## 2 Materials and Methods
### Scale-Up and Optimization of the EO Treatment in GAC  
The treatment in Petri dishes (head space 63.62 cm\({}^{3}\)) with 10 \(\mu\)L thym EO (calculated vapor phase concentration 1.57\(\cdot\)10\({}^{-4}\)) was scaled up in glass airtight containers (head space 2650 cm\({}^{3}\)) (see Table 1). The concentration of the EO in the free space may be simply calculated dividing the total EO volume by the volume of the free space assuming that the whole amount of EO is vaporized during the test. This hypothesis may be verified considering the vapor pressure of the main EO components. At 25 \({}^{\circ}\)C, the vapor pressure of the pure components are the followings: 4.47\(\cdot\)10\({}^{-5}\) bar for Thymol, 4.77\(\cdot\)10\({}^{-4}\) bar for \(p\)-Cymene, 2.57\(\cdot\)10\({}^{-5}\) bar for \(\alpha\)-Terpinene and 7.432\(\cdot\)10\({}^{-5}\) bar for \(\gamma\)-Terpinene [26; 27; 28; 29]. Assuming an ideal behavior, it is possible to calculate the vapor pressure of each component in the solution equal to the product of the pure component vapor pressure and its molar fraction and this value results higher than the EO partial pressure in the free volume during the vaporization process. The containers are characterized by a shrinkage (see Figure 11) which allows to fit a plastic grid for Petri dish location. When 9 cm \(\O\) Petri dishes were used, each GAC contained one dish, and both the control and EO treated samples were triplicated. Using 5.5 cm \(\O\) Petri dishes it was possible to insert the three replicates (for the control and for the EO treated samples) in the same GAC. The container size allowed to carry out the EO dispersion in three different modalities. In the first one, the EO, 210 or 420 \(\mu\)L (calculated vapor phase concentrations 0.79\(\cdot\)10\({}^{-4}\) and 1. 58\(\cdot\)10\({}^{-4}\), respectively), was poured into a small glass beaker placed on the plastic grid. This experiment was carried out utilizing 90 mm diameter MEA Petri dishes, without the lid, inoculating \(A\). _alternata_ in the middle of the dish by means of a mycelial plug (\(\O\) 8 mm). A control glass airtight container was set-up without EO, and all the containers were maintained at 25 \({}^{\circ}\)C (see Figure 11A). In the second group of experiments the EO dispersion was realized by sticking a filter paper disk, 30 mm \(\O\), by means of bi-adhesive tape, on the top lid of the container; the disk was then soaked with 210 \(\mu\)L of EO. One or three Petri dishes, inoculated with \(A\). _alternata_ (inoculum age of 72 h), were positioned in GAC before the beginning of the treatment (see Figure 11B). In the third set of experiments each glass container was saturated with the EO vapors using a waterless dispersion device (see Figure 11C); in this case 10 \(\mu\)L of EO (calculated vapor phase concentrations 0.377\(\cdot\)10\({}^{-5}\)) were poured onto a non-woven fabric (TNT) gauze fixed inside the device and the dispersion was accelerated by the forced convection of air, result of a fan placed behind the gauze adjustment. During the saturation, the switched-on device was placed inside the closed glass container. After 30 min of dispersion, the device was quickly removed and Petri dishes with _A. alternata_ (inoculum age of 24, 72, and 96 h) were immediately placed onto the plastic grid. These three methods aimed to get different oil dispersion rates: in the first one the rate of dispersion in the container was the lowest, while in the second it was accelerated thanks to the higher contact area between the oil and the environment, being the highest rate that obtained with the third method, thanks to the turbulence obtained through the fan. This set of experiments was carried out utilizing 55 mm diameter \(\text{CZ}_{\text{mod}}\) Petri dishes. CZ has been chosen as synthetic medium to have the possibility to replace the carbon source (i.e., paper sheet instead of glucose). In this case, _A.