This research aimed to examine the impact of resveratrol and catalase on the motility, plasma membrane integrity, acrosomal membrane integrity, and mitochondrial activity of Ankara buck semen following freeze-thawed process. In this study, semen samples obtained from four mature bucks were divided into four groups: control (C), resveratrol 500 µM/ml (R), catalase 50 IU/ml (CAT), and resveratrol 500 µM/ml + catalase 50 IU/ml (CATR). After dilution with Tris/egg yolk extender, the semen samples were frozen in liquid nitrogen and then thawed for assessment. The CATR group gave the highest values across all evaluated parameters (motility, plasma membrane integrity, acrosomal membrane integrity, and mitochondrial activity) compared to the other groups (61 ± 1.0%, 72.6 ± 0.70%, 70.73 ± 0.67%, 60.9 ± 0.79%, respectively) (p<0.05). In conclusion, the combination of catalase and resveratrol significantly improved the quality of buck semen after freeze-thawed process, thereby contributing to enhanced reproductive outcomes and genetic preservation.