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石莼提取物对公牛精子冷冻保存及后续胚胎发育的影响。

The effect of Caulerpa sertularioides extract on bull sperm freezablity and subsequent embryo development.

机构信息

Theriogenology Section, Department of Clinical Science, Faculty of Veterinary Medicine, University of Tabriz, Tabriz, Iran.

Faculty of Agriculture, Department of Veterinary Medicine, Shoushtar Branch, Islamic Azad University, Shoushtar, Iran.

出版信息

Theriogenology. 2022 Sep 1;189:167-176. doi: 10.1016/j.theriogenology.2022.06.017. Epub 2022 Jun 20.

Abstract

Artificial insemination is a valuable and essential tool in genetic improvement programs, and its success requires proper semen collection, freezing, and thawing procedures. Nowadays, despite applying of advanced protocols for semen cryopreservation, post-thawing sperm quantitative and qualitative parameters are not satisfactorily comparable to fresh sperm. The present study was designed to evaluate the effects of the supplementation of an alcoholic extract of Caulerpa sertolarioides alga into the tris-egg yolk-based Simmental bull sperm freezing media. The pooled semen samples were divided into five groups, of which four were supplemented with 500, 1000, 1500, and 2000 ppm alga extract and one allocated as a control. Total motility, progressive motility, plasma membrane integrity, DNA integrity, apoptosis, superoxide dismutase (SOD) and glutathione peroxidase (GPx) activities, and total antioxidant capacity (TAC) of sperm were measured. The frozen sperm from each group were used for IVF on the slaughterhouse-derived oocytes. Fertilization, cleavage, and blastocyst rates were assessed for all groups. Total motility, progressive motility, and velocity curvilinear (VCL) parameters were higher (p ≤ 0.05) in group 1000 ppm than the control group. Velocity in a straight path (VSL) was higher (p ≤ 0.05) in all treatment groups except in 500 ppm compared to the control group. Average path velocity (VAP) was higher (p ≤ 0.05) in 1000 and 1500 ppm groups than in the control group. Straightness (STR) showed a higher value (p ≤ 0.05) in 1000 and 2000 ppm than the control group. Groups 500 and 1000 ppm showed more viable sperm than the control group (p ≤ 0.05). DNA damage was lower (p ≤ 0.05) in group 1000 ppm than in the control group. HOST was higher (p ≤ 0.05) in all groups than in the control group. SOD, GPx, and TAC were higher (p ≤ 0.05) in 1000 ppm than the control and all other groups. Apoptosis was not significantly different among the treatment and control groups. In conclusion, supplementation of alcoholic extract of Caulerpa sertularioides into the Simmental bull freezing extender ameliorated the sperm parameters after the freeze-thawing process. Moreover, the results of this study indicated that the best dose to achieve the antioxidant properties of the alga extract in Simmental bull sperm freezing media was 1000 ppm. It was also evident that 1000 ppm alga extract supplementation into the bull sperm improved fertilization, cleavage, and blastocyst rates.

摘要

人工授精是遗传改良计划中非常有价值和必不可少的工具,其成功需要适当的精液采集、冷冻和解冻程序。如今,尽管应用了先进的精液冷冻保存方案,但解冻后精子的数量和质量参数与新鲜精子相比并不令人满意。本研究旨在评估在基于三卵黄的西门塔尔公牛精子冷冻培养基中添加酒精提取的鹿角藻的效果。将混合精液样本分为五组,其中四组分别添加 500、1000、1500 和 2000ppm 海藻提取物,一组作为对照。测量精子的总活力、前向运动活力、质膜完整性、DNA 完整性、凋亡、超氧化物歧化酶(SOD)和谷胱甘肽过氧化物酶(GPx)活性以及总抗氧化能力(TAC)。每组的冷冻精子均用于屠宰场来源的卵母细胞的体外受精。评估所有组的受精、卵裂和囊胚率。与对照组相比,1000ppm 组的总活力、前向运动活力和曲线速度(VCL)参数更高(p≤0.05)。除 500ppm 组外,所有处理组的直线速度(VSL)均较高(p≤0.05)。平均路径速度(VAP)在 1000 和 1500ppm 组中高于对照组(p≤0.05)。直线度(STR)在 1000 和 2000ppm 组中高于对照组(p≤0.05)。与对照组相比,500 和 1000ppm 组的存活精子更多(p≤0.05)。1000ppm 组的 DNA 损伤低于对照组(p≤0.05)。HOST 在所有组中均高于对照组(p≤0.05)。1000ppm 组的 SOD、GPx 和 TAC 均高于对照组和其他组(p≤0.05)。凋亡在处理组和对照组之间没有显著差异。总之,在西门塔尔公牛冷冻液中添加酒精提取的鹿角藻可改善冷冻-解冻过程后的精子参数。此外,本研究结果表明,在西门塔尔公牛精子冷冻液中达到海藻提取物抗氧化特性的最佳剂量为 1000ppm。显然,在公牛精子中添加 1000ppm 海藻提取物可提高受精率、卵裂率和囊胚率。

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