Abstract: Rabbit sperm are known to undergo damage during both cryopreservation and thawing, leading
to decreased viability, motility and membrane integrity. Glycine can protect sperm and reduce damage
during freezing. Swim-up is a simple semen processing method for selecting good motile sperm. The
study evaluated the effect of the swim-up method and glycine with different concentrations supplemented
to the frozen medium. Three indigenous black rabbits were selected for semen collection by artificial
vagina. Next, semen was selected by swim-up method and diluted with glycine-added frozen medium.
The samples were then transferred to 0.5 mL straws, cooled to 15°C and 5°C, placed in liquid nitrogen
vapour, and finally placed directly into liquid nitrogen (-196°C). The samples were thawed and evaluated
for sperm quality. The results showed that the medium supplemented with 10mM glycine in combination
with swim-up method for 30 min gave the best results and was significantly different from the remaining
concentrations (P49.7%, respectively. In conclusion, 10 mM glycine concentration combined with swim-up for 30 min is
the optimal choice for freezing local black rabbit semen. The study highlights the importance of optimising
freezing protocols to improve the quality of frozen rabbit sperm, which can have important implications for
animal breeding and conservation efforts