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间接ELISA法研究嗜水气单胞菌对鳗鲡表皮黏液的黏附特性
引用本文:林桂芳,鄢庆枇,陈文博,苏永全,覃映雪.间接ELISA法研究嗜水气单胞菌对鳗鲡表皮黏液的黏附特性[J].中国水产科学,2013,20(5):1023-1031.
作者姓名:林桂芳  鄢庆枇  陈文博  苏永全  覃映雪
作者单位:1. 集美大学 水产学院, 福建省高校水产科技与食品安全重点实验室, 福建 厦门 361021;2. 厦门大学 海洋与地球学院, 福建 厦门 361005
基金项目:国家自然科学基金项目(31272699); 福建省自然科学基金项目(2011J06014).
摘    要:

采用间接ELISA法研究菌浓度、孵育时间、温度、pH、阳离子及碳源等因子对嗜水气单胞菌(Aeromonas hydrophila)黏附鳗鲡(Anguilla anguilla)表皮黏液的影响。结果表明, 改良后的间接ELISA法的检测灵敏度约为9.92×104 CFU, 细菌的黏附量随菌浓度的升高而逐渐增大并符合饱和黏附动力学方程: y=0.135ln(x)-0.936(R2= 0.986); 嗜水气单胞菌黏附鳗鲡表皮黏液的最佳条件为: 温度20~28℃, pH 6.2~6.6, NaClMgCl2质量浓度分别为15~25 g/L3 g/L, 孵育时间为150 min。碳源对嗜水气单胞菌的黏附作用有不同程度的影响, 葡萄糖和麦芽糖能显著提高嗜水气单胞菌的黏附量(P<0. 05), 果糖则显著降低嗜水气单胞菌的黏附量(P<0. 05)。以上结果说明, 嗜水气单胞菌对鳗鲡表皮黏液具有较强的黏附作用, 且其黏附作用具有可控性。



关 键 词:嗜水气单胞菌    黏附    间接ELISA    鳗鲡    表皮黏液
修稿时间:2013/9/26 0:00:00

Adhesion characteristics of Aeromonas hydrophila to eel skin mucus by indirect ELISA
LIN Guifang,YAN Qingpi,CHEN Wenbo,SU Yongquan,QIN Yingxue.Adhesion characteristics of Aeromonas hydrophila to eel skin mucus by indirect ELISA[J].Journal of Fishery Sciences of China,2013,20(5):1023-1031.
Authors:LIN Guifang  YAN Qingpi  CHEN Wenbo  SU Yongquan  QIN Yingxue
Institution:1. Fisheries College, Jimei University; Key Laboratory of Science and Technology for Aquaculture and Food Safety, Xiamen 361021, China;2. College of Oceanography and Environmental Science, Xiamen University, Xiamen 361005, China
Abstract:Adhesion to the host mucus, which is closely related to the virulence of pathogenic bacteria, is the first crucial step in most infections. In this paper, the adhesion characteristics of ) skin mucus was investigated by indirect ELISA. can be used as an index to reflect the number of adhering bacteria. Second, we confirmed that the iCFU. The number of a saturation kinetics equation =0.986) was obtained by plotting the number of adhering bacteria against cell density. The factors affecting bacterial adhesion were then investigated. The results revealed that the number of adhering increased with incubation time and reached a peak after 150 min. Temperature was one of the major factors affecting bacterial adhesion, with peak adhesion occurring at 20−28. pH also influenced bacterial adhesion. In a weak acid with a pH of 6.2−6.6, the adhesion ability of The optimum concentrations of Na+ and adhesion were 1.5−2.5 g/L and 3 g/L, respectively. In summary, the optimum conditions for the adhesion of to eel skin mucuswere 20‒28, and 150 min incubation time. In addition, we found that bacterial adhesion was influenced by carbohydrates. Seven kinds of carbohydrates were investigated and the results indicate that glucosehelp to further elucidate bacterial pathogenicity and help
Keywords:Aeromonas hydrophila  adhesion  indirect ELISA  Anguilla anguilla  skin mucus
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