Characterization of enterohemorrhagicEscherichia coliO157:H7 00B015: a Shiga toxin producing but virulence-attenuated isolate

2010 ◽  
Vol 56 (8) ◽  
pp. 651-656 ◽  
Author(s):  
Haiguang Wang ◽  
Jiang Gu ◽  
Shu Yu ◽  
Weijun Zhang ◽  
Yefei Zhu ◽  
...  

Enterohemorrhagic Escherichia coli (EHEC) causes a wide range of systematic diseases in human and animals in 2 main ways: (1) production of Shiga toxin (Stx) and (2) induction of actin polymerization characterized by attaching and effacing (A/E) lesions. Stx is commonly targeted in the development of drugs and vaccines to control EHEC infection for its indispensible contribution to EHEC pathogenisis. In this study, we isolated a Stx-producing EHEC O157:H7 isolate 00B015 and found that its ability to induce actin polymerization was impaired. In addition, it reduces pathogenicity and decreases mortality in mice. Our results report a Stx-producing but virulence-attenuated EHEC isolate 00B015 and suggest that the formation of actin polymerization may help Stx-induced pathogenesis and have a more important contribution in EHEC infections.

2013 ◽  
Vol 7 (21) ◽  
pp. 2637-2645 ◽  
Author(s):  
M EL Alfy Sahar ◽  
F Ahmed Salwa ◽  
A Selim Samy ◽  
H Abdel Aziz Mohamed ◽  
M Zakaria Amira ◽  
...  

Toxins ◽  
2021 ◽  
Vol 13 (2) ◽  
pp. 139
Author(s):  
Johanna Detzner ◽  
Elisabeth Krojnewski ◽  
Gottfried Pohlentz ◽  
Daniel Steil ◽  
Hans-Ulrich Humpf ◽  
...  

Human kidney epithelial cells are supposed to be directly involved in the pathogenesis of the hemolytic–uremic syndrome (HUS) caused by Shiga toxin (Stx)-producing enterohemorrhagic Escherichia coli (EHEC). The characterization of the major and minor Stx-binding glycosphingolipids (GSLs) globotriaosylceramide (Gb3Cer) and globotetraosylceramide (Gb4Cer), respectively, of primary human renal cortical epithelial cells (pHRCEpiCs) revealed GSLs with Cer (d18:1, C16:0), Cer (d18:1, C22:0), and Cer (d18:1, C24:1/C24:0) as the dominant lipoforms. Using detergent-resistant membranes (DRMs) and non-DRMs, Gb3Cer and Gb4Cer prevailed in the DRM fractions, suggesting their association with microdomains in the liquid-ordered membrane phase. A preference of Gb3Cer and Gb4Cer endowed with C24:0 fatty acid accompanied by minor monounsaturated C24:1-harboring counterparts was observed in DRMs, whereas the C24:1 fatty acid increased in relation to the saturated equivalents in non-DRMs. A shift of the dominant phospholipid phosphatidylcholine with saturated fatty acids in the DRM to unsaturated species in the non-DRM fractions correlated with the GSL distribution. Cytotoxicity assays gave a moderate susceptibility of pHRCEpiCs to the Stx1a and Stx2a subtypes when compared to highly sensitive Vero-B4 cells. The results indicate that presence of Stx-binding GSLs per se and preferred occurrence in microdomains do not necessarily lead to a high cellular susceptibility towards Stx.


Virulence ◽  
2021 ◽  
Vol 12 (1) ◽  
pp. 1296-1305
Author(s):  
Ying Hua ◽  
Milan Chromek ◽  
Anne Frykman ◽  
Cecilia Jernberg ◽  
Valya Georgieva ◽  
...  

2014 ◽  
Vol 77 (7) ◽  
pp. 1212-1218 ◽  
Author(s):  
BURTON BLAIS ◽  
MYLÈNE DESCHÊNES ◽  
GEORGE HUSZCZYNSKI ◽  
MARTINE GAUTHIER

A simple immunoenzymatic enterohemorrhagic Escherichia coli (EHEC) colony check (ECC) assay was developed for the presumptive identification of priority EHEC colonies isolated on plating media from enrichment broth cultures of foods. With this approach, lipopolysaccharide extracted from a colony is spotted on the grid of a polymyxin-coated polyester cloth strip, and bound E. coli serogroup O26, O45, O103, O111, O121, O145, and O157 antigens are subsequently detected by sequential reactions with a pool of commercially available peroxidase-conjugated goat antibodies and tetramethylbenzidine substrate solution. Each strip can accommodate up to 15 colonies, and test results are available within 30 min. Assay performance was verified using colonies from a total of 73 target EHEC isolates covering the range of designated priority serogroups (all of which were reactive), 41 nontarget E. coli isolates including several nontarget Shiga toxin–producing E. coli serogroups (all unreactive), and 33 non–E. coli strains (all unreactive except two bacterial strains possessing O-antigenic structures in common with those of the priority EHEC). The ECC assay was reactive with target colonies grown on several types of selective and nonselective plating media designed for their cultivation. These results support the use of the ECC assay for high-throughput screening of colonies isolated on plating media for detecting priority EHEC strains in foods.


Viruses ◽  
2018 ◽  
Vol 10 (11) ◽  
pp. 595 ◽  
Author(s):  
Ling Zhang ◽  
David Simpson ◽  
Lynn McMullen ◽  
Michael Gänzle

Shiga-toxin producing Escherichia coli (STEC) causes human illness ranging from mild diarrhea to death. The bacteriophage encoded stx genes are located in the late transcription region, downstream of the antiterminator Q. The transcription of the stx genes is directly under the control of the late promoter pR’, thus the sequence diversity of the region between Q and stx, here termed the pR’ region, may affect Stx toxin production. Here, we compared the gene structure of the pR’ region and the stx subtypes of nineteen STECs. The sequence alignment and phylogenetic analysis suggested that the pR’ region tends to be more heterogeneous than the promoter itself, even if the prophages harbor the same stx subtype. Furthermore, we established and validated transcriptional fusions of the pR’ region to the DsRed reporter gene using mitomycin C (MMC) induction. Finally, these constructs were transformed into native and non-native strains and examined with flow cytometry. The results showed that induction levels changed when pR’ regions were placed under different regulatory systems. Moreover, not every stx gene could be induced in its native host bacteria. In addition to the functional genes, the diversity of the pR’ region plays an important role in determining the level of toxin induction.


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