High-throughput assay for effect screening of amphotericin B and bioactive components on filamentous Candida albicans

Aims The aim of this study was to develop a high-throughput robotic microtiter plate-based screening assay for Candida albicans, optimizing growth conditions to replicate the filamentous biofilm growth found in vivo, and subsequently, to demonstrate the assay by evaluating the effect of nutritional drinks alone and in combination with the antifungal amphotericin B (AmB). Methods and Results Candida albicans cultured in a defined growth medium showed filamentous growth in microcolonies, mimicking the morphology of oral mucosal disease (oral candidiasis). Addition of nutrient drinks containing fruit juices, fish oil and whey protein to the medium resulted in changed morphology and promoted growth as free yeast cells and with weak biofilm structures. Minimum inhibitory concentration of AmB on the biofilms was 0.25 mu g ml(-1), and this was eightfold reduced (0.0038 mu g ml(-1)) in the presence of the nutritional drinks. Conclusions The established assay demonstrated applicability for screening of antifungal and anti-biofilm effects of bioactive substances on C. albicans biofilm with clinically relevant morphology. Significance and Impact of the Study Candida albicans is the causative agent of the majority of fungal infections globally. The filamentous morphology of C. albicans and the ability to form biofilm are traits known to increase virulence and resistance towards antifungals. This study describes the development of a plate-based in vitro screening method mimicking the filamentous morphology of C. albicans found in vivo. The assay established can thus facilitate efficient antifungal drug discovery and development.

Automated summary

This study developed a high-throughput robotic microtiter plate-based screening assay that mimics the filamentous morphology of C. albicans. The assay demonstrated the effectiveness of nutritional drinks in promoting the growth of C. albicans and showed that amphotericin B had a reduced inhibitory effect in the presence of these drinks. This assay can facilitate efficient antifungal drug discovery and development.