## 4 Results  
In the first line of the investigation, we have scrutinized the E-, T-, and TT-EOs with regard to their content of antimicrobial substances. The GC-MS analysis confirmed the presence of specific active compounds within E-EO (including 1,8-cineole and \(\gamma\)--terpinene), T-EO (including thymol and \(p\)-cymene), and TT-EO (including \(\gamma\)--terpinene and terpinen-4-ol). The full list of active compounds identified is presented in Figure S1 and Table S1. Having proven that the tested EOs contain antimicrobial substances, we have analyzed the antimicrobial and antibiofilm activity of EOs' liquid fractions against microorganisms in planktonic and biofilm forms using the microdilution method. The rationale behind this preliminary research was the fact that the crucial EOs' components (such as thymol) display activity in both liquid and volatile fractions. Thus, the confirmation of antimicrobial activity of the liquid fraction would be a strong assumption for the potential activity of the volatile fraction. The results presented in Table S2 indicate a higher activity of all the tested liquid EOs against the applied microorganisms in planktonic than in biofilm form. Next, we analyzed the ability of _S. aureus_, _C. albicans_, and _P. aeruginosa_ strains to form biofilm (Figure 2) on the agar surface. A confirmation of the presence of a settled, multi-cellular biofilm formed on agar was a prerequisite condition for the performance of the core part of this research, namely AntiBioVol test for the E-, T-, and TT-EOs. The results of the performed AntiBioVol test suitability with the use of ethanol (which is a substance of known volatile antimicrobial activity) showed the correctness of the applied experimental setting--the number of biofilm-forming cells remaining after exposure to ethanol was lower than in the positive control of biofilm growth, regardless of the microorganism applied (Figure 3A-C). With regard to EOs, the results presented in Figure 3A-C show that none of the applied volatile fractions of EOs were able to completely eradicate bacterial or yeast biofilm, although the number of biofilm-forming cells after the exposure to all types of EOs was lower than in non-treated control settings. The volatile T-EO acted more efficiently against _S. aureus_ biofilm than TT- and E-EO, while TT- and T-EO was stronger against _P. aeruginosa_ biofilm than E-EO. Next, we have measured the activity of three concentrations of TT-EO against _S. aureus_ biofilm within a single measurement plate [Figure 4]. The comparison of TT-EO's antibofilm activity in a single vs. separate settings revealed a lack of statistical significance between them (K-W test \(p\) < 0.05, with post-hoc [PERSON]'s analysis) indicating no transmission of TT-EO volatile fractions between various plate wells within a single test plate. Similarly, no transmission of TT-EO solvent (PEG) was detected in the control setting (Figure S5). Finally, we have performed the conventional assessment of EOs' volatile fraction's antimicrobial activity referred to as the inverted Petri plate method to check whether such initial technical parameters as plate diameter, plate height, poured agar volume, and agar height translate into an outcome such as the size of the microbial growth's inhibition zone (Tables 1 and 2). Moreover, the reader can find a comparison of the inverted Petri dish method and the conventional disc diffusion method (which, being referred to as the aromatogram [26], is also applied for EOs' volatile fraction activity measurement) in Figures S6 and S7. As can be seen in Table 1, the differences between agar height obtained in Petri dishes of 15, 9, and 6 cm translate into differences in air volume between the agar surface and the lid of the dish. In the case of dishes 15 cm in diameter vs. dishes 9 and 6 cm in diameter, the aforementioned parameter differed by 75% and 83%, respectively. In turn, the difference in air volume between the 9 and 6 cm diameter dish was 31%.