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Sperm quality gradually deteriorates with declines in motility and viability because an excess of reactive oxygen species (ROS) occurred during cooling storage. The objective of this study was to determine the effects of adding melatonin (an enhancer of free radical scavenging) during storage on semen quality in terms of sperm motility, viability, and functional integrity of plasma membrane. Semen was extended in the absence (control) and presence (1 μM) of melatonin in the Beltsville Thawing Solution medium, and stored for 7 days. Semen quality was assessed every 12 hours (until day 7). The results demonstrated that the differences in semen quality between the presence and absence of melatonin were significant after the storage time had passed 72 hrs for sperm motility, and 96 hrs for sperm viability and functional integrity of sperms (P < 0.01). In conclusion, the quality of semen in the presence of melatonin improved as the incubation time increased when compared to that of the control. The presence of melatonin in the BTS medium could be used to preserve the quality of sperms, making them last longer while being stored in a refrigerator at 15 °C.