Abstract
Summary Cryo-preservation of carp, Cyprinus carpio, sperm Deep-freezing trials of carp sperm were carried out by varying several factors such as the basic saline solution, the cryoprotectors added (glycerol, propanediol, ethylene glycol and DMSO), the media (Menezo-INRA B2, egg yolk, urea) and the deep-freezing and dilution rates. The success of deepfreezing was judged by the percentage of motile spermatozoa, intensity of motility, fertilizing ability and morphological integrity of the spermatozoa studied under the scanning electron microscope. DMSO was the best cryoprotector and the mineral composition of the dilution medium the least important factor, but there was noticeable improvement after organic compounds were added. The following mixture has been proposed: NaCl 100 mM + KC1100 mM, Tris 20 mM, pH 8: 37%, Menezo medium B2 INRA: 15%, urea 5%, DMSO: 10%, fresh sperm: 33%. Optimal deep-freezing rate was: 5°C/min from 2 to-7°C and 25°C/min from-7 to-70°C. In these conditions, about 70 to 80% of the spermatozoa were motile after thawing compared to fresh control sperm, but fertilizing ability was not more than 30 to 40% of that with fresh sperm. The percentage of spermatozoa considered intact was 66% after thawing as against 83% for fresh control sperm. The motility and fertilizing ability of deep-frozen sperm were significantly improved when the dilution rate at insemination was reduced from 1/100 to 1/2.
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