The Honey Solution Effect as A Natural Cryoprotectant on Sperm Quality of Botia, Chromobotia Macracanthus Bleeker 1852

The purpose of study was to obtain the best combination of natural cryoprotectant (honey solution) and synthetic cryoprotectant (methanol) on the quality of spermatozoa Chromobotia macracanthus Bleeker 1852, after freezing at -80 ℃ for 48 hours. The combination of 10% methanol with various concentrations of honey solution (0%, 0.1%, 0.3%, 0.5%, 0.7% and 0.9%) was tested in this study. Ringer's solution was used as an extender. The diluted sperm was then equilibrated for 25 minutes at 4 ℃, then frozen at -80 ℃ for 48 hours. The sperm was then thawed at 40 ℃ for 13 seconds. Viability, motility and percentage of fertilization were evaluated. The results of the analysis of variance showed that the combination of 10% methanol with several concentrations of honey solution had a significant effect (P <0.05) on the viability of spermatozoa and the percentage of fertilization, but had no effect on sperm motility (P> 0.05). The combination of 0.1% honey solution and 10% methanol showed the highest percentage of both motility (89.4 ± 5.45%), viability (85.75 ± 4.79%), and the percentage of fertilization (98.55 ± 1.69%). %). The conclusion of this study is that 0.1% honey solution combined with 10% methanol in Ringer's solution is the best cryoprotectant for C. macracanthus spermatozoa stored at -80 ℃ for 48 hour