Effects of phytogenic substances on growth and biofilm formation of Escherichia coli and Salmonella field isolates

The aim of this study was to investigate the effects of garlic oil, cinnamaldehyde, carvacrol, thymol, and thyme oil on growth and biofilm formation of Escherichia coli and Salmonella serotypes, including field isolates from livestock animals. Minimum inhibitory concentrations (MIC) were determined using broth micro-dilution method. Biofilm biomass was assessed by measuring the attached biomass with microtiter plate assay and crystal violet (CV) staining. The strongest antimicrobial effects on E. coli serotypes were observed for thymol at 150 ppm, followed by carvacrol and cinnamaldehyde at 300 ppm and thyme oil at 600 ppm. Similar results were obtained with Salmonella serotypes except for carvacrol (MIC value at 150 ppm). Garlic oil showed no growth inhibition on serotypes of E. coli and Salmonella up to 10000 ppm. Cinnamaldehyde proved to be the most effective substance in reducing E. coli CV-biofilm formation at sub-MIC level with a threshold concentration of 5 ppm, followed by carvacrol, thymol, and thyme oil at 40 ppm and garlic oil at 10000 ppm. CV-biofilm formation of Salmonella serotypes at sub-MIC level was clearly reduced with 40 ppm cinnamaldehyde and 80 ppm carvacrol, thymol, and thyme oil. No reduction of CV-biofilm formation was observed with garlic oil. The present study demonstrates a strong antibacterial activity of cinnamaldehyde, carvacrol, thymol, and thyme oil. Similar response of field isolates and type strains to these phytogenics suggests a general effect within the bacterial species tested. All four substances were also able to reduce CV-biofilm formation at sub-MIC level. Investigating phytogenics with bacterial field isolates contributes to the development of feed additives as alternatives to antibiotics in animal feed to increase productivity and animal welfare in modern livestock production.