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This study evaluates the effects of pumpkin seed oil and sunflower oil along with vitamin E on the reproductive characteristics of aged roosters. Thirty Ross breeder roosters (45-wk-old) were assigned into 6 groups (5 birds/group) with the following diets: 1) control group (basal diet), 2) basal diet with 2% pumpkin seed oil as omega-3 fatty acid (PSO group), 3) basal diet with 2% sunflower oil as omega-6 fatty acid (SFO group), 4) basal diet with 200 mg/kg vitamin E (Control + vitE group), 5) basal diet and 2% pumpkin seed oil along with 200 mg/kg vitamin E (PSO + vitE group) and 6) basal diet and 2% sunflower oil along with 200 mg/kg vitamin E (SFO + vitE group). Roosters were fed on the experimental diets for 60 d and different characteristics of sperm characteristics including routine semen analysis and several sperm functional tests in every 20 d were examined. Reproductive hormones were also evaluated in 0 d and at the end of the trial. Semen volume and morphology were not affected by any of the diets. The roosters fed with pumpkin seed oil + vitE showed the higher percentage of sperm concentration, total motility, progressive motility, viability and membrane integrity and the lower lipid peroxidation ( P ≤ 0.05). The group 5 (PSO + vitE) had numerically the lowest sperm with fragmented DNA (DNA Fr + ) at 0 of the experiment and sperm with non-fragmented DNA (DNA Fr − ) was lowest in group 6 (SFO + vitE) on da 40 the experiment. Testosterone level was not affected by the experimental diets ( P > 0.05), however other hormones (LH and FSH) were affected. Based on the results, the supplementation of aged roosters’ diet with pumpkin seed oil + vitE improves reproductive performance which can be an appropriate strategy to preserve the reproductive performance of aged roosters.