作者简介：姚刚(1983-)，男，硕士研究生；研究方向：环境微生物学.E-mail:[email protected] ,通讯作者：鄢庆枇(1963-),男,
教授；研究方向：水产病害．E-mail:[email protected][中文文摘]采用改良的微孔板法研究静置培养条件下致病性溶藻弧菌ND-01在酶标板上成膜情况。结果显示,溶藻弧菌的生物膜的OD590在16h达到峰值后,随培养时间延长出现下降;在102～108cfu/mL范围内,不同的初始菌密度对生物膜的OD590影响并不显著;NaCl质量分数为4.5%时,所形成生物膜的OD590最高;30℃,溶藻弧菌的生物膜OD590显著高于其他温度组(P<0.05);在胰蛋白胨大豆肉汤培养液中加入质量分数1.5%的CaCl2,能够显著促进生物膜形成,而加入MgCl2对于生物膜的影响则不显著;经过大黄鱼表皮黏液包被后,96孔板中生物膜的OD590显著高于其他部位组织提取液包被组(P<0.01)。致病性溶藻弧菌ND-01能够在聚苯乙烯材料的96孔酶标板表面形成稳定而明显的生物膜,且其生物膜的形成与多种环境因素有着密切关系。[英文文摘]A modified microtiter-plate test was introduced to determine biofilm formation of pathogenic Vibrio alginolyticus ND-01 on the surface of the polystyrene 96-well microtiter plate.Results showed that OD590 of V.alginolyticus strain ND-01 biofilm was found to peak after 16 h;within the 102~108cfu/mL,bacterial concentrations of V.alginolyticus exhibited no significantly effect on the biofilm formation;that the OD590 values of bacterial biofilm increased with the incubation temperatures of 4~30 ℃ and peaked at 30℃(P＜0.05); and that the OD590 values of bacterial biofilm fluctuated with different NaCl concentrations and optimized at 1% to 7%,the remarkablly higher OD590 values at pH 7-9,and peaked at pH7.Addition of 1.5% CaCl2 signficantly promoted the bacerial biofilm formation while MgCl2 played no significantly role:The OD590 of large yellow croaker the biofilm developed on the surface of the microtiter plate coated by the mucus layer on the skin of large yellow croaker was significant (P＜0.01).The results showed that V.alginolyticus strain ND-01 developed stable and evident biofilm,the biofilm formation was remarkably affected by environmental factors,such as temperature ,pH,NaCl concentration,and Ca2+ level.The results also showed that the bacterial biofilm formation of V.alginolyticus was governable,which would be useful for the epidemic control in fish culture.国家“863”计划项目(2007AA09Z115); 农业部公益性行业科研专项(200903029
