Linalool as a potential agent for inhibiting Escherichia coli biofilm formation and exopolysaccharide production

Escherichia coli (E. coli) is one of the most common pathogens causing endometritis in dairy cows. The presence of genes encoding extended-spectrum β-lactamase (ESBL) and biofilm formation are important factors contributing to bacterial resistance, which poses a significant challenge to the treatment of endometritis in dairy cows. Essential oils containing linalool have been shown to improve the cure rate of bovine endometritis, but whether linalool can inhibit E. coli biofilm has not yet been reported. We proposed to ascertain the linalool implications on the development of E. coli biofilm and its extracellular polysaccharides, as well as to assess the impacts of linalool on E. coli in both planktonic and biofilm states. We discovered that the minimum biofilm inhibitory concentrations (MBICs) of linalool against E. coli were twice as high as the minimum inhibitory concentrations. Linalool exhibited a strong bactericidal effect on clinical E. coli strain producing ESBL and forming strong biofilm, regardless of whether they were in a planktonic or biofilm condition. Linalool suppressed the biofilm development in a way that was dependent on the dosage, with an MBIC 4 µL/mL. This was verified by the use of crystal violet test and scanning electron microscopy. Moreover, the CCK-8 assay and confocal laser scanning microscopy (CLSM) manifested significant reductions in live bacteria within the biofilm. The concentrations of extracellular polymeric compounds in the E. coli biofilm were also reduced. Furthermore, CLSM and RT-qPCR analysis confirmed that linalool (2 µL/mL) significantly suppressed exopolysaccharide (EPS) and the pgaABCD gene expression, regulating an essential exopolysaccharide expression in biofilm formation. These findings revealed that linalool effectively suppressed viable bacteria, EPS production, and E. coli biofilm formation, providing a theoretical foundation for alternative antibiotic therapy in endometritis in dairy cows and as a potential agent for preventing E. coli biofilm-related infections.