Candida tropicalis biofilm formation and expression levels of the CTRG ALS-like genes in sessile cells.

Candida tropicalis is an emergent pathogen with a high rate of mortality associated with it; however, less is known about its pathogenic capacity. Biofilm formation (BF) has important clinical repercussions, and it begins with adherence to a substrate. The adherence capacity depends principally on the cell surface hydrophobicity (CSH) and, at a later stage, on specific adherence due to adhesins. The ALS family in C. tropicalis, implicated in adhesion and BF, is represented in several CTRG genes. In this study, we determined the biofilm-forming ability, the primary adherence, and the CSH of C. tropicalis, including six isolates from blood and seven from urine cultures. We also compared the expression of four CTRG ALS-like genes (CTRG_01028, CTRG_02293, CTRG_03786, and CTRG_03797) in sessile versus planktonic cells, selected for their possible contribution to BF. All the C. tropicalis strains were biofilm producers, related to its filamentation capacity; all the strains displayed a high adherence ability correlated to the CSH, and all the strains expressed the CTRG genes in both types of growth. Urine isolates present, although not significantly, higher CSH, adherence, and biofilm formation than blood isolates. This study reveals that three CTRG ALS-like genes-except CTRG_03797-were more upregulated in biofilm cells, although with a considerable variation in expression across the strains studied and between the CTRG genes. C. tropicalis present a high biofilm capacity, and the overexpression of several CTRG ALS-like genes in the sessile cells suggests a role by the course of the biofilm formation.

Antifungal and anti-biofilm activity of a new Spanish extract of propolis against Candida glabrata.

BACKGROUND: Resistance to traditional antifungal agents is a considerable health problem nowadays, aggravated by infectious processes related to biofilm formation, usually on implantable devices. Therefore, it is necessary to identify new antimicrobial molecules, such as natural products, to develop new therapeutic strategies to prevent and eradicate these infections. One promising product is propolis, a natural resin produced by honeybees with substances from various botanical sources, beeswax and salivary enzymes. The aim of this work was to study the effect of a new Spanish ethanolic extract of propolis (SEEP) on growth, cell surface hydrophobicity, adherence and biofilm formation of Candida glabrata, a yeast capable of achieving high levels of resistance to available anti-fungal agents. METHODS: The antifungal activity of SEEP was evaluated in the planktonic cells of 12 clinical isolates of C. glabrata. The minimum inhibitory concentration (MIC) of propolis was determined by quantifying visible growth inhibition by serial plate dilutions. The minimum fungicide concentration (MFC) was evaluated as the lowest concentration of propolis that produced a 95% decrease in cfu/mL, and is presented as MFC50 and MFC90, which corresponds to the minimum concentrations at which 50 and 90% of the C. glabrata isolates were inhibited, respectively. Influence on cell surface hydrophobicity (CSH) was determined by the method of microbial adhesion to hydrocarbons (MATH). The propolis effect on adhesion and biofilm formation was determined in microtiter plates by measurement of optical density (OD) and metabolic activity (XTT-assay) in the presence of sub-MIC concentrations of SEEP. RESULTS: SEEP had antifungal capacity against C. glabrata isolates, with a MIC50 of 0.2% (v/v) and an MFC50 of 0.4%, even in azole-resistant strains. SEEP did not have a clear effect on surface hydrophobicity and adhesion, but an inhibitory effect on biofilm formation was observed at subinhibitory concentrations (0.1 and 0.05%) with a significant decrease in biofilm metabolism. CONCLUSIONS: The novel Spanish ethanolic extract of propolis shows antifungal activity against C. glabrata, and decreases biofilm formation. These results suggest its possible use in the control of fungal infections associated with biofilms.

[Cell surface hydrophobicity as an indicator of other virulence factors in Candida albicans]. / La hidrofobicidad de la superficie celular como indicador de otros factores de virulencia en Candida albicans.

OBJECTIVE: To evaluate virulence factors involved in the adhesion process, such as cell surface hydrophobicity (CSH), adherence to plastic capacity, adherence capacity to buccal epithelial cells (BEC), and biofilm formation, in 17 strains of C. albicans isolated from bronchial aspirates of critically ill patients. METHOD: The CSH of the strains of C. albicans was determined using the MATH method, a microbial adhesion to hydrocarbons test. The study of adherence to plastic was performed in microtitre plates in accordance with Christensen's technique. Biofilm formation was studied in polystyrene microtitre plates, according to the method of Ramage. Adherence to BEC was evaluated by quantifying the percentage of adhered yeasts to cells. RESULTS: All the strains studied showed factors directly involved in adhesion, with variability in the degree of expression among them. Medium-high levels of CSH were found in 52.9% of the strains. The percentage of strains with high values in adherence to plastic was 35.3%. The most hydrophobic strains were the most adherent to plastic, with a correlation coefficient of 0.76. Of the 12 biofilm-producing strains, 6 were high producers. These strains had also high levels of CSH and adherence to plastic, with significant results. All the strains studied adhered to BEC, with results ranging widely from 45 to 157 yeasts/100 BEC, with no significant correlation with the rest of the parameters studied, although CSH was seen to be an indispensable prior requisite for adherence to cells. CONCLUSION: CSH is a variable characteristic in C. albicans and is directly related to adherence to plastic and biofilm formation. Ease in evaluating CSH permits its quantification, and could be used as an indicator of the presence of other determinants of pathogenicity.