CRYOPRESERVATION OF RAINBOW TROUT SEMEN: DILUENT, STRAW AND THE VAPOR COLUMN
Keywords:
fish, spermatozoa semen, cryopreservation, rainbow trout, Oncorhynchus mykissAbstract
The present paper concerns to the development of a simple semen freezing technique to be used in commercial systems. Fresh milt was obtained from two years-old adult males of rainbow trout. The samples were previously cooled (4°C), then diluted, packaged in straws, and frozen in liquid nitrogen vapor. The freezing system was composed by a 33 L StyrofoamTM box with a graduated metal web; straws of 0.5 mL and 4 mL were tested and two types of extenders were used: V2e with 10% DMSO and modified Cortland solution with 24% DMSO. Two experiments were performed: the first one evaluated the effect of the extenders and straw type on the fertility of the frozen/thawed semen, and the second one verified the effect of the vapor column and straw position on the viability of the frozen/thawed semen, using 0.5 mL-straws and V2e extender. The sperm viability was tested by checking eyed eggs production. In the first experiment, there were no statistical differences (P>0.05) between the two tested straws and the modified Cortland solution. However, the V2e solution treatment showed a significantly higher fertilization rate when associated to 0.5 mL straw. In the second experiment, a remarkable statistical interaction (P>0.05) was observed between the straw cargo and vapor column (freezing point) effects on the percentage of eyed eggs.
References
CLOUD, J.G.; MILLER, W.H.; LEVANDUSKI, M.J. 1990 Cryopreservation of sperm as a means to store salmonid germ plasm and to transfer genes from wild fish to hatchery populations. The Progressive Fish Culturist, 52: 51-53
CONGET, P.; FERNANDEZ, M.; HERRERA, G.; MINGUELL. J.J. 1996 Cryopreservation of rainbow trout (Oncorhynchus mykiss) spermatozoa using programmable freezing. Aquaculture, Santiago de Chile,143: 319-329.
JAMIENSON, B.G.M. 1991 Fish evolution and systematic: evidence from spermatozoa. Cambridge:University Press 319p.
FOGLI DA SILVEIRA, W.; KAVAMOTO, E.T.; RIGOLINO, M.G.; ARRUDA SOARES, H.; SILVEIRA, A.N.; VERÍSSIMO, R. 1994 Congelação do sêmen de truta arco-íris, Oncorhynchus mykiss, em vapor de nitrogênio líquido. B. Inst. Pesca, São Paulo, 21(único): 55-60.
HOLTZ, W. 1993 Cryopreservation of rainbow-trout (Oncorhynchus mykiss) sperm í practical recommendations. Aquaculture, Amsterdam, 110(1): 97-100.
LAHNSTEINER, F.; WEISMANN, T.; PATZNER, A.R. 1997 Methanol as cryoprotectant and the suitability of 1.2 ml and 5 ml straws for cryopreservation of semen from salmonid fishes. Aquaculture Research,28: 471-479.
RICHARDSON, G.F.; WILSON, C.E.; CRIM, L.W.; YAO, Z. 1999 Cryopreservation of yellowtail flounder (Pleuronectes platessa) semen in large straws. Aquaculture, 174: 89-94.
SALISBURY, G.W. and VANDEMARK, N.L. 1964 Fisiología de la reproducción e inseninación artificial de los bovidos. Trad. D. José Maria Santiago Luque.Zaragosa: Acribia. 707p.
STEINBERG, H.; HEDDER, A.; BAULAIN, R.; HOLTZ, W. 1995 Cryopreservation of rainbow trout, Onchorhyncus mykiss semen straws. In: INTERNATIONAL SYMPOSIUM ON THE REPRODUCTIVE PHYSIOLOGY OF FISH, 5,Austin, 1995. Proceedings… Austin, University of Texas, USA. p.146.
STEIN, H. and BAYRLE, H. 1978 Cryopreservation of the sperm of some freshwater teleosts. Ann. Biol. Anim. Bioch. Biophys., Paris, 18(4): 1073-1076.
WEISMANN, T.; LAHNSTEINER, F.; PATZNER, R.A. 1995 Cryopreservation of semen of salmonid fishes and its adaptation for practical application. In: INTERNATIONALSYMPOSIUM ON THE REPRODUCTIVE PHYSIOLOGY OF FISH, 5, Austin, 1995. Proceedings… Austin, University of Texas,USA. p.90-92.
WHEELER, P.A. and THORGAARD, G.H. 1991 Cryopreservation of rainbow trout semen in large straws. Aquaculture, 93(1): 95-100.