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To achieving high efficiency in sheep, ram sperm must have high fertilization ability. Quercetin, a potent antioxidant, has been shown to have a positive effect on semen in different species. This study aims to improve the efficiency of the Merino ram sperm using quercetin. The ejaculates collected from Merino rams were used for the dilution of the sperm (sperm density ≥ 3×109 spermatozoa/ml, motility ≥ 75%). Tris diluent was used to dilute the semen. The ejaculates were divided into 4 equal volumes at 32 °C. Three groups were formed by adding quercetin (15, 30, 45 μg/mL) at different doses. Antioxidant (quercetin) did not participate in the 4th group (the control group). Spermatozoa were diluted and frozen to approximately 4x108 spermatozoa per ml with Tris diluent. After thawing, the master mix was prepared with 60μl semen, 10μl FITC-PNA and 2.5μl PI and allowed to incubate at 37 °C for 20 minutes. The reaction was then terminated after treatment with 10μL Hancock solution. An amount of 2.5μl of the sample was taken and 200 sperm were counted at 400 magnifications on a fluorescent microscope. Sperm cells with a green color were evaluated as damaged acrosome, and no green colour was regarded as undamaged acrosome. In terms of the undamaged percentage of acrosomal integrity was assessed in the control group and quercetin groups (15, 30, and 45μg/mL). Findings: 39.16 ± 2.65b in the 15μg/mL group, 40.83 ± 0.30b in the 30μ/mL group, 20.83 ± 2.95a in the 45μg/mL group and 54.83 ± 3.38c in the control group. The percentage of the three groups formed with the quercetin substance added at different doses (15, 30, 45μg/mL) were lower than the control group. Quercetin has been shown to toxicize acrosome integrity at these doses.

Investigation of the Effectiveness of Quercetin on Acrosome Integrity of Merino Ram Sperm