Viola odorata L. is an important medicinal plant with antioxidant, anti-inflammatory, antibacterial, antifungal and anticancerous activities. The present study was carried out to evaluate the antimicrobial potential of acetone, ethanol, methanol and ethyl acetate extracts of V. odorata against five bacterial strains (Bacillus subtilis, Micrococcus luteus, Bacillus cereus, Klebsiella pneumonia and Escherichia coli) by the agar diffusion method and two fungal strains (Bipolaris specifera and Fusarium solani) by poisoned food technique. The results revealed that the ethanol extract was the most effective against the tested pathogens and thus V. odorata can be used as a potential source for the development of drugs.