Extenders and semen:extender ratio in the cryopreservation of sperm of fat snook Centropomus parallelus
Keywords:
fat snook, Centropomus parallelus, semen, cryopreservation, extender, floating net cageAbstract
Aiming to develop semen cryopreservation methods of the fat snook, Centropomus parallelus, studies were carried out in the Instituto de Pesca-SP, to optimize the sperm motility and motility time and semen fertilization capacity. Fifty reproducers, originated from artificial reproduction, were brought up in cages that were set in oceanic environment. The semen, diluted in three Ringer’s solutions (pH 6.1, 7.8 and 8.2) in the proportions of 1:1, 1:2, 1:3, 1:4 and 1:5, was frozen by nitrogen vapor and later on, transferred to liquid nitrogen. The cooling rate, the dimethyl sulfoxide (DMSO) cryoprotector concentration and the equilibration time were 90°C min-1, 10% and 60 seconds, respectively. The fertilization capacity was determined by calculating volumes of semen necessary to obtain the sperm:egg proportion of 4x104, 1x105, 1.6x105 and 2.2x105. Regarding the spawning eggs, the females received human chorionic gonadotropin (hCG) and 1 IU/g body weight. Regarding fresh semen, the motility was 100%, as the motility time was 464.6 ± 47.56 seconds. For cryopreserved semen, the best results for a 70.0 ± 3.94% motility and 610.8 ± 9.60 seconds motility time, using the diluent with an 8.2 pH. No significant differences (p<0.01) were observed regarding the different semen:diluent proportions. Finally, equal (p<0.01) fertilization rates (99.0 ± 0.66%) were obtained from 1.6x105 sperm:egg concentration, indicating that cryopreservation sets itself as an important resource for semen stocking and the fat snook reproduction control.
References
ALAVI, S.M.H.; COSSON, J.; KARAMI, M.;ABDOULHAY, H.; MOZAJI AMIRI, B. 2004
Chemical composition and osmolality of seminal plasma of Acipenser persicus, their
physiological relationsheap with sperm motility. Aquac. Res., 35: 1238-1243.
ALVAREZ-LAJONCHÉRE, L. y MOLEJÓN, O.G.H. 2001 Producción de juveniles de peces estuarinos para un centro en América Latina y el Caribe: diseí±o,operación y tecnologías. The World Aquaculture Society, Baton Rouge, LA. 424p.
BILLARD, R. 1978 Some data on gametes preservation and artificial insemination in teleost fish. Actes de colloques du CNEXO 8: 59-73.
BILLARD, R. 1990 Artificial insemination in fish. In:LAMMING, G.E. Marshall’s physiology of reproduction. Churchill Livingstone: Edinburg,2: 870-888.
BILLARD, R. and COSSON, J. 1988 Sperm motility in rainbow trout, Parasalmo gairdneri: effects of pH and temperature. In Breton B. and Zohar Y. eds. Reproduction in fish basic and applied aspects in endocrinology and genetics. Paris:
INRA. p. 161-167.
BILLARD, R.; COSSON, J.; CRIM, L.W. 1993 Motility of fresh and aged halibut sperm. Aquatic Living Resources, 6: 67-75.
BILLARD, R.; COSSON, J.; CRIM, L.M.; SUQUET, M.1996 Sperm physiology and quality. In:BROMAGE N.R. and ROBERTS, R.J. Broodstock management and egg and larval quality. Oxford:Blackwell Science. p. 25-52.
BLAXTER, J.H.S. 1953 Sperm storage and crossfertilization of spring and autumn spawning herring. Nature, 172: 1189-1190.
CHAO, N.H.; CHEN, H.P.; LIAO, I.C. 1975 Study on cryogenic preservation of grey mullet sperm. Aquaculture, 5: 389-406.
CHAO, N.H.; TSAI, H.P.; LIAO, I.C. 1992 Short and long-term cryopreservation of sperm and sperm suspension of the grouper, Epinephelus malabaricus (Bloch and Schneider). Asian Fishery Science, 5: 103-116.
CIERESZKO, A.; Glogowski, J.; Dabrowski, K. 2000 Biochemical characteristics of seminal plasma and spermatozoa. In: TIERSCH, T.R. and MAZIC, P.M. Cryopreservation in aquatic species.World Aquaculture Society, Baton Rouge,
Louisiana. p.20-48.
COSSON, J. 2004 The ionic and osmotic factors controlling motility of fish spermatozoa.Aquac. Int. 12: 69-85.
COSSON, J.; GROISON, A.-L.; SUQUET, M.; FAUVEL, C.;DREANNO, C.; BILLARD, R. 2008a Studing sperm motility in marine fish: an overview on the state of the art. J. Appl. Ichthyol. 24: 460-486.
COSSON, J.; GROISON, A.-L.; SUQUET, M.; FAUVEL, C.;DREANNO, C.; BILLARD, R. 2008b Marine fish spermatozoa: racing ephemeral swimmers.Reproduction. 136: 277-294.
DREANNO, C.; SUQUET, M.; QUEMENER, L.; COSSON, J.; FIERVILLE, F.; NORMANT, Y.; BILLARD, R.1997 Cryopreservation of turbot (Scophthalmus maximus) spermatozoa. Theriogenology, 48: 589-603.
FAUVEL, C.; SUQUET, M.; DREANNO, C.; ZONNO, V.; MENU, B. 1998 Cryopreservation of sea bass (Dicentrarchus labrax) spermatozoa in experimental and production simulating conditions. Aquat. Living Resour., 11: 387-394.
FAUVEL, C.; SAVOYE, O.; DREANNO, C.; COSSON, J.;SUQUET, M. 1999 Characteristics of captive seabass in relation to its fertilization potential.Journal of Fish Biology, 54: 356-369.
FERRAZ, E.M.; CERQUEIRA, V.R.; ALVAREZLAJONCHíˆRE, L.; CANDIDO, S. 2003. Indução í desova do robalo-peva, Centropomus parallelus, através de injeção e implante de LHRHa. Bol. Inst. Pesca, São Paulo, 28(2): 125-133.
GARZÓN, D.L.; PEí"˜ARANDA, D.S.; PÉREZ, L.;MARCO-JIMÉNEZ, F.; ESPERt, X.; MíÅ“LLER, T.; JOVER, M.; ASTURIANO, J.F. 2008 Effects of pH, sodium bicarbonate, cryoprotectants and foetal bovine serum on the cyopreservation of european eel sperm. Reprod. Dom Anim., 43:99-105.
GODINHO, H. M.; SERRALHEIRO, P. C. S.; FERRAZ,E.M.; PIMENTEL, C. M. M.; OLIVEIRA, I.R. e PAIVA, P. 2000 Reprodução induzida em robalo Centropomus parallelus Poey, 1860. Braz.J.Res.anim.Sci., São Paulo, 37(1): 37-42.
GWO, J.C. 2000 Cryopreservation of sperm of some marine fishes. In: TIERSCH, T.R. and MAZIC, P.M. Cryopreservation in aquatic species. World Aquaculture Society, Baton Rouge,Louisiana. p.138-160.
INABA, K, 2003 Molecular architecture of the sperm flagella: molecules for motility and signaling. Zool. Sci., 20: 1043-1056.
LEE, H.C.; HOHNSON C.; EPEL, D. 1983 Changes in internal pH associated with initiation of motility and acrosome reaction of sea urchin sperm. Dev. Biol., 95: 31-45.
LAHNSTEINER, F.; WEISMANN, T.; PATZNER, R.A. 1997 Metanol as cryoprotectant and the suitability of 1.2 mL and 5 mL straws for cryopreservation of semen from salmonid fishes. Aquaculture Reserch, Salzburg, 28: 471-479.
LEUNG, L. K. P. 1987 Cryopreservation of spermatozoa of the barramundi, Lates
calcarifer (Teleostei: Centropomidae).Aquaculture, Amsterdam, 64: 243-247.
LUBZENS, E.; DAUBE, N.; PEKARSDY, I.; MAGNUS,Y.;COHEN, A.; YUSEFOVICH, F.; FEIGIN, P. 1997 Carp (Cyprinus carpio L.) spermatozoa cryobanks: strategies in research and application. Aquaculture, Amsterdam, 155: 13-30.
Mí€RIí€N, T.; KRASZNAI, Z.; BALKAY, L.; EMRI, M.;TRÓN, L. 1997 Role of extracellular and intracellular pH in carp sperm motility and modifications by hyperosmosis of regulation of the Na+/H+ exchanger. Cytometry, 27: 374-382.
MC ANDREW, B.J.; RANA, K.J.; PENMAN, D.J. 1993 Conservation and preservation of genetic variation in aquatic organism. In: MUIR, J.F. and ROBERTS, R.J. Recent Advances in: Aquaculture, vol IV, Oxford, Blackwell. p.295-336.
MEDINA, V.; VELASCO, Y.; CRUZ, P. 2005 Aspectos generales de la criopreservation espermática en peces teleósteos. Rev. Colomb. Cienc.Pecuar., 18: 34-48.
MIOSO, R. 1995 Indução í reprodução e incubação de ovos de robalo Centropomus parallelus Poey, 1860 (Pisces, Centropomidae). Florianópolis. 51p.(Dissertação de Mestrado. Universidade Federal de Santa Catarina).
MOUNIB, M.S.; HWANG, P.C.; IDLER, D.R. 1968 Cryogenic preservation of Atlantic cod Gadus morhua sperm. Journal of the Fisheries,Canada, 25: 2623-2632.
ODA, S. and MORISAWA, M. 1993a Rises of intracellular Ca+ and pH mediate the
initiation of sperm motility in marine teleosts. Cell Motil. Cytoskeleton. 25: 171-178.
ODA, S. and MORISAWA, M. 1993b Rises of intracellular Ca+ and pH mediate the
initiation of sperm of farmed european eel Anguilla Anguilla. J. World Aquac. Soc. 35:
240-246.
PELETEIRO, J.B.; CHEREGUINI, O.; CAL, R.M. 1996 Preliminary results of artificial fertilization carried out with cryopreserved sperm of turbot Scophthalmus maximus (Linnaeus, 1758). Informe Tecnico del Instituto Espanological de Oceanografo, 162: 1-13.
RANA, K.J.; GUPTA, S.D.; MCANDREW, B.J. 1992 The relevance of collection techniques on the quality of manually stripped Atlantic salmon (Salmo salar) milt. In: WORKSHOP ON GAMETE AND EMBRYO STORAGE AND CRYOPRESERVATION IN AQUATIC ORGANISMS. 30 march í 2 april/1992.
RANA, K.J. 1996 Preservation of gametes. In:BROMAGE, N.R. and ROBERTS,R.J. Broodstock management and egg and larval quality.Oxford: Blackwell Science, p.53-75.
RIDEOUT, R.M.; TRIPPEL, E.A.; LITVAK, M.K. 2004 The development of haddock and Atlantic cod sperm cryopreservation techniques and effect of sperm age on cryopreservation success. Journal of Fish Biology, St. John’s, 65:299-311.
RILEY, K.L.; HOLLADAY, C.G.; CHESNEY, E.J.;TIERSCH, T.R. 2004 Cryopreservation of sperm of red snapper. Aquaculture, 238: 183-194.
SANCHES, E.G.; OLIVEIRA, I.R.; SERRALHEIRO, P.C.S.2008 Crioconservação do sêmen da garoupaverdadeira Epinephelus marginatus. Bioikos, 22 (2): 81-90.
SANSONE, G.; FABRIOCINI, A.; IEROPOLI, S.;LANGELLOTTI, A.L.; OCCIDENTE, M.;
MATASSINO, D. 2002 Effects of extender composition, cooling rate, and freezing on the motility of sea bass (Dicentrarchus labrax, L.) spermatozoa after thawing. Cryobiology,Elsevier Science, Napoli, 44: 229-239.
SCOTT, A.P. and BAYNES, S.M. 1980 A review of the biology, handling and storage of salmonid spermatozoa. J. Fish Biol., 17: 707-739.
SERRALHEIRO, P.C.S.; OLIVEIRA, I.R.; GODINHO,H.M. 1998 Fertilização de ovócitos de robalo Centropomus parallelus, com sêmen crioconservado. In: SIMPÓSIO BRASILEIRO DE AQíÅ“ICULTURA, 12, Recife, 1998. Anais…Recife, p.360-1.
SERRALHEIRO, P.C. DA S.; FOGLI DA SILVEIRA,W.;GODINHO, H.M.; OLIVEIRA, I DA R. 1999. O uso de três soluções diluidoras em sêmen de tainha í Mugil platanus, Günther, 1880,resfriado em container de vapor de nitrogênio líquido. In: ENCONTRO BRASILEIRO DE ICTIOLOGIA, 13, São Carlos. Anais... p.508.
STOSS, J. 1983 Fish gamete preservation and spermatozoan physiology. In: HOAR, W.S.; RANDALL, D.J.; DONALDSON, E.M. (Eds.). Fish physiology. New York: Academic Press.p.305-350.
SUQUET, M. 1992 La production de sperme chez le turbot, aspects descriptifs et experimentaux. Mémorie EPHE, Paris.
SUQUET, M.; DREANNO, C.; FAUVEL, C.; COSSON,J.; BILLARD, R. 2000 Cryopreservation ofsperm in marine fish. Aquaculture Research,31: 231-243.
TANAKA, S.; ZANG, H.; RORIE, N.; YAMADA, Y.; OKAMURA, A.; UTOH, T., MIKAWA, N.; OKA, H.P.; KUROKURA, H. 2002 Long-term cryopreservation of sperm of Japanese eel. J.Fish Biol. 60: 139-146.
TIERSCH, T.R. 2000 Introduction. In: TIERCH, T.R.and MAZUR P.M. Cryopreservation in Aquatic Species. World Aquaculture Society. Baton Rouge, Louisiana. p.xix-xxvi.
TIERSCH, T.R.; WAYMAN, W.R.; SKAPURA, D.P.;NEIDIG, C.L.; GRIER, H.J. 2004 Transport and cryopreservation of sperm of the common snook, Centropomus undecimalis (Bloch). Aquaculture Research. Baton Rouge, Louisiana,35: 278-288.
ZILLI, L.; SCHIAVONE, R.; ZONNO, V.; ROSSANO, R.;STORELLI, C.; VILELLA, S. 2005 Effect of cryopreservation on sea bass sperm proteins.Biology of Reproduction, Lecce, 72: 1262-1267.
Chemical composition and osmolality of seminal plasma of Acipenser persicus, their
physiological relationsheap with sperm motility. Aquac. Res., 35: 1238-1243.
ALVAREZ-LAJONCHÉRE, L. y MOLEJÓN, O.G.H. 2001 Producción de juveniles de peces estuarinos para un centro en América Latina y el Caribe: diseí±o,operación y tecnologías. The World Aquaculture Society, Baton Rouge, LA. 424p.
BILLARD, R. 1978 Some data on gametes preservation and artificial insemination in teleost fish. Actes de colloques du CNEXO 8: 59-73.
BILLARD, R. 1990 Artificial insemination in fish. In:LAMMING, G.E. Marshall’s physiology of reproduction. Churchill Livingstone: Edinburg,2: 870-888.
BILLARD, R. and COSSON, J. 1988 Sperm motility in rainbow trout, Parasalmo gairdneri: effects of pH and temperature. In Breton B. and Zohar Y. eds. Reproduction in fish basic and applied aspects in endocrinology and genetics. Paris:
INRA. p. 161-167.
BILLARD, R.; COSSON, J.; CRIM, L.W. 1993 Motility of fresh and aged halibut sperm. Aquatic Living Resources, 6: 67-75.
BILLARD, R.; COSSON, J.; CRIM, L.M.; SUQUET, M.1996 Sperm physiology and quality. In:BROMAGE N.R. and ROBERTS, R.J. Broodstock management and egg and larval quality. Oxford:Blackwell Science. p. 25-52.
BLAXTER, J.H.S. 1953 Sperm storage and crossfertilization of spring and autumn spawning herring. Nature, 172: 1189-1190.
CHAO, N.H.; CHEN, H.P.; LIAO, I.C. 1975 Study on cryogenic preservation of grey mullet sperm. Aquaculture, 5: 389-406.
CHAO, N.H.; TSAI, H.P.; LIAO, I.C. 1992 Short and long-term cryopreservation of sperm and sperm suspension of the grouper, Epinephelus malabaricus (Bloch and Schneider). Asian Fishery Science, 5: 103-116.
CIERESZKO, A.; Glogowski, J.; Dabrowski, K. 2000 Biochemical characteristics of seminal plasma and spermatozoa. In: TIERSCH, T.R. and MAZIC, P.M. Cryopreservation in aquatic species.World Aquaculture Society, Baton Rouge,
Louisiana. p.20-48.
COSSON, J. 2004 The ionic and osmotic factors controlling motility of fish spermatozoa.Aquac. Int. 12: 69-85.
COSSON, J.; GROISON, A.-L.; SUQUET, M.; FAUVEL, C.;DREANNO, C.; BILLARD, R. 2008a Studing sperm motility in marine fish: an overview on the state of the art. J. Appl. Ichthyol. 24: 460-486.
COSSON, J.; GROISON, A.-L.; SUQUET, M.; FAUVEL, C.;DREANNO, C.; BILLARD, R. 2008b Marine fish spermatozoa: racing ephemeral swimmers.Reproduction. 136: 277-294.
DREANNO, C.; SUQUET, M.; QUEMENER, L.; COSSON, J.; FIERVILLE, F.; NORMANT, Y.; BILLARD, R.1997 Cryopreservation of turbot (Scophthalmus maximus) spermatozoa. Theriogenology, 48: 589-603.
FAUVEL, C.; SUQUET, M.; DREANNO, C.; ZONNO, V.; MENU, B. 1998 Cryopreservation of sea bass (Dicentrarchus labrax) spermatozoa in experimental and production simulating conditions. Aquat. Living Resour., 11: 387-394.
FAUVEL, C.; SAVOYE, O.; DREANNO, C.; COSSON, J.;SUQUET, M. 1999 Characteristics of captive seabass in relation to its fertilization potential.Journal of Fish Biology, 54: 356-369.
FERRAZ, E.M.; CERQUEIRA, V.R.; ALVAREZLAJONCHíˆRE, L.; CANDIDO, S. 2003. Indução í desova do robalo-peva, Centropomus parallelus, através de injeção e implante de LHRHa. Bol. Inst. Pesca, São Paulo, 28(2): 125-133.
GARZÓN, D.L.; PEí"˜ARANDA, D.S.; PÉREZ, L.;MARCO-JIMÉNEZ, F.; ESPERt, X.; MíÅ“LLER, T.; JOVER, M.; ASTURIANO, J.F. 2008 Effects of pH, sodium bicarbonate, cryoprotectants and foetal bovine serum on the cyopreservation of european eel sperm. Reprod. Dom Anim., 43:99-105.
GODINHO, H. M.; SERRALHEIRO, P. C. S.; FERRAZ,E.M.; PIMENTEL, C. M. M.; OLIVEIRA, I.R. e PAIVA, P. 2000 Reprodução induzida em robalo Centropomus parallelus Poey, 1860. Braz.J.Res.anim.Sci., São Paulo, 37(1): 37-42.
GWO, J.C. 2000 Cryopreservation of sperm of some marine fishes. In: TIERSCH, T.R. and MAZIC, P.M. Cryopreservation in aquatic species. World Aquaculture Society, Baton Rouge,Louisiana. p.138-160.
INABA, K, 2003 Molecular architecture of the sperm flagella: molecules for motility and signaling. Zool. Sci., 20: 1043-1056.
LEE, H.C.; HOHNSON C.; EPEL, D. 1983 Changes in internal pH associated with initiation of motility and acrosome reaction of sea urchin sperm. Dev. Biol., 95: 31-45.
LAHNSTEINER, F.; WEISMANN, T.; PATZNER, R.A. 1997 Metanol as cryoprotectant and the suitability of 1.2 mL and 5 mL straws for cryopreservation of semen from salmonid fishes. Aquaculture Reserch, Salzburg, 28: 471-479.
LEUNG, L. K. P. 1987 Cryopreservation of spermatozoa of the barramundi, Lates
calcarifer (Teleostei: Centropomidae).Aquaculture, Amsterdam, 64: 243-247.
LUBZENS, E.; DAUBE, N.; PEKARSDY, I.; MAGNUS,Y.;COHEN, A.; YUSEFOVICH, F.; FEIGIN, P. 1997 Carp (Cyprinus carpio L.) spermatozoa cryobanks: strategies in research and application. Aquaculture, Amsterdam, 155: 13-30.
Mí€RIí€N, T.; KRASZNAI, Z.; BALKAY, L.; EMRI, M.;TRÓN, L. 1997 Role of extracellular and intracellular pH in carp sperm motility and modifications by hyperosmosis of regulation of the Na+/H+ exchanger. Cytometry, 27: 374-382.
MC ANDREW, B.J.; RANA, K.J.; PENMAN, D.J. 1993 Conservation and preservation of genetic variation in aquatic organism. In: MUIR, J.F. and ROBERTS, R.J. Recent Advances in: Aquaculture, vol IV, Oxford, Blackwell. p.295-336.
MEDINA, V.; VELASCO, Y.; CRUZ, P. 2005 Aspectos generales de la criopreservation espermática en peces teleósteos. Rev. Colomb. Cienc.Pecuar., 18: 34-48.
MIOSO, R. 1995 Indução í reprodução e incubação de ovos de robalo Centropomus parallelus Poey, 1860 (Pisces, Centropomidae). Florianópolis. 51p.(Dissertação de Mestrado. Universidade Federal de Santa Catarina).
MOUNIB, M.S.; HWANG, P.C.; IDLER, D.R. 1968 Cryogenic preservation of Atlantic cod Gadus morhua sperm. Journal of the Fisheries,Canada, 25: 2623-2632.
ODA, S. and MORISAWA, M. 1993a Rises of intracellular Ca+ and pH mediate the
initiation of sperm motility in marine teleosts. Cell Motil. Cytoskeleton. 25: 171-178.
ODA, S. and MORISAWA, M. 1993b Rises of intracellular Ca+ and pH mediate the
initiation of sperm of farmed european eel Anguilla Anguilla. J. World Aquac. Soc. 35:
240-246.
PELETEIRO, J.B.; CHEREGUINI, O.; CAL, R.M. 1996 Preliminary results of artificial fertilization carried out with cryopreserved sperm of turbot Scophthalmus maximus (Linnaeus, 1758). Informe Tecnico del Instituto Espanological de Oceanografo, 162: 1-13.
RANA, K.J.; GUPTA, S.D.; MCANDREW, B.J. 1992 The relevance of collection techniques on the quality of manually stripped Atlantic salmon (Salmo salar) milt. In: WORKSHOP ON GAMETE AND EMBRYO STORAGE AND CRYOPRESERVATION IN AQUATIC ORGANISMS. 30 march í 2 april/1992.
RANA, K.J. 1996 Preservation of gametes. In:BROMAGE, N.R. and ROBERTS,R.J. Broodstock management and egg and larval quality.Oxford: Blackwell Science, p.53-75.
RIDEOUT, R.M.; TRIPPEL, E.A.; LITVAK, M.K. 2004 The development of haddock and Atlantic cod sperm cryopreservation techniques and effect of sperm age on cryopreservation success. Journal of Fish Biology, St. John’s, 65:299-311.
RILEY, K.L.; HOLLADAY, C.G.; CHESNEY, E.J.;TIERSCH, T.R. 2004 Cryopreservation of sperm of red snapper. Aquaculture, 238: 183-194.
SANCHES, E.G.; OLIVEIRA, I.R.; SERRALHEIRO, P.C.S.2008 Crioconservação do sêmen da garoupaverdadeira Epinephelus marginatus. Bioikos, 22 (2): 81-90.
SANSONE, G.; FABRIOCINI, A.; IEROPOLI, S.;LANGELLOTTI, A.L.; OCCIDENTE, M.;
MATASSINO, D. 2002 Effects of extender composition, cooling rate, and freezing on the motility of sea bass (Dicentrarchus labrax, L.) spermatozoa after thawing. Cryobiology,Elsevier Science, Napoli, 44: 229-239.
SCOTT, A.P. and BAYNES, S.M. 1980 A review of the biology, handling and storage of salmonid spermatozoa. J. Fish Biol., 17: 707-739.
SERRALHEIRO, P.C.S.; OLIVEIRA, I.R.; GODINHO,H.M. 1998 Fertilização de ovócitos de robalo Centropomus parallelus, com sêmen crioconservado. In: SIMPÓSIO BRASILEIRO DE AQíÅ“ICULTURA, 12, Recife, 1998. Anais…Recife, p.360-1.
SERRALHEIRO, P.C. DA S.; FOGLI DA SILVEIRA,W.;GODINHO, H.M.; OLIVEIRA, I DA R. 1999. O uso de três soluções diluidoras em sêmen de tainha í Mugil platanus, Günther, 1880,resfriado em container de vapor de nitrogênio líquido. In: ENCONTRO BRASILEIRO DE ICTIOLOGIA, 13, São Carlos. Anais... p.508.
STOSS, J. 1983 Fish gamete preservation and spermatozoan physiology. In: HOAR, W.S.; RANDALL, D.J.; DONALDSON, E.M. (Eds.). Fish physiology. New York: Academic Press.p.305-350.
SUQUET, M. 1992 La production de sperme chez le turbot, aspects descriptifs et experimentaux. Mémorie EPHE, Paris.
SUQUET, M.; DREANNO, C.; FAUVEL, C.; COSSON,J.; BILLARD, R. 2000 Cryopreservation ofsperm in marine fish. Aquaculture Research,31: 231-243.
TANAKA, S.; ZANG, H.; RORIE, N.; YAMADA, Y.; OKAMURA, A.; UTOH, T., MIKAWA, N.; OKA, H.P.; KUROKURA, H. 2002 Long-term cryopreservation of sperm of Japanese eel. J.Fish Biol. 60: 139-146.
TIERSCH, T.R. 2000 Introduction. In: TIERCH, T.R.and MAZUR P.M. Cryopreservation in Aquatic Species. World Aquaculture Society. Baton Rouge, Louisiana. p.xix-xxvi.
TIERSCH, T.R.; WAYMAN, W.R.; SKAPURA, D.P.;NEIDIG, C.L.; GRIER, H.J. 2004 Transport and cryopreservation of sperm of the common snook, Centropomus undecimalis (Bloch). Aquaculture Research. Baton Rouge, Louisiana,35: 278-288.
ZILLI, L.; SCHIAVONE, R.; ZONNO, V.; ROSSANO, R.;STORELLI, C.; VILELLA, S. 2005 Effect of cryopreservation on sea bass sperm proteins.Biology of Reproduction, Lecce, 72: 1262-1267.
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