Antifungal activity of 2-ethylhexanol against Aspergillus ochraceus, related mechanisms, and grain fumigation verification

As an edible flavoring agent, 2-ethylhexanol (2-EH) has been reported for its broad-spectrum antifungal activity in recent years, highlighting its potential as an alternative to conventional chemical antifungal agents. However, its efficacy against Aspergillus ochraceus and the underlying mechanisms remain unclear. This study evaluated the inhibitory effects of 2-EH on A. ochraceus , elucidated its action mechanisms, and conducted grain fumigation experiments to validate its practical efficacy. The results demonstrated that 2-EH effectively inhibited A. ochraceus spore germination, hyphal growth, sporulation,and ochratoxin A (OTA) production in vitro, Treatment of 2-EH led to spore surface depression and fragmentation, cytoplasmic leakage, and organelle destruction, accompanied by increased extracellular conductivity and intracellular reactive oxygen species levels. Transcriptional analysis revealed that 2-EH downregulated key OTA biosynthetic genes ( pks , nrps , bzip , and hal ), global regulatory factors ( laeA and veA ), and ergosterol biosynthesis genes ( Erg2 and Erg11 ). Grain fumigation experiments confirmed that 2-EH effectively reduced total mold count, free fatty acid content, and OTA levels in corn, wheat, peanuts, and unhulled rice. These findings provide preliminary evidence supporting 2-EH's potential as an effective fumigant for grain storage. • A. ochraceus growth and OTA production was controlled by 2-ethylhexanol in vitro • The treated A. ochraceus cell membrane leakage and intracellular ROS were increased • OTA and ergosterol synthesis genes of treated A. ochraceus downregulated • Mold counts, OTA and FFA level of treated grains were effectively decreased