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.

scholarly journals Characterization of plasmid-mediated quinolone resistance by the qnrS gene in Escherichia coli isolated from healthy chickens and pigs

2009 ◽  
Vol 54 (No. 10) ◽  
pp. 473-482 ◽  
Author(s):  
H.-C. Kuo ◽  
C.-C. Chou ◽  
C. Tu ◽  
S.-R. Gong ◽  
C.-L. Han ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Blot Hybridization ◽  
Southern Blot Hybridization ◽  
Quinolone Resistance ◽  
E Coli ◽  
Sequence Variations ◽  
Southern Blot Hybridization Analysis ◽  
Ciprofloxacin Resistance ◽  
Polymerase Chain

The prevalence of <I>qnr</I> and <I>qepA</I> genes in 660 <I>Escherichia coli</I> isolates was investigated in healthy animals from 30 pig farms and 30 chicken farms in Taiwan from January 2005 to February 2006 by the polymerase chain reaction. The <I>qnrS</I> gene, but not <I>qnrA, qnrB, </I> and <I>qepA</I> were detected in 12/360 pig isolates (3.33%) and in 6/300 chicken isolates (2%). Southern blot hybridization analysis indicated that <I>qnrS</I> was located on plasmids ranging in size from 50–165 kb. Eleven of the 18 <I>qnrS</I> positive isolates which showed a high ciprofloxacin resistance phenotype (minimum inhibitory concentration ≥ 8 mg/l) also had amino acid sequence variations in chromosomal quinolone resistance-determining regions of <I>gyrA</I> and <I>parC</I>. Only two <I>qnrS</I>-positive isolates carried the <I>aac(6’)-Ib-cr</I>variant that mediates FQ acetylation. For the high percentage resistance of cephalosporins, the<I> bla</I><sub>CTX-M</sub> gene was also examined in <I>qnrS</I>-positive isolates. The <I>bla</I><sub>CTX-M</sub> gene was detected in fifteen isolates (15/18, 83.3%) of which 12 isolates were <I>bla</I><sub>CTX-M-1</sub> and three isolates were <I>bl</I><sub>CTX-M-15</sub>. This study demonstrated a close linkage between the <I>qnrS</I> gene and <I>bla</I><sub>CTX-M-1</sub>, suggesting CTX-M and Qnr-based mechanisms might be co-emerging in <I>E. coli</I> strains isolated from healthy chickens and pigs under selective pressure of quinolone and cephalosporine administration.

scholarly journals Resistance Profiling and Molecular Characterization of Extended-Spectrum/Plasmid-Mediated AmpC β-Lactamase-Producing Escherichia coli Isolated from Healthy Broiler Chickens in South Korea

2020 ◽  
Vol 8 (9) ◽  
pp. 1434 ◽  
Author(s):  
Hyun-Ju Song ◽  
Dong Chan Moon ◽  
Abraham Fikru Mechesso ◽  
Hee Young Kang ◽  
Mi Hyun Kim ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Phylogenetic Analysis ◽  
Resistance Gene ◽  
Broiler Chickens ◽  
Human Consumption ◽  
E Coli ◽  
Extended Spectrum ◽  
First Time ◽  
Pfge Profiles

We aimed to identify and characterize extended-spectrum β-lactamase (ESBL)-and/or plasmid-mediated AmpC β-lactamase (pAmpC)-producing Escherichia coli isolated from healthy broiler chickens slaughtered for human consumption in Korea. A total of 332 E. coli isolates were identified from 339 cloacal swabs in 2019. More than 90% of the isolates were resistant to multiple antimicrobials. ESBL/pAmpC-production was noted in 14% (46/332) of the isolates. Six of the CTX-M-β-lactamase-producing isolates were found to co-harbor at least one plasmid-mediated quinolone resistance gene. We observed the co-existence of blaCMY-2 and mcr-1 genes in the same isolate for the first time in Korea. Phylogenetic analysis demonstrated that the majority of blaCMY-2-carrying isolates belonged to subgroup D. Conjugation confirmed the transferability of blaCTX-M and blaCMY-2 genes, as well as non-β-lactam resistance traits from 60.9% (28/46) of the ESBL/pAmpC-producing isolates to a recipient E. coli J53. The ISECP, IS903, and orf477 elements were detected in the upstream or downstream regions. The blaCTX-M and blaCMY-2 genes mainly belonged to the IncI1, IncHI2, and/or IncFII plasmids. Additionally, the majority of ESBL/pAmpC-producing isolates exhibited heterogeneous PFGE profiles. This study showed that healthy chickens act as reservoirs of ESBL/pAmpC-producing E. coli that can potentially be transmitted to humans.

scholarly journals Characterization of a Novel Pyranopyridine Inhibitor of the AcrAB Efflux Pump of Escherichia coli

2013 ◽  
Vol 58 (2) ◽  
pp. 722-733 ◽  
Author(s):  
Timothy J. Opperman ◽  
Steven M. Kwasny ◽  
Hong-Suk Kim ◽  
Son T. Nguyen ◽  
Chad Houseweart ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Efflux Pump ◽  
Efflux Pumps ◽  
Gram Negative ◽  
Content Type ◽  
E Coli ◽  
Resistance Nodulation Division ◽  
Rnd Efflux Pumps ◽  
Acrab Efflux Pump

ABSTRACTMembers of the resistance-nodulation-division (RND) family of efflux pumps, such as AcrAB-TolC ofEscherichia coli, play major roles in multidrug resistance (MDR) in Gram-negative bacteria. A strategy for combating MDR is to develop efflux pump inhibitors (EPIs) for use in combination with an antibacterial agent. Here, we describe MBX2319, a novel pyranopyridine EPI with potent activity against RND efflux pumps of theEnterobacteriaceae. MBX2319 decreased the MICs of ciprofloxacin (CIP), levofloxacin, and piperacillin versusE. coliAB1157 by 2-, 4-, and 8-fold, respectively, but did not exhibit antibacterial activity alone and was not active against AcrAB-TolC-deficient strains. MBX2319 (3.13 μM) in combination with 0.016 μg/ml CIP (minimally bactericidal) decreased the viability (CFU/ml) ofE. coliAB1157 by 10,000-fold after 4 h of exposure, in comparison with 0.016 μg/ml CIP alone. In contrast, phenyl-arginine-β-naphthylamide (PAβN), a known EPI, did not increase the bactericidal activity of 0.016 μg/ml CIP at concentrations as high as 100 μM. MBX2319 increased intracellular accumulation of the fluorescent dye Hoechst 33342 in wild-type but not AcrAB-TolC-deficient strains and did not perturb the transmembrane proton gradient. MBX2319 was broadly active againstEnterobacteriaceaespecies andPseudomonas aeruginosa. MBX2319 is a potent EPI with possible utility as an adjunctive therapeutic agent for the treatment of infections caused by Gram-negative pathogens.

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 Characterization of proteobacterial plasmid integron-encoded qac efflux pump sequence diversity and quaternary ammonium compound antiseptic selection in E. coli grown planktonically and as biofilms

2021 ◽  
Author(s):  
Carmine J. Slipski ◽  
Taylor R. Jamieson-Datzkiw ◽  
George G. Zhanel ◽  
Denice C. Bay
Keyword(s):  
Escherichia Coli ◽  
Quaternary Ammonium ◽  
Susceptibility Testing ◽  
Efflux Pump ◽  
Multidrug Resistant ◽  
Quaternary Ammonium Compound ◽  
Ammonium Compound ◽  
E Coli ◽  
Susceptibility Patterns

Qac efflux pumps from proteobacterial multidrug-resistant plasmids are integron-encoded and confer resistance to quaternary ammonium compound (QAC) antiseptics, however, many are uncharacterized and misannotated. A survey of >2000 plasmid-encoded qac identified 37 unique qac sequences that correspond to one of five representative motifs: QacE, QacEΔ1, QacF/L, QacH/I, and QacG. Antimicrobial susceptibility testing of each cloned qac member in Escherichia coli , highlighted distinctive antiseptic susceptibility patterns that were most prominent when cells grew as biofilms.

scholarly journals Prevalence and Characterization of Plasmid-mediated Quinolone Resistance Genes among Escherichia coli Strains Isolated from Different Water Sources in Alborz Province, Iran

2019 ◽  
Vol 11 (1) ◽  
pp. 36-41
Author(s):  
Reza Ranjbar ◽  
Shahrzad Tavanania ◽  
Azar Sabokbar ◽  
Faham Khamesipour
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Surface Water ◽  
Nalidixic Acid ◽  
Bacterial Species ◽  
Water Sources ◽  
Quinolone Resistance ◽  
E Coli ◽  
Different Water Sources

BACKGROUND: This study was conducted to investigate the prevalence of quinolone resistance associated (qnr) antibiotic resistance among Escherichia coli strains isolated from different water sources in Alborz province, Iran.METHODS: E. coli strains were isolated and identified by standard microbiological and biochemical tests from surface water sources in Alborz province, Iran in 2013. Fluoroquinolone-resistant isolates were determined using the antimicrobial susceptibility test determined by the Kirby–Bauer assay. Total genomic and plasmid DNA were extracted by boiling method. The presence of qnr genes in all nalidixic-acid and ciprofloxacin-resistant E. coli strains was determined by Polymerase Chain Reaction (PCR). The PCR amplicons were visualized after electrophoresis stained with ethidium bromide.RESULTS: One hundred E. coli strains were isolated from the water sources examined in this study. As much as 22.7% and 7.3% of the isolates were resistant to nalidixic acid and ciprofloxacin respectively. While qnrS, qnrB and qnrA genes were detected in 28%, 9% and 1% of fluoroquinolone-resistant isolates respectively. All fluoroquinolone-susceptible isolates however did not contain any of the qnr genes.CONCLUSION: This study reflects an increasing prevalence of fluoroquinolone-resistant E. coli strains in surface water sources. Underlining the importance of surface water sources as reservoirs for dissemination of potentially pathogenic E. coli and horizontal gene transfer between other waterborne bacterial species. Other possible mechanisms of resistance should also be investigated for better characterization of quinolone-resistant E. coli isolates. Therefore, immediate measures are needed to control and treat water sources more effectively.KEYWORDS: antibiotic resistance, E. coli, qnr genes, water sources 

scholarly journals Risk factors for and mortality of extended-spectrum-β-lactamase-producing Klebsiella pneumoniae and Escherichia coli nosocomial bloodstream infections

2009 ◽  
Vol 51 (4) ◽  
pp. 211-216 ◽  
Author(s):  
Silvana Vargas Superti ◽  
Gustavo Augusti ◽  
Alexandre Prehn Zavascki
Keyword(s):  
Risk Factors ◽  
Escherichia Coli ◽  
Risk Factor ◽  
Klebsiella Pneumoniae ◽  
Bloodstream Infections ◽  
Blood Cultures ◽  
Esbl Production ◽  
E Coli ◽  
Extended Spectrum ◽  
Esbl Producers

A case-control study, involving patients with positive blood cultures for Klebsiella pneumoniae (KP) or Escherichia coli (EC) EC and controls with positive blood cultures for non-ESBL-KP or EC, was performed to assess risk factors for extended-spectrum-β-lactamase (ESBL) production from nosocomial bloodstream infections (BSIs). Mortality among patients with BSIs was also assessed. The study included 145 patients (81, 59.5% with K. pneumoniae and 64, 44.1% with E. coli BSI); 51 (35.2%) isolates were ESBL producers and 94 (64.8%) nonproducers. Forty-five (55.6%) K. pneumoniae isolates were ESBL producers, while only six (9.4%) E. coli isolates produced the enzyme. Multivariate analysis showed that recent exposure to piperacillin-tazobactam (adjusted Odds Ratio [aOR] 6.2; 95%CI 1.1-34.7) was a risk factor for ESBL BSI. K. pneumoniae was significantly more likely to be an ESBL-producing isolate than E. coli (aOR 6.7; 95%CI 2.3-20.2). No cephalosporin class was independently associated with ESBLs BSI; however, in a secondary model considering all oxymino-cephalosporins as a single variable, a significant association was demonstrated (aOR 3.7; 95%CI 1.3-10.8). Overall 60-day mortality was significantly higher among ESBL-producing organisms. The finding that piperacillin-tazobactam use is a risk factor for ESBL-production in KP or EC BSIs requires attention, since this drug can be recommended to limit the use of third-generation cephalosporins.

scholarly journals Characterization of sulphonamide-resistant Escherichia coli using comparison of sul2 gene sequences and multilocus sequence typing

Microbiology ◽  
2009 ◽  
Vol 155 (3) ◽  
pp. 831-836 ◽  
Author(s):  
Margarita Trobos ◽  
Henrik Christensen ◽  
Marianne Sunde ◽  
Steen Nordentoft ◽  
Yvonne Agersø ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Multilocus Sequence Typing ◽  
Point Mutations ◽  
Bloodstream Infections ◽  
Poultry Meat ◽  
E Coli ◽  
Pcr Products ◽  
Synonymous Change ◽  
Host Origin

The sul2 gene encodes sulphonamide resistance (SulR) and is commonly found in Escherichia coli from different hosts. We typed E. coli isolates by multilocus sequence typing (MLST) and compared the results to sequence variation of sul2, in order to investigate the relation to host origin of pathogenic and commensal E. coli strains and to investigate whether transfer of sul2 into different genomic lineages has happened multiple times. Sixty-eight E. coli isolated in Denmark and Norway from different hosts and years were MLST typed and sul2 PCR products were sequenced and compared. PFGE was performed in a subset of isolates. All isolates were divided into 45 different sequence types (STs), with clonal complexes CC10, CC23, CC168, CC350 and CC69 being the most frequent. The sul2 gene from the majority of E. coli strains had only two point mutations, at positions 159 and 197, leading to a synonymous and a non-synonymous change, respectively. Five strains had extra single mutations. All poultry, poultry meat, and Danish human blood isolates had the same sul2 ST and some of these strains clustered under the same MLST STs, indicating that they shared habitats. Most PFGE profiles clustered according to source, but some included different sources. SulR E. coli from different animals, food, human faeces and infections did not cluster according to their origin, suggesting that these habitats share E. coli and sul2 gene types. However, while pig isolates on one occasion clustered with urinary tract infection isolates, poultry isolates seemed more related to isolates from bloodstream infections in humans. Presence of mainly two types of the sul2 gene in both human and animal isolates, irrespective of date and geography, and the presence of both types in the same clonal lineages, suggest horizontal transfer of sul2.

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 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.

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