Antibiofilm effect of warfarin on biofilm formation of Escherichia coli promoted by antimicrobial treatment

Enhancement of microbial biofilm formation by low antimicrobial doses is a critical problem in the medical field. The objective of this study was to propose a new drug candidate against the biofilm formation promoted by subinhibitory dose of antimicrobials. To determine the effect on biofilm formation of Escherichia coli, a subinhibitory concentration of lactoferrin (LF), a milk protein involved in a broad range of biological properties including antimicrobial action, or ampicillin (AMP), a typical antibiotic, was added to an E. coli cell culture in a 96-well microtiter plate. On the other hand, warfarin (WARF), an oral anticoagulant, or polymyxin B (PMB), a strong antibiotic for biofilm treatment, was added as an antagonist against the biofilm promoted by LF or AMP. The amount of biofilm formed at 100 μg/mL LF in lysogeny broth medium was four times higher than in the absence of LF. Meanwhile, it was found that WARF suppressed the LF-promoted biofilm formation to a level comparable with the LF-free condition. WARF worked in a similar manner to PMB, which is known as an antibiofilm agent. Furthermore, WARF could also suppress the biofilm promoted by AMP. In conclusion, this study suggests that WARF can work as an antibiofilm agent against the biofilm formation promoted by subinhibitory dose of antimicrobials.