致病性溶藻弧菌生物膜形成特性研究

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(P0.05);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(P0.01)。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。

Characteristics of Vibrio alginolyticus Biofilm Formation

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 remarkably higher OD590 values at pH 7~9,and peaked at pH 7.Addition of 1.5% CaCl2 significantly promoted the bacterial 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.