The study evaluated the change in the clinical antifungal sensitivity profile of A. flavus strains after exposure to azole and benzimidazole fungicide. Exposure to fungicide altered the sensitivity profile for the antifungal itraconazole, voriconazole and posaconazole. This change was characterized by an increase in the minimum inhibitory concentration (MIC) from 16 to 32 times, evidencing the development of resistance phenotypes. The most significant changes were found after exposure to a pool of the fungicide with MIC of up to 256 times, which is considered, to the best of our knowledge, the first case report of such a high level of resistance induced by azole fungicide exposure. This observation probably indicates a synergistic action among azole compounds that potentiates the development of resistance phenotypes. In addition, exposure to fungicide changed the pigmentation of the colonies from green to white. The development of resistance to fungicides represents risks to human health, since azole fungicides are used widely in the agriculture, and a single agricultural fungicide spray often includes more than one azole compound.