Pretreatment of sperm for oxidative and antioxidative analysis has been shown to be different in studies. These differences can affect the result of analyses. The aim of this study was to compare the analyses of sperm homogenization methods. The semen of four rams were used in the study. Pretreatment procedures included the rotary, bead, and ultrasonic homogenization methods. Oxidative and antioxidative status of sperm samples was assessed by measuring serum lipid hydroperoxide, total oxidant status, free sulfhydryl groups (SH total thiol), ceruloplasmin, and total antioxidant capacity levels. As a result, the homogenization methods before oxidative and antioxidative status analyses significantly affected the result (P