| Summary: | This study evaluated the efficacy of potentially probiotic fruit-derived <i>Lactobacillus</i> isolates, namely, <i>L. paracasei</i> 108, <i>L. plantarum</i> 49, and <i>L. fermentum</i> 111, to remove aflatoxin M<sub>1</sub> (AFM<sub>1</sub>) from a phosphate buffer solution (PBS; spiked with 0.15 µg/mL AFM<sub>1</sub>). The efficacy of examined isolates (approximately 10<sup>9</sup> cfu/mL) as viable and non-viable cells (heat-killed; 100 °C, 1 h) to remove AFM<sub>1</sub> was measured after 1 and 24 h at 37 °C. The recovery of AFM<sub>1</sub> bound to bacterial cells after washing with PBS was also evaluated. Levels of AFM<sub>1</sub> in PBS were measured with high-performance liquid chromatography. Viable and non-viable cells of all examined isolates were capable of removing AFM<sub>1</sub> in PBS with removal percentage values in the range of 73.9–80.0% and 72.9–78.7%, respectively. Viable and non-viable cells of all examined <i>Lactobacillus</i> isolates had similar abilities to remove AFM<sub>1</sub>. Only <i>L. paracasei</i> 108 showed higher values of AFM<sub>1</sub> removal after 24 h for both viable and non-viable cells. Percentage values of recovered AFM<sub>1</sub> from viable and non-viable cells after washing were in the range of 13.4–60.6% and 10.9–47.9%, respectively. <i>L. plantarum</i> 49 showed the highest AFM<sub>1</sub> retention capacity after washing. <i>L. paracasei</i> 108, <i>L. plantarum</i> 49, and <i>L. fermentum</i> 111 could have potential application to reduce AFM<sub>1</sub> to safe levels in foods and feeds. The cell viability of examined isolates was not a pre-requisite for their capacity to remove and retain AFM<sub>1</sub>.
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