The biofilms of filamentous-forming fungi are a novel and still insufficiently understood research topic. We have studied Aspergillus fumigatus, an ubiquitous opportunistic pathogenic fungus, as a representative model for a study of biofilm formation by filamentous fungi and for assessing the potential anti-biofilm activity of natural substances. The activity of antibiotic amphotericin B and selected natural substances: baicalein, chitosan and rhamnolipid was studied. The minimum suspension inhibitory concentrations (MIC) were determined and the biofilm susceptibility was investigated by determining the metabolic activity of sessile cells (XTT assay) and total biofilm biomass (crystal violet staining). Significant time-dependent differences in substances' anti-biofilm activity were observed. Images of A. fumigatus biofilm were obtained by Cellavista automatic light microscope and spinning disc confocal microscopy. Baicalein and rhamnolipid were not found as suitable substances for inhibition of the A. fumigatus biofilm formation, as neither of the substances inhibited the sessile cells metabolic activity or the total biofilm biomass even at tenfold MIC after 48 h. In contrast, chitosan at 10 × MIC (25 µg mL), suppressed the biofilm metabolic activity by 90 % and the total biofilm biomass by 80 % even after 72 h of cultivation. Amphotericin B inhibited only 14 % of total biofilm biomass (crystal violet staining) and 35 % of metabolic activity (XTT assay) of adherent cells under the same conditions. Our results therefore suggest chitosan as potential alternative for treating A. fumigatus biofilm-associated infections.

This study focuses on the isolation of a consortium of microorganisms from spontaneously fermenting must that naturally contain lactic acid bacteria, non-saccharomyces yeasts, and saccharomyces yeasts. To collect the greatest diversity of microorganisms, the consortium was taken from the point of micro-sparkling. Based on the growth curves, isolation was performed using individual special nutrient media, and the isolates were subsequently multiplied in the nutrient medium. Individual isolates were then used for fermentation tests to monitor the percentage of fermented sugar and hydrogen sulphide production. The highest fermentation abilities were achieved in the isolates containing Saccharomyces cerevisiae and Zygosaccharomyces bailii. The smallest amount of ethanol was formed from the isolates containing Hanseniaspora uvarum, while Candida sake isolate produced the lowest amount of hydrogen sulphide and Zygosaccharomyces bailii produced the highest. The other isolates produced an average amount. Based on these results, a consortium containing the given isolates in a certain ratio was compiled.