scholarly journals The ferric yersiniabactin uptake receptor FyuA is required for efficient biofilm formation by urinary tract infectious Escherichia coli in human urine

Microbiology ◽  
2008 ◽  
Vol 154 (1) ◽  
pp. 167-175 ◽  
Author(s):  
Viktoria Hancock ◽  
Lionel Ferrières ◽  
Per Klemm
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Human Urine ◽  
Biofilm Formation

scholarly journals Global Gene Expression Profiling of Asymptomatic Bacteriuria Escherichia coli during Biofilm Growth in Human Urine

2006 ◽  
Vol 75 (2) ◽  
pp. 966-976 ◽  
Author(s):  
Viktoria Hancock ◽  
Per Klemm
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Expression Profile ◽  
Human Urine ◽  
Biofilm Formation ◽  
Expression Profiles ◽  
Asymptomatic Bacteriuria ◽  
Knockout Mutant ◽  
Biofilm Growth ◽  
E Coli

ABSTRACT Urinary tract infection (UTI) is an important health problem worldwide, with many millions of cases each year, and Escherichia coli is the most common organism causing UTI in humans. Also, E. coli is responsible for most infections in patients with chronic indwelling bladder catheter. The two asymptomatic bacteriuria (ABU) E. coli strains 83972 and VR50 are significantly better biofilm formers in their natural growth medium, human urine, than the two uropathogenic E. coli isolates CFT073 and 536. We used DNA microarrays to monitor the expression profile during biofilm growth in urine of the two ABU strains 83972 and VR50. Significant differences in expression levels were seen between the biofilm expression profiles of the two strains with the corresponding planktonic expression profiles in morpholinepropanesulfonic acid minimal laboratory medium and human urine; 417 and 355 genes were up- and down-regulated, respectively, during biofilm growth in urine of 83972 and VR50. Many genes involved in transcription and stress were up-regulated in biofilm-grown cells. The role in biofilm formation of four of the up-regulated genes, i.e., yceP, yqgA, ygiD, and aaeX, was investigated by creating single-knockout mutant versions of 83972 and VR50; all mutants showed reduced biofilm formation in urine by 18 to 43% compared with the wild type (P < 0.05). Also, the expression profile of strain 83972 in the human urinary tract partially overlaps with the biofilm expression profile.

scholarly journals Distribution of genes encoding adhesins and biofilm formation capacity among Uropathogenic Escherichia coli isolates in relation to the antimicrobial resistance

2020 ◽  
Vol 20 (1) ◽  
pp. 238-247
Author(s):  
Ashraf A Kadry ◽  
Nour M Al-Kashef ◽  
Amira M El-Ganiny
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Antimicrobial Resistance ◽  
Biofilm Formation ◽  
Critical Role ◽  
Uropathogenic Escherichia Coli ◽  
Diffusion Method ◽  
Esbl Production ◽  
Microtitre Plate Assay ◽  
Genes Encoding

Background: Escherichia coli is the most predominant pathogen involved in UTIs. Mainly, fimbrial surface appendages are impli- cated in adherence to urothelium besides non-fimbrial proteins. Objectives: To determine prevalence of genes encoding fimbrial and non-fimbrial proteins among Uropathogenic Escherichia coli (UPEC). Furthermore, distribution of these genes and biofilm formation capacity were investigated in relation to antimicrobial resistance. Methods: Antimicrobial susceptibility of 112 UPEC isolates was performed using disc diffusion method. ESBL production was confirmed by double disc synergy test. Genes encoding fimbrial and non-fimbrial proteins were detected using PCR and biofilm formation was investigated using microtitre plate assay. Results: UPEC isolates exhibited high resistance against doxycyclines (88.39 %), β-lactams (7.14-86.6%), sulphamethoxaz- ole–trimethoprim (53.75%) and fluoro-quinolones (50%). Fifty percent of tested isolates were ESBL producers. PapGII gene was statistically more prevalent among pyelonephritis isolates. SfaS, focG and picU genes were statistically associated with flu- oro-quinolone (FQs) sensitive isolates and Dr/afaBC gene was statistically associated with ESBL production. Moreover, non- MDR isolates produced sturdier biofilm. Conclusion: PapGII adhesin variant seems to have a critical role in colonization of upper urinary tract. There is a possible link between antimicrobial resistance and virulence being capable of affecting the distribution of some genes besides its negative impact on biofilm formation. Keywords: Urinary tract infection; Escherichia coli; UPEC; adhesin genes; ESBL; biofilm.  

scholarly journals Implication of biofilm formation in the persistence of urinary tract infection caused by uropathogenic Escherichia coli

2006 ◽  
Vol 12 (10) ◽  
pp. 1034-1036 ◽  
Author(s):  
S.M. Soto ◽  
A. Smithson ◽  
J.P. Horcajada ◽  
J.A. Martinez ◽  
J.P. Mensa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Biofilm Formation ◽  
Uropathogenic Escherichia Coli

scholarly journals PhoU enhances the ability of extraintestinal pathogenic Escherichia coli strain CFT073 to colonize the murine urinary tract

Microbiology ◽  
2006 ◽  
Vol 152 (1) ◽  
pp. 153-160 ◽  
Author(s):  
Eric L. Buckles ◽  
Xiaolin Wang ◽  
C. Virginia Lockatell ◽  
David E. Johnson ◽  
Michael S. Donnenberg
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract Infection ◽  
Urinary Tract ◽  
Tract Infection ◽  
Mutant Strain ◽  
Human Urine ◽  
Type Strain ◽  
Wild Type Strain ◽  
Wild Type ◽  
Strain Cft073

The phoU gene is the last cistron in the pstSCAB–phoU operon and functions as a negative regulator of the Pho regulon. The authors previously identified a phoU mutant of extraintestinal pathogenic Escherichia coli strain CFT073 and demonstrated that this mutant was attenuated for survival in the murine model of ascending urinary tract infection. It is hypothesized that the PhoU protein might serve as a urovirulence factor by indirectly affecting the expression of virulence-related genes. In this study, the phoU mutant was further characterized and PhoU was confirmed as a virulence factor. Western blot analysis demonstrated that insertion of the transposon in the phoU gene disrupted the expression of PhoU. The phoU mutant had derepressed alkaline phosphatase activity under phosphate-excess and -limiting conditions. In single-challenge murine ascending urinary tract infection experiments, quantitative cultures of urine, bladder and kidney revealed no significant differences between the phoU mutant strain and the wild-type strain CFT073. However, in competitive colonization experiments, the phoU mutant strain was significantly out-competed by the wild-type strain in the kidneys and urine and recovered in lower amount in the bladder. Complementation of the phoU mutant with a plasmid containing the wild-type phoU gene restored the expression of PhoU and alkaline phosphate activity to wild-type levels and no significant difference in colonization was observed between the phoU mutant containing the complementing plasmid and wild-type in competitive colonization experiments. In human urine, the phoU mutant and wild-type grew comparably when inoculated independently, indicating that the attenuation observed was not due to a general growth defect. However, as observed in vivo, the wild-type out-competed the phoU mutant in competition growth experiments in human urine. These data indicate that PhoU contributes to efficient colonization of the murine urinary tract and add PhoU to a short list of confirmed urovirulence factors.

scholarly journals Abolition of Biofilm Formation in Urinary Tract Escherichia coli and Klebsiella Isolates by Metal Interference through Competition for Fur

2010 ◽  
Vol 76 (12) ◽  
pp. 3836-3841 ◽  
Author(s):  
Viktoria Hancock ◽  
Malin Dahl ◽  
Per Klemm
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Biofilm Formation ◽  
Iron Uptake ◽  
Regulatory System ◽  
Bacterial Biofilms ◽  
Divalent Metal Ions ◽  
Chronic Infections ◽  
E Coli ◽  
Microtiter Plates

ABSTRACT Bacterial biofilms are associated with a large number of persistent and chronic infections. Biofilm-dwelling bacteria are particularly resistant to antibiotics and immune defenses, which makes it hard if not impossible to eradicate biofilm-associated infections. In the urinary tract, free iron is strictly limited but is critical for bacterial growth. Biofilm-associated Escherichia coli cells are particularly desperate for iron. An attractive way of inhibiting biofilm formation is to fool the bacterial regulatory system for iron uptake. Here, we demonstrate that biofilm formation can be impaired by the addition of divalent metal ions, such as Zn(II) and Co(II), which inhibit iron uptake by virtue of their higher-than-iron affinity for the master controller protein of iron uptake, Fur. Reduced biofilm formation of urinary tract-infectious E. coli strains in the presence of Zn(II) was observed in microtiter plates and flow chambers as well as on urinary catheters. These results further support that iron uptake is indeed crucial for biofilm formation, and thereby, targeting these uptake systems might be an effective way to eradicate biofilms caused by infectious strains.

Chlorpromazine-impregnated catheters as a potential strategy to control biofilm-associated urinary tract infections

2019 ◽  
Vol 14 (12) ◽  
pp. 1023-1034 ◽  
Author(s):  
José JC Sidrim ◽  
Bruno R Amando ◽  
Francisco IF Gomes ◽  
Marilia SMG do Amaral ◽  
Paulo CP de Sousa ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Klebsiella Pneumoniae ◽  
Urinary Tract Infections ◽  
Proteus Mirabilis ◽  
Biofilm Formation ◽  
Antibiofilm Activity ◽  
Potential Strategy ◽  
Tract Infections

Aim: This study proposes the impregnation of Foley catheters with chlorpromazine (CPZ) to control biofilm formation by Escherichia coli, Proteus mirabilis and Klebsiella pneumoniae. Materials & methods: The minimum inhibitory concentrations (MICs) for CPZ and the effect of CPZ on biofilm formation were assessed. Afterward, biofilm formation and the effect of ciprofloxacin and meropenem (at MIC) on mature biofilms grown on CPZ-impregnated catheters were evaluated. Results: CPZ MIC range was 39.06–625 mg/l. CPZ significantly reduced (p < 0.05) biofilm formation in vitro and on impregnated catheters. In addition, CPZ-impregnation potentiated the antibiofilm activity of ciprofloxacin and meropenem. Conclusion: These findings bring perspectives for the use of CPZ as an adjuvant for preventing and treating catheter-associated urinary tract infections.

scholarly journals Poloxamer 338 Affects Cell Adhesion and Biofilm Formation in Escherichia coli: Potential Applications in the Management of Catheter-Associated Urinary Tract Infections

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 885
Author(s):  
Mariarita Stirpe ◽  
Benedetta Brugnoli ◽  
Gianfranco Donelli ◽  
Iolanda Francolini ◽  
Claudia Vuotto
Keyword(s):  
Escherichia Coli ◽  
Cell Adhesion ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Biofilm Formation ◽  
Attenuated Total Reflection ◽  
Water Contact ◽  
E Coli ◽  
Static Conditions ◽  
Tract Infections

Poloxamers are nontoxic, amphiphilic copolymers used in different formulations. Due to its surfactant properties, Poloxamer 338 (P388) is herein proposed as a strategy to avoid biofilm formation often causing recalcitrant catheter-associated urinary tract infections (CAUTI). The aim is to evaluate the ability of P388 coatings to affect the adhesion of Ec5FSL and Ec9FSL Escherichia coli strains on silicone urinary catheters. Attenuated total reflection infrared spectroscopy, atomic force microscopy, and static water contact angle measurement were employed to characterize the P388-coated silicone catheter in terms of amount of P388 layered, coating thickness, homogeneity, and hydrophilicity. In static conditions, the antifouling power of P388 was defined by comparing the E. coli cells adherent on a hydrophilic P388-adsorbed catheter segment with those on an uncoated one. A P388-coated catheter, having a homogeneous coverage of 35 nm in thickness, reduced of 0.83 log10 and 0.51 log10 the biofilm of Ec5FSL and Ec9FSL, respectively. In dynamic conditions, the percentage of cell adhesion on P388-adsorbed silicone channels was investigated by a microfluidic system, simulating the in vivo conditions of catheterized patients. As a result, both E. coli isolates were undetected. The strong and stable antifouling property against E. coli biofilm lead us to consider P388 as a promising anti-biofilm agent for CAUTIs control.

scholarly journals Escherichia coli CFT073 Fitness Factors during Urinary Tract Infection: Identification Using an Ordered Transposon Library

2020 ◽  
Vol 86 (13) ◽  
Author(s):  
Allyson E. Shea ◽  
Juan Marzoa ◽  
Stephanie D. Himpsl ◽  
Sara N. Smith ◽  
Lili Zhao ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Urinary Tract Infections ◽  
Human Urine ◽  
Transposon Mutagenesis ◽  
Content Type ◽  
E Coli ◽  
Tract Infections

ABSTRACT Urinary tract infections (UTI), the second most diagnosed infectious disease worldwide, are caused primarily by uropathogenic Escherichia coli (UPEC), placing a significant financial burden on the health care system. High-throughput transposon mutagenesis combined with genome-targeted sequencing is a powerful technique to interrogate genomes for fitness genes. Genome-wide analysis of E. coli requires random libraries of at least 50,000 mutants to achieve 99.99% saturation; however, the traditional murine model of ascending UTI does not permit testing of large mutant pools due to a bottleneck during infection. To address this, an E. coli CFT073 transposon mutant ordered library of 9,216 mutants was created and insertion sites were identified. A single transposon mutant was selected for each gene to assemble a condensed library consisting of 2,913 unique nonessential mutants. Using a modified UTI model in BALB/c mice, we identified 36 genes important for colonizing the bladder, including purB, yihE, and carB. Screening of the condensed library in vitro identified yigP and ubiG to be essential for growth in human urine. Additionally, we developed a novel quantitative PCR (qPCR) technique to identify genes with fitness defects within defined subgroups of related genes (e.g., genes encoding fimbriae, toxins, etc.) following UTI. The number of mutants within these subgroups circumvents bottleneck restriction and facilitates validation of multiple mutants to generate individual competitive indices. Collectively, this study investigates the bottleneck effects during UTI, provides two techniques for evading those effects that can be applied to other disease models, and contributes a genetic tool in prototype strain CFT073 to the field. IMPORTANCE Uropathogenic Escherichia coli strains cause most uncomplicated urinary tract infections (UTI), one of the most common infectious diseases worldwide. Random transposon mutagenesis techniques have been utilized to identify essential bacterial genes during infection; however, this has been met with limitations when applied to the murine UTI model. Conventional high-throughput transposon mutagenesis screens are not feasible because of inoculum size restrictions due to a bottleneck during infection. Our study utilizes a condensed ordered transposon library, limiting the number of mutants while maintaining the largest possible genome coverage. Screening of this library in vivo, and in human urine in vitro, identified numerous candidate fitness factors. Additionally, we have developed a novel technique using qPCR to quantify bacterial outputs following infection with small subgroups of transposon mutants. Molecular approaches developed in this study will serve as useful tools to probe in vivo models that are restricted by anatomical, physiological, or genetic bottleneck limitations.

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