2.5.2. Minimum Inhibition Concentration (MIC) Tests

The MIC serves as the lowest concentration of the antimicrobial agent at which the visible growth of the microorganism is inhibited, primarily indicating the bacteriostatic effect of the agents without specific data on the microbial population condition. Pure cultures of Listeria monocytogenes (LM) and Staphylococcus aureus (SA) were cultivated in Müeller–Hinton broth to concentrations approximating 10⁻⁶ CFU/mL. Four types of antimicrobial agents were evaluated: MC, MCC, NC, and FC. These agents were tested at multiple concentrations, including 1000, 500, 250, 125, 62.5, 31.25, and 15.625 μg/mL [28]. Utilizing the Macro dilution method, serial decimal dilutions of the microbial cultures were conducted, and the agents were added at each of these concentrations for evaluation. Following thorough mixing via a vortex mixer, the tubes were incubated at 37 °C for 24 h. Control tubes, which contained only microbial cultures without any antimicrobial agents, were also maintained and assessed at a concentration of 250 μg/mL. Turbidity in the tubes was observed as an indicator of microbial growth, and additional validation was performed through culture and colony counting techniques. All tests were conducted in triplicate to ensure the reliability and repeatability of the observed results.