Dermatomycoses include superficial fungal infections of the skin and its appendages. Trichophyton rubrum, Candida albicans, and Candida parapsilosis are some of the most prevalent species that cause dermatomycoses. Several studies show a variable predominance of Candida spp. in relation to dermatophytes, especially in onychomycosis and the possibility of isolating both from the same site. The ability of dermatophytes to form biofilms recently been explored and there is currently no evidence on the involvement of these filamentous fungi in multi-species biofilms. Thus, this study aims to investigate the probable dual-species interaction between T. rubrum and C. albicans and T. rubrum and C. parapsilosis biofilms, considering variable formation conditions, as well as the susceptibility of these dual-species biofilms against terbinafine and efinaconazole. Three conditions of formation of dual-species biofilms were tested: (a) the suspensions of T. rubrum and Candida albicans or C. parapsilosis placed together; (b) suspensions of C. albicans and C. parapsilosis added the preadhesion of T. rubrum biofilms; (c) after the maturation of T. rubrum sessile cells. In the first and second conditions, the quantification of metabolic activities, biomass, and polysaccharide materials of mixed biofilms tended to resemble Candida monospecies biofilms. In the third condition, the profiles were modified after the addition of Candida, suggesting that T. rubrum biofilms served as substrate for the development of Candida biofilms. Scanning electron microscopy showed Candida predominance, however, numerous blastoconidia were noted, most evident in the conditions under which Candida was added after the pre-adhesion and maturation of T. rubrum biofilms. Despite the predominance of Candida, the presence of T. rubrum appears to inhibit C. albicans filamentation and C. parapsilosis development, confirming an antagonistic interaction. Fungal burden assays performed when the biofilms were formed together confirmed Candida predominance, as well as susceptibility to antifungals. Further studies will be needed to identify the components of the Candida and T. rubrum biofilm supernatants responsible for inhibiting dermatophyte growth and C. albicans filamentation.

Histoplasma capsulatum is a dimorphic fungus that causes an important systemic mycosis called histoplasmosis. It is an infectious disease with high prevalence and morbidity that affects the general population. Recently, the ability of these fungi to form biofilms, a phenotype that can induce resistance and enhance virulence, has been described. Despite some efforts, data regarding the impact of nutrients and culture media that affect the H. capsulatum biofilm development in vitro are not yet available. This work aimed to study H. capsulatum biofilms, by checking the influence of different culture media and oxygen atmospheres in the development of these communities. The biofilm formation by two strains (EH-315 and G186A) was characterized under different culture media: [Brain and Heart Infusion (BHI), Roswell Park Memorial Institute (RPMI) with 2% glucose, Dulbecco’s Modified Eagle’s Medium (DMEM) supplemented with 10% fetal bovine serum and nutrient medium HAM-F12 (HAM-F12) supplemented with glucose (18.2 g/L), glutamic acid (1 g/L), HEPES (6 g/L) and L-cysteine (8.4 mg/L)] and oxygen atmospheres (aerobiosis and microaerophilia), using the XTT reduction assay to quantify metabolic activities, crystal violet staining for biomass, safranin staining for the quantification of polysaccharide material and scanning electron microscopy (SEM) for the observation of topographies. Results indicated that although all culture mediums have stimulated the maturation of the communities, HAM-F12 provided the best development of biomass and polysaccharide material when compared to others. Regarding the oxygen atmospheres, both stimulated an excellent development of the communities, however in low oxygen conditions an exuberant amount of extracellular matrix was observed when compared to biofilms formed in aerobiosis, mainly in the HAM-F12 media. SEM images showed yeasts embedded by an extracellular matrix in several points, corroborating the colorimetric assays. However, biofilms formed in BHI, RPMI, and DMEM significantly induced yeast to hyphae reversal, requiring further investigation. The results obtained so far contribute to in vitro study of biofilms formed by these fungi and show that nutrition promoted by different media modifies the development of these communities. These data represent advances in the field of biofilms and contribute to future studies that can prove the role of these communities in the fungi-host interaction.

Histoplasma capsulatum is a fungus that causes histoplasmosis. The increased evolution of microbial resistance and the adverse effects of current antifungals help new drugs to emerge. In this work, fifty-four nitrofurans and indoles were tested against the H. capsulatum EH-315 strain. Compounds with a minimum inhibitory concentration (MIC90) equal to or lower than 7.81 µg/mL were selected to evaluate their MIC90 on ATCC G217-B strain and their minimum fungicide concentration (MFC) on both strains. The quantification of membrane ergosterol, cell wall integrity, the production of reactive oxygen species, and the induction of death by necrosis–apoptosis was performed to investigate the mechanism of action of compounds 7, 11, and 32. These compounds could reduce the extracted sterol and induce necrotic cell death, similarly to itraconazole. Moreover, 7 and 11 damaged the cell wall, causing flaws in the contour (11), or changing the size and shape of the fungal cell wall (7). Furthermore, 7 and 32 induced reactive oxygen species (ROS) formation higher than 11 and control. Finally, the cytotoxicity was measured in two models of cell culture, i.e., monolayers (cells are flat) and a three-dimensional (3D) model, where they present a spheroidal conformation. Cytotoxicity assays in the 3D model showed a lower toxicity in the compounds than those performed on cell monolayers. Overall, these results suggest that derivatives of nitrofurans and indoles are promising compounds for the treatment of histoplasmosis.