Return to search

Efici?ncia da congela??o automatizada na viabilidade de s?men bovino. / Efficiency of automated freezing on the viability of bovine semen.

Submitted by Sandra Pereira (srpereira@ufrrj.br) on 2017-07-24T12:38:24Z
No. of bitstreams: 1
2010 - Wilson Franklim de Vasconcelos Filho.pdf: 785209 bytes, checksum: a1bf8c5a2306da3cdf30db3da6cf4f0d (MD5) / Made available in DSpace on 2017-07-24T12:38:24Z (GMT). No. of bitstreams: 1
2010 - Wilson Franklim de Vasconcelos Filho.pdf: 785209 bytes, checksum: a1bf8c5a2306da3cdf30db3da6cf4f0d (MD5)
Previous issue date: 2010-03-05 / Planos de Reestrutura??o e Expans?o das Universidades Federais (Reuni) / Even with technological advances in the field of artificial insemination, there is a huge
number of people freezing semen in a manual way. This technique has proven to be viable,
however, it is difficult to standardize the cooling and freezing curves, since they depend on
the quality of the material used (refrigerator sealing, type of cooler, liquid nitrogen level and
other). Therefore, the objective of this study was to evaluate the effect of the cryopreservation
of bovine semen using two freezing techniques (conventional and automated) on motility,
vigor and heat resistance of the sperm after thawing. The experiment was developed over a
period of 12 weeks, divided into two parts, an initial six weeks part where Tris-yolk extender
was used and a second part in which the extender used was Citrate-yolk. Three Holstein bulls
were used for weekly semen collection by artificial vagina method. After collection, were
evaluated: volume, motility, mass motility and sperm concentration to find out the number of
doses. After dilution, the semen was stored in straws of 0.5 ml. Then the straws were
subjected to conventional freezing technique (use of refrigerator, nitrogen vapor and
immersion in liquid nitrogen) and automated technique (machine from Cryogen ?). After 24
hours the samples were thawed, re-tested and underwent the quick thermoresistance test. The
results were analyzed by Kruskal-Wallis analysis of variance (nonparametric) with a
significance level of 5% with the aid of BioEstat 4 and Systat 11 PC-programs. There was
significant difference (p <0.05) between automated and manual techniques for both extenders
used, and the automated technique has presented the best results for motility and vigor after
cryopreservation and thermoresistance test. Thus, we can conclude that the automated
technique to freezing bovine semen has shown to be advantageous and practical because as
well as providing better results, it standardizes cooling and freezing curves. / Mesmo com os avan?os tecnol?gicos no campo da insemina??o artificial, ainda ? grande o
n?mero de pessoas que realizam a congela??o de s?men de maneira manual. Esta t?cnica tem
se mostrado vi?vel, por?m, ? dif?cil padronizar as curvas de resfriamento e de congela??o,
uma vez que dependem da qualidade do material utilizado (veda??o da geladeira, tipo de
caixa de isopor, quantidade de gelo, n?vel de nitrog?nio l?quido e outros). Portanto, o objetivo
deste estudo foi avaliar o efeito da criopreserva??o de s?men bovino utilizando duas t?cnicas
de congela??o (convencional e automatizada) sobre a motilidade, vigor e resist?ncia t?rmica
dos espermatoz?ides ap?s a descongela??o. O experimento foi elaborado em um per?odo de
12 semanas, sendo dividido em duas partes, uma inicial de seis semanas onde foi utilizado o
diluidor Tris-gema e uma segunda parte na qual o diluidor utilizado foi Citrato-gema. Foram
utilizados tr?s touros holandeses para coleta de s?men semanal pelo m?todo da vagina
artificial. Ap?s a coleta, foram avaliados: volume, motilidade, turbilhonamento, vigor e
concentra??o esperm?tica para determina??o do n?mero de doses. Ap?s dilui??o, o s?men foi
envasado em palhetas de 0,5 ml. Em seguida, as palhetas foram submetidas ?s t?cnicas de
congela??o convencional (uso de geladeira, vapor de nitrog?nio e imers?o em nitrog?nio
l?quido) e automatizada (aparelho da Cryogen?). Ap?s 24 horas as amostras foram
descongeladas, reavaliadas e submetidas ao teste de termoresist?ncia r?pido (TTR). Os
resultados foram analisados pelo teste Kruskal-Wallis (an?lise de vari?ncia n?o param?trica)
com n?vel de signific?ncia de 5% com aux?lio dos programas BioEstat 4 e Systat 11. Houve
diferen?a significativa (p<0,05) entre as t?cnicas automatizada e manual para os dois
diluidores testados, sendo a t?cnica automatizada a que apresentou melhores resultados para
motilidade e vigor ap?s a criopreserva??o e teste de resist?ncia t?rmica. Desta forma, pode-se
concluir que a t?cnica automatizada para congela??o de s?men bovino apresenta-se vantajosa
e pr?tica pois al?m de proporcionar melhores resultados, ela padroniza as curvas de
refrigera??o e de congelamento.

Identiferoai:union.ndltd.org:IBICT/oai:localhost:jspui/1903
Date05 March 2010
CreatorsVasconcelos Filho, Wilson Franklim de
ContributorsMello, Marco Roberto Bourg de, Dias, Angelo Jos? Burla, Jesus, Vera L?cia Teixeira de
PublisherUniversidade Federal Rural do Rio de Janeiro, Programa de P?s-Gradua??o em Zootecnia, UFRRJ, Brasil, Instituto de Zootecnia
Source SetsIBICT Brazilian ETDs
LanguagePortuguese
Detected LanguageEnglish
Typeinfo:eu-repo/semantics/publishedVersion, info:eu-repo/semantics/masterThesis
Formatapplication/pdf
Sourcereponame:Biblioteca Digital de Teses e Dissertações da UFRRJ, instname:Universidade Federal Rural do Rio de Janeiro, instacron:UFRRJ
Rightsinfo:eu-repo/semantics/openAccess
RelationAGCA, Y.; CRITSER, J. K.; Cryopreservation of spermatozoa in assited reproduction. Seminars in Reproductive Medicine, v. 20, n. 1,p. 15-23, 2002. AGUIAR, P. R. L.; MORAES, J. B.; MALSCHITZKY, E.; SILVA, A. C. Criopreserva??o de s?men bovino utilizando diluente ? base de PBS com tr?s diferentes percentuais de gema de ovo. Veterin?ria em Foco, Canoas, v. 5, n. 1, p. 44-50, 2007. AIRES, V.A.; HINSCH, K.D.; SCHLOESSER, F.M. et al. In vitro and in vivo comparison of egg yolk-based and soybean lecithun-based extenders for cryopreservation of bovine semen. Theriogenology, v.60, p.269-279, 2003. ALMQUIST, J.O.; WIGGINS, H. B. Survival of bull spermatozoa frozen and thawed by different methods in plastic straws. A I Digest, v. 21, n. 7, p. 12, 1973. AMANN, R.P., PICKETT, B.W. Principals of cryopreservation and a review of cryopreservation of stallion spermatozoa, Journal of Equine Veterinary Science, v.7, p.145- 173, 1987. AMIRAT, L.; TAINTURIER, D.; JEANNEAU, L.; THORIN, C.; GERARD, O. COURTENS, L.J.; ANTON, M. Bull semen en vitro fertility after cryopreservation using egg yolk LDL: a comparison with Optidyl?, a commercial egg yolk extender. Theriogenology, v. 61, p.895-907, 2004. ARRUDA, R. P. Avalia??o dos efeitos de diluidores e crioprotetores para o espermatoz?ide eq?ino pelo uso de microscopia de epifluoresc?ncia, citometria de fluxo, an?lises computadorizadas da motilidade (CASA) e da morfometria (ASMA). 2000. 121f. Tese (Livre Doc?ncia) - Faculdade de Medicina Veterin?ria e Zootecnia, Universidade de S?o Paulo, S?o Paulo, 2000. ARRUDA, R. P.; ANDRADE, A. F. C.; PERES, K. R.; RAPHAEL, C. F.; NASCIMENTO, J.; CELEGHINI, E. C. C. Biot?cnicas aplicadas ? avalia??o do potencial de fertilidade do 46 s?men eq?ino. Revista Brasileira de Reprodu??o Animal, Belo Horizonte, v.31, n.1, p.8-16, jan./mar. 2007. ARRUDA, R. P.; BALL, B. A.; GRAVANCE, C. G.; LIU, I. K. M. Determina??o da integridade da membrana plasm?tica e acrossomo de espermatoz?ides de garanh?es pela t?cnica de citometria de fluxo. Acta Scientiae Veterinariae, v.31, p.226- 227, 2003. ARRUDA, R.P.; CELEGHINI, E.C.C. Valida??o de uma t?cnica para avalia??o simult?nea das membranas plasm?tica, acrossomal e mitocondrial de espermatoz?ides bovinos. Acta Scientiae Veterinariae, v.31, p.230-231, 2003. ASBIA (Associa??o Brasileira de Insemina??o Artificial). Dispon?vel em <HTTP://www.asbia.org.br/>. Acesso em 7 de Julho de 2009. ASHWOOD-SMITH, M.J. Mechanisms of cryoprotectant action. In: BOWLER, K., FULLER, B.J. Temperature and animal cels. Cambridge: The Co. of Biologists Ltd. P.395- 406, 1986. BALL, B. A. Na introduction to the use and implication of cryopreserved equine semen. In: EQUINE ASSISTED REPRODUCTIVE TECNOLOGY WORKSHOP, Davis, CA, 1998. Proceedings. Davis: School of Veterinary Medicine University of California, 1998. p. 25-43. BARROS, P. M. H. Estresse oxidativo e integridade do DNA em s?men resfriado de gato-domato- pequeno (Leopardus tigrinus, SCHREBER, 1775). Tese (doutorado) ? Universidade de S?o Paulo. Faculdade de Medicina Veterin?ria e Zootecnia. 120 f. 2007 BECKER, W.C., SENGER, P.L., AALSETH, E.P. Influence of glycerol levels, diluent and post-thaw temperature on motility and acrossomal maintenance in bovine semen frozen in plastic straws. Journal of Animal Science, v.44, p.1067-1071, 1977. BUCAK, M.N, ATESSAHIN, A., Y?CE, A. Effect of anti-oxidants and oxidative stress parameters on ram semen after the freeze-thawing process. Small Ruminant Research, v.75, p.128-134, 2008. 47 CASEY, P. J.; HILLMAN, R. B.; ROBERTSON, K. R.; YUDIN, A. I.; LIU, I. K. M.; DROBINS, E. Z. Validation of an acrosomal stain for equine sperm that differentiates between living and dead sperm. Journal of Andrology, v.14, p.289-297, 1993. CASTELO, T. S.; FROTA, T. R.; SILVA, A. R. Considera??es sobre a criopreserva??o do s?men de caprinos. Acta Veterinaria Brasilica, v.2, n.3, p.67-75, 2008. CELEGHINI, E. C. C. Efeitos da criopreserva??o do s?men bovino sobre as membranas plasm?tica, acrossomal e mitocondrial e estrutura da cromatina dos espermatoz?ides utilizando sondas fluorescentes. 2005. 186f. Tese (Doutorado) - Faculdade de Medicina Veterin?ria e Zootecnia, Universidade de S?o Paulo, S?o Paulo, 2005. CELEGHINI, E. C. C.; ARRUDA, R. P.; ANDRADE, A.F.C.; RAPHAEL, C. F.; NASCIMENTO, J. Simultaneous evaluation of the plasmatic, acrosomal and mitochondrial membranes in equine spermatozoa. In: International Congress of Animal Reproduction, 15, 2004, Porto Seguro. Abstract ... Belo Horizonte: CBRA, 2004. p.511. Resumo. CHERNOS, J.E.; MARTIN, R.H. A cytogenetic investigation of the effects of cryopreservation on human sperm. American Journal of Humam Genetics, v. 45, p. 766? 777, 1989. CONCANNON, P.W., BATTISTA, M. Canine semen freezing and artificial insemination. IN: KIRK. Current veterinary therapy ? Small animal practice, X. Philadelphia : Saundrs, 1989. p.l247-1289. CROSS, N. L.; MORALES, P.; OVERSTREET, J. W.; HANSON, F. W. Two simple methods for detecting acrosome ? reacted human sperm . Gamete Research, v.15, p.213-226, 1986. DALIMATA, A.M., GRAHAM, J.K. Criopreservation of rabbit spermatozoa using acetamide in combination with trehalose and methyl cellulose. Theriogenology, v.48, p.831-841, 1997. DARIN-BENNETT, A., WHITE, I.G. Influence of the cholesterol content of mammalian spermatozoa on susceptibility to cold-shock. Cryobiology, v.14, p.466?470, 1977. 48 De LEEW, F. E.; CHEN, H. C.; COLENBRANDER, B.; VERKLEIJ, A. J. Cold-induced ultrastructural changes in bull and boarsperm plasma membranes. Cryobiology, v. 27, p. 171- 183, 1990. De LEEW, F. E.; De LEEW, A. M.; DEN DAAS, J. H. G.; COLENBRANDER, B.; VERKLEIJ, A.J. Effects of various cryoprotective agents and membrane-stabilizing compounds on bull sperm membrane integrity after cooling and freezing. Cryobiology, v. 30, p. 32-44, 1993. DIMITROPOULOS R. La signification du test de la thermor?sistance dans l?ppreciation de la valeur f?condant du sperma congel?. Animal Medicin Veterinary, v.4, p.215-24, 1967. ENGLAND, J.; SCARLETT, J.; MEYERS-VALEN et al. Computer-assisted sperm analysis of canine spermatozoa motility mesasurements. Theriogenology, v. 40, p.725-733, 1993. EVANS, G.; MAXWELL, W.M.C. Salamon?s. Artificial insemination of sheep and goats. Butterworth, Sydney 1987. EVENSON, D.P.; DARZYNKIEWICZ, Z.; MELAMED, M.R. Relation of mammalian sperm chromatin heterogeneoty to fertility. Science, v. 210, p.1131?1133, 1980. EVENSON, D.P.; JOST, L. Sperm chromatin structure assay: DNA denaturability. Methods Cell Biol, v. 42, p. 159?175, 1994. FAHY, G.M. The relevance of cryoprotectant ?toxicity? to cryobiology. Cryobiology, v.23, p.1-13, 1986. FARLIN, M. E.; JASKO, D. J.; GRAHAM, J. K.; SQUIRES, E. L. Assessment of Pisum sativum agglutinin in identifying acrosomal damage in stallion spermatozoa. Molecular Reproduction Development, v.32, p.23-27, 1992. FISER, P. S.; FAIRFULL, R. W. The effect of glycerol concentration and cooling velocity on cryosurvival of ram spermatozoa frozen in straws. Cryobiology, v. 21, p. 542-551, 1984. 49 FOOTE, R. H.; L. C. GRAY; YOUNG, D. C.; DUNN, H. O. Fertility of bull semen stored up to four days at 5?C in 20% egg yolk extenders. Journal of Dairy Science, v. 43, n. 9, p. 1330- 1334, 1960. FOOTE, R. H. Influence of extender, extension rate, and glycerolating tecnique on fertility of frozen bull semen. Journal Dairy Science, v. 53, p. 1478-1482, 1970. FOOTE, R.H.; BROCKETT, C.C.; KAPROTH, MT. Motility and fertility of bull sperm in whole milk extender containing antioxidants. Animal Reproduction Science., v. 71, p.13-23, 2002. GARNER D. L.; JOHNSON L. A. Viability assessment of mammalian sperm using SYBR-14 and propidium iodide. Biology of Reproduction, v.53, p.276-284, 1995. GARNER, D. L.; THOMAS, C. A.; GRAVANCE, C. G. Seminal plasma addition attenuates the dilution effect in bovine sperm. Theriogenology, v. 56, p.31-40, 2001. GARNER, D. L.; THOMAS, A. C.; GRAVANCE, C. G. The effect of glycerol on the viability, mitochondrial function and acrosomal integrity of bovine spermatozoa. Reproduction of Domestic Animals, v.34, p.399-404, 1999. GONZALEZ, R. A. F. Efeito da criopreserva??o usando diferentes t?cnicas de congela??o e crioprotetores sobre par?metros esperm?ticos e a integridade de membranas do espermatoz?ide bovino. 2004. 92f. Tese (doutorado). Faculdade de Medicina Veterin?ria e Zootecnia, Universidade de S?o Paulo, Pirassununga, 2004. GRAHAM J.K.; KUNZE E.; HAMMERSTEDT R. H. Analysis of sperm cell viability, acrosomal integrity, and mitochondrial funtion using flow citometry. Biology of Reproduction, v.43, p.55-64, 1990. GRAVES, W. M. Improving Artificial Insemination Techniques. Bulletin 1325, University of Georgia, Cooperative Extension. p.1-6, 2007. 50 GUTHRIE, H.D.; LIU, J.; CRITSER, J. K. Osmotic tolerance limits and effects of cryoprotectants on motility of bovine spermatozoa. Biology of Reproduction, v. 67, p. 1811-1816, 2002. HAFEZ, E. S. E.; HAFEZ, B. Reprodu??o Animal. Editora Manole. 7?ed. p. 381-470. 2004. HALANGK, W.; FRANK, K., BOHNENSACK, R. Zur bestimmung der menge intakter spermien in bullenejakulaten. Arch Exp Vet Med, v.38, p.105-114, 1984. HAMMERSTEDT, R.H.; GRAHAM, J.K. Cryopreservation of poutry sperm: The enigma of glycerol. Cryobiology, v.29, p. 26-38, 1992. HARRISON, R. A. P.; VICKERS, S. E. Use of fluorescent probes to assess membrane integrity in mammalian spermatozoa. Journal of Reproduction and Fertility, v.88, p.343- 352, 1990. HOLT, W. V. Fundamental aspects of sperm cryobiology: the importance of species and individual differences. Theriogenology, Montgomery, v. 53, p. 47-58, 2000. JEYENDRAN, R.S., GRAHAM, E.F. An evaluation of cryoprotective compounds on bovine spermatozoa. Cryobiology, v.17, p.458-464, 1980. KARABINUS, D.S., EVENSON, D.P., KAPROTH, M.T. Effects of egg yolk-citrate and milk extenders on chromatin structure and viability of cryopreserved bull sperm. Journal of Dairy Science, v.74, p.3836-48, 1991. KOZUMPL?K J., SOSNOV? J. The thermoresistance test of spermatozoa and fertility in bulls. Veterin?rn? Medic?na, v.30, p.385-92, 1985. KUMAR, S.; MILLAR, J.D.; WATSON, P.F. The effect of cooling rate on the survival of cryopreserved bull, ram, and boar spermatozoa: a comparison of two controlled-rate cooling machines. Cryobiology, v.46, p.24-53, 2003. 51 LANDA, C. A.; ALMQUIST, J. O. Effect of freezing large numbers of straws of bovine spermatozoa in an automatic freezer on post-thaw motility and acrossomal retention. Journal of Animal Science, v. 49, n. 5, p. 1190-1194, 1979. LIMA, V.H. Como e quando a sexagem de espermatoz?ides faz a diferen?a. DBO Gen?tica ? A revista de neg?cio da pecu?ria. p. 38-40, Setembro, 2005. LIU, Z.; FOOTE, H.R.; BROCKETT, C.C. Survival of bull sperm frozen at different rates in media varying in osmolatity. Crybiology, v.37, p.219-230, 1998. LOOMIS, P. R., GRAHAM, J. K. Commercial semen freezing: Individual male variation in cryosurvival and the response of stallion sperm to customized freezing protocols. Animal Reproduction Science, v.105, p.119?128, 2008. LOVE, C. C. The sperm chromatin structure assay: A review of clinical applications. Animal Reproduction Science, v.89, p.39-45, 2005. MATTIOLI, M.; BARBONI, B.; LUCIDI, P.; SEREN, E. Identification of capacitation in boar spermatozoa by chortetracicline staining. Theriogenology, v.45, p.373-381, 1996. MAXWELL, W.M.; EVANS, G.; HOLLINSHEAD, F.K.; BATHGATE, R.; DE GRAAF, S.P.; ERIKSSON, B.M.; et al. Integration of sperm sexing technology into the ART toolbox. Animal Reproduction Science. v.82-83, p.79?95, 2004. MAXWELL, W. M. C.; WELCH, G. R.; JOHNSON, L. A. Viability and membrane integrity of spermatozoa after dilution and flow cytometric sorting in the presence or absence of seminal plasma. Reproduction, Fertility and Development, v.8, p.1165-1178, 1997. MAZUR, P. Fundamental aspects of the freezing of cells, with emphasis on mammalian ova and embryos. In: International Congress on Animal Reproduction and AI. v.2, 99-114, 1980. MAZUR, P. Basic concepts in freezing cells. In: International Conference on Deep Freezing of Boar Semen. Uppsala, p. 25-27, 1985. 52 MEDEIROS, A.S.L., CARMO, M.T., GOMES, G.M.; et al. Criopreservation of stallion sperm using different amides. Theriogenology. V.58, p.273-276, 2002. MEIRELLES, C.; FARIA, V.R.; SOUZA, A.B.; WEISS, R.R.; SEGUI, M.S.; KOZICKI, L.E. Efici?ncia da insemina??o artificial com s?men sexado bovino: Aspectos de viabilidade reprodutiva e econ?mica. Archives of Veterinary Science, v.13, n.2, p.98-103, 2008. MENDOZA, C.; CARRERAS, A.; MOOS, J.; TESARIK, J. Distinction between true acrossome reaction and degenerative acrosome loss by a one-step staining method using Pisum sativum agglutinin. Journal of Reproduction and Fertility, v.95, p.755-763, 1992. MENEZES, E. S. B.; CAVALCANTE, J. M. M.; BRASIL, O. O.; SOUZA, D. F. R.; ASSIS NETO, E. T.; SILVA J?NIOR, J. B.; SALGUEIRO, C. C. M. ; NUNES, J. F. Uso da Nacetilciste?na na conserva??o de s?men ovino a 4oC: resultados preliminares. In: 35? Congresso Brasileiro de Medicina Veterin?ria, 2008, Gramado-RS. Anais do 35? Congresso Brasileiro de Medicina Veterin?ria, 2008. MIES FILHO, A. Insemina??o Articicial. Editora Sulina. 6? ed., v. 2, p. 750, Porto Alegre, 1987. MIRAGLIA, F.; MORAIS, Z. M.; CORTEZ, A.; MELVILLE, P. A.; MARVULLO, M. F. V.; RICHTZENHAIN, L. J.; VISINTIN, J. A.; VASCONCELLOS, S. A. Comparison of four antibiotics for inactivating leptospires in bull semen diluted in egg yolk extender and experimentally inoculated with Leptospira santarosai serovar Guaricura. Brazilian Journal of Microbiology, v. 34, n. 2, p. 147-151, 2003. MOC?, E.; GRAHAM, J.K.; SCHENK, J.L. Effect of sex-sorting on the ability of fresh and cryopreserved bull sperm to undergo an acrossome reaction. Theriogenology, v.66, p.929- 936, 2006. NAGASE, H. Scientific and technological Backgroung on pelet freezing of bull semen. Riproduzione Animale e Fecodazione Artificiale. Edagricole, p. 189-205, 1972. 53 NASH, T. Chemical constitution and physical properties of compounds able to protect living cells against damage due to freezing and thawing. In: MERYMAN, H.T., Cryobiology, New York: Academic Press. P.179-210, 1966. NIMBULKAR, M.V.; CHAUHAN, V.P.; HONMONDE, J. Preservation of goat semen at ambient temperatures.. In: International Conference on Goats, 1982. Tuckson. Proceedings. Tuckson: Dairy Goat Journal Publication, p.285, 1982. OLIVEIRA, K. M. Efeito do processo de sexagem de s?men por citometria de fluxo e criopreserva??o sobre a morfometria da cabe?a de espermatoz?ides e estabilidade de cromatina. 2007. 58f. Tese (mestrado). Faculdade de Medicina veterin?ria, Universidade Federal de Uberl?ndia, Uberl?ndia, 2007. PAPA, F.O., ZAHN, F.S., DELL?AQUA Jr., et al. Utiliza??o do diluente MP50 para criopreserva??o de s?men eq?ino. Revista Brasileira de Reprodrodu??o Animal. V. 26, p. 184-191, 2002. PAPAIOANNOU K. Z.; MURPHY R. P.; MONKS R. S.; HYNES N.; RYAN M. P.; BOLAND M.P.; ROCHE J. F. Assessment of viability and mitochondrial function of equine spermatozoa using double staining and flow cytometry. Theriogenology, v.48, p.299-312, 1997. PARKS, J. E.; LYNCH, D. V. Lipid composition and thermotrofic phase behavior of boar, bull, stallion and rooster sperm membrane. Cryobiology, v. 29, n. 2, p. 255-266, 1992. PE?A, A. L.; JOHANNISSON, A.; LINDE-FORSBERG, C. Post-thaw evaluation of dog spermatozoa using new triple fluorescent staining and flow cytometry. Theriogenology, v.52, p.965-980, 1999. PE?A, A. L.; QUINTELA, L. A.; HERRAD?N, P. G. Viability assessment of dog spermatozoa using flow cytometry. Theriogenology, v.50, p.1211-1220, 1998. 54 PICKETT, B.W.; SQUIRES, E.L.; MACKINNON, A. O. Procedures for collection, evaluation and utilization of stallion semen for artificial insemination. Fort Collins, CO: Colorado State University, Animal Reproduction Laboratory, 1987. POLGE, C.; SMITH, A. U.; PARKERS, A. S. Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature (London), v.164,p.666, 1949. ROYERE, D.; BARTHELEMY, C.; HAMAMAH, S.; LANSAC J. Cryopreservation of spermatozoa: a 1996 review. Human Reproduction Update, v. 2, n. 6 p. 553?559, 1996. SAHNI, K.L. Practical aspect of artificial insemination of goats in India. In: International Conference on Goats, 4, 1987. Bras?lia. Proceedings. Bras?lia: EMBRAPA-DTC, p.549-569, 1987. SALAMON, S; MAXWELL, W.M.C. Frozen storage of ram semen II. Causes of low fertility after cervical insemination and methods of improvement. Animal Reproduction Science, v. 38, p. 1-36, 1995. SIQUEIRA, J. B., GUIMAR?ES, J. D., COSTA, E. P., HENRY, M., TORRES, C. A. A., SILVA, M. V. G. B., SILVEIRA, T. S. Rela??o da taxa de gesta??o com s?men bovino congelado e testes de avalia??o esperm?tica in vitro. Revista Brasileira de Zootecnia, v.36, n.2, p.387-395, 2007. SOUZA, N. L. Avalia??o de t?cnicas para determinar a viabilidade e a integridade do acrossomo de espermatoz?ides criopreservados eq?inos. 2001. 76p. Disserta??o (Mestrado) ? Faculdade de Medicina Veterin?ria e Zootecnia, Universidade de S?o Paulo, Pirassununga, 2001. SUAREZ, S.S. The oviductal sperm reservoir mammals; mechanisms of formation. Biology of Reproduction, v.58, p.1105-1107, 1998. SZELL, S.; SHELTON, J.N. Role of equilibration before rapid freezing of mouse embryos. Journal of Reproduction and Fertility, v.78, p.699-703, 1986. 55 TESARIK, J.; MENDOZA, C, CARRERAS, A. Fast acrosome reaction measure: a highly sensitive method for evaluating stimulus-induced acrosome reaction. Fertility and Sterility, v.9, p.424-430, 1993. THOMAS, C. A.; GANER, D. L.; DEJARNETE, J. M.; MARSHALL, C. E. Effect of cryopreservation on bovine sperm organelle function and viability as determinated by flow cytometry. Biology of Reproduction, v.58, p.786-793, 1998. THUN, R.; HURTADO, M; JANETT, F. Comparison of Biociphos-Plus and TRIS-egg yolk extender for cryopreservation of bull semen.Theriogenology, v.57, p.1087-94, 2002. VALC?RCEL, A.; DE LAS HERAS, M. A.; P?REZ, L.; MOSES, D.F.; BALDASSARRE, H. Fluorescent staining as a method of assessing membrane damage and post-thaw survival of ram spermatozoa. Theriogenology, v.41, p.483-89, 1994. VALCARCEL, A., HERAS, M.A., MOSES, D.F. et al. Comparison between Sephadex G-10 and percoll for preparation of normospermic, asthenospermic and frozen/thawed ran semen. Animal Reproduction Science., v.41, p.215-224, 1996. VIANA, A. K. D. S. Avalia??o in vitro do s?men caprino resfriado a 4?C em dois m?todos e dois tipos de diluidores. 2003. 34p. Disserta??o (Mestrado em Medicina Veterin?ria Tropical) - Escola de Medicina Veterin?ria da Universidade Federal da Bahia, Salvador, 2003. VISHWANATH, R.; SHANNON, P. Storage of bovine semen in liquid and frozen state. Animal Reproduction Science, v. 62, p. 23-53, 2000. WATSON, P.F. The preservation of s?men in mammals. In: FINN, C.A. Oxford rewiews of reproductive biology. New York: Oxford University Press. 1, 283-330, 1979. WATSON, P.F. Recent development and concepts in the cryopreservation of spermatozoa and the assessment of their post-thawing function. Reproduction, Fertility and Development., v. 7, p.871-891, 1995. 56 WATSON , P.F. Cooling of spermatozoa and fertility capacity. Reproduction in Domestic Animals, v.31, p. 135-140,1999. WATSON, P. F. The causes of reduced fertility with cryopreserved semen. Animal Reproduction Science, v. 60-1, p. 481-492, 2000. WOELDERS, H., MATHIIJS, A., ENGEL, B. Effects of trehalose, and sucrose, osmalality of the freezing medium, and cooling rate on viability and intractness of sperm after freezing and thawing. Cryobiology, v.35, p.93-105, 1997.

Page generated in 0.0054 seconds