Whole genome sequencing of Shigella sonnei through PulseNet Latin America and Caribbean: advancing global surveillance of foodborne illnesses.

OBJECTIVES: Shigella sonnei is a globally important diarrhoeal pathogen tracked through the surveillance network PulseNet Latin America and Caribbean (PNLA&C), which participates in PulseNet International. PNLA&C laboratories use common molecular techniques to track pathogens causing foodborne illness. We aimed to demonstrate the possibility and advantages of transitioning to whole genome sequencing (WGS) for surveillance within existing networks across a continent where S. sonnei is endemic. METHODS: We applied WGS to representative archive isolates of S. sonnei (n = 323) from laboratories in nine PNLA&C countries to generate a regional phylogenomic reference for S. sonnei and put this in the global context. We used this reference to contextualise 16 S. sonnei from three Argentinian outbreaks, using locally generated sequence data. Assembled genome sequences were used to predict antimicrobial resistance (AMR) phenotypes and identify AMR determinants. RESULTS: S. sonnei isolates clustered in five Latin American sublineages in the global phylogeny, with many (46%, 149 of 323) belonging to previously undescribed sublineages. Predicted multidrug resistance was common (77%, 249 of 323), and clinically relevant differences in AMR were found among sublineages. The regional overview showed that Argentinian outbreak isolates belonged to distinct sublineages and had different epidemiologic origins. CONCLUSIONS: Latin America contains novel genetic diversity of S. sonnei that is relevant on a global scale and commonly exhibits multidrug resistance. Retrospective passive surveillance with WGS has utility for informing treatment, identifying regionally epidemic sublineages and providing a framework for interpretation of prospective, locally sequenced outbreaks.

Bacterial pathogens associated with bloody diarrhea in Uruguayan children.

Diarrheal disease continues to be a serious health problem, especially in developing countries. Bloody diarrhea represents approximately 20-30% of all cases and has higher morbidity and mortality. Treatment with antibiotics is beneficial in cases of Shigella, Campylobacter, Yersinia and Salmonella infection, principally in those children with a higher risk of invasive disease. The aims of this study were to detect the bacterial agents associated with bloody diarrhea in children and to determine their antimicrobial susceptibility patterns. Between June 2001 and January 2008, 249 children with bloody diarrhea were studied. Shigella and Shiga toxin-producing Escherichia coli (STEC) were recovered from 48 (19.3%) and 3 (1.2%) of the total of cases, respectively. In 49 out of 249 children, in whom other enteropathogens were investigated, we recovered Campylobacter jejuni from 7 children (14.3%), Salmonella spp. from 2 (4.1%) and Aeromonas spp. from 1 (2%) in addition to Shigella from 7 children (14.3%). Thirty-four (70%) Shigella isolates showed resistance to ampicillin and 13 (27%) to trimethoprim-sulfamethoxazole. All Shigella isolates were susceptible to nalidixic acid, ciprofloxacin and ceftriaxone. Salmonella and STEC isolates were susceptible to all antibiotics assayed. Thus, the use of trimethoprim-sulfamethoxazole or ampicillin would not be appropriate for the empirical treatment of Shigella - associated diarrhea.

Comparative molecular study of Mycobacterium tuberculosis strains, in times of antimicrobial drug resistance.

Strains of Mycobacterium tuberculosis were compared using two DNA fingerprinting techniques: Restriction Fragment Length Polymorphism (RFLP) and Double-Repetitive-Element-PCR (DRE-PCR). Two of these strains: IH1 (susceptible to isoniazid) and IH2 (resistant to isoniazid) were recovered from cases of pulmonary tuberculosis which occurred in two brothers who lived together. The first one was recognized on July 1999, and the second was diagnosed one year later. IH1 and IH2 showed the same pattern of bands with both molecular tests. These results suggest that single drug chemoprophylaxis may occasionally select resistant strains for that drug, which can eventually cause disease and be recognized through these tests. Strains IH3, IH4 and IH5 were obtained from sputum samples of 3 different patients, and intra-laboratory cross-contamination was suspected when it was realized that the 3 positive materials had been consecutively processed the same day by the same worker in the same biological safety cabinet. Again, the 3 strains revealed identical band patterns with RFLP and DRE-PCR, confirming the posed suspicion. The results with DRE-PCR were obtained after only 8 hours of work, without the need for subcultures. This procedure allows quick correction of treatment conducts, avoiding unnecessary exposure of people and bacteria to antimicrobial drugs.

Comparative molecular study of Mycobacterium tuberculosis strains, in times of antimicrobial drug resistance / Estudio molecular comparativo de cepas de Mycobacterium tuberculosis, en tiempos de resistencia antimicrobiana a los fármacos

Strains of Mycobacterium tuberculosis were compared using two DNA fingerprinting techniques: Restriction Fragment Length Polymorphism (RFLP) and Double-Repetitive-Element-PCR (DRE-PCR). Two of these strains: IH1 (susceptible to isoniazid) and IH2 (resistant to isoniazid) were recovered from cases of pulmonary tuberculosis which occurred in two brothers who lived together. The first one was recognized on July 1999, and the second was diagnosed one year later. IH1 and IH2 showed the same pattern of bands with both molecular tests. These results suggest that single drug chemoprophylaxis may occasionally select resistant strains for that drug, which can eventually cause disease and be recognized through these tests. Strains IH3, IH4 and IH5 were obtained from sputum samples of 3 different patients, and intra-laboratory cross-contamination was suspected when it was realized that the 3 positive materials had been consecutively processed the same day by the same worker in the same biological safety cabinet. Again, the 3 strains revealed identical band patterns with RFLP and DRE-PCR, confirming the posed suspicion. The results with DRE-PCR were obtained after only 8 hours of work, without the need for subcultures. This procedure allows quick correction of treatment conducts, avoiding unnecessary exposure of people and bacteria to antimicrobial drugs.

Se compararon cepas de Mycobacterium tuberculosis utilizando 2 procedimientos de ADN fingerprinting: polimorfismo de los fragmentos de restricción (RFLP) y Double-Repetitive-Element-PCR (DRE-PCR). Dos de las cepas: IH1 (susceptible a isoniazida) e IH2 (resistente a isoniazida) se recuperaron a partir de casos de tuberculosis pulmonar que ocurrieron en dos hermanos convivientes. La primera fue aislada en julio de 1999 y la segunda un año después. IH1 e IH2 mostraron el mismo patrón de bandas por ambos procedimientos. Estos resultados sugieren que la quimioprofilaxis con una sola droga puede ocasionalmente seleccionar mutantes resistentes, las cuales pueden causar enfermedad y ser reconocidas por estos procedimientos. Las cepas IH3, IH4 e IH5 fueron aisladas de 3 pacientes diferentes, y examinadas por probable contaminación cruzada dentro del laboratorio ya que fueron procesadas el mismo día, por el mismo operador y en la misma cabina de seguridad biológica. Nuevamente, las 3 cepas revelaron el mismo patrón de bandas por RFLP y por DRE-PCR, confirmando la sospecha. Los resultados de la DRE-PCR se obtuvieron luego de 8 horas de trabajo, sin necesidad de subcultivos. Esta técnica permitiría la rápida correción de pautas de tratamiento, evitando la exposición innecesaria de personas y bacterias a drogas antimicrobianas.

Serotipos, perfil plasmídico y antibiotipos de cepas de Shigella flexneri aisladas de niños menores de 5 años con diarrea sanguinolenta usuarios de los servicios de Salud Pública / Serotypes, serum profile and anti-biotypes of Shigella flexneri cultures isolated from infants under 5 years with diarrhea sanguinolenta users of Public Health Services

Introducción: en niños menores de 5 años con diarrea sanguinolenta, Shigella flexneri es el agente bacteriano más frecuentemente recuperado en nuestro medio. La shigellosis es una de las enfermedades infecciosas en las cuales el tratamiento con antimicrobianos es efectivo. La elección empírica del antimicrobiano adecuado es problemática debido a la resistencia de Shigella a diversos antibióticos. Esta situación estimuló el interés en el desarrollo de vacunas para el control de esta enfermedad. Debido a que algunas vacunas están orientadas a promover la respuesta inmune serotipo específica, es importante establecer la distribución de los serotipos prevalentes en la población que interesa inmunizar. El objetivo de este estudio fue caracterizar 50 cepas de Shigella flexneri aisladas a partir de niños con diarrea sanguinolenta, recuperadas en 4 encuestas etiológicas de gastroenteritis. Método: a cada una se le realizó: serotipificación, estudio del patrón de lipopolisacárido, perfil plasmídico y estudio de sensibilidad a diferentes antimicrobianos. Resultados: los seroptipos prevalentes fueron 2a, 3c, 4, 6, y 1. Se identificaron 10 antibotipos diferentes. En los cultivos del serotipo 2a se hallaron 3 patrones plasmídicos; el 5 fue el más frecuente, seguido por el 6 y el 7. El análisis de la evolución de los antibiotipos circulantes mostró una tendencia hacia la aparición de tipos con mayor espectro de resistencia. Conclusiones: vista esta evolución, y de acuerdo a los resultados obtenidos, resulta de interés ensayar un inmunógeno que incluya los determinantes "O" específicos de los serotipor 2a, 1, 3c, 4, 6 y el de Shigella sonnei.

Comparative molecular study of Mycobacterium tuberculosis strains, in times of antimicrobial drug resistance.

Strains of Mycobacterium tuberculosis were compared using two DNA fingerprinting techniques: Restriction Fragment Length Polymorphism (RFLP) and Double-Repetitive-Element-PCR (DRE-PCR). Two of these strains: IH1 (susceptible to isoniazid) and IH2 (resistant to isoniazid) were recovered from cases of pulmonary tuberculosis which occurred in two brothers who lived together. The first one was recognized on July 1999, and the second was diagnosed one year later. IH1 and IH2 showed the same pattern of bands with both molecular tests. These results suggest that single drug chemoprophylaxis may occasionally select resistant strains for that drug, which can eventually cause disease and be recognized through these tests. Strains IH3, IH4 and IH5 were obtained from sputum samples of 3 different patients, and intra-laboratory cross-contamination was suspected when it was realized that the 3 positive materials had been consecutively processed the same day by the same worker in the same biological safety cabinet. Again, the 3 strains revealed identical band patterns with RFLP and DRE-PCR, confirming the posed suspicion. The results with DRE-PCR were obtained after only 8 hours of work, without the need for subcultures. This procedure allows quick correction of treatment conducts, avoiding unnecessary exposure of people and bacteria to antimicrobial drugs.

Primer aislamiento en Uruguay de Escherichia Coli, productora de toxina Shiga del serotipo O157: H7 en una niña con síndrome urémico hemolítico / First Uruguayan isolation of Shiga toxin-producing Escherichia coli O157: H7 in female infant with hemolytic syndrome

En mayo de 2002 se aisló por primera vez en Uruguay Escherichia coli O157:H7, productora de toxina shiga a partir del coproductivo de una niña de 16 meses procedente de Melo, con diagnóstico de síndrome urémico hemolítico. La cepa, productora de toxinas shiga tipo 2 y tipo 2 variante humana a, era genéticamente distinta de las cepas circulantes en Argentina.

Efecto del Lactobacillus casei administrado en el suero de rehidratación oral, en el tratamiento de la enfermedad diarreica aguda / Efficacy of Lactobacillus GG in oral rehidratation solution

El probiótico Lactobacillus GG es efectivo en promover una recuperación rápida de la diarrea aguda infantil producida por rotavirus. Hay poca información, sobre el rol de este agente en el efecto sobre la diarrea producida por otros gérmenes, tampoco hay evidencia de su eficacia administrada en la sal de hidratación oral para pscientes con diarrea de diversas causa. Método: Niños de 1 mes a 3 años de edad con enfermedad diarreica aguda fueron enrolados en una investigación doble ciego. Pacientes fueron randomizados y colocados en al grupo A recibiendo sal de hidratación y placebo, y en el grupo recibiendo, el mismo con el agragado de una preparación viva de Lactobacuillus GG. Luego de la hidratación en las 4 ó 6 horas se les ofreció su dieta habitual. Resultados: enrolamos 97 pacientes 52 del grupo A y 45 del grupo B. Duración de la diarrea luego del enrolamiento fue 7 días para el grupo A y 4 días para el grupo B (p<0.005).Para los niños rotavirus positivos la diarrea duró 6 días para el grupo A y 3 días para el grupo B (p<0.005).La diarrea duró más de 10 días, 5 para el grupo A y 1 para el grupo B. Conclusión: Administrar sal de hidratación oral conteniendo Lactobacillus GG a niños con enfermedad diarreica aguda es segura y resulta en duración menor de esta y tiene menos chance de pasar a curso prolongado. (AU)