河流弧菌对牙鲆表皮黏液黏附作用的研究
Adhesion of Vibrio fluvialis to the skin mucus of flounder, Paralichthys olivaceus
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摘要: 为了解河流弧菌对牙鲆表皮黏液的黏附特性,采用3H-TdR同位素示踪方法研究了细菌浓度、孵育时间、孵育温度、pH值、阳离子、碳水化合物等因子对河流弧菌黏附作用的影响。试验结果表明:河流弧菌能很好地黏附于牙鲆表皮黏液,黏附量随菌浓度升高而增大并符合饱和黏附动力学:y=417.89ln(x)+691.57(R2=0.986);在30℃下黏附量随着孵育时间的延长而增加,180min趋于饱和;孵育温度在4~30℃范围内黏附量随温度升高而增多,30℃时黏附量最大;pH值酸性较碱性黏附作用明显,pH为5时最强;NaCl浓度在0.5%~4.5%范围内,细菌黏附作用随着浓度升高而增强,较纯水存在显著性差异(P<0.05);Ca2+,Mg2+能显著加强河流弧菌的黏附作用,但Ca2+作用明显超过Mg2+;葡萄糖等8种碳水化合物能显著促进河流弧菌的黏附作用(P<0.05)。以上结果说明:河流弧菌对牙鲆表皮黏液的黏附量较大,这有利于进一步感染、致病;河流弧菌的黏附作用受温度、pH值、阳离子、碳水化合物等多种环境因子的影响,较高的水温有利于河流弧菌的黏附,有利于疾病的发生。Abstract: Bacterial adhesion to fish mucus is an important step in the initial stage of infection.It is important to underst and the characteristics involved in the adherence of bacteria to fish mucus so that preventive measures can be devised to reduce infection.In order to get a better understanding of adhesion characteristic of pathogenic Vibrio fluvialis to the skin mucus of Paralichthys olivaceus,the influences of bacterial density,incubation time,incubation temperature,pH,cation,and carbohydrate on the bacterial adhesion were investigated by using 3H-TdR as isotope tracer.The results showed that:the adhesive quantity of V.fluvialis increased with bacterial densities,a saturation kinetics hyperbola:y= 417.89ln(x)+691.57 (R2=0.986),was obtained by plotting adhered bacteria against the density of bacteria added;adhesive quantity of V.fluvialis increased with incubation times and reached equilibrium after incubation for 180 min;bacterial adhesion enhanced with the increase of incubation temperatures (from 4 to 30) and NaCl concentrations (from 0.5% to 4.5%);V.fluvialis exhibited stronger adhesion under sourish condition,strongest adhesion was recorded at pH 5;Ca2+ and Mg2+ obviously promoted the bacterial adhesion respectively,Ca2+ more efficiently;8 kinds of carbohydrates tested promoted the bacterial adhesion remarkably.The results indicated that:pathogenic V.fluvialis could adhere to the skin mucus of P.olivaceus facility,which would convenient for the subsequently infection;the bacterial adhesion was environmental factors dependent,warm water facilitated the bacterial adhesion.The results also indicated that the bacterial adhesion of V.fluvialis was governable,which might be useful for the vibriosis control in fish culture.
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Key words:
- Vibrio fluvialis /
- Paralichthys olivaceus /
- skin mucus /
- adhesion
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LEE J V,SHREAD P,FURNISS A L,et al.Taxonomy and description of Vibrio fluvialis sp.nov.(synonym group F vibrios,group EF6)[J].Journal of Applied Microbiology,1981,50(1):73-94. BIOSCA E G,AMARO C.Siderophores and related outer membrane proteins in Vibrio spp.which are potential pathogens of fish and shellfish[J].Journal of Fish Diseases,1991,14(3):249-263. 廖志强,纪荣兴,梅园园.福建省主要海水养殖鱼类疾病及其防治[J].福建水产,2003,15(4):30-35. MONTGOMERY M T,KIRCHMAN D L.Induction of chitin-binding proteins during the specific attachment of the marine bacterium Vibrio harveyi to chitin[J].Applied and Environmental Microbiology,1994,60(12):4284-4288. PICKERING A D.The distribution of mucous ceils in the epidermis of the brown trout Salmo trutta(L.)and the char Salvelinus al pinus(L.)[J].Journal of Fish Biology.1974,6(2):111-118. 刘旭银,胡朝锦,税蔚晰.动物免疫实用技术[M].重庆:重庆大学出版社,1994. ACORD J,MASKELL J,SEFTON A.A rapid microplate method for quantifying inhibition of bacterial adhesion to eukaryotic cells[J].Journal of Microbiological Methods,2005,60(1):55-62. CHABRILLON M,BORDAS M A,MORINIGO M C,et al.Kinetics of adhesion of Listonella anguillarum to the mucus of gilt-head seabream,and the implication of surface components[J].Aquaculture Research,2004,35(4):403-409. BORDAS M A,BALEBONA M C,ZORRILLA I,et al.Kinetics of adhesion of selected fish-pathogenic Vibrio strains of skin mucus of gilt-head sea bream(Sparusauratu L.)[J].Applied and Environmental Microbiology,1996:62(10):3650-3654. BALEBONA M C,MORINIGO M A,FARIS A,et al.Influence of salinity and pH on the adhesion of pathogenic Vibrio strains to Sparus aurata skin mucus[J].Aquaculture,1995,132(1/2):113-120. 鄢庆枇,陈强,邹文政,等.不同环境条件对溶藻弧菌粘附大黄鱼肠粘液的影响[J].水产学报,2006,30(2):254-259. 常建波,宫向红,孙逢贤,等.养殖牙鲆弧菌病病原菌初步研究[J].海洋水产研究,2001,22(1):37-41. 莫照兰,茅云翔,陈师勇,等.一株牙鲆皮肤溃烂症病原菌的鉴定[J].微生物学报,2002,42(3):263-269. 莫照兰,茅云翔,陈师勇,等.养殖牙鲆鱼苗腹水症病原苗的鉴定及系统发育学分析[J].海洋与湖沼,2003,34(2):131-141. 王斌,邵占涛,陈营,等.2种乳酸菌对牙鲆体表、消化道黏液黏附及对4种致病菌的黏附抑制性[J].中国微生态学杂志,2005,17(6):419-421,424. 鄢庆枇,邹文政,纪荣兴,等.应用荧光抗体技术检测牙鲆体内的河流弧菌[J],海洋科学.2006,30(4):16-19. BRADFORD M M.A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle ofprotein-dye binding[J].Analytical Biochemistry,1976,72(1/2):248-254. VESTERLUND S,PALTTA J,KARP M,et al.Measurement of bacterial adhesion-in vitro evaluation of different methods[J].Journal of Microbiological Methods,2005,60(2):225-233. LEE Y K,LIM C Y,TENG W L,et al.Quantitative approach in the study of adhesion of lactic acid bacteria to intestinal cells and their competition with enterobacteria[J].Applied and Environmental Microbiology,2000,66(9):3692-3697. BELAS M R,COLWELL R R.Adsorption kinetics of laterally and polarly flagellated Vibrio[J].Journal of Bacteriology,1982,151(3):1568-1580. MURRAY P R,BARON E J,PFALLER M A,et al.Manual of Clinical Microbiology[M].American Society for Microbiology,Washington,DC,1999:497-502. HOOD M A,WINTER P A.Attachment of Vibrio cholerae under various environmental conditions and to selected substrates[J].FEMS Microbiology Ecology,1997,22(3):215-223. 郭琨.海洋手册[M].北京:海洋出版社,1984:41. ATSUMI T,MCCARTER L,IMAE Y.Polar and lateral flagellar motors of marine Vibrio are driven by differention-motive forces[J].Nature,1992,355:182-184. WANG X H,LEUNG K Y.Biochemical characterization of different types of adherence of Vibrio species to fish epithelial cells[J].Microbiology,2000,146:989-998.
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