Development of a simple FeSO₄-based liquid medium for reliable biofilm induction in Candida albicans.

Reproducibility in Candida albicans biofilm research is often hindered by variations in culture media and strain-dependent responses. This study evaluates the effect of oxygen-scavenging agents on biofilm induction to identify a simple and standardized liquid medium suitable for rapid and consistent biofilm formation. Modified Sabouraud Dextrose Broth (SDB) was supplemented with ferrous sulfate (FeSO₄), sodium sulfite (Na₂SO₃), ascorbic acid, and tested across four clinical isolates of C. albicans. FeSO₄-supplemented SDB promoted significant biofilm formation within 24 h and the highest biomass at 48 h (OD595 = 0.90-0.95), significantly greater than Na₂SO₃ (OD595 = 0.60-0.70) and ascorbic acid (OD595 = 0.30-0.45) (****p < 0.0001). The untreated control showed negligible biofilm formation. These findings establish FeSO₄-enriched SDB as a rapid, reproducible, and easily adaptable liquid medium for biofilm induction in C. albicans, providing a valuable tool for antifungal screening and biofilm-associated pathogenesis studies.