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ntroduction: Otomycosis which is caused by saprophytic fungi can be treated by topical antifungal agents. Sometimes eradication of the disease is difficult in patients with risk factors. Considering controversies about the anti-microbial effect of cerumen, this research was performed in Babul, was focused on revealing the antifungal effect of cerumen of healthy people on fungi causing otomycosis. Materials and Methods: This experimental study was carried out on the 60 samples of healthy peopleُ s ear canal secretion. The average ages of the subjects were between 2-85 years. The antifungal activity of cerumen solution on 4 species of fungi including; Aspergillus fumigatus, Aspergillus niger, Candida albicans (clinical isolate) and standard Candida albicans were analyzed by paper disc diffusion and micro dilution methods. Results: 10% of cerumen showed antifungal activity in paper disc diffusion; whereas in the micro dilution method all of samples had antifungal activity. The most antifungal effect observed was on Aspergillus niger (27 samples) and the least effect on the species of Candida albicans (16 samples). Conclusion: According to the obtained results, cerumen has different effects on fungal growth, thus it is necessary to study more about components of cerumen for new approaches in future.