Standerholen Fride Berg, Waterhouse Karin Elisabeth, Larsgard Anne Guro, Garmo Randi Therese, Myromslien Frøydis Deinboll, Sunde Jan, Ropstad Erik, Klinkenberg Geir, Kommisrud Elisabeth
Department of Production Animal Clinical Sciences, Norwegian University of Life Sciences, Oslo, Norway; Department of Natural Sciences and Technology, Hedmark University College, Hamar, Norway.
SpermVital AS, Hamar, Norway.
Theriogenology. 2015 Aug;84(3):413-20. doi: 10.1016/j.theriogenology.2015.03.028. Epub 2015 Apr 1.
To make timing of artificial insemination (AI) relative to ovulation less critical, methods for prolonging shelf life of spermatozoa in vivo after AI have been attempted to be developed. Encapsulation of sperm cells is a documented technology, and recently, a technology in which sperm cells are embedded in alginate gel has been introduced and commercialized. In this study, standard processed semen with the Biladyl extender (control) was compared with semen processed by sperm immobilization technology developed by SpermVital AS in a blind field trial. Moreover, in vitro acrosome and plasma membrane integrity was assessed and compared with AI fertility data for possible correlation. Semen from 16 Norwegian Red young bulls with unknown fertility was collected and processed after splitting the semen in two aliquots. These aliquots were processed with the standard Biladyl extender or the SpermVital extender to a final number of 12 × 10(6) and 25 × 10(6) spermatozoa/dose, respectively. In total, 2000 semen doses were produced from each bull, divided equally by treatment. Artificial insemination doses were set up to design a blinded AI regime; 5 + 5 straws from each extender within ejaculates in ten-straw goblets were distributed to AI technicians and veterinarians all over Norway. Outcomes of the inseminations were measured as 56-day nonreturn rate (NRR). Postthaw sperm quality was assessed by flow cytometry using propidium iodide and Alexa 488-conjugated peanut agglutinin to assess the proportion of plasma membrane and acrosome-intact sperm cells, respectively. In total, data from 14,125 first inseminations performed over a 12-month period, 7081 with Biladyl and 7044 with SpermVital semen, were used in the statistical analyses. There was no significant difference in 56-day NRR for the two semen categories, overall NRR being 72.5% and 72.7% for Biladyl and SpermVital, respectively. The flow cytometric results revealed a significant higher level of acrosome-intact live spermatozoa in Biladyl-processed semen compared to SpermVital semen. The results indicate that the level of acrosome-intact live spermatozoa in the AI dose did not affect the 56-day NRR for the two semen processing methods. In conclusion, this study has showed that immobilized spermatozoa provide equal fertility results as standard processed semen when AI is performed in a blinded field trial, although the immobilization procedure caused increased sperm damage evaluated in vitro compared to standard semen processing procedure.
为了降低人工授精（AI）时间相对于排卵的严格程度，人们尝试开发能够延长人工授精后精子在体内保存期限的方法。精子细胞封装是一项已记录在案的技术，最近，一种将精子细胞包埋于海藻酸盐凝胶中的技术已被推出并实现商业化。在本研究中，在一项盲法田间试验中，将使用Biladyl稀释剂处理的标准精液（对照）与由SpermVital AS公司开发的精子固定技术处理的精液进行了比较。此外，还评估了体外顶体和质膜完整性，并与人工授精生育力数据进行比较以寻找可能的相关性。采集了16头生育力未知的挪威红牛的精液，并将精液分成两份后进行处理。这些样本分别用标准Biladyl稀释剂或SpermVital稀释剂处理，最终每剂量分别含有12×10⁶和25×10⁶个精子。每头公牛共生产2000剂精液，按处理方式平均分配。人工授精剂量的设置旨在设计一种盲法人工授精方案；将十个吸管杯中每份射精样本中来自每种稀释剂的5 + 5根吸管分发给挪威各地的人工授精技术员和兽医。授精结果以56天不返情率（NRR）来衡量。解冻后精子质量通过流式细胞术进行评估，使用碘化丙啶和Alexa 488偶联的花生凝集素分别评估质膜和顶体完整的精子细胞比例。在为期12个月的时间里，共进行了14125次首次人工授精，其中7081次使用Biladyl精液，7044次使用SpermVital精液，这些数据用于统计分析。两种精液类别的56天不返情率没有显著差异，Biladyl和SpermVital的总体不返情率分别为72.5%和72.7%。流式细胞术结果显示，与SpermVital精液相比，Biladyl处理的精液中顶体完整的活精子水平显著更高。结果表明，对于两种精液处理方法，人工授精剂量中顶体完整的活精子水平并未影响56天不返情率。总之，本研究表明，在盲法田间试验中进行人工授精时，固定化精子与标准处理精液的生育力结果相同，尽管与标准精液处理程序相比，固定化程序在体外评估时导致精子损伤增加。
