scholarly journals The impact of acid stress on escherichia coli O157:H7 virulence

2021 ◽  
Author(s):  
Belinda House
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cells ◽  
Host Cell ◽  
Virulence Factors ◽  
Acid Stress ◽  
Mrna Levels ◽  
E Coli ◽  
Acid Shock ◽  
The Impact

Escherichia coli 0157:H7 infection is a leading cause of hemorrhagic colitis, and hemolytic uremic syndrome. Many opportunities for acid stress exposure exist for this food and waterborne pathogen, including gastric acid shock. Yet little is known how this affects E.coli 0157:H7 virulence. The effect of various acid stress protocols on E. coli 0157:H7 survival, verotoxin production, and the ability to adhere to host epithelial cells was examined. Brief acid shock alone at pH 3.0 decreased the host cell adhesion capability by a factor of 4.3-4.8, yet when the acid shock was preceded by adaptation at pH 5.0, a 1.6-3.2 fold enhanced adhesion of surviving organisms to epithelial cells relative to unstressed organisms was observed. However, acid stress did not affect verotoxin production. Pretreatment of acid stressed bacteria with erythromycin eliminated the acid-induced adhesion enhancement, suggesting that protein synthesis is a requirement for the enhanced adhesion observed with acid-adapted acid-shocked E.coli 0157:H7. Real time PCR analysis of locus for enterocyte effacement (LEE)-encoded virulence factors, intimin and EspA, revealed no significant upregulation for the acid stress treatments associated with the increased host cell adhesion. On the contrary, elevated mRNA levels for both intimin and EspA were observed for bacteria subjected to brief acid shock alone even though the host-cell adhesion was significatly decreased with these treatments. These results suggest that complex regulation mechanisms for LEE encoded virulence factors exists and that E. coli 0157:H7 virulence can be enhanced after acid stress through increased adhesion to host epithelial cells.

scholarly journals The impact of acid stress on escherichia coli O157:H7 virulence

2021 ◽  
Author(s):  
Belinda House
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cells ◽  
Host Cell ◽  
Virulence Factors ◽  
Acid Stress ◽  
Mrna Levels ◽  
E Coli ◽  
Acid Shock ◽  
The Impact

Escherichia coli 0157:H7 infection is a leading cause of hemorrhagic colitis, and hemolytic uremic syndrome. Many opportunities for acid stress exposure exist for this food and waterborne pathogen, including gastric acid shock. Yet little is known how this affects E.coli 0157:H7 virulence. The effect of various acid stress protocols on E. coli 0157:H7 survival, verotoxin production, and the ability to adhere to host epithelial cells was examined. Brief acid shock alone at pH 3.0 decreased the host cell adhesion capability by a factor of 4.3-4.8, yet when the acid shock was preceded by adaptation at pH 5.0, a 1.6-3.2 fold enhanced adhesion of surviving organisms to epithelial cells relative to unstressed organisms was observed. However, acid stress did not affect verotoxin production. Pretreatment of acid stressed bacteria with erythromycin eliminated the acid-induced adhesion enhancement, suggesting that protein synthesis is a requirement for the enhanced adhesion observed with acid-adapted acid-shocked E.coli 0157:H7. Real time PCR analysis of locus for enterocyte effacement (LEE)-encoded virulence factors, intimin and EspA, revealed no significant upregulation for the acid stress treatments associated with the increased host cell adhesion. On the contrary, elevated mRNA levels for both intimin and EspA were observed for bacteria subjected to brief acid shock alone even though the host-cell adhesion was significatly decreased with these treatments. These results suggest that complex regulation mechanisms for LEE encoded virulence factors exists and that E. coli 0157:H7 virulence can be enhanced after acid stress through increased adhesion to host epithelial cells.

scholarly journals Acid-stress-induced changes in enterohaemorrhagic Escherichia coli O157 : H7 virulence

Microbiology ◽  
2009 ◽  
Vol 155 (9) ◽  
pp. 2907-2918 ◽  
Author(s):  
B. House ◽  
J. V. Kus ◽  
N. Prayitno ◽  
R. Mair ◽  
L. Que ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Host Cell ◽  
Shiga Toxin ◽  
Cell Apoptosis ◽  
Acid Stress ◽  
Enterohaemorrhagic Escherichia Coli ◽  
Host Cell Apoptosis ◽  
Acid Shock ◽  
The Impact ◽  
Virulence Properties

Enterohaemorrhagic Escherichia coli (EHEC) O157 : H7 is naturally exposed to a wide variety of stresses including gastric acid shock, and yet little is known about how this stress influences virulence. This study investigated the impact of acid stress on several critical virulence properties including survival, host adhesion, Shiga toxin production, motility and induction of host-cell apoptosis. Several acid-stress protocols with relevance for gastric passage as well as external environmental exposure were included. Acute acid stress at pH 3 preceded by acid adaptation at pH 5 significantly enhanced the adhesion of surviving organisms to epithelial cells and bacterial induction of host-cell apoptosis. Motility was also significantly increased after acute acid stress. Interestingly, neither secreted nor periplasmic levels of Shiga toxin were affected by acid shock. Pretreatment of bacteria with erythromycin eliminated the acid-induced adhesion enhancement, suggesting that de novo protein synthesis was required for the enhanced adhesion of acid-shocked organisms. DNA microarray was used to analyse the transcriptome of an EHEC O157 : H7 strain exposed to three different acid-stress treatments. Expression profiles of acid-stressed EHEC revealed significant changes in virulence factors associated with adhesion, motility and type III secretion. These results document profound changes in the virulence properties of EHEC O157 : H7 after acid stress, provide a comprehensive genetic analysis to substantiate these changes and suggest strategies that this pathogen may use during gastric passage and colonization in the human gastrointestinal tract.

scholarly journals Solubility assessment of single-chain antibody fragment against epithelial cell adhesion molecule extracellular domain in four Escherichia coli strains

2021 ◽  
Vol 19 (1) ◽  
Author(s):  
Fatemeh Sadat Javadian ◽  
Majid Basafa ◽  
Aidin Behravan ◽  
Atieh Hashemi
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cell ◽  
Adhesion Molecule ◽  
Cell Adhesion Molecule ◽  
Extracellular Domain ◽  
Epithelial Cell Adhesion Molecule ◽  
Single Chain ◽  
E Coli ◽  
The Impact

Abstract Background Overexpression of the EpCAM (epithelial cell adhesion molecule) in malignancies makes it an attractive target for passive immunotherapy in a wide range of carcinomas. In comparison with full-length antibodies, due to the small size, the scFvs (single-chain variable fragments) are more suitable for recombinant expression in E. coli (Escherichia coli). However, the proteins expressed in large amounts in E. coli tend to form inclusion bodies that need to be refolded which may result in poor recovery of bioactive proteins. Various engineered strains were shown to be able to alleviate the insolubility problem. Here, we studied the impact of four E. coli strains on the soluble level of anti-EpEX-scFv (anti-EpCAM extracellular domain-scFv) protein. Results Although results showed that the amount of soluble anti-EpEX-scFv obtained in BL21TM (DE3) (114.22 ± 3.47 mg/L) was significantly higher to those produced in the same condition in E. coli RosettaTM (DE3) (71.39 ± 0.31 mg/L), and OrigamiTM T7 (58.99 ± 0.44 mg/L) strains, it was not significantly different from that produced by E. coli SHuffleTM T7 (108.87 ± 2.71 mg/L). Furthermore, the highest volumetric productivity of protein reached 318.29 ± 26.38 mg/L in BL21TM (DE3). Conclusions Although BL21TM (DE3) can be a suitable strain for high-level production of anti-EpEX-scFv protein, due to higher solubility yield (about 55%), E. coli SHuffleTM T7 seems to be better candidate for soluble production of scfv compared to BL21TM (DE3) (solubility yield of about 30%).

scholarly journals Role of Escherichia coli O157:H7 Virulence Factors in Colonization at the Bovine Terminal Rectal Mucosa

2006 ◽  
Vol 74 (8) ◽  
pp. 4685-4693 ◽  
Author(s):  
Haiqing Sheng ◽  
Ji Youn Lim ◽  
Hannah J. Knecht ◽  
Jie Li ◽  
Carolyn J. Hovde
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Clinical Manifestations ◽  
Rectal Mucosa ◽  
Wild Type ◽  
E Coli ◽  
Healthy Cattle ◽  
Life Threatening ◽  
K 12 ◽  
The Impact

ABSTRACT The human pathogen Escherichia coli O157:H7 causes hemorrhagic colitis and life-threatening sequelae and transiently colonizes healthy cattle at the terminal rectal mucosa. This study analyzed virulence factors important for the clinical manifestations of human E. coli O157:H7 infection for their contribution to the persistence of E. coli in cattle. The colonizing ability of E. coli O157:H7 was compared with those of nonpathogenic E. coli K-12 and isogenic deletion mutants missing Shiga toxin (Stx), the adhesin intimin, its receptor Tir, hemolysin, or the ∼92-kb pO157. Fully ruminant steers received a single rectal application of one E. coli strain so that effects of mucosal attachment and survival at the terminal rectum could be measured without the impact of bacterial passage through the entire gastrointestinal tract. Colonization was monitored by sensitive recto-anal junction mucosal swab culture. Nonpathogenic E. coli K-12 did not colonize as well as E. coli O157:H7 at the bovine terminal rectal mucosa. The E. coli O157:H7 best able to persist had intimin, Tir, and the pO157. Strains missing even one of these factors were recovered in lower numbers and were cleared faster than the wild type. In contrast, E. coli O157:H7 strains that were missing Stx or hemolysin colonized like the wild type. For these three strains, the number of bacteria increased between days 1 and 4 postapplication and then decreased slowly. In contrast, the numbers of noncolonizing strains (K-12, Δtir, and Δeae) decreased from the day of application. These patterns consistently predicted long-term colonization or clearance of the bacteria from the bovine terminal rectal mucosa.

scholarly journals Enterohemorrhagic Escherichia coli O157:H7 Disrupts Stat1-Mediated Gamma Interferon Signal Transduction in Epithelial Cells

2003 ◽  
Vol 71 (3) ◽  
pp. 1396-1404 ◽  
Author(s):  
Peter J. M. Ceponis ◽  
Derek M. McKay ◽  
Joyce C. Y. Ching ◽  
Perpetual Pereira ◽  
Philip M. Sherman
Keyword(s):  
Escherichia Coli ◽  
Signal Transduction ◽  
Epithelial Cells ◽  
Host Cell ◽  
Tyrosine Phosphorylation ◽  
Gamma Interferon ◽  
Enterohemorrhagic Escherichia Coli ◽  
Cell Protein ◽  
E Coli ◽  
Ifn Γ

ABSTRACT Enterohemorrhagic Escherichia coli (EHEC) O157:H7 is a clinically important bacterial enteropathogen that manipulates a variety of host cell signal transduction cascades to establish infection. However, the effect of EHEC O157:H7 on Jak/Stat signaling is unknown. To define the effect of EHEC infection on epithelial gamma interferon (IFN-γ)-Stat1 signaling, human T84 and HEp-2 epithelial cells were infected with EHEC O157:H7 and then stimulated with recombinant human IFN-γ. Cells were also infected with different EHEC strains, heat-killed EHEC, enteropathogenic E. coli (EPEC) O127:H6, and the commensal strain E. coli HB101. Nuclear and whole-cell protein extracts were prepared and were assayed by an electrophoretic mobility shift assay (EMSA) and by Western blotting, respectively. Cells were also processed for immunofluorescence to detect the subcellular localization of Stat1. The EMSA revealed inducible, but not constitutive, Stat1 activation upon IFN-γ treatment of both cell lines. The EMSA also showed that 6 h of EHEC O157:H7 infection, but not 30 min of EHEC O157:H7 infection, prevented subsequent Stat1 DNA binding induced by IFN-γ, whereas infection with EPEC did not. Immunoblotting showed that infection with EHEC, but not infection with EPEC, eliminated IFN-γ-induced Stat1 tyrosine phosphorylation in both dose- and time-dependent fashions and disrupted inducible protein expression of the Stat1-dependent gene interferon regulatory factor 1. Immunofluorescence revealed that EHEC infection did not prevent nuclear accumulation of Stat1 after IFN-γ treatment. Also, Stat1 tyrosine phosphorylation was suppressed by different EHEC isolates, including intimin-, type III secretion- and plasmid-deficient strains, but not by HB101 and heat-killed EHEC. These findings indicate the novel disruption of host cell signaling caused by EHEC infection but not by EPEC infection.

scholarly journals Escherichia coli DraE Adhesin-Associated Bacterial Internalization by Epithelial Cells Is Promoted Independently by Decay-Accelerating Factor and Carcinoembryonic Antigen-Related Cell Adhesion Molecule Binding and Does Not Require the DraD Invasin

2008 ◽  
Vol 76 (9) ◽  
pp. 3869-3880 ◽  
Author(s):  
Natalia Korotkova ◽  
Yuliya Yarova-Yarovaya ◽  
Veronika Tchesnokova ◽  
Nina Yazvenko ◽  
Mike A. Carl ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Epithelial Cells ◽  
Carcinoembryonic Antigen ◽  
Host Cells ◽  
Decay Accelerating Factor ◽  
E Coli ◽  
Related Cell ◽  
Bladder Cells ◽  
Tract Infections

ABSTRACT The Dr family of Escherichia coli adhesins are virulence factors associated with diarrhea and urinary tract infections. Dr fimbriae are comprised of two subunits. DraE/AfaE represents the major structural, antigenic, and adhesive subunit, which recognizes decay-accelerating factor (DAF) and carcinoembryonic antigen (CEA)-related cell adhesion molecules (CEACAMs) CEA, CEACAM1, CEACAM3, and CEACAM6 as binding receptors. The DraD/AfaD subunit caps fimbriae and has been implicated in the entry of Dr-fimbriated E. coli into host cells. In this study, we demonstrate that DAF or CEACAM receptors independently promote DraE-mediated internalization of E. coli by CHO cell transfectants expressing these receptors. We also found that DraE-positive recombinant bacteria adhere to and are internalized by primary human bladder epithelial cells which express DAF and CEACAMs. DraE-mediated bacterial internalization by bladder cells was inhibited by agents which disrupt lipid rafts, microtubules, and phosphatidylinositol 3-kinase (PI3K) activity. Immunofluorescence confocal microscopic examination of epithelial cells detected considerable recruitment of caveolin, β1 integrin, phosphorylated ezrin, phosphorylated PI3K, and tubulin, but not F-actin, by cell-associated bacteria. Finally, we demonstrate that the DraD subunit, previously implicated as an “invasin,” is not required for β1 integrin recruitment or bacterial internalization.

scholarly journals Virulence factors of Escherichia coli: an overview of animal and human infections with emphasis in bovine mastitis

2019 ◽  
Vol 40 (5) ◽  
pp. 2087
Author(s):  
Simony Trevizan Guerra ◽  
Carolina Lechinski de Paula ◽  
Carmen Alicia Daza Bolaños ◽  
Rodrigo Tavanelli Hernandes ◽  
Márcio Garcia Ribeiro
Keyword(s):  
Escherichia Coli ◽  
Virulence Factors ◽  
Bovine Mastitis ◽  
Clinical Mastitis ◽  
Clinical Severity ◽  
E Coli ◽  
Intestinal Infections ◽  
Wide Range ◽  
Environmental Agents ◽  
The Impact

Escherichia coli is a normal inhabitant of the enteric microflora of human and animal. Intestinal and extra-intestinal infections caused by E. coli in mammals are characterized by the presence of diversity of virulence factors. In addition it can be isolated from environment surrounding human and animal farms. E. coli is the main environmental pathogen causing clinical mastitis in dairy cattle. It causes a wide range of disease severity, from changes seen exclusively in milk to severe systemic signs. The severity of clinical mastitis has been conventionally classified into three levels: mild (grade 1), moderate (score 2), and severe (score 3). Recently, reports of cases of bovine mastitis caused by environmental agents have been on the rise, in particular in countries that have succeeded in controlling contagious microorganisms. Unlike enteric and certain extra-enteric conditions in domestic animals and humans, the impact of virulence factors on the occurrence of bovine mastitis due to E. coli, as well as the clinical severity of the cases, is not fully understood. In this regard, the present study reviewed the most relevant virulence factors of E. coli in human and animals, with emphasis in bovine mastitis.

scholarly journals Molecular Characterization of the Acid-Inducible asr Gene of Escherichia coli and Its Role in Acid Stress Response

2003 ◽  
Vol 185 (8) ◽  
pp. 2475-2484 ◽  
Author(s):  
Vaida Šeputienė ◽  
Domantas Motiejūnas ◽  
Kęstutis Sužiedėlis ◽  
Henrik Tomenius ◽  
Staffan Normark ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Amino Acids ◽  
Signal Peptide ◽  
Polyacrylamide Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Acid Stress ◽  
Acid Tolerance ◽  
Site Directed Mutagenesis ◽  
E Coli ◽  
Acid Shock

ABSTRACT Enterobacteria have developed numerous constitutive and inducible strategies to sense and adapt to an external acidity. These molecular responses require dozens of specific acid shock proteins (ASPs), as shown by genomic and proteomic analysis. Most of the ASPs remain poorly characterized, and their role in the acid response and survival is unknown. We recently identified an Escherichia coli gene, asr (acid shock RNA), encoding a protein of unknown function, which is strongly induced by high environmental acidity (pH < 5.0). We show here that Asr is required for growth at moderate acidity (pH 4.5) as well as for the induction of acid tolerance at moderate acidity, as shown by its ability to survive subsequent transfer to extreme acidity (pH 2.0). Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western analysis of acid-shocked E. coli cells harboring a plasmid-borne asr gene demonstrated that the Asr protein is synthesized as a precursor with an apparent molecular mass of 18 kDa. Mutational studies of the asr gene also demonstrated the Asr preprotein contains 102 amino acids. This protein is subjected to an N-terminal cleavage of the signal peptide and a second processing event, yielding 15- and 8-kDa products, respectively. Only the 8-kDa polypeptide was detected in acid-shocked cells containing only the chromosomal copy of the asr gene. N-terminal sequencing and site-directed mutagenesis revealed the two processing sites in the Asr protein precursor. Deletion of amino acids encompassing the processing site required for release of the 8-kDa protein resulted in an acid-sensitive phenotype similar to that observed for the asr null mutant, suggesting that the 8-kDa product plays an important role in the adaptation to acid shock. Analysis of Asr:PhoA fusions demonstrated a periplasmic location for the Asr protein after removal of the signal peptide. Homologues of the asr gene from other Enterobacteriaceae were cloned and shown to be induced in E. coli under acid shock conditions.

Acid Stress, Starvation, and Cold Stress Affect Poststress Behavior of Escherichia coli O157:H7 and Nonpathogenic Escherichia coli†

2001 ◽  
Vol 64 (7) ◽  
pp. 970-974 ◽  
Author(s):  
B. LEENANON ◽  
M. A. DRAKE
Keyword(s):  
Escherichia Coli ◽  
Heat Resistance ◽  
Cold Stress ◽  
Acid Stress ◽  
Escherichia Coli O157 ◽  
Acid Adaptation ◽  
E Coli ◽  
Freeze Thaw ◽  
Acid Shock ◽  
Rpos Mutant

The effects of acid shock, acid adaptation, starvation, and cold stress of Escherichia coli O157:H7 (ATCC 43895), an rpo S mutant (FRIK 816-3), and nonpathogenic E. coli (ATCC 25922) on poststress heat resistance and freeze–thaw resistance were investigated. Following stress, heat tolerance at 56°C and freeze–thaw resistance at −20 to 21°C were determined. Heat and freeze–thaw resistance of E. coli O157:H7 and nonpathogenic E. coli was enhanced after acid adaptation and starvation. Following cold stress, heat resistance of E. coli O157:H7 and nonpathogenic E. coli was decreased, while freeze–thaw resistance was increased. Heat and freeze–thaw resistance of the rpoS mutant was enhanced only after acid adaptation. Increased or decreased tolerance of acid-adapted, starved, or cold-stressed E. coli O157:H7 cells to heat or freeze–thaw processes should be considered when processing minimally processed or extended shelf-life foods.

scholarly journals Porcine and Bovine Forms of Lactoferrin Inhibit Growth of Porcine Enterotoxigenic Escherichia coli and Degrade Its Virulence Factors

2020 ◽  
Vol 86 (24) ◽  
Author(s):  
Matthias Dierick ◽  
Hans Van der Weken ◽  
Joanna Rybarczyk ◽  
Daisy Vanrompay ◽  
Bert Devriendt ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Epithelial Cells ◽  
Virulence Factors ◽  
Intestinal Epithelial Cells ◽  
Enterotoxigenic Escherichia Coli ◽  
Bovine Lactoferrin ◽  
Adhesion Assay ◽  
Intestinal Epithelial ◽  
Content Type ◽  
E Coli

ABSTRACT Postweaning diarrhea (PWD) is an economically important, multifactorial disease affecting pigs within the first 2 weeks after weaning. The most common agent associated with PWD is enterotoxigenic Escherichia coli (ETEC). Currently, antibiotics are used to control PWD, and this has most likely contributed to an increased prevalence of antibiotic-resistant strains. This puts pressure on veterinarians and farmers to decrease or even abandon the use of antibiotics, but these measures need to be supported by alternative strategies for controlling these infections. Naturally derived molecules, such as lactoferrin, could be potential candidates due to their antibacterial or immune-modulating activities. Here, we analyzed the ability of bovine lactoferrin (bLF), porcine lactoferrin (pLF), and ovotransferrin (ovoTF) to inhibit ETEC growth, degrade ETEC virulence factors, and inhibit adherence of these pathogens to porcine intestinal epithelial cells. Our results revealed that bLF and pLF, but not ovoTF, inhibit the growth of ETEC. Furthermore, bLF and pLF can degrade several virulence factors produced by ETEC strains, more specifically F4 fimbriae, F18 fimbriae, and flagellin. On the other hand, ovoTF degrades F18 fimbriae and flagellin but not F4 fimbriae. An in vitro adhesion assay showed that bLF, ovoTF, and pLF can decrease the number of bacteria adherent to epithelial cells. Our findings demonstrate that lactoferrin can directly affect porcine ETEC strains, which could allow lactoferrin to serve as an alternative to antimicrobials for the prevention of ETEC infections in piglets. IMPORTANCE Currently, postweaning F4+ and F18+ Escherichia coli infections in piglets are controlled by the use of antibiotics and zinc oxide, but the use of these antimicrobial agents most likely contributes to an increase in antibiotic resistance. Our work demonstrates that bovine and porcine lactoferrin can inhibit the growth of porcine enterotoxigenic E. coli strains. In addition, we also show that lactoferrin can reduce the adherence of these strains to small intestinal epithelial cells, even at a concentration that does not inhibit bacterial growth. This research could allow us to develop lactoferrin as an alternative strategy to prevent enterotoxigenic E. coli (ETEC) infections in piglets.

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