The influence of antibiotic resistance gene carriage on biofilm formation by two Escherichia coli strains associated with urinary tract infections

2014 ◽  
Vol 60 (2) ◽  
pp. 105-111 ◽  
Author(s):  
Amy Huei Teen Teh ◽  
Yi Wang ◽  
Gary A. Dykes
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Resistance Gene ◽  
Biofilm Formation ◽  
Antibiotic Resistance Gene ◽  
Kanamycin Resistance ◽  
Marker Genes ◽  
E Coli ◽  
Tract Infections

Urinary tract infections (UTI) caused by uropathogenic Escherichia coli are one of the most common forms of human disease. In this study, the effect of the presence of newly acquired antibiotic resistance genes on biofilm formation of UTI-associated E. coli strains was examined. Two clinical UTI-associated E. coli strains (SMC18 and SMC20) carrying different combinations of virulence genes were transformed with pGEM-T, pGEM-T::KmΔAmp, or pGEM-T::Km to construct ampicillin-resistant (KmSAmpR), kanamycin-resistant (KmRAmpS), or ampicillin- and kanamycin-resistant (KmRAmpR) strains. Transformed and wild-type strains were characterized for biofilm formation, bacterial surface hydrophobicity, auto-aggregation, morphology, and attachment to abiotic surfaces. Transformation with a plasmid carrying an ampicillin resistance gene alone decreased (p < 0.05) biofilm formation by SMC18 (8 virulence marker genes) but increased (p < 0.05) biofilm formation by SMC20 (5 virulence marker genes). On the other hand, transformation with a plasmid carrying a kanamycin resistance gene alone or both ampicillin and kanamycin resistance genes resulted in a decrease (p < 0.05) in biofilm formation by SMC18 but did not affect (p > 0.05) the biofilm formation by SMC20. Our results suggest that transformation of UTI-associated E. coli with plasmids carrying different antibiotic resistance gene(s) had a significant impact on biofilm formation and that these effects were both strain dependent and varied between different antibiotics.

scholarly journals Bacterial Profile of Urinary Tract Infections: Evaluation of Biofilm Formation and Antibiotic Resistance Pattern of Uropathogenic Escherichia coli

2020 ◽  
Vol 14 (4) ◽  
pp. 2577-2584
Author(s):  
Tariq Ahmad Shah ◽  
P. Preethishree ◽  
Ashwini ◽  
Vidya Pai
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Biofilm Formation ◽  
Urine Samples ◽  
Diffusion Method ◽  
E Coli ◽  
Biofilm Production ◽  
Tract Infections

Urinary tract infection (UTI) is one of the most common complaints in the outpatient clinic and a major health problem owing to the emergence of antibiotic resistance and biofilm formation. The objective of this study was to isolate and identify the causative bacterial agent of UTI and detect in vitro biofilm formation by Escherichia coli and investigate its correlation with antibiotic resistance. Urine samples from 519 patients with suspected UTIs were collected and processed by conventional microbiological procedures. Antimicrobial susceptibility testing for E. coli isolates was performed on Mueller Hinton agar (MHA) plates using the Kirby-Bauer disk diffusion method. Biofilm production was evaluated using the tissue culture plate method. Of 519 urine samples, 115 (22.1%) showed significant bacteriuria. The most common isolate was E. coli (n=57, 49.6%), followed by Klebsiella spp. (n=23, 20%). All E. coli isolates were evaluated for their ability to form biofilms in vitro. Of 57 isolates, 50 (87.7%) were biofilm producers and 7 (12.3%) were non-biofilm producers. Antibiogram of E. coli isolates revealed the highest resistance to ampicillin (96.5%) and nitrofurantoin (91.2%), followed by amoxyclav (82.5%), ceftazidime (73.7%), cefepime (71.9%), and tetracycline (71.9%). A significant association (p<0.05) was observed between biofilm formation and resistance to amoxyclav, ceftazidime, cefepime, imipenem, and nitrofurantoin. A significant correlation was noted between biofilm production and antibiotic resistance. Hence, screening of all isolates of uropathogenic E. coli for biofilm production and studying their antibiogram would allow appropriate choice of antibiotic therapy.

scholarly journals Evaluation of Immunocompetent Urinary Tract Infected Balb/C Mouse Model For the Study of Antibiotic Resistance Development Using Escherichia Coli CFT073 Infection

Antibiotics ◽  
2019 ◽  
Vol 8 (4) ◽  
pp. 170 ◽  
Author(s):  
Ashok Chockalingam ◽  
Sharron Stewart ◽  
Lin Xu ◽  
Adarsh Gandhi ◽  
Murali K. Matta ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Bladder Tissue ◽  
Resistance Development ◽  
Once Daily ◽  
E Coli ◽  
Low Levels ◽  
Tract Infections

Urinary tract infections (UTI) are common worldwide and are becoming increasingly difficult to treat because of the development of antibiotic resistance. Immunocompetent murine models of human UTI have been used to study pathogenesis and treatment but not for investigating resistance development after treatment with antibiotics. In this study, intravesical inoculation of uropathogenic Escherichia coli CFT073 in immunocompetent Balb/c mice was used as a model of human UTI. The value of the model in investigating antibiotic exposure on in vivo emergence of antibiotic resistance was examined. Experimentally infected mice were treated with 20 or 200 mg/kg ampicillin, 5 or 50 mg/kg ciprofloxacin, or 100 or 1000 mg/kg of fosfomycin. Ampicillin and ciprofloxacin were given twice daily at 8 h intervals, and fosfomycin was given once daily. Antibiotic treatment began 24 h after bacterial inoculation and ended after 72 h following the initial treatment. Although minimum inhibitory concentrations (MIC) for the experimental strain of E. coli were exceeded at peak concentrations in tissues and consistently in urine, low levels of bacteria persisted in tissues in all experiments. E. coli from bladder tissue, kidney, and urine grew on plates containing 1× MIC of antibiotic, but none grew at 3× MIC. This model is not suitable for studying emergent resistance but might serve to examine bacterial persistence.

scholarly journals Detection of biofilm among uropathogenic Escherichia coli and its correlation with antibiotic resistance pattern

2019 ◽  
Vol 11 (01) ◽  
pp. 017-022 ◽  
Author(s):  
Rashmi M. Karigoudar ◽  
Mahesh H. Karigoudar ◽  
Sanjay M. Wavare ◽  
Smita S. Mangalgi
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Biofilm Formation ◽  
Agar Plate ◽  
Diffusion Method ◽  
Resistance Pattern ◽  
Susceptibility Pattern ◽  
Sensitivity Testing ◽  
E Coli ◽  
Tract Infections

Abstract BACKGROUND: Escherichia coli accounts for 70%–95% of urinary tract infections (UTIs). UTI is a serious health problem with respect to antibiotic resistance and biofilms formation being the prime cause for the antibiotic resistance. Biofilm can restrict the diffusion of substances and binding of antimicrobials. In this context, the present study is aimed to perform in vitro detection of biofilm formation among E. coli strains isolated from urine and to correlate their susceptibility pattern with biofilm formation. MATERIALS AND METHODS: A total of 100 E. coli strains isolated from patients suffering from UTI were included in the study. The identification of E. coli was performed by colony morphology, Gram staining, and standard biochemical tests. The detection of biofilm was carried out by Congo Red Agar (CRA) method, tube method (TM), and tissue culture plate (TCP) method. Antimicrobial sensitivity testing was performed by Kirby–Bauer disc diffusion method on Muller–Hinton agar plate. RESULTS: Of the 100 E. coli strains, 49 (49%) and 51 (51%) were from catheterized and noncatheterized patients, respectively. Biofilm production was positive by CRA, TM, and TCP method were 49 (49%), 55 (55%), and 69 (69%), respectively. Biofilm producers showed maximum resistance to co-trimoxazole (73.9%), gentamicin (94.2%), and imipenem (11.6%) when compared to nonbiofilm producers. Significant association was seen between resistance to antibiotic and biofilm formation with a P = 0.01 (<0.05). CONCLUSION: A greater understanding of biofilm detection in E. coli will help in the development of newer and more effective treatment. The detection of biofilm formation and antibiotic susceptibility pattern helps in choosing the correct antibiotic therapy.

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 Expression of the Fluoroquinolones Efflux Pump Genes acrA and mdfA in Urinary Escherichia coli Isolates

2017 ◽  
Vol 66 (1) ◽  
pp. 25-30 ◽  
Author(s):  
Sarah M. Abdelhamid ◽  
Rania R. Abozahra
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Molecular Mechanisms ◽  
Efflux Pump ◽  
Efflux System ◽  
E Coli ◽  
Reverse Transcription Pcr ◽  
Tract Infections

Escherichia coli is one of the most frequent causes of urinary tract infections. Efflux system overexpression is reported to contribute to E. coli resistance to several antibiotics. Our aim in this study was to investigate the relation between antibiotic resistance and the expression of the efflux pump genes acrA and mdfA in E. coli by real-time reverse transcription-PCR. We tested the in vitro susceptibilities to 12 antibiotics in 28 clinical isolates of E. coli obtained from urine samples. We also determined the minimum inhibitory concentrations of levofloxacin to these samples. We then revealed significant correlations between the overexpression of both mdfA and acrA and MICs of levofloxacin. In conclusion, we demonstrated that the increased expression of efflux pump genes such as mdfA and acrA can lead to levofloxacin resistance in E. coli. These findings contribute to further understanding of the molecular mechanisms of efflux pump systems and how they contribute to antibiotic resistance.

scholarly journals Urine Microbial Landscape in Children with Urinary Tract Infections in Ryazan and Ryazan Region

Doctor Ru ◽  
2021 ◽  
Vol 20 (10) ◽  
pp. 48-53
Author(s):  
N.A. Belykh ◽  
◽  
S.V. Tereschenko ◽  
N.A. Anikeeva ◽  
S.S. Kantutis ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Klebsiella Pneumoniae ◽  
Urinary Tract Infections ◽  
Patient Specific ◽  
E Coli ◽  
Resistance Determinants ◽  
Study Results ◽  
Tract Infections

Study Objective: To study a spectrum of uropathogens and their sensitivity to antimicrobials in urinary tract infections (UTIs) in children in Ryazan and Ryazan Region. Study Design: retrospective study. Materials and Methods. We conducted a retrospective local laboratory monitoring of urinary microflora and analysed its sensitivity to antimicrobials in 111 patients aged 2 months to 17 years old who were undergoing traditional UI therapy in 2020. The study group comprised 75 (67.6%) girls and 36 (32.4%) boys. Pathogen isolation and type identification were performed using urine specimens collected in sterile disposable plastic containers prior to antimicrobial therapy. Material was delivered for analysis within 2 hours from collection. For testing of pathogen sensitivity to antimicrobials, we used the phenotyping diffusion test and an analytical test for carbapenems inactivation. Study Results. Prevailing causative agents of UIs were Escherichia coli (50.4%) and Klebsiella pneumoniae (14.4%). Resistance determinants were found in 9.0% and 2.7% of Е. соli and K. pneumoniae urological strains, respectively. The main mechanism of resistivity was production of wide spectrum plasmid β-lactamases. The highest activity in E. coli was demonstrated by generation III–IV cephalosporins, aminoglycosides, fosfomicin (100%), nitrofurantoin (91.3%), and aminopenicillins (76.1–86.9%). For K. pneumoniae, generation III–IV cephalosporins and aminoglycosides were most potent (100%). All resistant pathogens were sensitive to cefoperazone sulbactam, meropenem, imipenem, aminoglycosides (100%); tigecycline, nitrofurantoin, and fosfomicin were most potent against E. coli. Conclusion. Children with UIs in Ryazan Region had mostly gram-negative bacteria in their urine (85.6%), Enterobacteralеs (81.1%) being a prevailing type. Antimicrobials resistance determinants were quite rare (17.8%) in these urine isolates; all of them were class A ЕSBL producers. These characteristic features of antibiotic resistance of uropathogenic enterobacteria strains allow using β-lactam antibiotics in empiric initial treatment and emphasising the need in patient-specific selection of antimicrobials. Keywords: antibacterial therapy, antibiotic resistance, children, urinary tract infections, Escherichia coli, Klebsiella pneumoniae.

scholarly journals Detection of Phylogenetic Groups and Drug Resistance Genes of Escherichia coli Causing Urinary Tract Infection in Southwest Iran

2021 ◽  
Vol 14 (2) ◽  
Author(s):  
Mostafa Boroumand Boroumand ◽  
Mohsen Naghmachi ◽  
Mohammad Amin Ghatee
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Phylogenetic Group ◽  
Phylogenetic Groups ◽  
E Coli ◽  
Pcr Method ◽  
Tract Infections

Background: Many bacteria can cause urinary tract infections (UTIs), among which Escherichia coli is the most common causative agent. E. coli strains are divided into eight phylogenetic groups based on the new Quadroplex-PCR method, which are different in terms of patterns of resistance to antibiotics, virulence, and environmental characteristics. Objectives: This study aimed to determine the phylogenetic groups and the prevalence of drug resistance genes in E. coli strains causing UTIs. Methods: In this descriptive cross-sectional study, 129 E. coli isolates obtained from the culture of patients with UTIs were evaluated for phylogenetic groups using the new method of Clermont et al. The identification of phylogenetic groups and antibiotic resistance genes was performed using the multiplex polymerase chain reaction (PCR) method. Results: In this study, concerning the distribution of phylogenetic groups among E. coli isolates, the phylogenetic group B2 (36.4%) was the most common phylogenetic group, followed by phylogroups C (13.2%), clade I (10.1%), D (9.3%), and A (3.1%) while groups B1 and F were not observed in any of the isolates, and 20.2% had an unknown state. Also, out of 129 E. coli isolates, the total frequency of tetA, tetB, sul1, sul2, CITM, DfrA, and qnr resistance genes was 59.7%, 66.7, 69, 62, 30.2, 23.3, and 20.2%, respectively. In this study, there was a significant relationship between antibiotics (P = 0.026), cefotaxime (P = 0.003), and nalidixic acid (P = 0.044) and E. coli phylogenetic groups. No significant relationship was observed between E. coli phylogenetic groups and antibiotic resistance genes. Conclusions: The results of this study showed that strains belonging to group B2 had the highest prevalence among other phylogroups, and also, the frequency of antibiotic resistance genes and drug-resistant isolates had a higher prevalence in this phylogroup. These results show that phylogroup B2 has a more effective role in causing urinary tract infections compared to other phylogroups, and this phylogroup can be considered a genetic reservoir of antibiotic resistance.

scholarly journals Genomic surveillance of Escherichia coli ST131 identifies local expansion and serial replacement of subclones

2019 ◽  
Author(s):  
Catherine Ludden ◽  
Arun Gonzales Decano ◽  
Dorota Jamrozy ◽  
Derek Pickard ◽  
Dearbhaile Morris ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Gene ◽  
Antibiotic Resistance Gene ◽  
Gene Variants ◽  
Term Care ◽  
E Coli ◽  
Esbl Gene ◽  
Combining Data ◽  
Clade C

AbstractEscherichia coli sequence type 131 (ST131) is a pandemic clone that is evolving rapidly with increasing levels of antimicrobial resistance. Here, we investigated an outbreak of E. coli ST131 producing extended spectrum β-lactamases (ESBLs) in a long-term care facility (LTCF) in Ireland by combining data from this LTCF (n=69) with other Irish (n=35) and global (n=690) ST131 genomes to reconstruct the evolutionary history and understand changes in population structure and genome architecture over time. This required a combination of short and long-read genome sequencing, de novo assembly, read mapping, ESBL gene screening, plasmid alignment and temporal phylogenetics. We found that clade C was the most prevalent (686 out of 794 isolates, 86%) of the three major ST131 clades circulating worldwide (A, B, C), and was associated with the presence of different ESBL alleles, diverse plasmids and transposable elements. Clade C was estimated to have emerged in ∼1985 and subsequently acquired different ESBL gene variants (blaCTX-M-14 vs blaCTX-M-15). An ISEcp1- mediated transposition of the blaCTX-M-15 gene further increased the diversity within Clade C. We discovered a local clonal expansion of a rare C2 lineage (C2_8) with a chromosomal insertion of blaCTX-M-15 at the mppA gene. This was acquired from an IncFIA plasmid. The C2_8 lineage clonally expanded in the Irish LTCF from 2006, displacing the existing C1 strain (C1_10) and highlighting the potential for novel ESBL-producing ST131 with a distinct genetic profile to cause outbreaks strongly associated with specific healthcare environments.ImportanceExtraintestinal pathogenic E. coli (ExPEC) ST131 is adapting in the context of antibiotic exposure, resulting in a pandemic with distinct genetic subtypes. Here, we track the evolution of antibiotic-resistance gene variants originally discovered in an ExPEC ST131 outbreak that was identified in a LTCF in Ireland. Analysis of 794 global ST131 genomes show that subclade C1 was associated with the initial infection outbreak, but that a new lineage from subclade C2 successfully displaced C1. This genetically distinct C2 subclade with a chromosomal insertion of a key antibiotic-resistance gene had clonally expanded within the LTCF. We provide new insights into the timing of genetic events driving the diversification of C2 subclades to show that that outbreak C2 strain likely evolved elsewhere before spreading to the LTCF. This study highlights the scope of antibiotic-resistance gene rearrangement within ST131, reinforcing the need to integrate genomic, epidemiological and microbiological approaches to understand ST131 transmission.

scholarly journals Selection and co-selection of antibiotic resistances among Escherichia coli by antibiotic use in primary care: an ecological analysis

2019 ◽  
Author(s):  
Koen B Pouwels ◽  
Berit Muller-Pebody ◽  
Timo Smieszek ◽  
Susan Hopkins ◽  
Julie V Robotham
Keyword(s):  
Escherichia Coli ◽  
Primary Care ◽  
Antibiotic Resistance ◽  
Urinary Tract ◽  
Respiratory Tract ◽  
Urinary Tract Infections ◽  
Respiratory Tract Infections ◽  
Antibiotic Use ◽  
E Coli ◽  
Tract Infections

AbstractThe majority of studies that link antibiotic usage and resistance focus on simple associations between the resistance against a specific antibiotic and the use of that specific antibiotic. However, the relationship between antibiotic use and resistance is more complex. Here we evaluate which antibiotics, including those mainly prescribed for respiratory tract infections, are associated with increased resistance among Escherichia coli isolated from urinary samples.Monthly primary care prescribing data were obtained from National Health Service (NHS) Digital. Positive E. coli records from urine samples in English primary care (n=888,207) between April 2014 and January 2016 were obtained from the Second Generation Surveillance System. Elastic net regularization was used to evaluate associations between prescribing of different antibiotic groups and resistance against amoxicillin, cephalexin, ciprofloxacin, co-amoxiclav and nitrofurantoin at the clinical commissioning group (CCG) level. England is divided into 209 CCGs, with each NHS practice prolonging to one CCG.Amoxicillin prescribing (measured in DDD/ 1000 inhabitants / day) was positively associated with amoxicillin (RR 1.03, 95% CI 1.01 – 1.04) and ciprofloxacin (RR 1.09, 95% CI 1.04 – 1.17) resistance. In contrast, nitrofurantoin prescribing was associated with lower levels of resistance to amoxicillin (RR 0.92, 95% CI 0.84 – 0.97). CCGs with higher levels of trimethoprim prescribing also had higher levels of ciprofloxacin resistance (RR 1.34, 95% CI 1.10 – 1.59).Amoxicillin, which is mainly (and often unnecessarily) prescribed for respiratory tract infections is associated with increased resistance against various antibiotics among E. coli causing urinary tract infections. Our findings suggest that when predicting the potential impact of interventions on antibiotic resistances it is important to account for use of other antibiotics, including those typically used for other indications.Author summaryAntibiotic resistance is increasingly recognised as a threat to modern healthcare. Effective antibiotics are crucial for treatment of serious bacterial infections and are necessary to avoid that complicated surgical procedures and chemotherapy becoming life-threatening. Antibiotic use is one of the main drivers of antibiotic resistance. The majority of antibiotic prescriptions are prescribed in primary care, however, a large proportion of these antibiotic prescriptions are unnecessary. Understanding which antibiotics are causing antibiotic resistance to what extent is needed to prevent under- or over-investment in interventions lowering use of specific antibiotics, such as rapid diagnostic tests for respiratory tract infection.We have statistically evaluated which antibiotics are associated with higher and lower levels of antibiotic resistance against common antibiotics among Escherichia coli bacteria sampled from the urinary tract by comparing antibiotic prescribing and resistance in different geographical areas in England. Our model shows that amoxicillin, the most commonly used antibiotic in England and mainly used for respiratory tract infections, is associated with increased resistance against several other antibiotics among bacteria causing urinary tract infections. The methods used in this study, that overcome several of the limitations of previous studies, can be used to explore the complex relationships between antibiotic use and antibiotic resistance in other settings.

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