Evaluation of biofilm formation on acrylic resin surfaces coated with silicon dioxide: an in situ study

Abstract Biofilm on acrylic resin dental prostheses may cause gingival inflammation. This study evaluated the influence of a silicon dioxide coating layer applied onto acrylic resin on the adhesion of microorganisms. Blocks (5 x 5 x 3 mm) of acrylic resin were evaluated for surface roughness and divided into two groups: control (CG) and coated with silicon dioxide (LG group). The specimens were evaluated by scanning electron microscopy (n = 1) and by contact angle analysis (n = 3). For the in situ study, 20 volunteers wore acrylic palatal devices containing three samples from each group (n = 60) for 2 days. The biofilm formed was quantified by metabolic activity and total biomass using the crystal violet assay. The results were subjected to Bartlett's normality test and Gamma model with random effect for the response variable (α = 5%). The mean contact angle of the coated group was significantly lower than that of the uncoated group (p < 0.05). The metabolic activity of microorganisms in the biofilm on the blocks treated with coating was significantly lower than that of control blocks (p = 0.02). Regarding the amount of extracellular matrix produced by the microorganisms, there was no difference between the CG and LG group (p = 0.05). The application of a silicon dioxide coating on acrylic resin reduced the activity of the polymicrobial biofilm formed in situ. This coating may be advantageous for patients with conventional complete dentures or implants made of acrylic resin and who have motor difficulties that prevent them from cleaning their prostheses properly.

Standardization of resazurin use in susceptibility testing of natural products against yeasts in planktonic cells and in biofilms formation

Natural products, such as the ethanolic propolis extract (PE), have been shown to be a safe and effective therapeutic alternative for the treatment of fungal skin and nail diseases. However, the presence of the resin and the physicochemical characteristics of the extract sometimes difficult the reading and determination of breakpoints of the in vitrotests, evidencing the need for alternatives that facilitate the reading. The present study aimed to standardize the use of resazurin in tests of susceptibility of PE with planktonic yeast cells and biofilm forms. The antifungal activity of PE was determined by minimum inhibitory concentration (MIC) and we observed that, for all Candida spp. tested, the most reproducible MIC results were obtained when resazurin was placed after 24 hours of incubation and remained more 24 hourswith yeasts plus PE. For encapsulated yeasts, there was no dye reduction and color transition. Resazurin was also used for the evaluation of minimal biofilm inhibitory concentration and minimal biofilm eradication concentration and it was metabolized and reproducedthe action of PE on Candida biofilms. In addition, microdilution checkerboard plates were made with the dye, which assisted reading the result of the interaction between PE and nystatin. We observed that the resin, the color and the turbidity of the PE slightly changed the color of the resazurin in high concentrations of the extract and did not impair the reading. Therefore, the resazurin standardization tests were proven to be efficient and grounds that it should be used as an auxiliary methodology for reading and interpretation of the susceptibility tests for non-encapsulated yeasts with natural products, which form turbidity or precipitation, such as propolis.

Fatores de virulência e susceptibilidade a antifúngicos de Cryptococcus Spp / Virulence factors and susceptibility to Cryptococcus Spp. antifungals

Criptococose é uma doença grave que afeta tanto imunocomprometidos quanto imunocompetentes, com isso analisar a virulência é fundamental para novas terapêuticas. Objetivo: Analisar a capacidade de virulência e susceptibilidade aos antifúngicos de Cryptococcus spp. isolados de líquor de pacientes de hospital do norte do Paraná. Métodos: A partir de dois isolados clínicos C. neoformans e C. gattii, realizou-se a confirmação da identificação. Para a virulência, avaliou-se o tamanho da cápsula, capacidade de sobrevivência após exposição a neutrófilos, produção de melanina e urease. No antifungigrama por difusão em disco utilizou-se: anfotericina B, cetoconazol, voriconazol, itraconazol e miconazol. Resultados: C. gattii destaca-se por maior desenvolvimento da cápsula além da melhor capacidade de sobreviver a fagocitose em relação ao C. neoformans. No antifungigrama, ambos os isolados se apresentam sensíveis às drogas estudadas. Conclusão: Esses achados contribuem para a compreensão das diferentes patogêneses entre C. gattii e C. neoformans.

Cryptococcosis is a serious disease that can affect both immunocompromised and immunocompetent individuals, thus the virulence analysis is fundamental for the development of new treatments. Objective: To analyze the virulence and susceptibility of Cryptococcus spp. isolated from cerebrospinal fluid of patients from a hospital in the north of Paraná. Methods: From two clinical isolates, C. neoformans and C. gattii were confirmed and identified. For virulence, capsule size, survival capacity after exposure to neutrophils, melanin production and urease were evaluated. In the disc-diffusion method, the following antifungals were used: amphotericin B, ketoconazole, voriconazole, itraconazole and miconazole Results: It was observed that C. gattii presents greater results for development of the capsule beside presenting the best ability to survive phagocytosis in relation to C. neoformans. In the disc-diffusion method, both isolates presented sensitivity to the studied drugs. Conclusion: These findings contribute to the understanding of the different pathogens between C. gattii and C. neoformans.

Effect of Silicon dioxide coating of acrylic resin surfaces on Candida albicans adhesion

Abstract Acrylic resin has been used in the manufacture of prostheses, however, in the oral cavity, this material starts to retain microorganisms capable of causing gingival inflammation due its porosities. The aim of this study was to evaluate the influence of the use of silicon dioxide as a coating layer applied onto acrylic resin, on the adhesion of Candida albicans (Ca). After the incubation period in Sabouraud Dextrose Broth, a total of 1 ml of the Ca suspension was added to plate wells, each well containing a specimen of acrylic resin. The adhesion ability of Ca on acrylic resin was determined by counting colonies. Three groups (n = 6) of acrylic resin were assessed: with polishing (RP); without polishing (RW); with polishing and coating layer of silicon dioxide (RPC). Ca deposited on the surface of the acrylic resin was also observed using Scanning Electron Microscopy (SEM). Statistical assessment by Kruskal-Wallis and Student-Newman-Keuls Method were done (α = 2%). There was significant difference among the groups. The RPC group showed the lowest growth, with an average of 5.59 Log CFU/cm 2 ; there was a statistically significant difference in relation to group RW, which presented a growth of 6.07 Log CFU/cm 2 and to group RP with 5.91 Log CFU/cm 2 (p < 000.1). SEM images demonstrated that in the RP and RPC group, the surface of the resin had greater regularity, and smaller number of microorganisms. The application of silicon dioxide coating on acrylic resin appears to be a promising alternative, and its use can help in reducing the adhesion of Ca in prostheses.

Occurrence of dermatophytoses in patients from the Sistema Único de Saúde

Abstract: Background: Dermatophytosis is a cutaneous disease caused by filamentous keratinophilic fungi belonging to the genera Trichophyton, Microsporum and Epidermophyton, which present a high prevalence in the general population, being among the most common mycoses affecting about 20% of the world's population. Objective: To carry out the epidemiological survey of cases of dermatophytosis in patients from the Sistema Único de Saúde in a regional Laboratory in the period of 5 years (2009 to 2013). Methods: A retrospective study (January 2009 to December 2013) was carried out with a qualitative and quantitative design, through the registry book of the laboratory, Mycology Sector, where cases of patients with suspected dermatomycosis were analyzed. Results: In a 5-year period, a total of 4467 cases were suspected of having a fungal infection. Of these, 68.74% (3071) cases were of dermatomycosis. In relation to cultures with fungal growth, 12.54% (385 cases) were dermatophyte fungi and 7.97% (245 cases) non-dermatophyte fungi were isolated. Among the species identified, there was a higher prevalence of T. rubrum complex (75%), T. mentagrophytes complex (11.68%) and M. canis (7.01%). Regarding the sites analyzed, nail involvement was the most frequent in 75% of the cases. Study Limitations: This work is representative in the studied region. Conclusions: Dermatomycosis samples are the most frequent among all samples of fungal infections from these patients, with the nail being the most affected area and the fungi T. rubrum complex and T. mentagrophytes complex the most frequent.