Vibrio parahaemolyticus, the causative agent of seafood-borne acute gastroenteritis worldwide, benefits from a sessile biofilm lifestyle that enhances survival outside the host, contributes to host colonization and infectivity but also leads to cross-contamination between processing equipment and foods. Insight into the regulatory circuit underlying biofilm formation may propose targeted strategies to interfere with a process that renders this bacterium remarkably adaptable to changing environments. Here, we mainly demonstrated the regulatory of two mobility structures (flagella, pili), extracellular matrix polysaccharides, the second messenger c-di-GMP and quorum-sensing system that contribute to biofilm formation.