.& Phillips, D.M. (1971) Morphogenetic factors influencing the shape of the sperm head. Devi Biol .26 , 220-251
97． Feito, R.&Gallardo, M. (1976) Notes on the sperm morphology of Ctenomys maulinus (Rodentia, Octodontidae) Experientia 32, 734-735．
98． Feito, R.& Gallardo , M. (1982) Sperm morphology of the Chilean species of Ctenomys (Octodontidae) J.Mammal.63 , 658-661．
99． Flechon, J.E.& Hafez, E.S.E. (1976) Scanning electron microscopy of human spermatozoa. In Human Semen and Fertility Regulation in Man, pp.76-82． Ed.E.S.E.Hafez.C.V.Mosby Co, St Louis.
100． Fleming, A.D., Yanagimachi, R.& Yanagimachi, H. (1981) Spermatozoa of the Atlantic bottlenosed dolphin, Tursiops truncatus. J. Reprod. Pert. 63, 509-514．
101． Fontbonne, A. and Badinand, F. (1993) Canine artificial insemination with frozen semen:comparison of intravaginal and intrauterine deposition of seme. J. Reprod. Fertil. Suppl., 47:325-327．
102． Foote, R.H., 1964． Extenders for freezing dog semen.Am.J.Vet.Res., 25:37-40．
103． Forman, G.L. (1968) Comparative gorss morphology of spermatozoa of two families of North American bats. Univ.Kansas Sci.Bull. 47 (No.16) , 901-908