沈添富2006-07-262018-06-292006-07-262018-06-292003http://ntur.lib.ntu.edu.tw//handle/246246/16452本試驗之目的在探討菜鴨因選拔及週齡所造成蛋殼品質下降之機制。鴨隻選留是以行政院農委會畜試所宜蘭分所之L105 品系菜鴨，以其40 週齡蛋殼強度為指標，以人工授精方式進行高蛋殼強度(HES)及低蛋殼強度(LES)品系第一代及第二代之繁殖。試驗一為當第一代鴨隻達27、49 及71 週齡時，測定蛋重、蛋殼強度、蛋殼厚度及分析蛋殼中鈣、鎂及有機基質之含量，並比較不同蛋殼強度及不同週齡間蛋殼中成分之差異;此外，將雞蛋殼蛋白質抗體與鴨蛋殼蛋白質進行交互反應偵測，以了解鴨蛋殼蛋白質之特性。試驗二犧牲第二代28、49 及71 週齡之HES 及LES 鴨隻，比較此兩族群鴨隻之血液及子宮液中成分之差異，並將雞蛋殼蛋白質抗體與鴨子宮液蛋白質進行交互反應偵測，以了解鴨子宮液蛋白質之特性。試驗三為刮取HES 及LES 菜鴨殼腺黏膜細胞，並萃取總RNA，以差異顯現反轉錄-聚合　鏈鎖反應，進行其基因表現差異之研究。 經蛋殼強度雙向選拔後，無論第一代或第二代，HES 族群之蛋殼強度顯著較LES 者為高。第一代菜鴨之蛋殼鈣含量在兩族群間並無明顯差異。蛋殼鎂含量則以LES 較高，且此趨勢持續至71週齡。蛋殼中有機基質含量兩族群間無顯著差異，但有機基質含量似隨週齡增加而有增加的趨勢。兩族群蛋殼中蛋白質經SDS-PAGE 電泳後發現，71 週齡之LES有兩蛋白質之表現量似較HES者為高，其分子量分別約80 及180 kDa;然而在27 及49 週齡，兩族群間之蛋殼蛋白質並無差異存在。當探討週齡間之蛋殼蛋白質表現差異發現，180 kDa 蛋白質在71 週齡LES 蛋殼中表現較明顯，但在27 及49 週齡則表現甚微。菜鴨蛋殼蛋白質可與雞蛋殼蛋白質抗體anti-ovocleidin-17 及anti-ovocleidin-116產生交互反應;與anti-ovocleidin-17 有交互反應之蛋白質有兩種，分別為15及18 kDa，其中15 kDa 蛋白質N-端定序之結果得到八個胺基酸殘基NKYPKGWL。 第二代HES 菜鴨血液pH 有較LES者高之趨勢，但血漿總鈣濃度以LES 菜鴨較高，此血液pH 及血漿總鈣濃度之趨勢在三種週齡皆相同。血漿總鎂濃度則HES 及LES 鴨隻間無差異。子宮液pH 及總鎂濃度在HES 及LES 鴨隻間無差異，但HES 菜鴨子宮液總鈣濃度在某些時間點顯著較LES 者為高。HES 鴨隻之子宮液體積在三種週齡皆有較LES 者為高的趨勢，但個體間之變異極大。菜鴨子宮液蛋白質與雞蛋殼蛋白質抗體anti-ovocleidin-17 及anti-ovocleidin-116亦可產生交互反應;子宮液蛋白質之SDS-PAGE 結果顯示：菜鴨在產蛋後不同時間點，其子宮液之蛋白質種類有所不同，但HES 及LES 子宮液蛋白質表現量之差異不顯著。差異顯現反轉錄聚合　鏈鎖反應分析結果顯示，HES 及LES 菜鴨殼腺黏膜細胞之基因表現有差異，部分具蛋殼強度專一性的序列已完成選殖及定序。 本試驗已完成不同蛋殼強度及不同週齡菜鴨之蛋殼、血液、子宮液成分差異之探討，並探討不同蛋殼強度菜鴨之殼腺黏膜細胞基因表現，研究成果將提供改善蛋殼品質之參考。The objectives of this study were to investigate the mechanism of deterioration of eggshell quality associated with age and selection. The 1st generation (F1) and 2nd generation (F2) of Tsaiya ducks laying high eggshell strength (HES) and low eggshell strength (LES) were produced through selection. Trial 1: When F1 ducks reached 27, 49, and 71 weeks of age, egg weight, eggshell strength, and eggshell thickness were collected. Eggs collected were used to analyze the composition of eggshell, including contents of calcium, magnesium, and organic matrix. Proteins in the eggshell were also analyzed and compared by electrophoresis. Trial 2: The blood and uterine fluid of F2 ducks at 28, 49, and 71 weeks of age were collected to compare pH, Ca and Mg concentrations in the blood and uterine fluid between HES and LES ducks. In addition, volume of uterine fluid and proteins in the uterine fluid were also studied. The results showed that eggshell strength of eggs from HES and LES were significantly different for both generations. The eggshell calcium content was similar between HES and LES populations. The significantly higher eggshell Mg content in the LES duck eggshell was observed throughout this study. Organic matrix contents in the eggshell of HES and LES were similar, but they tended to increase with age. The molecular weights of eggshell proteins were between 6.5 and 200 KDa as determined by electrophoresis. Two proteins of them, 80 and 180 KDa, appeared to be associated with eggshell quality. The eggshell of 71-week-old ducks seemed to have more 180 kDa protein than that of 27-week-old ducks. Two proteins in the duck eggshell were able to show cross-reaction with hen’s anti-ovocleidin-17 and anti-ovocleidin-116, respectively. Eight amino acid residues of NKYPKGWL were obtained through N-terminal sequencing of the 15 kDa protein in the duck eggshell. The HES ducks had a trend of higher blood pH than the LES ones. However, the LES ducks had higher levels total blood calcium levels. The tendencies for blood pH and total calcium were similar in the three ages. Total blood magnesium levels were similar between HES and LES ducks. In respect of uterine fluid, pH and total magnesium levels were similar between HES and LES ducks. However, total calcium concentrations were higher in the HES ducks at some time points than in the LES ones. A trend of higher uterine fluid volume was observed in the HES ducks in three ages. Two proteins in the duck’s uterine fluid were able to show cross-reaction with hen’s anti-ovocleidin-17 and anti-ovocleidin-116, which means non-specific binding existing. SDS-PAGE of proteins in uterine fluid showed protein profiles were different among different time points. No significant differences were observed in proteins of uterine fluid between HES and LES ducks based on the SDS-PAGE results. Some eggshell strength-specific bands were observed through differential display reverse transcription-polymerase chain reaction. Two of them had been cloned and sequenced. This study has demonstrated the difference of contents in the eggshell, blood, and uterine fluid of HES and LES ducks. Furthermore, gene expression of shell gland mucosa of HES and LES ducks has also been studied. These data would be helpful to provide some information in solving the problems of eggshell quality deterioration.application/pdf195848 bytesapplication/pdfzh-TW國立臺灣大學動物科學技術學系暨研究所菜鴨蛋殼強度鈣鎂蛋白質基因表現Tsaiya duckEggshell strengthCalciumMagnesiumProteinGene expressionreporthttp://ntur.lib.ntu.edu.tw/bitstream/246246/16452/1/912313B002371.pdf