RESUMEN
The current study aimed to detect virulence, hetero-pathogenicity, and hybridization genes in Escherichia coli strains, previously isolated from cloacal swabs in commercial breeding psittacines and zoological collections, via multiplex PCR. A total of 68 strains of E. coli, previously isolated from psittacines in zoos and commercial breeding facilities in Ceará, Brazil, were assessed for the presence of the following genes and/or probes: eae, bfpA (EPEC - Enteropathogenic E. coli), CVD432 (EAEC - Enteroaggregative E. coli); LT gene and ST gene (ETEC - Enterotoxigenic E. coli); ipaH (EIEC - Enteroinvasive E. coli); stx1 and stx2 (STEC - Shiga toxin-producing E. coli); iroN, ompT, hlyF, iss, and iutA (APEC - Avian pathogenic E. coli). Of the 68 E. coli strains analyzed, 61 (98.7â¯%) were positive for the following genes and/or probes: Stx1 (61/98.7â¯%), ST gene (54/79.4â¯%), CVD432 (49/72â¯%), bfpA (44/64.7â¯%), eae (42/61.8â¯%), Stx2 (41/60.3â¯%), ipaH (34/50â¯%), LT gene (33/48.5â¯%), iroN (21/30.9â¯%), hlyF (11/6.2â¯%), iss (06/8.8â¯%) and iutA (06/8.8â¯%). The following diarrheagenic pathotypes were identified: 66 (97â¯%) from STEC, 49 (72â¯%) from EAEC, 35 (52â¯%) from EIEC, 25 (37â¯%) from ETEC, and one (1.5â¯%) from EPEC. Regarding hetero-pathogenicity, 50 (74â¯%) heterogeneous strains were identified. Positivity for APEC was seen in four (6â¯%) strains, all characterized as pathogenic hybrids. This study describes significant associations of virulence factors in E. coli strains DEC/DEC and DEC/APEC, which were isolated from psittacines and may be potentially harmful to One Health.
Asunto(s)
Animales de Zoológico , Enfermedades de las Aves , Infecciones por Escherichia coli , Proteínas de Escherichia coli , Escherichia coli , Factores de Virulencia , Animales , Brasil , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/veterinaria , Animales de Zoológico/microbiología , Enfermedades de las Aves/microbiología , Escherichia coli/genética , Escherichia coli/patogenicidad , Escherichia coli/aislamiento & purificación , Escherichia coli/clasificación , Proteínas de Escherichia coli/genética , Factores de Virulencia/genética , Virulencia/genética , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/patogenicidad , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Enteropatógena/clasificación , Reacción en Cadena de la Polimerasa Multiplex , Psittaciformes/microbiología , Cloaca/microbiología , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Escherichia coli Shiga-Toxigénica/patogenicidad , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Enterotoxigénica/genética , Escherichia coli Enterotoxigénica/patogenicidad , Escherichia coli Enterotoxigénica/aislamiento & purificación , Escherichia coli Enterotoxigénica/clasificaciónRESUMEN
Abstract This study assessed the inhibitory potential of the probiotics Lactobacillus (LB) exopolysaccharides (EPS) with or without extracts of Satureja calamintha on enteropathogenic Escherichia coli (EPEc) responsible for gastroenteritis. Methanolic and hydromethanolic extracts were prepared by cold maceration and subjected to phytochemical screening. The compounds of the extracts were determined with the colorimetric assays and identified using high-performance liquid chromatography coupled with diode array detector (HPLC-DAD). Antioxidant activities of the extracts were also evaluated by using 2,2-diphenyl-1-picrylhydrazil (DPPH) radical scavenging. Antibacterial effect on EPEc was evaluated by using both agar disc diffusion and microdilution methods. The in vitro test of auto-aggregation was investigated. Microbiological analysis showed that 63% of the isolated LB were producing EPS, with the amount ranging from 8.21 to 43.13 mg/L. Chemical analysis of the extracts revealed the presence of polyphenols and flavonoids, more abundant in the hydromethanolic extract, which presented the highest content with 2.11 mg EGA/g of polyphenol and 1.64 mg EC/g of flavonoids and 1.71 mg EGA/g of polyphenol and 1.15 mg EC/g of flavonoids in the methanolic extract. Hydromethanolic extracts and EPS exhibited a more important activity than did the methanolic extract against EPEc. The combined action of EPS and extracts reduced the aggregation ability of EPEc and decreased the rate of their adhesion.
Asunto(s)
Probióticos/efectos adversos , Satureja/efectos adversos , Escherichia coli Enteropatógena/clasificación , Lactobacillus/clasificación , Extractos Vegetales/análisis , Cromatografía Líquida de Alta Presión/métodos , Nepeta/efectos adversos , Fitoquímicos , Gastroenteritis , Antioxidantes/farmacologíaRESUMEN
Enteropathogenic (EPEC) and enteroaggregative (EAEC) Escherichia coli are two of the major pathotypes of diarrheagenic E. coli causing disease worldwide. Here, we report a diarrheal outbreak caused by E. coli of serotype O3:H2, harboring virulence markers from EPEC (eae) and/or EAEC (aggR). This is likely the first E. coli diarrheal outbreak caused by a hybrid atypical-EPEC/EAEC clone reported in Brazil.
Asunto(s)
Escherichia coli Enteropatógena , Infecciones por Escherichia coli , Brasil/epidemiología , Células Clonales , Diarrea/epidemiología , Diarrea/etiología , Diarrea/microbiología , Brotes de Enfermedades , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Enteropatógena/patogenicidad , Infecciones por Escherichia coli/complicaciones , Infecciones por Escherichia coli/epidemiología , Humanos , Serogrupo , Factores de VirulenciaAsunto(s)
Diarrea/historia , Diarrea/microbiología , Escherichia coli Enteropatógena , Infecciones por Escherichia coli/complicaciones , Infecciones por Escherichia coli/historia , Diarrea/epidemiología , Diarrea/terapia , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/terapia , Salud Global , Historia del Siglo XX , Historia del Siglo XXI , HumanosRESUMEN
BACKGROUND: Atypical enteropathogenic Escherichia coli (aEPEC) are one of the most frequent intestinal E. coli pathotypes isolated from diarrheal patients in Brazil. Isolates of aEPEC contain the locus of enterocyte effacement, but lack the genes of the bundle-forming pilus of typical EPEC, and the Shiga toxin of enterohemorrhagic E. coli (EHEC). The objective of this study was to evaluate the phylogeny and the gene content of Brazilian aEPEC genomes compared to a global aEPEC collection. METHODOLOGY: Single nucleotide polymorphism (SNP)-based phylogenomic analysis was used to compare 106 sequenced Brazilian aEPEC with 221 aEPEC obtained from other geographic origins. Additionally, Large-Scale BLAST Score Ratio was used to determine the shared versus unique gene content of the aEPEC studied. PRINCIPAL FINDINGS: Phylogenomic analysis demonstrated the 106 Brazilian aEPEC were present in phylogroups B1 (47.2%, 50/106), B2 (23.6%, 25/106), A (22.6%, 24/106), and E (6.6%, 7/106). Identification of EPEC and EHEC phylogenomic lineages demonstrated that 42.5% (45/106) of the Brazilian aEPEC were in four of the previously defined lineages: EPEC10 (17.9%, 19/106), EPEC9 (10.4%, 11/106), EHEC2 (7.5%, 8/106) and EPEC7 (6.6%, 7/106). Interestingly, an additional 28.3% (30/106) of the Brazilian aEPEC were identified in five novel lineages: EPEC11 (14.2%, 15/106), EPEC12 (4.7%, 5/106), EPEC13 (1.9%, 2/106), EPEC14 (5.7%, 6/106) and EPEC15 (1.9%, 2/106). We identified 246 genes that were more frequent among the aEPEC isolates from Brazil compared to the global aEPEC collection, including espG2, espT and espC (P<0.001). Moreover, the nleF gene was more frequently identified among Brazilian aEPEC isolates obtained from diarrheagenic patients when compared to healthy subjects (69.7% vs 41.2%, P<0.05). CONCLUSION: The current study demonstrates significant genomic diversity among aEPEC from Brazil, with the identification of Brazilian aEPEC isolates to five novel EPEC lineages. The greater prevalence of some virulence genes among Brazilian aEPEC genomes could be important to the specific virulence strategies used by aEPEC in Brazil to cause diarrheal disease.
Asunto(s)
Hibridación Genómica Comparativa/métodos , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Genoma Bacteriano , Filogenia , Factores de Virulencia/genética , Brasil , Infecciones por Escherichia coli , Proteínas de Escherichia coli/genética , Humanos , Tipificación de Secuencias Multilocus , Serotipificación , VirulenciaRESUMEN
ABSTRACT Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.
Asunto(s)
Humanos , Animales , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Peces/microbiología , Filogenia , Contaminación de Alimentos/análisis , Seguridad de Productos para el Consumidor , Proteínas de Escherichia coli/genética , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Carne/microbiologíaRESUMEN
Shigatoxigenic and enteropathogenic Escherichia coli with virulence and multidrug resistance profile were isolated from Nile tilapia. This study finding is of great importance to public health because they help understand this pathogen epidemiology in fish and demonstrate how these animals can transmit E. coli related diseases to humans.
Asunto(s)
Escherichia coli Enteropatógena/aislamiento & purificación , Peces/microbiología , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Animales , Seguridad de Productos para el Consumidor , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Proteínas de Escherichia coli/genética , Contaminación de Alimentos/análisis , Humanos , Carne/microbiología , Filogenia , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genéticaRESUMEN
BACKGROUND: Diarrheagenic Escherichia coli (DEC) are important bacterial causes of childhood diarrhea in Brazil, but its impact in adults is unknown. This study aimed at investigating DEC among children and adults living in endemic areas. METHODS: A total of 327 stools specimens were collected from children (n = 141) and adults (n = 186) with diarrhea attending health centers. Diarrheagenic E. coli (DEC) were identified by their virulence genes (multiplex polymerase chain reaction) and HEp-2 cell adherence patterns. RESULTS: DEC were detected in 56 (40%) children and 74 (39%) adults; enteroaggregative E. coli (EAEC) (23%) was the most prevalent pathotype, followed by diffusely adherent E. coli (DAEC) (13%), and occurred at similar frequencies in both diarrheal groups. Atypical enteropathogenic E. coli (aEPEC) strains were recovered more frequently from children (6%) than from adults (1%). Twenty-six percent of the EAEC were classified as typical EAEC possessing aggR gene, and carried the aap gene. EAEC strains carrying aggR-aap-aatA genes were significantly more frequent among children than adults (p < 0.05). DAEC strains possessing Afa/Dr. genes were detected from children (10%) and adults (6%). EAEC and DAEC strains harboring genes for the EAST1 (astA), Pet, Pic, and Sat toxins were common in both diarrheal groups. The astA and the porcine AE/associated adhesin (paa) genes were found in most of aEPEC strains. High levels of resistance to antimicrobial drugs were found among DAEC and aEPEC isolates. CONCLUSION: The results show a high proportion of EAEC and DAEC carrying toxin-encoding genes among adults with diarrhea.
Asunto(s)
Diarrea/microbiología , Escherichia coli Enteropatógena/aislamiento & purificación , Infecciones por Escherichia coli/microbiología , Adolescente , Adulto , Animales , Toxinas Bacterianas/metabolismo , Brasil/epidemiología , Niño , Preescolar , Diarrea/epidemiología , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Infecciones por Escherichia coli/epidemiología , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Femenino , Humanos , Masculino , Reacción en Cadena de la Polimerasa Multiplex , Prevalencia , Porcinos , Enfermedades de los Porcinos/epidemiología , Enfermedades de los Porcinos/microbiología , Factores de Virulencia/genética , Factores de Virulencia/metabolismo , Adulto JovenRESUMEN
ABSTRACT Psittacine birds have been identified as reservoirs of diarrheagenic Escherichia coli, a subset of pathogens associated with mortality of children in tropical countries. The role of other orders of birds as source of infection is unclear. The aim of this study was to perform the molecular diagnosis of infection with diarrheagenic E. coli in 10 different orders of captive wild birds in the state of São Paulo, Brazil. Fecal samples were analyzed from 516 birds belonging to 10 orders: Accipitriformes, Anseriformes, Columbiformes, Falconiformes, Galliformes, Passeriformes, Pelecaniformes, Piciformes, Psittaciformes and Strigiformes. After isolation, 401 E. coli strains were subjected to multiplex PCR system with amplification of genes eae and bfp (EPEC), stx1 and stx2 for STEC. The results of these tests revealed 23/401 (5.74%) positive strains for eae gene, 16/401 positive strains for the bfp gene (3.99%) and 3/401 positive for stx2 gene (0.75%) distributed among the orders of Psittaciformes, Strigiformes and Columbiformes. None of strains were positive for stx1 gene. These data reveal the infection by STEC, typical and atypical EPEC in captive birds. The frequency of these pathotypes is low and restricted to few orders, but the data suggest the potential public health risk that these birds represent as reservoirs of diarrheagenic E. coli.
Asunto(s)
Animales , Aves/microbiología , Reservorios de Enfermedades/microbiología , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Animales Salvajes/microbiología , Aves/clasificación , Brasil , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/metabolismo , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/metabolismo , Animales Salvajes/clasificaciónRESUMEN
Psittacine birds have been identified as reservoirs of diarrheagenic Escherichia coli, a subset of pathogens associated with mortality of children in tropical countries. The role of other orders of birds as source of infection is unclear. The aim of this study was to perform the molecular diagnosis of infection with diarrheagenic E. coli in 10 different orders of captive wild birds in the state of São Paulo, Brazil. Fecal samples were analyzed from 516 birds belonging to 10 orders: Accipitriformes, Anseriformes, Columbiformes, Falconiformes, Galliformes, Passeriformes, Pelecaniformes, Piciformes, Psittaciformes and Strigiformes. After isolation, 401 E. coli strains were subjected to multiplex PCR system with amplification of genes eae and bfp (EPEC), stx1 and stx2 for STEC. The results of these tests revealed 23/401 (5.74%) positive strains for eae gene, 16/401 positive strains for the bfp gene (3.99%) and 3/401 positive for stx2 gene (0.75%) distributed among the orders of Psittaciformes, Strigiformes and Columbiformes. None of strains were positive for stx1 gene. These data reveal the infection by STEC, typical and atypical EPEC in captive birds. The frequency of these pathotypes is low and restricted to few orders, but the data suggest the potential public health risk that these birds represent as reservoirs of diarrheagenic E. coli.
Asunto(s)
Animales Salvajes/microbiología , Aves/microbiología , Reservorios de Enfermedades/microbiología , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Animales , Animales Salvajes/clasificación , Aves/clasificación , Brasil , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/metabolismo , Proteínas de Escherichia coli/genética , Proteínas de Escherichia coli/metabolismo , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/metabolismoRESUMEN
This study aimed to detect virulence factors, pathovars, and phylogenetic groups of Escherichia coli strains obtained from feces of calves with and without diarrhea up to 70 days old and to determine the association between occurrence of diarrhea, phylogenetic groups, and pathovars. Phylo-typing analysis of the 336 E. coli strains isolated from calves with Clermont method showed that 21 (6.25 %) belong to phylogroup A, 228 (67.85 %) to phylogroup B1, 2 (0.6 %) to phylogroup B2, 5 (1.49 %) to phylogroup C, 57 (16.96 %) to phylogroup E, and 3 (0.9 %) to phylogroup F. Phylogroup D was not identified and 20 strains (5.95 %) were assigned as "unknown." The distribution of phylogenetic groups among pathovars showed that NTEC belong to phylogroups B1 (17) and C (4); EPEC to phylogroups B1 (6) and E (8); STEC to phylogroups A (5), B1 (56), B2 (2), C (1), and E (15); EHEC to phylogroups B1 (95) and E (5); and ETEC to phylogroups A (3), B1 (7), and E (10). The EAST-1 strains were phylogroups A (13), B1 (47), E (19), and F (3); E. coli strains of "unknown" phylogroups belonged to pathovars EPEC (1), EHEC (2), STEC (7), and EAST-1 strains (6). ETEC was associated with diarrhea (P = 0.002). Our study did not find association between the phylogenetic background and occurrence of diarrhea (P = 0.164) but did find some relationship in phylogenetic group and pathovar. The study showed that EHEC and STEC are classified as phylogroup B1, EAST-1 phylogroup A, ETEC, and EPEC phylogroup E.
Asunto(s)
Enfermedades de los Bovinos/diagnóstico , Diarrea/veterinaria , Escherichia coli Enteropatógena/patogenicidad , Escherichia coli Enterotoxigénica/patogenicidad , Infecciones por Escherichia coli/veterinaria , Escherichia coli Shiga-Toxigénica/patogenicidad , Factores de Virulencia/genética , Animales , Técnicas de Tipificación Bacteriana , Brasil , Bovinos , Enfermedades de los Bovinos/microbiología , Enfermedades de los Bovinos/patología , ADN Bacteriano/genética , Diarrea/diagnóstico , Diarrea/microbiología , Diarrea/patología , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/crecimiento & desarrollo , Escherichia coli Enterotoxigénica/clasificación , Escherichia coli Enterotoxigénica/genética , Escherichia coli Enterotoxigénica/crecimiento & desarrollo , Infecciones por Escherichia coli/diagnóstico , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/patología , Heces/microbiología , Expresión Génica , Genotipo , Masculino , Filogenia , Reacción en Cadena de la Polimerasa , Escherichia coli Shiga-Toxigénica/clasificación , Escherichia coli Shiga-Toxigénica/genética , Escherichia coli Shiga-Toxigénica/crecimiento & desarrollo , Virulencia , Factores de Virulencia/metabolismoRESUMEN
Acute diarrheal disease (ADD) is a global public health problem, especially in developing countries and is one of the causes of mortality in children under five. ADD etiologic agents include viruses, bacteria and parasites in that order. Escherichia coli bacteria it is classified as a major diarrheagenic agent and transmitted by consuming contaminated water or undercooked foods. This review compiled updates on information virulence factors and pathogenic mechanisms involved in adhesion and colonization of seven pathotypes of E. coli called enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), shigatoxigenic E. coli (STEC), enteroaggregative E. coli (EAEC) and diffusely-adherent E. coli (DAEC). A final pathotype, adherent-invasive E. coli (AIEC) associated with Crohn's disease was also reviewed. The diarrheagenic pathotypes of E. coli affect different population groups and knowledge of the molecular mechanisms involved in the interaction with the human is important to guide research towards the development of vaccines and new tools for diagnosis and control.
Asunto(s)
Diarrea/microbiología , Escherichia coli Enteropatógena/patogenicidad , Infecciones por Escherichia coli/microbiología , Diarrea/virología , Escherichia coli Enteropatógena/clasificación , Heces/microbiología , Humanos , Factores de VirulenciaRESUMEN
Acute diarrheal disease (ADD) is a global public health problem, especially in developing countries and is one of the causes of mortality in children under five. ADD etiologic agents include viruses, bacteria and parasites in that order. Escherichia coli bacteria it is classified as a major diarrheagenic agent and transmitted by consuming contaminated water or undercooked foods. This review compiled updates on information virulence factors and pathogenic mechanisms involved in adhesion and colonization of seven pathotypes of E. coli called enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), shigatoxigenic E. coli (STEC), enteroaggregative E. coli (EAEC) and diffusely-adherent E. coli (DAEC). A final pathotype, adherent-invasive E. coli (AIEC) associated with Crohn's disease was also reviewed. The diarrheagenic pathotypes of E. coli affect different population groups and knowledge of the molecular mechanisms involved in the interaction with the human is important to guide research towards the development of vaccines and new tools for diagnosis and control.
La enfermedad diarreica aguda (EDA) es un problema de salud pública mundial, especialmente en los países en vía de desarrollo y es una de las causas de mortalidad en niños bajo cinco años de edad. Los agentes etiológicos de EDA incluyen virus, bacterias y parásitos, en ese orden. Dentro de las bacterias, Escherichia coli está clasificada como uno de los principales agentes diarreagénicos y se trasmite por el consumo de agua y alimentos contaminados o mal cocidos. Esta revisión recopiló información actualizada sobre los factores de virulencia y los mecanismos de patogenicidad implicados en la adhesión y colonización de siete patotipos de E. coli denominados, E. coli enteropatógena (ECEP), E. coli enterotoxigénica (ECET), E. coli enteroinvasora (ECEI), E. coli shigatoxigénica (ECST), E. coli enteroagregativa (ECEA) y E. coli de adherencia difusa (ECAD). Un último patotipo, E. coli adherente invasor (ECAI) asociado a la enfermedad de Crohn también fue revisado. Los patotipos diarreagénicos de E. coli afectan a diferentes grupos poblacionales y el conocimiento de los mecanismos moleculares implicados en la interacción con el humano es importante para orientar las investigaciones hacia el desarrollo de vacunas o nuevas herramientas para su diagnóstico y control.
Asunto(s)
Humanos , Diarrea/microbiología , Infecciones por Escherichia coli/microbiología , Escherichia coli Enteropatógena/patogenicidad , Factores de Virulencia , Diarrea/virología , Escherichia coli Enteropatógena/clasificación , Heces/microbiologíaRESUMEN
Enteropathogenic Escherichia coli (EPEC) are important agents of diarrhoea in industrialized as well as developing countries, such as Brazil. The hallmark of EPEC pathogenesis is the establishment of attaching and effacing lesions in enterocytes, in which pedestal-like structures are formed underneath adherent bacteria. EPEC are divided into two subgroups, typical (tEPEC) and atypical (aEPEC), based on the presence of the EPEC adherence factor plasmid in tEPEC and its absence in aEPEC. This study was designed to characterize 82 aEPEC isolates obtained from stool samples of diarrhoeic patients during 2012 and 2013 in Brazil. The majority of the aEPEC were assigned to the phylo-group B1 (48.8 %), and intimin subtypes θ (20.7 %), ß1 (9.7 %) and λ (9.7 %) were the most prevalent among the isolates. The nleB and nleE genes were concomitantly detected in 32.9 % of the isolates, demonstrating the occurrence of the pathogenicity island O122 among them. The O157-plasmid genes (ehxA and/or espP) were detected in 7.3 % of the isolates, suggesting that some aEPEC could be derived from Shiga-toxin-producing E. coli that lost the stx genes while trafficking in the host. PFGE of 14 aEPEC of serotypes O2 : H16, O33 : H34, O39 : H9, O108 : H- and ONT : H19 isolated from five distinct outbreaks showed serotype-specific PFGE clusters, indicating a high degree of similarity among the isolates from the same event, thus highlighting these serotypes as potential aetiologic agents of diarrhoeal outbreaks in Brazil.
Asunto(s)
Diarrea/epidemiología , Diarrea/microbiología , Brotes de Enfermedades , Escherichia coli Enteropatógena/aislamiento & purificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Variación Genética , Adhesinas Bacterianas/genética , Adulto , Anciano de 80 o más Años , Brasil/epidemiología , Niño , Preescolar , Electroforesis en Gel de Campo Pulsado , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/genética , Proteínas de Escherichia coli/genética , Heces/microbiología , Femenino , Genes Bacterianos , Genotipo , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Tipificación Molecular , Plásmidos/análisis , Serogrupo , Adulto JovenRESUMEN
The aim of the present study was to investigate the prevalence of Diarrheagenic Escherichia coli (DEC) pathotypes, a leading cause of diarrhea worldwide, among diarrheal and healthy children, up to 5 years of age, living in the city of Botucatu, São Paulo, Brazil. DEC, investigated by PCR detection of virulence factor-encoding genes associated with the distinct pathotypes, was isolated from 18.0% of the patients, and 19.0% of the controls, with enteroaggregative E. coli (EAEC), the most frequent pathotype, being detected in equal proportion between patients and controls (10.0%). Among the enteropathogenic E. coli (EPEC) isolates, only one isolate was able to produce the localized adherence pattern to HeLa cells, being thus the only typical EPEC identified. All the remaining EPEC were classified as atypical (aEPEC), and detected in 8.0% and 8.5% of the patients and controls, respectively. Regarding the serotypes, 26.5% of the analyzed EPEC isolates belonged to classical EPEC-serogroups, and the only two STEC found were serotyped as O26:H11 (patient) and O119:H7 (control). Antimicrobial susceptibility tests revealed that 43.6%, 29.5% and 2.6% of the DEC isolates were resistant to ampicillin, cotrimoxazole and gentamicin, respectively. Our data indicate that EAEC remains prevalent among children living in Botucatu, and revealed atypical EPEC as emerging putative diarrheal agents in this geographical region.
Asunto(s)
Diarrea/diagnóstico , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/patogenicidad , Infecciones por Escherichia coli/diagnóstico , Factores de Virulencia/genética , Ampicilina/farmacología , Antibacterianos/farmacología , Adhesión Bacteriana , Técnicas de Tipificación Bacteriana , Brasil/epidemiología , Preescolar , Diarrea/tratamiento farmacológico , Diarrea/epidemiología , Diarrea/microbiología , Farmacorresistencia Bacteriana , Escherichia coli Enteropatógena/genética , Escherichia coli Enteropatógena/aislamiento & purificación , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/microbiología , Heces/microbiología , Femenino , Genotipo , Gentamicinas/farmacología , Células HeLa , Humanos , Lactante , Recién Nacido , Masculino , Pruebas de Sensibilidad Microbiana , Prevalencia , Serogrupo , Combinación Trimetoprim y Sulfametoxazol/farmacologíaRESUMEN
Escherichia coli enteropatógeno (EPEC) es uno de los principales agentes de diarrea infantil aguda en los países en desarrollo. Se clasifica en típico (tEPEC) y atípico (aEPEC) sobre la base de la presencia del factor bfp, asociado a la adherencia y codificado en el plásmido pEAF. Se describe el aislamiento de E. coli O157:H16, de la categoría aEPEC, en un caso de diarrea sanguinolenta infantil y en sus contactos familiares. De las muestras de materia fecal del niño, de la madre, del padre y de la hermana se aisló E. coli O157:H16 eae-??-positivo, sorbitol-positivo, ß-glucuronidasa-positivo, sensible a los antimicrobianos ensayados, y negativo para los factores stx1, stx2, ehxA y bfp. Por XbaI-PFGE, todos los aislamientos presentaron el patrón de macrorrestricción AREXHX01.1040, con 100% de similitud. Es importante la vigilancia epidemiológica de los casos de diarrea asociados a E. coli O157 y sus contactos familiares, y la incorporación de técnicas para detectar los distintos patotipos de E. coli
Enteropathogenic Escherichia coli (EPEC) is a major causative agent of acute diarrhea in children in developing countries. This pathotype is divided into typical EPEC (tEPEC) and atypical EPEC (aEPEC), based on the presence of the bfp virulence factor associated with adhesion, encoded in the pEAF plasmid. In the present study, the isolation of aEPEC O157:H16 from a bloody diarrhea case in a child and his household contacts (mother, father and sister) is described. The strain was characterized as E. coli O157:H16 eae-??-positive, sorbitol fermenter with ß-glucuronidase activity, susceptible to all antimicrobials tested, and negative for virulence factors stx1, stx2, ehxA and bfp. XbaI-PFGE performed on all isolates showed the AREXHX01.1040 macrorestriction pattern, with 100% similarity. These results highlight the importance of epidemiological surveillance of E. coli O157-associated diarrhea cases identified in children and their family contacts, as well as the incorporation of molecular techniques that allow the detection of the different E. coli pathotypes
Asunto(s)
Humanos , Masculino , Preescolar , Diarrea Infantil/prevención & control , Escherichia coli Enteropatógena/clasificación , Familia , Escherichia coli O157/aislamiento & purificación , Monitoreo Epidemiológico , AntiinfecciososRESUMEN
The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli . Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli . Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpA gene . EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg , aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea ( P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.
Asunto(s)
Diarrea/microbiología , Escherichia coli Enteropatógena/genética , Infecciones por Escherichia coli/microbiología , Escherichia coli/genética , Factores de Virulencia/genética , Adhesión Bacteriana , Brasil , Niño , Preescolar , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Enteropatógena/fisiología , Células Epiteliales/microbiología , Escherichia coli/clasificación , Escherichia coli/aislamiento & purificación , Escherichia coli/fisiología , Humanos , Lactante , Recién Nacido , Antígenos O/análisis , SerogrupoRESUMEN
The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli. Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli. Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpA gene. EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg, aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea (P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.
Asunto(s)
Niño , Preescolar , Humanos , Lactante , Recién Nacido , Diarrea/microbiología , Escherichia coli Enteropatógena/genética , Infecciones por Escherichia coli/microbiología , Escherichia coli/genética , Factores de Virulencia/genética , Adhesión Bacteriana , Brasil , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/aislamiento & purificación , Escherichia coli Enteropatógena/fisiología , Células Epiteliales/microbiología , Escherichia coli/clasificación , Escherichia coli/aislamiento & purificación , Escherichia coli/fisiología , Antígenos O/análisis , SerogrupoRESUMEN
The bacterial pathogen most commonly associated with endemic forms of childhood diarrhoea is Escherichia coli. Studies of epidemiological characteristics of HEp-2 cell-adherent E. coli in diarrhoeal disease are required, particularly in developing countries. The aim of this study was evaluate the presence and significance of adherent Escherichia coli from diarrhoeal disease in children. The prevalence of LA, AA, and DA adherence patterns were determined in HEp-2 cells, the presence of virulence genes and the presence of the O serogroups in samples obtained from 470 children with acute diarrhoea and 407 controls in Porto Velho, Rondônia, Brazil. E. coli isolates were identified by PCR specific for groups of adherent E. coli. Out of 1,156 isolates obtained, 128 (11.0%) were positive for eae genes corresponding to EPEC, however only 38 (29.6%) of these amplified bfpA gene. EAEC were isolated from 164 (14.1%) samples; of those 41(25%), 32 (19%) and 16 (9.7%) amplified eagg, aggA or aafA genes, respectively and aggA was significantly associated with diarrhoea (P = 0.00006). DAEC identified by their adhesion pattern and there were few isolates. In conclusion, EAEC was the main cause of diarrhoea in children, especially when the aggA gene was present, followed by EPEC and with a negligible presence of DAEC.(AU)
Asunto(s)
Humanos , Recién Nacido , Lactante , Preescolar , Niño , Diarrea/microbiología , Escherichia coli Enteropatógena/genética , Infecciones por Escherichia coli/microbiología , Escherichia coli/genética , Factores de Virulencia/genética , Adhesión Bacteriana , Brasil , Escherichia coli Enteropatógena/clasificación , Escherichia coli Enteropatógena/aislamiento & purificaciónRESUMEN
The aim of this study was to investigate the prevalence of Shiga toxin-producing Escherichia coli (STEC) and enteropathogenic E. coli (EPEC) strains and to identify the stx gene types in wild captive and companion birds. In total,657 E. coli isolates from 219 birds belonging to 38 different species were investigated for the presence of STEC and EPEC strains. It was shown that five birds (2.28%) carried strains positive for one or more of the virulence factors investigated. The results indicated that 1.8% (n=4) and 0.45% (n=1) of the birds carried STEC and EPEC strains, respectively. All STEC strains harbored the stx2f and eae genes and this finding reveals the role of other birds, in addition to pigeons, as reservoirs of STEC. The only EPEC strain in this study was isolated from a Myna. Based on our knowledge, this is the first report of Stx2f-producing STEC in Geese, Duck and Lesser kestrel. In conclusion, the results indicate a low frequency of STEC carriage in wild and companion birds, and point out the need of additionally screening for the presence of stx2f in all the eae-harboring strains from birds.(AU)