scholarly journals Molecular Characterization of β-Lactamase Producing Genes and Integrons in Diarrheagenic Escherichia Coli From Diarrheal Children Less Than Five Years of Age in Ouagadougou, Burkina Faso.

2021 ◽  
Author(s):  
Rene Dembele ◽  
Wendpoulomdé A.D. Kaboré ◽  
Issiaka Soulama ◽  
Oumar Traoré ◽  
Nafissatou Ouédraogo ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Molecular Characterization ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Control Measures ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Disk Diffusion Method ◽  
E Coli

Abstract Background The aim of this study was to determine the resistance of diarrheagenic Escherichia coli strains to β-lactams antibiotics and to perform the molecular characterization of Extended Spectrum β-lactamases (ESBL) and integrons genes. Methods This study was carried out from August 2013 to October 2015 and involved 31 DEC strains isolated from diarrheal stools samples collected from children less than five years of age. The identification and characterization of DEC strains was done through the standard biochemical tests those were confirmed using API 20E and Polymerase Chain Reaction (PCR). The determination of antimicrobial resistance was realized by the disk diffusion method then an amplification of the β-lactamase resistance genes and integrons by PCR was done. Results Out of the 419 E. coli strains identified, 31 isolates (7.4%) harbored the DEC virulence genes. From these DEC, 21 (67.7%) were ESBL-producing E. coli. Susceptibility to ESBL-producing E. coli showed that the majority of isolates were highly resistant to amoxicillin (77.4%), amoxicillin clavulanic acid (77.4%) and piperacillin (64.5%). The following antibiotic resistance genes and integron were identified from the 31 DEC isolates: blaTEM (6.5%), blaSHV (19.4%), blaOXA (38.7%) blaCTX−M (9.7%), Int1 (58.1%) and Int3 (19.4%). No class 2 integrons (Int2) was characterized. Conclusions Because of the high prevalence of multidrug-resistant ESBL organisms found in this study among pediatric patients, there is a need of stringent pediatric infection control measures.

Characterization of Antibiotic Resistance E. Coli and Antibiotic Resistance Genes in Aquatic Environment of Taihu Lake, China

2013 ◽  
Vol 295-298 ◽  
pp. 630-634 ◽  
Author(s):  
Ni Ni Han ◽  
Song He Zhang ◽  
Pei Fang Wang ◽  
Chao Wang
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Surface Water ◽  
Resistance Genes ◽  
Taihu Lake ◽  
Antibiotic Resistance Genes ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Realtime Pcr

The aims of this study are to evaluate multiple antibiotic resistant Escherichia coli isolated from surface water and to investigate the presence and distribution antibiotic resistance genes (ARGs) in sediments of Taihu Lake. The results show that the presentence of four ARGs concentrations in the sediments of the lake was in sequence: strB>qnrB>strA>qnrS, as determined by realtime-PCR technique. The southwest and east areas of Taihu Lake were polluted seriously than other areas from all kinds of antibiotics. The screening Escherichia coli had a higher resistance to streptomycin, tetracycline and ampicillin than other four antibiotics, and had a lowest resistance to levofloxacin.

scholarly journals Prevalence of class 1 and 2 integrons among the multidrug resistant uropathogenic strains of Escherichia coli

2017 ◽  
Vol 9 (1) ◽  
pp. 49-54 ◽  
Author(s):  
Haddadi Azam ◽  
Somayeh Mikaili Ghezeljeh ◽  
Shavandi Mahmoud
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Multidrug Resistant ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
Integrase Gene ◽  
Class 1 ◽  
Tract Infections

Abstract Background Multidrug resistance is a serious problem in the treatment of urinary tract infections. Horizontal gene transfer, directed by strong selective pressure of antibiotics, has resulted in the widespread distribution of multiple antibiotic resistance genes. The dissemination of resistance genes is enhanced when they are trapped in integrons. Objectives To determine the prevalence of integrons among multidrug resistant Escherichia coli strains collected from regional hospitals and private clinical laboratories in Alborz province. Methods The susceptibility of 111 clinical Escherichia coli isolates was tested using a Kirby–Bauer disk diffusion method for common antibiotics. Isolates were screened for the production of extended spectrum β-lactamases (ESBLs) using a double disk synergy test. The existence of integrons was confirmed by amplification of the integrase gene and their class determined via analysis of PCR products by PCR-RFLP. Results Isolates showed the highest resistance to amoxicillin. Nitrofurantoin, amikacin, and ceftizoxime were the most effective antibiotics in vitro. Eighty-eight isolates of 111 (79%) were resistant to more than three unrelated drugs. We found 30% of the multidrug resistant isolates harbor integrons. Class 1 and 2 integrons were detected in 25 and 1 isolates, respectively. ESBL screening of strains showed 45 isolates (40%) were positive; 22% of the ESBL-positive isolates carried class 1 integrons and the frequency of MDR in ESBLpositive isolates was 93%. Conclusion The existence of integrons in only 29.5% of multidrug resistant isolates showed that besides integrons, antibiotic resistance genes were probably carried on other transferable elements lacking integrons, such as transposons or plasmids.

scholarly journals Molecular Characterization of Carbapenemase-Producing Escherichia coli and Salmonella in children with diarrhea in rural Burkina Faso.

2020 ◽  
Author(s):  
Rene DEMBELE ◽  
Ali Konaté ◽  
Issiaka Soulama ◽  
Wendpoulomdé A. D. Kaboré ◽  
Assèta Kagambèga ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Burkina Faso ◽  
Treatment Options ◽  
Diffusion Method ◽  
Bacterial Strains ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Klebsiella Pneumoniae Carbapenemase ◽  
Encoding Genes

Abstract Background In recent years, Carbapenemase-producing Enterobacteriaceae (CPE) resistance to antibiotics has dramatically increased leading to limitations of their treatment options. In the present study, we investigated the occurrence of carbapenemase-producing Escherichia coli and Salmonella in rural Burkina Faso, using bacterial strains obtained from previous studies. Results Diarrheagenic Escherichia coli (DEC) strains was identified using 16-plex Polymerase Chain Reaction (PCR), whereas antibiotic susceptibility was realized using the disk diffusion method. Furthermore, multiplex PCR assays were used to characterize bla KPC, bla VIM and bla IMP genes in carbapenemase-producing E . coli and Salmonella . The study highlighted high resistance rates of the identified bacteria to common antibiotics. Likewise, two strains of E . coli were imipenem resistant with Carbapenemase-encoding genes. The genes detected were Klebsiella pneumoniae carbapenemase (KPC), Verona integrin-encoded metallo-β-lactamase (VIM) and Imipenemase (IMP-2) reaching a rate of 40% each. However, no Carbapenemase-encoding genes were detected in Salmonella isolates. Conclusions This study showed that for a real-time infection control and prompt application of antimicrobial chemotherapy, characterization of carbapenemase-producing Enterobacteriaceae in patients is crucial.

scholarly journals Identification of Extended-Spectrumβ-LactamasesEscherichia coliStrains Isolated from Market Garden Products and Irrigation Water in Benin

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Wassiyath Moussé ◽  
Haziz Sina ◽  
Farid Baba-Moussa ◽  
Pacôme A. Noumavo ◽  
Nadège A. Agbodjato ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Susceptibility ◽  
Irrigation Water ◽  
Food Habits ◽  
Diffusion Method ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Pcr Method ◽  
Horticultural Products

The present study aimed at biochemical and molecular characterization ofEscherichia colistrains isolated from horticultural products and irrigation water of Cotonou. The samples were collected from 12 market gardeners of 4 different sites. Rapid’E. colimedium was used for identification ofE. colistrains and the antimicrobial susceptibility was performed by the agar disk diffusion method. Theβ-lactamases production was sought by the liquid acidimetric method. The genes coding forβ-lactamases and toxins were identified by PCR method. The results revealed that about 34.95% of the analyzed samples were contaminated byE. coli. Cabbages were the most contaminated byE. coli(28.26%) in dry season. All isolated strains were resistant to amoxicillin. The penicillinase producingE. colicarriedblaTEM(67.50%),blaSHV(10%), andblaCTX-M(22.50%) genes. The study revealed that the resistance genes such as SLTI (35.71%), SLTII (35.71%), ETEC (7.15%), and VTEC (21.43%) were carried. Openly to the found results and considering the importance of horticultural products in Beninese food habits, it is important to put several strategies aiming at a sanitary security by surveillance and sensitization of all the actors on the risks of some practices.

scholarly journals Antibiogram of Escherichia coli Isolated from Avian Colibacillosis in Chitwan District of Nepal

2020 ◽  
Vol 8 (1) ◽  
pp. 52-60 ◽  
Author(s):  
D.B. Thapa ◽  
A. Chapagain
Keyword(s):  
Escherichia Coli ◽  
Cross Sectional Study ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Isolation And Identification ◽  
Disk Diffusion Method ◽  
Cross Sectional ◽  
E Coli ◽  
Mar Index ◽  
Avian Disease

A cross-sectional study was conducted at National Avian Disease Investigation Laboratory, Chitwan to determine antibiogram of Escherichia coli isolated from avian colibacillosis cases of broilers and layers in Chitwan. One hundred and sixty (95 from broilers and 65 from layers) liver samples were collected aseptically during postmortem. Samples were taken purposively from dead birds showing lesions perihepatitis, pericarditis, air-saculitis, omphalitis and egg peritonitis. Isolation and identification were made by examination of cultural characteristics of E. coli in MacConkey’s agar, Eosin methylene blue (EMB) agar, Gram’s staining and biochemical tests. Antibiogram of identified E. coli isolate was evaluated against six antibiotics of six different groups by disk diffusion method following CLSI guidelines. One hundred and three E. coliisolates (73 from broilers and 30 from layers) were isolated from one hundred and sixty samples. Highest resistance was observed against Ampicillin (100%) followed by Co-trimoxazole (86.40%), Doxycycline (46.60%), Levofloxacin (45.63%), Nitrofurantoin (26.21%) and Amikacin (10.68%). Nearly about all (96.12%) isolates from 103 isolated E. coli isolates showed multidrugs resistance to two or more than two antimicrobials. All multidrug resistance isolates showed 16 different patterns with each isolate being resistance to at least two drugs. The multiple antibiotic resistance indexing ranged from 0.2 to 0.8 and proportion of isolates with MAR index greater than 0.2 was 96.12%. Int. J. Appl. Sci. Biotechnol. Vol 8(1): 52-60    

scholarly journals Prevalence of Resistance to Quinolone and Fluoroquinolone Antibiotics and Screening of qnr Genes Among Escherichia coli Isolates From Urinary Tract Infection

2017 ◽  
Vol 5 (4) ◽  
pp. 100-105 ◽  
Author(s):  
Mohadese Amiri ◽  
Maziar Jajarmi ◽  
Reza Ghanbarpour
Keyword(s):  
Escherichia Coli ◽  
Urinary Tract ◽  
Nalidixic Acid ◽  
Diffusion Method ◽  
Biochemical Tests ◽  
Phylogenetic Groups ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Fluoroquinolone Antibiotics ◽  
Tract Infections

Background: Antibiotic resistance (AR) is an important challenge in prevention, treatment and control of infectious diseases and is a public health threat for human. Escherichia coli strains are the major causes of urinary tract infections (UTIs). Objective: This research aimed to determine prevalence of resistance to quinolone and fluoroquinolone antibiotics and screen qnr genes among E. coli isolates from UTIs. Materials and Methods: A total of 105 E. coli isolates were obtained from UTI cases in Bojnord city (northeast of Iran) and confirmed by biochemical tests. All strains were studied to determine their resistance to 3 antibiotics including ciprofloxacin, nalidixic acid, and levofloxacin via disk diffusion method. Moreover, the frequency of qnrA, qnrB and qnrS genes and phylogroups was studied by conventional Polymerase chain reaction (PCR). Results: In this study, prevalence of phenotypic AR to ciprofloxacin, nalidixic acid and levofloxacin was 47.6%, 44.8% and 38.1%, respectively. Three isolates were positive for qnrS and 1 isolate was positive for qnrA. Seven phylogenetic groups were also identified as follows: 18% A0, 6.7% A1, 7.6% B1, 1.9% B22, 23.8% B23, 15.3% D1 and 26.7% D2. Conclusion: Prevalence of qnr genes was very low; thus, other types of qnr and plasmid-mediated quinolone resistance genes were probably responsible for the resistance. Phenotypic AR to the 3 antibiotics was found in approximately half of E. coli strains. It is strongly recommended that antibiogram tests should be done before prescribing the ciprofloxacin, nalidixic acid and levofloxacin for UTIs.

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.

Prevalence and Molecular Characterization of Antimicrobial Resistance in Escherichia coli Isolated from Raw Milk and Raw Milk Cheese in Egypt

2018 ◽  
Vol 81 (2) ◽  
pp. 226-232 ◽  
Author(s):  
Rabee A. Ombarak ◽  
Atsushi Hinenoya ◽  
Abdel-Rahman M. Elbagory ◽  
Shinji Yamasaki
Keyword(s):  
Escherichia Coli ◽  
Antimicrobial Resistance ◽  
Resistance Genes ◽  
Raw Milk ◽  
Diffusion Method ◽  
Sequencing Analysis ◽  
Aminoglycoside Resistance ◽  
Disk Diffusion Method ◽  
E Coli ◽  
Extended Spectrum

ABSTRACT The goal of this study was to examine antimicrobial resistance and characterize the implicated genes in 222 isolates of Escherichia coli from 187 samples of raw milk and the two most popular cheeses in Egypt. E. coli isolates were tested for susceptibility to 12 antimicrobials by a disk diffusion method. Among the 222 E. coli isolates, 66 (29.7%) were resistant to one or more antimicrobials, and half of these resistant isolates showed a multidrug resistance phenotype (resistance to at least three different drug classes). The resistance traits were observed to tetracycline (27.5%), ampicillin (18.9%), streptomycin (18.5%), sulfamethoxazole-trimethoprim (11.3%), cefotaxime (4.5%), kanamycin (4.1%), ceftazidime (3.6%), chloramphenicol (2.3%), nalidixic acid (1.8%), and ciprofloxacin (1.4%). No resistance to fosfomycin and imipenem was observed. Tetracycline resistance genes tetA, tetB, and tetD were detected in 53 isolates, 9 isolates, and 1 isolate, respectively, but tetC was not detected. Aminoglycoside resistance genes strA, strB, aadA, and aphA1 were detected in 41, 41, 11, and 9 isolates, respectively. Sulfonamide resistance genes sul1, sul2, and sul3 were detected in 7, 25, and 3 isolates, respectively. Of 42 ampicillin-resistant isolates, blaTEM, blaCTX-M, and blaSHV were detected in 40, 9, and 3 isolates, respectively, and 10 (23.8%) ampicillin-resistant isolates were found to produce extended-spectrum β-lactamase. Each bla gene of extended-spectrum β-lactamase–producing E. coli was further subtyped to be blaCTX-M-15, blaCTX-M-104, blaTEM-1, and blaSHV-12. The class 1 integron was also detected in 28 resistant isolates, and three different patterns were obtained by PCR-restriction fragment length polymorphism. Sequencing analysis of the variable region revealed that four isolates had dfrA12/orfF/aadA2, two had aadA22, and one had dfrA1/aadA1. These data suggest that antimicrobial-resistant E. coli are widely distributed in the milk production and processing environment in Egypt and may play a role in dissemination of antimicrobial resistance to other pathogenic and commensal bacteria.

scholarly journals Phenotypic and genotypic characterization of antibiotic-resistant in Escherichia coli isolates from patients with diarrhea

2019 ◽  
Author(s):  
Mojtaba Bonyadian ◽  
Sara Barati ◽  
Mohammad Reza Mahzounieh
Keyword(s):  
Escherichia Coli ◽  
Antibiotic Resistance ◽  
Resistance Genes ◽  
Antibiotic Resistance Genes ◽  
Diffusion Method ◽  
Main Concern ◽  
Antibiotic Resistant ◽  
E Coli ◽  
Resis Tance ◽  
Stool Samples

Background and Objectives: Escherichia coli is a common enteric pathogen of human and livevestock. Antibiotic resis- tance is the main concern of public health. The aim of this study was to detect this bacterium in stool samples of diarrheal patients and identify the phenotypic and genotypic characterizations of antibiotic-resistant isolates such as dfrA1, sul1, citm, tetA, qnr, aac(3)-IV in Shahrekord. Materials and Methods: Two hundred fifty diarrheal stool samples from patients were collected. Microbiological and biochemical examinations were done to detect E. coli. Phenotypic and genotypic antibiotic resistance of the isolates were determined using dick diffusion method and polymerase chain reaction (PCR), respectively. Results: Among 114 E. coli isolates, the least resistance was for gentamicin (0%) and the most resistance was for trimetho- prim (79.8%). The resistance to sulfamethoxazole, ciprofloxacin, ampicillin, and tetracycline were 71.05%, 10.5%, 52.63%, and 3.5% respectively. The results of PCR assay revealed that 10 isolates contain sul1, 49 isolates harbor citm, 8 isolates tetA, 36 isolates dfrA1, 11 isolates qnr genes but there was no isolate with aac(3)-IV gene. In comparison between phenotypic and genotypic of the isolates revealed that citm, tetA, dfrA1, qnr, sul1, aac(3)-IV genes covered 42.98%, 7.01%, 31.57%, 9.64%, 8.7%, 0% of the antibiotic resistance, respectively. Conclusion: Our results revealed that all isolates harbor one or more antibiotic resistance genes and that the PCR is a fast practical and appropriate method to determine the presence of antibiotic resistance genes.

scholarly journals Isolation and Molecular Characterization of Shigatoxigenic O157 and Non-O157 Escherichia coli in Raw Milk Marketed in Chittagong, Bangladesh

2021 ◽  
Author(s):  
Md. Kauser-Ul Alam ◽  
Nazmul Sarwar ◽  
Shireen Akther ◽  
Monsur Ahmad ◽  
Paritosh Kumar Biswas
Keyword(s):  
Escherichia Coli ◽  
Molecular Characterization ◽  
Raw Milk ◽  
Selective Medium ◽  
Enterohemorrhagic Escherichia Coli ◽  
Macconkey Agar ◽  
Biochemical Tests ◽  
E Coli ◽  
Milk Samples

Background: Quality and microbial safety of milk is demanding day by day as it is considered as a host for pathogenic and spoilage microorganisms. In this study, isolation and molecular characterization of shigatoxigenic O157 and non-O157 Escherichia coli in raw milk marketed in Chittagong, Bangladesh were done on 186 raw milk samples in Bangladesh. Methods: MacConkey agar was initially used to screen for the presence of E. coli and the suspected growth as evidenced by large pink colonies on MacConkey agar. Finally the organism was verified by plating through Eosin Methylene Blue (EMB) agar (a selective medium for E. coli where it produces metallic sheen) and applying standard biochemical tests for E. coli. The presence of virulent genes, Shiga-like toxin (stx1 and stx2), Intimin (eaeA), O157 antigen rfbE and Enterohemorrhagic Escherichia coli (EHEC) Hemolysin (EHEC) hlyA in the contaminating E. coli population was determined by polymerase chain reaction (PCR) run on a thermocycler (Applied Biosystem, 2720 thermal cycler, Singapore). Result: Among the raw milk samples, 33 samples were identified as E.coli positive and among the isolates, 6 (18.18%) were identified as possible EHEC O157 and rest of the isolates (81.82%) were considered as probable non EHEC O157. About, 3.23% (186 samples) EHEC O157 was isolated from raw milk samples. Then all the 33 isolates were taken under PCR assay for the identification of five virulent genes Stx1, Stx2, eaeA, rfbE and hlyA. No virulent genes were found in non- EHEC O157 isolates, but 4 stx2 (66.67%) and 1 hlyA (16.67%) gene were observed in another 4 EHEC O157 isolates out of 6, but one isolates contained the both genes and hence the prevalence of STEC was 2.15% in raw milk. Result indicated poor hygienic standard of raw milk from uncontrolled environments and the increased public health risk of those consuming raw milk from such uncontrolled sources.

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