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Although fumonisin B1 (FB1) is a mycotoxin poorly studied in wheat, the Fusarium genus fungi have been frequently detected in this cereal, especially in subtropical regions such as southern Brazil, thus studies on their occurrence are required. This study evaluated the mycobiota, water activity and the presence of FBB1 in 11 wheat cultivars grown in the northeastern and northwestern of Rio Grande do Sul. The mycobiota was identified by morphological and molecular techniques and FBB1 was determined by liquid chromatography with fluorescence detection (HPLC-FL). Microbial counts lower than 103CFU were found in the samples, and the fungi genera Phoma, Cladosporium and Fusarium were the most frequent. Despite this, 54% of the samples were contaminated with FB1 at levels ranging from 958 to 4,906μg FBB1 kgB-1 [ABSTRACT FROM AUTHOR]