副溶血性弧菌-霍乱弧菌混合生物被膜形成过程研究

Dynamic biofilm forming process of Vibrio parahaemolyticus and Vibrio cholera

摘要:

[目的]研究副溶血性弧菌（Vibrio parahaemolyticus，VP）和霍乱弧菌（Vibrio cholera，VC）混合生物被膜的形成过程。[方法]在4、8、12、24、36、48、60、72 h测定单独条件下VP、VC及其混合后生物被膜的形成情况，通过结晶紫染色法、平板菌落计数法、测定胞外多糖、胞外蛋白，通过荧光原位杂交（FISH）观察混合生物被膜形成。[结果]虽然形成的混合生物被膜量介于VC和VP之间，但混合生物被膜在形成过程中，成熟期后生物被膜量的变化较小，对环境的抗性增强。混合生物被膜中拥有更多的活菌，混合生物被膜形成过程中胞外蛋白和胞外多糖的变化体现出其可能在对抵御不适应环境中起重要作用，通过FISH可观察到不同时期生物被膜的变化过程。[结论]VC与VP共同形成生物被膜的过程中，混合生物被膜总量虽然减少，但混合生物被膜中拥有更多的活菌，这可能引起更大的危害。研究混合生物被膜形成过程中被膜的变化，可为有害生物被膜的控制提供基础。

Abstract:

[Objective] We studied the formation of a mixed biofilm formed by Vibrio parahaemolyticus and Vibrio cholerae. [Methods] Biofilm formation was quantified using crystal violet staining, abundance of extracellular polysaccharide and extracellular protein, fluorescence in situ hybridization and colony plate count. Biofilms formed by individual and mixed strains were determined from the start and the following intervals at 4, 8, 12, 24, 36, 48, 60 and 72 h. [Results] The amount of biofilm produced by the mixed strains was less than the total amount of biofilm produced by the individual strains, indicating there was no synergism between the strains. However, in the mixed biofilm the time of maturation was increased, the stability of the biofilm to external perturbation was enhanced and the cell density was higher. [Conclusion] The biofilm formed by a mixed population of V. parahaemolyticus and V. cholera is therefore more persistent and can potentially pose a greater risk for recontamination and subsequent development of disease.