Biofilms play an important role in the antibiotic resistance of encased bacteria, and their formation was regulated by quorum sensing (QS). Inhibiting the QS system may therefore degrade the integrity of a biofilm and expose the bacterial pathogens within it to the deleterious effects of molecules such as antibiotics. Moreover, the use of QS inhibitors (QSIs) may provide a novel approach for treating bacterial infections of aquacultures. In the present study, the bacterium Proteus mirabilis was identified as a potential producer of QSIs. Varying concentrations (0.1-1.1%) of filtrates prepared from the culture of P. mirabilis inhibited biofilm formation by the pathogens Pseudomonas aeruginosa, Vibrio harveyi, and Staphylococcus aureus by as much as 58.9%, 41.5%, and 41.9%, respectively. These filtrates as well as the crude aqueous extracts prepared from them increased the sensitivities of pathogens to the inhibitory effects of kanamycin. The filtrates also showed pathogenicity attenuation potential in P. aeruginosa by decreasing the production of virulence factors. Moreover, the filtrates did not influence the planktonic growth of these pathogens. The results indicate that P. mirabilis may act as a nonspecific (or broad-spectrum) inhibitor of biofilm formation that will help control infectious diseases that adversely affect the aquaculture industry.