稀釋液種類對乳用種公牛精液冷凍及冷藏保存後精子品質之影響

Diluent Effect on Dairy Bull Post-thawed and Refrigerated Liquid Spermatoza Quality

趙俊炫(Jiun-Shuan Chao) ； 陳一明(I-Ming Chen) ； 陳宜鴻(I-Hung Chen) ； 陳怡璇(Yi-Hsuan Chen) ； 康定傑(Ting-Chieh Kang) ； 陳裕信(Yu-Hsin Chen) ； 曲鳳翔(Fung-Hsiang Chu) ； 李國華(Kuo-Hua Lee) ； 蕭振文(Jen-Wen Hsiau)

《台灣農學會報》 20卷1期 (2019/03) Pp. 59-69

https://doi.org/10.6730/JAAT.201903_20(1).0005

摘要

本研究旨在建立乳用種公牛精液冷凍或冷藏保存時較佳之稀釋液種類。本試驗係採用2頭成熟乳公牛，以假陰道法採取新鮮精液，測定採集精液之體積、濃度及活力，並分別以Tris－檸檬酸－葡萄糖（Tris-citric acid-glucose；TCG）-15%蛋黃、TCG-5% LDL（低密度脂蛋白）、TCG-10% LDL、Biladyl-15%蛋黃、Biladyl-5% LDL、Biladyl-10% LDL及BIOXcell等7種精液稀釋液稀釋，加以探討精液冷凍於液態氮後解凍之精子活力的影響。結果顯示，冷凍精液解凍後之精子存活，以Biladyl-10% LDL及BIOXcell稀釋液冷凍者較佳，可達70%以上解凍活力；利用Biladyl-10% LDL稀釋液冷凍精液進行經產母牛人工授精配種後至少有5成以上懷孕率。在冷藏精液部分，以Biladyl-10% LDL稀釋液稀釋，評估冷藏天數對精子活力及受孕力之結果發現，精液冷藏保存3日其精子仍具有受孕力。綜上所述，本研究已建立完備荷蘭乳公牛採精及精液保存流程，並建議以稀釋液Biladyl-10% LDL及BIOXcell等稀釋液進行乳公牛精液冷凍保存；若以冷藏保存則至少可保存3日，此等結果將可利用已適應本土氣候之菁英公牛進行臺灣母牛群之育種改良。

關鍵字

乳用公牛 ； 稀釋液種類 ； 冷凍保存 ； 冷藏保存 ； 精子品質

並列摘要

This study investigates the diluent effect on dairy bull liquid spermatozoa motility after thawing and refrigeration. Seven different semen diluents including TCG-15% egg yolk, TCG- 5% LDL, TCG-10% LDL, Biladyl-15% egg yolk, Biladyl-5% LDL, Biladyl-10% LDL and BIOXcell were assessed for post-thaw motility. The results show that with the commercial extenders Biladyl-10% LDL and BIOXcell, the post-thawing spermatozoa motility is improved by more than 70%. Thus the pregnancy rate following artificial insemination could be improved by 50% when using post-thawed semen with Biladyl-10% LDL diluent. Refrigerated liquid semen with Biladyl-10% LDL diluent was assessed for spermatozoa motility and calving rate after artificial insemination. It was found that spermatozoa kept for 3 days at 4°C retain fertilization ability. In conclusion, Biladyl-10% LDL or BIOXcell diluent would be recommended for dairy bull frozen semen production. Application of freezing and cooling techniques to bull semen derived from heat-tolerant individuals could facilitate a cattle breeding program to improve Taiwan's dairy industry.

並列關鍵字

Holstein bull ； diluents ； cryopreservation ； refrigerated storage ； spermatozoa quality

參考文獻

Babiak, I., J. Glogowski, M. J. Luczynski, M. Luczynski, and W. Demianowicz. 1999. The effect of egg yolk, low density lipoproteins, methylxanthines and fertilization diluent on cryopreservation efficiency of northern pike (Esox lucius) spermatozoa. Theriogenology 52(3):473-479. doi: 10.1016/S0093-691X(99)00144-2

Barbas, J. P., and R. D. Mascarenhas. 2009. Cryopreservation of domestic animal sperm cells. Cell Tissue Bank. 10(1):49-62. doi: 10.1007/s10561-008-9081-4

Barszcz, K., D. Wiesetek, M. Wasowicz, and M. Kupczynska. 2012. Bull semen collection and analysis for artificial insemination. J. Agri. Sci. 4(3):1-10. doi: 10.5539/jas.v4n3p1

Bergeron, A., M. H. Crete, Y. Brindle, and P. Manjunath. 2004. Low-density lipoprotein fraction from hen's egg yolk decreases the binding of the major proteins of bovine seminal plasma to sperm and prevents lipid efflux from the sperm membrane. Biol. Reprod. 70(3):708-717. doi: 10.1095/biolreprod.103.022996

Bergeron, A., and P. Manjunath. 2006. New insights towards understanding the mechanisms of sperm protection by egg yolk and milk. Mol. Reprod. Dev. 73(10):1338-1344. doi: 10.1002/mrd.20565