Sickle cell disease children's gut colonization by extended-spectrum ß-lactamase (ESBL)-producing Enterobacterales: an antibiotic prophylaxis effect?

Introduction. Sickle cell disease (SCD) children have a high susceptibility to pneumococcal infection. For this reason, they are routinely immunized with pneumococcal vaccines and use antibiotic prophylaxis (AP).Hypothesis/Gap Statement. Yet, little is known about SCD children's gut microbiota. If antibiotic-resistant Enterobacterales may colonize people on AP, we hypothesized that SCD children on AP are colonized by resistant enterobacteria species.Objective. To evaluate the effect of continuous AP on Enterobacterales gut colonization from children with SCD.Methodology. We analysed 30 faecal swabs from SCD children on AP and 21 swabs from children without the same condition. Enterobacterales was isolated on MacConkey agar plates and identified by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) (bioMérieux, Marcy l'Etoile, France). We performed the antibiogram by Vitek 2 system (bioMérieux, Marcy l'Etoile, France), and the resistance genes were identified by multiplex PCR.Results. We found four different species with resistance to one or more different antibiotic types in the AP-SCD children's group: Escherichia coli, Klebsiella pneumoniae, Citrobacter freundii, and Citrobacter farmeri. Colonization by resistant E. coli was associated with AP (prevalence ratio 2.69, 95 % confidence interval [CI], 1.98-3.67, P<0.001). Strains producing extended-spectrum ß-lactamases (ESBL) were identified only in SCD children, E. coli, 4/30 (13 %), and K. pneumoniae, 2/30 (7 %). The ESBL-producing Enterobacterales were associated with penicillin G benzathine use (95 % CI, 22.91-86.71, P<0.001). CTX-M-1 was the most prevalent among ESBL-producers (3/6, 50 %), followed by CTX-M-9 (2/6, 33 %), and CTX-M-2 (1/6, 17 %).Conclusion. Resistant enterobacteria colonize SCD children on AP, and this therapy raises the chance of ESBL-producing Enterobacterales colonization. Future studies should focus on prophylactic vaccines as exclusive therapy against pneumococcal infections.

Antimicrobial-resistant enterobacteria in surface waters with fecal contamination from urban and rural communities.

INTRODUCTION: Inadequate wastewater treatment and fecal contamination have a strong environmental impact on antimicrobial resistance (AMR). This study evaluated the profile of AMR enterobacteria and fecal contamination from four surface waters: Jiquiriça-Brejões River and Cabrito, Tororó, and Abaeté Lagoons. METHODS: We analyzed AMR ß-lactamase genes using the polymerase chain reaction method and fecal contamination using Coliscan®. RESULTS: We found high levels of fecal contamination, ß-lactamase producers, and AMR genes (blaOXA-48, blaSPM, and blaVIM) in all waterbodies. CONCLUSIONS: Poor sanitation evidenced by fecal contamination and human activities around these surface waters contributed to the distribution and increase in AMR enterobacteria.

Antimicrobial-resistant enterobacteria in surface waters with fecal contamination from urban and rural communities

Abstract INTRODUCTION: Inadequate wastewater treatment and fecal contamination have a strong environmental impact on antimicrobial resistance (AMR). This study evaluated the profile of AMR enterobacteria and fecal contamination from four surface waters: Jiquiriça-Brejões River and Cabrito, Tororó, and Abaeté Lagoons. METHODS: We analyzed AMR β-lactamase genes using the polymerase chain reaction method and fecal contamination using Coliscan®. RESULTS: We found high levels of fecal contamination, β-lactamase producers, and AMR genes (blaOXA-48, blaSPM, and blaVIM) in all waterbodies. CONCLUSIONS: Poor sanitation evidenced by fecal contamination and human activities around these surface waters contributed to the distribution and increase in AMR enterobacteria.

Dissemination of the ST-103 clonal complex serogroup C meningococci in Salvador, Brazil.

Invasive meningococcal disease (IMD) is a major public health problem worldwide. An epidemic of serogroup C (NmC) IMD occurred in 2010 in the city of Salvador. In this study, we describe the antigenic and genetic characterization of meningococcal isolates collected from meningitis cases in Salvador from 2001 to 2012. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were performed for the analysis of IMD isolates. A total of 733 cases were identified, and the serogroup was determined for 391 (53.0%) of these. Most cases were caused by NmC (53%) or B (47%). The most prevalent strains were B:4,7:P1.19,15 (32.9%; 129/391) and C:23:P1.14-6 (28.6%; 112/391). Based on PFGE/MLST analysis, 71.3% (77/108 PFGE-tested isolates) clustered as two clones of sequence type ST-3779 and ST-3780, both belonging to the ST-103 clonal complex. ST-3779 has been detected in Salvador since 1996 and together with ST-3780 became predominant after 2005. There was a predominance of C:23:P1.14-6, ST-3779/3780 in Salvador during the period of 2007-2012, establishing a major clonal lineage, which remained in the community for a long time; this has serious implications for public health, particularly in terms of prevention and control strategies of IMD.

Temporal trends and clonal diversity of penicillin non-susceptible pneumococci from meningitis cases from 1996 to 2012, in Salvador, Brazil.

BACKGROUND: Hospital-based surveillance for pneumococcal meningitis has been conducted since January 1996 in the city of Salvador, Brazil. The purpose of this study was to describe the temporal evolution of Penicillin Non-Susceptible Streptococcus pneumoniae (PNSSP) in regards to serotype distributions and clonal diversity recovered from meningitis cases over 17 years. METHODS: Broth microdilution was used to identify pneumococcal isolates that were PNSSP (Minimum Inhibitory Concentration > 0.12 µg/ml). The annual incidence rate of meningitis cases was calculated. Serotyping was defined using multiplex polymerase chain reaction assays and quellung reaction. Genetic diversity of PNSSP isolates was assessed using both pulsed-field gel electrophoresis (PFGE) and Multilocus Sequence Typing (MLST) analyses. RESULTS: A total of 854 cerebrospinal fluid (CSF) culture pneumococcal isolates were tested by broth microdilution method and serotyped. A total of 173 (20.3%) were penicillin non-susceptible (PNSSP) (Minimum Inhibitory concentration ≥ 0.12 µg/ml). The annual incidence of meningitis cases declined from 1.65/100,000 population (1996) to 0.2/100,000 population in 2012 and the rate due to PNSSP declined 82% over the 17-years of surveillance. PNSSP isolates were restricted to 13 serotypes, being the most common ones serotypes 14 (45.1%; 78/173), 23 F (19.1%; 33/173), 6B (14.4%; 25/173), 19 F (9.2%; 16/173) and 19A (5.2%; 9/173). Among the PNSSP isolates, 94% had serotypes represented in the 10-valent conjugate vaccine (PCV10). The predominant serotype 14 clonal groups were identified as PFGE group A/multilocus sequence type 66 (ST66) [35.3% (61/173)] and PFGE group GK/ST156 [4.6% (8/173)], the latter one associated with high level resistance to penicillin and ceftriaxone. CONCLUSIONS: Our results show sustained reductions in pneumococcal meningitis cases in the Metropolitan region of Salvador from 1996 to 2012. This might reflect a beneficial impact of conjugate vaccines. Continued surveillance and further studies need to be conducted to better understanding on PCV10 vaccine impact.

Transmission of Streptococcus pneumoniae in an urban slum community.

BACKGROUND: Inhabitants of slum settlements represent a significant proportion of the population at risk for pneumococcal disease in developing countries. METHODS: We conducted a household survey of pneumococcal carriage among residents of a slum community in the city of Salvador, Brazil. RESULTS: Among 262 subjects, 95 (36%) were colonized with Streptococcus pneumoniae. Children <5 years of age (OR, 8.0; 95% CI, 3.5-18.6) and those who attended schools (OR, 2.7, 95% CI, 1.2-6.0) had significantly higher risk of being colonized. Of 94 isolates obtained from colonized individuals, 51% had serotypes included in the seven-valent pneumococcal conjugate vaccine. Overall, 10% (9 of 94 isolates) were nonsusceptible to penicillin and 28% (27 of 94 isolates) were resistant to cotrimoxazole. BOX-PCR, PFGE and MLST analyses found that 44% of the carriage isolates belonged to 14 distinct clonal groups. Strains of the same clonal group were isolated from multiple members of 9 out of the 39 study households. Nineteen carriage isolates had genotypes that were the same as those identified among 362 strains obtained from active surveillance for meningitis. CONCLUSIONS: The study's findings indicate that there is significant intra- and inter-household spread of S. pneumoniae in the slum community setting. However, a limited number of clones encountered during carriage among slum residents were found to cause invasive disease.

Epidemiologia molecular da meningite meningocócica em Salvador-BA / Molecular epidemiology of meningococcal meningitis in Salvador-BA

Atualmente a Neisseria Meningitidis é a principal causa de Meningite Bacteriana em todo mundo. Este estudo teve como objetivo consolidar os dados clínicos, epidemiológicos e microbiológicos da Meningite Meningocócica em Salvador, Brasil, num período de onze anos, decorrente de um sistema de vigilância hospitalar. Entre Fevereiro de 1996 e Dezembro de 2006, foram identificados os casos de Meningite Meningocócica comprovados por cultura positiva no Hospital Couto Maia, Salvador, BA. Dados demográficos e clínicos foram obtidos através de entrevista e revisão de prontuário. Anti-soro e anticorpos monoclonais foram usados para determinar os sorogrupos e sorotipos: sorosubtipos, respectivamente. Durante este período, dois métodos simples para genotipagem da Neisseria Meningitidis foram utilizados, um baseado na amplificação de elementos repetitivos por PCR (NgRep-PCR) e um outro baseado em RFLP (Polimorfismo dos fragmentos de restrição) o MLRFT (Tipagem de fragmentos de restrição de multilocus). Foram identificados 648 casos de Meningite Meningocócica, com taxa de letalidade de 11,3 por cento. A incidência anual média entre 324 casos residentes em Salvador foi de 1,20 casos/100.000 habitantes. A incidência foi maior em crianças menores de 4 anos de idade (9,77 casos/100.000 habitantes). Dos 575 isolados com resultados de sorogrupo, 79,3 por cento, 18,1 por cento, 1,7 por cento e 0,9 foram classificados como B, C, W135 e Y, respectivamente. Um único sorotipo: sorosubtipo (4,7:P1.19,15) foi responsável por 62,7 por cento de todos os casos. As análises utilizando os métodos de NgRep-PCR e MLRFT demonstraram padrões genotípicos com alta, reprodutibilidade, eficiência e conveniência, permitindo identificar os clones hipervirulentos, tais como os do complexo ET-5 encontrados em nosso estudo. Estes métodos foram comparáveis com o padrão ouro (MLST- tipagem por sequenciamento de multilocus), demonstrando algumas vantagens: execução rápida, simples, de baixo custo e de fácil aplicação em laboratórios de países em desenvolvimento. Em conclusão, um serviço de vigilância continuado é necessário para caracterizarmos cepas e para definir estratégias futuras no controle e prevenção da meningite meningocócica.

Population-based survey of antimicrobial susceptibility and serotype distribution of Streptococcus pneumoniae from meningitis patients in Salvador, Brazil.

Penicillin-nonsusceptible strains were isolated from 15% of 303 individuals with pneumococcal meningitis identified during a 4-year surveillance study in Salvador, Brazil. The estimated rate of coverage of the seven-valent conjugate vaccine was 74% among patients <5 years of age and 94% among those infected with nonsusceptible isolates, indicating that the use of conjugate vaccines may be an approach to the control of emerging penicillin resistance in Brazil.

Caracterização fenotípica e genotípica de isolados de Neisseria meningitidis de casos de meningite do Hospital Couto Maia, em Salvador-Bahia / Phenotypic and genotypic characterization of isolates of Neisseria meningitidis from cases of meningitis at the Hospital Couto Maia, in Salvador, Bahia

Infecções por Neisseria meningitidis estão associadas a altos índices de morbi-mortalidade. Em Salvador, a incidência anual média da meningite meningocócica, no período de fevereiro de 1996 a janeiro de 1999 foi de 2,25/100000 hab., acometendo, principalmente, crianças menores de 02 anos (14,19/100. 000hab.), uma taxa de letal idade geral de 7,9% e 4,7% de seqüelas neurológicas. Assim, se faz necessário a disponibilidade de um método simples, eficaz na caracterização molecular deste patógeno, para um melhor entendimento na distribuição e transmissão da doença. Este trabalho, teve como objetivos, descrever a distribuição fenotípica e genotípica da Neisseria meningitidis isolada de casos de meningite de Salvador-BA e validar a utilização do método da NgREP-PCR em estudos de vigilância epidemiológica. A população do estudo foi formada por pacientes do Hospital Couto Maia, Salvador-BA, que tiveram cultura de líquor positiva para Neisseria meningitidis. Para a caracterização fenotípica, as amostras foram submetidas a testes de aglutinação, na determinação dos sorogrupos (A, B, C, W135 e Y) e testes de dot-blot, utilizando anticorpos monoclonais, na determinação dos 18 sorotipos e 18 sorosubtipos e a caracterização genotípica foi realizada pela Reação da Polimerase em Cadeia - NgREP-PCR. Durante o período do estudo foram identificados no HCM, 272 casos de meningite meningocócica, dos quais, 258 tiveram isolados analisados. A distribuição dos sorogrupos, identificou o predomínio de sorogrupo B, em 79,8% (206 de 258) dos isolados, seguido do sorogrupo C, em 18,6% (48 de 258), do sorogrupo W135, em 1,2% (3 de 258) e do sorogrupo Y em 0,4% (1 de 258), nenhum caso sorogrupo A foi identificado. A distribuição dos sorotipos e sorosubtipos demonstrou a predomínio de 4,7;P1.19, 15(63,2%); 4,7;P1.7,1 (4,3%); 2a:P1.2 (3,5%); 4,3:P1.3 (1,9%); Outros (24,8%). Foram caracterizados pelo NgREP-PCR 84,5% (218 de 258). Através da análise visual, os isolados foram...