P-122: Sperm Intracellular Reactive Oxygen Species Incidence in Ram Fed A Diet with Fish Oil

Background: Basal level of reactive oxygen species (ROS) is essential for sperm acrosomal reaction, hyperactivation, motility, capacitation and fertilization. Ram spermatozoa contain large quantities of polyunsaturated fatty acids in their plasma membranes so are extremely susceptible to ROS induced damage. Materials and Methods: Eight rams (36 ± 5 month of age and with a weight of 64 ± 7 kg) were randomly distributed in control (CTR; n=4) and fish oil (FO; 35 g/d/ ram; n=4) groups. Rams were fed with fish oil regime in associated with constant levels of vitamin E (45 g/d/ram) for 60 days during the physiological breeding season and after a month of feeding; their semen was collected every week. Computer assisted sperm analyzer system (CASA) with Sperm Class Analyzer software was used to evaluate sperm motility and sperm kinetics (system to setup with analysis values of ovine). Intracellular ROS level was also detected using 2′, 7′- dichlorodihydrofluorescein diacetate (DCFH-DA) for hydrogen peroxide (H2O2) and Dihydroethidium (DHE) for superoxide anion (O2-), by flow cytometry. Results: As expected, adding FO in diet significantly improved volume and concentration as well as total sperm output (5.40 vs. 3.38×109 for FO and CTR, respectively) and improves sperm progressive motility (59.2 vs. and 48.8% for FO and CTR, respectively) as compared to the control group. Totally the rate of O2 was lower in FO group compared to the control one and this difference was significant (p>0.05) between 3rd and 4th week of sampling. On the other hand, the rate of H2O2 didn’t indicate significant difference entirely. Conclusion: In this experiment, using Omega-3 source accompanied by vitamin E could improve the quality and progressive motility of ram spermatozoa, also could decrease the incidence of ROS. As poor diets may impair male fertility potential, probably daily dose of vitamin E and Omega- 3 could compensate dietary deficiencies in some way and thereby optimize male fertility potential.