Atypical changes in Candida albicans cells treated with the Venetin-1 complex from earthworm coelomic fluid

Abstract

In the present research, the effect of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworm on Candida albicans cells was characterized. The compound destroyed fungal cells without showing cytotoxicity to human skin fibroblasts, which was demonstrated in earlier studies. Since it had an effect on the fungal cell wall and membrane, this complex was compared with the known antifungal antibiotic fluconazole. Both preparations disturbed the division of yeast cells and resulted in the formation of aggregates and chains of unseparated cells, which was illustrated by staining with fluorochromes. Fluorescent staining of the cell wall with Calcofluor white facilitated comparison of the types of aggregates formed after the action of both substances. The analysis performed with the use of Congo red showed that Venetin-1 exposed deeper layers of the cell wall, whereas no such effect was visible after the use of fluconazole. The FTIR analysis confirmed changes in the mannoprotein layer of the cell wall after the application of the Venetin-1 complex. Staining with Rhodamine 123 and the use of flow cytometry allowed comparison of changes in the mitochondria. Significantly elongated mitochondria were observed after the Venetin-1 application, but not after the application of the classic antibiotic. Phase contrast microscopy revealed vacuole enlargement after the Venetin-1 application. The flow cytometry analysis of C. albicans cells treated with Venetin-1 and fluconazole showed that both substances caused a significant decrease in cell viability.