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固定化与传统冷冻保存的挪威红牛精液的精子三磷酸腺苷含量、活力及生育力比较

Comparison of sperm adenosine triphosphate content, motility and fertility of immobilized and conventionally cryopreserved Norwegian Red bull semen.

作者信息

Berg Halldor Felde, Kommisrud Elisabeth, Bai Godlove, Gaustad Eline Rustad, Klinkenberg Geir, Standerholen Fride Berg, Thorkildsen Lill Therese, Waterhouse Karin Elisabeth, Ropstad Erik, Heringstad Bjørg, Alm-Kristiansen Anne Hege

机构信息

Norwegian University of Life Sciences, Faculty of Veterinary Medicine, Department of Production Animal Clinical Sciences, Oslo, Norway; SpermVital AS, Hamar, Norway.

Inland Norway University of Applied Sciences, Department of Natural Sciences and Technology, Hamar, Norway; SpermVital AS, Hamar, Norway.

出版信息

Theriogenology. 2018 Nov;121:181-187. doi: 10.1016/j.theriogenology.2018.08.016. Epub 2018 Aug 16.

DOI:10.1016/j.theriogenology.2018.08.016
PMID:30165307
Abstract

Estrus detection and timing of AI remains a challenge in cattle breeding. Prolonging spermatozoa lifespan after AI, making sperm cells available over an extended period, could make timing of AI relative to ovulation less crucial and improve fertility. Immobilization of sperm cells by the patented SpermVital technology in an alginate gel will provide a gradual release of spermatozoa after AI. The first aim of this study was to examine fertility, measured as non-return rate after 56 days (NR56), of SpermVital (SV) processed semen with reduced sperm cell number per dose compared to earlier studies, and compare with conventionally processed semen in Biladyl, a proprietary version of the egg yolk Tris semen extender. The second aim was to examine in vitro sperm quality post-thaw and after thermal stress. The third aim was to examine potential correlations between in vitro sperm parameters and NR56. Ejaculates from 16 Norwegian Red young bulls were split in three, processed and cryopreserved as Biladyl semen (B15; 15 million spermatozoa/dose) or by SpermVital technology (SV25; 25 million spermatozoa/dose or SV15; 15 million spermatozoa/dose). 1400 semen doses were produced per bull and distributed throughout Norway for a blinded field trial. Fertility was recorded as NR56 after first AI (N = 7155). Two ejaculates from each bull were randomly selected for in vitro experiments. B15 and SV15 semen samples were analyzed for motility by computer-assisted sperm analysis, viability and acrosome integrity by flow cytometry and ATP content by bioluminescence assay, post-thaw and after thermal stress. The AI trial detected no differences in NR56; least square means being 75.5% (B15), 75.6% (SV25) and 74.8% (SV15) (p > 0.05). There were no differences in total motility and progressive motility post-thaw, however, after three hours incubation at 38 °C, SV sperm motility and progressivity were higher for SV15 than for B15 spermatozoa (p < 0.05). The percentage of acrosome intact live sperm cells was higher for SV15 than B15 spermatozoa at all timepoints analyzed (0 h, 3 h, 24 h, p < 0.05). B15 semen showed a higher ATP level than SV15 at 0 h (p < 0.05), while SV15 sperm cells had higher ATP levels after 3 and 24 h (p < 0.05). No association was detected between in vitro sperm parameters and NR56. In conclusion, SV15, SV25 and B15 semen yielded equal fertility after AI. However, there were differences in sperm quality, as SV15 spermatozoa displayed higher motility, viability and ATP levels after thermal stress than B15 spermatozoa (p < 0.05).

摘要

在牛的繁殖过程中,发情检测和人工授精(AI)的时机选择仍然是一项挑战。延长人工授精后精子的寿命,使精子细胞在更长时间内可用,可能会降低人工授精相对于排卵时间的重要性,并提高繁殖力。通过专利的SpermVital技术将精子细胞固定在藻酸盐凝胶中,将在人工授精后实现精子的逐渐释放。本研究的首要目的是,与早期研究相比,检测每剂量精子数量减少的经SpermVital(SV)处理精液的繁殖力(以56天后的返情率(NR56)衡量),并与Biladyl(蛋黄Tris精液稀释剂的一种专有版本)中常规处理的精液进行比较。第二个目的是检测解冻后和热应激后的体外精子质量。第三个目的是检测体外精子参数与NR56之间的潜在相关性。将16头挪威红牛青年公牛的射精样本分成三份,分别处理并冷冻保存为Biladyl精液(B15;每剂量1500万个精子)或采用SpermVital技术(SV25;每剂量2500万个精子或SV15;每剂量1500万个精子)。每头公牛生产1400剂精液,并在挪威各地进行盲法田间试验。首次人工授精后记录繁殖力为NR56(N = 7155)。从每头公牛中随机选择两份射精样本用于体外实验。对B15和SV15精液样本解冻后和热应激后的活力进行计算机辅助精子分析,通过流式细胞术检测存活率和顶体完整性,通过生物发光测定法检测ATP含量。人工授精试验未检测到NR56的差异;最小二乘均值分别为75.5%(B15)、75.6%(SV25)和74.8%(SV15)(p > 0.05)。解冻后总活力和渐进性活力没有差异,然而,在38°C孵育三小时后,SV15的SV精子活力和前进性高于B15精子(p < 0.05)。在所有分析时间点(0小时、3小时、24小时,p < 0.05),SV15的顶体完整活精子细胞百分比高于B15精子。在0小时时,B15精液的ATP水平高于SV-15(p < 0.05),而SV15精子细胞在3小时和24小时后的ATP水平更高(p < 0.05)。未检测到体外精子参数与NR56之间的关联。总之,SV15、SV25和B15精液在人工授精后产生了相同的繁殖力。然而,精子质量存在差异,因为热应激后SV15精子的活力、存活率和ATP水平高于B15精子(p < 0.05)。

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