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 Molecular Characterization of Plasmid-Mediated Non-O157 Verotoxigenic Escherichia coli Isolated from Infants and Children with Diarrhea

2020 ◽  
Vol 17 (3) ◽  
pp. 0710
Author(s):  
Md Fazlul Karim Khan ◽  
Shah Samiur Rashid
Keyword(s):  
Escherichia Coli ◽  
Virulence Gene ◽  
Multidrug Resistant ◽  
Plasmid Curing ◽  
Health Issues ◽  
E Coli ◽  
Disk Diffusion Assay ◽  
Microbiological Techniques ◽  
Polymerase Chain

A significant increase in the incidence of non-O157 verotoxigenic Escherichia coli (VTEC) infections have become a serious health issues, and this situation is worsening due to the dissemination of plasmid mediated multidrug-resistant microorganisms worldwide. This study aims to investigate the presence of plasmid-mediated verotoxin gene in non-O157 E. coli. Standard microbiological techniques identified a total of 137 E. coli isolates. The plasmid was detected by Perfectprep Plasmid Mini preparation kit. These isolates were subjected to disk diffusion assay, and plasmid curing with ethidium bromide treatment. The plasmid containing isolates were subjected to a polymerase chain reaction (PCR) for investigating the presence of plasmid mediated verotoxin gene (VT1 and VT2) in non-O157 E. coli. Among the 137 E. coli isolates, 49 isolates were non-O157 E. coli while 29 (59.1%) isolates were verotoxin producing non-O157 serotypes and 26 non-O157 VTEC isolates possessed plasmids. Certain isolates harboured single sized plasmid while others had multiple plasmids with different size varied from 1.8kb to 7.6kb. A plasmid containing all (100%) the isolates was multidrug-resistant. Eight isolates changed their susceptibility patterns while three isolates were found to lose plasmid after post plasmid curing treatment and the rest of the isolates (15) remained constant. Different PCR sets characterized 3 plasmid-mediated verotoxins producing non-O157 E. coli. This current study demonstrated the occurrence of plasmid mediated verotoxin gene in non-O157 E. coli. To the best of our knowledge, this is the first report in the global literature on plasmid-mediated verotoxin gene in non-O157 E. coli. Timely diagnosis and surveillance of VTEC infections should prioritize to stop or slow down the virulence gene for dissemination by plasmid-mediated gene transfer amongst the same bacteria or other species.

Occurrence and Characterization of Escherichia coli O157 and Other Serotypes in Raw Meat Products in Morocco

2008 ◽  
Vol 71 (10) ◽  
pp. 2082-2086 ◽  
Author(s):  
LUCIANO BENEDUCE ◽  
GIUSEPPE SPANO ◽  
ARI Q. NABI ◽  
FRANCESCO LAMACCHIA ◽  
SALVATORE MASSA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Susceptibility Testing ◽  
Meat Products ◽  
Molecular Techniques ◽  
Escherichia Coli O157 ◽  
Raw Meat ◽  
E Coli ◽  
Meat Samples ◽  
The City

In this study, 100 raw meat samples were collected from 15 local Moroccan butcheries in five different areas of the city of Rabat during a period of 4 months. Overall, 7 of 15 butcheries from three areas of the city yielded strains of Escherichia coli O157. Single isolates from 9 (9%) of 100 raw meat samples were biochemically and serologically confirmed as E. coli O157. Using molecular techniques, two strains were positive for the Shiga toxin, with two additional strains containing an attaching-effacing gene. All potentially virulent serotypes isolated from these meat samples showed distinct pulsed-field gel electrophoresis profiles. Based on antibiotic susceptibility testing, more than 70% of the isolates were resistant to ampicillin and clavulanic acid–amoxicillin. Moreover, one strain was resistant to more than three antibiotics. Our study represents the first survey of E. coli O157 and related serotypes in raw meat products in Morocco.

Antibiotic Resistance Profile of Commensal Escherichia coli Isolated from Broiler Chickens in Qatar

2017 ◽  
Vol 81 (2) ◽  
pp. 302-307 ◽  
Author(s):  
Nahla O. Eltai ◽  
Elmoubasher A. Abdfarag ◽  
Hamad Al-Romaihi ◽  
Eman Wehedy ◽  
Mahmoud H. Mahmoud ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Broiler Chickens ◽  
Susceptibility Testing ◽  
Multidrug Resistant ◽  
Test Method ◽  
Health Concern ◽  
E Coli ◽  
Extended Spectrum ◽  
Stewardship Program

ABSTRACT Antibiotic resistance (AR) is a growing public health concern worldwide, and it is a top health challenge in the 21st century. AR among Enterobacteriaceae is rapidly increasing, especially in third-generation cephalosporins and carbapenems. Further, strains carrying mobilized colistin resistance (mcr) genes 1 and 2 have been isolated from humans, food-producing animals, and the environment. The uncontrolled use of antibiotics in food-producing animals is a major factor in the generation and spread of AR. No studies have been done to evaluate AR in the veterinary sector of Qatar. This study aimed at establishing primary baseline data for the prevalence of AR among food-producing animals in Qatar. Fecal samples (172) were obtained from two broiler farms and one live bird market in Qatar, and 90 commensal Escherichia coli bacteria were isolated and subjected to susceptibility testing against 16 clinically relevant antibiotics by using the E-test method. The results found that 81 (90%) of 90 isolates were resistant to at least one antibiotic, 14 (15.5%) of 90 isolates were colistin resistant, 2 (2.2%) of 90 isolates were extended-spectrum β-lactamase producers, and 2 (2.2%) of 90 isolates were multidrug resistant to four antibiotic classes. Extended-spectrum β-lactamase–producing E. coli and colistin-resistant isolates were confirmed by using double-disc susceptibility testing and PCR, respectively. Such a high prevalence of antibiotic-resistant E. coli could be the result of a long application of antibiotic treatment, and it is an indicator of the antibiotic load in food-producing animals in Qatar. Pathogens carrying AR can be easily transmitted to humans through consumption of undercooked food or noncompliance with hygiene practices, mandating prompt development and implementation of a stewardship program to control and monitor the use of antibiotics in the community and agriculture.

scholarly journals Characterization of Multidrug-Resistant Escherichia coli Isolates from Animals Presenting at a University Veterinary Hospital

2011 ◽  
Vol 77 (20) ◽  
pp. 7104-7112 ◽  
Author(s):  
Maria Karczmarczyk ◽  
Yvonne Abbott ◽  
Ciara Walsh ◽  
Nola Leonard ◽  
Séamus Fanning
Keyword(s):  
Escherichia Coli ◽  
Molecular Mechanisms ◽  
Gene Array ◽  
Multidrug Resistant ◽  
Genetic Determinants ◽  
Gene Encoding ◽  
Content Type ◽  
E Coli ◽  
Class 1

ABSTRACTIn this study, we examined molecular mechanisms associated with multidrug resistance (MDR) in a collection ofEscherichia coliisolates recovered from hospitalized animals in Ireland. PCR and DNA sequencing were used to identify genes associated with resistance. Class 1 integrons were prevalent (94.6%) and contained gene cassettes recognized previously and implicated mainly in resistance to aminoglycosides, β-lactams, and trimethoprim (aadA1,dfrA1-aadA1,dfrA17-aadA5,dfrA12-orfF-aadA2,blaOXA-30-aadA1,aacC1-orf1-orf2-aadA1,dfr7). Class 2 integrons (13.5%) contained thedfrA1-sat1-aadA1gene array. The most frequently occurring phenotypes included resistance to ampicillin (97.3%), chloramphenicol (75.4%), florfenicol (40.5%), gentamicin (54%), neomycin (43.2%), streptomycin (97.3%), sulfonamide (98.6%), and tetracycline (100%). The associated resistance determinants detected includedblaTEM,cat,floR,aadB,aphA1,strA-strB,sul2, andtet(B), respectively. TheblaCTX-M-2gene, encoding an extended-spectrum β-lactamase (ESβL), andblaCMY-2, encoding an AmpC-like enzyme, were identified in 8 and 18 isolates, respectively. The mobility of the resistance genes was demonstrated using conjugation assays with a representative selection of isolates. High-molecular-weight plasmids were found to be responsible for resistance to multiple antimicrobial compounds. The study demonstrated that animal-associated commensalE. coliisolates possess a diverse repertoire of transferable genetic determinants. Emergence of ESβLs and AmpC-like enzymes is particularly significant. To our knowledge, theblaCTX-M-2gene has not previously been reported in Ireland.

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 Characterization of commensal Escherichia coli isolates from slaughtered sheep in Mexico

2021 ◽  
Vol 15 (11) ◽  
pp. 1755-1760
Author(s):  
Jorge Acosta-Dibarrat ◽  
Edgar Enriquez-Gómez ◽  
Martín Talavera-Rojas ◽  
Edgardo Soriano-Vargas ◽  
Armando Navarro ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Multidrug Resistant ◽  
Phylogenetic Group ◽  
Phylogenetic Groups ◽  
E Coli ◽  
New Information ◽  
Antimicrobial Resistance Genes ◽  
Uremic Syndrome

Introduction: Commensal Escherichia coli is defined as bacteria without known virulence factors that could be playing a specific role in some diseases; however, they could be responsible to disseminate antimicrobial resistance genes to other microorganisms. This study aimed to characterize the commensal E. coli isolates obtained from slaughtered sheep in the central region of Mexico. Methodology: Isolates were classified as commensal E. coli when distinctive genes related to diarrheagenic pathotypes (stx1, stx2, eae, bfp, LT, stp, ipaH, and aggR) were discarded by PCR. Identification of serotype, phylogenetic group, and antimicrobial resistance was also performed. Results: A total of 41 isolates were characterized. The phylogenetic groups found were B1 in 37 isolates (90.2%), A in 2 (4.8%), and 1 isolate (2.4%) for C and D groups. Serotypes associated with diarrhea in humans (O104:H2 and O154:NM) and hemolytic uremic syndrome (O8:NM) were detected. Thirty-three isolates (80%) were resistant to ceftazidime, 23 (56%), to tetracycline 8 (19.5%) to ampicillin, and 1 to amikacin. Six isolates (14.6%) were multidrug-resistant. Conclusions: This study provides new information about commensal E. coli in slaughtered sheep, high percentages of resistance to antibiotics, and different profiles of antimicrobial resistance were found, their dissemination constitute a risk factor towards the consuming population.

scholarly journals Characterization of auto-agglutinating and non-typeable uropathogenic Escherichia coli strains

2019 ◽  
Vol 13 (06) ◽  
pp. 465-472
Author(s):  
Ulises Hernández-Chiñas ◽  
Alejandro Pérez-Ramos ◽  
Laura Belmont-Monroy ◽  
María E Chávez-Berrocal ◽  
Edgar González-Villalobos ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Virulence Genes ◽  
Antimicrobial Agents ◽  
Multidrug Resistant ◽  
Uropathogenic Escherichia Coli ◽  
E Coli ◽  
Resistant Strains ◽  
Tract Infections ◽  
Disk Method

Introduction: Uropathogenic Escherichia coli (UPEC) are the main etiological agent of urinary tract infections (UTIs). Association between different serotypes and UTIs is known, however, some strains are incapable to be serotyped. The aim of this work was to study bthe phenotypical and genotypical characteristics of 113 non-typeable (NT) and auto-agglutinating (AA) E. coli strains, isolated from UTIs in children and adults. Methodology: The 113 UPEC strains were analyzed by PCR assays using specific primers to determine their serogroups, fimH, papC, iutA, sat, hlyCA and cnf1, virulence associated genes, and chuA, yjaA and TSPE4.C2 for phylogroup determination. Additionally, the diffusion disk method was performed to evaluate the antimicrobial resistance to 18 antimicrobial agents. Results: Using the PCR assay, 63% (71) of the strains were genotyped showing O25 and O75 as the most common serogroups. The virulence genes fimH (86%) and iutA (74%) were the most prevalent, in relation to the phylogroups the commensal (A and B1) and virulent (B2 and D) showed similar frequencies (P > 0.05). The antimicrobial susceptibility test showed a high percentage (73%) of multidrug-resistant strains. Conclusions: The genotyping allowed identifying the serogroup in many of the strains that could not be typed by traditional serology. The strains carried virulence genes and were multidrug-resistant in both, commensal and virulent phylogroups. Our findings revealed that, in addition to the classical UPEC serogroups, there are pathogenic serogroups not reported yet.

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.

Characterization of multidrug-resistant Escherichia coli isolated from extraintestinal clinical infections in animals

2010 ◽  
Vol 59 (5) ◽  
pp. 592-598 ◽  
Author(s):  
Justine S. Gibson ◽  
Rowland N. Cobbold ◽  
Darren J. Trott
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Susceptibility ◽  
Animal Species ◽  
Antimicrobial Agents ◽  
Multidrug Resistant ◽  
Phylogenetic Group ◽  
Phylogenetic Groups ◽  
Diagnostic Laboratory ◽  
E Coli

Multidrug-resistant (MDR) Escherichia coli causes extraintestinal infections in both humans and animals. This study aimed to determine whether MDR E. coli isolates cultured from extraintestinal infections in several animal species were clonal and crossed host-species boundaries, as suggested by initial characterization of a subset of canine and human isolates, or whether they represented a diverse group of host-specific strains. Isolates were obtained either from The University of Queensland Veterinary Diagnostic Laboratory or from an independent diagnostic laboratory between October 1999 and December 2007. Ninety-six MDR E. coli isolates cultured from extraintestinal clinical infections in 55 animals comprising dogs (n=45), cats (n=5), horses (n=4) and a koala (n=1) were analysed by phylogenetic grouping, antimicrobial susceptibility testing and PFGE. The isolates were cultured from the urinary tract (n=61), reproductive tract (n=11), wounds (n=11), surgical site infections (n=4) and other sites (n=9). Isolates from the same E. coli phylogenetic group with 100 % PFGE similarity and the same antimicrobial susceptibility pattern were considered to be repeat clones and excluded from further analysis. Three of the four E. coli phylogenetic groups (A, n=19; B1, n=8; and D, n=49) were represented. Analysis of PFGE similarity identified clusters of related phylogenetic group A isolates [clonal group (CG) 1] and group D isolates (CG2 and CG3), with the remainder of the isolates demonstrating diversity. The majority of CG2 isolates contained a plasmid-borne AmpC β-lactamase, imparting resistance to cefoxitin and third-generation cephalosporins, and were obtained between 2000 and 2003. CG3 isolates were sensitive to these antimicrobial agents and appeared to replace CG2 isolates as the dominant clones from 2003 to 2007. Apart from several canine and feline isolates that demonstrated clonality, PFGE profiles tended to be divergent across species. Whilst MDR E. coli isolates from extraintestinal infections in different animal species are diverse, some dominant CGs may persist over several years.

Sign in / Sign up

Export Citation Format

Share Document