Listeria monocytogenes, a bacterium that is transmitted by tainted food, causes the infection listeriosis. In this study, quercetin was tested for its antibacterial properties and effectiveness as a food additive in preventing the growth ofL. monocytogenescocktail (ATCC19117, ATCC19113, and ATCC15313) biofilms on crabs and shrimps. Quercetin showed the least bactericidal activity and no discernible microbial growth at a minimum inhibitory concentration (MIC) of 250 µg/mL. The biofilm inhibition was performed at sub-MICs (1/2, 1/4, and 1/8 MIC). There was no quercetin added to the control group. Additionally, the present work examines the expression of various genes related to biofilm formation and quorum sensing (flaA, fbp, agrA, hlyA, and prfA). The levels of target genes were all significantly down-regulated. Quercetin (0–125 µg/mL) on the surfaces of the crab and shrimp was studied; its inhibitory effects were measured as log reductions at 0.39–2.31 log CFU/cm2and 0.42–2.36 log CFU/cm2, respectively (p< 0.05). Quercetin reduced the formation of biofilms by disrupting cell-to-cell connections and causing cell lysis, which led to the deformation of the cells, evidenced by FE-SEM (field-emission scanning electron microscopy). These findings emphasize the significance of using natural food agents to target bacteria throughout the entire food production process.