臺灣博碩士論文加值系統

精液冷凍技術具有種原保存、延長精子體外存活時間及應用更具彈性等功能，但是在精液冷凍過程中受到稀釋液、冷凍前處理、冷凍保護劑、冷凍-解凍之速率及精子濃度等因素所影響，使精液保存效果有所變化。
本研究之旨在藉由探討稀釋液對精液冷藏及冷凍保存，以及不同冷凍保護劑對精液冷凍保存之影響。
試驗1利用土雞精液分別以Chaudhuri、Lake及Sexton稀釋液稀釋後進行人工授精，結果顯示Lake、Sexton與原精液之受精率分別為91.3％、87.5％及86.3％，三組間無顯著差異，但與Chaudhuri 稀釋組受精率78.2％呈顯著差異(p<0.05)。由此結果得知Lake與Sexton稀釋液較Chaudhuri稀釋液適合作為土雞精液之稀釋液。實驗2採用Lake與Sexton稀釋液稀釋精液，額外添加DMSO(dimethylsulphoxide)冷凍保護劑進行精液冷凍保存。Lake與Sexton兩組之冷凍-解凍精液進行人工授精後，受精率分別為44.1％與23.4％，兩組間呈顯著差異(p<0.05)，顯示以DMSO為冷凍保護劑情況下，土雞精液進行冷凍保存時，Lake稀釋液較適合作為冷凍稀釋液。實驗3分別利用Lake冷凍方式與Tseltin冷凍方式進行土雞精液冷凍保存，結果顯示Lake冷凍方式解凍後精子之活動力、存活能力與人工授精之結果較Tseltin冷凍方式佳，但是由於本試驗Tselutin冷凍方式經過修正，因此Tselutin冷凍方式是否較差仍需更進一步探討。實驗4以Lake稀釋液稀釋土雞精液，分別添加DMSO、DMA(N,N-dimethylacetamide)及DMSO+DMA混合之冷凍保護劑進行精液冷凍保存，解凍後進行人工授精。混合方式之受精率為44.3％，較單獨添加DMSO(24.1％)或DMA( 0％)佳，顯示冷凍保護劑如採用混合方式具有較佳之精液冷凍保存效果。
綜合試驗結果，顯示土雞精液冷藏保存時，利用Lake及Sexton稀釋液具有良好之效果，精液冷凍保存則以Lake稀釋液並添加DMSO+DMA混合之冷凍保護劑具有較佳之精液冷凍保存效果。

Advantages of frozen semen with artificial insemination are preservation of select lines, extend sperm life and enable the widespread of outstanding sires. However, successfully freezing of chicken semen depends on composition of diluents, type and concentration of cryoprotective agents, rate of cooling and thawing semen.
The objective of this study is to investigate influence of extender on storage of native chicken semen in cooling or freezing state, and to modify concentration of cryoprotective agents.
In exp I, semen was diluted 1:3 with Lake, Sexton and Chaudhuri, Undiluted semen was control group. Fertility of exp I was shown 91.3 ％, 87.5 ％, 78.2 ％ and 86.3 ％ respectively, Chaudhuri group was significant lower than other group. Results indicated that Lake and Sexton extender were suitable extenders for Native chicken semen. In exp II, chicken semen was freezed with Lake and Sexton diluents containing DMSO. Fertility of Lake and Sexton group were 44.1% and 23.4%, Sexton group was significant lower than Lake extender containing DMSO for freezing native chicken semen. Results of exp II show that Lake extender containing DMSO was suitable for freezing native chicken semen. In exp III, chicken semen was freezed with Lake and Tseltin method. Results of motility, viability and fertility indicated Tseltin was lower than Lake group, but Tseltin was modified so further research is needed. In exp IV, chicken semen was diluted 1:3 with Lake extender containing DMSO, DMA and a mix of DMSO and DMA respectively, to freeze and store. After thawing, fertility of exp IV was shown 24.1%, 0% and 44.3％ respectively, DMSO and DMA was lower than the mixed group. Results indicated that a mix of cryoprotective agent was best for frozen semen.
Results of all experiments show that the best of native chicken semen extender on cooling state were Lake and Sexton extender. And the best of freezing semen was diluted with Lake extender containing mixed cryoprotective agents.

中文摘要……………………………………………………………….…I
英文摘要………………………………………………………………..III
誌謝……………………………………………………………………...V
目錄…………………………………………………………………......VI
圖次目錄…………………………………………………………..……IX
表次目錄……………………………………………………………..….X
壹、前言………………………………………………………………..1
貳、文獻探討…………………………………………………………..2
一、家禽之生殖系統………………………………………………..2
(一)、雄禽生殖系統……………………………………………2
(二)、雌禽生殖系統…………………………………………....3
(三)、精子於雌禽生殖道內之儲存……………………………5
二、精液……………………………………………………………...8
(一)、精子之產生……………………………………………….8
(二)、精子之物理結構………………………………………...12
(三)、精漿成分及作用………………………………………...14
(四)精液採集之技術…………………………………………...15
三、精液保存之方法………………………………………………..17
(一)、冷藏保存…………………………………………………17
(二)、冷凍保存………………………………………………….23
四、冷凍保護劑………………………………………………………28
(一)、甘油對精液人工授精之影響…………………………….29
(二)、其他滲透性冷凍保護劑………………………………….32
五、擬解決之問題……………………………………………………34
參、材料與方法…………………………………………………………35
一、精液之收集………………………………………………………35
二、稀釋液之配製……………………………………………………37
三、精液之稀釋………………………………………………………37
四、冷凍稀釋液之配製………………………………………………37
五、精液之冷凍-解凍流程…………………………………………..37
六、活動力鏡檢………………………………………………………39
七、存活能力鏡檢……………………………………………………39
八、人工授精…………………………………………………………40
九、種蛋之收集與保存………………………………………………42
十、種蛋之孵化及照蛋………………………………………………42
十一、實驗處理………………………………………………………45
試驗1、新鮮精液以不同稀釋液處理後對精子活動力、
存活能力及人工授精之影響…………………………………..45
試驗2、不同稀釋液對精液冷凍保存之影響…………………45
試驗3、不同前處理方式對精液冷凍保存之影響……………46
試驗4、添加不同冷凍保護劑對精液冷凍保存之影響………47
十二、統計分析………………………………………………………48
肆、結果與討論…………………………………………………………49
伍、結論…………………………………………………………………73
陸、參考文獻……………………………………………………………74
柒、作者簡介……………………………………………………………88

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