scholarly journals Characterization of Integrons and Quinolone Resistance in Clinical Escherichia coli Isolates in Mansoura City, Egypt

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Shaymaa H. Abdel-Rhman ◽  
Rehab M. Elbargisy ◽  
Dina E. Rizk
Keyword(s):  
Escherichia Coli ◽  
Resistance Genes ◽  
Efflux Pump ◽  
Quinolone Resistance ◽  
Aminoglycoside Resistance ◽  
Variable Regions ◽  
Active Efflux ◽  
E Coli ◽  
High Prevalence ◽  
Class I Integrons

Escherichia coli is a common pathogen in both humans and animals. Quinolones are used to treat infections caused by Gram-negative bacteria, but resistance genes emerged. Only scarce studies investigated the association between plasmid-mediated quinolone resistance (PMQR) genes and integrons in clinical isolates of E. coli. The current study investigated the prevalence of quinolone resistance and integrons among 134 clinical E. coli isolates. Eighty (59.70%) isolates were quinolone-resistant, and 60/134 (44.77%) isolates were integron positive with the predominance of class I integrons (98.33%). There was a significant association between quinolone resistance and the presence of integrons ( P < 0.0001 ). Isolates from Urology and Nephrology Center and Gastroenterology Hospital were significantly quinolone-resistant and integron positive ( P ≤ 0.0005 ). Detection of PMQR genes on plasmids of integron-positive isolates showed that the active efflux pump genes oqxAB and qepA had the highest prevalence (72.22%), followed by the aminoglycoside acetyltransferase gene (aac(6′)-Ib-cr, 66.67%) and the quinolone resistance genes (qnr, 61.11%). Amplification and sequencing of integrons’ variable regions illustrated that no quinolone resistance genes were detected, and the most predominant gene cassettes were for trimethoprim and aminoglycoside resistance including dfrA17, dfrB4, and dfrA17-aadA5. In conclusion, this study reported the high prevalence of PMQR genes and integrons among clinical E. coli isolates. Although PMQR genes are not cassette-born, they were associated with integrons’ presence, which contributes to the widespread of quinolone resistance in Egypt.

scholarly journals New topoisomerase inhibitors: Evaluating the potency of gepotidacin and zoliflodacin in fluoroquinolone-resistant Escherichia coli upon tolC inactivation and differentiating their efflux pump substrate nature.

2020 ◽  
Author(s):  
Sabine Schuster ◽  
Martina Vavra ◽  
Raphael Köser ◽  
John W. A. Rossen ◽  
Winfried V. Kern
Keyword(s):  
Escherichia Coli ◽  
Efflux Pump ◽  
Multidrug Resistant ◽  
Quinolone Resistance ◽  
Topoisomerase Inhibitors ◽  
E Coli ◽  
Sequence Types ◽  
Substrate Nature

Inactivating tolC in multidrug-resistant Escherichia coli with differing sequence types and quinolone resistance-determining mutations reveals remarkably potentiated activity of the first-in-class topoisomerase inhibitors gepotidacin and zoliflodacin. Differences between both structurally unrelated compounds in comparison to fluoroquinolones regarding the selectivity of E. coli RND-type transporters, efflux inhibitors, and AcrB porter domain mutations were demonstrated. The findings should reinforce efforts to develop efflux bypassing drugs and provide AcrB targets with critical relevance for this purpose.

scholarly journals Virulence and plasmidic resistance determinants of Escherichia coli isolated from municipal and hospital wastewater treatment plants

2014 ◽  
Vol 13 (2) ◽  
pp. 311-318 ◽  
Author(s):  
Vera Calhau ◽  
Catarina Mendes ◽  
Angelina Pena ◽  
Nuno Mendonça ◽  
Gabriela Jorge Da Silva
Keyword(s):  
Escherichia Coli ◽  
Wastewater Treatment ◽  
Wastewater Treatment Plants ◽  
Multidrug Resistant ◽  
Quinolone Resistance ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Hospital Wastewater ◽  
Aminoglycoside Resistance ◽  
E Coli

Escherichia coli is simultaneously an indicator of water contamination and a human pathogen. This study aimed to characterize the virulence and resistance of E. coli from municipal and hospital wastewater treatment plants (WWTPs) in central Portugal. From a total of 193 isolates showing reduced susceptibility to cefotaxime and/or nalidixic acid, 20 E. coli with genetically distinct fingerprint profiles were selected and characterized. Resistance to antimicrobials was determined using the disc diffusion method. Extended spectrum β-lactamase and plasmid-mediated quinolone resistance genes, phylogroups, pathogenicity islands (PAIs) and virulence genes were screened by polymerase chain reaction (PCR). CTX-M producers were typed by multilocus sequence typing. Resistance to beta-lactams was associated with the presence of blaTEM,blaSHV, blaCTX-M-15 and blaCTX-M-32. Plasmid-mediated quinolone resistance was associated with qnrA, qnrS and aac(6′)-Ib-cr. Aminoglycoside resistance and multidrug-resistant phenotypes were also detected. PAI IV536, PAI IICFT073, PAI II536 and PAI ICFT073, and uropathogenic genes iutA, papAH and sfa/foc were detected. With regard to the clinical ST131 clone, it carried blaCTX-M-15, blaTEM-type, qnrS and aac(6′)-lb-cr; IncF and IncP plasmids, and virulence factors PAI IV536, PAI ICFT073, PAI IICFT073, iutA, sfa/foc and papAH were identified in the effluent of a hospital plant. WWTPs contribute to the dissemination of virulent and resistant bacteria in water ecosystems, constituting an environmental and public health risk.

scholarly journals Prevalence and Mechanism of Fluoroquinolone Resistance in Escherichia coli Isolated from Swine Feces in Korea

2017 ◽  
Vol 80 (7) ◽  
pp. 1145-1151 ◽  
Author(s):  
Yoon Sung Hu ◽  
Sook Shin ◽  
Yong Ho Park ◽  
Kun Taek Park
Keyword(s):  
Escherichia Coli ◽  
Resistance Genes ◽  
Resistance Mechanisms ◽  
Quinolone Resistance ◽  
Feed Additive ◽  
Diffusion Method ◽  
Resistant Bacteria ◽  
Double Mutation ◽  
Fecal Samples ◽  
E Coli

ABSTRACT In this study, we investigated the prevalence and fluoroquinolone (FQ) resistance mechanisms in Escherichia coli isolated from swine fecal samples. E. coli isolates were collected from 171 (72.2%) of 237 swine fecal samples. Of these, 59 isolates (34.5%) were confirmed as FQ-resistant E. coli by the disk diffusion method. Of the FQ-resistant isolates, three major FQ resistance mechanisms were investigated. Of the 59 isolates, plasmid-mediated quinolone resistance genes were detected in 9 isolates (15.3%). Efflux pump activity was found in 56 isolates (94.9%); however, this was not correlated with the increased FQ resistance measured by determining the MIC. Point mutations in quinolone resistance–determining regions were the main cause of FQ resistance. All 59 ciprofloxacin-resistant isolates had mutations in quinolone resistance–determining regions; of these 59 isolates, all (100%) had mutations in gyrA, 58 (98.3%) had mutations in parC, 22 (37.3%) had mutations in parE, and none had mutations in gyrB. The predominant mutation type was double mutation in gyrA (Ser83Leu plus mutation in aspartic acid 87), and all FQ-resistant isolates (except one) that had mutations in parC or parE also had double mutations in gyrA. Importantly, the frequencies of multidrug-resistant and extended-spectrum β-lactamase–producing E. coli were significantly higher in the high ciprofloxacin MIC group in this study. Compared with previous studies in Korea, the prevalence of FQ resistance and plasmid-mediated quinolone resistance genes had increased considerably in swine. Although the use of FQ as a feed additive is prohibited in Korea, use for self-treatment and therapeutic purposes has been increasing, which may be responsible for the higher FQ resistance rate observed in this study. Therefore, prudent use of FQ on animal farms is warranted to reduce the evolution of FQ-resistant bacteria in the animal industry.

Trend of plasmid-mediated quinolone resistance genes at the Children’s Hospital in Tunisia

2014 ◽  
Vol 63 (2) ◽  
pp. 195-202 ◽  
Author(s):  
Nour El-Houda Jlili ◽  
Samia Réjiba ◽  
Hanen Smaoui ◽  
Thomas Guillard ◽  
Françoise Chau ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Resistance Genes ◽  
Random Amplified Polymorphic Dna ◽  
Enterobacter Cloacae ◽  
Quinolone Resistance ◽  
Children's Hospital ◽  
Extended Spectrum ◽  
High Prevalence ◽  
Children’S Hospital ◽  
Dna Pcr

The prevalence of plasmid-mediated quinolone resistance genes [qnr, aac(6′)-Ib-cr and qepA] was sought among Enterobacteriaceae strains obtained from the Children’s Hospital of Tunis (Tunisia). Non-duplicate isolates (n = 278) with resistance to extended-spectrum cephalosporins and collected in 2003, 2007, 2008 and 2009 were screened for qnr genes. Forty (14.4 %) isolates were qnr positive and were screened for the presence of the aac(6′)-Ib-cr and qepA genes. qnrB was detected in 21 Klebsiella pneumoniae, 11 Escherichia coli and 6 Enterobacter cloacae isolates. Sequence analysis of the qnrB amplicons revealed variants including 24 qnrB1, 11 qnrB2 and 3 qnrB6. qnrS (qnrS1 allele) was detected only in K. pneumoniae isolates, either alone (two isolates) or with the qnrB gene (one isolate). The qnrA, qnrC and qnrD genes were not found in any of the 278 isolates. No qnr-positive isolates carried the qepA gene. Pyrosequencing results showed that aac(6′)-Ib-cr, a variant of the aac(6′)-Ib gene, was present in 31 qnr-positive isolates (21 K. pneumoniae isolates, seven Escherichia coli isolates and three Enterobacter cloacae isolates). aac(6′)-Ib was also found either alone (two isolates) or in association with aac(6′)-Ib-cr (one isolate). Of the 40 qnr-positive isolates, 92.5, 82.5, 57.5, 85 and 82.5 % were non-susceptible to nalidixic acid, ciprofloxacin, levofloxacin, ofloxacin and norfloxacin, respectively, and all were extended-spectrum β-lactamase producers. Random amplified polymorphic DNA-PCR typing of these isolates showed 16, 8 and 5 different genotypes in K. pneumoniae, Escherichia coli and Enterobacter cloacae isolates, respectively. Our study highlights the high prevalence of qnr in association with aac(6′)-Ib-cr among Enterobacteriaceae isolates, even from children, who are patients not overtreated with quinolones.

scholarly journals Characterization of uropathogenic ESBL-producing Escherichia coli isolated from hospitalized patients in western Algeria

2019 ◽  
Vol 13 (04) ◽  
pp. 291-302 ◽  
Author(s):  
Fatima Zenati ◽  
Abouddihaj Barguigua ◽  
Kaotar Nayme ◽  
Fethi Benbelaïd ◽  
Abdelmounaïm Khadir ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Characterization ◽  
Resistance Genes ◽  
Hospitalized Patients ◽  
Quinolone Resistance ◽  
High Rate ◽  
Phylogenetic Groups ◽  
E Coli ◽  
Tract Infections

Introduction: The aim of this study is to assess the prevalence and molecular characterization of uropathogenic Extended spectrum β-lactamases (ESBLs) producing Escherichia coli. Methodology: During 3 years, all hospitalized patients at the University-affiliated hospital of Tlemcen and presenting urinary tract infections caused by E. coli were considered as potential study participants. These E. coli isolates were examined phenotypically for ESBL production. ESBL strains were subjected to antimicrobial susceptibility testing and were investigated for the presence of plasmid mediated quinolone resistance genes, 16SrRNA methylase genes and virulence genes by using conventional PCR and DNA sequencing. The molecular characterization of ESBL strains was established by phylogenetic grouping method and ERIC-PCR. Results: The overall prevalence of ESBL was 32.5%. The blaCTX-M-15 was the most frequently detected in ESBL isolates, followed by blaCTX-M-14, blaCTX-M-28, blaCTX-M-1 and blaSHV-12 respectively. The plasmid-mediated quinolone resistance genes were detected in the 15 ESBL strains with the aac(6’)-Ib-cr gene was the most detected followed by qnrB1 and qnrA1 gene respectively. Among the 22 ESBL isolates resistant to gentamicin and amikacin, the 16SrRNA methylase genes were detected in 4 isolates. The sfa and pap virulent genes were founds in 26% and 22% of isolates receptively. The genotyping analysis of all strains revealed that almost were belonged to phylogenetic groups A1 and A0 and fourteen distinct clones. Conclusion: The emergence of uropathogenic ESBL isolates and the high rate of blaCTX-M are alarming in Algeria. Strict measure must be required to control the further spread of these strains in Algerian hospitals.

scholarly journals Molecular Characterization of gyrA, parC and qepA Genes in Quinolone Resistant ESBL Producing E. Coli Isolated from Patients in HTAA, Kuantan

2020 ◽  
Vol 18 (2) ◽  
Author(s):  
Rahmatullah Sirat ◽  
Hairul Aini Hamzah ◽  
Mohammed Imad A. Mustafa Mahmud ◽  
Siti Nurliyana Binti Ahmad
Keyword(s):  
Resistance Genes ◽  
Antibiotic Susceptibility ◽  
Efflux Pump ◽  
Molecular Techniques ◽  
Quinolone Resistance ◽  
Strong Positive Correlation ◽  
Resistance Mutations ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Antibiotic Susceptibility Test

Introduction: Quinolone resistance and extended spectrum beta lactamase production has increased in  E.coli  and considered a serious problem worldwide. It is worth to monitor resistance mechanism in  E.coli  to provide guidance for optimizing antimicrobial treatments,control and spread of resistance. The objective of this study was to molecularly characterize  gyrA, parC  genes and  plasmid mediated  qepA  efflux pump gene, in QR-ESBL  E. coli  isolates obtained from patients in HTAA, Kuantan. The antibiotic susceptibility profile was also studied. Materials and Method: 32 QR-ESBL and six quinolone-susceptible  E. coli  isolates from  September 30 November,2018) included in the study. The isolates were reconfirmed with known phenotypic tests and antibiotic susceptibility test was performed. PCR and DNA sequencing were performed for the identification of mutations in quinolone resistance determining region. Result: Resistance to ampicillin, tetracycline, nalidixic acid was (100%) followed by cefotaxime (96.9%), ciprofloxacin (78.1%) trimethoprim sulfamethoxazole (75%), ceftazidime (56.3%), cefepime (43.8%) and gentamycin (25%). None of the isolates was resistant to piperacillin-tazobactam, amikacin, imipenem, meropenem, ertapenem, and colistin. PCR successfully amplified the  gyrA  and  parC  genes, however,  qepA  gene was not detected by PCR in the isolates. Majority of the isolates had point mutation in (QRDR) of GyrA at codons 83 and 87 and in ParC at codons 80 and 84. Two isolates had mutations outside of QRDR at codons 144 and 167 in ParC. Strong positive correlation was found between MIC levels of ciprofloxacin and the number of resistance mutations. Sequencing of 6 (QS-ESBL)  E. coli  revealed absence of resistance mutations.  Conclusion: Quinolone resistance in the isolates was mainly due to mutations in  gyrA, ParC  genes. Acquisition of multidrug resistance genes through innate gene mutations and mobile genetic elements contribute to the emergence of (MDR). This study reinforces the importance of being vigilant in utilizing molecular techniques to monitor for emergence of resistance genes in different locations.

scholarly journals Emergence of Plasmid-Mediated Quinolone Resistance in Escherichia coli in Europe

2005 ◽  
Vol 49 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Hedi Mammeri ◽  
Marc Van De Loo ◽  
Laurent Poirel ◽  
Luis Martinez-Martinez ◽  
Patrice Nordmann
Keyword(s):  
Escherichia Coli ◽  
Topoisomerase Ii ◽  
Antibiotic Resistance Genes ◽  
Quinolone Resistance ◽  
Conjugative Plasmid ◽  
Aminoglycoside Resistance ◽  
Class 1 Integron ◽  
Promoter Sequences ◽  
E Coli ◽  
Class 1

ABSTRACT Although quinolone resistance results mostly from chromosomal mutations, it may also be mediated by a plasmid-encoded qnr gene in members of the family Enterobacteriaceae. Thus, 297 nalidixic-acid resistant strains of 2,700 Escherichia coli strains that had been isolated at the Bicêtre Hospital (Le Kremlin-Bicêtre, France) in 2003 were screened for qnr by PCR. A single E. coli isolate that carried a ca. 180-kb conjugative plasmid encoding a qnr determinant was identified. It conferred low-level resistance to quinolones and was associated with a chromosomal mutation in subunit A of the topoisomerase II gene. The qnr gene was located on a sul1-type class 1 integron just downstream of a conserved region (CR) element (CR1) comprising the Orf513 recombinase. Promoter sequences for qnr expression overlapped the extremity of CR1, indicating the role of CR1 in the expression of antibiotic resistance genes. This integron was different from other qnr-positive sul1-type integrons identified in American and Chinese enterobacterial isolates. In addition, plasmid pQR1 carried another class 1 integron that was identical to In53 from E. coli. The latter integron possessed a series of gene cassettes, including those coding for the extended-spectrum β-lactamase VEB-1, the rifampin ADP ribosyltransferase ARR-2, and several aminoglycoside resistance markers. This is the first report of plasmid-mediated quinolone resistance in Europe associated with an unknown level of plasmid-mediated multidrug resistance in Enterobacteriaceae.

scholarly journals Involvement of Antibiotic Efflux Machinery in Glutathione-Mediated Decreased Ciprofloxacin Activity in Escherichia coli

2016 ◽  
Vol 60 (7) ◽  
pp. 4369-4374 ◽  
Author(s):  
Manish Goswami ◽  
Mahesh Subramanian ◽  
Ranjeet Kumar ◽  
Jana Jass ◽  
Narendra Jawali
Keyword(s):  
Escherichia Coli ◽  
Efflux Pump ◽  
Antibacterial Action ◽  
Potential Mechanism ◽  
Content Type ◽  
Active Efflux ◽  
E Coli ◽  
Acrab Efflux Pump ◽  
Antibiotic Efflux

ABSTRACTWe have analyzed the contribution of different efflux components to glutathione-mediated abrogation of ciprofloxacin's activity inEscherichia coliand the underlying potential mechanism(s) behind this phenomenon. The results indicated that glutathione increased the total active efflux, thereby partially contributing to glutathione-mediated neutralization of ciprofloxacin's antibacterial action inE. coli. However, the role of glutathione-mediated increased efflux becomes evident in the absence of a functional TolC-AcrAB efflux pump.

Phenotypic and genotypic quinolone resistance in Escherichia coli underlining GyrA83/87 mutations as a target to detect ciprofloxacin resistance

2020 ◽  
Vol 75 (9) ◽  
pp. 2466-2470
Author(s):  
Anaëlle Muggeo ◽  
Emmanuelle Cambau ◽  
Marlène Amara ◽  
Maïté Micaëlo ◽  
Béatrice Pangon ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Efflux Pump ◽  
Bloodstream Infections ◽  
Blood Cultures ◽  
Quinolone Resistance ◽  
E Coli ◽  
Molecular Determinants ◽  
Ciprofloxacin Resistance ◽  
First Time

Abstract Background Quinolone resistance (QR) is one component of the MDR emerging in Escherichia coli and is of particular concern given the widespread use of fluoroquinolones. Objectives To characterize the QR phenotypes and genotypes in E. coli responsible for bloodstream infections and to propose molecular determinants that could be targeted to predict ciprofloxacin resistance. Methods E. coli isolates from blood cultures in three French hospitals were studied for quinolone MICs and characterization of genotypic QR determinants (QRg). Results Among 507 isolates tested for MICs, 148 (29.2%) were resistant to quinolones based on EUCAST breakpoints and 143 (28.2%) harboured at least one QRg. QRg were mainly mutations in the QRDR (138 isolates, 27.2%), with 55.8% of these isolates carrying at least three QRDR mutations. gyrA mutations predominated (92.8%) followed by parC (61.6%), parE (32.6%) and gyrB (1.4%) mutations. Only 4.7% of the isolates harboured a plasmid-mediated quinolone resistance (PMQR) gene: aac(6′)-Ib-cr (60.0%) or qnr (qnrS, qnrB) (32.0%). For the first time in France, we reported the qepA4 allele of the plasmid-encoded efflux pump QepA. Only five isolates carried PMQR without a QRDR mutation. The positive predictive value (PPV) for ciprofloxacin resistance was 100% for any QRg and 99.2% for gyrA mutations specifically. Conclusions QR observed in E. coli isolates involved in bloodstream infections is still mainly due to QRDR mutations, especially at codons GyrA83/87, which could be used as a molecular target to rapidly detect resistance.

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