Background: Aflatoxin B1 (AFB1), a potent group I carcinogen produced by Aspergillus species, poses a significant food safety challenge. This study evaluates the synergistic efficacy of heat-inactivated Lactobacillus(L.) brevis and L. paracasei (paraprobiotics) combined with ethanolic extracts of Punica (P.) granatum and Asparagus (A.) khorasanensis for AFB1 detoxification, extending prior findings on their anti-aflatoxigenic properties via gene expression modulation. Methods: AFB1 was produced using Aspergillus flavus PTCC 5006. Paraprobiotics were prepared by thermal inactivation, with cell wall integrity assessed via scanning electron microscopy (SEM) and Fourier-transform infrared spectroscopy (FTIR). Phytochemicals were characterized using high-performance liquid chromatography with diode-array detection (HPLC-DAD). AFB1 levels were quantified via HPLC with fluorescence detection (limit of detection LOD: 0.25 µg/mL; limit of quantification LOQ: 0.75 µg/mL). Dose-response relationships and synergy were evaluated using the Chou-Talalay combination index (CI). Results: Paraprobiotics reduced AFB1 by 47.2% (95% CI: 44.8–49.6%, Cohen’s d=1.8) at 108 CFU/mL, outperforming viable probiotics (30.6%, p<0.001). The binary extract mixture (250 µg/mL each) achieved a 43.7% reduction (95% CI: 40.5–46.9%, CI=0.78). The combined treatment yielded an 85.8% reduction (95% CI: 82.3–89.3%, CI=0.65, p<0.001), indicating strong synergy, correlated with downregulated aflatoxin biosynthesis genes (aflD, aflT, aflR, aflM). Conclusion: The paraprobiotic-phytochemical combination offers a dual-mechanism approach for AFB1 detoxification, integrating physical sequestration and transcriptional suppression. This strategy holds promise for food safety applications, warranting further in vivo and food matrix studies.
Kandi et al. (2026) studied this question.