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Gastroenteritis/diagnóstico , Técnicas de Diagnóstico Molecular/estadística & datos numéricos , Técnicas de Diagnóstico Molecular/normas , Toma de Decisiones Clínicas , Pruebas Diagnósticas de Rutina/normas , Pruebas Diagnósticas de Rutina/estadística & datos numéricos , Reacciones Falso Positivas , Gastroenteritis/terapia , Humanos , Valor Predictivo de las PruebasRESUMEN
OBJECTIVES: To evaluate the relationship between individual bacterial and viral pathogens and disease severity. METHODS: Children <18 years with three or more episodes of vomiting and/or diarrhoea were enrolled in two Canadian paediatric emergency departments between December 2014 and August 2016. Specimens were analysed employing molecular panels, and outcome data were collected 14 days after enrolment. The primary outcome was severe disease over the entire illness (symptom onset until 14-day follow-up), quantified employing the Modified Vesikari Scale (MVS) score. The score was additionally analysed in two other time periods: index (symptom onset until enrolment) and follow-up (enrolment until 14-day follow-up). RESULTS: Median participant age was 20.7 (IQR: 11.3, 44.2) months; 47.4% (518/1093) and 73.4% (802/1093) of participants had index and total MVS scores ≥11, respectively. The most commonly identified pathogens were rotavirus (289/1093; 26.4%) and norovirus (258/1093; 23.6%). In multivariable analysis, severe disease over the entire illness was associated with rotavirus (OR = 9.60; 95%CI: 5.69, 16.19), Salmonella (OR = 6.61; 95%CI: 1.50, 29.17), adenovirus (OR = 2.53; 95%CI: 1.62, 3.97), and norovirus (OR = 1.43; 95%CI: 1.01, 2.01). Pathogens associated with severe disease at the index visit were: rotavirus only (OR = 6.13; 95%CI: 4.29, 8.75), Salmonella (OR = 4.59; 95%CI: 1.71, 12.29), adenovirus only (OR = 2.06; 95%CI: 1.41, 3.00), rotavirus plus adenovirus (OR = 3.15; 95%CI: 1.35, 7.37), and norovirus (OR = 0.68; 95%CI: 0.49, 0.94). During the follow-up period, rotavirus (OR = 2.21; 95%CI: 1.50, 3.25) and adenovirus (OR = 2.10; 95%CI: 1.39, 3.18) were associated with severe disease. CONCLUSIONS: In children presenting for emergency department care with acute gastroenteritis, pathogens identified were predominantly viruses, and several of which were associated with severe disease. Salmonella was the sole bacterium independently associated with severe disease.
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Adenoviridae/aislamiento & purificación , Gastroenteritis , Norovirus/aislamiento & purificación , Rotavirus/aislamiento & purificación , Salmonella/aislamiento & purificación , Adolescente , Adulto , Canadá , Niño , Gastroenteritis/diagnóstico , Gastroenteritis/tratamiento farmacológico , Gastroenteritis/microbiología , Humanos , Lactante , Estudios Prospectivos , Resultado del Tratamiento , Adulto JovenRESUMEN
Necrotizing enterocolitis (NEC) remains a major challenge in neonatology. Little is known about NEC pathophysiology apart from the presence of pre-event gut dysbiosis. Here, we applied broad range metabolomics to stools obtained 1-5 days before NEC developed from 9 cases (9 samples) and 19 (32 samples) controls matched for gestational age at birth and birth weight. The 764 identified metabolites identified six pathways that differ between cases and controls. We pursued sphingolipid metabolism because cases had decreased ceramides and increased sphingomyelins compared to controls, and because of the relevance of sphingolipids to human inflammatory disorders. Targeted analysis of samples from 23 cases and 46 controls confirmed the initial broad range observations. While metabolites provided only 73% accuracy of classification by machine learning, hierarchical clustering defined a sphingolipid associated grouping that contained 60% of the cases but only 13% of the controls, possibly identifying a pathophysiologically distinct subset of NEC. The clustering did not associate with any of the analyzed clinical and sample variables. We conclude that there are significant changes in sphingolipid metabolism components in pre-NEC stools compared to controls, but our data urge circumspection before using sphingolipids as broadly applicable predictive biomarkers.
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Enterocolitis Necrotizante/etiología , Contenido Digestivo/química , Esfingolípidos/análisis , Biomarcadores/análisis , Peso al Nacer , Estudios de Casos y Controles , Enterocolitis Necrotizante/diagnóstico , Heces/química , Femenino , Edad Gestacional , Humanos , Recién Nacido , Masculino , Metabolómica/métodosRESUMEN
Escherichia coli O157:H7 is the largest cause of hemolytic uremic syndrome (HUS). Previous studies proposed that HUS risk varies across the E. coli O157:H7 phylogenetic tree (hypervirulent clade 8), but the role of age in the association is unknown. We determined phylogenetic lineage of E. coli O157:H7 isolates from 1160 culture-confirmed E. coli O157:H7 cases reported in Washington State, 2004-2015. Using generalised estimating equations, we tested the association between phylogenetic lineage and HUS. Age was evaluated as an effect modifier. Among 1082 E. coli O157:H7 cases with both phylogenetic lineage and HUS status (HUS n = 76), stratified analysis suggested effect modification by age. Lineages IIa and IIb, relative to Ib, did not appear associated with HUS in children 0-9-years-old. For cases 10-59-years-old, lineages IIa and IIb appeared to confer increased risk of HUS, relative to lineage Ib. The association reversed in ⩾60-year-olds. Results were similar for clade 8. Phylogenetic lineage appears to be associated with HUS risk only among those ⩾10-years-old. Among children <10, the age group most frequently affected, lineage does not explain progression to HUS. However, lineage frequency varied across age groups, suggesting differences in exposure and/or early disease manifestation.
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Infecciones por Escherichia coli/microbiología , Escherichia coli O157/aislamiento & purificación , Síndrome Hemolítico-Urémico/microbiología , Adolescente , Adulto , Factores de Edad , Niño , Preescolar , Infecciones por Escherichia coli/epidemiología , Femenino , Síndrome Hemolítico-Urémico/epidemiología , Humanos , Lactante , Recién Nacido , Masculino , Persona de Mediana Edad , Filogenia , Estudios Retrospectivos , Washingtón/epidemiologíaRESUMEN
PURPOSE: The enteric microbiome is known to play a major role in healthy gut homeostasis and several disease states. It may also contribute to both the intestinal recovery and complications that occur in patients with short bowel syndrome. The extent and nature of alterations to the gut microbiota following intestinal resection, however, are not well studied in a controlled setting. The purpose of this investigation is to characterize the effects of massive small bowel resection on the murine enteric microflora. METHODS: Wild-type C57BL6 mice, following a week of acclamation to a liquid rodent diet, underwent either 50% proximal small bowel resection (SBR) or a sham operation. Mice were sacrificed, and enteric contents from the small bowel, cecum, and stool were harvested at 7 and 90 days post-operatively. DNA was isolated, and the V3-V5 regions of the 16s rRNA gene amplified and pyrosequenced on a Roche 454 platform. Sequences were clustered into operation taxonomic units and classified. Communities were then analyzed for diversity and phylogenic composition. RESULTS: In the long-term group, the microbes inhabiting the ileum of mice undergoing SBR and sham operation differed significantly at the genus level (p < 0.001). Small bowel contents collected before and after SBR also differed significantly (p = 0.006). This was driven by an increase in Lactobacillus and decrease in Enterobacteriaceae species in mice undergoing SBR. No difference was seen in the long-term stool or in stool, cecal, or ileal contents in the short-term. No difference in microbial community diversity was found in any group. CONCLUSION: Bowel resection induces long-term changes in the microbial community of the murine ileum, but not at more distal sites of the gastrointestinal tract. The increase in Lactobacillus encountered small bowel of resected mice correlates with limited previous studies. These changes may reflect an adaptive response of the microbiota to maximize energy extraction, but further studies are needed to establish the role played by this altered community.
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Bacterias/aislamiento & purificación , Mucosa Intestinal/microbiología , Intestino Delgado/cirugía , Microbiota/fisiología , Síndrome del Intestino Corto/microbiología , Animales , Modelos Animales de Enfermedad , Estudios de Seguimiento , Mucosa Intestinal/cirugía , Intestino Delgado/microbiología , Ratones , Ratones Endogámicos C57BLRESUMEN
OBJECTIVE: Model age of necrotizing enterocolitis (NEC) onset applying Sartwell's model of incubation periods, and examine its relationship to gestational age (GA). STUDY DESIGN: Retrospective chart review of St Louis Children's Hospital neonates diagnosed with NEC (≥Bell's stage II) from 2004 to 2008, inclusive. RESULT: The relationship between age of NEC (N=84 cases) onset and GA best fits a non-linear model, with infants ≤28 weeks having a disproportionately longer time to onset than older GA groups and explained 50.3% of the variability in age of NEC onset. Additional clinical variables provided no improvement in explaining age of NEC onset. Application of Sartwell's model to age of NEC onset proved a good fit, when birth is used as the common exposure episode, and age is equivalent of the incubation period. CONCLUSION: The relationship between day of NEC diagnosis and GA is non-linear, with lower GA infants having disproportionately longer time to onset. Despite these GA differences, the fit to Sartwell's model for incubation periods model is consistent with NEC being a consequence of an event that occurs at or soon after birth.
Asunto(s)
Enterocolitis Necrotizante/epidemiología , Edad Gestacional , Periodo de Incubación de Enfermedades Infecciosas , Edad de Inicio , Femenino , Humanos , Recién Nacido , Masculino , Modelos Estadísticos , Dinámicas no LinealesRESUMEN
Clostridium difficile is a well recognized pathogen of humans and animals. Although C. difficile was first identified over 70 years ago, much remains unknown in regards to the primary source of human acquisition and its pathobiology. These deficits in our knowledge have been intensified by dramatic increases in both the frequency and severity of disease in humans over the last decade. The changes in C. difficile epidemiology might be due to the emergence of a hypervirulent stain of C. difficile, ageing of the population, altered risk of developing infection with newer medications, and/or increased exposure to C. difficile outside of hospitals. In recent years, there have been numerous reports documenting C. difficile contamination of various foods, and reports of similarities between strains that infect animals and strains that infect humans as well. The purposes of this review are to highlight the many challenges to diagnosing, treating, and preventing C. difficile infection in humans, and to stress that collaboration between human and veterinary researchers is needed to control this pathogen.
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Clostridioides difficile/patogenicidad , Enterocolitis Seudomembranosa/transmisión , Enterocolitis Seudomembranosa/veterinaria , Zoonosis , Animales , Enfermedades Transmisibles Emergentes/epidemiología , Enfermedades Transmisibles Emergentes/prevención & control , Enfermedades Transmisibles Emergentes/transmisión , Enfermedades Transmisibles Emergentes/veterinaria , Enterocolitis Seudomembranosa/epidemiología , Enterocolitis Seudomembranosa/prevención & control , Contaminación de Alimentos , Microbiología de Alimentos , Humanos , Incidencia , Control de Infecciones/métodos , Factores de Riesgo , VirulenciaAsunto(s)
Diarrea/epidemiología , Diarrea/mortalidad , Micronutrientes/deficiencia , Preescolar , Enfermedad Crónica , Trastornos del Conocimiento/epidemiología , Trastornos del Conocimiento/etiología , Deshidratación/prevención & control , Diarrea/prevención & control , Trastornos del Crecimiento/epidemiología , Trastornos del Crecimiento/etiología , Humanos , Lactante , Recién Nacido , Micronutrientes/uso terapéutico , Investigación , Zinc/uso terapéuticoRESUMEN
A survey for Shiga toxigenic Escherichia coli in raw milk and beef was conducted within a defined geographic region of the United States. Prevalence rates based on detection of Shiga toxin gene (stx) were 36% for retail beef, 23% for beef carcasses, and 21% for raw milk samples, which were significantly higher than were Shiga toxigenic E. coli isolation rates of 7.5, 5.8, and 3.2%, respectively. Seasonal prevalence differences were significant for stx positivity among ground beef and milk samples. Distribution of stx subtypes among isolates varied according to sample type, with stx1 predominating in milk, stx2 on carcasses, and the combination of both stx1 and stx2 in beef. Ancillary virulence markers eae and ehx were evident in 23 and 15% of isolates, respectively. Pulsed-field gel electrophoresis demonstrated associations between food isolates and sympatric bovine fecal, and human clinical isolates. These data demonstrate that non-O157 Shiga toxigenic E. coli is present in the food chain in the Pacific Northwest, and its risk to health warrants critical assessment.
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Contaminación de Alimentos/análisis , Carne/microbiología , Leche/microbiología , Toxina Shiga/análisis , Escherichia coli Shiga-Toxigénica/aislamiento & purificación , Animales , Bovinos , Seguridad de Productos para el Consumidor , Electroforesis en Gel de Campo Pulsado , Humanos , Productos de la Carne/microbiología , Prevalencia , Estaciones del Año , Estados Unidos , VirulenciaRESUMEN
E. coli O157:H7 can cause potentially lethal illness in hosts of all ages. These patients often are evaluated and treated by gastroenterologists. The treating physician should administer adequate hydration, usually parenterally, and avoid the use of antibiotics and antimotility agents. The physician needs to notify immediately the appropriate public health authorities of the diagnosis and to ensure that the isolate is recovered by the microbiologist and forwarded for molecular linkage analyses.
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Infecciones por Escherichia coli/diagnóstico , Escherichia coli O157/aislamiento & purificación , Anciano , Niño , Preescolar , Diagnóstico Diferencial , Infecciones por Escherichia coli/terapia , Escherichia coli O157/patogenicidad , Fluidoterapia/métodos , Humanos , Lactante , Toxinas Shiga/aislamiento & purificación , VirulenciaRESUMEN
The selC tRNA gene is a common site for the insertion of pathogenicity islands in a variety of bacterial enteric pathogens. We demonstrate here that Escherichia coli that produces Shiga toxin 2d and does not harbor the locus of enterocyte effacement (LEE) contains, instead, a novel genomic island. In one representative strain (E. coli O91:H(-) strain 4797/97), this island is 33,014 bp long and, like LEE in E. coli O157:H7, is integrated 15 bp downstream of selC. This E. coli O91:H(-) island contains genes encoding a novel serine protease, termed EspI; an adherence-associated locus, similar to iha of E. coli O157:H7; an E. coli vitamin B12 receptor (BtuB); an AraC-type regulatory module; and four homologues of E. coli phosphotransferase proteins. The remaining sequence consists largely of complete and incomplete insertion sequences, prophage sequences, and an intact phage integrase gene that is located directly downstream of the chromosomal selC. Recombinant EspI demonstrates serine protease activity using pepsin A and human apolipoprotein A-I as substrates. We also detected Iha-reactive protein in outer membranes of a recombinant clone and 10 LEE-negative, Shiga toxin-producing E. coli (STEC) strains by immunoblot analysis. Using PCR analysis of various STEC, enteropathogenic E. coli, enterotoxigenic E. coli, enteroaggregative E. coli, uropathogenic E. coli, and enteroinvasive E. coli strains, we detected the iha homologue in 59 (62%) of 95 strains tested. In contrast, espI and btuB were present in only two (2%) and none of these strains, respectively. We conclude that the newly described island occurs exclusively in a subgroup of STEC strains that are eae negative and contain the variant stx(2d )gene.
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Escherichia coli/enzimología , Serina Endopeptidasas/genética , Secuencia de Aminoácidos , Proteínas de la Membrana Bacteriana Externa , Proteínas Bacterianas/genética , Sitios de Unión , Enterocitos , Escherichia coli/genética , Escherichia coli/aislamiento & purificación , Escherichia coli/patogenicidad , Proteínas de Escherichia coli/genética , Expresión Génica , Genoma Bacteriano , Humanos , Proteínas de Transporte de Membrana , Datos de Secuencia Molecular , Mutagénesis Insercional , Sistemas de Lectura Abierta , Receptores de Péptidos/genética , Análisis de Secuencia de ADN , Serina Endopeptidasas/metabolismo , Toxina Shiga , VirulenciaRESUMEN
Hemolytic uremic syndrome (HUS) of childhood most commonly follows gastrointestinal infection with Escherichia coli O157:H7. This pathogen elaborates Shiga toxins that are believed to cause microvascular injury and to trigger a thrombogenic response. The exact mechanisms leading to variable disease manifestations are unknown. Allelic variation in genes encoding selected coagulation factors and inhibitors of fibrinolysis were examined to determine whether or not a causal relationship exists between hypercoagulability and the development of HUS. No correlation between the thrombogenic factor V (G1691A), factor II (G20210A), methylenetetrahydrofolate reductase (C677T), or the plasminogen activator inhibitor (PAI)-1 promotor (4G/5G) genotypes and the risk of infection with E. coli O157:H7, or the risk of development of HUS among infected patients, was found. Serum PAI-1 levels did not correlate with the PAI-1 genotype. We conclude that the alleles studied are not major risk factors for the acquisition of E. coli O157:H7 infection, or of E. coli O157:H7-related HUS.
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Coagulación Sanguínea/genética , Infecciones por Escherichia coli/genética , Síndrome Hemolítico-Urémico/genética , Alelos , Factor V/genética , Humanos , Metilenotetrahidrofolato Reductasa (NADPH2) , Oxidorreductasas actuantes sobre Donantes de Grupo CH-NH/genética , Inhibidor 1 de Activador Plasminogénico/genética , Protrombina/genéticaRESUMEN
Hemolytic uremic syndrome (HUS) usually occurs after infection with Shiga toxin-producing bacteria. Thrombotic thrombocytopenic purpura, a disorder with similar clinical manifestations, is associated with deficient activity of a circulating metalloprotease that cleaves von Willebrand factor at the Tyr842-Met843 peptide bond in a shear stress-dependent manner. We analyzed von Willebrand factor-cleaving metalloprotease activity and the status of von Willebrand factor in 16 children who developed HUS after Escherichia coli O157:H7 infection and in 29 infected children who did not develop this complication. Von Willebrand factor-cleaving metalloprotease activity was normal in all subjects, but von Willebrand factor size was decreased in the plasma of each of 16 patients with HUS. The decrease in circulating von Willebrand factor size correlated with the severity of thrombocytopenia and was proportional to an increase in von Willebrand factor proteolytic fragments in plasma. Immunohistochemical studies of the kidneys in four additional patients who died of HUS demonstrated glomerular thrombi in three patients, and arterial and arteriolar thrombi in one patient. The glomerular thrombi contained fibrin but little or no von Willebrand factor. A decrease in large von Willebrand factor multimers, presumably caused by enhanced proteolysis from abnormal shear stress in the microcirculation, is common in HUS.
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Infecciones por Escherichia coli/sangre , Escherichia coli O157/aislamiento & purificación , Síndrome Hemolítico-Urémico/sangre , Metaloendopeptidasas/sangre , Factor de von Willebrand/metabolismo , Proteínas ADAM , Proteína ADAMTS13 , Niño , Preescolar , Infecciones por Escherichia coli/enzimología , Femenino , Síndrome Hemolítico-Urémico/enzimología , Síndrome Hemolítico-Urémico/microbiología , Humanos , Hidrólisis , Inmunohistoquímica , Riñón/patología , MasculinoRESUMEN
The rfb region specifies the structure of lipopolysaccharide side chains that comprise the diverse gram-negative bacterial somatic (O) antigens. The rfb locus is adjacent to gnd, which is a polymorphic gene encoding 6-phosphogluconate dehydrogenase. To determine if rfb and gnd cotransfer, we sequenced gnd in five O55 and 13 O157 strains of Escherichia coli. E. coli O157:H7 has a gnd allele (allele A) that is only 82% identical to the gnd allele (allele D) of closely related E. coli O55:H7. In contrast, gnd alleles of E. coli O55 in distant lineages are >99.9% identical to gnd allele D. Though gnd alleles B and C in E. coli O157 that are distantly related to E. coli O157:H7 are more similar to allele A than to allele D, there are nucleotide differences at 4 to 6% of their sites. Alleles B and C can be found in E. coli O157 in different lineages, but we have found allele A only in E. coli O157 belonging to the DEC5 lineage. DNA 3' to the O55 gnd allele in diverse E. coli lineages has sequences homologous to tnpA of the Salmonella enterica serovar Typhimurium IS200 element, E. coli Rhs elements (including an H-rpt gene), and portions of the O111 and O157 rfb regions. We conclude that rfb and gnd cotransferred into E. coli O55 and O157 in widely separated lineages and that recombination was responsible for recent antigenic shifts in the emergence of pathogenic E. coli O55 and O157.
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Proteínas Bacterianas/genética , Escherichia coli/genética , Genoma Bacteriano , Antígenos O/genética , Fosfogluconato Deshidrogenasa/genética , Alelos , Secuencia de Bases , Evolución Biológica , Clonación Molecular , Escherichia coli/fisiología , Escherichia coli O157/genética , Escherichia coli O157/fisiología , Recombinación Genética , Homología de Secuencia de Ácido Nucleico , Factores de Transcripción/genéticaRESUMEN
We evaluated the Meridian IC-STAT direct fecal and broth culture antigen detection methods with samples from children infected with Escherichia coli O157:H7 and correlated the antigen detection results with the culture results. Stools of 16 children who had recently had stool cultures positive for this pathogen (population A) and 102 children with diarrhea of unknown cause (population B) were tested with the IC-STAT device (direct testing). Fecal broth cultures were also tested with this device (broth testing). The results were correlated to a standard of the combined yield from direct culture of stools on sorbitol-MacConkey (SMAC) agar and culture of broth on SMAC agar. Eleven (69%) of the population A stool specimens yielded E. coli O157:H7 when plated directly on SMAC agar. Two more specimens yielded this pathogen when the broth culture was similarly plated. Of these 13 stool specimens, 8 and 13 were positive by direct and broth testing (respective sensitivities, 62 and 100%). Compared to the sensitivity of a simultaneously performed SMAC agar culture, the sensitivity of direct testing was 73%. Three (3%) of the population B stool specimens contained E. coli O157:H7 on SMAC agar culture; one and three of these stool specimens were positive by direct and broth testing, respectively. The direct and broth IC-STAT tests were 100% specific with samples from children from population B. Direct IC-STAT testing of stools is rapid, easily performed, and specific but is insufficiently sensitive to exclude the possibility of infection with E. coli O157:H7. Performing the IC-STAT test with a broth culture increases its sensitivity. However, attempts to recover E. coli O157:H7 by culture should not be abandoned but, rather, should be increased when the IC-STAT test result is positive.
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Antígenos Bacterianos/análisis , Infecciones por Escherichia coli/microbiología , Escherichia coli O157/aislamiento & purificación , Heces/microbiología , Niño , Preescolar , Cromatografía , Medios de Cultivo , Diarrea/microbiología , Escherichia coli O157/clasificación , Escherichia coli O157/crecimiento & desarrollo , Escherichia coli O157/inmunología , Humanos , InmunoensayoRESUMEN
BACKGROUND: Children with gastrointestinal infections caused by Escherichia coli O157:H7 are at risk for the hemolytic-uremic syndrome. Whether antibiotics alter this risk is unknown. METHODS: We conducted a prospective cohort study of 71 children younger than 10 years of age who had diarrhea caused by E. coli O157:H7 to assess whether antibiotic treatment in these children affects the risk of the hemolytic-uremic syndrome and to assess the influence of confounding factors on this outcome. Estimates of relative risks were adjusted for possible confounding effects with the use of logistic-regression analysis. RESULTS: Among the 71 children, 9 (13 percent) received antibiotics and the hemolytic-uremic syndrome developed in 10 (14 percent). Five of these 10 children had received antibiotics. Factors significantly associated with the hemolytic-uremic syndrome were a higher initial white-cell count (relative risk, 1.3; 95 percent confidence interval, 1.1 to 1.5), evaluation with stool culture soon after the onset of illness (relative risk, 0.3; 95 percent confidence interval, 0.2 to 0.8), and treatment with antibiotics (relative risk, 14.3; 95 percent confidence interval, 2.9 to 70.7). The clinical and laboratory characteristics of the 9 children who received antibiotics and the 62 who did not receive antibiotics were similar. In a multivariate analysis that was adjusted for the initial white-cell count and the day of illness on which stool was obtained for culture, antibiotic administration remained a risk factor for the development of the hemolytic uremic syndrome (relative risk, 17.3; 95 percent confidence interval, 2.2 to 137). CONCLUSIONS: Antibiotic treatment of children with E. coli O157:H7 infection increases the risk of the hemolytic-uremic syndrome.
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Antibacterianos/efectos adversos , Diarrea/tratamiento farmacológico , Infecciones por Escherichia coli/tratamiento farmacológico , Escherichia coli O157 , Síndrome Hemolítico-Urémico/etiología , Antibacterianos/uso terapéutico , Cefalosporinas/efectos adversos , Niño , Preescolar , Diarrea/complicaciones , Diarrea/microbiología , Infecciones por Escherichia coli/complicaciones , Infecciones por Escherichia coli/microbiología , Heces/microbiología , Femenino , Humanos , Lactante , Modelos Logísticos , Masculino , Estudios Prospectivos , Riesgo , Combinación Trimetoprim y Sulfametoxazol/efectos adversosRESUMEN
Two novel putative Escherichia coli virulence genes, iha and iroN from E. coli (iroN(E. coli)), were detected in 55 and 39%, respectively, of 67 E. coli isolates from patients with urosepsis. iha and iroN(E. coli) exhibited divergent associations with other putative virulence genes, phylogenetic markers, host characteristics, and antimicrobial resistance.
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Proteínas Bacterianas/genética , Proteínas de Escherichia coli , Escherichia coli/genética , Genes Bacterianos , Sepsis/microbiología , Adulto , Alelos , Proteínas Bacterianas/clasificación , Infecciones por Escherichia coli/epidemiología , Infecciones por Escherichia coli/inmunología , Proteínas Fimbrias , Humanos , Filogenia , VirulenciaRESUMEN
The mechanisms used by Shiga toxin (Stx)-producing Escherichia coli to adhere to epithelial cells are incompletely understood. Two cosmids from an E. coli O157:H7 DNA library contain an adherence-conferring chromosomal gene encoding a protein similar to iron-regulated gene A (IrgA) of Vibrio cholerae (M. B. Goldberg, S. A. Boyko, J. R. Butterton, J. A. Stoebner, S. M. Payne, and S. B. Calderwood, Mol. Microbiol. 6:2407-2418, 1992). We have termed the product of this gene the IrgA homologue adhesin (Iha), which is encoded by iha. Iha is 67 kDa in E. coli O157:H7 and 78 kDa in laboratory E. coli and is structurally unlike other known adhesins. DNA adjacent to iha contains tellurite resistance loci and is conserved in structure in distantly related pathogenic E. coli, but it is absent from nontoxigenic E. coli O55:H7, sorbitol-fermenting Stx-producing E. coli O157:H-, and laboratory E. coli. We have termed this region the tellurite resistance- and adherence-conferring island. We conclude that Iha is a novel bacterial adherence-conferring protein and is contained within an E. coli chromosomal island of conserved structure. Pathogenic E. coli O157:H7 has only recently acquired this island.
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Adhesinas Bacterianas/genética , Adhesión Bacteriana , Cromosomas Bacterianos , Escherichia coli O157/genética , Secuencia de Aminoácidos , Escherichia coli O157/clasificación , Escherichia coli O157/fisiología , Datos de Secuencia MolecularRESUMEN
OBJECTIVE: Numerous investigators have proposed a role for bacteria in biliary lithogenesis. We hypothesized that bacterial DNA is present in gallstones, and that categorical differences exist between gallstone type and the frequency of bacterial sequences. METHODS: Polymerase chain reaction (PCR) was used to amplify bacterial 16S rRNA and uidA (encoding Escherichia coli [E. coli] beta-glucuronidase) genes in different types of gallstones. PCR products were sequenced. RESULTS: Bacterial 16S rRNA and uidA DNA sequences in E. coli were detected in all brown pigment, common bile duct, and mixed cholesterol gallstones (n = 14). In contrast, only one (14%) of seven pure cholesterol gallstones yielded a PCR product. Most (88%) mixed cholesterol gallstones yielded PCR amplification products from their central, as well as their outer, portions. Sequenced products possessed 88-98% identity to 16S rRNA genes of E. coli and Pseudomonas species. CONCLUSIONS: Bacterial DNA sequences are usually present in mixed cholesterol (to 95% cholesterol content), brown pigment, and common bile duct, but rarely in pure cholesterol gallstones. The presence of bacterial beta-glucuronidase is also suggested. The role of bacteria and their products in the formation of mixed cholesterol gallstones, which comprise the majority of cholesterol gallstones, warrants further study.