Successful fertilization of Varicorhinus macrolepis eggs with sperm subjected to two freeze-thaw cycles
2008
Ji, X.S. | Zhao, Y. | Chen, S.L. | Jiang, Y.L. | Wang, H. | Song, J.Y. | Ding, L. | Chen, H.J.
Although sperm from several fish species have been successfully cryopreserved, few studies have been done in small and/or endangered species. The aim of the present work was to develop a method of freezing and refreezing Varicorhinus macrolepis semen in 1.8mL cryovials. The effect of extenders and cryoprotectants on the motility of post-thaw sperm was examined. The motility of frozen-thawed sperm in extender D-15 was higher than that in MPRS and fish Ringer solution (P <0.05). Dimethyl sulfoxide (DMSO) and glycerol provided greater protection to sperm than methanol during freezing and thawing; the most effective concentration of DMSO and glycerol was 10%. The fertilization rate of frozen-thawed sperm was not significantly different from that of fresh sperm. Furthermore, mean (±S.D.) hatching rate did not differ significantly between frozen-thawed (82.7±12.4%) and fresh sperm (90.7±4.5%). Although frozen-thawed sperm that was immediately refrozen had 0% post-thaw motility, frozen semen that was refrozen after dilution with D-15 (containing DMSO at a ratio of 1:2) had post-thaw motility of 38.3±2.9%. Motility was lower for refrozen than for frozen sperm (P <0.05). Furthermore, fertilization and hatching rates of refrozen sperm were 42.9±6.7 and 34.1±10.5%, respectively, which were lower than that of fresh sperm (P <0.05).
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