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.

Artisanal vegetable fermentations are regaining popularity in industrialized countries, but they could be prone to contamination with foodborne pathogens. By simulating home or small-scale restaurant fermentations, we evaluated the microbiological safety of spontaneous carrot juice fermentations. Raw carrot juice was spiked with Listeria monocytogenes, Salmonella enterica subsp. enterica Typhimurium and Escherichia coli O157:H7, and the microbial dynamics were followed throughout the entire fermentation process by cultivation and amplicon sequencing. In addition, the behavior of these pathogens was also monitored after addition of raw cucumber juice and storage under refrigerated conditions to mimic post-contamination issues. Although the numbers of the pathogens increased during the first phase of the fermentation, the pathogens were not able to persist throughout the fermentation. Their numbers fell below the detection limit after 8 days of fermentation at 20 degrees C. Further investigation using amplicon sequencing also showed that there was no major impact on the general microbial dynamics of the spontaneous carrot juice fermentation. This indicates that the artisanal carrot juice fermentation is a robust process which resists the persistence of pathogens. More caution is needed however when mixing the final fermented product with a raw juice. When simulating pathogen post-contamination, both Salmonella enterica and Escherichia coli were able to survive in the refrigerated fermented juice up to 10 days after the fermentation. Listeria monocytogenes was detected up to 8 days in the refrigerated juice. Pasteurization of the raw juice before adding it to the fermented product is thus recommended.