Self-cured resin modified by quaternary ammonium methacrylates and chlorhexidine: Cytotoxicity, antimicrobial, physical, and mechanical properties.

OBJECTIVE: To evaluate the addition of dimethylaminohexadecyl methacrylate (DMAHDM) and chlorhexidine diacetate on cytotoxicity, antimicrobial activity, physical, and mechanical properties of a self-cured resin. METHODS: 132 disk-shaped and 48 rectangular specimens were divided into four experimental groups as described: Control Group (CG - no addition), dCHX (1%), DMAHDM (5%), and DMAHDM+dCHX (5%+1%). The biofilm viability, flexural strength (FS - ISO 20795-1:2013), surface roughness (SR), and color stability (ΔE) were analyzed after being stored for 4 weeks in distilled water and immersed for 72h in coffee. Cytotoxicity was measured after 24h, 3, and 7 days of elution using an MTT test on L929 cells (ISO 10993-5:2009). SR and ΔE were measured by a contact profilometer and a spectrophotometer using the CIELab parameter. Data were submitted to ANOVA and Bonferroni's/Tukey's tests (p≤0.05). RESULTS: Significant antimicrobial activity against Streptococcus mutans and Candida albicans was detected in all groups when compared to the CG (p<0.05). Only the dCHX group, in 24h of elution, demonstrated no cytotoxicity effects. There was a statistical difference for FS on the tested groups (p<0.05). No differences were detected in the initial roughness' measurements among the groups (p>0.05). However, after storage and immersion in coffee, the groups containing DMAHDM presented with rougher surfaces and significantly lower color stability compared to the control (p<0.05). SIGNIFICANCE: The addition of dCHX and DMAHDM in self-cured resin presented antimicrobial properties; however, cytotoxicity, physical, and mechanical properties were compromised.

Bloodstream and catheter-related infections due to different clones of multidrug-resistant and biofilm producer Corynebacterium striatum.

BACKGROUND: Corynebacterium striatum is an emerging multidrug-resistant (MDR) pathogen associated with immunocompromised and chronically ill patients, as well as nosocomial outbreaks. In this study, we characterized 23 MDR C. striatum isolated of bloodstream and catheter-related infections from a hospital of Rio de Janeiro. METHODS: C. striatum isolates were identified by 16S rRNA and rpoB genes sequencing. The dissemination of these isolates was accomplished by pulsed-field gel electrophoresis (PFGE). All isolates were submitted to antimicrobial susceptibility testing by disk diffusion and by minimum inhibitory concentration using E-test strips methods. Antimicrobial resistance genes were detected by polymerase chain reaction. Quantitative tests were performed on four different abiotic surfaces and the ability to produce biofilm on the surface of polyurethane and silicone catheter was also demonstrated by scanning electron microscopy. RESULTS: Eleven PFGE profiles were found. The PFGE profile I was the most frequently observed among isolates. Five different MDR profiles were found and all PFGE profile I isolates presented susceptibility only to tetracycline, vancomycin, linezolid and daptomycin. Only the multidrug-susceptible isolate did not show mutations in the quinolone-resistance determinant region (QRDR) of the gyrA gene and was negative in the search of genes encoding antibiotic resistance. The other 22 isolates were positive to resistance genes to aminoglycoside, macrolides/lincosamides and chloramphenicol and showed mutations in the QRDR of the gyrA gene. Scanning electron microscopy illustrated the ability of MDR blood isolate partaker of the epidemic clone (PFGE profile I) to produce mature biofilm on the surface of polyurethane and silicone catheter. CONCLUSIONS: Genotyping analysis by PFGE revealed the permanence of the MDR PFGE profile I in the nosocomial environment. Other new PFGE profiles emerged as etiologic agents of invasive infections. However, the MDR PFGE profile I was also found predominant among patients with hematogenic infections. The high level of multidrug resistance associated with biofilm formation capacity observed in MDR C. striatum is a case of concern.

Epidemiological and clinical profile of infective endocarditis at a Brazilian tertiary care center: an eight-year prospective study.

INTRODUCTION: Infective endocarditis (IE) is a systemic infectious disease requiring a multidisciplinary team for treatment. This study presents the epidemiological and clinical data of 73 cases of IE in Rio de Janeiro, Brazil. METHODS: This observational prospective cohort study of endocarditis patients during an eight-year study period described 73 episodes of IE in 70 patients (three had IE twice). Community-associated (CAIE) and healthcare-acquired infective endocarditis (HAIE) were diagnosed according to the modified Duke criteria. The collected data included demographic, epidemiologic, and clinical characteristics, including results of blood cultures, echocardiographic findings, surgical interventions, and outcome. RESULTS: Analysis of data from the eight-year study period and 73 cases (70 patients) of IE showed a mean age of 46 years (SD=2.5 years; 1-84 years) and that 65.7% were male patients. The prevalence of CAIE and HAIE was 32.9% and 67.1%, respectively. Staphylococcus aureus (30.1%), Enterococcus spp. (19.1%), and Streptococcus spp. (15.0%) were the prevalent microorganisms. The relevant signals and symptoms were fever (97.2%; mean 38.6 + 0.05°C) and heart murmur (87.6%). Vegetations were observed in the mitral (41.1%) and aortic (27.4%) valves. The mortality rate of the cases was 47.9%. CONCLUSIONS: In multivariate analysis, chronic renal failure (relative risk [RR]= 1.60; 95% confidence interval [CI] 1.01-2.55), septic shock (RR= 2.19; 95% CI 1.499-3.22), and age over 60 years (RR= 2.28; 95% CI 1.44-3.59) were indirectly associated with in-hospital mortality. The best prognosis was related to the performance of cardiovascular surgery (hazard ratio [HR]= 0.51; 95% CI 0.26-0.99).

Evaluation of polymethyl methacrylate containing chlorhexidine: A randomized, controlled, split-mouth in situ study.

PURPOSE: To evaluate the antimicrobial action and elemental composition of chlorhexidine (CHX) diacetate in acrylic resins based on PMMA (polymethyl methacrylate) in situ. In addition, ex vivo evaluation of the CHX release mechanism was performed over a 14-day period. METHODS: Three discs of PMMA incorporating CHX and three control discs were mounted on individual oral splints and exposed to the oral cavity of 32 participants for 24 hours. The antimicrobial activity was evaluated by the plate count method. In the second test, elemental analysis of the specimens (n = 10) was performed by X-ray fluorescence before and after use of the device. Chlorhexidine release over a 14-day period was evaluated ex vivo in saliva samples collected from five individuals through proton nuclear magnetic resonance spectroscopy ( ¹H NMR) (500 MHz). RESULTS: Bacterial adhesion, evaluated by the plate count method, did not differ between the experimental material and control. (P> 0.05) The presence of the CHX molecule was detected by X-ray fluorescence before and after insertion of discs containing CHX into the oral cavity of participants. With regard to release, CHX was detected in saliva samples for 14 days and highest during the first 24 hours. When partial least squares discriminant analysis (PLS-DA) was applied in ¹H NMR, we observed a greater difference between the test and control groups. CLINICAL SIGNIFICANCE: The sustained release of CHX from PMMA suggests that such materials may be convenient for reducing the development of biofilm on the surface of the material for use in dentures and temporary restorative materials.

Epidemiological and clinical profile of infective endocarditis at a Brazilian tertiary care center: an eight-year prospective study

Abstract INTRODUCTION: Infective endocarditis (IE) is a systemic infectious disease requiring a multidisciplinary team for treatment. This study presents the epidemiological and clinical data of 73 cases of IE in Rio de Janeiro, Brazil. METHODS This observational prospective cohort study of endocarditis patients during an eight-year study period described 73 episodes of IE in 70 patients (three had IE twice). Community-associated (CAIE) and healthcare-acquired infective endocarditis (HAIE) were diagnosed according to the modified Duke criteria. The collected data included demographic, epidemiologic, and clinical characteristics, including results of blood cultures, echocardiographic findings, surgical interventions, and outcome. RESULTS: Analysis of data from the eight-year study period and 73 cases (70 patients) of IE showed a mean age of 46 years (SD=2.5 years; 1-84 years) and that 65.7% were male patients. The prevalence of CAIE and HAIE was 32.9% and 67.1%, respectively. Staphylococcus aureus (30.1%), Enterococcus spp. (19.1%), and Streptococcus spp. (15.0%) were the prevalent microorganisms. The relevant signals and symptoms were fever (97.2%; mean 38.6 + 0.05°C) and heart murmur (87.6%). Vegetations were observed in the mitral (41.1%) and aortic (27.4%) valves. The mortality rate of the cases was 47.9%. CONCLUSIONS: In multivariate analysis, chronic renal failure (relative risk [RR]= 1.60; 95% confidence interval [CI] 1.01-2.55), septic shock (RR= 2.19; 95% CI 1.499-3.22), and age over 60 years (RR= 2.28; 95% CI 1.44-3.59) were indirectly associated with in-hospital mortality. The best prognosis was related to the performance of cardiovascular surgery (hazard ratio [HR]= 0.51; 95% CI 0.26-0.99).

Genome sequence of a multidrug-resistant Corynebacterium striatum isolated from bloodstream infection from a nosocomial outbreak in Rio de Janeiro, Brazil.

Multidrug-resistant (MDR) Corynebacterium striatum has been cited with increased frequency as pathogen of nosocomial infections. In this study, we report the draft genome of a C. striatum isolated from a patient with bloodstream infection in a hospital of Rio de Janeiro, Brazil. The isolate presented susceptibility only to tetracycline, vancomycin and linezolid. The detection of various antibiotic resistance genes is fully consistent with previously observed multidrug-resistant pattern in Corynebacterium spp. A large part of the pTP10 plasmid of MDR C. striatum M82B is present in the genome of our isolate. A SpaDEF cluster and seven arrays of CRISPR-Cas were found.

Genome sequence of a multidrug-resistant Corynebacterium striatum isolated from bloodstream infection from a nosocomial outbreak in Rio de Janeiro, Brazil

Multidrug-resistant (MDR) Corynebacterium striatum has been cited with increased frequency as pathogen of nosocomial infections. In this study, we report the draft genome of a C. striatum isolated from a patient with bloodstream infection in a hospital of Rio de Janeiro, Brazil. The isolate presented susceptibility only to tetracycline, vancomycin and linezolid. The detection of various antibiotic resistance genes is fully consistent with previously observed multidrug-resistant pattern in Corynebacterium spp. A large part of the pTP10 plasmid of MDR C. striatum M82B is present in the genome of our isolate. A SpaDEF cluster and seven arrays of CRISPR-Cas were found.

Improved sealing ability promoted by calcium silicate-based root canal sealers

Aim: This study investigated the sealing ability produced by AH Plus (Dentsply DeTrey, Konstaz, Germany), EndoSequence BC sealer (Brasseler USA, Savannah, GA, USA), GuttaFlow (Coltène/Whaledent, Altstätten, France) and MTA Fillapex (Angelus, Londrina, Brazil). Methdos: A total of forty-six single-root human canines were prepared and randomly divided into four experimental groups (n=10): MTA Fillapex, EndoSequence BC Sealer, AH Plus or GuttaFlow. Teeth with intact crowns served as negative controls (n=3) and teeth filled with only gutta-percha served as positive controls (n=3). Teeth were mounted in a two-chamber apparatus and exposed to Enterococcus faecalis. The number of days over a 60-days period was recorded for the appearance of turbidity in the lower chamber. Kaplan-Meier method was used to estimate the survival curves. The nonparametric log-rank test was used to compare the survival curves using a significance level of 0.05. Results: The results at the end of the observation time were further analyzed by chi-square testing. All positive controls leaked within 24 h, whereas none of the negative controls leaked after 60 days. EndoSequence BC Sealer and MTA Fillapex had significant less bacterial leakage (P<0.05) than the other tested sealers. No significant difference between AH Plus and GuttaFlow was observed (P>0.05). Conclusion: In conclusion, calcium-silicate based root canal sealers promoted improved sealing ability when compared to other endodontic sealers

Endocarditis due to Rhodotorula mucilaginosa in a kidney transplanted patient: case report and review of medical literature.

Introduction. Endocarditis caused by yeasts is currently an emerging cause of infective endocarditis and, when accompanied byfever of unknown origin, is more severe since interferes with proper diagnosis and endocarditis treatment. Case presentation. The Rio de Janeiro Infective Endocarditis Study Group reports a case of infectious endocarditis (IE) with negative blood cultures in a 45-year-old white female resident in Rio de Janeiro, Brazil, previously submitted to kidney transplantation. After diagnosis and intervention, the valve culture revealed Rhodotorula mucilaginosa. The clinical aspects and overview of endocarditis caused by Rhodotorula spp. demonstrated that R. muscilaginosa have been isolated from the last IE cases from kidney transplanted patients. Conclusion. Though most of the patients (in literature) recovered well from endocarditis caused by Rhodotorula spp., physicians must be aware for diagnosis of fungemia and fungal treatment in kidney transplanted patients suffering of fever of unknown origin in the modern immunosuppressive treatment.

Cytotoxicity of three light-cured resin cements on 3T3 fibroblasts / Citotoxicidade de três cimentos resinosos fotopolimerizáveis sobre fibroblastos 3T3

Introduction: Light-cured resin cements are the first choice for the cementation of laminate veneers. Ideally, they should be biocompatible and offer minimum risks to patients. Objective: The aim of this study was to evaluate, in vitro, the cytotoxicity of three resin cements: Variolink II, Ivoclar Vivadent (C1), Allcem Veneer, FGM (C2), and Rely X Veneer, 3M ESPE (C3). Material and method: Twenty four samples of each of the cements were fabricated in a standardized metal mold, light activated, and transferred to a 96-well cell plate with culture of fibroblasts. After 24, 48, and 72h of incubation, cytotoxicity was assessed and cell viability was calculated by the methyl-thiazol-tetrazolium (MTT) colorimetric assay. Absorbance was measured at 570 nm using a microplate spectrophotometer. Result: The following results were found: Variolink II presented viability of 72.24% (SD 6.80) after 24h, 83.92% (SD 5.26) after 48h, and 92.77% (SD 5.59) after 72h; Allcem Veneer exhibited viability of 70.46% (SD 12.91) after 24h, 85.03% (SD 21.4) after 48h, and 70.46% (SD 12.91) after 72h; Rely X Veneer showed viability of 5.06% (SD 0.88) after 24h, 5.84% (SD 1.18) after 48h, and 6.99% (SD 1.34) after 72h. Conclusion: Under these testing conditions, Rely X Veneer presented significantly higher cytotoxicity compared with those of the other light-cured resin cements assessed.

Introdução: Cimentos resinosos fotopolimerizáveis são materiais de eleição para a cimentação de facetas laminadas. Devem ser biocompatíveis oferencendo riscos mínimos ao uso clínico em pacientes. Objetivo: O objetivo desse trabalho foi avaliar in vitro a citotoxicidade de três cimentos resinosos: Variolink II (Ivoclar Vivadent), Allcem Veneer, (FGM) e Rely X Veneer (3M ESPE). Material e método: Vinte e quatro corpos de prova de cada cimento foram confeccionados em matrizes metálicas padronizadas e inseridos em placa de cultura de células de noventa e seis poços contendo fibroblastos da linhagem 3T3. As células foram cultivadas em meio de cultivo celular RPMI 1640 com 5% de soro fetal bovino, com 0,1% de penicilina/estreptomicina em estufa a 37◦C, em atmosfera úmida com 5% de CO2. O grau de citotoxicidade de cada cimento foi avaliado após os tempos de contato de 24h, 48h e 72h através do método MTT (3-(4,5-dimetiltiazol-2yl)-2,5- difenil brometo de tetrazolina), que avalia a viabilidade celular pela função mitocondrial. Após os tempos estabelecidos, as amostras foram removidas, tratadas e levadas ao espectofotômetro de microplaca para leitura da absorbância em 570nm. Resultado: O cimento Variolink apresentou em 24h viabilidade de 72,24% (±6,80), em 48h de 83,92% (± 5,26) e de 92,77% (±5,59) em 72h. Allcem Veneer apresentou viabilidade de 70,46% (± 12,91) em 24h; 85,03% (± 21,4) em 48h e 70,46% (± 12,91) em 72h. O RelyX Veneer demonstrou viabilidade de 5,06% (± 0,88) em 24h; 5,84% (± 1,18) em 48h e 6,99% (± 1,34) em 72h. Conclusão: Estes resultados demonstraram que o cimento Rely-X se apresentou significativamente mais citotóxico nas condições testadas.

Sphygmomanometers and thermometers as potential fomites of Staphylococcus haemolyticus: biofilm formation in the presence of antibiotics

BACKGROUND The association between Staphylococcus haemolyticus and severe nosocomial infections is increasing. However, the extent to which fomites contribute to the dissemination of this pathogen through patients and hospital wards remains unknown. OBJECTIVES In the present study, sphygmomanometers and thermometers were evaluated as potential fomites of oxacillin-resistant S. haemolyticus (ORSH). The influence of oxacillin and vancomycin on biofilm formation by ORSH strains isolated from fomites was also investigated. METHODS The presence of ORSH on swabs taken from fomite surfaces in a Brazilian hospital was assessed using standard microbiological procedures. Antibiotic susceptibility profiles were determined by the disk diffusion method, and clonal distribution was assessed in pulsed-field gel electrophoresis (PFGE) assays. Minimum inhibitory concentrations (MICs) of oxacillin and vancomycin were evaluated via the broth microdilution method. Polymerase chain reaction (PCR) assays were performed to detect the mecA and icaAD genes. ORSH strains grown in media containing 1/4 MIC of vancomycin or oxacillin were investigated for slime production and biofilm formation on glass, polystyrene and polyurethane catheter surfaces. FINDINGS ORSH strains comprising five distinct PFGE types were isolated from sphygmomanometers (n = 5) and a thermometer (n = 1) used in intensive care units and surgical wards. ORSH strains isolated from fomites showed susceptibility to only linezolid and vancomycin and were characterised as multi-drug resistant (MDR). Slime production, biofilm formation and the survival of sessile bacteria differed and were independent of the presence of the icaAD and mecA genes, PFGE type and subtype. Vancomycin and oxacillin did not inhibit biofilm formation by vancomycin-susceptible ORSH strains on abiotic surfaces, including on the catheter surface. Enhanced biofilm formation was observed in some situations. Moreover, a sub-lethal dose of vancomycin induced biofilm formation by an ORSH strain on polystyrene. MAIN CONCLUSIONS Sphygmomanometers and thermometers are fomites for the transmission of ORSH. A sub-lethal dose of vancomycin may favor biofilm formation by ORSH on fomites and catheter surfaces.

Efetividade do ácido peracético na desinfecção rápida de cones de guta-percha e de Resilon expostos ao Enterococcus faecalis / Effectiveness of peracetic acid in rapid disinfection of gutta-percha and Resilon cones exposed to Enterococcus faecalis

Objetivo: o objetivo desse trabalho foi verificar a efetividade do ácido peracético a 2% na descontaminação rápida de cones de guta-percha e de Resilon, comparado ao hipoclorito de sódio e à clorexidina. Métodos: os cones de guta-percha e Resilon foram imersos por cinco minutos para contaminação em suspensão de Enterococcus faecalis, e divididos em grupos (n = 10): ácido peracético a 2% (um e três minutos); NaOCl a 5,25% (um e três minutos); clorexidina a 2% (um e três minutos). Após a realização dos protocolos em teste, os cones foram transferidos para tubos de ensaio contendo meio Enterococcosel® e, então, foram mantidos em estufa a 37º por 48 horas. Após o período de observação, os tubos foram avaliados e os que apresentam turvação do meio foram consideradas positivos. Resultados: os resultados demonstraram que o ácido peracético a 2% parece ser efetivo para a descontaminação de ambos os tipos de cone, de forma alternativa ao NaOCl a 5,25%, enquanto a clorexidina a 2% apresentou menor efetividade (p < 0,05). Conclusão: Ambas as soluções, ácido peracético a 2% e NaOCl a 5,25%, foram efetivas na descontaminação de cones de guta-percha ou de Resilon nos tempos testados.

Tellurite resistance: a putative pitfall in Corynebacterium diphtheriae diagnosis?

Corynebacterium diphtheriae strains continue to circulate worldwide causing diphtheria and invasive diseases, such as endocarditis, osteomyelitis, pneumonia and catheter-related infections. Presumptive C. diphtheriae infections diagnosis in a clinical microbiology laboratory requires a primary isolation consisting of a bacterial culture on blood agar and agar containing tellurite (TeO3(2-)). In this study, nine genome sequenced and four unsequenced strains of C. diphtheriae from different sources, including three samples from a recent outbreak in Brazil, were characterized with respect to their growth properties on tellurite-containing agar. Levels of tellurite-resistance (Te(R)) were evaluated by determining the minimum inhibitory concentrations of potassium tellurite (K2TeO3) and by a viability reduction test in solid culture medium with K2TeO3. Significant differences in Te(R) levels of C. diphtheriae strains were observed independent of origin, biovar or presence of the tox gene. Data indicated that the standard initial screening with TeO3(2-)-selective medium for diphtheria bacilli identification may lead to false-negative results in C. diphtheriae diagnosis laboratories.

Corynebacterium diphtheriae putative tellurite-resistance protein (CDCE8392_0813) contributes to the intracellular survival in human epithelial cells and lethality of Caenorhabditis elegans

Corynebacterium diphtheriae, the aetiologic agent of diphtheria, also represents a global medical challenge because of the existence of invasive strains as causative agents of systemic infections. Although tellurite (TeO3^2-) is toxic to most microorganisms, TeO3^2--resistant bacteria, including C. diphtheriae, exist in nature. The presence of TeO3^2--resistance (Te^R) determinants in pathogenic bacteria might provide selective advantages in the natural environment. In the present study, we investigated the role of the putative Te^R determinant (CDCE8392_813gene) in the virulence attributes of diphtheria bacilli. The disruption of CDCE8392_0813 gene expression in the LDCIC-L1 mutant increased susceptibility to TeO3^2- and reactive oxygen species (hydrogen peroxide), but not to other antimicrobial agents. The LDCIC-L1 mutant also showed a decrease in both the lethality of Caenorhabditis elegansand the survival inside of human epithelial cells compared to wild-type strain. Conversely, the haemagglutinating activity and adherence to and formation of biofilms on different abiotic surfaces were not regulated through the CDCE8392_0813 gene. In conclusion, the CDCE8392_813 gene contributes to the Te^R and pathogenic potential of C. diphtheriae.

Corynebacterium diphtheriae putative tellurite-resistance protein (CDCE8392_0813) contributes to the intracellular survival in human epithelial cells and lethality of Caenorhabditis elegans.

Corynebacterium diphtheriae, the aetiologic agent of diphtheria, also represents a global medical challenge because of the existence of invasive strains as causative agents of systemic infections. Although tellurite (TeO32-) is toxic to most microorganisms, TeO32--resistant bacteria, including C. diphtheriae, exist in nature. The presence of TeO32--resistance (TeR) determinants in pathogenic bacteria might provide selective advantages in the natural environment. In the present study, we investigated the role of the putative TeR determinant (CDCE8392_813gene) in the virulence attributes of diphtheria bacilli. The disruption of CDCE8392_0813 gene expression in the LDCIC-L1 mutant increased susceptibility to TeO32- and reactive oxygen species (hydrogen peroxide), but not to other antimicrobial agents. The LDCIC-L1 mutant also showed a decrease in both the lethality of Caenorhabditis elegans and the survival inside of human epithelial cells compared to wild-type strain. Conversely, the haemagglutinating activity and adherence to and formation of biofilms on different abiotic surfaces were not regulated through the CDCE8392_0813 gene. In conclusion, the CDCE8392_813 gene contributes to the TeR and pathogenic potential of C. diphtheriae.

Biofilm production by multiresistant Corynebacterium striatum associated with nosocomial outbreak.

Corynebacterium striatum is a potentially pathogenic microorganism that causes nosocomial outbreaks. However, little is known about its virulence factors that may contribute to healthcare-associated infections (HAIs). We investigated the biofilm production on abiotic surfaces of multidrug-resistant (MDR) and multidrug-susceptible (MDS) strains of C. striatum of pulsed-field gel electrophoresis types I-MDR, II-MDR, III-MDS and IV-MDS isolated during a nosocomial outbreak in Rio de Janeiro, Brazil. The results showed that C. striatum was able to adhere to hydrophilic and hydrophobic abiotic surfaces. The C. striatum 1987/I-MDR strain, predominantly isolated from patients undergoing endotracheal intubation procedures, showed the greatest ability to adhere to all surfaces. C. striatum bound fibrinogen to its surface, which contributed to biofilm formation. Scanning electron microscopy showed the production of mature biofilms on polyurethane catheters by all pulsotypes. In conclusion, biofilm production may contribute to the establishment of HAIs caused by C. striatum.

Biofilm formation and fibrinogen and fibronectin binding activities by Corynebacterium pseudodiphtheriticum invasive strains.

Biofilm-related infections are considered a major cause of morbidity and mortality in hospital environments. Biofilms allow microorganisms to exchange genetic material and to become persistent colonizers and/or multiresistant to antibiotics. Corynebacterium pseudodiphtheriticum (CPS), a commensal bacterium that colonizes skin and mucosal sites has become progressively multiresistant and responsible for severe nosocomial infections. However, virulence factors of this emergent pathogen remain unclear. Herein, we report the adhesive properties and biofilm formation on hydrophilic (glass) and hydrophobic (plastic) abiotic surfaces by CPS strains isolated from patients with localized (ATCC10700/Pharyngitis) and systemic (HHC1507/Bacteremia) infections. Adherence to polystyrene attributed to hydrophobic interactions between bacterial cells and this negatively charged surface indicated the involvement of cell surface hydrophobicity in the initial stage of biofilm formation. Attached microorganisms multiplied and formed microcolonies that accumulated as multilayered cell clusters, a step that involved intercellular adhesion and synthesis of extracellular matrix molecules. Further growth led to the formation of dense bacterial aggregates embedded in the exopolymeric matrix surrounded by voids, typical of mature biofilms. Data also showed CPS recognizing human fibrinogen (Fbg) and fibronectin (Fn) and involvement of these sera components in formation of "conditioning films". These findings suggested that biofilm formation may be associated with the expression of different adhesins. CPS may form biofilms in vivo possibly by an adherent biofilm mode of growth in vitro currently demonstrated on hydrophilic and hydrophobic abiotic surfaces. The affinity to Fbg and Fn and the biofilm-forming ability may contribute to the establishment and dissemination of infection caused by CPS.

Biofilm production by multiresistant Corynebacterium striatum associated with nosocomial outbreak

Corynebacterium striatum is a potentially pathogenic microorganism that causes nosocomial outbreaks. However, little is known about its virulence factors that may contribute to healthcare-associated infections (HAIs). We investigated the biofilm production on abiotic surfaces of multidrug-resistant (MDR) and multidrug-susceptible (MDS) strains of C. striatum of pulsed-field gel electrophoresis types I-MDR, II-MDR, III-MDS and IV-MDS isolated during a nosocomial outbreak in Rio de Janeiro, Brazil. The results showed that C. striatum was able to adhere to hydrophilic and hydrophobic abiotic surfaces. The C. striatum 1987/I-MDR strain, predominantly isolated from patients undergoing endotracheal intubation procedures, showed the greatest ability to adhere to all surfaces. C. striatum bound fibrinogen to its surface, which contributed to biofilm formation. Scanning electron microscopy showed the production of mature biofilms on polyurethane catheters by all pulsotypes. In conclusion, biofilm production may contribute to the establishment of HAIs caused by C. striatum.

Efficacy of chemomechanical caries removal in reducing cariogenic microbiota: a randomized clinical trial.

The aim of this study was to compare the efficacy of chemochemical methods (Carisolv™ and Papacárie®) versus the manual method (excavators) in reducing the cariogenic microbiota in dentine caries of primary teeth. Forty-six healthy children (5 to 9 years old) having at least one primary tooth with a cavitated dentine carious lesion were included in the study. The teeth presented no clinical or radiographic signs of pulpal involvement. The sample of 74 teeth was randomly divided into three different groups: Papacárie® (n = 25), Carisolv™ (n = 27) and Manual (n = 22). Samples of carious and sound dentine were collected with sterile excavators before and after caries removal in the three groups. The dentine samples were transferred to glass tubes containing a 1mL thioglycollate medium used as a carrier and enriched for microbiological detection of mutans streptococci and Lactobacillus spp, after incubation for 6h at room temperature. The minimum detection value for colony forming units (CFU) was 3.3 x 102 CFU/ml, and the results were converted into scores from 0 to 4. A significant difference was observed in relation to the microbiological scores before and after caries removal for all methods (Wilcoxon test; p < 0.001). The use of chemomechanical methods for caries removal did not improve the reduction of cariogenic microorganisms in dentine caries lesions, in comparison with manual excavation.

Molecular identification of nocardia isolates from clinical samples and an overview of human nocardiosis in Brazil.

BACKGROUND: Nocardia sp. causes a variety of clinical presentations. The incidence of nocardiosis varies geographically according to several factors, such as the prevalence of HIV infections, transplants, neoplastic and rheumatic diseases, as well as climate, socio-economic conditions and laboratory procedures for Nocardia detection and identification. In Brazil the paucity of clinical reports of Nocardia infections suggests that this genus may be underestimated as a cause of human diseases and/or either neglected or misidentified in laboratory specimens. Accurate identification of Nocardia species has become increasingly important for clinical and epidemiological investigations. In this study, seven clinical Nocardia isolates were identified by multilocus sequence analysis (MLSA) and their antimicrobial susceptibility was also determined. Most Nocardia isolates were associated to pulmonary disease. METHODOLOGY/PRINCIPAL FINDINGS: The majority of Brazilian human isolates in cases reported in literature were identified as Nocardia sp. Molecular characterization was used for species identification of Nocardia nova, Nocardia cyriacigeorgica, Nocardia asiatica and Nocardia exalbida/gamkensis. Data indicated that molecular analysis provided a different Nocardia speciation than the initial biochemical identification for most Brazilian isolates. All Nocardia isolates showed susceptibility to trimethoprim-sulfamethoxazole, the antimicrobial of choice in the treatment nocardiosis. N. nova isolated from different clinical specimens from one patient showed identical antimicrobial susceptibility patterns and two distinct clones. CONCLUSIONS/SIGNIFICANCE: Although Brazil is the world's fifth-largest country in terms of land mass and population, pulmonary, extrapulmonary and systemic forms of nocardiosis were reported in only 6 of the 26 Brazilian states from 1970 to 2013. A least 33.8% of these 46 cases of nocardiosis proved fatal. Interestingly, coinfection by two clones may occur in patients presenting nocardiosis. Nocardia infection may be more common throughout the Brazilian territory and in other developing tropical countries than is currently recognized and MLSA should be used more extensively as an effective method for Nocardia identification.