Abstract

The problems of fertility reducing of rooster semen in the cycle “native sperm–equilibration–short-term and long-term storage (cryopreservation)” are urgent. The purpose of this study was to determine the effect of different methods of preparation (centrifugation or filtration) of rooster semen on its quality characteristics, depending on the method of removing possible pollutions; to evaluate the change in the composition of the cytosol of spermatozoa of native sperm under the influence of dilution and during short-term storage. Materials and methods. Semen of roosters (n = 22) of the Russian white breed was used. Experiment 1: semen was divided into 3 aliquots: I—diluted with synthetic cryoprotective medium LCM in a ratio of 1:1, II—filtered semen diluted with medium (membrane pore diameter 0.2 μm), III—centrifuged (at 3000 rpm in for 10 minutes). Native and frozen/thawed sperm were evaluated in terms of damage to spermatozoa membranes, chromatin, and acrosomes. The composition of carbohydrates and polyols of native spermatozoa was assessed under the influence of dilution and after storage (3 h). The advantage of filtration as a method of technological preparation of semen compared to centrifugation in terms of progressive motility (with rectilinear-translational movement) of sperm (41.0% versus 27.0%) and chromatin damage (43.4% versus 66.4%) has been shown. The same advantage was observed in frozen/thawed sperm filtered before freezing in terms of progressive motility (25.5% vs. 5.5%) and chromatin damage—16.5% vs. 33.6%, respectively. Semen filtration, as a method of technological processing of rooster semen, can be an effective additional step in the preparation of semen for artificial insemination and/or short-term storage. The main component in the composition of the cytosol of native spermatozoa, according to the content of carbohydrates and polyols, was inositol —73.7% of Σ carbohydrates and polyols. The level of inositol decreased during storage by 6.5 times (from 0.030 to 0.007 mg/mL). The data obtained let us suppose the role of inositol as the main antioxidant in the cytosol of spermatozoa. Technological factors of storing rooster semen in various modes (short-term at a temperature of 5°C and long-term at a temperature of –196°C) have a significant impact on the ratio of sperm cytosol components (carbohydrates and polyols). A significant, 2.5‑fold decrease in the relative content of inositol in the cytosol of frozen/thawed spermatozoa, compared with the indicators of native semen, allows us to recommend the introduction of the antioxidant inositol into the composition of cryoprotective media for rooster semen.

中文翻译：

---

工艺因素影响下公鸡精子动力学参数及细胞学特征的变化

摘要

公鸡精液在“天然精子-平衡-短期和长期储存（冷冻保存）”循环中的生育力降低问题迫在眉睫。本研究的目的是确定公鸡精液的不同制备方法（离心或过滤）对其质量特性的影响，具体取决于去除可能污染的方法；评估在稀释的影响下和短期储存期间天然精子的精子细胞质组成的变化。材料和方法。使用俄罗斯白公鸡（ n = 22）的精液。实验1：精液分为3等份：I—用合成冷冻保护介质LCM按1:1的比例稀释，II—过滤后的精液用介质（膜孔径0.2μm）稀释，III—离心（3000rpm离心） 10分钟）。对天然精子和冷冻/解冻精子的精子膜、染色质和顶体损伤进行了评估。在稀释的影响下和储存后（3小时）评估天然精子的碳水化合物和多元醇的组成。与离心相比，过滤作为精液技术制备方法在精子渐进运动（直线平移运动）（41.0% 对 27.0%）和染色质损伤（43.4% 对 66.4%）方面具有优势。在冷冻前过滤的冷冻/解冻精子中也观察到了同样的优势，即前进活力（25.5% 与 5.5%）和染色质损伤（分别为 16.5% 与 33.6%）。精液过滤作为公鸡精液的技术处理方法，可以成为人工授精和/或短期储存精液制备中的有效附加步骤。天然精子细胞质的组成中，根据碳水化合物和多元醇的含量，主要成分是肌醇——占Σ碳水化合物和多元醇的73.7%。储存期间肌醇含量降低了 6.5 倍（从 0.030 降至 0.007 mg/mL）。获得的数据让我们假设肌醇作为精子细胞质中主要抗氧化剂的作用。公鸡精液以各种方式保存的技术因素（5℃短期保存和–196℃长期保存）对精子细胞质成分（碳水化合物和多元醇）的比例有显着影响。与天然精液的指标相比，冷冻/解冻精子的细胞质中肌醇的相对含量显着降低了 2.5 倍，因此我们建议将抗氧化剂肌醇引入公鸡精液冷冻保护介质的成分中。