Effects of Extenders, Cryoprotectants and Freezing Rate on Post-Thaw Sperm Motility of Walking Catfish (Clarias macrocephalus)
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Abstract
This research was aimed to study the effects of extenders, cryoprotectants, and freezing rate on post-thaw sperm motility of walking catfish (Clarias microcephalus) based on the use of the controlled-rate programmable freezer compared with styrofoam box. In the first experiment, freshly collected semen was diluted in Calcium-free Hank’s balanced salt solution (Ca-F HBSS) containing six different cryoprotectants including Dimethyl sulfoxide (DMSO), methanol, glycerol, sucrose, ethylene glycol, and propylene glycol at the final concentrations of 5%, 10% and 15% and the freezing rates of -3, -5, and -8°C/min. The highest post-thaw sperm motility (p<0.05) was obtained from the treatment using 10% DMSO and a freezing rate of -8°C/min, compared with other treatments using various combinations of cryoprotectants and freezing rate. The second experiment was performed by diluting fresh semen in one of the three extenders: Ca-F HBSS, 0.9% NaCl, and Extender 7 containing 10% DMSO before freezing within the styrofoam box at 2, 4, and 6 cm above the surface of liquid nitrogen (LN2) for 5, 10, and 15 minutes. The use of Ca-F HBSS with 10% DMSO and freezing at 6 cm above LN2 surface for 10 minutes had the highest post-thaw sperm motility (p<0.05) compared with other treatments. These results suggested that the styrofoam box method had a potential for cryopreservation of C. macrocephalus sperm, which could be applied for aquaculture and conservation of other aquatic animals.
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