Synergistic bactericidal effect of clove oil and encapsulated atmospheric pressure plasma against Escherichia coli O157:H7 and Staphylococcus aureus and its mechanism of action

Abstract

We investigated the bactericidal effect of clove oil and encapsulated atmospheric pressure plasma (EAP), individually or in combination, against Escherichia coli O157:H7 and Staphylococcus aureus. The bactericidal effect of the combined treatment was also investigated in inoculated beef jerky. For both pathogens, clove oil and EAP single treatments resulted in less than 3.0-log reductions, whereas the combined treatment resulted in more than 7.5-log reductions. The disc-diffusion assay and gas chromatography-mass spectrometry showed no changes in both the clear zone diameter and chemical composition of clove oil before and after the EAP treatment. Significant changes in cell membrane permeability and cell morphology resulting from the combined treatment of clove oil and EAP were evidenced by increased in UV absorption of cell supernatants, increased cell staining with propidium iodide, and changes in cell structure revealed by transmission electron microscopy. The synergistic bactericidal effects of clove oil and EAP against both pathogens were also observed in inoculated beef jerky, but the treatments were less effective against S. aureus, presumably due to thicker peptidoglycan layer. Experiments also demonstrated that the synergistic bactericidal effects between clove oil and EAP are due to clove oil increasing the susceptibility of the bacteria to subsequent EAP treatment, and does not involve alteration of the antibacterial activity of clove oil by EAP.