| Summary: | Natural antimicrobials have gained interest as possible inhibitors of biofilm formation. The aim of the present study was to determine the efficacy of antimicrobials derived from essential oils (carvacrol, thymol) plus bacteriocin AS-48 immobilized on two plastic supports (low density polyethylene and polyethylene⁻polyamide films) on bacterial inactivation. The polyethylene⁻polyamide vacuum-packaging plastic film activated with a combination of thymol plus enterocin AS-48 was the most effective in reducing the concentrations of viable planktonic and sessile cells for <i>Listeria innocua</i>, <i>Lactobacillus fructivorans</i>, <i>Bacillus coagulans</i>, and <i>Bacillus licheniformis</i>. Results from the study highlight the potential of polyethylene⁻polyamide film activated with thymol plus enterocin AS-48 for reducing the viable cell concentrations of spoilage Gram-positive bacteria and <i>Listeria</i> in both planktonic and sessile states.
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