Cryoprotectant effects of natural honey on spermatozoa quality of pre-freezing and frozen-thawed boar semen
Cryoprotectant effects of natural honey on spermatozoa quality of pre-freezing and frozen-thawed boar semen
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DOI：
10.1093/jas/skac384
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发表时间：
2023-01-03
影响因子：
3.3
通讯作者：
Stewart，Kara R.
中科院分区：
文献类型：
--
作者：
Balogun，Kayode B.;Nicholls，Griffin;Stewart，Kara R.
天然蜂蜜由于其有益的特性已成功地用于保存哺乳动物配子。本研究的目的是确定蜂蜜在稀释液中的含量，以改善猪精液冷冻前的质量，并研究蜂蜜在稀释液和冷冻介质中的含量对猪精液冻融后质量的影响。采用戴手套的方法从6头终末杂交公猪身上采集精液，进行两次试验。实验1是随机区组设计，评估蜂蜜在猪精液稀释剂中的四个包含水平[对照（0 H）-Androhep Plus或Androhep Plus与0.25%、0.50%和0.75%蜂蜜（分别为0.25H、0.50H和0.75H）]。合并射精，根据处理进行等分，并在17 ºC下冷却24 h。该实验的结果用于确定实验中的包含水平。2.试验2为2 ×3析因设计，评价蜂蜜在猪精液稀释液和冷冻液中的包合效果。将来自个体公猪的精液样品在含有或不含蜂蜜的稀释剂中冷却（C 0：Androhep Plus; C1：Androhep Plus + 0.25%蜂蜜）。24小时后，评估精液样品，在乳糖-蛋黄（LEY）培养基和三种冷冻培养基类型中的一种中稀释; F0：93%LEY + 6%甘油+1% Equex-STM糊剂（ESP）; F1：93%LEY+（3%甘油和3%蜂蜜）+1%ESP;和F2：93%LEY + 6%甘油+（0.5%ESP和0.5%蜂蜜）。使用受控速率冷冻机将样品冷冻在0.5 mL吸管中，并储存在液氮中。在exp. 1、0.25H和0.50H均能显著提高猪精液冷冻后的运动力（P= 0.033）和前向运动力（P= 0.001）。然而，选择0.25H用于实验。2.在exp. 2、解冻后运动和前向运动以C 0 F2最高（P< 0.05），但与C1 F2无显著差异。形态正常的细胞和顶体均高于各含蜜量组（P< 0.05）。总之，Androhep Plus中含有0.25%和0.50%的蜂蜜可提高冷却野猪精液样本24小时后的活力和前向活力。添加0.25%蜂蜜的Androhep Plus可维持冷冻保存的公猪精液中较高的正常精子细胞和顶体。在冷冻培养基中用蜂蜜代替50% Equex-STM糊剂可改善解冻后精子活力、前向运动力、正常精子百分比和冷冻保存的猪精液的顶体。
Natural honey has been successfully used in the preservation of mammalian gametes because of its beneficial properties. The objectives of this study were to determine the inclusion level of honey in extender for improving boar semen quality before freezing and to investigate the effects of honey inclusion in extender and freezing media on post-thaw quality of frozen-thawed boar semen samples. Ejaculates from six terminally crossbred boars were collected using the gloved-hand technique for two experiments. Experiment 1 was a randomized block design, evaluating four inclusion levels of honey in boar semen extender [Control (0H)—Androhep Plus or Androhep Plus with 0.25%, 0.50%, and 0.75% honey (0.25H, 0.50H, and 0.75H respectively)]. Ejaculates were pooled, aliquoted according to treatments, and cooled for 24 h at 17 ºC. The results of this experiment were used to determine inclusion levels in exp. 2. Experiment 2 was a 2 x ×3 factorial design, evaluating the inclusion of honey in boar semen extender and freezing media. Semen samples from individual boars were cooled in extender with or without honey (C0: Androhep Plus; C1: Androhep Plus + 0.25% honey). After 24 h, semen samples were evaluated, diluted in lactose-egg yolk (LEY) media, and one of three freezing media types; F0: 93% LEY + 6% glycerol + 1% Equex-STM Paste (ESP); F1: 93% LEY + (3% glycerol and 3% honey) + 1% ESP; and F2: 93% LEY + 6% glycerol + (0.5% ESP and 0.5% honey). Samples were frozen in 0.5 mL straws using a controlled-rate freezer and stored in liquid nitrogen. In exp. 1, 0.25H and 0.50H improved motility (P= 0.033) and progressive motility (P= 0.001) of cooled boar semen. Nevertheless, 0.25H was selected for exp. 2. In exp. 2, post-thaw motility and progressive motility were highest (P< 0.05) in C0F2 but not different from C1F2. Morphologically normal cells and acrosomes were higher with all inclusion levels of honey (P< 0.05). In conclusion, 0.25% and 0.50% inclusion of honey in Androhep Plus improves motility and progressive motility of cooled boar semen samples after 24 h. Supplementing Androhep Plus with 0.25% honey maintains higher normal sperm cells and acrosomes of cryopreserved boar semen. Replacing 50% Equex-STM paste with honey in freezing media improves post-thaw sperm motility, progressive motility, percentage of normal sperm, and acrosome of cryopreserved boar semen.