First, inocula of the two Aspergillus flavus strains were prepared. For this purpose, the strains were initially grown on malt extract agar (Scharlab S.L., Barcelona, Spain) at 25 °C for 10 days. A spore suspension of each strain was collected by adding 10 mL of sterile 0.05% (vol/vol) Tween 80 (Scharlab S.L.) to each mold plate and then rubbing the surface with a glass rod. The suspension formed was filtered through 2 layers of gauze. The concentration of each spore suspension was quantified using a Neubauer chamber and a microscope (Olympus CX 400, Tokyo, Japan) and adjusted to 106 spores/mL with sterile water.
For antifungal activity, potato dextrose agar (PDA; Oxoid, Madrid, Spain) plates were prepared and 100 μL of each essential oil was added and spread over the surface of the plate. After spreading, the plates were allowed to dry under sterile conditions around a flame.
For the assay, 10 μL of the spore suspension of each mold was inoculated in a central spot of the plate. The strains were inoculated separately. Sterile water was used as a negative control. Plates were incubated at 25 °C for 10 days.
For antifungal activity, potato dextrose agar (PDA; Oxoid, Madrid, Spain) plates were prepared and 100 μL of each essential oil was added and spread over the surface of the plate. After spreading, the plates were allowed to dry under sterile conditions around a flame.
For the assay, 10 μL of the spore suspension of each mold was inoculated in a central spot of the plate. The strains were inoculated separately. Sterile water was used as a negative control. Plates were incubated at 25 °C for 10 days.