Antigen 43 from Escherichia coli Induces Inter- and Intraspecies Cell Aggregation and Changes in Colony Morphology of Pseudomonas fluorescens

ABSTRACT Antigen 43 (Ag43) is a surface-displayed autotransporter protein ofEscherichia coli. By virtue of its self-association characteristics, this protein is able to mediate autoaggregation and flocculation of E. coli cells in static cultures. Additionally, surface display of Ag43 is associated with a distinct frizzy colony morphology in E. coli. Here we show that Ag43 can be expressed in a functional form on the surface of the environmentally important Pseudomonas fluorescens strain SBW25 with ensuing cell aggregation and frizzy colony types. Using green fluorescence protein-tagged cells, we demonstrate that Ag43 can be used as a tool to provide interspecies cell aggregation betweenE. coli and P. fluorescens. Furthermore, Ag43 expression enhances biofilm formation in P. fluorescens to glass surfaces. The versatility of this protein was also reflected in Ag43 surface display in a variety of other gram-negative bacteria. Display of heterologous Ag43 in selected bacteria might offer opportunities for rational design of multispecies consortia where the concerted action of several bacterial species is required, e.g., waste treatment and degradation of pollutants.

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Functional Analysis of Antigen 43 in Uropathogenic Escherichia coli Reveals a Role in Long-Term Persistence in the Urinary Tract

Infection and Immunity ◽

10.1128/iai.01952-06 ◽

2007 ◽

Vol 75 (7) ◽

pp. 3233-3244 ◽

Cited By ~ 120

Author(s):

Glen C. Ulett ◽

Jaione Valle ◽

Christophe Beloin ◽

Orla Sherlock ◽

Jean-Marc Ghigo ◽

...

Keyword(s):

Escherichia Coli ◽

Urinary Tract ◽

Biological Significance ◽

Cell Aggregation ◽

Upec Strain ◽

Biofilm Growth ◽

E Coli ◽

Antigen 43 ◽

K 12

ABSTRACT Escherichia coli is the primary cause of urinary tract infection (UTI) in the developed world. The major factors associated with the virulence of uropathogenic E. coli (UPEC) are fimbrial adhesins, which mediate specific attachment to host receptors and trigger innate host responses. Another group of adhesins is represented by the autotransporter subgroup of proteins. The best characterized of these proteins, antigen 43 (Ag43), is a self-recognizing adhesin that is associated with cell aggregation and biofilm formation in E. coli K-12. The sequenced genome of prototype UPEC strain CFT073 contains two variant Ag43-encoding genes located on pathogenicity islands. The biological significance of both of these genes and their role in UPEC pathogenesis have not been investigated previously. Here we performed a detailed molecular characterization analysis of Ag43a (c3655) and Ag43b (c1273) from UPEC CFT073. Expression of Ag43a and Ag43b in a K-12 background revealed that they possess different functional properties. Ag43a produced a strong aggregation phenotype and promoted significant biofilm growth. Deletion mutants and strains constitutively expressing Ag43a and Ag43b were also constructed using CFT073. When these mutants were analyzed in a mouse model of UTI, Ag43a (but not Ag43b) promoted long-term persistence in the urinary bladder. Our findings demonstrate that Ag43a contributes to UPEC disease pathogenesis and reveal that there are pathogenicity-adapted variants of Ag43 with distinct virulence-related functions.

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Characterization of Escherichia coli Strains Derived from Cow Milk of Subclinical and Clinical Cases of Mastitis

Applied Sciences ◽

10.3390/app11020541 ◽

2021 ◽

Vol 11 (2) ◽

pp. 541

Author(s):

Katarzyna Grudlewska-Buda ◽

Krzysztof Skowron ◽

Ewa Wałecka-Zacharska ◽

Natalia Wiktorczyk-Kapischke ◽

Jarosław Bystroń ◽

...

Keyword(s):

Escherichia Coli ◽

Bacterial Species ◽

Subclinical Mastitis ◽

Clinical Mastitis ◽

Economic Problem ◽

Cow Milk ◽

P Value ◽

Dairy Herds ◽

E Coli ◽

Bonferroni Test

Mastitis is a major economic problem in dairy herds, as it might decrease fertility, and negatively affect milk quality and milk yield. Out of over 150 bacterial species responsible for the udder inflammation, Escherichia coli is one of the most notable. This study aimed to assess antimicrobial susceptibility, resistance to dipping agents and biofilm formation of 150 E. coli strains isolated from milk of cows with subclinical and clinical mastitis. The strains came from three dairy herds located in Northern and Central Poland. The statistical analyses were performed with post-hoc Bonferroni test and chi-square test (including Yates correction). The data with a p value of <0.05 were considered significant. We found that the tested strains were mostly sensitive to antimicrobials and dipping agents. It was shown that 37.33% and 4.67% of strains were resistant and moderately resistant to at least one antimicrobial agent, respectively. No extended-spectrum beta-lactamases (ESBL)-producing E. coli were detected. The majority of strains did not possess the ability to form biofilm or formed a weak biofilm. The strong biofilm formers were found only among strains derived from cows with subclinical mastitis. The lowest bacteria number was noted for subclinical mastitis cows’ strains, after stabilization with iodine (3.77 log CFU × cm−2) and chlorhexidine (3.96 log CFU × cm−2) treatment. In the present study, no statistically significant differences in susceptibility to antibiotics and the ability to form biofilm were found among the strains isolated from cows with subclinical and clinical mastitis. Despite this, infections in dairy herds should be monitored. Limiting the spread of bacteria and characterizing the most common etiological factors would allow proper treatment.

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Toxicity of the First Ten MEIC Chemicals to Bacteria

Alternatives to Laboratory Animals ◽

10.1177/026119299302100205 ◽

1993 ◽

Vol 21 (2) ◽

pp. 151-155

Author(s):

Gustaw Kerszman

Keyword(s):

Escherichia Coli ◽

Bacillus Subtilis ◽

Linear Regression ◽

Minimal Inhibitory Concentration ◽

Blood Concentration ◽

Inhibitory Concentration ◽

Human Lymphocytes ◽

Bacterial Species ◽

E Coli ◽

Mild Toxicity

The toxicity of the first ten MEIC chemicals to Escherichia coli and Bacillus subtilis was examined. Nine of the chemicals were toxic to the bacteria, with the minimal inhibitory concentration (MIC) ranging from 10-3 to 4.4M. The sensitivities of both organisms were similar, but the effect on E. coli was often bactericidal, while it was bacteriostatic for B. subtilis. Digoxin was not detectably toxic to either bacterial species. Amitriptyline and FeSO4 were relatively less toxic to the bacteria than to human cells. For seven chemicals, a highly significant linear regression was established between log MIC in bacteria and log of blood concentration, giving lethal and moderate/mild toxicity in humans, as well as with toxicity to human lymphocytes.

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Preparation of Sticky Escherichia coli through Surface Display of an Adhesive Catecholamine Moiety

Applied and Environmental Microbiology ◽

10.1128/aem.02223-13 ◽

2013 ◽

Vol 80 (1) ◽

pp. 43-53 ◽

Cited By ~ 18

Author(s):

Joseph P. Park ◽

Min-Jung Choi ◽

Se Hun Kim ◽

Seung Hwan Lee ◽

Haeshin Lee

Keyword(s):

Escherichia Coli ◽

Cell Surface ◽

Communication Systems ◽

Cell Attachment ◽

Surface Display ◽

Content Type ◽

E Coli ◽

Material Surfaces ◽

Mussel Adhesive ◽

Adhesive Proteins

ABSTRACTMussels attach to virtually all types of inorganic and organic surfaces in aqueous environments, and catecholamines composed of 3,4-dihydroxy-l-phenylalanine (DOPA), lysine, and histidine in mussel adhesive proteins play a key role in the robust adhesion. DOPA is an unusual catecholic amino acid, and its side chain is called catechol. In this study, we displayed the adhesive moiety of DOPA-histidine onEscherichia colisurfaces using outer membrane protein W as an anchoring motif for the first time. Localization of catecholamines on the cell surface was confirmed by Western blot and immunofluorescence microscopy. Furthermore, cell-to-cell cohesion (i.e., cellular aggregation) induced by the displayed catecholamine and synthesis of gold nanoparticles on the cell surface support functional display of adhesive catecholamines. The engineeredE. coliexhibited significant adhesion onto various material surfaces, including silica and glass microparticles, gold, titanium, silicon, poly(ethylene terephthalate), poly(urethane), and poly(dimethylsiloxane). The uniqueness of this approach utilizing the engineered stickyE. coliis that no chemistry for cell attachment are necessary, and the ability of spontaneousE. coliattachment allows one to immobilize the cells on challenging material surfaces such as synthetic polymers. Therefore, we envision that mussel-inspired catecholamine yielded stickyE. colithat can be used as a new type of engineered microbe for various emerging fields, such as whole living cell attachment on versatile material surfaces, cell-to-cell communication systems, and many others.

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Involvement of the cgtA gene function in stimulation of DNA repair in Escherichia coli and Vibrio harveyi

Microbiology ◽

10.1099/mic.0.26292-0 ◽

2003 ◽

Vol 149 (7) ◽

pp. 1763-1770 ◽

Cited By ~ 12

Author(s):

Ryszard Zielke ◽

Aleksandra Sikora ◽

Rafał Dutkiewicz ◽

Grzegorz Wegrzyn ◽

Agata Czyż

Keyword(s):

Gene Expression ◽

Escherichia Coli ◽

Dna Repair ◽

Gene Product ◽

Uv Irradiation ◽

Vibrio Harveyi ◽

Bacterial Species ◽

Wild Type ◽

E Coli ◽

Stimulation Of

CgtA is a member of the Obg/Gtp1 subfamily of small GTP-binding proteins. CgtA homologues have been found in various prokaryotic and eukaryotic organisms, ranging from bacteria to humans. Nevertheless, despite the fact that cgtA is an essential gene in most bacterial species, its function in the regulation of cellular processes is largely unknown. Here it has been demonstrated that in two bacterial species, Escherichia coli and Vibrio harveyi, the cgtA gene product enhances survival of cells after UV irradiation. Expression of the cgtA gene was found to be enhanced after UV irradiation of both E. coli and V. harveyi. Moderate overexpression of cgtA resulted in higher UV resistance of E. coli wild-type and dnaQ strains, but not in uvrA, uvrB, umuC and recA mutant hosts. Overexpression of the E. coli recA gene in the V. harveyi cgtA mutant, which is very sensitive to UV light, restored the level of survival of UV-irradiated cells to the levels observed for wild-type bacteria. Moreover, the basal level of the RecA protein was lower in a temperature-sensitive cgtA mutant of E. coli than in the cgtA + strain, and contrary to wild-type bacteria, no significant increase in recA gene expression was observed after UV irradiation of this cgtA mutant. Finally, stimulation of uvrB gene transcription under these conditions was impaired in the V. harveyi cgtA mutant. All these results strongly suggest that the cgtA gene product is involved in DNA repair processes, most probably by stimulation of recA gene expression and resultant activation of RecA-dependent DNA repair pathways.

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High Resolution Melt Assays to Detect and Identify Vibrio parahaemolyticus, Bacillus cereus, Escherichia coli, and Clostridioides difficile Bacteria

Microorganisms ◽

10.3390/microorganisms8040561 ◽

2020 ◽

Vol 8 (4) ◽

pp. 561

Author(s):

Allison C. Bender ◽

Jessica A. Faulkner ◽

Katherine Tulimieri ◽

Thomas H. Boise ◽

Kelly M. Elkins

Keyword(s):

Escherichia Coli ◽

High Resolution ◽

Bacillus Cereus ◽

Vibrio Parahaemolyticus ◽

Bacterial Species ◽

Human Pathogens ◽

High Resolution Melt ◽

E Coli ◽

Clostridioides Difficile ◽

Sensitivity Specificity

Over one hundred bacterial species have been determined to comprise the human microbiota in a healthy individual. Bacteria including Escherichia coli, Bacillus cereus, Clostridioides difficile, and Vibrio parahaemolyticus are found inside of the human body and B. cereus and E. coli are also found on the skin. These bacteria can act as human pathogens upon ingestion of contaminated food or water, if they enter an open wound, or antibiotics, and environment or stress can alter the microbiome. In this study, we present new polymerase chain reaction (PCR) high-resolution melt (HRM) assays to detect and identify the above microorganisms. Amplified DNA from C. difficile, E. coli, B. cereus, and V. parahaemolyticus melted at 80.37 ± 0.45 °C, 82.15 ± 0.37 °C, 84.43 ± 0.50 °C, and 86.74 ± 0.65 °C, respectively. A triplex PCR assay was developed to simultaneously detect and identify E. coli, B. cereus, and V. parahaemolyticus, and cultured microorganisms were successfully amplified, detected, and identified. The assays demonstrated sensitivity, specificity, reproducibility, and robustness in testing.

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Inactivation of Multi-Drug Resistant Non-Typhoidal Salmonella and Wild-Type Escherichia coli STEC Using Organic Acids: A Potential Alternative to the Food Industry

Pathogens ◽

10.3390/pathogens9100849 ◽

2020 ◽

Vol 9 (10) ◽

pp. 849

Author(s):

Vinicius Silva Castro ◽

Yhan da Silva Mutz ◽

Denes Kaic Alves Rosario ◽

Adelino Cunha-Neto ◽

Eduardo Eustáquio de Souza Figueiredo ◽

...

Keyword(s):

Escherichia Coli ◽

Acetic Acid ◽

Organic Acids ◽

Sodium Hypochlorite ◽

Bacterial Species ◽

Disk Diffusion ◽

Clinical Strains ◽

Linear Pattern ◽

E Coli ◽

Log Reduction

Salmonella and Escherichia coli are the main bacterial species involved in food outbreaks worldwide. Recent reports showed that chemical sanitizers commonly used to control these pathogens could induce antibiotic resistance. Therefore, this study aimed to describe the efficiency of chemical sanitizers and organic acids when inactivating wild and clinical strains of Salmonella and E. coli, targeting a 4-log reduction. To achieve this goal, three methods were applied. (i) Disk-diffusion challenge for organic acids. (ii) Determination of MIC for two acids (acetic and lactic), as well as two sanitizers (quaternary compound and sodium hypochlorite). (iii) The development of inactivation models from the previously defined concentrations. In disk-diffusion, the results indicated that wild strains have higher resistance potential when compared to clinical strains. Regarding the models, quaternary ammonium and lactic acid showed a linear pattern of inactivation, while sodium hypochlorite had a linear pattern with tail dispersion, and acetic acid has Weibull dispersion to E. coli. The concentration to 4-log reduction differed from Salmonella and E. coli in acetic acid and sodium hypochlorite. The use of organic acids is an alternative method for antimicrobial control. Our study indicates the levels of organic acids and sanitizers to be used in the inactivation of emerging foodborne pathogens.

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Evidence for Transfer of CMY-2 AmpC β-Lactamase Plasmids between Escherichia coli andSalmonella Isolates from Food Animals and Humans

Antimicrobial Agents and Chemotherapy ◽

10.1128/aac.45.10.2716-2722.2001 ◽

2001 ◽

Vol 45 (10) ◽

pp. 2716-2722 ◽

Cited By ~ 249

Author(s):

P. L. Winokur ◽

D. L. Vonstein ◽

L. J. Hoffman ◽

E. K. Uhlenhopp ◽

G. V. Doern

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Bacterial Species ◽

Laboratory Strain ◽

Epidemiological Studies ◽

Geographic Region ◽

Chromosomal Dna ◽

E Coli ◽

Clonal Relatedness ◽

Food Animals

ABSTRACT Escherichia coli is an important pathogen that shows increasing antimicrobial resistance in isolates from both animals and humans. Our laboratory recently described Salmonellaisolates from food animals and humans that expressed an identical plasmid-mediated, AmpC-like β-lactamase, CMY-2. In the present study, 59 of 377 E. coli isolates from cattle and swine (15.6%) and 6 of 1,017 (0.6%) isolates of human E. coli from the same geographic region were resistant to both cephamycins and extended-spectrum cephalosporins. AnampC gene could be amplified with CMY-2 primers in 94.8% of animal and 33% of human isolates. Molecular epidemiological studies of chromosomal DNA revealed little clonal relatedness among the animal and human E. coli isolates harboring the CMY-2 gene. The ampC genes from 10 animal and human E. coli isolates were sequenced, and all carried an identical CMY-2 gene. Additionally, all were able to transfer a plasmid containing the CMY-2 gene to a laboratory strain of E. coli. CMY-2 plasmids demonstrated two different plasmid patterns that each showed strong similarities to previously describedSalmonella CMY-2 plasmids. Additionally, Southern blot analyses using a CMY-2 probe demonstrated conserved fragments among many of the CMY-2 plasmids identified in Salmonella andE. coli isolates from food animals and humans. These data demonstrate that common plasmids have been transferred between animal-associated Salmonella and E. coli, and identical CMY-2 genes carried by similar plasmids have been identified in humans, suggesting that the CMY-2 plasmid has undergone transfer between different bacterial species and may have been transmitted between food animals and humans.

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Novel Roles for the AIDA Adhesin from Diarrheagenic Escherichia coli: Cell Aggregation and Biofilm Formation

Journal of Bacteriology ◽

10.1128/jb.186.23.8058-8065.2004 ◽

2004 ◽

Vol 186 (23) ◽

pp. 8058-8065 ◽

Cited By ~ 128

Author(s):

Orla Sherlock ◽

Mark A. Schembri ◽

Andreas Reisner ◽

Per Klemm

Keyword(s):

Escherichia Coli ◽

Biofilm Formation ◽

Mammalian Cells ◽

Gastrointestinal Disease ◽

Cell Aggregation ◽

Bacterial Attachment ◽

E Coli ◽

Abiotic Surfaces ◽

Broad Variety ◽

Self Association

ABSTRACT Diarrhea-causing Escherichia coli strains are responsible for numerous cases of gastrointestinal disease and constitute a serious health problem throughout the world. The ability to recognize and attach to host intestinal surfaces is an essential step in the pathogenesis of such strains. AIDA is a potent bacterial adhesin associated with some diarrheagenic E. coli strains. AIDA mediates bacterial attachment to a broad variety of human and other mammalian cells. It is a surface-displayed autotransporter protein and belongs to the selected group of bacterial glycoproteins; only the glycosylated form binds to mammalian cells. Here, we show that AIDA possesses self-association characteristics and can mediate autoaggregation of E. coli cells. We demonstrate that intercellular AIDA-AIDA interaction is responsible for bacterial autoaggregation. Interestingly, AIDA-expressing cells can interact with antigen 43 (Ag43)-expressing cells, which is indicative of an intercellular AIDA-Ag43 interaction. Additionally, AIDA expression dramatically enhances biofilm formation by E. coli on abiotic surfaces in flow chambers.

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Adaptation in a Mouse Colony Monoassociated with Escherichia coli K-12 for More than 1,000 Days

Applied and Environmental Microbiology ◽

10.1128/aem.00358-10 ◽

2010 ◽

Vol 76 (14) ◽

pp. 4655-4663 ◽

Cited By ~ 13

Author(s):

Sean M. Lee ◽

Aaron Wyse ◽

Aaron Lesher ◽

Mary Lou Everett ◽

Linda Lou ◽

...

Keyword(s):

Escherichia Coli ◽

Growth Rates ◽

Bacterial Species ◽

Intestinal Flora ◽

Average Density ◽

Host Bacterium ◽

E Coli ◽

K 12

ABSTRACT Although mice associated with a single bacterial species have been used to provide a simple model for analysis of host-bacteria relationships, bacteria have been shown to display adaptability when grown in a variety of novel environments. In this study, changes associated with the host-bacterium relationship in mice monoassociated with Escherichia coli K-12 over a period of 1,031 days were evaluated. After 80 days, phenotypic diversification of E. coli was observed, with the colonizing bacteria having a broader distribution of growth rates in the laboratory than the parent E. coli. After 1,031 days, which included three generations of mice and an estimated 20,000 generations of E. coli, the initially homogeneous bacteria colonizing the mice had evolved to have widely different growth rates on agar, a potential decrease in tendency for spontaneous lysis in vivo, and an increased tendency for spontaneous lysis in vitro. Importantly, mice at the end of the experiment were colonized at an average density of bacteria that was more than 3-fold greater than mice colonized on day 80. Evaluation of selected isolates on day 1,031 revealed unique restriction endonuclease patterns and differences between isolates in expression of more than 10% of the proteins identified by two-dimensional electrophoresis, suggesting complex changes underlying the evolution of diversity during the experiment. These results suggest that monoassociated mice might be used as a tool for characterizing niches occupied by the intestinal flora and potentially as a method of targeting the evolution of bacteria for applications in biotechnology.

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