Objective: Indonesia with high humidity and rainfall strongly supports the growth of fungi. The incidence of fungal infections has increased dramatically over the past three decades. Several factors including microbiological resistance led to the failure of antifungal therapy. This research was to evaluate the antifungal activity of extract and fraction of Auricularia auricula against the test fungus.Methods: Antifungal activity was performed by microdilution method using extract and n-hexane fraction, ethyl acetate fraction, methanol, and water fraction of A. auricula. MIC was determined by observing the turbidity compared with the control solution. Minimum fungicidal concentration (MFC) was determined by the presence of fungi colonies growth on medium after incubation for 24 h.Results: The fractions of A. auricular have antifungal activity against Candida albicans, with MIC 512 μg/ml and MFC 512 μg/ml; Microsporum gypseum with MIC 256 μg/ml in the extract; MIC 512 μg/ml and MFC 512 μg/ml of ethyl acetate fraction; MIC 256 μg/ml and MFC 512 μg/ml in n-hexane fraction; and Aspergillus flavus with MIC 2048 μg/ml and MFC 2048 μg/ml in the extract and ethyl acetate fraction. The most potent activity on C. albicans was exhibited by ethyl acetate fraction with MIC and MFC 512 μg/ml. Scanning electron microscopy showed changes in the structure of fungi cells after contact with the test solution.Conclusion: The best antifungal activity of A. auricula was ethyl acetate fraction against C. albicans
