Wheat, barley and maize are the mainly consumed cereals in Tunisia. This study aimed to determine the mycoflora of these cereals with special focus on the mycotoxigenic and species. Freshly harvested samples and other stored samples of each type of cereal (31 and 34 samples, respectively) were collected in Tunisia and cultured for fungal isolation and identification. Identification of fungal genera was based on morphological and species were identified by species specific PCR assays complemented with DNA sequencing. (70.83%), (62.50%), (54.17%) and (41.67%) were the most frequent fungi isolated from wheat. (75%), (70%), (65%) and (65%) were the most frequently recovered genera from barley. The predominant genera in maize were (76.19%), (42.86%), and (38.09%)., , and were detected in both stored and freshly harvested grain samples. The frequencies of contamination with, and were higher in freshly harvested samples, whereas species were more frequent in stored samples. The predominant species detected were and. The species detected were, ,, and . This study suggested the potential risk for Aflatoxins and, to a lesser extent, for Ochratoxin A in Tunisian cereals. This is the first survey about mycoflora associated with wheat, barley and maize in Tunisia.

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