This study evaluated the antifungal efficacy of gentian violet (GV) in an experimental vulvovaginal candidiasis (VVC) model. susceptibility and cytotoxicity assays were performed to validate the antifungal potential and safety of GV. The antifungal efficacy was then evaluated through comparative analysis of the fungal burden following treatment with GV or nystatin, as well as assessment of the vaginal tissue by histology and electron microscopy. GV demonstrated a safe antifungal profile against , with a significant decrease in fungal burden and an improvement in the inflammatory process evaluated histologically. The results of this study motivate further assessment of GV as a promising alternative for VVC therapy.