- 期刊
台灣土雞精子冷凍保存後之存活率、受精率及其受精卵之孵化率
Survival rate and fertility of cryopreserved fowl spermatozoa and hatchability of the fertilized eggs
摘要
生殖細胞的保存是一項有效保存遺傳資源的方法。本研究的目的是針對台灣土雞遺傳資源的保存，而研發精子的急速冷凍技術。利用畜產試驗所保有的台灣土雞，採集5隻公雞的精液後，將其混合，再加入等量的12% dimethylacetamide（DMA）抗凍液，就置於冰塊間於0℃平衡10分鐘後，立即以微量滴管將含有抗凍劑的精液以小滴（約20～25μl）的形態滴入液態氮中製成粒狀精液，並將12～15粒的冷凍精液裝入冷凍小瓶保存。抗凍液是以市售豬精液稀釋用速保精溶液來溶入12% DMA。粒狀冷凍精液的解凍步驟是先將0.4 ml的速保精溶液裝入玻璃小瓶，置於60℃恆溫水槽中1～2分鐘後取出，再將冷凍小瓶內的粒狀精液迅速倒入玻璃小瓶混合，隨即將解凍後的精液置於冰塊間，攜至現場進行母雞人工授精測試。分別在熱季（七月）與涼季（一月）進行測試。授精時間為下午3點，共60隻母雞分為A、B兩組各30隻，A組為每天授精一次，連續授精3天；B組為僅授精一次。解凍精子的存活檢測是以SYBR－14和PI螢光染劑進行，解凍後精子的存活率為45－50%。授精後1－7天撿蛋，孵蛋後的第7天進行照蛋檢查受精率；熱季之受精率與受精蛋孵化率分別為A組50%（40/80）與95%（38/40）、B組20%（14/69）與86%（12/14）；涼季之受精率與受精蛋孵化率分別為A組90%（73/81）與96%（70/73）；B組70%（53/76）與92%（49/53）。依據本試驗結果顯示以粒狀冷凍精液進行台灣土雞的種原保存是可行的，但建議以涼季來進行種原繁衍會有較好的授精率。
並列摘要
Cryopreservation of germ cells is an efficient method to preserve genetic resources. This study was conducted to develop a rapid freezing method to preserve the spermatozoa of Taiwan native chicken. Semen of five roosters from TLRI were collected and mixed, and then diluted with freezing medium containing 12% dimethylacetamide(DMA)on an equal volume basis. Diluted semen were then kept in 0℃ ice bath for 10 minutes and then the diluted semen were made into droplet (20~25 μl per drop) and put directly in liquid nitrogen. The semen pellets were packed in 1 freezing vial per 12-15 pellets for the storage in liquid nitrogen. Freezing medium was made from a base medium of commercial pig semen extender (Sperm-Up solution) with 12% DMA. Semen was collected, frozen in hot and cool season, respectively. Thawing the semen pellets was performed with 0.4 ml Sperm-Up solution pre-heat to 60℃ per vial, and then kept the vial on 0℃ ice bath till insemination. Insemination was performed 30 hens of group A for three consecutive days or in 30 hens of group B for a single insemination. Egg was collected from 1^(st) until 7^(th) day after insemination. The survival rate of fowl spermatozoa varied from 45 to 50% in this study. Fertilities were 50% (group A) and 20%(group B)in hot season, and those of cool season were 90% (group A) and 70%(group B) , respectively. Hatchabilities were 95% (group A) and 86%(group B) respectively in hot season, and those of cool season were 96% (group A) and 72% (group B), respectively.