Raphael Hirata Junior

Diphtheria remains a threat to health in the developing world: an overview

Changes in the epidemiology of diphtheria are occurring worldwide. A large proportion of adults in many industrialized and developing countries are now susceptible to diphtheria. Vaccine-induced immunity wanes over time unless periodic booster is given or exposure to toxigenic Corynebacterium diphtheriae occurs. Immunity gap in adults coupled with large numbers of susceptible children creates the potential for new extensive epidemics. Epidemic emergencies may not be long in coming in countries experiencing rapid industrialization or undergoing sociopolitical instability where many of the factors thought to be important in producing epidemic such as mass population movements and difficult hygienic and economic conditions are present. The continuous circulation of toxigenic C. diphtheriae emphasizes the need to be aware of epidemiological features, clinical signs, and symptoms of diphtheria in vaccine era so that cases can be promptly diagnosed and treated, and further public health measures can be taken to contain this serious disease. This overview focused on worldwide data obtained from diphtheria with particular emphasis to main factors leading to recent epidemics, new clinical forms of C. diphtheriae infections, expression of virulence factors, other than toxin production, control strategies, and laboratory diagnosis procedures.

Antimicrobial activity of different concentrations of NaOCl and chlorhexidine using a contact test

The purpose of this study was to analyze the in vitro antimicrobial activity of sodium hypochlorite (1% and 5%) and chlorhexidine (0.12%, 0.5% and 1%). Bacterial samples (ATCC) of Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Porphyromonas gingivalis and Fusobacterium nucleatum were submitted to a contact test. Solutions were evaluated at different time intervals: immediately, 5 min, 15 min, and 30 min after contact and repeated 10 times. The results of the contact test showed that 0.12% chlorhexidine did not eliminate E. faecalis at any time interval, while 0.5% and 1% chlorhexidine and 1% and 5% sodium hypochlorite did. These results permit us to conclude that to obtain better antimicrobial activity, chlorhexidine in a concentration greater than 0.12% should be used.

Corynebacterium ulcerans diphtheria: an emerging zoonosis in Brazil and worldwide

The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America.O artigo revisa a literatura sobre a emergencia de infeccoes humanas causadas por Corynebacterium ulcerans em diversos paises, incluindo o Brasil. Foi realizada analise de artigos publicados entre 1926 e 2011 nas bases Medline/PubMed e SciELO, bem como artigos e informes do Ministerio da Saude. Apresenta-se um esquema de triagem, rapido, economico e de facil execucao, capaz de permitir a realizacao do diagnostico presuntivo de C. ulcerans e C. diphtheriae na maioria dos laboratorios brasileiros publicos e privados. A circulacao de C. ulcerans em varios paises, aliada aos recentes casos de isolamento do patogeno no Rio de Janeiro, e um alerta a clinicos, veterinarios e microbiologistas sobre a ocorrencia de difteria zoonotica e a circulacao do C. ulcerans em regioes urbanas e rurais do territorio nacional e/ou da America Latina.

Corynebacterium ulcerans isolated from an asymptomatic dog kept in an animal shelter in the metropolitan area of Rio de Janeiro, Brazil.

Corynebacterium ulcerans was isolated from nares of one asymptomatic dog kept in an animal shelter in the metropolitan area of Rio de Janeiro, Brazil. The RNA polymerase beta subunit-encoding gene was sequenced to confirm the species identity. C. ulcerans strains producing phospholipase D, but not diphtheria toxin, are able to cause severe disease in humans, such as pneumonia and granulomatous nodules in pulmonary tissues. The infection rate varies really widely by region, probably because of the variations in the reported infection rates. Dogs with unapparent C. ulcerans infections may be considered as potentially capable of infecting other animals and humans, including pet owners. Medical and veterinary staff should be aware that asymptomatic animals can carry C. ulcerans and cooperate in eliminating infections and monitoring animals also in the developing countries.

Antimicrobial activity of calcium hydroxide pastes on Enterococcus faecalis cultivated in Root Canal Systems

The effectiveness of calcium hydroxide pastes: Calen and PMCC-Calen associated to chemo-mechanical preparation was assessed on Enterococcus faecalis grown within root canals. Seventy incisors were inserted into TSB medium, sterilized and contaminated with E. faecalis. Culture medium was replaced each 24 h and incubated at 37 degrees C for 72 h. After chemo-mechanical preparation, root canals were filled with Calen or PMCC-Calen (7 or 14 days). Pastes were removed and teeth were inserted into test tubes containing Enterococcosel broth. Calen paste (maintained for 7 and 14 days) induced 70% elimination of enterococci and PMCC-Calen 100% elimination only after maintenance for 14 days. These medications were significantly more effective (p<0.001) than chemo-mechanical protocol alone and PMCC-Calen maintained for 7 days, both incapable to eliminate the viability of enterococci. Calcium hydroxide pastes demonstrated important adjuvant effects in the elimination of enterococci during chemo-mechanical preparation of root canal systems. When associated with PMCC, calcium hydroxide pastes should be maintained for at least 14 days.

Antimicrobial resistance among Brazilian Corynebacterium diphtheriae strains

The increasing problems with multidrug resistance in relation to Corynebacterium, including C. diphtheriae, are examples of challenges confronting many countries. For this reason, Brazilian C. diphtheriae strains were evaluated by the E-Test for their susceptibility to nine antibacterial drugs used in therapy. Resistance (MIC < 0.002; 0.38 microg/ml) to penicillin G was found in 14.8% of the strains tested. Although erythromycin (MIC90 0.75 microg/ml) and azithromycin (MIC90 0.064 microg/ml) were active against C. diphtheriae in this study, 4.2% of the strains showed decreased susceptibility (MIC 1.0 microg/ml) to erythromycin. Multiple resistance profiles were determined by the disk diffusion method using 31 antibiotics. Most C. diphtheriae strains (95.74%) showed resistance to mupirocin, aztreonam, ceftazidime, and/or oxacillin, ampicillin, penicillin, tetracycline, clindamycin, lincomycin, and erythromycin. This study presents the antimicrobial susceptibility profiles of Brazilian C. diphtheriae isolates. The data are of value to practitioners, and suggest that some concern exists regarding the use of penicillin.

Antimicrobial resistance and virulence traits of Enterococcus faecalis from primary endodontic infections

OBJECTIVES To determine the phenotypic and molecular characteristics of Enterococcus faecalis recovered from primary endodontic infections in Brazilian patients. METHODS Twenty isolates of E. faecalis recovered from 43 Brazilian patients with primary endodontic infections were identified by biochemical profiling (API20Strep) and 16S rDNA sequencing. Antimicrobial susceptibility was ascertained by agar dilution, using the recommended protocol of the Clinical and Laboratory Standards Institute (CLSI). PCR with validated primers was used to detect genes associated with antibiotic resistance and specific virulence factors. RESULTS All isolates were deemed susceptible to penicillin G, erythromycin and vancomycin. However, nine isolates had a minimum inhibitory concentration of 4μg/mL to vancomycin (the resistance breakpoint). Fourteen isolates (70% of isolates) were also resistant to tetracycline with MICs of >64μg/mL. PCR products for tetracycline resistance genes were detected in test isolates, while erythromycin and vancomycin resistance genes were not evident. Gelatinase, aggregation substance and enteroccocal surface protein genes were detected in 20, 18 and 12 isolates, respectively. CONCLUSIONS Endodontic E. faecalis isolates exhibit high level of resistance to tetracycline, an antibiotic that has use in local treatment of dental infections. This opens up a much-needed debate on the role and efficacy of this antibiotic for oral infections. Furthermore, these isolates were shown to possess genes that could contribute to pathogenicity in the pulp cavity.

Difteria pelo Corynebacterium ulcerans: uma zoonose emergente no Brasil e no mundo

The article is a literature review on the emergence of human infections caused by Corynebacterium ulcerans in many countries including Brazil. Articles in Medline/PubMed and SciELO databases published between 1926 and 2011 were reviewed, as well as articles and reports of the Brazilian Ministry of Health. It is presented a fast, cost-effective and easy to perform screening test for the presumptive diagnosis of C. ulcerans and C. diphtheriae infections in most Brazilian public and private laboratories. C. ulcerans spread in many countries and recent isolation of this pathogen in Rio de Janeiro, southeastern Brazil, is a warning to clinicians, veterinarians, and microbiologists on the occurrence of zoonotic diphtheria and C. ulcerans dissemination in urban and rural areas of Brazil and/or Latin America.O artigo revisa a literatura sobre a emergencia de infeccoes humanas causadas por Corynebacterium ulcerans em diversos paises, incluindo o Brasil. Foi realizada analise de artigos publicados entre 1926 e 2011 nas bases Medline/PubMed e SciELO, bem como artigos e informes do Ministerio da Saude. Apresenta-se um esquema de triagem, rapido, economico e de facil execucao, capaz de permitir a realizacao do diagnostico presuntivo de C. ulcerans e C. diphtheriae na maioria dos laboratorios brasileiros publicos e privados. A circulacao de C. ulcerans em varios paises, aliada aos recentes casos de isolamento do patogeno no Rio de Janeiro, e um alerta a clinicos, veterinarios e microbiologistas sobre a ocorrencia de difteria zoonotica e a circulacao do C. ulcerans em regioes urbanas e rurais do territorio nacional e/ou da America Latina.

Clonal multidrug-resistant Corynebacterium striatum within a nosocomial environment, Rio de Janeiro, Brazil

Corynebacterium striatum is a potentially pathogenic microorganism with the ability to produce outbreaks of nosocomial infections. Here, we document a nosocomial outbreak caused by multidrug-resistant (MDR) C. striatum in Rio de Janeiro, Brazil. C. striatum identification was confirmed by 16S rRNA and rpoB gene sequencing. Fifteen C. striatum strains were isolated from adults (half of whom were 50 years of age and older). C. striatum was mostly isolated in pure culture from tracheal aspirates of patients undergoing endotracheal intubation procedures. The analysis by pulsed-field gel electrophoresis (PFGE) indicated the presence of four PFGE profiles, including two related clones of MDR strains (PFGE I and II). The data demonstrated the predominance of PFGE type I, comprising 11 MDR isolates that were mostly isolated from intensive care units and surgical wards. A potential causal link between death and MDR C. striatum (PFGE types I and II) infection was observed in five cases.

Comparative evaluation of antimicrobial action of MTA, calcium hydroxide and Portland cement

The present study aimed to evaluate and compare the antimicrobial effect of MTA Dentsply, MTA Angelus, Calcium Hydroxide and Portland cement. Four reference bacterial strains were used: Pseudomonas aeruginosa, Escherichia coli, Bacteroides fragilis, and Enterococcus faecalis. Plates containing Mueller-Hinton agar supplemented with 5% sheep blood, hemin, and menadione were inoculated with the bacterial suspensions. Subsequently, wells were prepared and immediately filled with materials and incubated at 37°C for 48 hours under anaerobic conditions, except P. aeruginosa. The diameters of inhibition zones were measured, and data analyzed using ANOVA and the Tukey test with 1% level of significance. MTA Dentsply, MTA Angelus and Portland cement inhibited the growth of P. aeruginosa. Calcium Hydroxide was effective against P. aeruginosa and B. fragillis. Under anaerobic conditions, which may hamper the formation of reactive oxygen species, the materials failed to inhibit E. faecalis, and E. coli.