## 4 Discussion  
_S. thymbra, T. capitatus_, and _O. hitrum_ EOs have been previously tested as potential antimicrobial and antibiofilm agents [3; 31; 29; 30; 31; 32]. In the present study, determining the Minimum Inhibitory Concentration (MIC) first allowed for a primary understanding of the antimicrobial efficacy of the EOs, WSTRTc extracts, and their main components. This step served as a precursor to the subsequent evaluation of their MBIC, enabling a comprehensive characterization of their antimicrobial activity. The MIC values indicated that the tested EOs, WSTRTc extracts, and their main components were effective antimicrobial agents against selected pathogenic bacteria. Notably, all main compounds tested (thymol, carvacrol, \(p\)-cymene, and \(\gamma\)-terpinene) exhibited low MIC values, underscoring their antimicrobial activity. These findings are consistent with previous studies demonstrating the efficacy of these compounds as natural antimicrobial agents [33; 34; 35; 36]. The present study showed that all EOs and their four main components had antibioticfilm activity against common pathogens. A variation between MIC and MBIC values was observed, highlighting the differences between inhibiting planktonic bacterial growth \begin{table}
\begin{tabular}{c c c c c c c c c} \hline \hline \multirow{2}{*}{**Bacterial Strains**} & \multicolumn{2}{c}{**Thymol**} & \multicolumn{2}{c}{**Carvacrol**} & \multicolumn{2}{c}{\(p\)**-Cymene**} & \multicolumn{2}{c}{\(\gamma\)**-Terpinene**} \\  & **MBIC** & **MBEC** & **MBIC** & **MBEC** & **MBIC** & **MBEC** & **MBIC** & **MBEC** \\ \hline _E. coli_ & 0.3 & 1.2 & 0.6 & 2.4 & 0.1 & 0.5 & 0.5 & 2.1 \\ _L. monocytogenes_ & 0.6 & 2.4 & 2.4 & 9.8 & 0.3 & 1.1 & 2.1 & 8.5 \\ _S. Enteritidis_ & 0.3 & 1.2 & 0.6 & 2.4 & 0.3 & 1.1 & 0.5 & 2.1 \\ _P. mirabilis_ & 0.6 & 2.4 & 1.2 & 4.9 & 0.3 & 1.1 & 2.1 & 8.5 \\ \hline \hline \end{tabular}
\end{table} 
Table 5: Minimum Biofilm Inhibition Concentration (MBIC) and Minimum Biofilm Fractionation Concentration (MBEC) (mg/mL) of the main components of the EOs (thymol, carvacrol, \(p\)-cymene, and \(\gamma\)-terpinene) against pathogenic bacteria. and preventing biofilm formation. MIC values represent the minimum concentration needed to inhibit the growth of planktonic bacteria, whereas MBIC values reflect the higher concentrations often required to interfere with biofilm formation, due to the enhanced resistance of biofilms. The observed differences underscored the resilience of biofilms, which derive protection from mechanisms, such as the production of an extracellular polymeric substance (EPS) matrix and the activation of quorum-sensing systems. Interestingly, in some cases, MBIC values were lower than MIC values, potentially due to stress-induced suppression of bacterial adhesion genes during the initial biofilm formation phase. This phenomenon suggested that the antimicrobial compounds tested may effectively disrupt early biofilm formation steps by interfering with adhesion mechanisms, as previously reported in the literature [37; 38; 39; 40]. Moreover, the observed variation between MIC and MBIC values highlighted the importance of strain-dependent and environmental factors in influencing bacterial adherence and biofilm formation. These factors include the structural properties of bacterial surface appendages, the composition of the EPS matrix, and the specific interactions between bacterial cells and their environment [41]. Many studies indicated the antimicrobial effect of _S.thymra_ EO [42; 43], but the antibiofilm activity was less examined. [PERSON] et al. [3] tested the ability of _S.thymra_ EO and its hydrosol to eliminate preformed monoculture and mixed culture biofilms of _L. monocytogenes_ compared to hydrochloric acid, lactic acid, and sodium hydroxide. The results suggested a substantial bactericidal effect of the oil and its hydrosol, but the only concentration examined was 1% (\(v/v\)). In the present study, the MBIC of _S.thymra_ EO against _L. monocytogenes_ was estimated at 0.6 (mg/mL).