Characterisation of ESBL-producing E. coli isolated from healthy broilers in Tunisia

Foodborne pathogens' transmission is essential in the spread of antibiotic resistance, and extended-spectrum beta-lactamase-producing Escherichia coli especially threatens public health. E. coli plays an essential role in the resistance to commonly used beta-lactam group antibiotics. Ready-to-eat (RTE) stuffed mussels are among many restaurants and street vendors, presenting potential health risks of food hygiene origin. 200 RTE stuffed mussels were collected from the Asian and European sides of Istanbul and analysed for the presence of E. coli. As a result of PCR analysis, E. coli was detected in 7 (3.5%) samples. An antibiotic susceptibility test was performed using the disc diffusion method to determine ESBL and carbapenem resistance. All isolates were resistant to ampicillin. The double-disk synergy test was performed as an ESBL phenotypic confirmation test, and no phenotypically ESBL-producing E. coli were detected. The blaTEM gene was detected in one isolate (14.2%) by mPCR, but blaCTX-M, blaSHV, and blaOXA genes were not observed. Meropenem and imipenem were used with the disk diffusion method for carbapenem resistance study, and no resistant isolate was found. Carbapenem resistance genes were investigated by monoplex PCR, and blaNDM-1, blaOXA-48, blaVIM, and blaIMP resistance genes were not detected. This is the first report on ESBL-producing E. coli in RTE stuffed mussels in Türkiye, which draws attention to a public health risk.

A total of 102 chicken samples were collected from different farms in Qalubia, Behera, Cairo, Assuit and MenofiaGovernorates. The samples were represented by liver, spleen, lungs, heart blood, intestine and kidneys and subjected forisolation and identification of Escherichia coli. The bacteriological examination of the samples indicated the isolation of55 E. coli represented as 31, 6, 8, 5, and 5 from Qalubia, Behera, Cairo, Assuit and Menofia, respectively. Differentserotypes of E. coli (O158, O125, O111, O27, O20, O6, O25, O26, O145, and O159) and12 untyped strains were demonstrated.Thirty-Two E. coli isolates were subjected to initial screening test for extended-spectrum beta-lactamases (ESBLs)production by disc diffusion method with various cephalosporins. The results showed that 46.9% of samples weresensitive to ceftriaxone and cefotaxime and 53.1% to ceftazidime. By double disc synergy test, 19 E. coli strains wereidentified as ESBL producers. PCR results of 32 E. coli isolates showed that blaTEM gene was detected in the genomicDNA of all isolates and in plasmid DNA of 18 isolates. While, blaSHV was detected in the genomic DNA of 12 isolatesand in plasmid DNA of 9 isolates. We can conclude from the current results that amplification of both genomic andplasmid DNA increase the positivity of detection in comparison with amplification of each of DNAs alone

BackgroundThe worldwide prevalence of multi-drug resistance (MDR) in Gram-negative bacteria (GNB), particularly related to extended-spectrum beta-lactamases (ESBLs) and carbapenemases, poses significant global public health and clinical challenges.ObjectivesTo characterize ESBL-producing Gram-negative bacilli, within a pediatric hospital in Gaza using whole genome sequencing (WGS).MethodsA total of 158 clinical isolates of Gram-negative bacilli were collected from Al-Nasser Pediatric Hospital. These isolates were tested for ESBL production using the double disk synergy test. The antibiotic susceptibility profile was determined using the Kirby Bauer method following the Clinical and Laboratory Standard Institute guidelines. Selected 15 phenotypically MDR isolates were whole-genome sequenced and characterized for their genome-based species identity and antibiotic resistance gene profile.ResultsOf the 158 isolates, 93 (58.9%) were positive for ESBL production. The frequency of Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Acinetobacter baumannii, Proteus mirabilis, and Serratia marcescens was 50%, 22.7%, 22.7%, 1.8%, 1.2%, and 1.2% respectively. The prevalence of ESBL among urine, pus, blood, and sputum was 64%, 44%, 23%, and 63.6%, respectively. Chloramphenicol, Imipenem, and Meropenem were the most effective antibiotics against ESBL producers.In sequenced isolates, an average of six anti-microbial resistance (AMR) genes were noted per isolate, where one of them carried up to 13 antibiotic resistance genes. Carbapenem resistance genes such as blaKPC-2(6.6%), blaPDC-36/12 (6.6%), and blaPOM-1 (6.6%) were detected. All the sequenced E. coli isolates (n = 8) showed multiple resistance genes, mainly against β-lactamase (25.0%), aminoglycosides (37.5%), sulfonamides (37.5%), and genes conferring resistance to tetracyclines (25.0).ConclusionOur results showed a high prevalence of ESBL-producing GNB isolated from a pediatric hospital in the Gaza Strip. Various antibiotic resistance genes were identified, including those encoding ESBL and carbapenems. The results highlight the significant challenge posed by MDR in GNB and emphasize the need for effective antibiotic strategies. Given the high endemicity observed in various studies from Palestine, it is important to conduct clinical and molecular epidemiology research to identify risk factors, transmission patterns, and clinical outcomes associated with GNB strains that carry ESBL and carbapenem resistance genes.

A cross sectional study was performed to determine the frequency of Escherichia coli in fresh and frozen meat samples followed by antimicrobial resistance profiling and to detect different extended spectrum beta lactamases (ESBLs) genes. A total of 100 samples of fresh and frozen meat (n=50 each) were collected from different butcher shops and supermarkets. Equal numbers of specimens were collected from chicken and mutton. Samples were processed for isolation and identification of E. coli by standard microbiological, biochemical and molecular characterization. The resistance pattern was detected by Kirby-Bauer disk diffusion method while presence of ESBLs was checked by double disk synergy test and PCR. The results of present study showed that among 100 meat samples, potentially pathogenic E. coli was isolated from 36 samples with greater contamination 20/50 (40%) in chicken samples in comparison to mutton 16/50 (32%). Similarly, the frequency of E. coli was more pronounced in fresh meat 30/50 (60%) rather than frozen 4/50 (8%). The highest resistance pattern (100%) was observed against ampicillin, ciprofloxacin, vancomycin and tetracycline followed by cefotaxime (91.6%) and (n=27) isolates were found multi drug resistant (MDR). The double disk synergy test found 17 (47.22%) ESBL producing isolates while bla CTX-M gene was identified in 5 (29.41%) isolates followed by bla OXA-48 in 4 (23.52%) samples and bla TEM gene in 1 (5.88%). This study revealed that vigilant control procedures should be implemented all over the food chain and effective surveillance should also be performed at national level to minimize the spread of MDR and ESBL producing Escherichia coli from raw meat.

With this study, carbapenem resistance genes were declared for the first time in Enterobacteriaceae isolates isolated from dairy cows’ mastitis infection in Türkiye. In the bacteriological examination of 212 milk samples, 14 (6.60%) E. coli, three (1.41%) Klebsiella oxytoca, and two (0.94%) Klebsiella pneumonia were isolated. At least two E. coli isolates were found to be resistant to all of the antibiotics used in the antibiogram test. The highest resistance was found against cefotaxime and amoxicillin in K. oxytoca isolates. According to the results of PCR targeting blaCTX-M, blaTEM, and blaSHV genes, the blaCTX-M gene was detected in one K. oxytoca and four E. coli isolates, which were found ESBL positive. According to the results of PCR targeting carbapenem and colistin resistance genes, the IMP gene was detected in four E.coli, one K. oxytoca, and one K. pneumonia isolates. OXA-48-like gene was detected in two E. coli isolates. This two E. coli isolates were also IMP gene positive. While NDM gene was detected in two E. coli, KPC gene was detected in one E. coli isolate. One of the colistin resistance genes, mcr-1 was detected in two E.coli strains with PCR. This study showed that ESBL production and carbapenem resistance in Enterobacteriaceae family strains to become prevalent and increasing, especially among E. coli isolates. Furthermore, identification of multiple antibiotic resistance in the isolates indicated that antibiotic resistance also spread rapidly and increased.

Antibacterial resistance patterns of extended spectrum β-lactamase -producing enteropathogenic Escherichia coli strains isolated from children

Event Abstract Back to Event Molecular Characterization of blaTEM and blaCTX-M in ESBLs producing Escherichia coli and Salmonella spp. in Poultry farm and Klebsiella pneumoniae and Proteus spp. in Klang River in Malaysia Nagaraja Suryadevara1*, Kai Boon Yong1, Sharanya Rajan1, Yoke Ing Kwan1 and Michelle Fong Ting Lim1 1 Mahsa University, Malaysia Introduction: Multidrug resistance bacteria (MDR) has emerged as a public health threat nowadays. High level of resistance to beta-lactam antibiotics among Enterobacteriaceae was conferred by Extended Spectrum Beta-Lactamase (ESBL) in the poultry farm. The waste material of the livestock could pollute the river, leading to the aquatic environment as a reservoir of antimicrobial resistance. Methods: The physiochemical properties of water samples were determined. Gram-negative microorganisms were identified using differential media and biochemical testing. Confirmed bacterial isolates were tested for multidrug resistance by using Kirby Bauer method. The ESBL strains were determined phenotypically using Combination Disc Test (CDT) and Double Disc Synergy Test (DDST). Bacterial genomic DNA was extracted from positive ESBL strains, blaTEM and blaCTX-M genes were amplified using Polymerase Chain Reaction (PCR). The amplified products were sequenced to identify the mutated genes in MDR strains. Results: Out of 88 samples collected from poultry farm, 73 (83.0%) E.coli and 59 (67.0%) Salmonella spp. were identified. Of 50 water samples from Klang River, 31 (62.0%) Klebsiella pneumoniae and 13 (26.0%) Proteus spp. were found. In total, 11 (15.1%) E.coli, 14 (23.7%) Salmonella spp., 12 (38.7%) Klebsiella pneumoniae, and 7 (53.8%) Proteus spp. were MDR strains. From there, 8 (72.7%) E.coli, 7 (50.0%) Salmonella spp., 5 (16.1%) Klebsiella pneumoniae and 5 (38.5%) Proteus spp. were ESBL positive strains. The genes blaTEM and blaCTX-M were seen exhibited in 3 (42.9%) and 7 (87.5%) E.coli , 1(14.3%) and 5(71.4%) Salmonella spp., 1 (20.0%) and 4(80.0%) Klebsiella pneumoniae and 1 (20.0%) and 4 (80.0%) Proteus spp., respectively. Conclusion: Classic blaTEM and blaCTX-M genes were present in E.coli and Salmonella spp. from poultry samples; and Klebsiella pneumoniae and Proteus spp. from river water samples. However, blaCTX-M gene was seen more prominent compared to blaTEM gene in the isolated ESBL strains. Acknowledgements Author would like to thank the MAHSA University Management for providing us the necessary laboratory facilities and equipment through out the research. Keywords: MDR - (multi drug resistant), ESBL - Extended-spectrum beta-lactamase, Bla TEM, bla CTX -M1, DDST, CDT Conference: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”, Putrajaya, Malaysia, 3 Dec - 5 Feb, 2019. Presentation Type: Oral Presentation Topic: Miscellaneous Citation: Suryadevara N, Yong K, Rajan S, Kwan Y and Ting Lim M (2019). Molecular Characterization of blaTEM and blaCTX-M in ESBLs producing Escherichia coli and Salmonella spp. in Poultry farm and Klebsiella pneumoniae and Proteus spp. in Klang River in Malaysia. Front. Pharmacol. Conference Abstract: International Conference on Drug Discovery and Translational Medicine 2018 (ICDDTM '18) “Seizing Opportunities and Addressing Challenges of Precision Medicine”. doi: 10.3389/conf.fphar.2018.63.00091 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 22 Oct 2018; Published Online: 17 Jan 2019. * Correspondence: Dr. Nagaraja Suryadevara, Mahsa University, Kuala Lumpur, Malaysia, nagaraja@mahsa.edu.my Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Nagaraja Suryadevara Kai Boon Yong Sharanya Rajan Yoke Ing Kwan Michelle Fong Ting Lim Google Nagaraja Suryadevara Kai Boon Yong Sharanya Rajan Yoke Ing Kwan Michelle Fong Ting Lim Google Scholar Nagaraja Suryadevara Kai Boon Yong Sharanya Rajan Yoke Ing Kwan Michelle Fong Ting Lim PubMed Nagaraja Suryadevara Kai Boon Yong Sharanya Rajan Yoke Ing Kwan Michelle Fong Ting Lim Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. 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Background Extended-spectrum beta-lactamases producing Enterobacteriaceae (ESBL-PE) represent a major problem in wound infections. Here, we investigated the prevalence and molecular characterization of ESBL-PE associated with wound infections in North Lebanon. Research design and methods A total of 103 non-duplicated E. coli and K. pneumoniae strains isolated from 103 patients with wound infections, were collected from seven hospitals in North Lebanon. ESBL-producing isolates were detected using a double-disk synergy test. In addition, multiplex polymerase chain reaction (PCR) was used for the molecular detection of ESBLs genes. Results E. coli was the predominant bacteria (77.6%), followed by K. pneumoniae (22.3%). The overall prevalence of ESBL-PE was 49%, with a significantly higher rate among females and elderly patients. K. pneumoniae was the common MDR and ESBL-producer bacteria (86.95% and 52.17%) compared to E. coli (77.5% and 47.5%). Most of the isolated ESBL producers harbored multiple resistant genes (88%), where blaCTX-M was the most predominant gene (92%), followed by blaTEM (86%), blaSHV (64%), and blaOXA genes (28%). Conclusions This is the first data on the ESBL-PE prevalence associated with wound infections in Lebanon, showing the emergence of multidrug-resistant ESBL-PE, the dominance of multiple gene producers, and the widespread dissemination of blaCTX-M and blaTEM genes.

The increasing prevalence of antimicrobial resistance in livestock, particularly the dissemination of extended-spectrum beta-lactamase-producing Escherichia coli, poses a significant zoonotic and public health risk. This study investigates the genomic characteristics of cefotaxime-resistant E. coli isolates from dairy calves across 23 Czech farms and their caretakers. Bacteriological cultivation on McConkey agar with cefotaxime was used for their isolation, susceptibility to selected antibiotics was determined by disc diffusion method, production of extended-spectrum beta-lactamases (ESBL) was demonstrated by double disc synergy test. The PCR was applied to confirm the presence of selected genes encoding resistance to some beta-lactams and genes encoding resistance to quinolones carried on plasmids. Using whole-genome sequencing, we evaluated resistance genotypes, sequence types, serotypes, plasmid replicons, and virulence genes. Among 266 rectal samples obtained from the calves, 128 (48%) harbored cefotaxime-resistant E. coli. Whole-genome analysis revealed blaCTX-M genes in 91% (116/128) of isolates, with bla CTX-M-14 (44%) and bla CTX-M-1 (34%) being the dominant variants. Other beta-lactamase gene blaTEM-1b was found in 40% (51/128) of isolates. Notably, no cephamycin resistance genes have been identified. The plasmid-mediated quinolone resistance (PMQR) gene qnrS1 was present at 21% (27/128) of isolates. The colistin resistance gene mcr-1 was found in a single ST2325 isolate. Sequence typing revealed significant clonal diversity, with 21 different STs detected among 68 sequenced isolates. ST10 was the most prevalent (27%), followed by ST69 (12%), ST29 (7%) and others. The phylogenetic distribution showed a predominance of commensal groups A (54%) and B1 (21%). The most common serotypes included O101:H9 (21%), O15:H18 (12%), H12, and O70:H11 (7%). Analysis of plasmid content revealed a complex distribution of 18 distinct plasmid replicon types, especially IncF, followed by Col-type and IncI1-type plasmids. Cross-species transmission was indicated by the detection of clonal strains shared between calves and caretakers, notably ST10-O101:H9 and ST34-O68:H30. The prevalence of high-risk clones and the presence of mobile resistance elements underscore the urgent need for stringent monitoring, antimicrobial stewardship, and improved biosecurity measures in livestock environments like increased caution and personal hygiene of animal handlers to mitigate the spread of resistant E. coli between animals and humans.

This study was conducted to determine the presence of ESBL producing Escherichia coli and Klebsiella species in a tertiary care hospital of Bangladesh, as well as to observe the patterns of antibiotic resistance and antibiotic resistance genes among them. A total of 166 Escherichia coli,Klebsiella pneumoniae and Klebsiella oxytoca were isolated from urine, wound swab, pus, sputum and blood samples of patients of Dhaka Medical College Hospital. Antibiotic susceptibility test wasperformed by disk-diffusion technique. ESBL producers were detected phenotypically by Double-disk synergy (DDS) test. Genotypically ESBL genes (blaCTX-M-15, blaOXA-1) among the ESBL producers with presence of class 1 integron among them were detected by PCR. Eighty seven (52.41%) ESBL producers were detected by DDS test. CTX-M-15 (80.46%) was the dominant genotype in ESBL producing strains detected by PCR. Class 1 integron was found in 58 (66.67%) of the phenotypic positive ESBL producers. The results of this study showed high proportion of ESBL producing Escherichia coli and Klebsiella species in Bangladesh.
 Bangladesh J Med Microbiol 2016; 10 (2): 4-8

Background: We investigated the occurrence of blaCTX-M carrying extended spectrum beta lactamase (ESBL) producing Escherichia coli in pigs from 8 North-eastern states of India with special emphasis on the transferability of ESBL gene from resistant E. coli strains to the susceptible Salmonella strains by in vitro and in vivo. Methods: Fecal samples (n=790) were collected from pigs reared under organized and unorganized farming set up of entire North-eastern region of India. All the samples were processed for isolation and identification of E. coli. All the isolates were subjected to antimicrobial sensitivity assay by disc diffusion method followed by determination of ESBLs producing ability by double disc synergy test (DDST). All the ESBLs producing isolates were screened for blaCTX-M gene by PCR using specific primers. The representative blaCTX-M gene positive isolates were used as donor to determine the ability to transfer of resistance gene in Salmonella by in vitro and in vivo assays with and without antibiotic selection pressure. Result: A total of 2,291 E. coli was isolated, of which 1113 and 1178 were from organized and unorganized farms, respectively. Majority of the isolates were multi-drug resistant with highest resistance against amoxicillin (84.81%) followed by cefalexin (77.17%), sulphafurazole (56.79%), piperacillin (46.40%), tetracycline (38.29%) and cefexime (35.66%). Isolates from unorganized farms showed higher resistance than the isolates recovered from organized farms. A total of 654 (28.55%) isolates were confirmed as ESBL producers by double disc synergy test (DDST) method, of which 65 (2.84%) isolates were positive for blaCTX-M gene. Genotypically, isolates with specific amino acids substitution revealed variation in their antibiotic susceptibility by phenotypic method. blaCTX-M gene could be successfully transferred horizontally from E. coli (donor) to Salmonella (recipient) by in vitro (3.6±2.07x10-8 to 4.4±2.88x10-8 transconjugate per donor) and in vivo method. By in vivo method in pig model, the frequency of transfer was higher under the antibiotic selection pressure (6.6±3.05x10-5 to 7.2±1.92x10-5 trans-conjugants per donor) than without antibiotic pressure (5.6±2.3x10-4 to 6.8±3.35x10-4 trans-conjugants per donor).

Background: Rising global concern about antimicrobial resistance (AMR) has drawn attention to the use of antibiotics in livestock. This research was aimed at isolation and determination of the total coliform bacteria, characterization of the bacterial isolates, screening for multidrug resistant bacteria and the resistant genes from faecal samples of pig farms in Anambra State. Methods: A total of 400 pig faecal samples collected from 40 farms in three senatorial zones of Anambra State were subjected to microbiological analysis and the total coliform bacteria determined. Enteric bacteria from the faecal samples were isolated and identified based on their morphological and biochemical characteristics. Susceptibility of the isolates to different antibiotics were carried out using the standard Kirby‐Bauer disc diffusion method and isolates resistant to cephalosporins were further subjected to Double Disc Synergy Test (DDST) for the phenotypical detection of extended spectrum beta-lactamase (ESBL) production. The ESBL producers were subjected to molecular studies for the detection of ESBL genes using PCR protocols. Results: Mean Total coliform counts of the bacteria from pig faecal samples varied in the 3 senatorial zones. Gram-negative bacteria from the pig faecal samples include the genera Escherichia, Klebsiella, Citrobacter, Salmonella, Enterobacter and Proteus. There is a significant difference in resistance of the isolates to cefotaxime (p = 0.025) and streptomycin (p = 0.012) but no significant difference (p > 0.05) was observed on the rest of the antibiotics tested. Streptomycin (60%) was the most highly resisted while Imipenem (4%) was the least antibiotics. 54.6% of all the organisms were multidrug resistant while 43.7% were ESBL producers. BlaCTX was present in ESBL-producing isolates. Conclusion: There is presence of pathogenic enteric organisms in pig faecal samples harbouring antimicrobial resistant genes. Prudent use of antibiotics in pig farms in Anambra State is therefore recommended to reduce the spread of antibiotic resistance.

Antimicrobial resistant Escherichia coli have become an ever increasing problem in human, and animal health and production. The imprudent use of antibiotics and poor hygienic practices especially in poultry industries have been contributing to the emergence and spread of E. coli species resistant to broad spectrum antibiotics including Colistin. This study was conducted to detect colistin - resistance and antibiotic sensitivity patterns in E. coli isolated from broiler chickens in Kelantan. A total of 320 cloacal swabs were collected from apparently healthy broiler chickens in different districts of Kelantan and were analysed using routine microbiological methods, Kirby-Bauer method for antimicrobial susceptibility test and PCR amplification of species-specific and colistin - resistance encoding genes. Out of the 320 samples, 91 isolates were confirmed as E. coli and 21/91 (23.08%) were positive for colistin - resistant encoding gene, mcr-1. Most of the isolates were resistant to tetracycline (95.24%), chloramphenicol (85.71%), and sulphamethoxazole/ trimethoprim (85.71%). However, the isolates were less resistant towards piperacillin/ tazobactam (4.76%) and meropenem (9.52%). The findings from this study reveal the emerging threats of colistin - resistant in local food animal production, particularly in poultry production industry. However, more comprehensive, and large-scale studies focusing on more resistance patterns using determination of minimum inhibitory concentration (MIC), virulence and resistance characteristics and molecular epidemiology of colistin - resistant E. coli are recommended for better understanding of the epidemiology and to implement the appropriate control and prevention strategies.

Background and objectives: Carbapenem resistance is an emerging problem worldwide. Detection of carbapenem resistance genes is important to institute appropriate therapy and to initiate preventive measures. This study was conducted to determine the presence of carbapenemase enzyme producing Escherichia coli and Klebsiella species in a tertiary care hospital of Bangladesh, as well as to observe the patterns of antibiotic resistance and carbapenem resistance genes among them.
 Methodology: Total 166 Escherichia coli, Klebsiella pneumoniae and Klebsiella oxytoca were isolated from urine, wound swab, pus, sputum and blood samples of patients of Dhaka Medical College Hospital. Antibiotic susceptibility test was performed by disk-diffusion technique. Carbapenemase producers were detected phenotypically by Double-disk synergy (DDS) test, Modified Hodge test (MHT) and Combined disk (CD) assay. Minimum inhibitory concentration (MIC) of imipenem was done by agar dilution method among carbapenemase producing strains. Genotypically carbapenemase genes (blaNDM-1, blaVIM, blaIMP, blaKPC, blaOXA-48/blaOXA-181) among the imipenem resistant Escherichia coli and Klebsiella species were detected by polymerase chain reaction (PCR). Sequencing was done to differentiate blaOXA-181 gene from blaOXA-48 gene. Class 1 integron were also detected by PCR using specific primer among carbapenemase producers.
 Results: Thirty seven (22.29%) imipenem resistant isolates were detected during disk-diffusion technique, among them 16 (43.24%) carbapenemase producers were detected by MHT, 20 (54.05%) by DDS test, 22 (59.46%) by CD assay and 23 (62.16%) by PCR. Out of 23 carbapenemase producing strains, MIC of imipenem ranged from 4 μg/ml up to ≥128 μg/ml. NDM-1 (43.24%) was the dominant genotype in imipenem resistant strains followed by KPC (21.62%) and OXA-181 (18.92%). Class 1 integron were present in 16 (69.57%) of the genotypically identified carbapenemase producers.
 Conclusion: The results of this study showed high proportion of carbapenemase enzyme producing Escherichia coli and Klebsiella species in Bangladesh. Genes encoding carbapenemase enzymes including blaNDM-1, blaKPC, blaVIM, blaIMP and blaOXA-181 were responsible for imipenem resistance. blaNDM-1 producers are increasing and blaKPC and blaOXA-181 producers are emerging in Bangladesh. Regular surveillance of antibiotic resistance should be done in every tertiary care hospital to prevent spread of these strains.
 Bangladesh J Med Microbiol 2020; 14 (2): 25-29

Introduction: Antibiotic resistance (ABR) is a serious worldwide threat to public health due to the emergence of multidrug resistant bacteria and is considered as a great problem in the treatment of bacterial infections both in hospital as well as community settings. Extended Spectrum Beta-Lactamase (ESBL) producing strains have emerged as a significant challenge to counter with present antibiotics. Aims: Our study was aimed to know the prevalence of ESBL producing E. coli in our hospital setting for effective therapeutic outcome and patient care. Materials and Method: A total of 459 urine samples of urinary tract infection (UTI) suspects were processed in the Microbiology Department at Jhalawar in Rajasthan, India. All samples were cultured on Mac Conkey Agar and Blood Agar and incubated at 37 °C for 24–48 h. The isolates were identified and confirmed using standard microbiological methods and biochemical reaction. Antibiotic sensitivity testing of all E. coli isolates was performed on Muller Hinton agar plates by Kirby-Bauer disk diffusion technique with guidelines established by the Clinical Laboratory Standards Institute (CLSI). Initial screening of ESBL producing E. coli was performed using the Cefotaxime and Ceftriaxone antimicrobial disc. Double-Disc Synergy Test (DDST) and CLSI confirmatory test i.e., Phenotypic Confirmatory Double-Disc Test (PCDDT) were performed for confirmation of ESBL-producing E. coli. Results: Of the 459 samples processed, 212 isolates were found to be culture positive. Out of 212 positive isolates, 184 (86.79%) were identified as gram negative bacilli (GNB). Of the 184 GNB, 115 (62.50%) were detected as Escherichia coli followed by Klebsiella 30 (16.30%), Pseudomonas 21 (11.41%), Proteus 15 (8.15%), Citrobacter 8 (4.32%), and Acinetobacter 5 (2.71%). Out of 115 E. coli isolates, 86 (74.78%) were found to be ESBLpositive by screening method and 81 (70.43%%) were found to be ESBL producers by PCDDT and 68 (59.13%) were found to be ESBL producers by DDST. Imipenem and Piperacillin–tazobactum were the most active and reliable agents for the treatment of infections which were caused by the ESBL-producing organisms. The ESBL-producing strains were more resistant than non-ESBL producing strains. Among ESBL producers the resistance pattern was highest for Ceftriaxone followed by Cefotaxime and Ceftazidime. Conclusion: As results showed that there was a high prevalence of ESBL production in our setup so, it is essential to report the ESBL production along with the routine sensitivity reports, which will help the clinician in selection of proper antibiotics. Keywords: antibiotic resistance, Extended Spectrum Beta-Lactamase (ESBL), multidrug resistant, Escherichia coli Cite this Article Pawan K, Tiwari YK, Saraf G et al. Identification of ESBL producing Escherichia coli from Urine Samples at Tertiary Care Hospital in Jhalawar. Research & Reviews: A Journal of Microbiology and Virology. 2017; 7(3): 38–45p.