METHODS: The antifungal susceptibility of hydroxychavicol (HC) and Amphotericin B (AMB) separately and in a mixture (HC + AMB) against Candida albicans ATCC 14053, Candida parapsilosis ATCC 22019, Candida tropicalis ATCC 13803, and Candida dubliniensis ATCC MYA-2975 was determined by broth microdilution technique. Minimal Inhibitory Concentration was calculated based on the CLSI protocols. The MIC50, fractional inhibitory concentration (FIC) index, and IC50 were also determined. The IC50 values were used as the treatment concentration of HC, AMB, and HC + AMB to study the effect of antifungal inhibition on yeast hypha transition (gemination). The germ tube formation percentage of candida species was calculated at several intervals using a colorimetric assay.
RESULTS: The MIC50 range of HC alone against Candida species was between 120-240 µg per mL while that of AMB was between 2-8 µg per mL, respectively. The combination of HC + AMB at 1:1 and 2:1 demonstrated the strongest synergistic activity against C. albicans with an FIC index of 0.07. Moreover, within the first hour of treatment, the total percentage of germinating cells was significantly reduced by 79% (p < 0.05).
CONCLUSION: The combination of HC + AMB displayed synergism and inhibited C. albicans hyphal growth. HC + AMB combination slowed the germination process and exhibited consistent prolonged effect up to 3 h post-treatment. The results of this study will pave the way for potential in vivo studies.