Detection of New Delhi Metallo b Lactamase gene in Uropathogenic E. coli

The rapid dissemination of antibiotic resistant E. coli is now a worldwide problem. In this study, a total of twenty E. coli obtained from stool were selected to determine resistance to beta lactam antibiotics. Beta–Lactamase are enzymes produced by bacteria that provide multi resistance to beta lactam antibiotics such as penicillin, cephalosporin, cephamycin and carbapenems. Of these isolates (n = 20), 35% were found resistant to Amoxicillin Clavulanate, 5% to Imipenem, 50% to Ceftriaxone and 75% to Ampicillin. PCR amplification confirmed the presence of the New Delhi beta-lactamase gene (blaNDM) in one isolate (5%, n=20). Plasmids of variable sizes were found in all the isolates. Beta lactam antibiotics are now commonly used for the treatment of disease. Resistance of 50% of the isolates to Ceftriaxone is alarming as this indicates that an alternative drug may soon need to replace this antibiotic for successful treatment. The finding of this study is also of public health concern as plasmids were found in most isolates and these mobile genetic elements can be transferred among clinical bacteria, thereby disseminating antibiotic resistance further limiting treatment options. Bangladesh J Microbiol, Volume 36 Number 1 June 2019, pp 29-33

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Beta‐lactamase‐producing Escherichia coli in Bangladesh: Their phenotypic and molecular characteristics

Dhaka University Journal of Biological Sciences ◽

10.3329/dujbs.v28i1.46494 ◽

2019 ◽

Vol 28 (1) ◽

pp. 71-81

Author(s):

Sunjukta Ahsan ◽

Riajul Islam

Keyword(s):

Health Concern ◽

Resistance Pattern ◽

Molecular Characteristics ◽

New Delhi ◽

Beta Lactamase ◽

Beta Lactam ◽

Class 1 Integron ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Class 1

The emergence and rapid dissemination of beta-lactamase-producing E. coli is now a worldwide problem. A total of 45 E. coli obtained from clinical specimens from a medical service centre in Dhaka were selected for this study. Test E. coli exhibited variable resistance to 3rd (71.7 - 97.8%, n = 48) and 4th (78%, n = 48) generation beta-lactam antibiotics, with 72% sensitivity to Carbapenem. Analysis of co-resistance indicated that 33.3% of E. coli (n = 48) were co-resistant to beta-lactams and ciprofloxacin. ESBL producers were predominant comprising of 84.7% E. coli. Among them, 22.7% contained blaTEM, 24.2% contained blaCTX-M, 4.3% contained blaSHV and 9.1% contained blaOXA-1 genes. A total of 25.75% isolates were metallo beta-lactamase producers. Of these, 1.5% of E. coli strains contained New Delhi metallo beta-lactamase gene and 6% contained AmpC gene. Multiple beta-lactamase genes were detected in some test isolates; 6.7% isolates contained 4, 20% contained 3 and 73.3% contained 2 beta lactamase genes. Fifty per cent of the E. coli contained plasmids of variable sizes. In addition, a total of 39% of the E. coli contained Class 1 integron. The increasing trend in beta-lactam resistance is of public health concern as it limits treatment regime and indicates to the need of continuous monitoring of resistance pattern. Dhaka Univ. J. Biol. Sci. 28(1): 71-81, 2019 (January)

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Multiplication of ampC upon Exposure to a Beta-Lactam Antibiotic Results in a Transferable Transposon in Escherichia coli

International Journal of Molecular Sciences ◽

10.3390/ijms22179230 ◽

2021 ◽

Vol 22 (17) ◽

pp. 9230

Author(s):

Tania S. Darphorn ◽

Yuanqing Hu ◽

Belinda B. Koenders-van Sintanneland ◽

Stanley Brul ◽

Benno H. ter Kuile

Keyword(s):

Escherichia Coli ◽

Antimicrobial Resistance ◽

Beta Lactamase ◽

Beta Lactam ◽

Variable Regions ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Antimicrobial Resistance Genes ◽

Lactam Antibiotic ◽

Lactamase Gene

Plasmids play a crucial role in spreading antimicrobial resistance genes. Plasmids have many ways to incorporate various genes. By inducing amoxicillin resistance in Escherichia coli, followed by horizontal gene transfer experiments and sequencing, we show that the chromosomal beta-lactamase gene ampC is multiplied and results in an 8–13 kb contig. This contig is comparable to a transposon, showing similarities to variable regions found in environmental plasmids, and can be transferred between E. coli cells. As in eight out of nine replicate strains an almost completely identical transposon was isolated, we conclude that this process is under strict control by the cell. The single transposon that differed was shortened at both ends, but otherwise identical. The outcome of this study indicates that as a result of exposure to beta-lactam antibiotics, E. coli can form a transposon containing ampC that can subsequently be integrated into plasmids or genomes. This observation offers an explanation for the large diversity of genes in plasmids found in nature and proposes mechanisms by which the dynamics of plasmids are maintained.

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First Report of New Delhi Metallo-β-Lactamase-1 (NDM-1) Among Escherichia Coli Strains Isolated from Leukemia Patients in Iran

10.21203/rs.3.rs-42141/v1 ◽

2020 ◽

Author(s):

Mahdane Roshani ◽

Alireza Goodarzi ◽

Sanaz Dehbashi ◽

Farhad Afrasiabi ◽

Hossein Goudarzi ◽

...

Keyword(s):

Escherichia Coli ◽

Pcr Amplification ◽

Opportunistic Pathogen ◽

New Delhi ◽

Beta Lactam ◽

Case Presentation ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Medical Centers ◽

Life Threatening

Abstract Background: Escherichia coli has appeared as an important opportunistic pathogen responsible for nosocomial infections in Patients with immunodeficiency particularly in leukemia patients. New Delhi metallo-β-lactamase (NDM-1) is an enzyme that class of beta-lactam antibiotics and is used in treatment of multi-drug resistant (MDR) infections. Case Presentation: In this study, 80 isolates of Escherichia coli and Klebsiella pneumoniae collected over the course of two years from two medical centers of Tehran, Iran. Production of carbapenemase was detected of the isolates using MHT. New Delhi metallo-beta-lactamase 1 (blaNDM-1) genes were detected by polymerase chain reaction (PCR) ampliﬁcation with speciﬁc primers 2 blaNDM-1 producing E.coli strains were isolated from 2 leukemia of patients, The isolates were resistant to carbapenems (imipenem, meropenem),two isolates were positive for carbapenemase production by Modified Hodge test.Conclusions: The emerging of NDM-1 producing E. coli is a threat for leukemia patients in Oncology and hematology departments. We concluded that the incidence of MDR pathogens increased amang patients with leukemia and is life threatening.

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Characterization of Multidrug Resistant Extended-Spectrum Beta-Lactamase-ProducingEscherichia coliamong Uropathogens of Pediatrics in North of Iran

BioMed Research International ◽

10.1155/2015/309478 ◽

2015 ◽

Vol 2015 ◽

pp. 1-7 ◽

Cited By ~ 24

Author(s):

Mohammad Sadegh Rezai ◽

Ebrahim Salehifar ◽

Alireza Rafiei ◽

Taimour Langaee ◽

Mohammadreza Rafati ◽

...

Keyword(s):

Antimicrobial Resistance ◽

Multidrug Resistant ◽

Beta Lactamase ◽

Beta Lactam ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Extended Spectrum ◽

Extended Spectrum Beta Lactamases ◽

North Of Iran

Escherichia coliremains as one of the most important bacteria causing infections in pediatrics and producing extended-spectrum beta-lactamases (ESBLs) making them resistant to beta-lactam antibiotics. In this study we aimed to genotype ESBL-producingE. coliisolates from pediatric patients for ESBL genes and determine their association with antimicrobial resistance. One hundred of theE. coliisolates were initially considered ESBL producing based on their MIC results. These isolates were then tested by polymerase chain reaction (PCR) for the presence or absence ofCTX,TEM,SHV,GES, andVEBbeta-lactamase genes. About 30.5% of isolatedE. coliwas ESBL-producing strain. TheTEMgene was the most prevalent (49%) followed bySHV(44%),CTX(28%),VEB(8%), andGES(0%) genes. The ESBL-producingE. coliisolates were susceptible to carbapenems (66%) and amikacin (58%) and showed high resistance to cefixime (99%), colistin (82%), and ciprofloxacin (76%). In conclusion, carbapenems were the most effective antibiotics against ESBl-producingE. coliin urinary tract infection in North of Iran. The most prevalent gene is the TEM-type, but the other resistant genes and their antimicrobial resistance are on the rise.

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ESBL Producers in Gram Negative Isolates from Clinical Samples

Journal of Pure and Applied Microbiology ◽

10.22207/jpam.14.3.42 ◽

2020 ◽

Vol 14 (3) ◽

pp. 2027-2032

Author(s):

Mita D. Wadekar ◽

J.V. Sathish ◽

C. Pooja ◽

S. Jayashree

Keyword(s):

Treatment Options ◽

Multidrug Resistant ◽

Diffusion Method ◽

Clinical Samples ◽

Beta Lactamase ◽

Beta Lactam ◽

Gram Negative ◽

Extended Spectrum Beta Lactamase ◽

Beta Lactam Antibiotics ◽

Esbl Producers

Resistance to beta lactam antibiotics is the most common cause for beta-lactamase production. Increasing number of extended spectrum beta-lactamase (ESBL) producers has reduced the treatment options which resulted in emergence of multidrug resistant strains, treatment failure and hence increased mortality. To detect phenotypically, ESBL producers in Gram negative isolates from different samples and to know their susceptibility pattern. A retrospective study of Gram negative isolates was conducted. Total of 521 isolates were isolated from various samples. They were processed and identified by standard procedures. The antibiotic susceptibility testing was performed by Kirby- Bauer disc diffusion method using CLSI guidelines. ESBL was detected by combination disk test. A total of 521 Gram negative isolates were isolated which included E. coli, Klebsiella pneumoniae, Citrobacter spp., Enterobacter spp., Proteus spp. and Acinetobacter spp. Pseudomonas aeruginosa. Of 521 isolates tested, ESBL was detected in 329 (63.1%) isolates. These isolates showed maximum susceptibility to piperacillin- tazobactam (86%) followed by imipenem (78.4%), amikacin (63.5%), cotrimoxazole (54.4%), ciprofloxacin (51%), amoxi-clav (44.9%), cefepime (44.1%), gentamicin (38.9%), cefoxitin (34.9%) and ampicillin (19.1%). ESBL producers which are resistant to beta lactam antibiotics have become a major problem. Detection of these beta-lactamase enzymes by simple disk method and its reporting will help clinicians in prescribing proper antibiotics.

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Reconciling the potentially irreconcilable? Genotypic and phenotypic amoxicillin-clavulanate resistance in Escherichia coli

10.1101/511402 ◽

2019 ◽

Cited By ~ 1

Author(s):

Timothy J. Davies ◽

Nicole Stoesser ◽

Anna E Sheppard ◽

Manal Abuoun ◽

Philip Fowler ◽

...

Keyword(s):

Fixed Effects ◽

Beta Lactamase ◽

Beta Lactam ◽

E Coli ◽

Reliable Assessment ◽

Genetic Features ◽

Amoxicillin Clavulanate ◽

Agar Dilution ◽

Phenotypic Methods ◽

Promoter Mutations

AbstractResistance to amoxicillin-clavulanate, a widely used beta-lactam/beta-lactamase inhibitor combination antibiotic, is rising globally, yet susceptibility testing remains challenging. To test whether whole-genome sequencing (WGS) could provide a more reliable assessment of susceptibility than traditional methods, we predicted resistance from WGS for 976 E. coli bloodstream infection isolates from Oxfordshire, UK, comparing against phenotypes from the BD Phoenix (calibrated against EUCAST guidelines). 339/976 (35%) isolates were amoxicillin-clavulanate resistant. Predictions based solely on beta-lactamase presence/absence performed poorly (sensitivity 23% (78/339)) but improved when genetic features associated with penicillinase hyper-production (e.g. promoter mutations, copy number estimates) were considered (sensitivity 82% (277/339); p<0.0001). Most discrepancies occurred in isolates with peri-breakpoint MICs. We investigated two potential causes; the phenotypic reference and the binary resistant/susceptible classification. We performed reference standard, replicated phenotyping in a random stratified subsample of 261/976 (27%) isolates using agar dilution, following both EUCAST and CLSI guidelines, which use different clavulanate concentrations. As well as disagreeing with each other, neither agar dilution phenotype aligned perfectly with genetic features. A random-effects model investigating associations between genetic features and MICs showed that some genetic features had small, variable and additive effects, resulting in variable resistance classification. Using model fixed-effects to predict MICs for the non-agar dilution isolates, predicted MICs were in essential agreement (±1 doubling dilution) with observed (BD Phoenix) MICs for 691/715 (97%) isolates. This suggests amoxicillin-clavulanate resistance in E. coli is quantitative, rather than qualitative, explaining the poorly reproducible binary (resistant/susceptible) phenotypes and suboptimal concordance between different phenotypic methods and with WGS-based predictions.

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Evaluation of the presence of AmpC (FOX) beta-lactamase gene in clinical strains of Escherichia coli isolated from hospitalized patients in Tabriz, Iran

Journal of Experimental and Clinical Medicine ◽

10.52142/omujecm.38.3.17 ◽

2021 ◽

Vol 38 (3) ◽

pp. 301-304

Author(s):

Zahra SADEGHI DEYLAMDEH ◽

Abolfazl JAFARI SALES

Keyword(s):

Escherichia Coli ◽

Antibiotic Resistance ◽

Hospitalized Patients ◽

Disk Diffusion ◽

Beta Lactamase ◽

Beta Lactam ◽

Beta Lactamases ◽

Clinical Strains ◽

Beta Lactam Antibiotics ◽

Lactamase Gene

Beta-lactamases are the most common cause of bacterial resistance to beta-lactam antibiotics. AmpC-type beta-lactamases hydrolyze cephalosporins, penicillins, and cephamycins. Therefore, the study aims was to determine antibiotic resistance and to investigate the presence of AmpC beta-lactamase gene in clinical strains of Escherichia coli isolated from hospitalized patients in Tabriz. In this cross-sectional descriptive study, 289 E. coli specimens were collected from clinical specimens. Disk diffusion method and combined disk method were used to determine the phenotype of extended spectrum β-Lactamase producing (ESBLs) strains. Then PCR was used to evaluate the presence of AmpC (FOX) beta-lactamase gene in the strains confirmed in phenotypic tests. Antibiotic resistance was also determined using disk diffusion by the Kibry-Bauer method. A total of 121 isolates were identified as generators of beta-lactamase genes. 72 (59.5 %) isolates producing ESBL and 49 (40.5 %) isolates were identified as AmpC generators. In the PCR test, 31 isolates contained the FOX gene. The highest resistance was related to the antibiotics amoxicillin (76.12%), ceftazidime (70.24%) and nalidixic acid (65.05%). The results indicate an increase in the prevalence of beta-lactamase genes and increased resistance to beta-lactam antibiotics, which can be the result of improper use of antibiotics and not using antibiotic susceptibility tests before starting treatment. Also, using phenotypic and molecular diagnostic methods such as PCR together can be very useful.

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Intercontinental spread of OXA-48 beta-lactamase-producing Enterobacteriaceae over a 11-year period, 2001 to 2011

Eurosurveillance ◽

10.2807/1560-7917.es2013.18.31.20549 ◽

2013 ◽

Vol 18 (31) ◽

Cited By ~ 166

Author(s):

A Potron ◽

L Poirel ◽

E Rondinaud ◽

P Nordmann

Keyword(s):

Mediterranean Area ◽

Beta Lactamase ◽

Beta Lactam ◽

North African ◽

African Countries ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Beta Lactam Antibiotics ◽

North African Countries ◽

Sequence Types

OXA-48 beta-lactamase producers are emerging as an important threat mostly in the Mediterranean area. We report here the molecular epidemiology of a collection of OXA-48 beta-lactamase-positive enterobacterial isolates (n=107) recovered from European and north-African countries between January 2001 and December 2011. This collection included 67 Klebsiella pneumoniae, 24 Escherichia coli and 10 Enterobacter cloacae. Using the EUCAST breakpoints, ninety-eight isolates (91.6%) were of intermediate susceptibility or resistant to ertapenem, whereas 66% remained susceptible to imipenem. Seventy-five per cent of the isolates co-produced an extended-spectrum beta-lactamase, most frequently CTX-M-15 (77.5%). Susceptibility testing to non-beta-lactam antibiotics showed that colistin, tigecycline, amikacin, and fosfomycin remain active against most of the isolates. Multilocus sequence typing indicated that the most common sequence types (ST) were ST101 and ST38 for K. pneumoniae and E. coli, respectively. The blaOXA-48 gene was located on a 62 kb IncL/M plasmid in 92.5% of the isolates, indicating that a single plasmid was mainly responsible for the spread of that gene. In addition, this study identified multiple cases of importation of OXA-48 beta-lactamase producers at least in Europe, and spread of OXA-48 beta-lactamase producers giving rise to an endemic situation, at least in France.

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First Global Report of Plasmid-Mediated mcr-1 and Extended-Spectrum Beta-Lactamase-Producing Escherichia coli from Sheep in Portugal

Antibiotics ◽

10.3390/antibiotics10111403 ◽

2021 ◽

Vol 10 (11) ◽

pp. 1403

Author(s):

Josman Dantas Palmeira ◽

Marisa Haenni ◽

Jean-Yves Madec ◽

Helena Maria Neto Ferreira

Keyword(s):

One Health ◽

Diffusion Method ◽

Beta Lactamase ◽

Beta Lactam ◽

Colistin Resistance ◽

Disk Diffusion Method ◽

Extended Spectrum Beta Lactamase ◽

E Coli ◽

Beta Lactam Antibiotics ◽

Extended Spectrum

Resistances to extended-spectrum cephalosporins (ESC) and colistin are One Health issues since genes encoding these resistances can be transmitted between all sectors of the One Health concept, i.e., human, animal, and the environment. Among food-producing animals, sheep farming has long been overlooked. To fill in this knowledge gap, we looked for ESC- and colistin resistance in 21 faecal samples collected from sheep in one farm in the south of Portugal. ESC-resistant isolates were selected on MacConkey agar plates supplemented with cefotaxime. Susceptibility testing was performed by the disk-diffusion method according to CLSI, while colistin MIC was determined by broth microdilution. ESC- and colistin-resistance genes were identified by PCR, and the clonality of all isolates was assessed by XbaI-PFGE. The replicon content was determined by PCR according to the PCR-based replicon typing (PBRT) scheme. Sixty-two non-duplicate ESC-resistant E. coli isolates were identified, which all presented an extended-spectrum beta-lactamase (ESBL) phenotype, mostly due to the presence of CTX-M genes. One CTX-M-1-producing E. coli was concomitantly colistin-resistant and presented the plasmid-mediated mcr-1 gene. Nearly all isolates showed associated resistances to non-beta-lactam antibiotics, which could act as co-selectors, even in the absence of beta-lactam use. The results showed a high proportion of ESBL-producing E. coli in sheep faeces. Their dissemination was very dynamic, with the spread of successful clones between animals, but also a large diversity of clones and plasmids, sometimes residing in the same animal. This study highlights the need for global surveillance in all food-producing sectors, in order to avoid the dissemination of genes conferring resistance to last-resort antibiotics in human medicine.

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Molecular characterization and antimicrobial susceptibility profile of New Delhi metallo-beta-lactamase-1-producing Escherichia coli among hospitalized patients

Journal of Laboratory Physicians ◽

10.4103/jlp.jlp_76_17 ◽

2018 ◽

Vol 10 (02) ◽

pp. 149-154

Author(s):

Anjali Agarwal ◽

Jyoti Srivastava ◽

Ujjwal Maheshwari ◽

Mohd Iftikhar

Keyword(s):

Escherichia Coli ◽

Tertiary Care Hospital ◽

Treatment Options ◽

Tertiary Care ◽

Hospitalized Patients ◽

New Delhi ◽

Care Hospital ◽

Beta Lactamase ◽

Cross Sectional ◽

E Coli

ABSTRACT BACKGROUND: Carbapenemase-producing microorganisms are becoming a major concern among hospital-acquired infections. There is also increased multidrug resistance seen among these isolates. AIMS: We have conducted this study to determine the prevalence of New Delhi metallo-beta-lactamase-1 (NDM-1) gene-producing Escherichia coli among hospitalized patients in a tertiary care hospital in Northern India. SETTINGS AND DESIGN: The study was conducted in the Department of Microbiology with the tertiary care hospital settings. It was a prospective cross-sectional observational study conducted during January 2014–August 2014. MATERIALS AND METHODS: A total of 500 nonduplicate E. coli samples were processed. The isolates with reduced susceptibility to ertapenem, i.e., zone diameter between 19 and 21 mm, were considered carbapenemase producers. These isolates were subjected to modified Hodge test for phenotypic confirmation. Polymerase chain reaction was performed on all the screened isolates for molecular detection of NDM-1 gene. STATISTICAL ANALYSIS USED: Chi-square test was used to analyze the data and P < 0.05 was considered statistically significant. RESULTS: Out of 500 E. coli isolates, 61 (12.2%) were screened for carbapenemase production. 47 (9.4%) isolates were positive by modified Hodge test and 36 (7.2%) isolates showed the presence of blaNDM-1 gene (P < 0.05). CONCLUSION: There is an increased prevalence of NDM-1 gene-producing E. coli isolates. These carbapenemase-producing isolates are more resistant to other group of antibiotics (aminoglycosides, fluoroquinolones along with β-lactam group). Early detection of blaNDM-1 gene can help in choosing the effective treatment options for hospitalized patients in time, thereby reducing the risk of mortality.

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