Comparison of Asymptomatic Bacteriuria Escherichia coli Isolates from Healthy Individuals versus Those from Hospital Patients Shows that Long-Term Bladder Colonization Selects for Attenuated Virulence Phenotypes

ABSTRACTAsymptomatic bacteriuria (ABU) is a condition where bacteria stably colonize the urinary tract, in a manner closely resembling commensalism at other mucosal sites. The patients carry >105CFU/ml for extended periods of time and rarely develop symptoms. Contrasting the properties of ABU strains to those of uropathogenic isolates causing symptomatic infection is therefore highly relevant to understand mechanisms of bacterial adaptation. The prototype ABU strainEscherichia coli83972 has a smaller genome than uropathogenicE. coli(UPEC) strains with deletions or point mutations in several virulence genes, suggesting that ABU strains undergo a programmed reductive evolution within human hosts. This study addressed if these observations can be generalized. Strains causing ABU in outpatients or hospitalized patients after catheterization or other invasive procedures were compared to commensalE. coliisolates from the intestinal flora of healthy individuals. Notably, clonal complex 73 (CC73) was a prominent phylogenetic lineage dominated by ABU isolates. ABU isolates from outpatients and hospitalized patients had a similar overall virulence gene repertoire, which distinguished them from many commensals, but typical UPEC virulence genes were less frequently attenuated in hospital strains than in outpatient strains or commensals. The decreased virulence potential of outpatient ABU isolates relative to that of ABU strains from hospitalized patients supports the hypothesis that loss of expression or decay of virulence genes facilitates long-term carriage and adaptation to host environments.

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Escherichia coli EDL933 Requires Gluconeogenic Nutrients To Successfully Colonize the Intestines of Streptomycin-Treated Mice Precolonized with E. coli Nissle 1917

Infection and Immunity ◽

10.1128/iai.02943-14 ◽

2015 ◽

Vol 83 (5) ◽

pp. 1983-1991 ◽

Cited By ~ 12

Author(s):

Silvia A. C. Schinner ◽

Matthew E. Mokszycki ◽

Jimmy Adediran ◽

Mary Leatham-Jensen ◽

Tyrrell Conway ◽

...

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Probiotic Strain ◽

Content Type ◽

E Coli ◽

Link Type ◽

Mouse Intestine ◽

K 12 ◽

Rule Out

Escherichia coliMG1655, a K-12 strain, uses glycolytic nutrients exclusively to colonize the intestines of streptomycin-treated mice when it is the onlyE. colistrain present or when it is confronted withE. coliEDL933, an O157:H7 strain. In contrast,E. coliEDL933 uses glycolytic nutrients exclusively when it is the onlyE. colistrain in the intestine but switches in part to gluconeogenic nutrients when it colonizes mice precolonized withE. coliMG1655 (R. L. Miranda et al., Infect Immun 72:1666–1676, 2004,http://dx.doi.org/10.1128/IAI.72.3.1666-1676.2004). Recently, J. W. Njoroge et al. (mBio 3:e00280-12, 2012,http://dx.doi.org/10.1128/mBio.00280-12) reported thatE. coli86-24, an O157:H7 strain, activates the expression of virulence genes under gluconeogenic conditions, suggesting that colonization of the intestine with a probioticE. colistrain that outcompetes O157:H7 strains for gluconeogenic nutrients could render them nonpathogenic. Here we report thatE. coliNissle 1917, a probiotic strain, uses both glycolytic and gluconeogenic nutrients to colonize the mouse intestine between 1 and 5 days postfeeding, appears to stop using gluconeogenic nutrients thereafter in a large, long-term colonization niche, but continues to use them in a smaller niche to compete with invadingE. coliEDL933. Evidence is also presented suggesting that invadingE. coliEDL933 uses both glycolytic and gluconeogenic nutrients and needs the ability to perform gluconeogenesis in order to colonize mice precolonized withE. coliNissle 1917. The data presented here therefore rule out the possibility thatE. coliNissle 1917 can starve the O157:H7E. colistrain EDL933 of gluconeogenic nutrients, even thoughE. coliNissle 1917 uses such nutrients to compete withE. coliEDL933 in the mouse intestine.

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Subtilase Cytotoxin-Coding Genes in Verotoxin-Producing Escherichia coli Strains from Sheep and Goats Differ from Those from Cattle

Applied and Environmental Microbiology ◽

10.1128/aem.05604-11 ◽

2011 ◽

Vol 77 (23) ◽

pp. 8259-8264 ◽

Cited By ~ 11

Author(s):

José A. Orden ◽

Pilar Horcajo ◽

Ricardo de la Fuente ◽

José A. Ruiz-Santa-Quiteria ◽

Gustavo Domínguez-Bernal ◽

...

Keyword(s):

Escherichia Coli ◽

Human Disease ◽

Virulence Genes ◽

Small Ruminants ◽

Human Beings ◽

Content Type ◽

Sheep And Goats ◽

E Coli ◽

Healthy Cattle ◽

Subtilase Cytotoxin

ABSTRACTSubtilase cytotoxin (SubAB) from verotoxin (VT)-producingEscherichia coli(VTEC) strains was first described in the 98NK2 strain and has been associated with human disease. However, SubAB has recently been found in two VT-negativeE. colistrains (ED 591 and ED 32). SubAB is encoded by two closely linked, cotranscribed genes (subAandsubB). In this study, we investigated the presence ofsubABgenes in 52 VTEC strains isolated from cattle and 209 strains from small ruminants, using PCR. Most (91.9%) VTEC strains from sheep and goats and 25% of the strains from healthy cattle possessedsubABgenes. The presence ofsubABin a high percentage of the VTEC strains from small ruminants might increase the pathogenicity of these strains for human beings. Some differences in the results of PCRs and in the association with some virulence genes suggested the existence of different variants ofsubAB. We therefore sequenced thesubAgene in 12 strains and showed that thesubAgene in most of thesubAB-positive VTEC strains from cattle was almost identical (about 99%) to that in the 98NK2 strain, while thesubAgene in most of thesubAB-positive VTEC strains from small ruminants was almost identical to that in the ED 591 strain. We propose the termssubAB1to describe the SubAB-coding genes resembling that in the 98NK2 strain andsubAB2to describe those resembling that in the ED 591 strain.

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Proteorhodopsin Overproduction Enhances the Long-Term Viability of Escherichia coli

Applied and Environmental Microbiology ◽

10.1128/aem.02087-19 ◽

2019 ◽

Vol 86 (1) ◽

Cited By ~ 2

Author(s):

Yizhi Song ◽

Michaël L. Cartron ◽

Philip J. Jackson ◽

Paul A. Davison ◽

Mark J. Dickman ◽

...

Keyword(s):

Escherichia Coli ◽

Raman Spectroscopy ◽

Cell Membrane ◽

Single Cell ◽

Marine Bacteria ◽

Membrane Area ◽

Content Type ◽

E Coli ◽

Wide Range

ABSTRACT Genes encoding the photoreactive protein proteorhodopsin (PR) have been found in a wide range of marine bacterial species, reflecting the significant contribution that PR makes to energy flux and carbon cycling in ocean ecosystems. PR can also confer advantages to enhance the ability of marine bacteria to survive periods of starvation. Here, we investigate the effect of heterologously produced PR on the viability of Escherichia coli. Quantitative mass spectrometry shows that E. coli, exogenously supplied with the retinal cofactor, assembles as many as 187,000 holo-PR molecules per cell, accounting for approximately 47% of the membrane area; even cells with no retinal synthesize ∼148,000 apo-PR molecules per cell. We show that populations of E. coli cells containing PR exhibit significantly extended viability over many weeks, and we use single-cell Raman spectroscopy (SCRS) to detect holo-PR in 9-month-old cells. SCRS shows that such cells, even incubated in the dark and therefore with inactive PR, maintain cellular levels of DNA and RNA and avoid deterioration of the cytoplasmic membrane, a likely basis for extended viability. The substantial proportion of the E. coli membrane required to accommodate high levels of PR likely fosters extensive intermolecular contacts, suggested to physically stabilize the cell membrane and impart a long-term benefit manifested as extended viability in the dark. We propose that marine bacteria could benefit similarly from a high PR content, with a stabilized cell membrane extending survival when those bacteria experience periods of severe nutrient or light limitation in the oceans. IMPORTANCE Proteorhodopsin (PR) is part of a diverse, abundant, and widespread superfamily of photoreactive proteins, the microbial rhodopsins. PR, a light-driven proton pump, enhances the ability of the marine bacterium Vibrio strain AND4 to survive and recover from periods of starvation, and heterologously produced PR extends the viability of nutrient-limited Shewanella oneidensis. We show that heterologously produced PR enhances the viability of E. coli cultures over long periods of several weeks and use single-cell Raman spectroscopy (SCRS) to detect PR in 9-month-old cells. We identify a densely packed and consequently stabilized cell membrane as the likely basis for extended viability. Similar considerations are suggested to apply to marine bacteria, for which high PR levels represent a significant investment in scarce metabolic resources. PR-stabilized cell membranes in marine bacteria are proposed to keep a population viable during extended periods of light or nutrient limitation, until conditions improve.

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Spread of NDM-5 and OXA-181 Carbapenemase-Producing Escherichia coli in Chad

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.00646-19 ◽

2019 ◽

Vol 63 (11) ◽

Cited By ~ 2

Author(s):

Oumar Ouchar Mahamat ◽

Manon Lounnas ◽

Mallorie Hide ◽

Abelsalam Tidjani ◽

Julio Benavides ◽

...

Keyword(s):

Escherichia Coli ◽

Healthy Volunteers ◽

Resistance Genes ◽

Sequence Type ◽

Hospitalized Patients ◽

Content Type ◽

E Coli ◽

First Time

ABSTRACT We detected for the first time blaNDM-5 and blaOXA-181 in Escherichia coli isolates from hospitalized patients and healthy volunteers in Chad. These resistance genes were located on IncX3 and IncF plasmids. Despite the large diversity of E. coli clones, the identified resistant intestinal isolates belonged mainly to the same sequence type.

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Extraintestinal Pathogenic and Antimicrobial-Resistant Escherichia coli, Including Sequence Type 131 (ST131), from Retail Chicken Breasts in the United States in 2013

Applied and Environmental Microbiology ◽

10.1128/aem.02956-16 ◽

2017 ◽

Vol 83 (6) ◽

Cited By ~ 15

Author(s):

James R. Johnson ◽

Stephen B. Porter ◽

Brian Johnston ◽

Paul Thuras ◽

Sarah Clock ◽

...

Keyword(s):

United States ◽

Escherichia Coli ◽

Virulence Genes ◽

Meat Products ◽

The United States ◽

Sequence Type ◽

The Other ◽

Antibiotic Resistant ◽

Content Type ◽

E Coli

ABSTRACT Chicken meat products are hypothesized to be vehicles for transmitting antimicrobial-resistant and extraintestinal pathogenic Escherichia coli (ExPEC) to consumers. To reassess this hypothesis in the current era of heightened concerns about antimicrobial use in food animals, we analyzed 175 chicken-source E. coli isolates from a 2013 Consumer Reports national survey. Isolates were screened by PCR for ExPEC-defining virulence genes. The 25 ExPEC isolates (12% of 175) and a 2:1 randomly selected set of 50 non-ExPEC isolates were assessed for their phylogenetic/clonal backgrounds and virulence genotypes for comparison with their resistance profiles and the claims on the retail packaging label (“organic,” “no antibiotics,” and “natural”). Compared with the findings for non-ExPEC isolates, the group of ExPEC isolates had a higher prevalence of phylogroup B2 isolates (44% versus 4%; P < 0.001) and a lower prevalence of phylogroup A isolates (4% versus 30%; P = 0.001), a higher prevalence of multiple individual virulence genes, higher virulence scores (median, 11 [range, 4 to 16] versus 8 [range, 1 to 14]; P = 0.001), and higher resistance scores (median, 4 [range, 0 to 8] versus 3 [range, 0 to 10]; P < 0.001). All five isolates of sequence type 131 (ST131) were ExPEC (P = 0.003), were as extensively resistant as the other isolates tested, and had higher virulence scores than the other isolates tested (median, 12 [range, 11 to 13] versus 8 [range, 1 to 16]; P = 0.005). Organic labeling predicted lower resistance scores (median, 2 [range, 0 to 3] versus 4 [range, 0 to 10]; P = 0.008) but no difference in ExPEC status or virulence scores. These findings document a persisting reservoir of extensively antimicrobial-resistant ExPEC isolates, including isolates from ST131, in retail chicken products in the United States, suggesting a potential public health threat. IMPORTANCE We found that among Escherichia coli isolates from retail chicken meat products purchased across the United States in 2013 (many of these isolates being extensively antibiotic resistant), a minority had genetic profiles suggesting an ability to cause extraintestinal infections in humans, such as urinary tract infection, implying a risk of foodborne disease. Although isolates from products labeled “organic” were less extensively antibiotic resistant than other isolates, they did not appear to be less virulent. These findings suggest that retail chicken products in the United States, even if they are labeled “organic,” pose a potential health threat to consumers because they are contaminated with extensively antibiotic-resistant and, presumably, virulent E. coli isolates.

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Estimating the Prevalence of Potential Enteropathogenic Escherichia coli and Intimin Gene Diversity in a Human Community by Monitoring Sanitary Sewage

Applied and Environmental Microbiology ◽

10.1128/aem.02747-13 ◽

2013 ◽

Vol 80 (1) ◽

pp. 119-127 ◽

Cited By ~ 12

Author(s):

Kun Yang ◽

Eulyn Pagaling ◽

Tao Yan

Keyword(s):

Escherichia Coli ◽

Virulence Genes ◽

Gene Diversity ◽

Content Type ◽

E Coli ◽

Enteric Diseases ◽

Detection Frequency ◽

Alternative Approach ◽

Prevalent Subtype ◽

Qpcr Quantification

ABSTRACTPresently, the understanding of bacterial enteric diseases in the community and their virulence factors relies almost exclusively on clinical disease reporting and examination of clinical pathogen isolates. This study aimed to investigate the feasibility of an alternative approach that monitors potential enteropathogenicEscherichia coli(EPEC) and enterohemorrhagicE. coli(EHEC) prevalence and intimin gene (eae) diversity in a community by directly quantifying and characterizing target virulence genes in the sanitary sewage. The quantitative PCR (qPCR) quantification of theeae,stx1, andstx2genes in sanitary sewage samples collected over a 13-month period detectedeaein all 13 monthly sewage samples at significantly higher abundance (93 to 7,240 calibrator cell equivalents [CCE]/100 ml) thanstx1andstx2, which were detected sporadically. The prevalence level of potential EPEC in the sanitary sewage was estimated by calculating the ratio ofeaetouidA, which averaged 1.0% (σ = 0.4%) over the 13-month period. Cloning and sequencing of theeaegene directly from the sewage samples covered the majority of theeaediversity in the sewage and detected 17 uniqueeaealleles belonging to 14 subtypes. Among them,eae-β2 was identified to be the most prevalent subtype in the sewage, with the highest detection frequency in the clone libraries (41.2%) and within the different sampling months (85.7%). Additionally, sewage and environmentalE. coliisolates were also obtained and used to determine the detection frequencies of the virulence genes as well aseaegenetic diversity for comparison.

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Molecular Epidemiology and Genome Dynamics of New Delhi Metallo-β-Lactamase-Producing Extraintestinal Pathogenic Escherichia coli Strains from India

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.01345-16 ◽

2016 ◽

Vol 60 (11) ◽

pp. 6795-6805 ◽

Cited By ~ 32

Author(s):

Amit Ranjan ◽

Sabiha Shaik ◽

Agnismita Mondal ◽

Nishant Nandanwar ◽

Arif Hussain ◽

...

Keyword(s):

Escherichia Coli ◽

Effective Means ◽

Genomic Analysis ◽

Virulence Gene ◽

Carbapenem Resistance ◽

Control Measures ◽

Large Sample Size ◽

Gene Repertoire ◽

Content Type ◽

E Coli

ABSTRACTThe global dissemination and increasing incidence of carbapenem-resistant, Gram-negative organisms have resulted in acute public health concerns. Here, we present a retrospective multicenter study on molecular characterization of metallo-β-lactamase (MBL)-producing clinicalEscherichia coliisolates recovered from extraintestinal infections in two hospitals in Pune, India. We screened a large sample size of 510E. coliisolates for MBL production wherein we profiled their molecular determinants, antimicrobial resistance phenotypes, functional virulence properties, genomic features, and transmission dynamics. Approximately 8% of these isolates were MBL producers, the majority of which were of the NDM-1 (69%) type, followed by NDM-5 (19%), NDM-4 (5.5%), and NDM-7 (5.5%). MBL producers were resistant to all antibiotics tested except for colistin, fosfomycin, and chloramphenicol, which were effective to various extents. Plasmids were found to be an effective means of dissemination of NDM genes and other resistance traits. All MBL producers adhered to and invaded bladder epithelial (T24) cells and demonstrated significant serum resistance. Genomic analysis of MBL-producingE. coliisolates revealed higher resistance but a moderate virulence gene repertoire. A subset of NDM-1-positiveE. coliisolates was identified as dominant sequence type 101 (ST101) while two strains belonging to ST167 and ST405 harbored NDM-5. A majority of MBL-producingE. colistrains revealed unique genotypes, suggesting that they were clonally unrelated. Overall, the coexistence of virulence and carbapenem resistance in clinicalE. coliisolates is of serious concern. Moreover, the emergence of NDM-1 among the globally dominantE. coliST101 isolates warrants stringent surveillance and control measures.

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Extended-Spectrum-β-Lactamase- and Plasmid AmpC-Producing Escherichia coli Causing Community-Onset Bloodstream Infection: Association of Bacterial Clones and Virulence Genes with Septic Shock, Source of Infection, and Recurrence

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.02351-19 ◽

2020 ◽

Vol 64 (8) ◽

Author(s):

Inga Fröding ◽

Badrul Hasan ◽

Isak Sylvin ◽

Maarten Coorens ◽

Pontus Nauclér ◽

...

Keyword(s):

Escherichia Coli ◽

Septic Shock ◽

Prostate Biopsy ◽

Virulence Genes ◽

Multivariable Analysis ◽

Bloodstream Infections ◽

Content Type ◽

E Coli ◽

Extended Spectrum ◽

Community Onset

ABSTRACT Invasive infections due to extended-spectrum-β-lactamase- and pAmpC-producing Escherichia coli (ESBL/pAmpC-EC) are an important cause of morbidity, often caused by the high-risk clone sequence type (ST131) and isolates classified as extraintestinal pathogenic E. coli (ExPEC). The relative influence of host immunocompetence versus microbiological virulence factors in the acquisition and outcome of bloodstream infections (BSI) is poorly understood. Herein, we used whole-genome sequencing on 278 blood culture isolates of ESBL/pAmpC-EC from 260 patients with community-onset BSI collected from 2012 to 2015 in Stockholm to study the association of virulence genes, sequence types, and antimicrobial resistance with severity of disease, infection source, ESBL/pAmpC-EC BSI low-risk patients, and patients with repeated episodes. ST131 subclade C2 comprised 29% of all patients. Factors associated with septic shock in multivariable analysis were patient host factors (hematologic cancer or transplantation and reduced daily living activity), presence of the E. coli virulence factor iss (increased serum survival), absence of phenotypic multidrug resistance, and absence of the genes pap and hsp. Adhesins, particularly pap, were associated with urinary tract infection (UTI) source, while isolates from post-prostate biopsy sepsis had a low overall number of virulence operons, including adhesins, and commonly belonged to ST131 clades A, B, and subclade C1, ST1193, and ST648. ST131 was associated with recurrent episodes. In conclusion, the most interesting finding is the association of iss with septic shock. Adhesins are important for UTI pathogenesis, while otherwise low-pathogenic isolates from the microbiota can cause post-prostate biopsy sepsis.

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Transcriptional Responses of Escherichia coli K-12 and O157:H7 Associated with Lettuce Leaves

Applied and Environmental Microbiology ◽

10.1128/aem.07454-11 ◽

2012 ◽

Vol 78 (6) ◽

pp. 1752-1764 ◽

Cited By ~ 64

Author(s):

Ryan C. Fink ◽

Elaine P. Black ◽

Zhe Hou ◽

Masayuki Sugawara ◽

Michael J. Sadowsky ◽

...

Keyword(s):

Escherichia Coli ◽

Molecular Mechanisms ◽

Leaf Surface ◽

Short Term ◽

Transcriptional Responses ◽

Content Type ◽

E Coli ◽

Leaf Surfaces ◽

K 12

ABSTRACTAn increasing number of outbreaks of gastroenteritis recently caused byEscherichia coliO157:H7 have been linked to the consumption of leafy green vegetables. Although it is known thatE. colisurvives and grows in the phyllosphere of lettuce plants, the molecular mechanisms by which this bacterium associates with plants are largely unknown. The goal of this study was to identifyE. coligenes relevant to its interaction, survival, or attachment to lettuce leaf surfaces, comparingE. coliK-12, a model system, andE. coliO157:H7, a pathogen associated with a large number of outbreaks. Using microarrays, we found that upon interaction with intact leaves, 10.1% and 8.7% of the 3,798 shared genes were differentially expressed in K-12 and O157:H7, respectively, whereas 3.1% changed transcript levels in both. The largest group of genes downregulated consisted of those involved in energy metabolism, includingtnaA(33-fold change), encoding a tryptophanase that converts tryptophan into indole. Genes involved in biofilm modulation (bhsAandybiM) and curli production (csgAandcsgB) were significantly upregulated inE. coliK-12 and O157:H7. BothcsgAandbhsA(ycfR) mutants were impaired in the long-term colonization of the leaf surface, but onlycsgAmutants had diminished ability in short-term attachment experiments. Our data suggested that the interaction ofE. coliK-12 and O157:H7 with undamaged lettuce leaves likely is initiated via attachment to the leaf surface using curli fibers, a downward shift in their metabolism, and the suppression of biofilm formation.

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Occurrence of Virulence Genes Associated with Diarrheagenic Pathotypes in Escherichia coli Isolates from Surface Water

Applied and Environmental Microbiology ◽

10.1128/aem.02888-12 ◽

2012 ◽

Vol 79 (1) ◽

pp. 328-335 ◽

Cited By ~ 41

Author(s):

Jatinder P. S. Sidhu ◽

Warish Ahmed ◽

Leonie Hodgers ◽

Simon Toze

Keyword(s):

Escherichia Coli ◽

Health Risks ◽

High Frequency ◽

Aquatic Ecosystems ◽

Virulence Genes ◽

Storm Water ◽

Storm Events ◽

Widespread Occurrence ◽

Content Type ◽

E Coli

ABSTRACTEscherichia coliisolates (n= 300) collected from six sites in subtropical Brisbane, Australia, prior to and after storm events were tested for the presence of 11 virulence genes (VGs) specific to diarrheagenic pathotypes. The presence ofeaeA,stx1,stx2, andehxAgenes specific for the enterohemorrhagicE. coli(EHEC) pathotype was detected in 56%, 6%, 10%, and 13% of isolates, respectively. The VGsastA(69%) andaggR(29%), carried by enteroaggregative (EAEC) pathotypes, were frequently detected inE. coliisolates. The enteropathogenicE. coli(EPEC) genebfpwas detected in 24% of isolates. In addition, enteroinvasiveE. coli(EIEC) VGipaHwas also detected in 14% of isolates. During dry periods, isolates belonging to the EAEC pathotype were most commonly detected (23%), followed by EHEC (11%) and EPEC (11%). Conversely, a more uniform prevalence of pathotypes, EPEC (14%), EAEC (12%), EIEC (10%), EHEC (7%), and ETEC (7%), was observed after the storm events. The results of this study highlight the widespread occurrence of potentially diarrheagenic pathotypes in the urban aquatic ecosystems. While the presence of VGs inE. coliisolates alone is insufficient to determine pathogenicity, the presence of diarrheagenicE. colipathotypes in high frequency after the storm events could lead to increased health risks if untreated storm water were to be used for nonpotable purposes and recreational activities.

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