Susceptibility to carbapenems in 238 isolates of Pseudomonas spp. recovered from environmental samples was investigated. In 198 (128 isolates with intermediate sensitivity / resistance to imipenem (IMP) and / or meropenem (MER) and 70 susceptible to both antimicrobials) of the 238 isolates studied, the production of carbapenemases was investigated phenotypically and genotypically. 8.8% of the isolates were resistant to IMP and 25.2% to MER. Nine strains recovered from water showed phenotypically the production of MBL. All of them were identified as P. otitidis. The genes blaIMP, blaSPM, blaVIM, blaKPC, or blaNDM were not detected in none of these isolates. The presence of resistant bacteria in the environment would require actions of control and environmental sanitation due to the impact that could represent on public health.

Se investigó la sensibilidad a carbapenemes en 238 aislamientos de Pseudomonas spp. recuperados de muestras ambientales. En 198 (128 aislamientos con sensibilidad intermedia/resistencia a imipenem (IMP) y/o meropenem (MER) y 70 sensibles a ambos antimicrobianos) de los 238 aislamientos estudiados, se investigó fenotípica y genotípicamente la producción de carbapenemasas. El 8,8% de los aislamientos fue resistente a IMP y el 25,2% a MER. Nueve cepas recuperadas de agua mostraron fenotípicamente la producción de MBL. Todas fueron identificadas como P. otitidis. En ninguno de estos aislamientos se detectó la presencia de los genes blaIMP, blaSPM, blaVIM, blaKPC, ni blaNDM. La presencia de bacterias resistentes en el ambiente, exigiría acciones de control y saneamiento ambiental dado el impacto que podría representar para la salud pública.

Streptococcus agalactiae (SGB) es la primera causa de infecciones severas invasivas en el recién nacido. Penicilina es la droga de elección en la profilaxis intraparto que se realiza en la embarazada colonizada con SGB, pero en aquéllas alérgicas a los ß-lactámicos , eritromicina (ERI) y clindamicina (CLI) son las drogas de elección. Conocer los fenotipos de resistencia a ERI y CLI es crucial a la hora de prevenir esta severa enfermedad en los neonatos nacidos de gestantes colonizadas alérgicas a los ß-lactámicos. Se seleccionaron al azar 5 cepas con algún fenotipo de resistencia y cinco cepas sin ellos, mantenidas a -70 ºC, obtenidas de hisopados vagino-rectales de gestantes, entre los años 2004 y 2011. Los biotipos se determinaron con sueros del Statens Serum Institut Copenhagen, los fenotipos con test D-test, los genes ermB, ermTR y mef A con PCR . De las 5 cepas con fenotipo de resistencia, dos cepas fueron constitutivas, una inducible y dos fenotipos M. En los biotipos V y III se detectó el mayor número de genes de resistencia. El gen de resistencia más comunmente detectado fue el ermB seguido de mef A. Alguno de los genes de resistencia investigados fueron detectados en tres cepas sin fenotipo de resistencia. El conocer la asociación de biotipos, fenotipos y genes de resistencia a ERI y CLI, circulantes en la región, datos aún no existentes, colabora en la salud individual y colectiva en profilaxis y tratamientos y en el diseño de estrategias vacunales, aún en marcha.

Introduction: Although Streptococcus agalactiae is commensal of the gastrointestinal and genitourinary tract, it is the main cause of invasive diseases and mortality in newborns. The infection can be acquired through the aspiration of infected amniotic fluid or during the passage through the birth canal. Objectives: to compare the effectiveness of various methods for the conservation of Streptococcus agalactiae strains that can be reproducible and accessible to laboratories of low and medium complexity and guarantee their phenotypic and genotypic stability through a preservation method called continuous subculture, for the maintenance of the culture in an adequate environment with fresh medium transfers at variable interval periods. Methods: 40 strains of Streptococcus agalactiae were randomly selected, which were subjected to verification of viability, purity and genotypic and phenotypic characterization before and after being subjected to conservation, using identical environments, reactive and methodologies in both circumstances. Different means of preservation of strains were tested, which allowed laboratories of low and medium complexity their transfer to specialized centers for the proper surveillance of the organism. The strains were kept for nine months with subcultures that showed original characteristics. Results: The most effective environments were ATS-TA and LD-4 % -SO-20 ºC because they guaranteed viability, purity and genotypic and phenotypic stability of the strains. It was shown that the use of this environment is an adequate alternative for the conservation of Streptococcus agalactiae in laboratories where lyophilization and dessication are not available and are low cost, fast and very easy to use in regular practice. Conclusions: The ATS-TA and LD-4 % -SO-20 ºC environments are a good alternative for short periods of preservation and transportation of Streptococcus agalactiae strains in laboratories of low and medium complexity.

Introducción: Si bien Streptococcus agalactiae es comensal del tracto gastrointestinal y genitourinario, es la principal causa de enfermedades invasivas y de mortalidad en niños recién nacidos. La infección puede adquirirse a través de la aspiración de líquido amniótico infectado o durante el paso por el canal de parto. Objetivos: comparar la utilidad de diversos métodos de conservación de cepas de Streptococcus agalactiae que resulten reproducibles, accesibles a laboratorios de baja y mediana complejidad, y asegurar su estabilidad fenotípica y genotípica mediante el método de preservación llamado subcultivo continuo, para el mantenimiento del cultivo en medio adecuado con transferencias a medio fresco a intervalos variables. Métodos: Se seleccionaron al azar 40 cepas de Streptococcus agalactiae, las que se sometieron a verificación de viabilidad, pureza y caracterización fenotípica y genotípica antes y después de ser sometidas a conservación, utilizando idénticos medios, reactivos y metodología en ambas circunstancias. Se probaron diferentes medios de preservación de las cepas, que permitieran a laboratorios de baja y mediana complejidad su traslado a centros especializados para la vigilancia adecuada del organismo. Las cepas se conservaron durante nueve meses con subcultivos que mostraron las características originales. Resultados: Los medios más efectivos fueron ATS-TA y LD 4 %-SO-20 ºC, ya que garantizaron viabilidad, pureza y estabilidad fenotípica y genotípica de las cepas. Se demostró que el uso de este medio es una alternativa adecuada para la conservación de Streptococcus agalactiae en laboratorios donde la liofilización y la desecación no están disponibles y que son de bajo costo, rápidos y muy fáciles de usar en la práctica habitual. Conclusiones: los medios ATS-TA y LD 4 %-SO-20 ºC constituyen una buena alternativa para cortos períodos de preservación y transporte de cepas de Streptococcus agalactiae en laboratorios de baja y mediana complejidad.

Group B Streptococcus (GBS) is the leading cause of neonatal infections. Our purpose was to characterize GBS colonization in pregnant women, current serotypes, resistance phenotypes and genes associated with virulence. In Misiones, Argentina, there are no previous data on this topic. Vaginal-rectal swabs from 3125 pregnant women were studied between 2004 and 2010. GBS strains were identified by conventional and serological methods (Phadebact Strep B Test, ETC International, Bactus AB, Sweden). Serotypes were detected using Strep-B Latex (Statens Serum Institut, Denmark). Resistance phenotypes were determined by the double-disk test. Genes were studied by PCR. Maternal colonization was 9.38%. Resistance to erythromycin was 11.6%, and the constitutive phenotype was the predominant one. Serotype Ia was the most frequent, whereas serotypes IV, VI, VII and VIII were not detected. The lmb, bca and hylB genes were detected in more than 79% of the strains. In this study, the colonization rate with GBS and the serotype distribution were compared with studies reported in other areas of the country. The high resistance to erythromycin in Misiones justifies performing antibiotic susceptibility testing. The serotype distribution, the genes encoding putative virulence factors, and the patterns of resistance phenotypes of GBS may vary in different areas. They thus need to be evaluated in each place to devise strategies for prevention.

The aim of this work was to study the distribution of a gene associated with Streptococcus agalactiae (Group B Streptococcus, GBS) virulence in colonizing isolates from pregnant women and invasive isolates from newborn infants. We also aimed to contribute with useful data for epidemiological studies and formulation of a regional vaccine. A total of 34 colonizing isolates and 7 invasive isolates was analyzed for detection of rib gene by specific PCR. The rib gene was present in 87.8 % of the isolates, without significant differences (p>0.05) between colonizing and invasive ones. It was concluded that in the population studied, the rib gene in GBS strains was distributed comparably in colonizing isolates from pregnant women and invasive isolates from newborn infants with severe disease.

El objetivo de este trabajo fue estudiar la distribución de un gen asociado a la virulencia de Streptococcus agalactiae (Streptococcus Grupo B, SGB) en aislamientos colonizantes recuperados de mujeres embarazadas y aislamientos invasivos recuperados de neonatos, y de contribuir con datos útiles para estudios epidemiológicos y para la formulación de una vacuna regional. Un total de 34 aislamientos colonizantes y 7 aislamientos invasivos fueron analizados para detectar el gen rib por PCR específica. Se detectó la presencia del gen rib en el 87,8 % de los aislamientos, sin diferencias significativas (p>0,05) entre colonizantes e invasivos. Concluimos que en la población estudiada, el gen rib en cepas de SGB se distribuye comparablemente en aislamientos colonizantes recuperados de mujeres embarazadas y en aislamientos invasivos recuperados de neonatos con enfermedad severa.

Streptococcus agalactiae or Group B Streptococcus (GBS) is one of the major causes of serious neonatal infection. Given the importance of their detection, the aim of this study was to identify the types of blood (human, bovine, equine, and ovine blood) that provide better yields for the isolation and identification of GBS. They could be used in laboratories of low and medium complexity. In 30 GBS strains of collection studied, the best yields (number, size, degree of hemolysis of the colonies and results of CAMP's test obtained with bovine and ovine blood. We suggest the use of ovine or bovine blood for the recovery and identification of GBS in laboratories of medium and low complexity, thus avoiding the risk of using human blood.

Streptococcus agalactiae o Streptococcus grupo B (SGB) es una de las causas principales de infección neonatal grave. Dada la importancia de su detección, el objetivo de este estudio fue identificar los tipos de sangre (humana, bovina, equina y ovina) que provean mejor rendimiento para el aislamiento e identificación de SGB y que puedan ser utilizados en laboratorios de baja y mediana complejidad. En 30 cepas de SGB de colección estudiadas, los mejores rendimientos (número, tamaño, grado de hemólisis de las colonias y resultados de la Prueba de CAMP) se obtuvieron con sangre bovina y ovina. Sugerimos, el uso de sangre ovina o bovina para la recuperación e identificación de SGB en laboratorios de mediana y baja complejidad, evitando así el riesgo que significa el uso de sangre humana.

Esse estudo objetivou determinar os padrões de sensibilidade a antibióticos e as taxas de colonização de Streptococcus do grupo B (GBS) em uma população de mulheres grávidas. Entre janeiro de 2004 e dezembro de 2006, foram obtidos swabs vaginais-retais de 1105 mulheres no Hospital Dr. Ramon Madariaga, em Posadas, Missiones, Argentina. A positividade para GBS nas mulheres grávidas foi 7,6%. Um total de 62 cepas de GBS foi selecionado ao acaso para testes in vitro de sensibilidade a penicilina G, ampicilina, tetraciclina, levofloxacina, gatifloxacina, ciprofloxacina, quinupristina-dalfopristina, linezolida, vancomicina, rifampicina, trimetoprim-sulfametoxazol, nitrofurantoína, gentamicina, clindamicina e eritromicina, e determinação dos fenótipos de resistência. Não foi encontrada resistência à penicilina, ampicilina, quinupristina-dalfopristina, linezolida e vancomicina. Entre as cepas, 96,8%, 98,3%, 46,8% e 29,0% foram sensíveis à rifampicina, nitrofurantoína, trimetoprim-sulfametoxazol e tetraciclina, respectivamente. Para as quinolonas, a ordem de sensibilidade foi: gatifloxacina (98,4%) > levofloxacina (93,8%) > ciprofloxacina (64,5%). A taxa de resistência à eritromicina (9,7%) foi superior a de outros relatos na Argentina. Nenhuma das cepas apresentou alto nível de resistência à gentamicina. Devido a 50% das cepas de GBS terem apresentado MIC para gentamicina igual ou inferior a 8 mg/ml, correspondente à concentração usada em um dos meios seletivos recomendados para GBS, sugeriu-se ao menos em nossa população, o emprego de ácido nalidíxico e colistina em meios seletivos para melhorar a sensibilidade da triagem de culturas para GBS em mulheres grávidas.

This study was performed to determine the susceptibility patterns and the colonization rate of Group B Streptococcus (GBS) in a population of pregnant women. From January 2004 to December 2006, vaginal-rectal swabs were obtained from 1105 women attending Dr. Ramón Madariaga Hospital, in Posadas, Misiones, Argentina. The carriage rate of GBS among pregnant women was 7.6%. A total of 62 GBS strains were randomly selected for in vitro susceptibility testing to penicillin G, ampicillin, tetracycline, levofloxacin, gatifloxacin, ciprofloxacin, quinupristin-dalfopristin, linezolid, vancomycin, rifampicin, trimethoprim-sulfametoxazol, nitrofurantoin, gentamicin, clindamycin and erythromycin, and determination of resistance phenotypes. No resistance to penicillin, ampicillin, quinupristin-dalfopristin, linezolid, and vancomycin was found. Of the isolates examined 96.8%, 98.3%, 46.8%, and 29.0% were susceptible to rifampicin, nitrofurantoin, trimethoprim-sulfametoxazol and tetracycline, respectively. Rank order of susceptibility for the quinolones was: gatifloxacin (98.4%) > levofloxacin (93.5%) > ciprofloxacin (64.5%). The rate of resistance to erythromycin (9.7%) was higher than that of other reports from Argentina. High-level resistance to gentamicin was not detected in any of the isolates. Based on our finding of 50% of GBS isolates with MIC to gentamicin equal o lower than 8 µg/ml, a concentration used in one of the selective media recommended for GBS isolation, we suggested, at least in our population, the use of nalidixic acid and colistin in selective media with the aim to improve the sensitivity of screening cultures for GBS carriage in women.

E. coli é o principal agente envolvido na produção de diarréias nos países em desenvolvimento. O objetivo deste estudo foi avaliar o tempo de aparição da primeira infecção assintomática pelas diferentes categorias de E. coli causadoras de diarréias em 44 crianças desde o nascimento e durante os primeiros 20 meses de vida. Em todas as crianças estudadas foram detectadas algumas categorias de E. coli causadoras de diarréia ao longo dos 20 meses do estudo. Durante o estudo foram obtidas 510 E. coli causadoras de diarréia (33,5%) a partir das 1524 amostras colectadas das 44 crianças (31,4% EAggEC, 28,8% EPEC, 27,1%DAEC e 12,7% ETEC). Não foram detectadas nem EHEC e nem EIEC. A idade média para a colonização pelas E. coli produtoras de diarréia foi de 7,5 meses. O período médio até o desmame foi de 12,8 meses e a idade média para a introdução de alimentação mista foi de 3,8 meses. Uma incidência significativamente baixa de diarréia e infecções assintomáticas foi registrada na época de amamentação exclusiva em relação às crianças com alimentação mista e àquelas que já não mamavam. Para ETEC, EPEC e EAggEC, a introdução de alimentos ao desmame e ao final da amamentação estiveram associadas com um aumento das infecções assintomáticas.

Diarrheagenics Escherichia coli are the major agents involved in diarrheal disease in developing countries. The aim of this study was to evaluate the time of appearance of the first asymptomatic infection by the different categories of diarrheagenic E. coli in 44 children since their birth and during the first 20 months of their lives. In all of the children studied, we detected at least one category of diarrheagenic E. coli through the 20 months of the study. 510 diarrheagenic E. coli (33.5%) were obtained from the 1,524 samples collected from the 44 children during the time of the study (31.4% EAggEC, 28.8% EPEC, 27.1% DAEC, and 12.7% ETEC). Neither EHEC nor EIEC were identified. The median age for diarrheagenic E. coli colonization was 7.5 months. The mean weaning period was 12.8 months and the mean age for introduction of mixed feeding (breast fed supplemented) was 3.8 months. A significantly lower incidence of diarrheal disease and asymptomatic infections was recorded among the exclusively breast-fed rather than in the supplemented and non breast-fed infants. For ETEC, EPEC and EAggEC the introduction of weaning foods and complete termination of breast-feeding were associated with an increase of asymptomatic infections.

Children under five years of age, from two communities of different socio-economic strata (97 from Zaiman and 55 from Las Dolores) were examined epidemiologically during 2 years, by means of quarterly visits of the working team, who carried out the collection of faecal samples. During the study, one or more enteropathogens were identified in 73.9% of samples in children from Zaiman and in 58.3% of the samples from Las Dolores, being associated to diarrhoea in 70.5% and to asymptomatic infections in 65.7%. The number of diarrheic episodes was higher in Zaiman (15.45%) than in Las Dolores (12.35%), being more frequent in the spring-summer seasons. In Zaiman, the bacterial enteropathogen proportion was relevantly higher (p< 0.005) in children with diarrhoea, whereas the presence of parasites was more frequent in asymptomatic children (p< 0.01). Rotavirus had an even distribution within diarrheic and asymptomatic children. In Las Dolores, no relevant differences were found in the detection of enteroparasites between diarrheic and asymptomatic children. Mixed infections were detected; enterotoxigenic Escherichia coli (ETEC)-rotavirus and ETEC-parasites being the most frequent ones. ETEC was involved in 85% of these infections. These data, together with the high enteropathogen carriage, suggest an elevated level of environmental contamination. The latter plays an important role in diarrheic diseases, and added to the most extreme poverty, it affects children's lives.

Niños menores de cinco años de edad de dos comunidades de diferente condición socio-económica (97 del Barrio Zaimán y 55 del Barrio Las Dolores), fueron vigilados epidemiologicamente durante 2 años, mediante visitas trimestrales del equipo de trabajo, el que realizó la toma de muestras fecales. Durante el estudio, se identificaron uno o más enteropatógenos en el 73,9% de las muestras de niños del Barrio Zaimán y en el 58,3% de Las Dolores, estando asociados a diarrea en un 70,5% y en infecciones asintomáticas en un 65,7%. El número de episodios de diarrea fue superior en Zaimán (15,5%) que en Las Dolores (12,4%), presentándose con mayor frecuencia en los meses de primavera-verano. En el Barrio Zaimán la proporción de enteropatógenos bacterianos fue significativamente mayor (p < 0.005) en los niños con diarrea, mientras que los parásitos se presentaron con mayor frecuencia en niños asintomáticos (p < 0.01). Rotavirus tuvo una distribución comparable en niños con diarrea y asintomáticos. En el Barrio Las Dolores, no se encontraron diferencias significativas en la deteccion de enteroparásitos entre asintomáticos y niños con diarrea. Se detectaron infecciones mixtas, siendo ECET-rotavirus y ECET-parásitos, las mas frecuentes. ECET estuvo involucrado en el 85%; de las mismas. Estos datos y la elevada portación de enteropatógenos, sugieren un alto nivel de contaminación ambiental, la cual juega un rol importante en la enfermedad diarreica, que sumada a la pobreza más extrema, afecta la vida de los niños