- 學位論文
愛德華氏菌親源性分析及IgY抗體製備之研究
Phylogenetic analysis of Edwardsiella tarda preparation of Edw. tarda-IgY
摘要
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。
並列摘要
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％.