Antifungal and antitoxin effects of propolis and its nanoemulsion formulation against Aspergillus flavus isolated from human sputum and milk powder samples

Abstract Background and Aim: Aspergillus flavus causes human and animal diseases through either inhalation of fungal spores or ingestion of mycotoxins as aflatoxins produced in human and animal feed as secondary metabolites. This study was aimed to detect the incidence of A. flavus and its aflatoxins in human sputum and milk powder samples and explore the efficacy of pure propolis (PP) and propolis nanoemulsion (PNE) as natural decontaminants against fungal growth and its released aflatoxins. Materials and Methods: A. flavus was isolated by mycological culture and identified macroscopically and microscopically. Coconut agar medium and thin-layer chromatography (TLC) were used to qualitatively detect aflatoxins in the isolated strains. Toxins were extracted from toxigenic strains by the fast extraction technique. The quantitative detection of toxin types was explored by high-performance liquid chromatography (HPLC). PNE was prepared by a novel method using natural components and characterized by Fourier-transform infrared spectroscopy, Zetasizer, and transmission electron microscopy. The effects of PP and PNE on A. flavus growth and its toxin were determined by the well-diffusion method and HPLC. Results: The mycological culture showed that 30.9% and 29.2% of sputum and milk powder samples were positive for A. flavus, respectively. TLC confirmed the production of 61.8% and 63.2% aflatoxin by the isolated strains in sputum and milk powder, respectively. PP and PNE showed antifungal activity on A. flavus growth with mean±standard error (SE) inhibition zones of 27.55±3.98 and 39.133±5.32 mm, respectively. HPLC revealed positive contamination of toxin extracts with AFB1, AFB2, and AFG2 at 0.57±0.026, 0.28±0.043, and 0.1±0.05 mg/L, respectively. After treatment with PP and PNE, a significant decrease in AFB1, AFB2, and AFG2 concentrations was observed. Conclusion: This study suggested using propolis and its nanoformulation as antifungal and antitoxins in human medicine and the food industry to increase the food safety level and stop food spoilage.

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