The distribution and survival of the food-borne pathogen Vibrio parahaemolyticus is associated with its biofilm formation ability, which is affected by various environmental conditions.This study aimed to assess the effect of Ethylenediaminetetraacetic acid (EDTA) that influenced the development of biofilm formation by Vibrio parahaemolyticus.Attachment and biofilm development, inferred by the level of attached biomass, were measured in vitro using a colorimetric 96-well microtiter plate assay on the surface of biomaterials made from polyvinyl chloride (PVC).The results showed that adding 0.1 mM of EDTA had the strongly inhibit biomass in the early stage (5 h) of biofilm formation, while the addition of EDTA after 98 h had no significant inhibitory effects (p <0.05) , as 0.1 mM EDTA efficiently reduced the biofilm formation of Vibrio parahaemolyticus but not affected the planktonic growth.Under conditions of saturating EDTA with excess of cations, biomass did not significantly change (p <0.05) , demonstrating that the biofilm inhibition had no direct relationship with the chelating properties of EDTA.EDTA was shown to inhibit cell-to-surface interactions and cell-to-cell interactions, which at least partially contributed to the suppressed initial adherence.The study indicated that EDTA influenced biofilm process by inhibiting the initial adherence of Vibrio parahaemolyticus cells onto abiotic surfaces and may serve as an antibiofilm agent.