## 2 Materials and Methods
### Extracts Characterisation and Antimicrobial Activity  
The chromatographic analyses were carried out at the Analytical Center of the Institute of Chemistry of the Federal Rural University of Rio de Janeiro. A 5890 Series II Gas Chromatograph (Agilent, USA) was used, equipped with a flame ionization detector, and an injector in \"split\" mode (1-20) was used to separate and detect the constituents of the _S.__terbeinthifolia_ extracts. The substances were separated on a fused silica capillary column (Fact Four VF-5 ms, 30 m \(\times\) 0.25 mm \(\times\) 0.25 \(\upmu\)m, Agilent J&W). Helium was used as a carrier gas with a flow of 1 mL min\({}^{-1}\). The oven temperature was programmed at 60 \({}^{\circ}\)C for 2 min, with an increment of 5 \({}^{\circ}\)C min\({}^{-1}\) to 110 \({}^{\circ}\)C, followed by an increment of 3 \({}^{\circ}\)C min\({}^{-1}\) to 150 \({}^{\circ}\)C and, finally, followed by an increment of 15 \({}^{\circ}\)C min\({}^{-1}\) to 290 \({}^{\circ}\)C, maintained for 15 min. The injector and detector temperatures were 220 and 290 \({}^{\circ}\)C, respectively.  
The gas chromatograph coupled to the QP-2010 Plus mass spectrometer (Shimadzu, Japan) was used to separate and analyze the substances present in the extract from the _S.__terbeinthifolia_ fruits. The helium flow, capillary column, and temperature programming for the GC-MS analysis were the same as described for the GC-DIC analysis. The injector and interface temperatures were 220 and 250 \({}^{\circ}\)C, respectively. The mass spectrum was obtained in a quadrupole detector operating at 70 eV, with a mass range between 40 and 400 \(m/z\) and a ratio of 0.5 scan s\({}^{-1}\).  
The identification of the extracts constituents was performed based on the comparison of the calculated retention indices [18] and the mass spectra obtained with those described in the literature [19] and in the equipment database (NIST Library 2008).  
The antimicrobial activity of the extracts was evaluated according to the National Committee for Clinical Laboratory Standards [20] and [PERSON] et al. [21] through the investigation of the MIC (minimum inhibitory concentration) against the strains of _Staphylococcus aureus_ (ATCC 6338) and _Escherichia coli_ (ATCC 8739). Bacterial samples, transferred from frozen stocks at \(-20\)\({}^{\circ}\)C, were seeded on brain heart infusion (BHI) agar and incubated in a bacteriological incubator (Quimis, Brazil) at 37 \({}^{\circ}\)C for 24 h, after which they were cultivated in BHI agar plates for inoculum preparation. The biomass was removed and suspended in a sterile NaCl solution (0.89%), homogenizing the bacterial suspensions until obtaining turbidity equivalent to the standard 0.5 of the McFarland scale (1.5 \(\times\) 10\({}^{8}\) CFU\(\cdot\)mL\({}^{-1}\)). Then, 30 \(\upmu\)L samples of bacterial suspensions were inoculated into 30 mL of BHI medium. For the MIC irrigation, the initial inoculum was 1-2\(\times\) 10\({}^{5}\) CFU\(\cdot\)mL\({}^{-1}\) and the oil concentrations ranged from 1600 to 3.125 \(\upmu\)g. mL\({}^{-1}\). The results were observed after the addition of 40 \(\upmu\)L of resazurin solution (100 \(\upmu\)g\(\cdot\)mL\({}^{-1}\)) with reincubation at 36 \({}^{\circ}\)C for 2 h. The blue color on  
\begin{table}
\begin{tabular}{c c c} \hline \hline
**Parameter** & **Value** & \\ \hline C & 51.3 & \\ H & 6.3 & \\ N & 1.2 & wt\% Dry Biomass \\ O & 41.0 & \\ S & 0.19 & \\ \hline Ash & 6.0 & \\ Volatile Matter & 64.3 & \\ Moisture & 14.4 & wt\% As Measured \\ Fixed Carbon & 15.3 & \\ \hline \hline \end{tabular}
\end{table}
Table 2: Average chemical characterization of all varieties of biomass.  
the microplates did not follow the growth of the investigated microorganisms, while the pink color showed bacterial growth.