The extracts were redissolved in 5% dimethyl sulfoxide (DMSO) to a concentration of 10 mg/mL and further diluted. The microdilution method [58 (link)] was performed to assess the antimicrobial activity against the Gram-negative bacteria Escherichia coli (ATCC 25922), Salmonella typhimurium (ATCC 13311) and Enterobacter cloacae (ATCC 35030), and Gram-positive bacteria: Staphylococcus aureus (ATCC 11632), Bacillus cereus (clinical isolate), Listeria monocytogenes (NCTC 7973). For antifungal assays, six micromycetes were used: Aspergillus fumigatus (human isolate), Aspergillus niger (ATCC 6275), Aspergillus versicolor (ATCC11730), Penicillium funiculosum (ATCC 36839), Trichoderma viride (IAM 5061) and Penicillium verrucosum var. cyclopium (food isolate). The minimum extract concentrations that completely inhibited bacterial growth (MICs) were determined by a colorimetric microbial viability assay, and minimum bactericidal concentration (MBC) and minimum fungicidal concentration (MFC) were also calculated. Streptomycin, ampicillin, ketoconazole, and bifonazole (Sigma-Aldrich, St. Louis, MO, USA) were used as positive controls, and 5% DMSO was used as a negative control.
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