## 2 Materials and Methods
### Biofilm Formation Ability  
Biofilm formation of _C. albicans_ ATCC 10231 and the effect of ATE on the suppression of biofilm formation were assessed using the standard microplate culture method. Microplates with 96 wells had a comfortable volume without pouring out--200 \(\mu\)L. YLF culture was grown in advance for 24 h at a temperature of 37 \({}^{\circ}\)C on the Sabouraud broth medium, in a volume of 10 mL, then centrifuged at 2.4 \(\times\) 10\({}^{3}\) rpm, and a fungal suspension was prepared. In our work, we used the 2.0 McFarland turbidity standard (HiMedia, Mumbai, Maharashtra, India) with a densitometer DEN-1B (Biosan, Riga, Latvia). In the wells of a sterile polystyrene plate 100 \(\mu\)L of the Heart brain broth (Hbb) medium was added. Next, 200 \(\mu\)L of previously prepared ATE was added in the second empty well of the first row. The first well was left as a control. By successive transfer of 100 \(\mu\)L of the solution from the second well to the third, from the third to the fourth \(\ldots\), etc., we reduced the concentration of ATE by half each time [25]. Further, _C. albicans_ inoculum was added in each well and cultivated 48 h at 37 \({}^{\circ}\)C. The microorganism culture was already in Hbb medium. This means that 75% of the volume of the well was always occupied by nutrition medium. And only 25% of the well was extracted in different concentrations. Nutrients were not depleted. They were always in the same quantity. The plate was cultured with the lid tightly closed. The progress of the experiment is presented in Table 1.  
After cultivation, the microtiter plate was washed three times with phosphate buffer (PB) and dried for 15 min. A 1% solution of crystal violet was added to each well. The plate was incubated for 15 min, washed again with PB solution, and dried for 15 min. After that, 150 \(\mu\)L of 96% ethanol was added to each well, then incubated for 15 min and the result was recorded on the microplate reader at a wavelength of 492 nm [25; 43; 44]. \begin{table}
\begin{tabular}{l l l l l l l l l l l l l l} \hline \hline  & **1** & **2** & **3** & **4** & **5** & **6** & **7** & **8** & **9** & **10** & **11** & **12** \\ \hline Action 1 & Hbb 100 \(\mu\)L & & & Hbb 100 \(\mu\)L & & Hbb 100 \(\mu\)L & & Hbb 100 \(\mu\)L & & Hbb 100 \(\mu\)L & & Hbb 100 \(\mu\)L & & Hbb 100 \(\mu\)L & & Hbb 100 \(\mu\)L \\ \hline Action 2 & & \multicolumn{4}{l}{+ATE} & & & & & & & & & & & \\  & 200 \(\mu\)L & & & & & & & & & & & & \\ \hline Action 3 & Not filtrated & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer & Transfer \\  & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L & 100 \(\mu\)L \\ \hline \multirow{2}{*}{
\begin{tabular}{l} Ther of ATE \\ no ATE \\ \end{tabular} } & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} & \multicolumn{4}{l}{+100 \(\mu\)L of} \\  & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} & \multicolumn{4}{l}{culture} \\ \hline Action 5 & Wait for 48 h & & & & & & & & & & & & \\ \hline \hline \end{tabular}
\end{table} 
Table 1: Stages of the study ATE effect on the _C. albicans_ biofilm formation. The average decrease was measured and used to calculate the biofilm inhibition percentage by ATE, where OD AS was the optical density average of _C.