Effect on Antimicrobial Resistance of a Policy Restricting Over-the-Counter Antimicrobial Sales in a Large Metropolitan Area, São Paulo, Brazil.

Although restricting over-the-counter (OTC) antimicrobial drug sales is recommended globally, no data track its effect on antimicrobial resistance (AMR) in bacteria. We evaluated the effect of a national policy restricting OTC antimicrobial sales, put in place in November 2010, on AMR in a metropolitan region of São Paulo, Brazil. We reviewed associations between antimicrobial sales from private pharmacies and AMR in 404,558 Escherichia coli and 5,797 Streptococcus pneumoniae isolates using a dynamic regression model based on a Bayesian approach. After policy implementation, a substantial drop in AMR in both bacterial species followed decreased amoxicillin and trimethoprim/sulfamethoxazole sales. Conversely, increased ciprofloxacin sales were associated with increased ciprofloxacin resistance, and extended spectrum ß-lactamases-positive E. coli isolates and azithromycin sales increases after 2013 were associated with increased erythromycin resistance in S. pneumoniae isolates. These findings suggest that restricting OTC antimicrobial sales may influence patterns of AMR, but multifaceted approaches are needed to avoid unintended consequences.

Pneumococcal nasopharyngeal carriage in infants of mothers immunized with 23V non-conjugate pneumococcal polysaccharide vaccine.

To assess the effects of maternal immunization on pneumococcal colonization in infants, pregnant women were assigned into three groups. The group Pregn Vac received the Pn23V during pregnancy, the group Puerp Vac received vaccine during immediate puerperium and the group No Vac received no vaccine. Nasopharyngeal samples were collected at 3 and 6 months. A total of 150 pregnant women were selected during the prenatal period. The proportion of pneumococcal carriage in at least one evaluation was in group Pregn Vac 22.2% (10/45), group Puerp Vac 23.4% (11/47) and group No Vac 21.2% (10/47), respectively. The most frequently isolated serotype in group Puerp Va and group No Vac was 6B and 6A. In the Pregn Vac, the most important serotype was 19F. Although this study was unable to demonstrate any effect of maternal vaccination in reducing pneumococcal colonization, reduction of colonization for serotype 6B in infants may be an important effect.

Analysis of invasive pneumonia-causing strains of Streptococcus pneumoniae: serotypes and antimicrobial susceptibility.

OBJECTIVES: To identify the most common pneumococcal serotypes in children hospitalized with invasive pneumonia, correlate isolated serotypes with those included in conjugate vaccines, and ascertain the sensitivity of the isolated pneumococcal strains to penicillin and other antibiotics. METHODS: From January 2003 to October 2008, a retrospective study of hospitalized children with a diagnosis of Streptococcus pneumoniae pneumonia was conducted at the university hospital of Universidade de São Paulo. Criteria for inclusion were: age greater than 29 days and less than 15 years, radiological and clinical diagnosis of pneumonia, and isolation of Streptococcus pneumoniae in blood cultures and/or pleural effusion. RESULTS: The study included 107 children. The most common serotypes were 14 (36.5%), 1 (16%), 5 (14.6%), 6B (6.3%) and 3 (4.2%). The proportion of identified serotypes contained in the heptavalent, 10-valent and 13-valent conjugate vaccines was 53.1, 86.5, and 96.9%, respectively. Pneumococcal strains were sensitive to penicillin (minimum inhibitory concentration, MIC ≤ 2 µg/mL) in 100 cases (93.5%) and displayed intermediate resistance (MIC = 4 µg/mL) in 7 cases (6.5%). No strains were penicillin-resistant (MIC ≥ 8 µg/mL) according to the Clinical and Laboratory Standards Institute 2008 standards. Tested isolates were highly sensitive to vancomycin, rifampicin, ceftriaxone, clindamycin, erythromycin, and chloramphenicol. CONCLUSIONS: Our results confirm a significant potential impact of conjugate vaccines, mainly 10-valent and 13-valent, on invasive pneumonia. Furthermore, susceptibility testing results show that penicillin is still the treatment of choice for invasive pneumonia in our setting.

Análise das cepas de Streptococcus pneumoniae causadores de pneumonia invasiva: sorotipos e sensibilidade aos antimicrobianos / Analysis of invasive pneumonia-causing strains of Streptococcus pneumoniae: serotypes and antimicrobial susceptibility

OBJETIVOS: Identificar os sorotipos de pneumococo mais frequentemente isolados de crianças internadas com pneumonia invasiva, comparar os sorotipos com os incluídos em vacinas conjugadas e analisar sua sensibilidade aos antimicrobianos mais utilizados na faixa etária pediátrica. MÉTODOS: Estudo descritivo, retrospectivo das pneumonias pneumocócicas identificadas em crianças internadas no hospital universitário da Universidade de São Paulo, no período de janeiro de 2003 a outubro de 2008. Os critérios de inclusão foram: faixa etária de 29 dias até 15 anos incompletos com diagnóstico clínico e radiológico de pneumonia e com cultura de sangue e/ou líquido pleural com crescimento de Streptococcus pneumoniae. RESULTADOS: Foram incluídas no estudo 107 crianças. Os sorotipos mais frequentes foram: 14 (36,5 por cento), 1 (16,7 por cento), 5 (14,6 por cento), 6B (6,3 por cento) e 3 (4,2 por cento). A proporção de sorotipos contidos na vacina conjugada heptavalente seria de 53,1 por cento, na vacina 10-valente de 86,5 por cento e na 13-valente seria de 96,9 por cento. De acordo com os padrões do Clinical and Laboratory Standards Institute 2008, 100 cepas (93,5 por cento) de pneumococos foram sensíveis à penicilina (concentração inibitória mínima, CIM < 2 µg/mL), 7 cepas (6,5 por cento) com resistência intermediária (CIM = 4 µg/mL) e nenhuma com resistência (CIM > 8 µg/mL). Verificamos alta taxa de sensibilidade para as cepas testadas para vancomicina, rifampicina, ceftriaxone, clindamicina, cloranfenicol e eritromicina. CONCLUSÕES: Nossos resultados confirmam um expressivo impacto potencial das vacinas conjugadas, principalmente pela 10-valente e 13-valente, sobre os casos de pneumonias invasivas. Os resultados de sensibilidade à penicilina evidenciam que a opção terapêutica de escolha para o tratamento das pneumonias invasivas continua sendo a penicilina.

OBJECTIVES: To identify the most common pneumococcal serotypes in children hospitalized with invasive pneumonia, correlate isolated serotypes with those included in conjugate vaccines, and ascertain the sensitivity of the isolated pneumococcal strains to penicillin and other antibiotics. METHODS: From January 2003 to October 2008, a retrospective study of hospitalized children with a diagnosis of Streptococcus pneumoniae pneumonia was conducted at the university hospital of Universidade de São Paulo. Criteria for inclusion were: age greater than 29 days and less than 15 years, radiological and clinical diagnosis of pneumonia, and isolation of Streptococcus pneumoniae in blood cultures and/or pleural effusion. RESULTS: The study included 107 children. The most common serotypes were 14 (36.5 percent), 1 (16 percent), 5 (14.6 percent), 6B (6.3 percent) and 3 (4.2 percent). The proportion of identified serotypes contained in the heptavalent, 10-valent and 13-valent conjugate vaccines was 53.1, 86.5, and 96.9 percent, respectively. Pneumococcal strains were sensitive to penicillin (minimum inhibitory concentration, MIC < 2 µg/mL) in 100 cases (93.5 percent) and displayed intermediate resistance (MIC = 4 µg/mL) in 7 cases (6.5 percent). No strains were penicillin-resistant (MIC > 8 µg/mL) according to the Clinical and Laboratory Standards Institute 2008 standards. Tested isolates were highly sensitive to vancomycin, rifampicin, ceftriaxone, clindamycin, erythromycin, and chloramphenicol. CONCLUSIONS: Our results confirm a significant potential impact of conjugate vaccines, mainly 10-valent and 13-valent, on invasive pneumonia. Furthermore, susceptibility testing results show that penicillin is still the treatment of choice for invasive pneumonia in our setting.

Laboratory-based surveillance of Streptococcus pneumoniae invasive disease in children in 10 Latin American countries: a SIREVA II project, 2000-2005.

BACKGROUND: For the last 14 years the Pan American Health Organization has been promoting surveillance of invasive pneumococcal disease in Latin American children for better understanding of the disease tendencies regarding capsular types circulation in each country and susceptibility to antimicrobials. METHODS: Laboratory-based surveillance data from 10 Latin American countries collected from 2000 to 2005 were analyzed, including serotype distribution and susceptibility to beta-lactam antibiotics. RESULTS: Although 61 different capsular types were identified during the 6-year surveillance, 13 serotypes accounted for 86% of all isolates. These were consistently the most prevalent throughout the study period with serotype 14 predominating. Diminished susceptibility to penicillin was detected in 38% of all Streptococcus pneumoniae isolates, with the highest prevalence in Dominican Republic and Mexico. Decreased susceptibility to penicillin increased in Brazil and Colombia whereas decreased high resistance rates was recorded in Chile. CONCLUSIONS: These data indicate that 10 countries of the Region continue to have high quality laboratory-based surveillance for pneumococcal disease thus generating valuable information so that healthcare decision makers may prioritize interventions. The heptavalent vaccine will potentially cover from 52.4% to 76.5% of strains causing invasive pneumococcal disease and the 13 valent from 76.7% to 88.3%.

Serotypes and clonal types of penicillin-susceptible streptococcus pneumoniae causing invasive disease in children in five Latin American countries.

We used multilocus sequencing typing (MLST) to determine the genetic backgrounds of 185 recent penicillin susceptible Streptococcus pneumoniae isolates with serotypes that most frequently cause invasive disease in preschool age children in five Latin American countries-Argentina, Brazil, Colombia, Mexico, and Uruguay. Most of the isolates were associated with pneumonia (90/185), meningitis (74/185), and bacteremia (17/185). The collection of strains included seven serotypes-14, 6B, 5, 1, 23 F-which represent the serotypes of S. pneumoniae most frequently associated with sterile site infections in children. Also included were strains expressing serotypes 7F and 3. Comparison of serotype and multilocus sequence type allowed division of the isolates into two groups: strains expressing serotypes 1, 5, 3, and 7 were represented by a relatively few sequence types while strains expressing serotypes 6B, 14, and 23 F showed great genetic diversity. The genetic diversity of serotypes 14, 6B, and 23 F may be related to the capacity of these serotypes to colonize the nasopharynx of healthy carriers during which opportunities for diversification through genetic exchanges can occur. The findings present an interesting contrast with the results of an earlier study in which over 80% of invasive penicillin- resistant serotype 14 and 23 isolates from the same countries were found to belong to as few as two pandemic clones of S. pneumoniae.

Occurrence of insertion sequences within the genomes and Tn1546-like elements of glycopeptide-resistant enterococci isolated in Brazil, and identification of a novel element, ISEfa5.

Insertion sequences (IS) occur widely within the Tn1546-like elements responsible for VanA glycopeptide resistance in enterococci from several countries. As such insertions can be used as epidemiological markers and for studying horizontal transfer of gene clusters, we investigated the distribution of IS6770, IS1542, IS1216V, IS1476, and IS1251 elements in 26 VanA Enterococcus faecium and 21 VanA Enterococcus faecalis from Brazil. PCR, using genomic DNA as a template, indicated that most of the isolates contained IS6770 (97%), IS1216V (87%) and IS1476 (72%) elements. IS1251 was also detected, but at a higher frequency in E. faecium (80%) than in E. faecalis (14%). None of the isolates harboured IS1542. Only two of 47 isolates had IS elements within their Tn1546-like elements; one possessed IS1251 between vanS and vanH, as reported in the United States; another possessed a novel IS element, designated ISEfa5, located between vanX and van Y. This novel element was found in the genomic DNA of 25 (96%) E. faecium and II (52%) E. faecalis. In stability studies, no IS-mediated changes were detected in the Tn1546-like elements of 25 vancomycin-resistant enterococci (VRE) monitored over 11 months. These results suggest that the occurrence of IS in Brazilian isolates is similar to that reported in American isolates, but that these elements occur rarely within the vanA gene clusters. As patterns of IS carriage did not correlate with the PFGE type of the VRE, the prevalence of IS elements in genomic DNA of VRE is not a useful epidemiological marker. However, the presence of IS-modified Tn1546-like elements, which appear to be rare in Brazil, could be a useful molecular marker in local epidemiological studies to monitor the evolution and horizontal transmission of VanA elements.

[Prevalence of serotypes and antimicrobial resistance of invasive strains of Streptococcus pneumoniae]. / Prevalência de sorotipos e resistência antimicrobiana de cepas invasivas do Streptococcus pneumoniae.

OBJECTIVE: To determine the prevalence of serotypes and antimicrobial susceptibility of invasive strains of Streptococcus pneumoniae and to discuss the implications of these findings for vaccine formulation. METHODS: Strains of Streptococcus pneumoniae obtained from normally sterile fluids from patients admitted with invasive diseases were isolated and identified at the Hospital de Clínicas, Universidade Federal de Uberlândia, state of Minas Gerais, and forwarded to Instituto Adolfo Lutz, state of São Paulo, for further identification, serotyping and determination of antimicrobial susceptibility. RESULTS: From April 1999 to March 2003, 148 invasive strains of Streptococcus pneumoniae were obtained. The age of patients ranged from 1 day to 88.83 years (mean: 21.33+/-25.82 years; median: 4.42 years). Eighty-four (56.7%) patients were male. The most common diagnoses were pneumonia (91 cases; 61.4%), meningitis (32 cases; 21.6%) and occult bacteremia (15 cases; 10.1%). Strains were isolated mostly from blood (76 occasions; 51.3%), pleural fluid (39 occasions; 26.3%) and cerebrospinal fluid (30 occasions; 20.2%). There were 23 different serotypes, and the most common were 14, 3, 1, 5, 6A, 6B and 18C. Among 30 (20.2%) oxacillin-resistant strains, 23 (15.5%) were confirmed as resistant to penicillin (12.8% intermediate resistance and 2.7% full resistance). Oxacillin-resistant strains were restricted to serotypes 14, 23F, 19A and 6B. Resistance to penicillin varied with age, being more common in children under two years of age (p = 0.0008). We observed decreased sensibility to sulfamethoxazole-trimethoprim (92 isolates; 63.4%), to erythromycin (12 isolates; 8.3%), to clindamycin (12 isolates; 8.7%), to ofloxacin (one strain; 0.8%) and to cefotaxime (three strains; 2%; also resistant to penicillin). All isolates were susceptible to chloramphenicol, rifampin and vancomycin. CONCLUSIONS: The decreased susceptibility to penicillin, detected in 15.5% of the strains was predominant in children under two years of age. There were 23 different Streptococcus pneumoniae serotypes. The 23-valent polysaccharide vaccine covers 82.6% of the serotypes and 90.2% of the invasive strains isolated in this population. In addition, 46.7% of the serotypes and 63.6% of the strains isolated from children until five years of age are covered in the currently available 7-valent conjugated vaccine (PN CRM7).

[Invasive pneumococcal strains isolated from children and adolescents in Salvador]. / Cepas invasivas de pneumococo isoladas de crianças e adolescentes em Salvador.

OBJECTIVES: To describe the antimicrobial resistance and serotype distribution of pneumococcal strains. METHODS: In a 57-month period, a laboratory-based surveillance of invasive pneumococcal strains from patients aged < 20 years was conducted. Pneumococcus was identified by means of tests for solubility in bile and optochin. Pneumococcal resistance to penicillin was screened by 1 micro g oxacillin disc and minimal inhibitory concentration was determined for the strains not susceptible to penicillin. Disc diffusion and broth microdilution methods were used for surveillance of resistance to other antimicrobials. Pneumococci were serotyped by means of the Neufeld-Quellung reactions. RESULTS: Of 70 patients, 57.1% were males. The mean age was 1.92 yrs (mean 3.19 +/- 3.66 yrs, range 1 month to 19.5 yrs); 52.9% and 81.4% were < 2 yrs and < 5 yrs, respectively. The strains were isolated from blood (91.4%), CSF (2.9%), pleural (2.9%), peritoneal (1.4%) and abscess (1.4%) fluids from patients with pneumonia (77.1%), fever without localizing signs (10.0%), meningitis (4.3%), others (8.6%). Resistance was detected to penicillin (20.0%), trimethoprim-sulfamethoxazole (65.7%), tetracycline (21.4%), ofloxacin (6.3%), erythromycin (5.7%), clindamycin (2.9%). All tested strains were susceptible to chloramphenicol and vancomycin. Among penicillin-resistant strains, high resistance was detected in one, the same that showed intermediate resistance to cefotaxime. The most frequent serotypes were: 14 (22.9%), 5 and 6A (10.0% each), 6B and 19F (8.6% each), 9V, 18C and 23F (5.7% each). Resistance to penicillin was detected in serotypes 14 (71.4%), 6B and 19F (14.3% each). CONCLUSIONS: Of 70 strains, 67.2% were classified as serotypes included in the heptavalent conjugate pneumococcal vaccine, as were all penicillin-resistant strains.

Cepas invasivas de pneumococo isoladas de crianças e adolescentes en Salvador / Invasive pneumococcal strains isolated from children and adolescents in Salvador

Objetivos: descrever resistencia antimicrobiana e sorotipos de cepas de pneumococo. Metodos: durante 57 meses, foi conduzida uma vigilancia de cepas invasivas de pneumococo de pacientes com idade maior de 20 anos. O pneumococo foi identificado pelos testes de solubilidade da bile e optoquina. A resistência a penicilina foi avaliada com o disco de oxacilina (I╡g) e, para as cepas não suscetiveis, foi determinada a concentração inibitariamonima. Provas de difusão de disco e de microdiluiτπo em placa foram utilizadas para avaliação da resistência a outros antimicrobianos...

Typing and molecular characterization of Streptococcus pneumoniae with reduced susceptibility to cefotaxime isolated in Latin America.

The treatment of systemic infections, especially meningitis, caused by Streptococcus pneumoniae nonsusceptible to third-generation cephalosporins, is extremely difficult due to the paucity of therapeutic options. The main objective of this study was to characterize isolates of S. pneumoniae with reduced susceptibility to cefotaxime (MICs, > or = 1 microg/ml) by different typing methods and to evaluate whether clonal dissemination of this pathogen had occurred among Latin American medical centers. A total of 46 isolates collected from respiratory tract specimens, blood cultures, cerebrospinal fluid, eye, and other sources were analyzed. The isolates were collected from Latin American medical centers located in Argentina, Brazil, Chile, Colombia, Mexico, and Uruguay through two multicenter surveillance programs, in 1997 and 1998. Isolates were serotyped and molecular typed by pulsed-field gel electrophoresis (PFGE) and automated ribotyping. Antimicrobial susceptibilities were determined to 19 drugs by reference broth microdilution methods. Ten isolates (21.7%) had cefotaxime MICs > or = 2 microg/ml, whereas 36 (78.3%) had cefotaxime MIC results at 1 microg/ml. All isolates were susceptible to gatifloxacin, levofloxacin, and vancomycin. The isolates were distributed among five major serotypes (%): 23F (39.1%), 14 (32.6%), 19F (23.9%), 9V (2.2%), and 6B (2.2%). However, distinct molecular patterns were detected among isolates with a unique serotype. Six and four PFGE patterns were identified among isolates with serotype 23F and 19F, respectively. When PFGE and automated ribotyping analyses were combined, four clusters were identified. The largest cluster (10 isolates) was represented by isolates with ribotype 18-2, major PFGE pattern I, and serotype 14. ATCC 700671 (international clone Spain 9V-3) also showed ribotype 18-2. This clone was detected in four countries: Argentina, Brazil, Chile, and Uruguay. A second cluster (8 isolates) were characterized by isolates with ribotype 17-4, PFGE type D, and serotype 23F, similar to ATCC 700669 (international clone Spain23F-1). Isolates from this cluster were identified in three countries: Brazil, Chile, and Mexico. Our results indicated that clonal dissemination of S. pneumoniae with reduced susceptibility to cefotaxime has occurred in Latin America mainly among serogroups 14, 19F, and 23F.