Annual and Seasonal Variations in Aflatoxin M1 in Milk: Updated Health Risk Assessment in Serbia

Aflatoxin M1 (AFM1), a hepatocarcinogenic metabolite of aflatoxin B1, poses significant risks to human health through its presence in milk and dairy products. This study investigates AFM1 contamination in raw milk produced in Serbia from 2021 to 2025, assessing annual and seasonal variations and associated health risks. A total of 907 milk samples were analyzed using enzyme-linked immunosorbent assay (ELISA), revealing contamination in 70.1% of samples, with mean concentrations exceeding the EU regulatory limit of 50 ng/kg. Seasonal analysis identified the highest contamination levels during winter, attributed to increased use of contaminated feed during colder months. Health risk assessments estimated the daily intake of AFM1 and associated health risks, with high-exposure individuals showing notably reduced margins of safety. The research demonstrates the essential requirement for better feed quality management alongside enhanced regulatory oversight along with health programs that reduce AFM1 exposure in Serbian populations.