Abstract
The isolated protein-polysaccharide fraction (AAF) from the coelomic fluid of Dendrobaena veneta earthworm shows effective activity against Candida albicans yeast. Fungal cells of the clinical strain after incubation with the active fraction were characterized by disturbed cell division and different morphological forms due to the inability to separate the cells from each other. Staining of the cells with acridine orange revealed a change in the pH of the AAF-treated cells. It was observed that, after the AAF treatment, the mitochondrial DNA migrated towards the nuclear DNA, whereupon both merged into a single nuclear structure, which preceded the apoptotic process. Cells with a large nucleus were imaged with the scanning electron cryomicroscopy (Cryo-SEM) technique, while enlarged mitochondria and the degeneration of cell structures were shown by transmission electron microscopy (TEM). The loss of the correct cell shape and cell wall integrity was visualized by both the TEM and SEM techniques. Mass spectrometry and relative quantitative SWATH MS analysis were used to determine the reaction of the C. albicans proteome to the components of the AAF fraction. AAF was observed to influence the expression of mitochondrial and oxidative stress proteins. The oxidative stress in C. albicans cells caused by the action of AAF was demonstrated by fluorescence microscopy, proteomic methods, and XPS spectroscopy. The secondary structure of AAF proteins was characterized by Raman spectroscopy. Analysis of the elemental composition of AAF confirmed the homogeneity of the preparation. The observed action of AAF, which targets not only the cell wall but also the mitochondria, makes the preparation a potential antifungal drug killing the cells of the C. albicans pathogen through apoptosis.
Highlights
The isolated protein-polysaccharide fraction (AAF) from the coelomic fluid of Dendrobaena veneta earthworm shows effective activity against Candida albicans yeast
Control and AAF-treated C. albicans cells were stained with acridine orange
Previous studies have shown that the fraction of D. veneta coelomic fluid is active against various strains of C. albicans, i.e., a clinical strain and ATCC 10231, C. krusei, and ATCC 6 25830
Summary
The isolated protein-polysaccharide fraction (AAF) from the coelomic fluid of Dendrobaena veneta earthworm shows effective activity against Candida albicans yeast. The phenomenon is observed especially in intensive care units in Europe and the U SA8–11, where Candida albi‐ cans is most frequently responsible for fungal infections. It can induce superficial or systemic candidiasis. C. albicans can cause life-threatening infections due to its ability to alter its growth model In normal conditions, this microorganism grows as a commensal on the surface of the mucous membranes of the mouth, vagina, and skin and as part of the intestinal m icroflora[14,18]. The antibiotics used in candidiasis treatment have many side effects, which are especially dangerous to patients with a weakened immune system by e.g., chemotherapy or HIV infection. There are an increasing number of Candida strains that are resistant to conventionally used antibiotics, e.g., azoles or e chinocandins[10,14]
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