This study assessed the occurrence of an enhancing inhibitory effect of the combined application of Origanum vulgare (OV) and Rosmarinus officinalis (RO) essential oils against bacteria associated to minimally processed vegetables using the determination of Fractional Inhibitory Concentration (FIC) index, kill-time assay in vegetal broth and application in vegetable matrices. Moreover, it was determined chemical composition of the essential oils and their effects alone and in mixture on sensory characteristics of minimally processed vegetables. Carvacrol (66.9 g/100 g) was the most prevalent compound in OV essential oil, while for RO was 1.8-cineole (32.2 g/100 g). OV and RO essential oil showed MIC in a range of 1.25-5 and 20-40 μL/mL, respectively. FIC indices of the combined application of the essential oils were 0.5 against Listeria monocytogenes, Yersinia enterocolitica and Aeromonas hydrophilla suggesting a synergic interaction. Only for Pseudomonas fluorescens FIC index was 0.75 purposing additive effect. Application of the essential oils alone (MIC) or in mixture (¼ MIC + ¼ MIC or ¼ MIC + ½ MIC) in vegetable broth caused significant decrease (p b 0.05) in bacterial count over 24 h. Mixture of essential oils reduced (p b 0.05) the inocula of all bacteria in vegetable broth and in experimentally inoculated fresh-cut vegetables. Similar efficacy was found to reduce the autochthonous microflora in vegetables. Sensory evaluation of vegetables sanitized with essential oils revealed that the scores of the most evaluated attributes fell between like slightly and neither like nor dislike. The combination of essential oils at sub-inhibitory concentrations could mean an interesting approach to sanitize minimally processed vegetables.

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This study evaluated the influence of the phenolic compounds carvacrol (CAR) and thymol (THY) on some physiological characteristics and on the modulation of the secretion of some staphylococcal virulence factors, that is, coagulase and enterotoxin. This study also investigated possible mechanisms for the establishment of the anti-staphylococcal activity of these compounds. Sublethal concentrations (0.3 and 0.15 μL/mL) of CAR and THY inhibited the activity of the enzymes coagulase and lipase and led to a decrease in salt tolerance. At the tested sublethal concentrations, both CAR and THY led to a total suppression of enterotoxin production. The loss of a 260-nm-absorbing material and an efflux of potassium ions occurred immediately after the addition of CAR and THY at 0.6 and 1.2 μL/mL and increased up to 120 min of exposure. Electron microscopy of cells exposed to CAR and THY (0.6 μL/mL) revealed that individual cells appeared to be deformed, with projections of cellular material. The observations of leakage of cellular material and an altered cell surface suggest that gross damage to a cell’s cytoplasmic membrane, which results in a disruption in protein secretion, could be responsible for the anti-staphylococcal properties of CAR and THY.

ABSTRACTOvernight exposure ofSalmonella entericaserovar Typhimurium to sublethal amounts ofOriganum vulgareessential oil (OV) and carvacrol (CAR) did not result in direct and cross-bacterial protection. Cells subcultured with increasing amounts of OV or CAR survived up to the MIC of either compound, revealing few significant changes in bacterial susceptibility.

This study evaluated the occurrence of an enhancing inhibitory effect of the combined application of Origanum vulgare L. essential oil and acetic acid against Staphylococcus aureus by the determination of Fractional Inhibitory Concentration (FIC) index and kill-time assay in nutrient broth, meat broth and in a food model (meat pieces). Acetic acid showed MIC and MFC of 0.6 and 1.25 µL.mL -1 , respectively. For O. vulgare essential oil MIC and MBC were 1.25 and 2.5 µL.mL -1 , respectively. FIC indexes of the mixture of essential oil and acetic acid at MIC x ½ were ≤ 1.0, showing an additive effect. No synergy was found at kill-time study. Antistaphylococcal effect of the antimicrobials alone or in mixture (MIC x ½) was lower in meat than in nutrient and meat broths. The effective combination of essential oils and organic acids could appear as an attractive alternative for the food industry, as the doses to inhibit the microbial growth in foods can be lowered.