Clinic for Reproduction and Large Animals, University of Ljubljana, Vet Faculty, Ljubljana, Slovenia.
Division of Reproduction, Department of Clinical Sciences, Swedish University of Agricultural Sciences (SLU), Uppsala, Sweden.
Cryobiology. 2019 Feb;86:77-83. doi: 10.1016/j.cryobiol.2018.12.001. Epub 2018 Dec 4.
The cell membrane of ram spermatozoa is more sensitive to the freezing process than in other species due to its composition. As a result, the quality and viability of frozen thawed ram spermatozoa are often poor, which together with the specific structure of the ewe's cervix are the main reasons for lower fertility in ewes after intracervical insemination. In the present study we investigated the effects of semen centrifugation through a single layer of a species-specific colloid (Androcoll-O) on post-thaw quality of ram spermatozoa. Motility, viability and morphology were analysed 0, 6, 12 and 24 h after thawing. DNA fragmentation index (%DFI) of the samples was assessed 0 h after thawing, by SCSA™. Membrane and acrosome integrity of spermatozoa were analysed by Sybr-14/PI/PNA test 0 h after thawing. The proportion of motile spermatozoa was significantly higher in SLC - selected samples in comparison to control (not SLC - selected) samples at 0, 6, 12 (P < 0.001) and 24 h (P < 0.05). The proportion of viable spermatozoa was also significantly higher in SLC - selected samples in comparison to control samples at all times (P < 0.001). The proportion of abnormal acrosomes and morphologically abnormal spermatozoa (MAS) were significantly lower in SLC - selected samples compared to control samples at all times (P < 0.001). Analysis of chromatin stability revealed significantly lower %DFI values in SLC - selected samples compared to control samples (P < 0.001). The SYBR-14/PI/PNA test also revealed significantly better values in SLC - selected compared to control samples (P < 0.05). In conclusion, single layer colloid centrifugation significantly improved post-thaw quality and longevity of ram spermatozoa, making it suitable for artificial insemination initiatives.
绵羊精子的细胞膜由于其组成比其他物种对冷冻过程更敏感。结果，冷冻解冻绵羊精子的质量和活力通常较差，这与母羊颈管的特定结构一起是颈管内授精后母羊生育率较低的主要原因。在本研究中，我们研究了通过单层种特异性胶体（Androcoll-O）对绵羊精子解冻后质量的影响。在解冻后 0、6、12 和 24 小时分析了运动性、活力和形态。在解冻后 0 小时通过 SCSA™评估了样品的 DNA 碎片化指数（%DFI）。在解冻后 0 小时通过 Sybr-14/PI/PNA 试验分析了精子的膜和顶体完整性。与对照（非 SLC- 选择）样品相比，SLC- 选择样品中运动精子的比例在 0、6、12（P < 0.001）和 24 小时（P < 0.05）时显着更高。与对照样品相比，SLC- 选择样品中存活精子的比例在所有时间点都显着更高（P < 0.001）。与对照样品相比，SLC- 选择样品中异常顶体和形态异常精子（MAS）的比例在所有时间点显着更低（P < 0.001）。染色质稳定性分析显示，SLC- 选择样品中的 %DFI 值明显低于对照样品（P < 0.001）。与对照样品相比，SLC- 选择样品中的 SYBR-14/PI/PNA 试验也显示出明显更好的值（P < 0.05）。总之，单层胶体离心显着改善了绵羊精子解冻后的质量和活力，使其适合人工授精。
