Fast and easy detection of CMY-2 in Escherichia coli by direct MALDI-TOF mass spectrometry.

Fast typing methods for third generation cephalosporin resistance mechanisms are needed to guide appropriate treatment and prevent potential dissemination events. In this study we used a novel short and fast methodology for the identification of CMY-2 in 50 well characterized clinical isolates of E. coli by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry- MALDI-TOF MS. Samples were prepared using the double layer sinapinic acid technique for detection of intact proteins Comparison among mass spectral profile of different strains between m/z 35,000-45,000 Da showed that two groups of isolates could be differentiated after peak analysis. A single distinctive peak with different intensities, at approximately m/z 39,800 Da was found in all CMY-2 producing strains (transconjugant, transformant and wild type) and consistently absent in the control groups (ESBL producers and susceptible strains). Statistical results showed 100% values for sensitivity and specificity, indicating a perfect test and a high discriminative power. In this study, we demonstrated that MALDI-TOF MS has the potential to detect directly the most clinically relevant acquired AmpC ß-lactamase, the CMY-2-enzyme, in E. coli with a less time-consuming process as compared to conventional methods. Our results may constitute the basis for further research to detect other ß-lactamases, or even other resistance markers.

Clonal relations of atypical enteropathogenic Escherichia coli O157:H16 strains isolated from various sources from several countries.

Atypical enteropathogenic Escherichia coli (aEPEC) is comprised of a large heterogeneous group of strains and serotypes that carry the intimin gene (eae) but no other EPEC virulence factors. In a previous study, we examined a few aEPEC strains of O157:H16 serotype from the U.S. and France and found these to be nearly homologous, and speculated that the same strain had been disseminated or perhaps they are part of a large clonal group that exists worldwide. To test that hypothesis, we examined additional 45 strains isolated from various sources from 4 other countries and determined that although there are a few eae-negative O157:H16 strains, most are aEPEC that carried eae and specifically, the ε-eae allele. Analysis by pulsed field gel electrophoresis (PFGE) and multilocus sequence typing showed that as a whole, O157:H16 strains are phylogenetically diverse and have different sequence types and PFGE profiles. But the aEPEC strains within the O157:H16 serotype, regardless of the eae allele carried, are a highly conserved and homologous group of sequence type (ST)-171 strains that shared similar PFGE profiles. These aEPEC strains of O157:H16 serotype are not closely related to any of the major EPEC and enterohemorrhagic E. coli clonal lineages and appear to be part of a large clonal group that are prevalent worldwide.

Escherichia coli shigatoxigénica en animales relacionados con casos de diarreas sanguinolentas o síndrome urémico hemolítico y prevalencia en roedores de la Ciudad de Buenos Aires / Shiga toxin-producing escherichia coli in animals related to cases of bloody diarrhea

Escherichia coli shigatoxigénica (STEC) es un patógeno endémico en Argentina, responsable de diarrea aguda sanguinolenta (DAS) y/o síndrome urémico hemolítico (SUH). La correlación entre SUH y alimentos contaminados ha sido documentada, aunque no siempre se estableció la fuente de infección. La ruta de contagio persona-persona es relevante. Dados los registros previos de prevalencia de STEC en animales de compañía y los hábitos de convivencia humano-animal en centros urbanos, es necesario evaluar la ruta mascota-persona. A su vez, los roedores podrían tener un papel epidemiológico en la endemia. OBJETIVO: Estudiar posibles reservorios animales relacionados con casos de SUH/DAS en la Ciudad Autónoma de Buenos Aires y estimar la prevalencia de STEC en roedores. MÉ-TODOS: Se intervino en 28 casos de SUH y 49 de DAS. Se realizó rastrillaje de cepas STEC por PCR a partir de hisopados rectales de los animales vinculados a cada caso. La prevalencia en roedores se estimó por PCR de sus hisopados rectales. RESULTADOS: Se aislaron cepas STEC en 1/10 caninos y 1/3 felinos con vivientes con casos de SUH, y 1/9 felinos contacto con casos de DAS. Rattus rattus fue hospedero de cepas STEC en 33% de los animales capturados en focos de SUH. En roedores, la prevalencia fue de 3,1%. CONCLUSIONES: Las cepas STEC circulan en los animales que conviven o tienen al menos un hábitat compartido con la población en riesgo, quienes podrían participar en la transmisión del agente. Es necesario reevaluar las intervenciones sanitarias en focos y en programas de control de SUH/DAS

Shigatoxigenic Escherichia coli(STEC) is an endemic pathogen in Argentina, which causes bloody diarrhea (BD) and/or hemolytic uremic syndrome (HUS).The co-relation between HUS and contaminated food has been documented, although the source of infection was not always established. Person-to-person route of infection is relevant. Taking into account previous STEC prevalence data in companion animals and the habits of human-animal coexistence in urban centers, it is necessary to evaluate pet-to-person transmission. On the other hand, rodents may also play an epidemiologic role in the endemic transmission. OBJECTIVE: To study potential animal reservoirs related to HUS and BD cases in the City of Buenos Aires and to estimate the prevalence of STEC in rodents. METHODS: An intervention was conducted in 28 cases of HUS and 49 of BD. Screening for STEC was performed by PCR from rectal swabs of linked animals to each case. The prevalence in rodents was estimated by PCR from rectal swabs. RESULTS: STEC strains were isolated in 1/10 dogs and 1/3 cats cohabiting with HUS cases, and in 1/9 cats in contact with DAS cases. Rattus rattus was host of STEC strains in 33% of the animals captured in HUS areas. In rodents, the prevalence was 3.1%. CONCLUSIONS: STEC strains circulate in animals that live withor share at least the same habitat with the population at risk, and could participate in the transmission of the agent. It is necessary to re-evaluate health interventions both in outbreaks and in control programs of HUS/BD

Escherichia coli shigatoxigénica en animales relacionados con casos de diarreas sanguinolentas o síndrome urémico hemolítico y prevalencia en roedores de la Ciudad de Buenos Aires / Shiga toxin-producing escherichia coli in animals related to cases of bloody diarrhea

Escherichia coli shigatoxigénica (STEC) es un patógeno endémico en Argentina, responsable de diarrea aguda sanguinolenta (DAS) y/o síndrome urémico hemolítico (SUH). La correlación entre SUH y alimentos contaminados ha sido documentada, aunque no siempre se estableció la fuente de infección. La ruta de contagio persona-persona es relevante. Dados los registros previos de prevalencia de STEC en animales de compañía y los hábitos de convivencia humano-animal en centros urbanos, es necesario evaluar la ruta mascota-persona. A su vez, los roedores podrían tener un papel epidemiológico en la endemia. OBJETIVO: Estudiar posibles reservorios animales relacionados con casos de SUH/DAS en la Ciudad Autónoma de Buenos Aires y estimar la prevalencia de STEC en roedores. ME-TODOS: Se intervino en 28 casos de SUH y 49 de DAS. Se realizó rastrillaje de cepas STEC por PCR a partir de hisopados rectales de los animales vinculados a cada caso. La prevalencia en roedores se estimó por PCR de sus hisopados rectales. RESULTADOS: Se aislaron cepas STEC en 1/10 caninos y 1/3 felinos con vivientes con casos de SUH, y 1/9 felinos contacto con casos de DAS. Rattus rattus fue hospedero de cepas STEC en 33% de los animales capturados en focos de SUH. En roedores, la prevalencia fue de 3,1%. CONCLUSIONES: Las cepas STEC circulan en los animales que conviven o tienen al menos un hábitat compartido con la población en riesgo, quienes podrían participar en la transmisión del agente. Es necesario reevaluar las intervenciones sanitarias en focos y en programas de control de SUH/DAS (AU)

Shigatoxigenic Escherichia coli(STEC) is an endemic pathogen in Argentina, which causes bloody diarrhea (BD) and/or hemolytic uremic syndrome (HUS).The co-relation between HUS and contaminated food has been documented, although the source of infection was not always established. Person-to-person route of infection is relevant. Taking into account previous STEC prevalence data in companion animals and the habits of human-animal coexistence in urban centers, it is necessary to evaluate pet-to-person transmission. On the other hand, rodents may also play an epidemiologic role in the endemic transmission. OBJECTIVE: To study potential animal reservoirs related to HUS and BD cases in the City of Buenos Aires and to estimate the prevalence of STEC in rodents. METHODS: An intervention was conducted in 28 cases of HUS and 49 of BD. Screening for STEC was performed by PCR from rectal swabs of linked animals to each case. The prevalence in rodents was estimated by PCR from rectal swabs. RESULTS: STEC strains were isolated in 1/10 dogs and 1/3 cats cohabiting with HUS cases, and in 1/9 cats in contact with DAS cases. Rattus rattus was host of STEC strains in 33% of the animals captured in HUS areas. In rodents, the prevalence was 3.1%. CONCLUSIONS: STEC strains circulate in animals that live withor share at least the same habitat with the population at risk, and could participate in the transmission of the agent. It is necessary to re-evaluate health interventions both in outbreaks and in control programs of HUS/BD (AU)