Voriconazole inhibits cross-kingdom interactions between Candida albicans and Actinomyces viscosus through the ergosterol pathway

Abstract

Candida albicans and Actinomyces viscosus are prominent microbes associated with dental root caries. The aim of this study was to investigate the effect of C. albicans on A. viscosus biofilms and to identify the mechanisms associated with this interaction. A. viscosus and C. albicans strains (wide-type and mutants) were used to form biofilms in vitro and in vivo, which were subsequently analysed by crystal violet assay and scanning electron microscopy (SEM) to investigate the effect of C. albicans on A. viscosus growth. A viable plate count and survival curve for C. albicans mutants and A. viscosus combinations were used to identify which C. albicans pathway was crucial for cross-kingdom interactions. Voriconazole was used to block their interactions both in vitro and in vivo. SEM, fluorescence in situ hybridisation (FISH), quantitative PCR and survival curve analyses were performed to evaluate the activity of voriconazole on C. albicans and A. viscosus interactions. The biomass and virulence of mixed-species biofilms were significantly enhanced compared with the A. viscosus biofilm alone. However, this was not observed in the mixed-species biofilms with the C. albicans mutant erg11Δ/Δ in vitro and in vivo, indicating that azoles may work on the mixed-species biofilms. As expected, voriconazole can effectively reduce the biomass of mixed-species biofilms. A high concentration of voriconazole (1 µg/mL) reduced the abundance of C. albicans, whilst a low voriconazole concentration (0.25 µg/mL) blocked their interactions similar to the effect of the erg11Δ/Δ mutant. Voriconazole may be a candidate strategy to combat root caries pathogens.