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In the present study, we examined antifungal activity of chitosan against Trichophyton rubrum (T. rubrum), which is a dermatophytic fungus and colonizes the upper layer of dead skin and causes athlete’s foot. Water-soluble chitosan oligosaccharide (AS-WSC), which had been prepared by the hydrolysis of high molecular weight water-soluble chitosan, was used in this study. The results upon screening, the AS-WSC inhibited T. rubrum, with zones of inhibition of 2% and 0.2% AS-WSC were 22 and 6 mm, respectively, while that of fluconazole was 30 mm. Minimal inhibitory concentration (MIC) values of AS-WSC was 1.25 mg/ml, while fluconazole showed MIC value of 2 μg/ml. Inhibition percentages of radial growth by 2% and 0.2% AS-WSC were 61% and 38%, respectively, while that of fluconazole was 73%. Two% and 0.2% AS-WSC treatment resulted in reduction of mycelium dry weights by 77% and 56%, respectively, while fluconazole reducing that by 84%. Our results suggest that AS-WSC can be a potential antifungal agent to treat infection by T. rubrum.