## 3 Results
### Antibiofilm Activity  
The antibiofilm activity of the tested _G. hederaca_ extracts, expressed as a percentage of inhibition of biofilm formation or biofilm removal, is presented in Figure 2. Four strains of bacteria were used for this experiment. All tested extracts reduced the biofilm formation, with the highest ratio demonstrated by extract obtained at the 40 \({}^{\circ}\)C compared to the other temperature variants. The percentage inhibition of the biofilm formation by _E. coli_ and _E. faecalis_ exceeded 90%, whereas in the case of _P. aeruginosa_ and _B. subtilis_ it was almost 90% (88.6% and 87.9%, respectively). The extract obtained at 50 \({}^{\circ}\)C also had a strong inhibitory effect on biofilm formation, displaying the strongest inhibition of biofilm formed by _E. coli_ (88.2%). The percentage reduction of biofilm formation by other bacteria was lower, \begin{table}
\begin{tabular}{c c c c c} \hline \hline \multirow{2}{*}{**Microorganism**} & **MIC/** & \multicolumn{4}{c}{_Glechoma hederaca_ Extracts,} \\ \cline{3-5}  & **MPC** & \multicolumn{4}{c}{**Extraction Conditions:**} \\ \cline{3-5}  & **(mg/mL)** & **40 \({}^{\circ}\)C** & **50 \({}^{\circ}\)C** & **60 \({}^{\circ}\)C** \\ \hline \multirow{2}{*}{_F. graminearum_ KZF 1} & MIC & 2.5 & 2.5 & 2.5 \\  & MFC & 2.5 & 2.5 & 2.5 \\  & MIC & 5.0 & 5.0 & 5.0 \\  & MFC & 5.0 & 5.0 & 5.0 \\ _A. alternata_ KZF 13 & MFC & 5.0 & 5.0 & 5.0 \\  & MIC & 1.25 & 1.25 & 1.25 \\ _S. scleroorum_ KZF 23 & MFC & 1.25 & 1.25 & 1.25 \\  & MIC & 5.0 & \textgreater{}5.0 & \textgreater{}5.0 \\ _B. cinerea_ BPR 187 & MFC & 5.0 & \textgreater{}5.0 & \textgreater{}5.0 \\  & MIC & 1.25 & 2.5 & \textgreater{}5.0 \\  & MFC & 1.25 & \textgreater{}5.0 & \textgreater{}5.0 \\ \hline \hline \end{tabular}
\end{table} 
Table 4: Fungistatic activity of _G. hederaca_ extracts obtained via SC-CO\({}_{2}\) against tested indicator fungi. with the lowest percentage of biofilm inhibition by _B. subtilis_ (55.0%). In the case of the extract obtained at 60 \({}^{\circ}\)C, the best results in the reduction of biofilm formation were observed for _E. coli_ (80.7%) and _E. faecalis_ (71.5%), whereas the inhibitory efficiency was lower for _B. subtilis_ (60.5%) and _P. aeruginosa_ (38.7%). The tested extracts showed variable effects on the removal of mature biofilm. In contrast to the inhibition of biofilm formation, the best results in biofilm removal were observed after the application of extracts obtained at 60 \({}^{\circ}\)C, within the range of 55.3-75.9%, depending on the microorganism. Only a small percentage of biofilm was removed by extracts obtained at 40 \({}^{\circ}\)C and 50 \({}^{\circ}\)C. Depending on the extract and bacterial strain, the efficiency of biofilm destruction ranged from 1.5% to 34.7%, with no effect on the biofilm formed by _P. aeruginosa_. The antibiofilm activity of _G. hederacea_ has not been described in the literature; however, several studies have described the effect of plant extracts obtained via supercritical fluid extraction. [PERSON] et al. (2020) [65] reported that extracts of _Artemisia arbosscens_, _Artemisia abyssinia_, _Pulicaria jaudettii_ and _Pulicaria petiolaris_ were effective as anti-biofilm formation agents for all tested bacteria at 1/2 MIC. The highest inhibition rate of biofilm formation by the extracts was observed against _B. subtilis_, whereas the lowest inhibition ratio was noted on _K. pneumoniae_, _S. typhimurium_, _P. aeruginosa_ and _E. faecalis_. [PERSON] et al. (2021) [66] found  
Figure 2: Effect of _G. hederacea_ SC-CO\({}_{2}\) extracts on biofilm formation and removal: (**a**) _E coli_, (**b**) _E. faecalis_, (**c**) _P. aeruginosa_, (**d**) _B. subtilis_. Averages with different letters (A–C) for biofilm formation and (a–c) for biofilm removal are significantly different at \(p\) < 0.05.  
that green cardamom essential oil obtained via supercritical fluid extraction prevented the biofilm formation of _E. coli_ O157:H7 and _S. typhimurium_ JSG 1748.