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Effects of a lecithin and catalase containing semen extender and a second dilution with different enhancing buffers on the quality of cold-stored canine spermatozoa

机译:含卵磷脂和过氧化氢酶的精液补充剂和第二次稀释,使用不同的增强缓冲液对犬精子冷藏品质的影响

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摘要

In the present study a diluent containing 0.8% lecithin (Minitube (R), Tiefenbach, G) for the cold storage of canine semen was compared to a Tris-egg yolk extender (TRIS-EY) containing 20% egg yolk. For this purpose, aliquots of 10 mixed ejaculates (main fractions) were either diluted with TRIS-EY or with three lecithin extenders containing 0.8% lecithin with or without catalase and tyrosine. All samples were examined by computer assisted sperm analysis (CASA), chlortetracycline assay (CTC) and flow cytometry, sperm chromatin structure assay (SCSA) and zona pellucida binding assay (ZBA). Samples were then cold stored for 8 d and examinations repeated at days 3 and 8. Measurement in the CASA were repeated daily and prior to measurement, each sample was diluted with each of 4 enhancers with or without acetylcarnitine. The use of an enhancer proved to be essential for all extenders and after 8 d of cooling, progressive motility (P) and viability (V) still averaged > 70% and > 80% with the lecithin extenders containing additives, whereas with TRIS-EY and without additives it was significantly lower (P < 0.05). The percentage of capacitated spermatozoa did not differ between extender groups and there was no significant increase in acrosome reactions (AR) within 3 d. The chromatin status of cells was not changed by cooling within 8 d. The ZBA showed that with additives, significantly more spermatozoa bound to oocytes when a lecithin extender with additives was used (P < 0.05). In conclusion, the lecithin extender containing catalase, conserved P and V during 8 d of cold storage better than the TRIS-EY extender, however, only when an enhancer was used; addition of acetylcamitine to the enhancer did not further improve semen quality. The introduced lecithin extender / enhancer combination is a useful tool for prolonged storage of cooled semen with excellent longevity and binding ability; addition of tyrosine to the extender did not improve semen quality.
机译:在本研究中,将用于冷藏精液的含0.8%卵磷脂的稀释剂(Minitube(R),Tiefenbach,G)与含20%蛋黄的Tris-蛋黄补充剂(TRIS-EY)进行了比较。为此,用TRIS-EY或三种含0.8%卵磷脂的卵磷脂补充剂(含或不含过氧化氢酶和酪氨酸)稀释10种混合射精的等分试样(主要馏分)。通过计算机辅助精子分析(CASA),金霉素测定(CTC)和流式细胞仪,精子染色质结构测定(SCSA)和透明带结合测定(ZBA)检查所有样品。然后将样品冷藏8天,并在第3天和第8天重复检查。每天并在测量前重复CASA中的测量,每个样品分别用4种增强剂(含或不含乙酰肉碱)稀释。事实证明,对于所有补充剂来说,都必须使用增强剂,并且在冷却8天后,含卵磷脂补充剂的渐进运动(P)和生存力(V)仍平均> 70%和> 80%,而TRIS-EY不含添加剂的情况则显着降低(P <0.05)。补充剂组之间有能力的精子百分比没有差异,并且在3天内顶体反应(AR)没有明显增加。在8 d内冷却不会改变细胞的染色质状态。 ZBA表明,使用添加剂时,当使用带有添加剂的卵磷脂增量剂时,与卵母细胞结合的精子明显更多(P <0.05)。总而言之,在冷藏8天后,含有过氧化氢酶的卵磷脂补充剂的P和V保持较好,而TRIS-EY补充剂仅在使用增强剂的情况下保存。向增强剂中添加乙酰氨基胍丁胺并不能进一步改善精液质量。引入的卵磷脂补充剂/增强剂组合物是有用的工具,可长期保存冷却的精液,并具有出色的寿命和结合能力。在增量剂中添加酪氨酸并不能改善精液质量。

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