The maintenance of high vitality and motility of ram's spermatozoa during storage at low temperatures has a crucial role for successful fertilization. This study evaluates the effect of the natural antioxidant oregonin on ram semen quality, stored at 5 °C for 48 h. Еighteen ejaculates (three repetitions for 6 ejaculates) from three local breed rams, collecting by artificial vagina, with volume > 1 ml, concentration > 1 × 109/ml and mass motility >3.5 were used for chilling. Each ejaculate was separated in two equal parts, diluted with Tris-glucose-glycerol-egg yolk extender with no oregonin or supplemented with 100μМ oregonin until adjustment of the sperm concentration to 200 × 106 cells/ml and stored at 5 °C for 48 h. The semen quality assessment was based on the main kinematic (by CASA analysis), morphological parameters (by BrightVit kit staining) and mitochondrial status (by MitoView staining) of the spermatozoa on 0, 24 and 48 h of storage, and on in vivo fertility test. Oregonin did not impair the morphology and kept sustained motility of ram spermatozoa stored at 5 °C for 48 h. The curvilinear velocity indicated faster movement of the oregonin treated sperms that corresponded with high percent of spermatozoa with active mitochondria in these samples. The fertilizing capacity of spermatozoa was preserved and pregnancy rate in the experimental group was 80% versus 60% in control. In conclusion, this study provides a new data about positive effect of the natural antioxidant oregonin, supplemented to the extender, on chilled ram semen quality, including fertilizing ability.

中文翻译：

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Alnus incana树皮中天然抗氧化剂Oregonin对5°C储存48 h的公羊精液质量的积极影响。

在低温下储存期间，维持公羊精子的高活力和运动性对于成功施肥至关重要。这项研究评估了天然抗氧化剂牛至蛋白对公羊精液质量的影响，该精液在5°C下储存48小时。使用来自三个地方种公羊的淡小精液（3个重复的6个精液），通过人工阴道收集，体积> 1 ml，浓度> 1×109 / ml，质量运动性> 3.5。将每个射精分成两等份，用不含牛至素的Tris-葡萄糖-甘油-蛋黄补充剂稀释或补充100μM牛至素，直至将精子浓度调节至200×106细胞/ ml，并在5°C下保存48小时。精液质量评估基于主要运动学（通过CASA分析），在储存0、24和48小时以及进行体内生育力测试时，精子的形态学参数（通过BrightVit试剂盒染色）和线粒体状态（通过MitoView染色）。Oregonin不会损害形态，并保持在5°C储存48 h的ram精子的持续运动。曲线速度表明在这些样品中，经oregonin处理的精子运动更快，这对应于具有活跃线粒体的高百分比精子。保留了精子的受精能力，实验组的怀孕率是80％，对照组是60％。总而言之，这项研究提供了有关补充补充剂的天然抗氧化剂Oregonin对冷冻公羊精液质量（包括受精能力）的积极作用的新数据。