Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, Cuenca EC010205, Ecuador; Centro Latinoamericano de Formación de Especies Mayores y Menores, CLAFEM, Cuenca, Ecuador.
Laboratorio de Biotecnología de la Reproducción Animal, Facultad de Ciencias Agropecuarias, Universidad de Cuenca, Cuenca EC010205, Ecuador.
Anim Reprod Sci. 2024 Jul;266:107495. doi: 10.1016/j.anireprosci.2024.107495. Epub 2024 May 16.
The study investigated the impact of resveratrol (RES) on bull sperm cryopreservation employing conventional slow (CS) and ultra-rapid (UR) freezing methods on sperm quality and in vitro fertility. Twenty-four ejaculates from four bulls were divided into four groups based on the cryopreservation method and RES addition: CS-RES (n = 80), CS-Co (n = 80), UR-RES (n = 24), and UR-Co (n = 24). The CS freezing involved exposing sperm straws with 5% glycerol to liquid nitrogen (LN) vapors, while UR freezing submerged sperm drops with 100 mM sucrose directly into LN. Overall, sperm kinematic parameters and integrity of plasma and acrosome membranes significantly decreased (P < 0.001) after cryopreservation. Post-thaw values of motilities (total [TM] and progressive [PSM]), velocities (curvilinear and straight-line), beat cross frequency (BCF), and sperm with intact plasma membrane/intact acrosome (PI-/PNA-) were higher (P < 0.05) with CS-RES and CS-Co treatments compared to UR-RES and UR-Co treatments. CS-RES treatment resulted in greater percentages (P < 0.05) of TM, PSM, PI-/PNA-, and fertility (blastocyst rate) than their control, CS-Co; while UR-RES showed higher BCF values (P < 0.05) than its control, UR-Co. Additionally, UR-RES treatment exhibited lower oxidative stress percentages than UR-Co (P < 0.05). This study presents the following conclusions: (1) the CS freezing resulted in better cryosurvival of bull sperm than UR freezing; (2) the RES supplementation to CS freezing medium improved sperm motility, membrane integrity, and fertility; and (3) despite low cryosurvival sperm and fertility, the RES addition to ultra-rapid freezing medium reduced oxidative stress.
本研究采用传统慢速（CS）和超快速（UR）冷冻方法，研究白藜芦醇（RES）对牛精子冷冻保存的影响，评估精子质量和体外受精能力。从 4 头公牛的 24 份精液中，根据冷冻方法和 RES 添加情况将其分为 4 组：CS-RES（n = 80）、CS-Co（n = 80）、UR-RES（n = 24）和 UR-Co（n = 24）。CS 冷冻涉及将含有 5%甘油的精子吸管暴露在液氮（LN）蒸气中，而 UR 冷冻则将含有 100mM 蔗糖的精子滴直接浸入 LN 中。总体而言，冷冻保存后精子运动参数和质膜与顶体膜的完整性显著降低（P < 0.001）。解冻后，运动能力（总运动速度 [TM]和前向运动速度 [PSM]）、速度（曲线和直线）、鞭打频率（BCF）和具有完整质膜/完整顶体（PI-/PNA-）的精子的活力更高（P < 0.05），CS-RES 和 CS-Co 处理组与 UR-RES 和 UR-Co 处理组相比。与对照 CS-Co 相比，CS-RES 处理导致 TM、PSM、PI-/PNA-和生育力（囊胚率）的百分比更高（P < 0.05）；而 UR-RES 则显示出更高的 BCF 值（P < 0.05），高于其对照 UR-Co。此外，UR-RES 处理组的氧化应激百分比低于 UR-Co（P < 0.05）。本研究得出以下结论：（1）CS 冷冻法比 UR 冷冻法更有利于牛精子的冷冻存活；（2）RES 补充到 CS 冷冻液中可提高精子活力、膜完整性和生育力；（3）尽管精子冷冻存活率和生育力低，但 RES 添加到超快速冷冻液中可降低氧化应激。
