Intra-examiner reproducibility of the agar plate method for estmation of phospholipase activity in candida albicans

Although measurement errors can impair statistical analysis, reliability analysis has been neglected in applied microbiology. This study assessed the intra-rater reproducibility of the Agar-based method for estimation of phospholipase activity (Pz). Pz readings were performed twice by two examiners (E1, E2), either directly on plates or in photos, and both black and white backgrounds were used. Pz values were taken from one or triplicate colonies from each sample (n=30). Intra-examiner reproducibility was estimated using Intraclass Correlation Coefficient (ICC). For both examiners, reading triplicate (ICCE1=0.91, ICCE2=0.86) was better than one colony (ICCE1=0.86, ICCE2=0.80). E1 had an excellent concordance when measurements were performed on photos using a white background (ICC=0.95) and a good concordance in the other conditions

Cell membrane integrity of candida albicans after different protocols of microwave irradiation

To evaluate the ability of low time microwaveexposureto inactivate and damage cell membrane integrity of C. albicans. Materials and Methods: Two 200ml C. albicans suspensions were obtained. Sterile dentures were placed in a beaker containing Experimental (ES) or Control suspensions (CS). ES was microwaved at 650 W for 1, 2, 3, 4 or 5 min. Suspensions were optically counted using Methylene blue dye as indicative of membrane-damaged cells; spread on Agar Sabouraud dextrose (ASD) for viability assay; or spectrophotometrically measured at 550nm. Cell-free solutions were submitted to content analyses of protein (Bradford and Pyrogallol red methods); Ca++ (Cresolphthalein Complexone method); DNA (spectrophotometer measurements at 260nm) and K+ (selective electrode technique). Data were analyzed by Student-t test and linear regression (α=0.05). In addition, flowcytometry analysis of Candida cells in suspensionwas performed using propidium iodide. Results: All ES cells demonstrated cell membrane damage at 3, 4 and 5 min,viable cells were nonexistent at 3, 4 and 5 min ES ASD plates and optical density of ES and CS was not significantly differentfor all exposition times. ES cells released highcontents of protein, K+ , Ca++ and DNA after 2 min exposition when compared to that of the CSs. Similar results were observed with flow cytometry analysiswith regard to the periodsof microwave exposure. Conclusions: Microwave irradiation inactivated C. albicansafter 3min and damaged cell membrane integrity after 2 min exposition.

Prevalence of Candida spp. during radiographic examination in Diabetes mellitus patients

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Biofilmes multiespécies de Candida Albicans associados com streptococcus mitis e streptococcus sanguinis: estudo in vitro e in vivo

Pós-graduação em Odontologia Restauradora - ICT

Susceptibility of Candida albicans and Candida glabrata biofilms to silver nanoparticles in intermediate and mature development phases

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Dynamics of biofilm formation and the interaction between candida albicans and methicillin-susceptible (MSSA) and -resistant staphylococcus aureus (MRSA)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

In vivo evaluation of photodynamic inactivation using Photodithazine (R) against Candida albicans

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Experimental Autoimmune Encephalomyelitis Development Is Aggravated by Candida albicans Infection

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Evaluation of gene expression SAP5, LIP9, and PLB2 of candida albicans biofilms after photodynamic inactivation

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Inactivation of candida albicans by cold atmospheric pressure plasma jet

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Nanostructured lipid system as a strategy to improve the anti-Candida albicans activity of Astronium sp

The genus Astronium (Anacardiaceae) includes species, such as Astronium fraxinifolium, Astronium graveolens, and Astronium urundeuva, which possess anti-inflammatory, anti-ulcerogenic, healing, and antimicrobial properties. Nanostructured lipid systems are able to potentiate the action of plant extracts, reducing the required dose and side effects and improving antimicrobial activity. This work aims to evaluate a nanostructured lipid system that was developed as a strategy to improve the anti-Candida albicans activity of hydroethanolic extracts of stems and leaves from Astronium sp. The antifungal activity against C. albicans (ATCC 18804) was evaluated in vitro by a microdilution technique. In addition to the in vitro assays, the Astronium sp. that showed the best antifungal activity and selectivity index was submitted to an in vivo assay using a model of vulvovaginal candidiasis infection. In these assays, the extracts were either used alone or were incorporated into the nanostructured lipid system (comprising 10% oil phase, 10% surfactant, and 80% aqueous phase). The results indicated a minimal inhibitory concentration of 125.00 µg/mL before incorporation into the nanostructured system; this activity was even more enhanced when this extract presented a minimal inhibitory concentration of 15.62 µg/mL after its incorporation. In vivo assay dates showed that the nanostructure-incorporated extract of A. urundeuva leaves was more effective than both the unincorporated extract and the antifungal positive control (amphotericin B). These results suggest that this nanostructured lipid system can be used in a strategy to improve the in vitro and in vivo anti-C. albicans activity of hydroethanolic extracts of Astronium sp.

Effect of tyrosol on adhesion of Candida albicans and Candida glabrata to acrylic surfaces

The prevention of adhesion of Candida cells to acrylic surfaces can be regarded as an alternative to prevent denture stomatitis. The use of quorum sensing molecules, such as tyrosol, could potentially interfere with the adhesion process. Therefore, the aim of this study was to assess the effect of tyrosol on adhesion of single and mixed cultures of Candida albicans and Candida glabrata to acrylic resin surfaces. Tyrosol was diluted in each yeast inoculum (10(7) cells/ml in artificial saliva) at 25, 50, 100, and 200 mM. Then, each dilution was added to wells of 24-well plates containing the acrylic specimens, and the plates were incubated at 37°C for 2 h. After, the effect of tyrosol was determined by total biomass quantification, metabolic activity of the cells and colony-forming unit counting. Chlorhexidine gluconate (CHG) was used as a positive control. Data were analyzed using analysis of variance (ANOVA) and the Holm-Sidak post hoc test (α = 0.05). The results of total biomass quantification and metabolic activity revealed that the tyrosol promoted significant reductions (ranging from 22.32 to 86.16%) on single C. albicans and mixed cultures. Moreover, tyrosol at 200 mM and CHG significantly reduced (p < 0.05) the number of adhered cells to the acrylic surface for single and mixed cultures of both species, with reductions ranging from 1.74 to 3.64-log10. In conclusion, tyrosol has an inhibitory effect on Candida adhesion to acrylic resin, and further investigations are warranted to clarify its potential against Candida infections.

Low-temperature plasma: an effective approach against Candida albicans biofilm

This study evaluated the antifungal potential of low-temperature plasma (LTP) on a 72-hour Candida albicans biofilm. A growth inhibition zone test was conducted with agar plates inoculated with C. albicans and submitted to LTP and argon application at 3 and 10 mm for 10, 30, 60, 90, and 120 seconds. The groups for biofilm assays were 60 seconds of LTP application with a tip-to-sample distance of 3 mm (LTP-3) and 10 mm (LTP-10); –application of only argon gas for 60 seconds with a tip-to-sample distance of 3 mm (Ar-3) and 10 mm (Ar-10); and no treatment. The C. albicans biofilm was grown on saliva-coated discs. The medium was replaced every 24 hours. Confocal laser scanning microscopy revealed the proportion of live and dead cells, and variable pressure scanning electron microscopy (VPSEM) showed biofilm/cell structure. No inhibition zone was observed for control and either Ar groups. For the LTP groups, a progressively increasing of inhibition zone diameter was observed for different treatment durations. The LTP-3 and LTP-10 groups presented higher proportions of dead cells compared with the Ar-3 and Ar-10 groups. VPSEM revealed cell perforations in the LTP-3 and LTP-10 groups. A short period of LTP exposure demonstrated an antifungal effect on C. albicans biofilm.

Genetic evaluation and activity of antifungal against clinical isolate Candida albicans biofilms

Candida yeasts are common in the oral cavity and can cause candidosis in the presence of predisposing factors, especially diabetes. The manifestation of the disease is related to this set of local factors such as the presence of dental prostheses, salivary pH, salivary flow and tobacco and the ability to form biofilms. Biofilms are specific and organized communities of cells under the control of signaling molecules rather than random accumulations of cells resulting from cell division and frequently are drugs resistance. Aim: The objectives of this study were to determine the genetic patterns of these C. albicans isolates and to evaluate the in vitro activity amphotericin B and caspofungin against C. albicans biofilms. Methods: Microbial samples were collected from subgingival sites and seeded in CHROMagar for subsequent identification of C. albicans by PCR. Genotypes were defined based on the identification of the transposable introns in the 25S rDNA by PCR. Results: In this study, 6 strains were identified as C. albicans and of these, 3 strains were genotype A and 3 were genotype B. The results showed that both amphotericin B and caspofungin exhibited strong antifungal activities against C. albicans biofilm formation and inhibiting the biofilm formation ranging from 70.8 – 95.3% and 77.7 - 88.7%, respectively. The antifungals studied had low inhibitory effect on preformed biofims, ranging from 39.5 - 50.8% for amphotericin B and from 23.1 - 36.9% for caspofungin at the same concentration. The activity of the two drugs was most effective in inhibit biofilm formation.

In vitro antifungal susceptibility of candida albicans isolates from patients with chronic periodontitis and diabetes

Alterations that lead to deficiency of the immune system, such as diabetes mellitus, may promote proliferation of Candida albicans and selection of strains which have greater ability to adhere and to penetrate the host tissues. Recent studies indicate an increase of the antifungal resistance of C. albicans isolates in periodontal pockets, suggesting that the oral cavity could be a reservoir of resistant yeast to antifungal agents. Moreover, oral cavity can act as a reservoir of certain pathogens that may cause systemic infections. The periodontal pocket is an ecological niche suitable to host microorganisms that could act as opportunistic pathogens. The aim of this study is to contribute to the understanding of resistance to conventional antifungal against C. albicans isolates from patients with periodontitis and diabetes. The determination of the minimal inhibitory concentrations (MIC) was evaluated according to M27S3 of the CLSI (2008), with modifications. The results showed that 48.8% of the studied strains were resistant to one or more antifungals and 6.6% were resistant to fluconazole and voriconazole. These results suggest an increasing resistance to conventional antifungal agents among Candida species, suggesting that the oral cavity could host pathogen fungi.