Optimizing Antibacterial Essential Oil Blends from Helichrysum plicatum, Petroselinum crispum, and Origanum vulgare for Dairy Preservation: Mixture Design and In Silico Analysis

This study demonstrates the potential of <i>Helicrisum plicatum</i> (<i>H. plicatum</i>), <i>Petroselinum crispum</i> (<i>P. crispum</i>) and <i>Origanum vulgare</i> (<i>O. vulgare</i>) essential oils and their combination against four strains of two bacterial species, <i>Listeria monocytogenes</i> (<i>L. monocytogenes</i>) and <i>Escherichia coli</i> (<i>E. coli</i>), isolated from milk. GC/MS and GC/FID analyses of <i>H. plicatum</i> essential oil revealed α-pinene (27.61%), γ-curcumene (20.7%) and neryl acetate (5.88%) as the main compounds present in <i>H plicatum</i> essential oil. The main components of the essential oil of <i>P. crispum</i> were α-pinene (17.34%), 1,3,8-<i>p</i>-menthatriene (23.66%), β-phellandrene (10.41%) and myristicin (12.7%). In <i>O. vulgare</i> essential oil, carvacrol (58.3%), <i>o</i>-cymene (15.4%) and thymol (6.0%) were predominant compounds. Subsequently, an augmented simplex-centroid mixture design was applied to optimize the antibacterial efficacy of EO formulations against <i>L. monocytogenes</i> and <i>E. coli</i>. The simultaneous optimization all responses indicates that the optimal antimicrobial formulation was achieved using a ternary mixture of <i>H. plicatum</i>, <i>P. crispum</i>, and <i>O. vulgare</i> in a 16:16:68 (<i>v</i>/<i>v</i>/<i>v</i>) ratio. In parallel, an in silico study of four main essential oil components evaluated their drug-likeness, pharmacokinetic and toxicity properties, and binding to bacterial targets. These major compounds satisfy the main standards for drug-like molecules, with simulations pointing to good oral absorption, an acceptable safety profile, and stable binding to key enzymes in <i>E. coli</i> and <i>L. monocytogenes</i>, which supports their antibacterial potential. Overall, these combined experimental and computational results point to oregano, parsley, and immortelle essential oils as promising natural antimicrobial options for improving the preservation of dairy products.