Background: Increase in resistance of Candida species, to routinely used antifungal agents has necessitated the quest for new drugs. Few studies have revealed that cow's urine can suppress the growth of pathogenic fungi. However there is no published report on antifungal effects of cow's urine on clinical Candida isolates. Objective: The present study aims at exploring the antifungal potential of cow's urine on clinical isolates of Candida species. Materials and methods: In this in-vitro experimental study four standard strains and 37 clinical isolates of Candida species were tested for their susceptibility to amphotericin B, fluconazole and voriconazole, by disk diffusion method. Detection of MIC of cow's urine for the Candida isolates was done by agar dilution method using 20–50% concentration of cow's urine. Results: Clinical isolates of Candida albicans n = 22 (59.5%) Candida glabrata n = 6 (16.2%), Candida tropicalis n = 3 (8.1%) and other Candida species n = 6 were tested for their antifungal susceptibility. Among them, 18.9% were resistant to voriconazole, 24.3% to amphotericin B and 35.1% to fluconazole. Statistically significant association was observed between susceptibility of voriconazole and that of cow's urine (p = 0.045). C. albicans ATCC14053, Candida parapsilosis ATCC22019 and 75.7% of clinical isolates of Candida were susceptible to cow's urine. Conclusion: Cow's urine distillate has concentration dependent inhibitory effect on Candida species and is effective on the isolates that are either resistant or sensitive to the routinely used antifungal agents.