Mycotoxins are secondary metabolites of some fungal species and represent important contaminants of food and feed. This study aimed to explore the biological control activity of Bacillus megaterium BM344-1 volatile organic compounds (VOCs) on the growth and mycotoxin production of single representatives of the toxigenic species Aspergillus flavus, Aspergillus carbonarius, Penicillium verrucosum, and Fusarium verticillioides. In vitro co-incubation experiments indicated the P. verrucosum isolate as the most sensitive one, with a growth inhibition ratio of 66.7%, followed by A. flavus (29.4%) and F. verticillioides (18.2%). Exposure of A. flavus, P. verrucosum, and F. verticillioides to BM344-1 VOCs resulted in complete inhibition of aflatoxins (AFB₁, AFG₁, and AFG₂), ochratoxin A, and fumonisin B₁ (FB₁) synthesis on artificial media, respectively. In vivo experiments on maize kernels showed 51% inhibition of fungal growth on ears simultaneously infected with A. flavus spores and exposed to BM344-1 volatiles. Likewise, AF synthesis by A. flavus was significantly (p < 0.05) inhibited (25.34 ± 6.72 μg/kg) by bacterial volatiles as compared to that in control maize ears (91.81 ± 29.10 μg/kg). Gas chromatography-tandem mass spectrometry-based analysis of headspace volatiles revealed hexadecanoic acid methyl ester (palmitic acid) and tetracosane as bioactive compounds in the BM344-1 volatilome. Bacterial volatiles have promising potential to control the growth and mycotoxin synthesis of toxigenic fungi and may present valuable aid in the efforts to warrant food and feed safety.

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