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Fe 2C /ascorbate, hydrogen peroxide (H 2 O 2 ), and hypoxanthine/xanthine oxidase (XOD) are commonly used for inducing oxidative stress on spermatozoa. A comparative study of these agents was carried out on thawed spermatozoa from red deer. First, we tested a high, medium, and low concentration of each agent: 100, 10, and 1 mM Fe 2C (hydroxyl radical generator); 1 mM, 100, and 10 mM H 2 O 2 ; and 100, 10, and 1 mU/ml XOD (superoxide and H 2 O 2 generator), incubated at 37 8C for 180 min. Intracellular reactive oxygen species (ROS; H 2 DCFDA) increased with dose and time similarly for the three systems at each concentration level. Motility and mitochondrial membrane potential (Dj m ) were considerably decreased by H 2 O 2 (1 mM and 100 mM) and XOD (100 and 10 mU/ml

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