Otomycosis is a fungal infection of the ear predominantly caused by Candida and Aspergillus spp. The possible virulence factors of Candida spp. are enzymes, such as proteases, phospholipases, phosphatases and esterase. According to our knowledge, protease production in Candida strains isolated from patients with otomycosis has not been investigated. The present study was aimed at determining in vitro protease activity in 8 strains of Candida spp. (C. parapsilosis, C. famata, C. guilliermondii and C. albicans) isolated from children with otomycosis. A majority of isolated strains 7/8 (87.5%) were protease positive. The protease activity ranged from Pz 0.61 to 0.78. Further investigation is necessary to clarify the contribution of protease production to Candida virulence associated with otomycosis.