Prevalence Of Extended-spectrum Beta-lactamase Genes Research Articles

Molecular Characteristics of Cephalosporin Resistant Escherichia coli and Klebsiella pneumoniae Isolated from Children in a Tertiary Care Centre of Central Kerala, India

Aims: The study was aimed to determine the molecular characteristics of extended-spectrum beta-lactamases (ESBL) producing cephalosporin-resistant Escherichia coli and Klebsiella pneumoniae isolated from children below ten years of age. Background: Geographically diverse variations in the prevalence of ESBL genes were reported. No data were available on the prevalence of ESBL genes in central Kerala, India, among children below 10 years of age. Methods: A cross-sectional study was performed to analyze ESBL genes in cephalosporin-re-sistant E. coli and K. pneumoniae strains isolated from samples received in the Microbiology la-boratory of a tertiary care centre during the period between May 2021 and July 2022. The strains showed that ESBL + cephalosporin resistance was subjected to PCR-based genotyping for the genes such as bla (beta-lactamase) CTX-M-1, blaCTX-M-15, blaCTX-M-U, blaTEM, blaPER and SHV. Results: Among the total 228 samples analyzed, 136 (60%) had no growth. Ninety-two (40 %) samples showed growth of E. coli and K. pneumoniae. Among the isolates that showed growth, 39 (42%) were sensitive, and the remaining 53 (57%) were resistant to third-generation cephalospor-ins. Among the isolates showed resistance, 22 (42%) were ESBL positive and 31 (58%) were ESBL negative. Among the positive ESBL, nine E. coli strains (60%) were positive for CTX-M-15 and CTX-M-1. CTX-M-15 and CTX-M-U were present in six (85%) K. pneumoniae with ESBL +. Conclusion: E. coli and K. pneumoniae isolated from specimens of children below ten years of age showed 41-42% ESBL producers. Prevalent ESBL-producing genes in E. coli were CTX-M-15 and CTX-M-1. CTX-M-15 and CTX-M-U were prevalent in ESBL-producing K. pneumoniae.

Prevalence of blaCTX-M and blaTEM Genes in Cefotaxime-Resistant Escherichia coli Recovered from Tertiary Care at Central Nepal: A Descriptive Cross-Sectional Study.

Urinary tract infections (UTIs) are highly prevalent globally, and various antibiotics are employed for their treatment. However, the emergence of drug-resistant uropathogens towards these antibiotics causes a high rate of morbidity and mortality. This study was conducted at the Microbiology Laboratory of Grande International Hospital from November 2021 to May 2022 and aimed to assess the prevalence of UTI caused by Escherichia coli and their antibiotic susceptibility pattern with a focus on extended-spectrum beta-lactamases (ESBLs) and the prevalence of two genes (blaCTX-M and blaTEM) in cephalosporin-resistant E. coli. Altogether, 1050 urine samples were processed to obtain 165 isolates of E. coli. The isolates were identified by colony morphology and biochemical characteristics. Antimicrobial susceptibility tests (ASTs) were determined by the Kirby-Bauer disk diffusion method, and their ESBL enzymes were estimated by the combined disk method (CDM). Two ESBL genes (blaCTX-M and blaTEM) were investigated by polymerase chain reaction (PCR) in cefotaxime-resistant E. coli. Among the 1050 urine samples that were processed, 335 (31.9%) were culture-positive with 165 (49.2%) identified as E. coli. The age group ≥60 years (30.3%) had greater susceptibility to bacterial infections. AST revealed that meropenem was highly effective (95.7% susceptibility), while ampicillin showed the least sensitivity (42.4%). Among the E. coli isolates, 86 were multidrug resistant (MDR) and 10 were extensively drug resistant (XDR). Of these, 46 MDR (96%) and 2 XDR (4%) were ESBL producers. The prevalence of ESBL genes (blaCTX-M and blaTEM) was 49.3% and 54.8%, respectively. The overall accuracy of CDM as compared to PCR for the detection of the blaCTX-M gene was 55.26%. The prevalence of MDR E. coli harboring the blaCTX-M and blaTEM genes underscores the imperative role of ESBL testing in accurately identifying both beta-lactamase producers and nonproducers.

Prevalence of ESBL genes in Klebsiella pneumoniae from individuals with community-acquired urinary tract infection in rural communities of Enugu State, Nigeria

Background: The emergence and spread of extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae is now a major challenge in treating urinary tract infections. This study was conducted to determine the Prevalence of ESBL genes in Klebsiella pneumoniae and the influence of these genes on the antimicrobial susceptibility patterns of the isolates, obtained from community acquired urinary tract infection in Enugu state rural communities. Method: A total of 735 clean catch mid-stream urine samples were collected from February, 2021 to June, 2021. The urine samples were cultured and Klebsiella pneumoniae were identified morphologically, biochemically, and then typed down, using PCR, Gel electrophoresis, and Sanger Sequencing by Genewiz. Antibiotic susceptibility testing was done by modified Kirby-Bauer disc diffusion method and interpretation was done following Clinical and Laboratory Standard Institute (CLSI) guidelines. ESBL screening was done by the phenotypic method, using the standard disk diffusion method, whereas the phenotypic confirmation of ESBL producers was done by the double-disk synergy method, and interpreted according to CLSI guidelines. Multiplex PCR was used to detect the genes for SHV and CTX-M while conventional linear PCR was done for TEM and blas GES type ESBL genes. Results: A total of 77 isolates were identified as Klebsiella pneumoniae, and the prevalence of positive ESBL in them were 10(12.9%). Of the 77 isolates that were tested for antibiogram, 29 isolates were multidrug-resistance (MDR). Out of the MDR isolates, 10 were ESBL positive, whereas 19 were not. Of this 29 MDR isolates, 6 were extensively-drug resistant (XDR). Of the 6 XDR isolates, 3 do not possess ESBL enzymes, whereas 3 of them do. Age range of 31-40 contributed the highest prevalence of ESBL genes (60%), whereas age range of 10-20, 21-30, and 51-70 did not produce any ESBL gene. bla TEM gene was the most prevalent ESBL resistant genes with a prevalence of 10 (100%), followed by SHV 9(90%), whereas bla GES gene and bla CTX-M have the least prevalence of 4 (40%) each. Conclusion: Higher prevalence of MDR, XDR, and ESBL-producing Klebsiella pneumoniae were observed, thus the need for public health intervention for effective prevention and control of antimicrobial resistance, and proper treatment of UTIs.

Antimicrobial Resistant Bacteria in Health Care Facilities: Exploring Links With Water, Sanitation, and Hygiene in Gaza, Palestine

Background Antimicrobial resistance (AMR) is a growing global phenomenon; however, its link to water, sanitation, and hygiene (WASH) remains underexplored, particularly in health care facilities where humanitarian crises prevail. Objective This study aimed to identify AMR bacteria in samples collected from WASH services in 2 hospitals in Gaza and to investigate the presence of AMR genes. Methods A hospital-based cross-sectional study to detect and identify antimicrobial resistance bacteria was conducted. Random samples from water, wastewater, soap, and surface swabs (n=345) were collected from Al-Shifa and European Gaza hospitals and screened for the presence of Enterobacteriaceae, Pseudomonas, Enterococcus, and Staphylococcusaureus. Antimicrobial susceptibility, extended spectrum beta-lactamase (ESBL) production, carbapenem resistance, and AMR genes were investigated. Results High levels of bacterial contamination were detected in water and surface swab samples with an overall percentage of 34.1%. Moreover, 22% of the identified Enterobacteriaceae was positive for ESBL, and 14% was positive for modified Hodge test. Over 2/3 of isolated Enterobacteriaceae in water and wastewater samples was found to be resistant to amikacin, ceftazidime, ceftriaxone, and imipenem. All Enterobacteriaceae isolates from swab samples were found to be resistant to piperacillin-tazobactam, amikacin, ceftazidime, and ceftriaxone; 13.8% of S. aureus in water samples was methicillin resistant. The prevalence of ESBL genes among Enterobacteriaceae isolates was 25% OXA, 19.4% SHV, 2.8% KPC, 66.7% TEM, 41.7% blaCTXM, and 5.6% blaCTXM-3. For carbapenem-resistant gene (MDM), the prevalence among Enterobacteriaceae was 11.1%, and among Pseudomonas was 12.5%. The antibiotic susceptibility profile was also presented for Pseudomonas, Enterococcus, and S. aureus. Conclusions The results underline the level of contamination with AMR bacteria in WASH samples and highlight the need to consider the safety of WASH service at health care facilities as an essential aspect in the fight against the spread of AMR and to interrupt nosocomial transmission.

Erratum to: Antimicrobial resistance and prevalence of extended-spectrum beta-lactamase genes in Escherichia coli from major rivers in Podhale, southern Poland

The text in the original article on page 246 left hand column contains errors.

Antimicrobial resistance and prevalence of extended-spectrum beta-lactamase genes in Escherichia coli from major rivers in Podhale, southern Poland

The aim of this study was to assess the antimicrobial resistance and the prevalence of genes determining the presence of extended-spectrum beta-lactamase (ESBL) enzymes in Escherichia coli isolated from two major rivers of the Podhale region in southern Poland. In total, 196 E. coli isolates were analyzed—98 from each river—Białka and Zakopianka, collected in 8 campaigns, over the period of two years. Antimicrobial resistance was assessed using disk diffusion method and PCR tests were conducted to detect the ESBL genes. In E. coli isolated from Białka, the resistance to amoxicillin/clavulanic acid was detected most frequently (54.08%) and ESBL was detected in 14.29% of strains. In strains isolated from Zakopianka, most frequent resistance was observed toward ticarcillin (51.02%), while ESBL was observed in 16.33% of isolates. In the total pool of isolates, the resistance to amoxicillin/clavulanic acid was most frequent (48.98% of isolates) and ESBL producers comprised 15.30% of E. coli isolates derived from both rivers. Multidrug resistance was observed less frequently in strains derived from Białka (4 isolates resistant to 10 and more antimicrobials) than from Zakopianka, where 10 isolates were resistant to 10 and more antibiotics. Out of the tested ESBL genes blaTEM was detected most frequently (45.4% of isolates), whereas blaCTX-M1 and blaCTX-M3 were recorded in one isolate.

National sentinel site surveillance for antimicrobial resistance in Klebsiella pneumoniae isolates in South Africa, 2010 - 2012.

The increasing rates of antimicrobial resistance observed in the nosocomial pathogen Klebsiella pneumoniae are of major public health concern worldwide. To describe the antibiotic susceptibility profiles of K. pneumoniae isolates from bacteraemic patients submitted by sentinel laboratories in five regions of South Africa from mid-2010 to mid-2012. Molecular methods were used to detect the most commonly found extended-spectrum beta-lactamase (ESBL) and carbapenemase resistance genes. Thirteen academic centres serving the public healthcare sector in Gauteng, KwaZulu-Natal, Free State, Limpopo and Western Cape provinces submitted K. pneumoniae isolates from patients with bloodstream infections. Vitek 2 and MicroScan instruments were used for organism identification and susceptibility testing. Multiplex polymerase chain reactions (PCRs) were used to detect blaCTX-M, blaSHV and blaTEM genes in a proportion of the ESBL isolates. All isolates exhibiting reduced susceptibility to carbapenems were PCR tested for blaKPC and blaNDM-1 resistance genes. Overall, 68.3% of the 2,774 isolates were ESBL-positive, showing resistance to cefotaxime, ceftazidime and cefepime. Furthermore, 46.5% of all isolates were resistant to ciprofloxacin and 33.1% to piperacillin-tazobactam. The major ESBL genes were abundantly present in the sample analysed. Most isolates (95.5%) were susceptible to the carbapenems tested, and no isolates were positive for blaKPC or blaNDM-1. There was a trend towards a decrease in susceptibility to most antibiotics. The high proportion of ESBL-producing K. pneumoniae isolates observed, and the prevalence of ESBL genes, are of great concern. Our findings represent a baseline for further surveillance in SA, and can be used for policy and treatment decisions.

PER, CTX-M, TEM and SHV Beta-lactamases in Clinical Isolates of Klebsiella pneumoniae Isolated from Tehran, Iran

Objective(s) Different types of extended spectrum beta-lactamases (ESBLs) are encountered in the clinical settings worldwide. There are a few studies regarding the prevalence of ESBL genes among Klebsiella pneumoniae isolates at Tehran especially those of blaPER and blaCTX. The aim of this study was to determine the prevalence of blaSHV, blaTEM ,blaPER and blaCTX genes among clinical K. pneumoniae of different hospitals in Tehran. Materials and Methods Two hundred isolates of K. pneumoniae were received from different clinical specimens. The susceptibility of the isolates to 10 different antibiotics was examined by disk diffusion test. The MICs for ceftazidime were also determined using micro-broth dilution assay. Isolates showing MIC≥ 4 µg/ml for ceftazidime were screened for ESBL production by phenotypic confirmatory test (PCT) and subjected to PCR for studied genes. Variation among four amplified genes was evaluated using PCR-RFLP. Results By disk diffusion test, resistance to ceftazidime and cefotaxime were 34.7% and 33.5% respectively. However, all strains were susceptible to imipenem. Eighty isolates showed MICs≥ 4 µg/ml for ceftazidime of which 77 (96%) were positive for ESBL in PCT. The prevalence of blaSHV, blaCTX-M, blaTEM and blaPER among these isolates were 26%, 24.5%, 18% and 7.5%, respectively. No variation was detected in the genes by PCR-RFLP. Conclusion As far as we know this is the first report of the blaPER-1 in K. pneumoniae in Iran. The blaCTX-M was the second most common gene detected among the ESBL positive isolates of K. pneumoniae. For rapid identification of ESBL producing isolates it was recommended that clinical laboratories adopt simple test based on CLSI recommendation for confirming ESBL production in enterobacterial species.

Prevalence and molecular characterization of extended-spectrum β-lactamase-producing Klebsiella pneumoniae in Riyadh, Saudi Arabia

BACKGROUND AND OBJECTIVES:Reports on extended-spectrum β-lactamases (ESBL) production by Enterobacteriaceae, and especially in Klebsiella pneumoniae, are few in Saudi Arabia. Therefore, we determined the prevalence of ESBL in K pneumoniae from Riyadh and characterized the predominant β-lactamase gene in these isolates.METHODS:A total of 400 K pneumoniae samples were isolated from two hospitals in Riyadh during 2007 and screened for production of ESBL using ESBL-E-strips and combined disk methods. PCR assay was used to detect blaTEM, blaSHV, and blaCTX-M genes.RESULTS:Phenotypic characterization identified a high ESBL rate of 55% of K pneumoniae isolates. ESBL-producing K pneumoniae were PCR positive for SHV, TEM and CTX-M β-lactamase genes with prevalences 97.3%, 84.1% and 34.1%, respectively. Within the CTX-M family, two groups of enzymes, CTX-M-1 and CTX-M-9-like genes were found with prevalences of 60% and 40%, respectively.CONCLUSIONS:This study confirms the high rate of ESBL in K pneumoniae clinical isolates in hospitals in Riyadh. This study demonstrates the worldwide spread of blaCTX-M genes. This first report of the presence of the blaCTX-M gene in clincial isolates in Saudi Arabia is evidence of the continuing worldwide spread of this gene.