Studies have shown evidence of human exposure to aflatoxin M₁ due to the consumption of contaminated milk and dairy products (mainly cheeses). This poses a great risk to public health, since milk and milk products are frequently consumed by a portion of the population considered immunosuppressed, children and the elderly. Knowledge of the negative impacts of aflatoxins on health and economics has led to investigations of strategies to prevent their formation in food, as well as to eliminate, inactivate or reduce the bioavailability of these toxins in contaminated products This study evaluated the effect of microbiological methods using lactic acid bacteria on aflatoxin M₁ (AFM₁) reduction in Minas Frescal cheese (typical Brazilian product, being among the most consumed cheeses in Brazil) spiked with 1 µg/L AFM₁. Inactivated by heat treatment, lactic acid bacteria (0.5 vol/vol % of L. rhamnosus and L. lactis) were added during the cheese production process. Nine cheeses were produced, divided into 3 treatments: negative controls (without AFM₁ or lactic acid bacteria), positive controls (AFM₁ only), and lactic acid bacteria + AFM₁. Samples of cheese were collected on d 2, 10, 20 and 30 after the date of production and submitted to composition analyses and determination of AFM₁ by high performance liquid chromatography. The directional reductions of AFM₁ in cheese by lactic acid bacteria indicate a potential application of inactivated lactic acid bacteria in reducing the bioavailability of AFM₁ in Minas frescal cheese without physical-chemical and microbiological modifications during the 30-d experimental period.