Carvacrol oil inhibits biofilm formation and exopolysaccharide production of Enterobacter cloacae

This study aimed to analyze the effect of carvacrol oil on biofilm formation of Enterobacter cloacae. The minimum inhibitory concentration of carvacrol oil against E. cloacae was 256 μg/mL. The biofilm formation was inhibited by carvacrol oil at sub-inhibitory concentrations of 64 and 128 μg/mL. The bacterial cell motilities and biofilm exopolysaccharide (EPS) production were both inhibited by carvacrol oil. Correlation analysis indicated that the inhibition of biofilms in mature stages was related to the decrease of EPS production. The biofilm thickness and excretion of extrapolymeric matrix were reduced by carvacrol oil, as shown by scanning electron microscopy and confocal laser scanning microscopy analyses. Transcriptional analysis showed that carvacrol oil down-regulated 24 of 29 selected genes, including curli fimbriae genes (csgABCEFG) and colonic acid polysaccharide genes (wcaABCDEFKLM, wza, wzb, wzc, gmd, manB, manC) which are required for biofilm formation of E. cloacae. In addition, different antimicrobial agents, including 1% lactic acid, 1% phenyllactic acid, and 1024 μg/mL of carvacrol oil considerably inactivated the bacterial cells in carvacrol-treated biofilms. Hence, carvacrol oil effectively inhibited biofilm formation and EPS production of E. cloacae, which is useful for further inactivating E. cloacae biofilms by other antimicrobial agents.