Rapid Identification of Pathogenic Fich Bacteria

致病性菲鱼细菌的快速鉴定

• 批准号: 04556027
• 负责人: KOBAYASHI Masanori
• 金额: $ 4.99万
• 依托单位: Kitasato University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research (B)
• 财政年份: 1992
• 资助国家: 日本
• 起止时间: 1992 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-04556027/
• 关键词: Identification; Pathogenic Fish Bacteria; Renibacterium salmoninarum; Yellowtail; Coho Salmon; Western Blotting Assay; Dot Blotting Assay; Fluorescent Antibody Technique; Renibacterinm salmoninarum; Pasteurella Piscicida; 魚病細菌; 魚類; 同定法

The API ZYM system was used to identify the bacterial fish pathogens. As additional test, Gram stain and the activities of cytochrome oxidase and catalase were examined. The results of API ZYM and the additional tests were coded for easy identification.Various diagnostic methods used to detect Renibacterium salmoninarum, the causative agent of bacterial kidney disease (BKD) , were compared. The most sensitive method was the enzyme immunoassay, which could detect 10^2 bacterial cells per gram of kidney tissue. The minimum bacteria concentrations showing posotive reactions to the indirect fluorescent antibody test (IFAT) and the coagglutination test were 10^3 and 10^4 cells/g kidney, respectively. The sensitivities of the Gram stain, immunodiffusion procedure, and latex agglutination test were low, and these methods could only be applied to fish with overt BKD symptoms. Altogether, 656 coho salmon Oncorhynchus kisutch were examined for R.salmoninarum antigen with the direct and indirect dot blot assays (DDBA and IDBA) and the IFAT.Among the fish sampled, positive reactions were obtained in 11.8% by the DDBA, 28.2% by the IDBA, and 12.9% by the IFAT.

采用API Zym系统对细菌性鱼类病原菌进行鉴定。作为补充试验，检测革兰氏染色和细胞色素氧化酶、过氧化氢酶的活性。为了便于识别，对API Zym和附加试验的结果进行了编码，并对检测细菌性肾脏病(BKD)的病原体沙门氏菌的各种诊断方法进行了比较。最灵敏的方法是酶免疫法，每克肾组织可以检测出10^2个细菌细胞。间接荧光抗体试验(IFAT)和凝集试验阳性反应的最低细菌浓度分别为10^3和10^4个/g肾。革兰氏染色、免疫扩散法和乳胶凝集试验的敏感性较低，这些方法仅适用于有明显BKD症状的鱼类。用直接和间接斑点杂交法(DDBA和IDBA)和IFAT对656尾鲑鱼进行了沙门氏菌抗原检测，其中DDBA法阳性率为11.8%，IDBA法阳性率为28.2%，IFAT法阳性率为12.9%。

项目成果

酒井 正博,厚田 静男 小林 正典: "養殖ギンザケの各飼育期におけるRenibacterium Salmoninarum抗原の検出" 水産増殖. 40. 111-114 (1992)

Masahiro Sakai、Shizuo Atsuta、Masanori Kobayashi：“养殖银大麻哈鱼每个饲养期的鲑鱼肾杆菌抗原的检测”渔业增殖。 40. 111-114 (1992)

Sakai,M.Atsuta,S.Kobayashi,M.: "Susceptibility of five salmonid fishes to Renibacterium salmoninarum" Gyobyo Kenkyu. 26. 159-160 (1991)

Sakai，M.Atsuta，S.Kobayashi，M.：“五种鲑鱼对鲑鱼肾杆菌的敏感性”Gyobyo Kenkyu。

Sakai,M.Kobayashi,M.: "Detection of Renibacterium salmoninarum,the causative agent of bacterial kidney diseasein salmonid fish,from pen-cultured coho salmon" Appl Environ Microbiol. 58. 1061-1063 (1992)

Sakai，M.Kobayashi，M.：“从圈养银鲑鱼中检测鲑鱼肾杆菌，鲑鱼细菌性肾病的病原体”Appl Environ Microbiol。

Sakai,M.Soliman,M.K.Yoshida,T.Kobayashi,M.: "Identification of Pathogeric Fish Bacteria Using the Apl ZYM System" Can.J.Fish.Aquat.Sci.50. 1137-1141 (1993)

Sakai，M.Soliman，M.K.Yoshida，T.Kobayashi，M.：“使用 Apl ZYM 系统鉴定鱼类病原细菌”Can.J.Fish.Aquat.Sci.50。

Kawahara,E.Inarimori,T.Nomura,S.Takahashi,Y.: "Humoral immune response of white-spotted char Salvelinus Lencomaenis to Aeromonas salmonicida extracellular products" Nippon Suisan Gakkaishi. 57. 1057-1063 (1991)

Kawahara，E.Inarimori，T.Nomura，S.Takahashi，Y.：“白斑红点鲑对杀鲑气单胞菌细胞外产物的体液免疫反应”Nippon Suisan Gakkaishi。