Tween 80

Aspergillus parasiticus NRRL 2999 (a strain with the ability to produce mycotoxins such as AFB₁, AFB₂, AFG₁, and AFG₂) was kindly donated by the United States Department of Agriculture. This strain was isolated from soil in Uganda as reported by Singh and Cotty (2019) (link). The strain was prepared according to Wigmann et al. (2016) (link) by inoculating it in Malt Extract Agar (MEA; Acumedia, Lansing, Michigan, United States; Malt Extract: 20.0 g; Peptone: 1.0 g; Glucose: 30.0 g; Agar: 20 g; and Distilled Water: 1 L) and incubating it aerobically at 25°C in a BOD incubator for 5 days, at static condition in the dark. Conidia were collected by scraping the mycelium from each plate with sterile distilled water containing 0.1% Tween 80 (Labsynth, Diadema, Brazil). Subsequently, they were filtered and then centrifuged at 11,000 g three consecutive times for 15 min at 4°C (Sorvall Legend XTR, Thermo Scientific, Hampton, Unites States). The final concentration of conidia in the fungal suspension was determined using a Neubauer chamber (Sigma-Aldrich, Darmstadt, Germany). The conidial suspension was diluted and used to reach a final concentration of 5.0 log₁₀ cfu ml⁻¹ in the growth trials (checked by plate count).

The chemicals used in the assay were as follows: Tween^® 80 (T1029, Labsynth, Diadema, Brazil); Liovac^® 1112 [saturated fatty acid derived from hydrogenated castor oil, containing 88% pure 12-hydroxystearic acid (12-HSA), 12% oleic acid (C18), and palmitic acid (C16)] (Miracema-Nuodex, Campinas, Brazil); resveratrol (98% purity; Bothânica, Araraquara, Brazil); alamarBlue^TM, a cell viability reagent with resazurin (DAL1025, Thermo Fisher Scientific, Waltham, MA, USA); trypan blue solution (0.4%; 15250061; Gibco^TM, Waltham, MA, USA); DMEM (Dulbecco’s modified Eagle’s medium/high modified) (51441C; Merck, Darmstadt, Germany); fetal bovine serum (Gibco^TM); trypsin-EDTA (ethylenediaminetetraacetic acid) solution (T4049, Sigma-Aldrich, St. Louis, MO, USA); DMSO (dimethyl sulfoxide) (≥99.5% purity, D4540, Sigma-Aldrich); methyl methanesulfonate (99% purity, 129925, Sigma-Aldrich).