scholarly journals Interleukin-8 Response in an Intestinal HCT-8 Cell Line Infected with Enteroaggregative and Enterotoxigenic Escherichia coli

2004 ◽  
Vol 11 (3) ◽  
pp. 548-551 ◽  
Author(s):  
David B. Huang ◽  
Herbert L. DuPont ◽  
Zhi-Dong Jiang ◽  
Lily Carlin ◽  
Pablo C. Okhuysen
Keyword(s):  
Escherichia Coli ◽  
Cell Line ◽  
Virulence Factors ◽  
Interleukin 8 ◽  
Enterotoxigenic Escherichia Coli ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infection Model ◽  
E Coli ◽  
Heat Stable ◽  
Line Infection

ABSTRACT This study examined the interleukin-8 (IL-8) response of the intestinal adenocarcinoma HCT-8 cell line to infection with enteroaggregative and enterotoxigenic Escherichia coli pathotypes isolated from patients with travelers' diarrhea. Individual diarrheagenic E. coli strains (enteroaggregative E. coli [EAEC]; n = 30), heat-stable enterotoxin (ST)-producing enterotoxigenic E. coli (ETEC ST; n = 11), heat-labile enterotoxin (LT)-producing enterotoxigenic E. coli (ETEC LT; n = 10), and ST- and LT-producing enterotoxigenic E. coli (ETEC ST:LT; n = 8) were coincubated with HCT-8 cells for 3 h. Tissue culture supernatants were assayed for IL-8 content by enzyme-linked immunosorbent assay. Fifty percent of EAEC (72% of those EAEC carrying the virulence factors aggR, aggA, and aspU and 40% of those EAEC not carrying virulence factors) and 64% of ETEC ST elicited IL-8 production. In contrast, 10% of ETEC LT elicited the production of IL-8 above baseline. These results suggest that (i) the HCT-8 cell line infection model can be used as a tool to differentiate proinflammatory E. coli from noninflammatory isolates; (ii) EAEC has a heterogeneous ability to induce the production of IL-8, and this may be associated with the presence of virulence factors; and (iii) ETEC ST can elicit an inflammatory response and helps explain our earlier findings of increased fecal IL-8 in patients with ETEC diarrhea.

Multiplex Real-Time PCR Detection of Heat-Labile and Heat-Stable Toxin Genes in Enterotoxigenic Escherichia coli †

2006 ◽  
Vol 69 (2) ◽  
pp. 412-416 ◽  
Author(s):  
MICHAEL A. GRANT ◽  
JINXIN HU ◽  
KAREN C. JINNEMAN
Keyword(s):  
Escherichia Coli ◽  
Real Time ◽  
Real Time Pcr ◽  
Membrane Filtration ◽  
Enterotoxigenic Escherichia Coli ◽  
Pcr Analysis ◽  
E Coli ◽  
Toxin Genes ◽  
Heat Stable ◽  
Heat Labile

A multiplex real-time PCR method was developed for detection of heat-labile and heat-stable toxin genes in enterotoxigenic Escherichia coli. Approximately 10 CFU per reaction mixture could be detected in rinsates from produce samples. Several foods representative of varieties previously shown to have caused enterotoxigenic E. coli outbreaks were spiked and enriched for 4 or 6 h. Both heat-labile and heat-stable toxin genes could be detected in the foods tested, with the exception of hot sauce, with threshold cycle values ranging from 25.2 to 41.1. A procedure using membrane filtration which would allow enumeration of the enterotoxigenic E. coli population in a food sample in less than 28 h by real-time PCR analysis of colonies picked from media highly selective for E. coli was also developed.

scholarly journals Molecular Characterization of Shiga Toxin-Producing Escherichia coli Strains Isolated in Poland

2016 ◽  
Vol 65 (3) ◽  
pp. 261-269 ◽  
Author(s):  
Aleksandra Januszkiewicz ◽  
Waldemar Rastawicki
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Shiga Toxin ◽  
Virulence Gene ◽  
Virulence Determinants ◽  
E Coli ◽  
Heat Stable ◽  
Food Borne ◽  
Wide Range ◽  
Uremic Syndrome

Shiga toxin-producing Escherichia coli (STEC) strains also called verotoxin-producing E. coli (VTEC) represent one of the most important groups of food-borne pathogens that can cause several human diseases such as hemorrhagic colitis (HC) and hemolytic – uremic syndrome (HUS) worldwide. The ability of STEC strains to cause disease is associated with the presence of wide range of identified and putative virulence factors including those encoding Shiga toxin. In this study, we examined the distribution of various virulence determinants among STEC strains isolated in Poland from different sources. A total of 71 Shiga toxin-producing E. coli strains isolated from human, cattle and food over the years 1996 – 2010 were characterized by microarray and PCR detection of virulence genes. As stx1a subtype was present in all of the tested Shiga toxin 1 producing E. coli strains, a greater diversity of subtypes was found in the gene stx2, which occurred in five subtypes: stx2a, stx2b, stx2c, stx2d, stx2g. Among STEC O157 strains we observed conserved core set of 14 virulence factors, stable in bacteria genome at long intervals of time. There was one cattle STEC isolate which possessed verotoxin gene as well as sta1 gene encoded heat-stable enterotoxin STIa characteristic for enterotoxigenic E. coli. To the best of our knowledge, this is the first comprehensive analysis of virulence gene profiles identified in STEC strains isolated from human, cattle and food in Poland. The results obtained using microarrays technology confirmed high effectiveness of this method in determining STEC virulotypes which provides data suitable for molecular risk assessment of the potential virulence of this bacteria.

scholarly journals Use of colony pools for diagnosis of enterotoxigenic Escherichia coli diarrhea

1979 ◽  
Vol 9 (4) ◽  
pp. 493-497
Author(s):  
M H Merson ◽  
R B Sack ◽  
A K Kibriya ◽  
A Al-Mahmood ◽  
Q S Adamed ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Enterotoxigenic Escherichia Coli ◽  
Adult Males ◽  
E Coli ◽  
Heat Stable ◽  
Heat Labile ◽  
Stool Specimens ◽  
Stool Cultures ◽  
Made In

Diagnosis of enterotoxigenic Escherichia coli diarrhea was made in 109 adult males with an acute dehydrating cholera-like syndrome in Dacca, Bangladesh, by testing 10 colonies isolated from admission stool specimens for production of heat-labile and heat-stable toxins. Toxin testing of one colony yielded a diagnosis in 92% of the cases, testing of two colonies yielded a diagnosis in 95% of the cases, testing of a pool of 5 colonies yielded a diagnosis in 95% of the cases, and testing of a pool of 10 colonies yielded a diagnosis in 96% of the cases. From stool cultures obtained on subsequent days, toxin testing of individual colonies and pools revealed diminished efficacy of pooling with decreasing numbers of enterotoxin-positive isolates in the pool. To detect the presence of enterotoxigenic E. coli in stools, toxin testing of 5 individual isolates and a pool of 10 colonies was found to be almost as effective as the testing of 10 individual isolates.

scholarly journals Serogroups and virulence genes of Escherichia coli isolated from psittacine birds

2011 ◽  
Vol 31 (10) ◽  
pp. 916-921 ◽  
Author(s):  
Terezinha Knöbl ◽  
André B.S Saidenberg ◽  
Andrea M Moreno ◽  
Tânia A.T Gomes ◽  
Mônica A.M Vieira ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Serum Resistance ◽  
Temperature Sensitive ◽  
E Coli ◽  
Cytotoxic Necrotizing Factor ◽  
Heat Stable ◽  
Genes Encoding ◽  
Cytotoxic Necrotizing Factor 1

Escherichia coli isolates from 24 sick psittacine birds were serogrouped and investigated for the presence of genes encoding the following virulence factors: attaching and effacing (eae), enteropathogenic E. coli EAF plasmid (EAF), pili associated with pyelonephritis (pap), S fimbriae (sfa), afimbrial adhesin (afa), capsule K1 (neu), curli (crl, csgA), temperature-sensitive hemagglutinin (tsh), enteroaggregative heat-stable enterotoxin-1 (astA), heat-stable enterotoxin -1 heat labile (LT) and heat stable (STa and STb) enterotoxins, Shiga-like toxins (stx1 and stx2), cytotoxic necrotizing factor 1 (cnf1), haemolysin (hly), aerobactin production (iuc) and serum resistance (iss). The results showed that the isolates belonged to 12 serogroups: O7; O15; O21; O23; O54; O64; O76; O84; O88; O128; O152 and O166. The virulence genes found were: crl in all isolates, pap in 10 isolates, iss in seven isolates, csgA in five isolates, iuc and tsh in three isolates and eae in two isolates. The combination of virulence genes revealed 11 different genotypic patterns. All strains were negative for genes encoding for EAF, EAEC, K1, sfa, afa, hly, cnf, LT, STa, STb, stx1 and stx2. Our findings showed that some E. coli isolated from psittacine birds present the same virulence factors as avian pathogenic E. coli (APEC), uropathogenic E. coli (UPEC) and Enteropathogenic E. coli (EPEC) pathotypes.

scholarly journals Experimental Infection of Human Volunteers with the Heat-Stable Enterotoxin-Producing Enterotoxigenic Escherichia coli Strain TW11681

Pathogens ◽  
2019 ◽  
Vol 8 (2) ◽  
pp. 84 ◽  
Author(s):  
Sunniva Todnem Sakkestad ◽  
Hans Steinsland ◽  
Steinar Skrede ◽  
Elisabeth Kleppa ◽  
Kristine Lillebø ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
T Cell ◽  
Protective Efficacy ◽  
T Cell Responses ◽  
Enterotoxigenic Escherichia Coli ◽  
Cd4 T Cell ◽  
Infection Model ◽  
Serological Response ◽  
Cell Responses ◽  
Heat Stable

Infection with enterotoxigenic Escherichia coli (ETEC) producing the heat-stable enterotoxin (ST) is one of the most important causes of childhood diarrhoea in low- and middle-income countries. Here, we undertook a controlled human infection model (CHIM) study to investigate whether ST-producing ETEC strain TW11681 would be suitable for testing the protective efficacy of new ST-based vaccine candidates in vaccine challenge models. In groups of three, nine volunteers ingested 1 × 106, 1 × 107, or 1 × 108 colony-forming units (CFU) of TW11681. Flow cytometry-based assays were used to measure CD4+ T cell responses and antibody levels targeting virulence factors expressed by the strain. We found that infection with TW11681 elicited few and mild symptoms, including mild diarrhoea in two volunteers, both of whom ingested 1 × 106 CFU. Averaged across all volunteers, the CD4+ T cell responses specific for E. coli YghJ mucinase peaked 10 days after infection (3.2-fold (p = 0.016)), while the CD4+ T cell responses specific for Colonization Factor Antigen I (CFA/I) major fimbrial subunit (CfaB) peaked after 28 days (3.6-fold (p = 0.063)). The serum CfaB-specific anti-IgA and anti-IgG/IgM levels were significantly increased and peaked 3 months after infection. Both remained elevated for the duration of the 12-month follow-up. The corresponding anti-YghJ serological response was strongest after 10 days, although a significant increase was seen only for IgA levels (3.2-fold (p = 0.008)). In conclusion, due to its low diarrhoea attack risk, TW11681 is probably not suitable for testing the efficacy of new vaccines in human challenge studies at doses 1 × 106 to 1 × 108. However, the strain may still be useful in CHIMs for studying ETEC host-pathogen interactions.

scholarly journals Virulence factors of entericEscherichia coliin young Aboriginal children in north-west Australia

1992 ◽  
Vol 109 (2) ◽  
pp. 283-289 ◽  
Author(s):  
S. T. Gunzburg ◽  
B. J. Chang ◽  
V. Burke ◽  
M. Gracey
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Seasonal Pattern ◽  
Summer Season ◽  
Enteric Pathogens ◽  
Tropical Regions ◽  
North West ◽  
E Coli ◽  
Heat Stable ◽  
Aboriginal Children

SUMMARYEnterotoxigenicEscherichia coli(ETEC) were the most frequently identified enteric pathogens associated with diarrhoea in 0–5 year old Aboriginal children in tropical north-west Australia with an incidence similar to those from other tropical regions. Heat-stable toxin-producing (ST + ) strains were associated with diarrhoea throughout the year but heat-labile toxin-producing (LT + ) strains were more important in the monsoonal summer season. ST + strains were commonest in children with diarrhoea between 6 and 18 months of age while LT + strains were associated with diarrhoea in children aged 18–24 months. VerotoxigenicE. coli(VTEC) which produced VT1, but not VT2, and enteroadherent (EAF + )E. coliwere significant causes of diarrhoea, mainly in children below 18 months but without a seasonal pattern.

scholarly journals The reliability of serogroup determination in the detection of Escherichia coli as a causative agent of sporadic and epidemic occurrence of enterocolitis

2002 ◽  
Vol 59 (3) ◽  
pp. 271-276
Author(s):  
Valentina Stojanovic ◽  
Miloje Cobeljic
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Causative Agent ◽  
Detection Rate ◽  
Reliable Method ◽  
E Coli ◽  
Heat Stable ◽  
Heat Labile ◽  
Sporadic Cases

The purpose of this study was to determine the presence of virulence factors (heat-labile, heat-stable enterotoxin, verotoxin, invasiveness, localized, aggregative and diffuse adherence) among E. coli strains isolated from sporadic cases and outbreaks of enterocolitis, which belonged to serogroups characteristic for enteropathogenic E. coli. Serogroup was determined in 57.2% of 622 strains isolated from sporadic cases, and among them virulence factors were detected in 23.6%. Serogroup was also determined in 73.3% of 90 outbreaks isolates tested and virulence factors were detected in 97% of them. The detection rate of virulence factors rarely exceeded 50% among strains belonging to any of serogroup that was determined. The obtained data suggested that the identification of E. coli as a causative agent of enterocolitis by serogroup determination was a reliable method in outbreaks, but not in sporadic cases of this disease.

scholarly journals Virulence Genes in Expanded-Spectrum-Cephalosporin-Resistant and -Susceptible Escherichia coli Isolates from Treated and Untreated Chickens

2015 ◽  
Vol 60 (3) ◽  
pp. 1874-1877 ◽  
Author(s):  
S. Baron ◽  
S. Delannoy ◽  
S. Bougeard ◽  
E. Larvor ◽  
E. Jouy ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Virulence Factors ◽  
Shiga Toxin ◽  
Virulence Genes ◽  
Phylogenetic Groups ◽  
Content Type ◽  
E Coli ◽  
Heat Stable ◽  
Intestinal Infections

This study investigated antimicrobial resistance, screened for the presence of virulence genes involved in intestinal infections, and determined phylogenetic groups ofEscherichia coliisolates from untreated poultry and poultry treated with ceftiofur, an expanded-spectrum cephalosporin. Results show that none of the 76 isolates appeared to be Shiga toxin-producingE. colior enteropathogenicE. coli. All isolates were negative for the major virulence factors/toxins tested (ehxA,cdt, heat-stable enterotoxin [ST], and heat-labile enterotoxin [LT]). The few virulence genes harbored in isolates generally did not correlate with isolate antimicrobial resistance or treatment status. However, some of the virulence genes were significantly associated with certain phylogenetic groups.

scholarly journals Profiles of virulence genes in enterotoxigenic Escherichia coli isolates from suckling piglets in Serbia

2020 ◽  
Vol 76 (01) ◽  
pp. 6326-2020
Author(s):  
JADRANKA ŽUTIĆ ◽  
OLIVERA VALČIĆ ◽  
VESNA MILIĆEVIĆ ◽  
LJUBIŠA VELJOVIĆ ◽  
JASNA KURELJUŠIĆ ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Virulence Genes ◽  
Enterotoxigenic Escherichia Coli ◽  
Gene Encoding ◽  
E Coli ◽  
Genes Encoding ◽  
Virulence Pattern ◽  
Suckling Piglets ◽  
Asta Gene

A total of 120 Escherichia coli (E. coli) strains from suckling piglets with diarrhoea and 30 E. coli strains from healthy piglets were tested for the presence of fimbrial and enterotoxin virulence genes. Out of the 120 isolates sampled from diarrheic piglets, 81 (67.5%) expressed one or more genes encoding virulence factors. Adhesin genes were detected in 52 (43.33%) out of 120 E. coli isolates, and the most common among them was F4 adhesin (33.33%). Genes encoding E. coli toxins were detected in 81 (67.5%) isolates. E. coli included in the study carried genes for one or more of the following toxins: STa, STb, LT and EAST1. The astA gene encoding EAST1 was the most prevalent and was identified in 72 (60%) E. coli isolates. EAST1 toxin was detected in 5 out of 30 isolates (16.7%) from healthy piglets. Among the 81 isolates expressing virulence genes, a total of 15 different combinations for fimbrial and toxin genes were found. The most common virulence pattern was F4/STb/LT/EAST1 detected in 23.45% of E. coli strains isolated from suckling piglets with diarrhoea. The results indicate that F4 adhesin and EAST1 toxin are the most common in E. coli isolates sampled from diarrhoeic suckling piglets in Serbia.

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