Pneumococcal nasopharyngeal carriage among children in Brazil prior to the introduction of the 10-valent conjugate vaccine: a culture- and PCR-based survey.

We performed two different approaches (broth enrichment step prior to culture (BEC) and PCR (BEPCR)) for detecting Streptococcus pneumoniae from nasopharyngeal specimens collected from 242 children aged <6 years attending one hospital (n = 140) and one childcare centre (n = 102) in a major urban area in Brazil. These specimens were collected immediately before the introduction of the 10-valent pneumococcal conjugate vaccine (PCV10) and the 13-valent vaccine (PCV13) for routine use in Brazil. Results were compared with previous findings obtained with direct culture (DC) on a selective medium. Colonisation prevalence was 58·3% (n = 141), being higher among children attending the childcare centre (62·7% vs. 55%). The culture-based methods (DC and BEC) enabled the detection of S. pneumoniae in 119 (49·2%) and 115 (47·5%) children, respectively. The PCR-based method (BEPCR) was more sensitive and 137 (56·6%) carriers were identified. Twenty-six serogroups/serotypes were identified, predominantly 6B, 19F, 14, 6A, 15C and 23F. Multiple colonisation was observed in 13 (5·4%) children. The estimated serotypes coverage of available PCVs was 40·4% for the 10-valent (included in the Brazilian immunisation programme) and 55·8% for the 13-valent (only available in private clinics). The use of robust approaches to obtain a more realistic insight about the asymptomatic carrier status is of paramount importance to estimate and assess the impact of vaccine implementation. The combination between culture-based and molecular methods constitutes a suitable strategy.

Prevalence and factors associated with anal lesions mediated by human papillomavirus in men with HIV/AIDS.

Prevalence and risk factors for human papillomavirus-related anal lesions were evaluated in 60 men with human immunodeficiency virus/acquired immunodeficiency syndrome (HIV/AIDS). Patients underwent anal cytology, anuscopy under colposcopic vision and anal biopsy for detection of the lesions. The mean age was 41.9 years and the mean time of HIV infection was 6.8 years, 88.3% of them having been on highly active antiretroviral therapy for an average of 6.5 years. Homosexuals represented 43.3% and bisexuals 15.0% whereas heterosexuals comprised 41.7%. The prevalence of anal lesions detected by anuscopy under colposcopic vision, cytology and biopsy were, respectively, 35.0, 16.7 and 23.3%. Homosexuals or bisexuals accounted for 85.7% of the patients with an abnormal biopsy, the remaining 14.3% being heterosexuals (P = 0.02). The T-CD(4)(+) lymphocyte count, HIV viral load and use of antiretrovirals did not reveal any association with anal lesions. The occurrence of anal lesions was high in the individuals with HIV/AIDS, especially in the homosexuals and bisexuals, but it also occurred in heterosexuals, justifying the screening of anal lesions of all men with the infection.

Brazilian paediatricians' awareness of national guidelines for community-acquired pneumonia.

BACKGROUND: The Integrated Management of Childhood Illness (IMCI) strategy uses simple clinical signs for the diagnosis and severity evaluation of community-acquired pneumonia (CAP). OBJECTIVE: To describe paediatrician awareness of the IMCI strategy for CAP. DESIGN: A cross-sectional study analysing a descriptive case of severe CAP, presented as part of the Brazilian Board of Paediatrician Evaluation (BBPE) tests. RESULTS: Eighty-six (774/898) per cent of paediatricians followed the IMCI protocol to treat CAP. Although hospitalisation was considered in 90% of the answers, only 35% based this decision on lower chest indrawing. CONCLUSION: The BBPE showed that most physicians are aware of the IMCI recommendations.

Antimicrobial susceptibility and survey of macrolide resistance mechanisms among Streptococcus pyogenes isolated in Rio de Janeiro, Brazil.

A total of 357 clinical Streptococcus pyogenes isolates collected between 1994 and 1999 in Rio de Janeiro city were tested for susceptibility to 10 antimicrobial drugs by agar-diffusion tests. All isolates were susceptible to penicillin, cephems, and vancomycin. High resistance rates were observed for tetracycline (43.1%) and trimethoprim/sulfamethoxazole (77.9%). Three isolates (0.8%) were resistant to erythromycin, and three exhibited intermediate susceptibility. Determination of the erythromycin MICs by the agar dilution method, showed 1.6% of erythromycin resistant isolates (the three erythromycin-resistant and the three erythromycin-intermediate isolates found by agar-diffusion test). Of the erythromycin-resistant isolates subjected to the double-disc diffusion test for erythromycin and clindamycin, three isolates expressed the iMLSB and three the M phenotype. The resistance phenotypes were confirmed by comparing the clindamycin MICs determined under normal testing conditions and those determined after induction by pre-growth in 0.06 microg/ml of erythromycin. Three ermTR and three mefA-containing isolates were detected by PCR. In strains belonging to the iMLSB phenotype, two clones were identified by PFGE following restriction with SmaI. M phenotype isolates could not be restricted with SmaI. Our results indicate a low rate of erythromycin resistance among S. pyogenes isolated in Rio de Janeiro, Brazil, and pointed to the presence of both resistance mechanisms found in streptococci.

Genetic and phenotypic features of Streptococcus pyogenes strains isolated in Brazil that harbor new emm sequences.

In the present study, 37 group A Streptococcus (GAS) strains belonging to 13 new emm sequence types identified among GAS strains randomly isolated in Brazil were characterized by using phenotypic and genotypic methods. The new types were designated st204, st211, st213, st809, st833, st854, st2904, st2911, st2917, st2926, st3757, st3765, and st6735. All isolates were susceptible to the antimicrobial agents tested, except to tetracycline. They all carried the speB gene, and 94.6% produced detectable SpeB. Most strains belonging to a given emm type had similar or highly related pulsed-field gel electrophoresis profiles that were distinct from profiles of strains of another type. The other characteristics were variable from isolate to isolate, although some associations were consistently found within some emm types. Unlike the other isolates, all type st213 isolates were speA positive and produced SpeA. Strains belonging to st3765 were T6 and opacity factor (OF) negative. Individual isolates within OF-positive emm types were associated with unique sof gene sequence types, while OF-negative isolates were sof negative by PCR. This report provides information on new emm sequence types first detected in GAS isolates from a geographic area not extensively surveyed. Such data can contribute to a better understanding of the local and global dynamics of GAS populations and of the epidemiological aspects of GAS infections occurring in tropical regions.

Phenotypic and genotypic characterization of Pediococcus strains isolated from human clinical sources.

Seventy-two strains of pediococci isolated from human clinical sources were characterized by conventional physiological tests, chromogenic enzymatic tests, analysis of whole-cell protein profiles (WCPP) by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and analysis of chromosomal DNA restriction profiles by pulsed-field gel electrophoresis (PFGE). Conventional tests allowed identification of 67 isolates: 52 strains were identified as Pediococcus acidilactici, 15 strains were identified as Pediococcus pentosaceus, and 5 strains were not identified because of atypical reactions. Analysis of WCPP identified all isolates since each species had a unique WCPP. By the WCPP method, the atypical strains were identified as P. acidilactici (two strains) and P. pentosaceus (three strains). The chromogenic substrate test with o-nitrophenyl-beta-D-glucopyranoside differentiated all 54 strains of P. acidilactici (negative reactions) and 13 (72%) of 18 strains of P. pentosaceus (positive reactions). Isolates of both species were shown to be nonclonal as revealed by the genetic diversity when chromosomal DNA was analyzed by PFGE. Using WCPP as the definitive identification procedure, P. acidilactici (28 of 54 strains; 51.8%) was more likely than P. pentosaceus (4 of 18 strains; 22.3%) to be isolated from blood cultures.

Leuconostoc pseudomesenteroides as a cause of nosocomial urinary tract infections.

The phenotypic and genotypic characterization of five clinical isolates of Leuconostoc pseudomesenteroides associated with nosocomially acquired urinary tract infections is described. All the strains were susceptible to chloramphenicol, clindamycin, erythromycin, gentamicin, and tetracycline; all were resistant to nalidixic acid, norfloxacin, and vancomycin; and all were intermediately affected by ampicillin and penicillin. Analysis of chromosomal DNA by pulsed-field gel electrophoresis after treatment with SmaI indicated a clonal relationship of the isolates. The results provide evidence for the possibility of nosocomial transmission of this unusual opportunistic, vancomycin-resistant pathogen.