熱季下水牛冷凍精液製程對解凍後品質之影響

本試驗於熱季（2004年8～9月）進行，探討不同稀釋液、稀釋方法與平衡時間對水牛冷凍精液解凍後，於37℃培養0和3小時，對其精子活動力與經由精子低滲透壓膨脹試驗所測試之精子膜功能完整性之影響。試驗結果顯示，將稀釋液先以不含甘油之稀釋液稀釋，再於30℃添加含甘油之稀釋液時，對精液冷凍－解凍後，於37℃培養0小時後，其精子活動力與精子膜功能完整性之結果，皆極顯著高於一段添加者（分別為57.2±3.4% vs. 51.3±5.5%及48.9±3.4% vs. 43.8±3.2%, P＜0.01）；培養3小時後，亦以兩段添加之精子膜功能完整性結果顯著高於一段添加者（28.0±2.3% vs. 25±3.4%, P＜0.05）。水牛精液於裝填降溫後，於5℃經2小時平衡時間處理，比0小時者，可顯著改善於37℃培養0小時（50.7±4.9% to 57.9±2.9%, P＜0.01）及3小時（24.7±2.6% to 26.4±1.6%, P＜0.05）之精子活動力；亦顯著改善解凍後經培養0小時（44.9±3.4% to 47.9±4.3%, P＜0.05）及3小時（25.5±2.9% to 28.2±2.7%, P＜0.05）之精子膜的功能完整性。比較三羥基甲基氨基甲烷及乳糖為基質的兩種不同稀釋液處理的效果時，精液經解凍後，於37℃培養0及3小時之精子活動力與精子膜功能完整性之評估皆無顯著差異，顯示兩種稀釋液皆可應用於台灣水牛冷凍精液。由本試驗結果得知於製作台灣水牛冷凍精液時，可於30℃下，將精液先以不含甘油之稀釋液稀釋，再添加含甘油稀釋液後，進行裝填與封口，緩慢降溫至5℃後，經2小時的平衡時間，再進行冷凍過程，可得較佳之冷凍－解凍後，於37℃培養0和3小時之精子活動力與低滲透壓膨脹試驗所測試之精子膜功能的完整性，而其冷凍精液解凍後的人工授精授胎率為50%（9/18）。

關鍵字

台灣水牛；冷凍精液；低滲透壓膨脹試驗

並列摘要

The process of preparing frozen semen significantly affects the quality of frozen-thawed semen. The objective of this experiment was to study the effects of various diluents (TYCG and LYG), methods of dilution (one-step and two-step) and equilibration periods (0 and 2 h) on the post-thawed motility and hypo-osmotic swelling test (HOST) response of spermatozoa and the indication of the functional integrity of sperm membrane of buffalo spermatozoa after incubated at 37℃ for 0 and 3 hours. Results showed that both motility and HOST response of post-thawed semen of one-step dilution after being incubated at 37℃ for 0 hr were significantly higher than that of one-step (57.2±3.4 vs. 51.3±5.5% and 48.9±3.4 vs. 43.8±3.2%, respectively; P ＜ 0.01). The HOST response was also significantly higher in the two-step dilution method than one-step after incubating at 37℃ for 3 hours (28.0±2.3 vs. 25.7±3.4%, P ＜ 0.05). After filling, sealing and cooling to 5℃ of straws, semen with equilibration at 5℃ for 2 hours significantly showed a better motility than semen with the 0 hour equilibrating, after incubating the frozen-thawed semen at 37℃ for 0 hour (50.7±4.9 to 57.9±2.9%, P ＜ 0.01) and 3 hours (24.7±2.6 to 26.4±1.6%, P ＜ 0.05). The semen with 2-hour equilibration at 5℃ also significantly showed improvement on the HOST response than 0 hour equilibration, after incubating at 37℃ for 0 hour (44.9±3.4 to 47.9±4.3%, P ＜ 0.05) and 3 hours (25.5±2.9 to 28.2±2.7%, P ＜ 0.05). Comparing the effects of TYCG and LYG diluents, we found there was no significant difference in motility or HOST response between two diluents after incubating the frozen-thawed semen at 37℃ for 0 and 3 hours. Overall, the procedures of diluting, filling, sealing at 30℃, diluting in two steps and equilibrating at 5℃ for 2 hours for buffalo spermatozoa cryopreservation were essential for producing better results in motility and HOST response of post-thaw buffalo spermatozoa after incubating at 37℃ for 0 and 3 hour, and the conception rate was 50% (9/18).

並列關鍵字

Taiwan water buffalo ； Frozen semen ； Hypo-osmotic swelling test

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