• Tea tree oil showed the strongest anti- Candida activity across all species and biofilm stages. • Whole essential oils consistently outperformed their isolated major components. • Biofilm susceptibility decreased markedly with increasing maturation (4-48 h). • Species-specific synergistic EO pairs were identified via checkerboard assays. • Optimized EO combinations inhibited 24 h Candida biofilms by over 90%. Candida biofilm-associated infections pose a major therapeutic challenge due to their high antifungal tolerance, species-specific pathogenicity, and rising resistance to conventional drugs. Essential oils (EOs) have emerged as promising natural antifungal agents, yet comparative studies evaluating whole oils and their major constituents across biofilms of different maturation stages remain limited. This study investigated the antibiofilm effects of tea tree, ceylon cinnamon bark, thyme, clove, and peppermint EOs, as well as their major components, against Candida albicans, C. glabrata, C. krusei , and C. tropicalis . Minimal inhibitory concentration was tested with microdilution assay. Biofilm inhibition was assessed at four maturation time points (4, 12, 24, and 48 h), and synergistic interactions were examined using checkerboard assays and Fractional Inhibitory Concentration Index (FICI) calculations. Tea tree oil was the most potent across all species and stages (e.g., 82.3% inhibition at 4 h vs. 40.0% at 48 h), followed by cinnamon bark and thyme oil. Whole EOs consistently outperformed their isolated components, indicating synergistic multi-component effects. Species-specific susceptibility patterns were apparent: C. krusei and C. glabrata were the most sensitive, whereas C. albicans exhibited the highest tolerance. Checkerboard assays identified strong synergism for tea tree-cinnamon oil against C. albicans and C. krusei (FICI 0.375) and for tea tree-thyme oil against C. tropicalis (FICI 0.49). These optimized EO combinations achieved exceptional inhibition of 24-hour biofilms (89-94%). Overall, EOs - particularly tea tree and its species-tailored combinations - represent promising candidates for disrupting Candida biofilms and may serve as effective components of future antifungal treatments.
Balázs et al. (Sun,) studied this question.