Vibrio parahaemolyticus biofilm formation in the food factory environment was studied, in order to prevent and control the biofilm pollution effectively. The different vibration modes of food processing equipment (such as horizontal rotary vibration, bevel vibration and vertical rotary vibration) were simulated to investigate Vibrio parahaemolyticus formation process on the surfaces of glass and stainless steel for 72 h, and the effect of vibration on the biomass, architecture and extracellular polymeric substances of biofilm was analyzed. The results showed that:the biofilm formation under shaking cultivation decreased significantly; the biomass of biofilm was the least under vertical rotary vibration; the biomass of biofilm on the stainless steel surface is greater than that on glass surface under same shaking cultivation; the biofilm formation was reduced with the increasing of horizontal rotation speed; the vibration caused the biovolume decreased, and the porosity and homogeneity increased, the architecture of biofilm dispersed; and the amount of extracellular polysaccharides and extracellular proteins of the biofilm decreased. All results show that different vibration modes have different effects on the biofilm; the selection of the vertical rotary vibration mode out of the three vibration modes can reduce and inhibit the growth of the biofilm effectively; the vibration cause the reduction of exopolysaccharide and protein, affecting the structural characteristics such as porosity and homogeneity; and the structure and formation of biofilm becomes simple and decrease.