[Resistance to non-beta-lactam antibiotics in the clinical isolates of Streptococcus pneumoniae of children in Latin America. SIREVA II, 2000-2005]. / Resistencia a antibióticos no betalactámicos de aislamientos invasores de Streptococcus pneumoniae en niños latinoamericanos. SIREVA II, 2000-2005.

OBJECTIVE: To examine the development of resistance to erythromycin, chloramphenicol, trimethoprim-sulfamethoxazole (TMP-SMZ), and vancomycin of the invasive isolates of Streptococcus pneumoniae obtained from children in 10 Latin American/Caribbean countries during six years of surveillance. METHODS: Analysis of 8 993 isolates of S. pneumoniae recovered in 2000-2005 from children with invasive infections, who were less than 6 years of age, and from Argentina, Brazil, Chile, Colombia, Cuba, Dominican Republic, Mexico, Paraguay, Uruguay, or Venezuela. Antibiotic susceptibility was determined through the methods established and standardized by the SIREVA project. Multidrug resistance was defined as: resistance to three or more antibiotics of the same class; to the non-beta-lactams analyzed by this study; or, to the beta-lactams evaluated by a previous study, in which 37.8% of these isolates showed decreased susceptibility to penicillin. RESULTS: Some degree of resistance was found to TMP-SMZ and erythromycin (56.4% and 15.4% of the isolates studied, respectively), with 4.6% highly resistant to chloramphenicol. All isolates were susceptible to vancomycin. The highest prevalence of TMP-SMZ resistance was observed in the pneumonia isolates; and that of erythromycin, in cases of sepsis (61.6% and 25.5%, respectively; P < 0.01). The highest prevalence of TMP-SMZ resistance was found in Brazil (71.9%), and that of erythromycin, in Mexico (38.2%) and Venezuela (32.9%). The 14, 6B, 19F, and 23F serotypes were most often associated with resistance to the antibiotics in the study. CONCLUSIONS: High and increasing rates of isolates resistant to TMP-SMZ and erythromycin were observed, as well as a decreasing percentage of isolates resistant to chloramphenicol. These trends highlight differences among the countries studied.

Resistencia a antibióticos no betalactámicos de aislamientos invasores de Streptococcus pneumoniae en niños latinoamericanos: SIREVA II, 2000-2005 / Resistance to non-beta-lactam antibiotics in the clinical isolates of Streptococcus pneumoniae of children in Latin America: SIREVA II, 2000-2005

OBJETIVO. Determinar la evolución de la resistencia a la eritromicina, el cloranfenicol, el trimetoprim-sulfametozaxol (SXT) y la vancomicina de aislamientos invasores de Streptococcus pneumoniae obtenidos de niños de 10 países de América Latina y del Caribe en seis años de vigilancia. MÉTODOS. Se analizaron 8 993 aislamientos de S. pneumoniae recuperados entre 2000 y 2005 de niños menores de 6 años con infecciones invasoras, procedentes de Argentina, Brasil, Chile, Colombia, Cuba, México, Paraguay, República Dominicana, Uruguay y Venezuela. La sensibilidad a los antibióticos se determinó mediante los métodos establecidos y estandarizados en el proyecto SIREVA. La resistencia a múltiples antibióticos se definió como la resistencia a tres o más familias de antibióticos, de los no betalactámicos analizados en este estudio o de los betalactámicos evaluados en un estudio previo en el que 37,8% de estos aislamientos presentaron sensibilidad disminuida a la penicilina. RESULTADOS. Se encontró algún grado de resistencia al SXT y la eritromicina (56,4% y 15,4% de los aislamientos estudiados, respectivamente) y 4,6% presentó alta resistencia al cloranfenicol. Todos los aislamientos fueron sensibles a la vancomicina. Se observó la mayor frecuencia de resistencia al SXT en los aislamientos de neumonía y a la eritromicina en los casos de sepsis (61,6% y 25,5%, respectivamente; P < 0,01). La mayor frecuencia de resistencia al SXT se observó en Brasil (71,9%) y a la eritromicina en México (38,2%) y Venezuela (32,9%). Los serotipos 14, 6B, 19F y 23F fueron los que más frecuentemente se asociaron con la resistencia a los antibióticos estudiados. CONCLUSIONES. Se observó una elevada y creciente frecuencia de aislamientos resistentes al SXT y la eritromicina, y una disminución en la proporción de aislamientos resistentes al cloranfenicol. Estas tendencias mostraron diferencias entre los países estudiados.

OBJECTIVE. To examine the development of resistance to erythromycin, chloramphenicol, trimethoprim-sulfamethoxazole (TMP-SMZ), and vancomycin of the invasive isolates of Streptococcus pneumoniae obtained from children in 10 Latin American/Caribbean countries during six years of surveillance. METHODS. Analysis of 8 993 isolates of S. pneumoniae recovered in 2000­2005 from children with invasive infections, who were less than 6 years of age, and from Argentina, Brazil, Chile, Colombia, Cuba, Dominican Republic, Mexico, Paraguay, Uruguay, or Venezuela. Antibiotic susceptibility was determined through the methods established and standardized by the SIREVA project. Multidrug resistance was defined as: resistance to three or more antibiotics of the same class; to the non-beta-lactams analyzed by this study; or, to the beta-lactams evaluated by a previous study, in which 37.8% of these isolates showed decreased susceptibility to penicillin. RESULTS. Some degree of resistance was found to TMP-SMZ and erythromycin (56.4% and 15.4% of the isolates studied, respectively), with 4.6% highly resistant to chloramphenicol. All isolates were susceptible to vancomycin. The highest prevalence of TMP-SMZ resistance was observed in the pneumonia isolates; and that of erythromycin, in cases of sepsis (61.6% and 25.5%, respectively; P < 0.01). The highest prevalence of TMP-SMZ resistance was found in Brazil (71.9%), and that of erythromycin, in Mexico (38.2%) and Venezuela (32.9%). The 14, 6B, 19F, and 23F serotypes were most often associated with resistance to the antibiotics in the study. CONCLUSIONS. High and increasing rates of isolates resistant to TMP-SMZ and erythromycin were observed, as well as a decreasing percentage of isolates resistant to chloramphenicol. These trends highlight differences among the countries studied

Susceptibilidad antimicrobiana y factores de virulencia en especies de Enterococcus causantes de infecciones pediátricas en Cuba / Antimicrobial susceptibility and virulence factors found in Enterococcus species causing pediatric infections

Objetivos: conocer la prevalencia de especies de enterococos causantes de infecciones en niño, su virulencia, susceptibilidad antimicrobiana e infecciones más frecuentes. Métodos: se estudiaron 156 aislamientos de enterococos causantes de infecciones en niños ingresados en el Hospital Octavio de la Concepción y La Pedraja, Holguín. Se llevó a cabo la identificación de especies y detección de factores de virulencia por métodos fenotípicos. Se determinó la concentración inhibitoria mínima para vancomicina, ciprofloxacina, ampicilina, gentamicina, estreptomicina, cloranfenicol, nitrofurantoína, eritromicina, tetraciclina, levofloxacina y norfloxacina, según las normas del Clinical Laboratory Standard Institute. Resultados: las cepas se aislaron de sangre, orina, heridas quirúrgicas, piel y tejido blando, oído medio, vulva, punta de catéter, líquido peritoneal, vagina, esperma, líquido cefalorraquídeo, y otras fuentes. La especie prevalente fue Enterococcus faecalis, seguido por Enterococcus faecium, Enterococcus gallinarum y Enterococcus raffinossus, Enterococcus casseliflavus y Enterococcus avium. La producción de proteasa y hemolisina se detectó solo en E. faecalis. E. faecium mostró porcentajes más altos de resistencia a ampicilina, ciprofloxacina, norfloxacina, levofloxacina, mientras que E. faecalis mostró más resistencia a eritromicina, cloranfenicol y tetraciclina. De E. faecalis 58 aislamientos y 12 de E. faecium presentaron altos niveles de resistencia a gentamicina, así como 45 y 32,3 por ciento mostraron altos niveles de resistencia para estreptomicina, respectivamente. Se detectó una cepa de E. faecium resistente a vancomicina. Conclusiones: la circulación de cepas de enterococos virulentas y resistentes a diferentes antibióticos indica la necesidad del control de la infección enterocócica..........(AU)

Objectives: to find out the prevalence of Enterococcus species causing child infections, their virulence, antimicrobial susceptibility and most frequent infections. Methods: One hundred and fifty six isolations from Enterococcus causing infections in children, admitted to Octavio de la Concepción y La Pedraja hospital in Holguin, were studied. The phenotypical methods allowed identifying species and detecting virulence factors. Following Clinical Laboratory Standard Institute requirements, minimal inhibitory concentration was determined for vancomycin, ciprofloxacin, ampicilline, gentamycin, streptomycin, chloranphenicol, nitrofurantoine, erythromycin, tetracycline, levofloxacin and norfloxacine. Results: Strains were isolated from the blood, the urine, surgical wounds, the skin and the soft tissue, the middle ear, the vulva, catheter point, peritoneal fluid, vagina, sperm, cerebrospinal fluid, and from other sources. The prevailing species was Enterococcus faecalis followed by Enterococcus faecium, Enterococcus gallinarum y Enterococcus raffinossus, Enterococcus casseliflavus y Enterococcus avium. Protease and hemolysin was only detected in E. faecalis. E. faecium accounted for the highest percentage of resistance to ampicillin, ciprofloxacine, norfloxacin, levofloxacin whereas E. faecalis was more resistant to erythromycin, chloranphenicol and tetracycline. Fifty eight E. faecalis isolates and 12 E. faecium isolates exhibited the highest levels of resistance to gentamycin, as well as 45 percent and 32.3 percent of samples had high levels of resistance to streptomycin respectively. A vancomycin-resistent E. faecium strain was detected. Conclusions: The circulation of virulent and multiple resistant Enterococcus strains indicates that Enterococcus infection need to be controlled and the importance of monitoring antimicrobial susceptibility ....................(AU)

Susceptibilidad antimicrobiana y factores de virulencia en especies de Enterococcus causantes de infecciones pediátricas en Cuba / Antimicrobial susceptibility and virulence factors found in Enterococcus species causing pediatric infections

Objetivos: conocer la prevalencia de especies de enterococos causantes de infecciones en niño, su virulencia, susceptibilidad antimicrobiana e infecciones más frecuentes. Métodos: se estudiaron 156 aislamientos de enterococos causantes de infecciones en niños ingresados en el Hospital Octavio de la Concepción y La Pedraja, Holguín. Se llevó a cabo la identificación de especies y detección de factores de virulencia por métodos fenotípicos. Se determinó la concentración inhibitoria mínima para vancomicina, ciprofloxacina, ampicilina, gentamicina, estreptomicina, cloranfenicol, nitrofurantoína, eritromicina, tetraciclina, levofloxacina y norfloxacina, según las normas del Clinical Laboratory Standard Institute. Resultados: las cepas se aislaron de sangre, orina, heridas quirúrgicas, piel y tejido blando, oído medio, vulva, punta de catéter, líquido peritoneal, vagina, esperma, líquido cefalorraquídeo, y otras fuentes. La especie prevalente fue Enterococcus faecalis, seguido por Enterococcus faecium, Enterococcus gallinarum y Enterococcus raffinossus, Enterococcus casseliflavus y Enterococcus avium. La producción de proteasa y hemolisina se detectó solo en E. faecalis. E. faecium mostró porcentajes más altos de resistencia a ampicilina, ciprofloxacina, norfloxacina, levofloxacina, mientras que E. faecalis mostró más resistencia a eritromicina, cloranfenicol y tetraciclina. De E. faecalis 58 aislamientos y 12 de E. faecium presentaron altos niveles de resistencia a gentamicina, así como 45 y 32,3 por ciento mostraron altos niveles de resistencia para estreptomicina, respectivamente. Se detectó una cepa de E. faecium resistente a vancomicina. Conclusiones: la circulación de cepas de enterococos virulentas y resistentes a diferentes antibióticos indica la necesidad del control de la infección enterocócica...

Objectives: to find out the prevalence of Enterococcus species causing child infections, their virulence, antimicrobial susceptibility and most frequent infections. Methods: One hundred and fifty six isolations from Enterococcus causing infections in children, admitted to Octavio de la Concepción y La Pedraja hospital in Holguin, were studied. The phenotypical methods allowed identifying species and detecting virulence factors. Following Clinical Laboratory Standard Institute requirements, minimal inhibitory concentration was determined for vancomycin, ciprofloxacin, ampicilline, gentamycin, streptomycin, chloranphenicol, nitrofurantoine, erythromycin, tetracycline, levofloxacin and norfloxacine. Results: Strains were isolated from the blood, the urine, surgical wounds, the skin and the soft tissue, the middle ear, the vulva, catheter point, peritoneal fluid, vagina, sperm, cerebrospinal fluid, and from other sources. The prevailing species was Enterococcus faecalis followed by Enterococcus faecium, Enterococcus gallinarum y Enterococcus raffinossus, Enterococcus casseliflavus y Enterococcus avium. Protease and hemolysin was only detected in E. faecalis. E. faecium accounted for the highest percentage of resistance to ampicillin, ciprofloxacine, norfloxacin, levofloxacin whereas E. faecalis was more resistant to erythromycin, chloranphenicol and tetracycline. Fifty eight E. faecalis isolates and 12 E. faecium isolates exhibited the highest levels of resistance to gentamycin, as well as 45 percent and 32.3 percent of samples had high levels of resistance to streptomycin respectively. A vancomycin-resistent E. faecium strain was detected. Conclusions: The circulation of virulent and multiple resistant Enterococcus strains indicates that Enterococcus infection need to be controlled and the importance of monitoring antimicrobial susceptibility...

Enzyme-linked immunosorbent assay for quantitative determination of capsular polysaccharide production in Streptococcus pneumoniae clinical isolates.

A simple, specific, sensitive and reproducible ELISA has been developed to quantify the level of CPS (capsular polysaccharide) production in supernatants of Streptococcus pneumoniae cell cultures. CPSs from Strep. pneumoniae have been widely used as vaccine antigens. The quantification method is based on two type-23F serotype-specific polyclonal antibodies: IgG, purified from sera of mice immunized with a pneumococcal type-23F CPS conjugate, used in the coating step, and a serotype-specific rabbit serum as the second antibody. Solutions of purified type-23F CPS were used as standards. The relationship between A(492) and type-23F CPS concentration was linear over the range 1-310 ng/ml (r=0.989), with 1 ng/ml as the lower limit of sensitivity. The specificity of ELISA was assessed because purified type-19F CPS and cell-wall polysaccharide samples were not detected after their evaluation by the ELISA described in the present study. Repeatability and intermediate precision of the assay were good, the coefficients of variation being 3 and 10% respectively. This ELISA allowed selection of an appropriate vaccine strain, for a natural polysaccharide vaccine, among several 23F pneumococcal clinical isolates and constituted a valuable analytical tool for Strep. pneumoniae fermentation and CPS purification follow-up.

High levels of multiple antibiotic resistance among 938 Haemophilus influenzae type b meningitis isolates from Cuba (1990-2002).

OBJECTIVES: A national surveillance study to determine antimicrobial susceptibility in Haemophilus influenzae type b isolated from cerebrospinal fluid was carried out in Cuba from 1990 to 2002. METHODS: Susceptibility to ampicillin, co-amoxiclav, cefotaxime, ceftriaxone, co-trimoxazole, tetracycline, chloramphenicol and rifampicin was tested by the microdilution method according to the NCCLS guidelines. RESULTS: The 34 participating laboratories recovered 938 consecutive, non-identical isolates. All the isolates were retrieved from children aged <5 years. The mean number of isolates collected by year in the pre-vaccination era (1990-1998) was 93; after vaccination, 57 isolates were reported in 1999, 31 in 2000, four in 2001 and five in 2002. Resistance to ampicillin, co-trimoxazole, tetracycline and chloramphenicol was 46.3% (all beta-lactamase-positive), 51.3%, 33.2% and 44.0%, respectively. Ampicillin-resistant beta-lactamase-negative strains were not detected. All strains were susceptible to co-amoxiclav, cefotaxime, ceftriaxone and rifampicin. Ampicillin resistance was strongly associated with resistance to tetracycline, co-trimoxazole and chloramphenicol (P<0.001). Multidrug resistance was present in 43.8% of isolates. The most prevalent phenotype was resistance to ampicillin/chloramphenicol/tetracycline/co-trimoxazole, which was detected in 29.2% of strains overall. An increase in the prevalence of resistance to these antibiotics was observed from 1990 to 2000 in the range 40.7%-54.8% for ampicillin, 40.1%-51.6% for chloramphenicol, 45.4%-58.1% for co-trimoxazole and 23%-45.2% for tetracycline. CONCLUSIONS: In Cuba, the widespread vaccination against Haemophilus influenzae type b prevented a large number of meningitis cases in children caused by strains resistant to multiple antibiotics.

Resultados y experiencias de la vigilancia nacionalde meningitis bacteriana en Cuba / Resultados y experiencias de la vigilancia nacional

Objetivos: Desde 1998 se implementó en Cuba un nuevo sistema de vigilancia nacional de meningitis bacteriana (VNMB) con el objetivo de lograr información en correspondencia con el desarrollo científico actual y las exigencias del programa nacional de prevención y control de estas infecciones. Resultados: Este sistema permitió medir el impacto de la vacuna contra H. influenzae tipo b (Hib) en 1999 sin necesidad de un proyecto y gastos adicionales, demostrando la reducción inmediata de la incidencia (52por ciento). Hasta el 2002 fueron reportados 82 niños < 5 años de los cuales 61(74,4por ciento) no estaban vacunados. Entre los vacunados hubo 14 (66,7por ciento) con una sola dosis aplicada.A partir de esta intervención S. pneumoniae resultó el principal agente causal de meningitis. Los serogrupos/ serotipos más frecuentes han sido 19, 6, 14, 18, 1, 7 y 23, mostrando un 40por ciento de susceptibilidad disminuida a la penicilina. Por primera vez identificamos: - fuerte asociación (RR > 20) de los estudiantes internos de primaria con la enfermedad, - incidencia elevada entre amas de casa (> 2/100 000) y jubilados (> 8/100 000), - fuerte asociación (Razón de disparidad > 20) de la muerte con jubilados y amas de casa en el análisis multivariado. Los resultados microbiológicos posibilitaron la caracterización de agentes y la política antibiótica más adecuada. La letalidad general de la meningitis neumocócica fue > 32 por ciento y fundamentalmente en ancianos. Conclusiones: LaVNMB aportó novedosa información clínico - epidemiológica y microbiológica, permitiendo ampliar el marco integral de los conocimientos y garantizando acciones más efectivas y eficientes en un proyecto multidisciplinario e intersectorial generalizado (AU)