Introduction: Onychomycosis presents significant therapeutic challenges, with high recurrence rates and the need for prolonged treatment adherence. While previous studies have explored the contamination risks of cosmetic nail polishes, little is known about the microbiological safety of antifungal nail polishes used in onychomycosis treatment. Methods: This study investigated potential contamination in antifungal nail polish bottles containing ciclopirox olamine 8% (CO) and amorolfine 5% (AM) used by patients diagnosed with onychomycosis confirmed by mycological exams. After treatment, applicators were analyzed using Sabouraud and Mycosel® culture media. Results: Of the 67 randomly selected nails treated, 100% showed positive direct mycological examination, and 49% had fungal growth in culture. However, none of the 67 applicators (49 CO and 18 AM) exhibited fungal growth. Conclusion: The study findings suggest that antifungal nail polishes do not act as contamination reservoirs, likely due to their inherent antifungal properties. Nonetheless, patients should be educated on proper hygiene to minimize cross-contamination risks.

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