In this study， the characteristics of biofilm formation of single and mixed strains of Vibrio parahaemolyticus with different virulence genotypes from freshwater products under pure culture conditions and bile salts stress were investigated. The amount of biofilm formation and cell viability were detected by crystal violet staining and CCK-8 method， and the structure of biofilm was visually observed by confocal laser scanning microscopy （CLSM）. Bicinchoninic acid （BCA） method and phenol-sulfuric acid method were used to quantitatively analyze the contents of extracellular protein and extracellular polysaccharide before and after bile salts treatment. The results showed that bile salts treatment inhibited the formation of V. parahaemolyticus biofilm， significantly decreased the amount of biofilm formation and cell viability， and affected the degree of aggregation and disorganization of biofilm system， and disrupted the biofilm structure， while affecting the synthesis and secretion of extracellular protein and extracellular polysaccharide. Strains containing tdh gene can form more and more stable biofilms， and the mixed strains showed greater adaptability to bile salts stimulation and were able to respond rapidly to counteract this stress， thus ensuring more stable survival of the strains. This study provides a new theoretical support for the formation and control of V. parahaemolyticus mixed biofilms， which is of great significance for exploring the impact of V. parahaemolyticus carriage in freshwater products on food safety and human health， and provides an important reference for further research on its adaptive mechanism and prevention and control in the human gut.