We developed a niosomal formulation of diallyl sulfide (DAS), a garlic oil component, and evaluated its efficacy against experimental candidiasis in mice. DAS-bearing niosomes prepared from sorbitan monoester surfactants were evaluated for drug entrapment efficiency, release kinetics, toxicity, size, ζ-potential and others. Mice challenged with Candida albicans were treated with various DAS formulations. The efficacy of the formulations was assessed on the basis of reduction in mortality and decrease in residual fungal load in vital organs, such as liver and spleen, of treated mice. Niosomal DAS (12 mg/kg body weight) significantly reduced fungal load and mortality in treated animals compared with the free form of DAS. Niosomal DAS was also found to be free of toxic manifestations, as revealed by histopathological studies, as well as liver/kidney function tests. Incorporation of DAS in niosomes enhances its antifungal efficacy. Further studies are needed to optimize the current findings to develop an efficient nature-derived alternative antifungal therapeutic strategy.