scholarly journals Molecular and phenotypic typing of enteropathogenic Escherichia coli isolated in childhood acute diarrhea in Abuja, Nigeria

2017 ◽  
Vol 11 (07) ◽  
pp. 527-535 ◽  
Author(s):  
Casmir Ifeanyichukwu Cajetan Ifeanyi ◽  
Nkiruka Florence Ikeneche ◽  
Bassey Enya Bassey ◽  
Stefano Morabito ◽  
Caterina Graziani ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Acute Diarrhea ◽  
Antimicrobial Agents ◽  
Virulence Gene ◽  
Enteropathogenic Escherichia Coli ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Typical Epec ◽  
Marked Resistance

Introduction: Enteropathogenic Escherichia coli (EPEC) causes infectious diarrhea among children in developing countries. However, in Nigeria, due to limited laboratory resources, the genetic diversity of its virulence factors, which include intimin subtypes, remains undefined. Methodology: EPEC isolates from diarrheic children 60 months of age and younger in Abuja, Nigeria, were analyzed. Polymerase chain reaction (PCR) for EPEC virulence gene, Hep-2 cell adherence, and serotyping were performed. EPEC strains were further subtyped by PCR for the identification of intimin subtype genes α (alpha), β (beta), γ1 (gamma-1), and έ (epsilon). Antibiotic resistance and extended-spectrum beta-lactamase (ESBL) production was determined by Clinical and Laboratory Standards Institute guidelines. Results: Overall, 18 (4.5%) out of 400 children with acute diarrhea had EPEC infection.  Typical EPEC (tEPEC) strains were detected in 14 (3.5%), whereas 4 (1.1%) were atypical EPEC (aEPEC). A total of 15 (83.3%) of the EPEC isolated belonged to β intimin subtype gene, while the remaining 3 EPEC isolates possessed the intimin έ subtype. No α and γ intimin subtypes were detected. Traditional EPEC serotypes O114:H14 were detected only in tEPEC strains. Marked resistance to β-lactam agents were observed but no ESBL-producing tEPEC or aEPEC was detected. Conclusions: This is the first report of intimin subtype genes in Abuja, Nigeria. EPEC isolates of diverse serotypes resistant to β-lactam antimicrobial agents were observed. These data will be useful in facilitating the characterization of intimin variants of EPEC and some Shiga toxin-producing E. coli (STEC) in humans and other animal species.

scholarly journals Characterization of typical and atypical enteropathogenic Escherichia coli (EPEC) strains of the classical O55 serogroup by RAPD analysis

1999 ◽  
Vol 30 (4) ◽  
pp. 365-368 ◽  
Author(s):  
Dennys M. Girão ◽  
Sílvia Y. Bando ◽  
Valéria Brígido de C. Girão ◽  
Carlos A. Moreira-Filho ◽  
Sérgio Eduardo L. Fracalanzza ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Rapd Analysis ◽  
Random Amplified Polymorphic Dna ◽  
Epidemiological Studies ◽  
Enteropathogenic Escherichia Coli ◽  
E Coli ◽  
Typical Epec ◽  
Rapd Method ◽  
Atypical Strains

The genetic diversity of 41 typical and atypical enteropathogenic Escherichia coli (EPEC) strains of the serogroup O55 was analyzed by using the random amplified polymorphic DNA (RAPD) method. All typical EPEC O55 strains were grouped in two clusters (A and C) and belonged to the serotype O55:H6, while cluster B included all atypical strains, which were of the serotype O55:H7. The three groups also included non-motile strains. RAPD may be a useful method for epidemiological studies on E. coli O55 infection.

Isolation of atypical enteropathogenic Escherichia coli from children with and without diarrhoea in Delhi and the National Capital Region, India

2010 ◽  
Vol 59 (10) ◽  
pp. 1156-1162 ◽  
Author(s):  
Pankaj Kumar Ghosh ◽  
Arif Ali
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Agents ◽  
Virulence Gene ◽  
Healthy Children ◽  
National Capital Region ◽  
Capital Region ◽  
E Coli ◽  
Atypical Epec ◽  
National Capital ◽  
Typical Epec

A total of 17 typical and atypical enteropathogenic Escherichia coli (EPEC) were isolated from 396 children with and without diarrhoea. Out of 12 EPEC isolates from patients with diarrhoea, 3 (25 %) were atypical EPEC while 9 (75 %) were typical EPEC. It was observed that atypical EPEC strains had colonized the intestines of healthy children and its isolation rates were higher in healthy children than in children with diarrhoea. Interestingly all of the atypical EPEC isolates carried a megaplasmid, mostly comparable with the size of EPEC adherence factor (EAF) encoding gene but no virulence gene was detected in this megaplasmid. Studies also indicated that multidrug resistance EPEC are emerging and all the atypical EPEC strains showed significantly less resistance to all antimicrobial agents used in this study than typical EPEC. This study also supports the opinion that Shiga toxin-producing E. coli does not pose a major threat to human health in India. Subtyping analysis reveals that eae-α1, eae-β2 and eae-λ could be common EPEC subtypes prevalent in children with diarrhoea in Delhi. The present study is believed to be the first report of the detection of atypical EPEC from children without diarrhoea and records of isolation of eae-γ1, eae-γ2 and the rare eae-λ subtype in India. The data also indicated that typical EPEC are a common cause of diarrhoea and atypical EPEC are emerging as colonizers of the intestine of children with and without diarrhoea in Delhi and the National Capital Region, India.

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.

scholarly journals Molecular characterisation of antimicrobial resistance and virulence genes in Escherichia coli strains isolated from diarrhoeic and healthy rabbits in Tunisia

2020 ◽  
Vol 28 (2) ◽  
pp. 81
Author(s):  
Raouia Ben Rhouma ◽  
Ahlem Jouini ◽  
Amira Klibi ◽  
Safa Hamrouni ◽  
Aziza Boubaker ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Virulence Genes ◽  
Antimicrobial Agents ◽  
Antibiotic Resistance Genes ◽  
Virulence Gene ◽  
Diffusion Method ◽  
Class 1 Integron ◽  
E Coli ◽  
Class 1

The purpose of this study was to identify <em>Escherichia coli</em> isolates in diarrhoeic and healthy rabbits in Tunisia and characterise their virulence and antibiotic resistance genes. In the 2014-2015 period, 60 faecal samples from diarrhoeic and healthy rabbits were collected from different breeding farms in Tunisia. Susceptibility to 14 antimicrobial agents was tested by disc diffusion method and the mechanisms of gene resistance were evaluated using polymerase chain reaction and sequencing methods. Forty <em>E. coli</em> isolates were recovered in selective media. High frequency of resistance to tetracycline (95%) was detected, followed by different levels of resistance to sulphonamide (72.5%), streptomycin (62.5%), trimethoprim-sulfamethoxazole (60%), nalidixic acid (32.5%), ampicillin (37.5%) and ticarcillin (35%). <em>E. coli</em> strains were susceptible to cefotaxime, ceftazidime and imipenem. Different variants of bla<sub>TEM</sub>, <em>tet</em>, <em>sul</em> genes were detected in most of the strains resistant to ampicillin, tetracycline and sulphonamide, respectively. The presence of class 1 integron was studied in 29 sulphonamide-resistant <em>E. coli</em> strains from which 15 harboured class 1 integron with four different arrangements of gene cassettes, <em>dfrA17</em>+<em>aadA5</em> (n=9), <em>dfrA1</em> + <em>aadA1</em> (n=4), <em>dfrA12</em> + <em>addA2</em> (n=1), <em>dfrA12</em>+<em>orf</em>+<em>addA2</em> (n=1). The <em>qnrB</em> gene was detected in six strains out of 13 quinolone-resistant <em>E. coli</em> strains. Seventeen <em>E. coli</em> isolates from diarrhoeic rabbits harboured the enteropathogenic eae genes associated with different virulence genes tested (<em>fimA</em>, <em>cnf1</em>, <em>aer</em>), and affiliated to B2 (n=8) and D (n=9) phylogroups. Isolated <em>E. coli</em> strains from healthy rabbit were harbouring <em>fim A</em> and/or <em>cnf1</em> genes and affiliated to A and B1 phylogroups. This study showed that <em>E. coli</em> strains from the intestinal tract of rabbits are resistant to the widely prescribed antibiotics in medicine. Therefore, they constitute a reservoir of antimicrobial-resistant genes, which may play a significant role in the spread of antimicrobial resistance. In addition, the eae virulence gene seemed to be implicated in diarrhoea in breeder rabbits in Tunisia.

scholarly journals Extended-spectrum beta-lactamase (ESBL)-producing and non-ESBL-producing Escherichia coli isolates causing bacteremia in the Netherlands (2014 – 2016) differ in clonal distribution, antimicrobial resistance gene and virulence gene content

2019 ◽  
Author(s):  
Denise van Hout ◽  
Tess D. Verschuuren ◽  
Patricia C.J. Bruijning-Verhagen ◽  
Thijs Bosch ◽  
Anita C. Schürch ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
The Netherlands ◽  
Resistance Gene ◽  
Acquired Resistance ◽  
Virulence Gene ◽  
Beta Lactamase ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum ◽  
Clonal Distribution

ABSTRACTBackgroundKnowledge on the molecular epidemiology of Escherichia coli causing E. coli bacteremia (ECB) in the Netherlands is mostly based on extended-spectrum beta-lactamase-producing E. coli (ESBL-Ec). We determined differences in clonality and resistance and virulence gene (VG) content between non-ESBL-producing E. coli (non-ESBL-Ec) and ESBL-Ec blood isolates with different epidemiological characteristics.Materials/methodsA random selection of non-ESBL-Ec isolates as well as all available ESBL-Ec blood isolates was obtained from two Dutch hospitals between 2014 and 2016. Whole genome sequencing was performed to infer sequence types (STs), serotypes, acquired antibiotic resistance genes and VG scores, based on presence of 49 predefined putative pathogenic VG.ResultsST73 was most prevalent among the 212 non-ESBL-Ec (N=26, 12.3%) and ST131 among the 69 ESBL-Ec (N=30, 43.5%). Prevalence of ST131 among non-ESBL-Ec was 10.4% (N=22, P value < 0.001 compared to ESBL-Ec). O25:H4 was the most common serotype in both non-ESBL-Ec and ESBL-Ec. Median acquired resistance gene counts were 1 (IQR 1 – 6) and 7 (IQR 4 – 9) for non-ESBL-Ec and ESBL-Ec, respectively (P value < 0.001). Among non-ESBL-Ec, acquired resistance gene count was highest among blood isolates from a primary gastro-intestinal focus (median 4, IQR 1 – 8). Median VG scores were 13 (IQR 9 – 20) and 12 (IQR 8 – 14) for non-ESBL-Ec and ESBL-Ec, respectively (P value = 0.002). VG scores among non-ESBL-Ec from a primary urinary focus (median 15, IQR 11 – 21) were higher compared to non-ESBL-Ec from a primary gastro-intestinal (median 10, IQR 6 – 13) or hepatic-biliary focus (median 11, IQR 5 – 18) (P values = 0.007 and 0.036, respectively). VG content varied between different E. coli STs.ConclusionsNon-ESBL-Ec and ESBL-Ec blood isolates from two Dutch hospitals differed in clonal distribution, resistance gene and VG content. Also, resistance gene and VG content differed between non-ESBL-Ec from different primary foci of ECB.

scholarly journals Extended-Spectrum Beta-Lactamase-Producing Escherichia coli Meningitis and Cerebral Abscess in a Neonate: Therapeutic Challenge

2019 ◽  
Vol 2019 ◽  
pp. 1-5
Author(s):  
Surasak Puvabanditsin ◽  
Marianne Jacob ◽  
Maaz Jalil ◽  
Samhita Bhattarai ◽  
Qaiser Patel ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Agents ◽  
Cerebral Abscess ◽  
Term Neonate ◽  
Resistant Bacteria ◽  
Beta Lactamase ◽  
Therapeutic Challenge ◽  
Extended Spectrum Beta Lactamase ◽  
E Coli ◽  
Extended Spectrum

We report a case of a 12-day-old term neonate with extended-spectrum beta-lactamase (ESBL) producing Escherichia coli (E. coli) meningitis and cerebral abscess. The patient received a 7-day course of antibiotics just few days prior to the infection. The incidence of infections from ESBL-producing E. coli is increasingly emerging. Antimicrobial agents must be vigilantly utilized to prevent the new highly resistant bacteria.

scholarly journals Phenotypic and Genotypic Characterization of Escherichia coli Isolated from Untreated Surface Waters

2013 ◽  
Vol 7 (1) ◽  
pp. 9-19 ◽  
Author(s):  
Kristopher J Janezic ◽  
Blake Ferry ◽  
Eric W Hendricks ◽  
Brian A Janiga ◽  
Tiffany Johnson ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Surface Waters ◽  
Antimicrobial Agents ◽  
Genotypic Variation ◽  
Virulence Gene ◽  
Genotypic Diversity ◽  
Housekeeping Gene ◽  
Polymyxin B ◽  
Fecal Coliforms ◽  
E Coli

A common member of the intestinal microbiota in humans and animals isEscherichia coli. Based on the presence of virulence factors,E. colican be potentially pathogenic. The focus of this study was to isolateE. colifrom untreated surface waters (37 sites) in Illinois and Missouri and determine phenotypic and genotypic diversity among isolates. Water samples positive for fecal coliforms based on the Colisure®test were streaked directly onto Eosin Methylene Blue (EMB) agar (37°C) or transferred to EC broth (44.5°C). EC broth cultures producing gas were then streaked onto EMB agar. Forty-five isolates were identified asE. coliusing API 20E and Enterotube II identification systems, and some phenotypic variation was observed in metabolism and fermentation. Antibiotic susceptibility of each isolate was also determined using the Kirby-Bauer Method. Differential responses to 10 antimicrobial agents were seen with 7, 16, 2, and 9 of the isolates resistant to ampicillin, cephalothin, tetracycline, and triple sulfonamide, respectively. All of the isolates were susceptible or intermediate to amoxicillin, ciprofloxacin, polymyxin B, gentamicin, imipenem, and nalidixic acid. Genotypic variation was assessed through multiplex Polymerase Chain Reaction for four virulence genes (stx1andstx2[shiga toxin],eaeA[intimin]; andhlyA[enterohemolysin]) and one housekeeping gene (uidA[β-D-glucuronidase]). Genotypic variation was observed with two of the isolates possessing the virulence gene (eaeA) for intimin. These findings increase our understanding of the diversity ofE. coliin the environment which will ultimately help in the assessment of this organism and its role in public health.

Detection of Multiple-Antimicrobial Resistance and Characterization of the Implicated Genes in Escherichia coli Isolates from Foods of Animal Origin in Tunis

2009 ◽  
Vol 72 (5) ◽  
pp. 1082-1088 ◽  
Author(s):  
AHLEM JOUINI ◽  
KARIM BEN SLAMA ◽  
YOLANDA SÁENZ ◽  
NAOUEL KLIBI ◽  
DANIELA COSTA ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Antimicrobial Agents ◽  
Food Samples ◽  
Animal Origin ◽  
E Coli ◽  
Antimicrobial Resistance Genes ◽  
Class 1 ◽  
Agar Dilution

Phenotypic and genotypic characterization of antimicrobial resistance was conducted for 98 Escherichia coli isolates recovered from 40 food samples of animal origin (poultry, sheep, beef, fish, and others) obtained in supermarkets and local butcheries in Tunis during 2004 and 2005. Susceptibility to 15 antimicrobial agents was tested by disk diffusion and agar dilution methods, the mechanisms of resistance were evaluated using PCR and sequencing methods, and the clonal relationship among isolates was evaluated using pulsed-field gel electrophoresis. High resistance was detected to tetracycline, sulphonamides, nalidixic acid, ampicillin, streptomycin, and trimethoprim-sulfamethoxazole (29 to 43% of isolates), but all isolates were susceptible to cefotaxime, ceftazidime, cefoxitin, azthreonam, and amikacin. One-third of the isolates had multiresistant phenotypes (resistance to at least five different families of antimicrobial agents). Different variants of blaTEM, tet, sul, dfrA, aadA, and aac(3) genes were detected in most of the strains resistant to ampicillin, tetracycline, sulphonamide, trimethoprim, streptomycin, and gentamicin, respectively. The presence of class 1 and class 2 integrons was studied in 15 sulphonamide-resistant unrelated E. coli strains, and 14 of these strains harbored class 1 integrons with five different arrangements of gene cassettes, and a class 2 integron with the dfrA1 + sat + aadA1 arrangement was found in one strain. This study revealed the high diversity of antimicrobial resistance genes, some of them included in integrons, in E. coli isolates of food origin.

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