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研究生:黃以珊
研究生(外文):Yi-Shan Huang
論文名稱:愛德華氏菌親源性分析及IgY抗體製備之研究
論文名稱(外文):Phylogenetic analysis of Edwardsiella tarda preparation of Edw. tarda-IgY
指導教授:蔡信雄蔡信雄引用關係
指導教授(外文):Shinn-Shyong Tsai
學位類別:碩士
校院名稱:國立屏東科技大學
系所名稱:獸醫學系所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2008
畢業學年度:96
語文別:中文
論文頁數:81
中文關鍵詞:愛德華氏症37 kDa外膜蛋白蛋黃免疫球蛋白
外文關鍵詞:Edwardsiellosis37 kDa outer membrane proteinimmunoglobulin yolk
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愛德華氏症是由愛德華氏菌所引起的,養殖鰻魚感染候會造成嚴重之經濟損失。本研究收集鰻魚病例共40場次共20株並以DNAstar、Mega 4軟體進行親源性分析可得知Edw. tarda之基因型具有很高的相似度 ( >99.4% )。由蛋白質電泳的結果可知各個不同株之愛德華氏菌之外膜蛋白在37 kDa之位置都有陽性的反應,並偵測其Rabbit antisera against 37 kDa OMP抗體力價為256倍,此外Rabbit antisera against six Edw. tarda strains抗體力價為64倍,另IgY against Edw. tarda 37kDa OMP抗體力價為4096倍。另將愛德華氏菌蛋黃免疫球蛋白進行口服人工接種試驗,進行為期三週之試驗並使餵飼濃度達5g。再將愛德華氏菌 ( 96-50 ) 之菌量調整為3×109 CFU並混合魚餌料中進行攻毒,由結果可知IgY against Edw. tarda 37 kDa OMP添加於餌料中予鰻魚餵飼,並在口服攻毒後繼續餵飼IgY後,其相對生存百分比分為60%,66.7%。此外,於口服攻毒後餵飼IgY,其相對生存百分比分為50%,75%。將IgY against Edw. tarda 37 kDa OMP與市售之Edw. tarda peptide比較,IgY之相對生存百分比為75%,較市售之Edw. tarda peptide之50% 高,因此可以得知IgY之效果較佳。
Edwardsiella tarda is thought to be the etiological agent of Edwardsiellosis. It cause heavy economic losses in eel. In this study, we sampled 20 isolates from 40 eel farms. PCR was carried out for detection of Edw. tarda using one set of primers 40F/1500R to amplify a 1450 bp fragment of genomic DNA. Using the DNAstar software and Mega 4 to analyze phylogenetic relationship were highly identical ( >99.4% ). Using SDS-PAGE, 37 kDa OMP can be detected from all Edw. tarda strains. The titer of rabbit antisera against 37 kDa OMP was 256X. In addition, the titer of rabbit antisera against other six Edw. tarda strains was 64X, and IgY against Edw. tarda 37kDa OMP was 4096X. The oral inoculation was performed to each eel by introducing the feed mixed with IgY against Edw. tarda 37kDa OMP once per day for three weeks of 5g and inoculated with Edw. tarda ( 96-50 ) at 3×109 CFU. The feed mixed with IgY in challenge from beginning to end, and relative percent survival was 60%, 66.7%. The feed mixed with IgY afetr challenge, and relative percent survival was 50%, 75%. IgY against Edw. tarda 37 kDa OMP’s relative percent survival was 75%, commodification Edw. tarda peptide’s relative percent survival was 50%.
中文摘要...................................................Ⅰ
Abstract..................................................II
謝誌.....................................................IⅤ
目錄.......................................................V
圖表目錄...................................................IX
第1章 前言..................................................1
第2章 文獻回顧..............................................3
第3章 材料方法.............................................20
第4章 結果.................................................31
第5章 討論.................................................61
參考文獻...................................................65
附錄......................................................78
作者簡介...................................................81
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