Diversidad de enterobacterias asociadas a frutos de tomate (Lycopersi-cum sculentum Mill) y suelos de invernadero

The aim of this study was to evaluate the diversity of Enterobacteriaceae present in soil and tomato fruits from three greenhouses with fertirigation system. These crop systems are an important alternative for production in protected agriculture; however, there is little information about the microbiological quality of the fruit and its relationship with chemical soil characteristics. Soil evaluations consisted of analyzing organic matter content and pH. In the microbiological analysis were isolated and identified enterobacterias organisms from composite samples of soil and fruits at different stages of maturity (0, 50 and 100%). Culture media used was selective, differential and confirmatory testing with VITEK system. Enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) were characterized genotypically, amplifying the lngA and bfpA genes by the technique of polymerase chain reaction (PCR). Diversity index (Simpson (D), Shannon-Wiener (H') and Chao estimator (SChao1) were calculated with the identified species. The species Enterobacter cloacae, Citrobacter freundii and C. brakii had a higher frequency of isolation, EPEC and ETEC were identified in soil samples and in fruits with 100% maturity. In soil, H' indices were positively correlated with the highest organic matter percentages. In fruit, although H 'and D showed less diverse bacterial communities, the isolation of ETEC and Shigella boydii on the fruit surface compromise their safety because they are usually consumed raw.El objetivo de este trabajo fue determinar la diversidad de enterobacterias presentes en suelo y tomates provenientes de tres invernaderos de fertirrigación. Estos sistemas de cultivo son una alternativa importante de producción en agricultura protegida, sin embargo, existe escasa información acerca de la calidad microbiológica de los frutos y su relación con las características químicas del suelo. Las evaluaciones del suelo consistieron en analizar el contenido de materia orgánica y pH. En los análisis microbiológicos se aislaron e identificaron enterobacterias en muestras compuestas de suelo y frutos con diferentes grados de madurez (0, 50 y 100%), utilizando medios de cultivo selectivos, diferenciales y pruebas confirmatorias con el sistema VITEK. Los patogrupos de Escherichia coli enteropatogena (EPEC) y enterotoxigénica (ETEC) se caracterizaron genotípicamente mediante la técnica de reacción en cadena de la polimerasa (PCR), amplificándose los genes bfpA y lngA. Con las especies identificadas se calcularon los índices de diversidad Simpson (D) y Shannon-Wiener (H´) y estimador de Chao (SChao1). Las especies Enterobacter cloacae, Citrobacter freundii y C. brakii presentaron mayor frecuencia de aislamiento, EPEC y ETEC fueron identificadas en muestras de suelo y en frutos con 100% de madurez. En suelo, los porcentajes de materia orgánica se correlacionaron positivamente con los índices H´. En fruto, aunque H´ y D reflejaron comunidades bacterianas menos diversas, el aislamiento de ETEC y Shigella boydii sobre la superficie del fruto comprometen su inocuidad debido a que habitualmente se consume en forma cruda

Diversidad de enterobacterias asociadas a frutos de tomate (Lycopersi-cum sculentum Mill) y suelos de invernadero

The aim of this study was to evaluate the diversity of Enterobacteriaceae present in soil and tomato fruits from three greenhouses with fertirigation system. These crop systems are an important alternative for production in protected agriculture; however, there is little information about the microbiological quality of the fruit and its relationship with chemical soil characteristics. Soil evaluations consisted of analyzing organic matter content and pH. In the microbiological analysis were isolated and identified enterobacterias organisms from composite samples of soil and fruits at different stages of maturity (0, 50 and 100%). Culture media used was selective, differential and confirmatory testing with VITEK system. Enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) were characterized genotypically, amplifying the lngA and bfpA genes by the technique of polymerase chain reaction (PCR). Diversity index (Simpson (D), Shannon-Wiener (H') and Chao estimator (SChao1) were calculated with the identified species. The species Enterobacter cloacae, Citrobacter freundii and C. brakii had a higher frequency of isolation, EPEC and ETEC were identified in soil samples and in fruits with 100% maturity. In soil, H' indices were positively correlated with the highest organic matter percentages. In fruit, although H 'and D showed less diverse bacterial communities, the isolation of ETEC and Shigella boydii on the fruit surface compromise their safety because they are usually consumed raw.El objetivo de este trabajo fue determinar la diversidad de enterobacterias presentes en suelo y tomates provenientes de tres invernaderos de fertirrigación. Estos sistemas de cultivo son una alternativa importante de producción en agricultura protegida, sin embargo, existe escasa información acerca de la calidad microbiológica de los frutos y su relación con las características químicas del suelo. Las evaluaciones del suelo consistieron en analizar el contenido de materia orgánica y pH. En los análisis microbiológicos se aislaron e identificaron enterobacterias en muestras compuestas de suelo y frutos con diferentes grados de madurez (0, 50 y 100%), utilizando medios de cultivo selectivos, diferenciales y pruebas confirmatorias con el sistema VITEK. Los patogrupos de Escherichia coli enteropatogena (EPEC) y enterotoxigénica (ETEC) se caracterizaron genotípicamente mediante la técnica de reacción en cadena de la polimerasa (PCR), amplificándose los genes bfpA y lngA. Con las especies identificadas se calcularon los índices de diversidad Simpson (D) y Shannon-Wiener (H´) y estimador de Chao (SChao1). Las especies Enterobacter cloacae, Citrobacter freundii y C. brakii presentaron mayor frecuencia de aislamiento, EPEC y ETEC fueron identificadas en muestras de suelo y en frutos con 100% de madurez. En suelo, los porcentajes de materia orgánica se correlacionaron positivamente con los índices H´. En fruto, aunque H´ y D reflejaron comunidades bacterianas menos diversas, el aislamiento de ETEC y Shigella boydii sobre la superficie del fruto comprometen su inocuidad debido a que habitualmente se consume en forma cruda

Factores ambientales que afectan la expresión del flagelo de escherichia coli enteropatógena y su implicación en la formación de biopelículas en materiales inertes

“Se sabe que las condiciones microambientales en el hospedero son un factor determinante para los microorganismos oportunistas en las primeras fases de la infección, tal es el caso de las bacterias que generan enfermedades diarreicas, las que siguen siendo un problema de salud públIica en los países en desarrollo. México ocupa uno de los primeros lugares en este tipo de enfermedades principalmente producidas por E. coli enteropatógena. Se ha asociado al flagelo de EPEC como un mediador de adherencia a células epiteliales, lo que le permite sobrevivir en el medio intestinal y ser causante de infección. En esta investigación se trabajó la interacción de EPEC en condiciones “microambientales”, encontrándose que la transcripción como la expresión del flagelo de E. coli enteropatogénica se favoreció en a) cultivos celulares, b) en presencia de desoxicolato de sodio, c) en medio D-MEM precondicionado, en presencia de algunos nutrientes y en presencia de algunos metales pesados.

Identification and Characterization of lpfABCC′DE, a Fimbrial Operon of Enterohemorrhagic Escherichia coli O157:H7

The mechanisms underlying the adherence of Escherichia coli O157:H7 and other enterohemorrhagic E. coli (EHEC) strains to intestinal epithelial cells are poorly understood. We have identified a chromosomal region (designated lpfABCC′DE) in EHEC O157:H7 containing six putative open reading frames that was found to be closely related to the long polar (LP) fimbria operon (lpf) of Salmonella enterica serovar Typhimurium, both in gene order and in conservation of the deduced amino acid sequences. We show that lpfABCC′DE is organized as an operon and that its expression is induced during the exponential growth phase. The lpf genes from EHEC strain EDL933 were introduced into a nonfimbriated (Fim(−)) E. coli K-12 strain, and the transformed strain produced fimbriae as visualized by electron microscopy and adhered to tissue culture cells. Anti-LpfA antiserum recognized a ca. 16-kDa LpfA protein when expressed under regulation of the T7 promoter system. The antiserum also cross-reacted with the LP fimbriae in immunogold electron microscopy and Western blot experiments. Isogenic E. coli O157:H7 lpf mutants derived from strains 86-24 and AGT300 showed slight reductions in adherence to tissue culture cells and formed fewer microcolonies compared with their wild-type parent strains. The adherence and microcolony formation phenotypes were restored when the lpf operon was introduced on a plasmid. We propose that LP fimbriae participate in the interaction of E. coli O157:H7 with eukaryotic cells by assisting in microcolony formation

Diversity of enterobacteria associated with tomato (Lycopersicum sculentum Mill) fruits and greenhouse soils

The aim of this study was to evaluate the diversity of Enterobacteriaceae present in soil and tomato fruits from three greenhouses with fertirigation system. These crop systems are an important alternative for production in protected agriculture; however, there is little information about the microbiological quality of the fruit and its relationship with chemical soil characteristics. Soil evaluations consisted of analyzing organic matter content and pH. In the microbiological analysis were isolated and identified enterobacterias organisms from composite samples of soil and fruits at different stages of maturity (0, 50 and 100%). Culture media used was selective, differential and confirmatory testing with VITEK system. Enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) were characterized genotypically, amplifying the lngA and bfpA genes by the technique of polymerase chain reaction (PCR). Diversity index (Simpson (D), Shannon-Wiener (H') and Chao estimator (SChao1) were calculated with the identified species. The species Enterobacter cloacae, Citrobacter freundii and C. brakii had a higher frequency of isolation, EPEC and ETEC were identified in soil samples and in fruits with 100% maturity. In soil, H' indices were positively correlated with the highest organic matter percentages. In fruit, although H 'and D showed less diverse bacterial communities, the isolation of ETEC and Shigella boydii on the fruit surface compromise their safety because they are usually consumed raw

Diversity of enterobacteria associated with tomato (Lycopersicum sculentum Mill) fruits and greenhouse soils

The aim of this study was to evaluate the diversity of Enterobacteriaceae present in soil and tomato fruits from three greenhouses with fertirigation system. These crop systems are an important alternative for production in protected agriculture; however, there is little information about the microbiological quality of the fruit and its relationship with chemical soil characteristics. Soil evaluations consisted of analyzing organic matter content and pH. In the microbiological analysis were isolated and identified enterobacterias organisms from composite samples of soil and fruits at different stages of maturity (0, 50 and 100%). Culture media used was selective, differential and confirmatory testing with VITEK system. Enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) were characterized genotypically, amplifying the lngA and bfpA genes by the technique of polymerase chain reaction (PCR). Diversity index (Simpson (D), Shannon-Wiener (H') and Chao estimator (SChao1) were calculated with the identified species. The species Enterobacter cloacae, Citrobacter freundii and C. brakii had a higher frequency of isolation, EPEC and ETEC were identified in soil samples and in fruits with 100% maturity. In soil, H' indices were positively correlated with the highest organic matter percentages. In fruit, although H 'and D showed less diverse bacterial communities, the isolation of ETEC and Shigella boydii on the fruit surface compromise their safety because they are usually consumed raw

The Transcription of Flagella of Enteropathogenic Escherichia coli O127:H6 Is Activated in Response to Environmental and Nutritional Signals

The flagella of enteropathogenic Escherichia coli (EPEC) O127:H6 E2348/69 mediate adherence to host proteins and epithelial cells. What environmental and nutritional signals trigger or down-regulate flagella expression in EPEC are largely unknown. In this study, we analyzed the influence of pH, oxygen tension, cationic and anionic salts (including bile salt), carbon and nitrogen sources, and catecholamines on the expression of the flagellin gene (fliC) of E2348/69. We found that sodium bicarbonate, which has been shown to induce the expression of type III secretion effectors, down-regulated flagella expression, explaining why E2348/69 shows reduced motility and flagellation when growing in Dulbecco’s Minimal Essential Medium (DMEM). Further, growth under a 5% carbon dioxide atmosphere, in DMEM adjusted to pH 8.2, in M9 minimal medium supplemented with 80 mM glucose or sucrose, and in DMEM containing 150 mM sodium chloride, 0.1% sodium deoxycholate, or 30 µM epinephrine significantly enhanced fliC transcription to different levels in comparison to growth in DMEM alone. When EPEC was grown in the presence of HeLa cells or in supernatants of cultured HeLa cells, high levels (4-fold increase) of fliC transcription were detected in comparison to growth in DMEM alone. Our data suggest that nutritional and host signals that EPEC may encounter in the intestinal niche activate fliC expression in order to favor motility and host colonization

Diversidad de enterobacterias asociadas a frutos de tomate (Lycopersi-cum sculentum Mill) y suelos de invernadero

The aim of this study was to evaluate the diversity of Enterobacteriaceae present in soil and tomato fruits from three greenhouses with fertirigation system. These crop systems are an important alternative for production in protected agriculture; however, there is little information about the microbiological quality of the fruit and its relationship with chemical soil characteristics. Soil evaluations consisted of analyzing organic matter content and pH. In the microbiological analysis were isolated and identified enterobacterias organisms from composite samples of soil and fruits at different stages of maturity (0, 50 and 100%). Culture media used was selective, differential and confirmatory testing with VITEK system. Enteropathogenic E. coli (EPEC) and enterotoxigenic E. coli (ETEC) were characterized genotypically, amplifying the lngA and bfpA genes by the technique of polymerase chain reaction (PCR). Diversity index (Simpson (D), Shannon-Wiener (H') and Chao estimator (SChao1) were calculated with the identified species. The species Enterobacter cloacae, Citrobacter freundii and C. brakii had a higher frequency of isolation, EPEC and ETEC were identified in soil samples and in fruits with 100% maturity. In soil, H' indices were positively correlated with the highest organic matter percentages. In fruit, although H 'and D showed less diverse bacterial communities, the isolation of ETEC and Shigella boydii on the fruit surface compromise their safety because they are usually consumed raw.El objetivo de este trabajo fue determinar la diversidad de enterobacterias presentes en suelo y tomates provenientes de tres invernaderos de fertirrigación. Estos sistemas de cultivo son una alternativa importante de producción en agricultura protegida, sin embargo, existe escasa información acerca de la calidad microbiológica de los frutos y su relación con las características químicas del suelo. Las evaluaciones del suelo consistieron en analizar el contenido de materia orgánica y pH. En los análisis microbiológicos se aislaron e identificaron enterobacterias en muestras compuestas de suelo y frutos con diferentes grados de madurez (0, 50 y 100%), utilizando medios de cultivo selectivos, diferenciales y pruebas confirmatorias con el sistema VITEK. Los patogrupos de Escherichia coli enteropatogena (EPEC) y enterotoxigénica (ETEC) se caracterizaron genotípicamente mediante la técnica de reacción en cadena de la polimerasa (PCR), amplificándose los genes bfpA y lngA. Con las especies identificadas se calcularon los índices de diversidad Simpson (D) y Shannon-Wiener (H´) y estimador de Chao (SChao1). Las especies Enterobacter cloacae, Citrobacter freundii y C. brakii presentaron mayor frecuencia de aislamiento, EPEC y ETEC fueron identificadas en muestras de suelo y en frutos con 100% de madurez. En suelo, los porcentajes de materia orgánica se correlacionaron positivamente con los índices H´. En fruto, aunque H´ y D reflejaron comunidades bacterianas menos diversas, el aislamiento de ETEC y Shigella boydii sobre la superficie del fruto comprometen su inocuidad debido a que habitualmente se consume en forma cruda