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5 h after being stored at 5°C). The diluted semen was loaded into 0.25 mL mini straws, equilibrated, and frozen using a freezing machine. Sperm motility, viability, membranes, DNA integrity, and concentrations of malondialdehyde (MDA) and aspartate aminotransferase (AST) enzymes were evaluated after thawing. The results showed that there were no significant differences in sperm motility and DNA integrity between dilutions (p 0.05). However, sperm viability and membrane intactness of one-step dilutions were higher than those of three-step