Detection Of Extended-spectrum Beta-lactamases Research Articles

A Hemagglutination, serum resistance, cell surface hydrophobicity and ESBL production among uropathogenic Escherichia coli

Abstract
 Background
 Bacteria have adapted themselves with increased pathogenesis and drug resistance mechanisms to overcome host defense systems. Uropathogenic E. coli (UPEC) responsible for symptomatic Urinary tract infections (UTIs) carry diverse virulence factors and extended spectrum beta-lactamase (ESBL) production which play pivotal role in pathogenesis and antibiotic resistance. Objective: The purpose of this study was to determine the phenotypic expression of virulence factors; i.e. Hemagglutination, serum resistance, cell surface hydrophobicity and resistance factor ESBL production among UPEC isolates. Methods: In this prospective study 50 drug resistant UPEC strains were evaluated for hemagglutination by slide method, cell surface hydrophobicity by salt aggregation test, serum resistance by serum bactericidal assay and ESBL detection by double disc synergy test. Results: Among 50 UPEC strains 46 (92%) showed mannose resistant hemagglutination, 48 (96%) were found to be serum resistant, 40 (80%) isolates were positive for cell surface hydrophobicity, while 43 (86%) were ESBL producers. Most of the UPEC that expressed virulence factors were ESBL producers as well. Conclusion: Over all, this study indicated that MDR UPEC isolates have expressed a number of virulence factors with ESBL production, which explains how pathogens have increased their virulence and resistance repertoire resulting in treatment failures.

Detection of Extended-Spectrum Beta-Lactamase (ESBL)-Producing Enterobacteriaceae from Diseased Broiler Chickens in Lusaka District, Zambia.

Poultry products in Zambia form an integral part of the human diet in many households, as they are cheap and easy to produce. The burden of poultry diseases has, however, remained a major challenge. Growing consumer demand for poultry products in Zambia has resulted in non-prudent antimicrobial use on farms, intending to prevent and treat poultry diseases for growth optimisation and maximising profits. This cross-sectional study aimed to identify the different types of bacteria causing diseases in chickens in Lusaka and to detect the extended-spectrum lactamase (ESBL)-encoding genes. We collected 215 samples from 91 diseased chickens at three post-mortem facilities and screened them for Gram-negative bacteria. Of these samples, 103 tested positive for various clinically relevant Enterobacteriaceae, including Enterobacter (43/103, 41.7%), Escherichia coli (20/103, 19.4%), Salmonella (10/103, 9.7%), and Shigella (8/103, 7.8%). Other isolated bacteria included Yersinia, Morganella, Proteus, and Klebsiella, which accounted for 21.4%. E. coli, Enterobacter, Salmonella, and Shigella were subjected to antimicrobial susceptibility testing. The results revealed that E. coli, Enterobacter, and Shigella were highly resistant to tetracycline, ampicillin, amoxicillin, and trimethoprim-sulfamethoxazole, while Salmonella showed complete susceptibility to all tested antibiotics. The observed resistance patterns correlated with antimicrobial usage estimated from sales data from a large-scale wholesale and retail company. Six (6/14, 42.9%) E. coli isolates tested positive for blaCTX-M, whilst eight (8/14, 57.1%) Enterobacter samples tested positive for blaTEM. Interestingly, four (4/6, 66.7%) of the E. coli isolates carrying blaCTX-M-positive strains were also positive for blaTEM. Sanger sequencing of the PCR products revealed that five (5/6, 83.3%) of the abovementioned isolates possessed the blaCTX-M-15 allele. The results suggest the presence of potentially pathogenic ESBL-producing Enterobacteriaceae in poultry, threatening public health.

Prevalence of extended-spectrum beta-lactamase and molecular detection of blaTEM, blaSHV, and blaCTX-M genotypes among gram-negative Bacilli isolates from hospital acquired infections in pediatrics, one institutional study

BackgroundGram-negative bacilli represents an important pathogen in hospital-acquired infections (HAIs) worldwide. The emergence of antibiotic resistance in these pathogens warrants attention for the proper management of infections. Extended-spectrum beta-lactamase (ESBL) resistance represents a major therapeutic problem in infections due to Gram-negative bacilli.The present study aimed to study the extended-spectrum beta-lactamase genes blaTEM, blaSHV, and blaCTX-M by multiplex polymerase reaction in isolated Gram-negative bacilli from HAIs in pediatric patients.MethodsThe study included one hundred-five isolates of Gram-negative bacilli from pediatric patients with different types of HAIs. The isolates were subjected to full microbiological identification, antibiotics susceptibility by disc diffusion method, the phenotypic study of ESBL, and the genetic study of ESBL genes by multiplex PCR.ResultsFifty isolates of Gram-Negative bacilli showed ESBL activity by a phenotypic study by double disc diffusion method (50/105). All ESBL producers’ isolates were positive by PCR for ESBL genes. The most frequent gene was blaTEM (64%), followed by blaSHV (30%) and CTX-M (22%). Mixed genes were found in 4 isolates (8%) for blaTEM and blaSHV, blaTEM and CTX-M. There was a significant association between PCR for ESBL genes and phenotypic ESBL detection (P = 0.001). There was significant detection of ESBL genes in E. coli (28%), followed by Enterobacter spp. (26%), Klebsiella spp. (24%), Serratia (14%), Pseudomonas spp. (6%) and Proteus (2%), P = 0.01. There Seventy percent of isolates positive for ESBL production had an insignificant association between MDR and PCR for ESBL genes (P = 0.23).ConclusionThe present study highlights the prevalence of ESBL activity among clinical isolates of Gram-negative bacilli isolated from hospital-acquired infections in pediatric patients. The most common gene responsible for this activity was blaTEM gee followed by blaSHV and blaCTX-M. There was a high prevalence of multiple antibiotic resistance among isolates with ESBL activity. The finding of the present study denotes the importance of screening extended beta-lactamase among Gram-negative bacilli associated with HAIs in pediatric patients.

Prevalence of drug resistant Enterobacteriaceae in a Nepalese tertiary care hospital.

Antimicrobial resistance in Enterobacteriaceae is an emerging global public health problem. Numerous studies have reported community-acquired AmpC beta-lactamase and extended spectrum beta-lactamase (ESBL) producing Enterobacteriaceae in Nepal. However, there are limited data on community-acquired Metallo-beta-lactamase (MBL) producing Enterobacteriaceae. A hospital-based descriptive cross-sectional study was conducted using 294 Enterobacteriaceae isolates from a total of 2,345 different clinical specimens collected from patients attending a tertiary care hospital in Nepal. Bacteria were isolated using standard microbiological growth media and identified using biochemical tests. For antimicrobial susceptibility testing, Kirby-Bauer disc diffusion technique was used. AmpC, ESBL, and MBL productions were detected by using combined disc method. AmpC, ESBL, and MBL productions were detected in 19.4%, 29.6%, and 8.5% of total Enterobacteriaceae isolates respectively. Higher rates of beta-lactamases production were seen among the isolates from in-patients in comparison with those from out-patients. However, 11.6%, 25%, and 3.7% of the total isolates from out-patients were AmpC, ESBL, and MBL producers respectively. The co-production of the beta-lactamases was also detected, with two Klebsiella pneumoniae isolates producing all three beta-lactamases. One MBL producing Proteus vulgaris isolate that was pan-resistant with no remaining treatment options was also isolated. Prevalence of drug resistant Enterobacteriaceae in our study was very high. Detection of AmpC, ESBL, and MBL positive isolates from out-patients, who did not have recent history of hospital visit, indicated the community dissemination of the drug resistant bacteria. This is a matter of great concern and an immediate attention to formulate strategies to prevent further development and spread of antibiotic resistance is required.

Detection of Antimicrobial Susceptibility Pattern and Molecular Detection of Resistance Genes in Klebsiella pneumoniae Isolated from Urine Samples of Suspected UTI Patients Attending Public Hospitals in Nasarawa South Senatorial District, Nasarawa State, Nigeria

Aims: The aim of the current study was to determine antimicrobial susceptibility patterns and the incidence of resistance genes (β-lactamase-encoded genes) among clinical isolates of K. pneumoniae.
 Study Design: Investigative.
 Place and Duration of Study: Department of Microbiology, Nasarawa State University, Keffi, between November 2022 and July 2023.
 Methodology: A total of 375 clinical samples were randomly collected from patients and isolation and identification was done using standard microbiological methods. The antibiotic susceptibility testing for the isolates was carried out in accordance with the Clinical and Laboratory Standards Institute protocol. The detection of extended-spectrum beta-lactamase (ESBL) production in K. pneumoniae isolates was carried out using double disc synergy test. In addition, molecular detection of ESBL genes was carried out using Polymerase Chain Reaction (PCR) method.
 Results: The overall incidence rate of K. pneumoniae in the 375 clinical samples from Nasarawa South Senatorial District was 38 (10.13%). Resistance was high to ampicillin (94.74%) and amoxicillin-clavulanic acid (73.6%) but low to cefotaxime (13.16%), ceftriaxone (18.42%), ceftazidime and cefoxitin (23.60%) and gentamicin (28.94%). Most of the isolates were multi-drug resistant (MDR; 81.5%) and none was Pan Drug resistant. All 15 isolates jointly resistant to cefotaxime and/or ceftazidime and ciprofloxacin 100.0% were ESBL producers, distributed as follows: blaCTX-M-1 7(46.6%); blaCTX-M-34(26.6%); blaSHV 6(40.0%); blaTEM6 (40.0%); and blaOXA 5(33.3%).
 Conclusion: K. pneumoniae isolated from the urine of patients attending public hospitals in Nasarawa South were less resistant to cefotaxime, ceftazidime, gentamicin, ceftriaxone, and cefoxitin in the study location. This implies that the antibiotics are useful in the treatment of infection caused by K. pneumoniae. Also, ESBL-positive K. pneumoniae isolates harbored ESBL genes, with blaCTX-M as the most common.

A-233 Novel Microfluidic System for Antimicrobial Susceptibility Testing

Abstract Background Aiming to enhance antibiotic susceptibility testing (AST), BacterOMIC combines microdilution AST with cutting-edge microfluidics to quantitatively evaluate all clinically relevant antibiotics on a single disposable panel. In this study, we evaluated the performance of BacterOMIC in determining minimum inhibitory concentrations (MICs) of selected agents against Gram-positive and Gram-negative bacteria and in detecting the presence of extended spectrum beta-lactamase (ESBL) produced by Enterobacterales. Methods Using the BacterOMIC UNI Panel, we characterized bacterial strains collected from Polish hospitals in 2020–2022. We inoculated the BacterOMIC panels with bacterial suspensions in cation-adjusted Mueller-Hinton broth at a density of 5*10^5 CFU/mL. The panels then incubated in an analyzer for 16 h, and results were automatically calculated via built-in analytical software. For comparison, we obtained reference MIC data from laboratories contracted for determining strain MIC values via microdilution. We calculated the essential agreement (EA%) and bias according to ISO20776.We also qualitatively assessed several antibiotics via our panel. Categorical agreement (CA%) and discrepancies were determined for these antibiotics according to ISO20776.To assess ESBL detection, we compared BacterOMIC results with data obtained in our internal laboratory using the MIC Test Strip ESBL (Liofilchem). Results With the BacterOMIC system, CA% values exceeded 90% for amoxicillin, benzylpenicillin, ceftazidime/avibactam, ciprofloxacin, erythromycin, gentamicin, meropenem, moxifloxacin, tedizolid, and vancomycin. EA% values exceeded 90% for the following antimicrobials, with a bias of less than 30%. By comparing with our reference method, we found that BacterOMIC detected ESBL with a specificity of 92% (confidence interval [CI] 95%; 81%–98%) and sensitivity of 97% (CI 95%; 84%–99%). Conclusions Our data demonstrate that BacterOMIC provides reliable MIC and ESBL results for assessing the drug susceptibility of Gram-positive and Gram-negative bacteria against 25 agents in a single panel. Thus, the BacterOMIC system provides a valuable tool for automated AST systems.

Detection of Extended Spectrum Beta Lactamase Producing Escherichia Coli and Klebsiella Species Isolated from Urine Samples of UTI Patients by Phenotypic and Genotypic Methods

Background: Escherichia coli and Klebsiella spp. are the most common organisms causing urinary tract infection (UTI) and commonly responsible for extended spectrum beta lactamase (ESBL) production. This study was carried out to detect ESBL producing uropathogenic Escherichia coli and Klebsiella spp. by phenotypic and genotypic method. Rapid and accurate detection of ESBL producing E. coli and Klebsiella spp. has an important role to avoid treatment failure. Materials and Methods: This was a cross sectional observational study and carried out in department of microbiology in Chittagong Medical College, Bangladesh from January to December 2017. Urine was collected from suspected UTI patients and standard microbiological and biochemical tests were carried out. ESBL producing E. coli and Klebsiella spp. were identified by phenotypic confirmatory disc diffusion test (PCDDT). Polymerase chain reaction (PCR) was performed by using standard protocol with specific primers. Results: 448 urine samples were collected. Among them, 140 showed bacterial growth; 72 were E. coli and 35 were Klebsiella spp. Among E. coli 38(52.8%) and in Klebsiella spp. 15(42.9%) were detected as ESBL producers by PCDDT respectively. Among E. coli, 41(56.9%) and in Klebsiella spp. 21(60%) strains produced ESBL genes by PCR. Out of 38 phenotypically positive E. coli, 7 strains do not carry any detectable genes. Similarly, out of 15 phenotypically positive Klebliella spp. 3 isolates did not produce any detectable gene. On the other hand, 10 E. coli isolates and 9 Klebsiella spp. carry detectable genes although these were not phenotypically ESBL producers. Moreover, ESBL producing E. coli and Klebsiella spp. showed more multidrug resistant than Non-ESBL producing E. coli and Klebsiella spp. Conclusion: This study revealed that large portion of E. coli and Klebsiella spp. was ESBL producers. PCR can detect some additional cases of ESBL producing isolates. So, PCR can be used along with phenotypic ......

Genotypic characterization of extended spectrum beta-lactamase in gram negative bacterial contaminants of some door handles in Olabisi Onabanjo University Teaching Hospital, Sagamu, Ogun state.

Nosocomial infections due to cross-transmission by microorganisms from inanimate surfaces have become a recognized public health problem. This research was carried out to detect bacterial contaminants of door handles and their production of extended spectrum beta-lactamase (ESBL) in Olabisi Onabanjo University Teaching Hospital. Seventy door handles from twenty-seven sections of Olabisi Onabanjo University Teaching Hospital were swabbed and inoculated on different culture media. The isolates were identified using standard microbiological procedures. Susceptibility of the isolates to eleven antibiotics was determined by Kirby- Bauer disc diffusion test. Phenotypic detection of ESBL was determined by modified double disc synergy test (MDDST) followed by PCR amplification of ESBL- encoding genes. Forty-four (62.9%) out of 70 door swabs were positive for bacterial growth. S . aureus was the most common isolate (22.9%). Multidrug resistance was observed in 15(79.0%) and 20(69%) of Gram positive and Gram-negative isolates, respectively. Coagulase negative staphylococci, E. coli, Acinetobacter and Shigella species were totally resistant to ceftazidime (100%). PCR amplification of ESBL-genes revealed the presence of blaTEM blaCTX -M and blaSHV in 4(14.3%), 2(7.2%) and 1(3.6%) isolate, respectively. This study revealed that door handles of different sections in hospital environment were contaminated by multidrug resistant Gram-negative bacteria encoding ESBL genes.

EXTENDED SPECTRUM BETA LACTAMASE PRODUCING UROPATHOGENIC ESCHERICHIA COLI AND THEIR ANTIMICROBIAL SUSCEPTIBILITY PATTERN IN A TERTIARY CARE HOSPITAL

Urinary tract infection (UTI) due to extended spectrum beta-lactamase (ESBL)-producing bacteria including Escherichia coli has become widespread and also there is a changing trend in the antibiotic susceptibility pattern to the conventional drugs used in the treatment of urinary tract infections due to the production of extended-spectrum beta-lactamases (ESBLs).This study was done to nd the percentage of ESBL producing Escherichia coli and to study the antibiotic resistance prole of the ESBL and non-ESBL producing Escherichia coli. Urine samples were processed and identication of bacterial growth was conrmed by standard microbiological procedures. Antimicrobial susceptibility and ESBL detection were performed using Clinical and Laboratory Standards Institute (CLSI M100 2022) guidelines, ESBL detection was done by combined disc method using cefotaxime (30 ug) versus cefotaxime plus clavulanate (30+10 ug). Out of total 1238 urine samples 549 were found to be culture positive. Escherichia coli (36.2%) was the most frequently isolated uropathogen followed by Klebsiella pneumoniae (23.6%). Among the isolated Escherichia coli 44.2% were ESBL producers which were found highly resistant in comparison to non-ESBL producers.

Nosocomial infections and antimicrobial susceptibility patterns among patients admitted to intensive care unitofImam Khomeinihospitalin Ilam, Iran.

Nosocomial infections (NIs) are a major challenge worldwide. Identification of antibiotic resistance pattern extended spectrum beta-lactamases (ESBLs) and carbapenem-resistant Enterobacteriaceae (CRE) were the objectives of this study. In this cross-sectional study, the antimicrobial susceptibility pattern of bacterial isolates collected from patients with NIs in ICU was determined. Overall, 42 Escherichia coli and Klebsiella pneumoniae isolates from different infection sites were used to determine phenotypic tests of ESBLs, Metallo-β-lactamases (MBLs) and CRE. Detection of ESBLs, MBLs and CRE genes were performed by the polymerase chain reaction (PCR) method. From 71 patients with NIs, 103 different bacterial strains were isolated. The most frequently isolated bacteria were E. coli (n = 29; 28.16%), Acinetobacter baumannii (n = 15; 14.56%), and K. pneumoniae (n = 13; 12.26%). Also, the rate of multidrug-resistant (MDR) isolates was 58.25% (60/103). Based on phenotypic confirmation tests, 32 (76.19%) isolates of E. coli and K. pneumoniae produced ESBLs, and 6 (14.28%) isolates were identified as CRE producers. PCR showed the high prevalence of the blaCTX-M (n = 29; 90.62%) in ESBL genes. In addition, blaNDM was detected in 4 (66.66%), blaOXA-23 in 3 (50%), and blaOXA-48 gene in 1 (16.66%) isolates. The blaVIM, blaKPC, and blaIMP genes were not detected in any of the isolates. The Gram-negative bacteria E. coli, A. baumannii, and K. pneumoniae with high resistance levels were the most common bacteria causing NIs in the ICU. This study for the first time identified blaOXA-11, blaOXA-23, and blaNDM-1 genes in E. coli and K. pneumoniae in Ilam city of Iran.

Extended-Spectrum β-Lactamase-Producing Escherichia coli and Virulence Genes in Pediatric Patients with Health-Care Urinary Tract Infections.

Healthcare-associated urinary tract infection (UTI) represents a significant health problem, especially in infants and young children. The most common pathogen associated with this infection is Escherichia coli (E. coli). The present study aimed to detect the frequency of virulence genes among clinical isolates of E. coli isolated from healthcare-associated urinary tract infections in children and the correlation between these virulence genes and the presence of the blaCTX gene. The study included one hundred clinical isolates of E. coli isolated from healthcareassociated urinary tract infections in children in intensive care units. The isolates were subjected to antibiotics sensitivity by disc diffusion method and detection of extended-spectrum beta-lactamase by double disc diffusion method. In addition, multiplex polymerase chain reaction (PCR) was used to detect some virulence genes, and PCR was used to detect the blaCTX-M gene. E. coli producing ESBL by double discs method was identified in 74 isolates. blaCTX-M gene detection by PCR was identified among 38 isolates representing 51.4% of ESBL-producing E. coli. There was a significant association between ESBL and blaCTX-M Gene, P = 0.0001. The frequency of the studied virulence genes by multiplex PCR in the isolated E. coli was 66% for the Fim gene, 75% for the Aer gene, 68% for the FliC gene, 53% for each of IucD gene and Usp gene, 40% for pap gene, 35% for each of AFA and ironN genes and 17% for sfa gene. None of the isolated E. coli had the Cdt gene. There was a significant association between the presence of the FimH gene (P = 0.0001), Pap gene (P = 0.05), sfa (P = 0.026), Afa gene (P = 0.018), and aer gene (P = 0.035) and the presence of the blaCTX-M gene in the isolated E. coli. The present study highlights the presence of virulence genes and blaCTX-M gene in uropathogenic E. coli isolated from pediatric patients with healthcare-associated urinary tract infections. There was an association between the blaCTX-M gene and virulence genes FimH, pap, sfa, Afa, and aer. Various distributions of the studied genes with a high frequency of fimbria are flic genes. Moreover, the ESBL had high frequency in E. coli with the presence of blaCTX-M in about one-third of the isolates.

Detection and correlation of antimicrobial resistance pattern in Enterobacteriaceae from UTI patients in GMERS Medical College, Gujarat

Background: Bacterial resistance to antibiotics is a growing public health threat worldwide. The increasing rate of antimicrobial resistance among bacterial pathogens causing both hospital- and community-acquired infections is a serious threat to public health world-wide. This inappropriate and non-judicious usage of antibiotics has resulted in the development of worldwide antibiotic resistance in bacteria, leading to the emergence of multi-resistant strains of bacterial pathogens. This study focuses on the prevalence of antibiotic resistance in the Enterobacteriaceae group of organisms in urine samples and also detects various methods of antibiotic resistance. Antibiotic resistance detection may be useful for epidemiological and research purposes, as well as for preventing the spread of drug-resistant organisms within hospitals through good infection control practices. Aims and Objectives: The aim of the study was to detect occurrence of β-lactamases, extended-spectrum beta-lactamases (ESBL) and Carbapenemase by phenotypic methods in Enterobacteriaceae from urine samples along with pattern of antibiotic resistance for various antibiotics in them. Materials and Methods: A descriptive study was conducted at a tertiary-care hospital. Testing of ESBL and carbapenemase production detection done according to CLSI (M100) guideline by the Kirby-Bauer disk diffusion method, combination disc diffusion test, and modified Carbapenem inactivation method. Results: A total of 220 Enterobacteriaceae organisms were isolated from processed urine samples of tertiary care Hospitals. Rate of cephalosporin resistance in ESBL and carbapenem-resistant Enterobacteriaceae (CRE) is more than 90% while in non-ESBL more than 70% and in non-CRE 75–80%. Carbapenem resistance in ESBL and non-ESBL is the same. Resistance to fluoroquinolone group, Aminoglycoside group, and Cotrimoxazole and Tetracycline group of antibiotics were more noticed in ESBL and carbapenemase producing organisms. In our study, fosfomycin and Nitrofurantoin are effective treatment in case of ESBL and CRE producing organism. Conclusion: The ESBL and Carbapenemase producing isolates were multi-drug resistant making therapeutic choices limited. Fosfomycin and Nitrofurantoin are effective treatment in multidrug resistance urinary tract infection.

EXTENDED SPECTRUM BETA-LACTAMASE (ESBL) PRODUCING PROTEUS SPECIES ISOLATED FROM CLINICAL SPECIMENS FROM SELECTED HOSPITALS IN JIGAWA STATE, NORTH-WEST NIGERIA

Proteus species are found in multiple environmental habitats, including long-term care facilities, hospitals, and can also cause both community and nosocomial infections. For a long time, Proteus was known to be susceptible to beta-lactam antibiotics but nowadays they become resistant. Hence, this study aims to determine the prevalence, antibiotic susceptibility pattern of extended spectrum beta lactamase (ESBL) producing Proteus species across Jigawa State, Northwest Nigeria. 1854 different clinical specimens were analysed of which 191 Proteus species were isolated through standard biochemical tests and used for this study from selected hospital between November, 2021 to August, 2022. Modified Kirby-Bauer disk diffusion method was used to test the susceptibility of the Proteus isolates to nine different antimicrobial agents. Double Disc Synergy Test (DDST) was used for phenotypic detection of ESBL in isolates. The prevalence of ESBL producing Proteus mirabilis was 11.8%. None of the Proteus vulgaris isolates in this study was found to be ESBL producers. Male patients were infected with more ESBL producers than those from female counterparts (Male vs Female; 12.5% vs 4.8%). Proteus mirabilis were observed to be significantly more susceptible to Ampicillin (P= 0.003), Gentamycin (P=0.024) and Cotrimoxazole (P=0.014). All ESBL producing Proteus mirabilis were resistant to three or more classes of antibiotics used in this study. This study reveals the occurrence of ESBL producing Proteus species in this environment. All the ESBL producing Proteus mirabilis encountered in this study exhibited multidrug resistance. Prudent use of antimicrobial agents is advocated in order to tame the trend.

Detection of ESBLs and carbapenemases among Enterobacteriaceae isolated from diabetic foot infections in Ouargla, Algeria.

The emergence and rapid spread of Enterobacteriaceae carrying extended spectrum beta-lactamases (ESBLs) and carbapenemases represent a great threat to clinical treatment due to their multi-drug resistance. This study investigated ESBLs and carbapenemases encoding genes in Enterobacteriaceae collected from diabetic foot infections (DFIs) in Ouargla, southern Algeria. A total of 70 Enterobacteriaceae strains were recovered from 76 patients with DFI between February 2017 and April 2018. Antimicrobial susceptibility testing was performed using the disc diffusion method, and the presence of bla genes was detected using polymerase chain reaction (PCR) and DNA sequencing. The genetic transfer of the plasmids was carried out by conjugation using the broth mating method. The most common isolate was Proteus mirabilis, followed by Escherichia coli, Morganella morganii and Klebsiella pneumoniae. The prevalence of ESBL and carbapenemase-producing Enterobacteriaceae was 11.42% and 2.85 % respectively. Plasmid-mediated AmpC was detected in 5.71% isolates. Conjugation experiments showed the transferability of blaCTX-M-2. Our findings support the view that various pathogens found in DFIs differ from one part of the country to another. This study reports the first description of metallo-β-lactamase NDM-5 producing Klebsiella pneumoniae clinical isolate in Algeria.

Detection Of Extended Spectrum Of Β-Lactamases In Utis Patients At Raipur Chhattisgarh

Enter obacteriaceae families are capable of producing the extended spectrum beta-lactamase (ESBL) enzyme, which results in highlevels of resistance to all beta-lactam antibiotics. For effective antibiotic therapy, to stop the spread of resistance mechanisms, and forepidemiological reasons, their detection is crucial. 53 Gram negative isolates were detected using a standard methodology in the currentinvestigation, which compared various phenotypic methods for ESBL detection. Antibiotic susceptibility testing (AST) was alsoperformed on these isolates. Phenotypic confirmatory Double Disk Synergy Test (30 mm), Double Disk Synergy Test (30 mm), ModifyDouble Disk, and the ESBL E-Test were used to further validate the presence of ESBLs. 20 ESBLs were produced in the case of DDST(30 mm), and specificity for cefepime was 84%, rising to 86% in the case of cefepime combination. The sensitivity was high forCefepime alone (92%), while in combination it was 87% and 92% with Ceftazidime and Cefotaxime. DDST (20mm) 15 ESBLproductions was seen once more. MDDST (20mm) has a sensitivity of 94% and identified 20 isolates as ESBL. The generation of 20ESBLs was seen in the ESBL E test, and Cefepime alone showed the highest level of sensitivity. As a result, it was discovered thatcefepime was the most effective cephalosporin for ESBL detection, followed by cefotaxime, ceftazidime, and cefpodoxime.

Detection of ESBL producing E.coli and Klebsiella spp in urinary tract infection due to long exposure of catheterize condition in Covid positive patient admitted in HDU

Long exposure of broad spectrum antibiotic can severe causes of extended-spectrum beta-lactamase (ESBL) production within bacteria in UTI patient. Community-acquired infections due to ESBL producing and both are rising worldwide. To assess the prevalence and antibiotics resistance patterns of ESBL-producing and Klebsiella spp. from covid positive patient admitted in HDU with catheterized condition. A hospital-based cross-sectional study was conducted on Salboni Super speciality Hospital (Covid Level-IV) at West Bengal, India from August 2020 to August2021and a total of 1116 urine samples were collected from catheterized patient admitted in male and female HDU due to exposure of covid. Samples were cultured on CLED agar and then Identification of the isolates was done by Gram's staining followed by biochemical tests. Antibiotic susceptibility testing was done by modified Kirby-Bauer disc diffusion method and interpretation was done following Clinical and Laboratory Standard Institute (CLSI) guidelines foe detection of ESBL producing and . present study showed 51.6% (n-326) and 37.2% (n-235) Klebsiella spp. from 631 significant growths from both male and female patients. Out of this, total ESBL producing and spp. seen 55% (n-309) from both male and femle patient in different age group. Finally Isolated ESBL producing i 45.9 % (n-142) and ESBL producing Klebsiella spp. 54 % (n- 167) from both male and female HDU.Detection of ESBL producing and due to prolonged uses of broad spectrum antibiotic vigorously. The occurrence of multidrug resistance to the third-generation cephalosporins, aminoglycosides, fluoroquinolones, trimethoprim-sulfamethoxazole, and tetracyclines is more common among ESBL producers. It may help to detect other organism in same condition, so that it will be importance in clinical aspect of future treatment for clinician.

Biofilm formation by ESBL-producing uropathogenic E. coli

Introduction: E. coli is considered to be the leading cause of urinary tract infections. (UTIs) and is responsible for both community acquired and hospital acquired UTIs. The ability of this bacterium to form biofilm on various devices like catheters plays an important role in progression of the disease. Biofilm production in extended spectrum beta lactamases (ESBLs) producing organisms provides a favourable environment for the exchange of resistance genes and may also facilitate widespread dissemination.
 Methods: The urine samples were collected from the suspected cases of urinary tract infections and were cultured on cystine lactose electrolyte deficient (CLED) agar plates to isolate the desired UPEC strains. All the isolated E. coli strains were subjected to ESBL detection by double disk approximation method and to biofilm formation by tissue culture plate method of biofilm detection. All the results were statistically analysed using k2 test with p value at <0.05 level of significance.
 Results: Out of all the gram negative bacilli isolated from urine samples, E. coli alone accounted for 90% of the isolates. Of all the UPEC isolates subjected to ESBL detection 50% of the strains turned out to be the ESBL producers and 35% of the strains showed biofilm formation.
 Conclusion:
 
 The majority of the work in literature sites the data samples collected from indoor patients, where these isolates coli strains which produced ESBLs were mainly a strong biofilm former as compared to non-ESBL producer strains.
 In this study in spite of all the coli strains being outpatient isolates still the correlation between ESBL production and biofilm formation holds true.
 We found a significant correlation between biofilm formation and ESBL production. The detection of ESBL production and biofilm formation pattern will help in development of newer and more effective treatment.

Molecular Detection of Extended Spectrum Beta-Lactamases in Clinical Isolates of Pseudomonas aeruginosa

Pseudomonas aeruginosa producing extended spectrum beta lactamases (ESBL) is a major concern in the hospital settings. It is usually reported in Enterobacteriaceae and is less frequently observed in P. aeruginosa. There is no recommended test for ESBL detection in P.aeruginosa. Therefore, we determined the occurrence of ESBL in clinical isolates of P.aeruginosa by both phenotypic and genotypic methods. Antimicrobial susceptibility tests were done on two hundred and thirteen isolates of P. aeruginosa. Phenotypic detection of ESBL was performed using combined disk method and ESBL encoding genes such as blaVEB, blaPER, blaPSE, blaGES, blaTEM, blaSHV, blaCTX-M, blaBEL, blaOXA1, blaOXA10, blaOXA2 were studied by simplex PCR. Of the 213 isolates, 85 were identified as resistant to ceftazidime and 27/85 isolates were confirmed to be ESBL producers by phenotypic method. The presence of genes encoding ESBLs comprising of blaTEM (n=44), blaOXA-10 (n=19) isolates, blaOXA-1 (n=5), blaOXA-2 (n=3) were found. All OXA gene positive isolates exhibited the ESBL phenotype. The blaGES gene were identified in 4/85 (5%) isolates. This study shows the prevalence of ESBL among clinical isolates of P.aeruginosa and in particular, the presence of GES β lactamases.

Assessment of microbial quality and detection of extended spectrum β-lactamase genes in Gram-negative bacterial isolates of herbal mixtures commonly hawked in Sagamu metropolis, Ogun State, Nigeria

Background: The use of herbal mixtures in the treatment of diseases is on the rise. Many of these herbal drugs are not produced under hygienic conditions and safety issues associated with herbal medicines may have an exacerbated impact in immunocompromised and elderly individuals. This study aimed to determine the microbial loads of locally prepared herbal mixtures and detect extended spectrum beta-lactamase (ESBL) genes in any isolated Gram-negative bacteria pathogen.
 Methodology: Fifty local herbal mixtures were purchased randomly from three locations in Sagamu town (Sagamu market, Ita-Oba Road and Isale Oko) in Ogun State, Nigeria. The mean total viable bacterial (MTVB), mean total coliform (MTC), and mean total fungal (MTF) counts were determined by the plate count method. The bacterial isolates were streaked on differential bacteriological media while the fungi isolates were grown on potato dextrose agar. The isolates were identified upon growth on culture media using conventional biochemical tests. Antibiotic susceptibility pattern of the isolates was determined using Kirby-Bauer disk diffusion technique. Phenotypic detection of ESBL was done by the modified double disc synergy test followed by amplification detection of blaTEM, blaCTX-M and blaSHV genes with polymerase chain reaction (PCR) assay.
 Results: Bacteria and fungi were isolated from 38 (76.0%) and 25 (50.0%) of the herbal samples respectively. Ten (20.0%) and 14 (28.0%) of the samples had mean bacterial and fungal load that exceeded 105CFU/mL or g, respectively. Nineteen (38.0%) of the herbal samples analyzed had total coliforms. Fifty-one isolates belonging to eight bacterial genera and 28 fungi isolates belonging to four fungal genera were obtained. Thirty- two (62.7%) of the bacterial isolates were Gram-negative while 19 (37.3%) isolates were Gram-positive. Staphylococcus aureus was the most common bacterial isolate (33.3%) while Aspergillus species was the most prevalent fungus (60.7%). Sixteen (84.2%) S. aureus and 26 (81.3%) Gram- negative isolates were multidrug resistant, and 6 (18.8%) of 32 Gram-negative isolates were ESBL producers. ESBL-encoding genes were detected in 7 (27%) of the 26 multidrug resistant Gram-negative bacteria with TEM and SHV being the most prevalent 4 (14.8%) while CTX-M was identified in only one isolate.
 Conclusion: This study reported the presence of microbial contaminants which exceeded the safety limits of 105 CFU/g according to World Health Organization. The use of locally prepared herbal medicines poses a major health risk due to the lack of microbial quality standards.

Detection of extended spectrum beta-lactamase (ESBL)–production in Salmonella Typhimurium isolated from poultry birds in Nasarawa State, Nigeria

Salmonella Typhimurium is an important zoonotic food borne pathogen and the second most widely studied pathogen in relation to antimicrobial resistance studies in Nigeria. Its resistance to β-lactam antibiotics is on the rise globally due to persistent exposure and production of Extended Spectrum Beta-lactamase enzymes (ESBLs) which is barely reported to occur among food producing animals in Nigeria. This poses a threat to the successful therapy of salmonellosis infections and raises concern on food safety. Further complicating these challenges is the lack of strict laws mitigating antibiotic usage in Nigeria. The study was conducted in the Microbiology laboratory of Nasarawa State University, Keffi, Nigeria. A total of 300 samples each of poultry droppings, flesh, feeds, human hand swabs and stool were randomly collected following ethical approval and screened for ESBL- producing S. Typhimurium using standard Microbiological methods of isolation and identification. Antibiotic susceptibility testing, phenotypic ESBL detection and interpretation were carried out as described by the Clinical Laboratory and Standard Institute (CLSI) guideline. The bla genes were identified using polymerase chain reaction (PCR) method. A total of 106 S.Typhimurium were isolated with the following resistance profile; Ampicillin 99(93.4%), Augmentin 104(98.1%), Ceftriaxone 74(69.8%), Ceftazidime 50(47.2%), Imipenem 1(1.0%). The antibiotic resistance of S. Typhimurium in the study is significant (P = 0.01). The Multiple Antibiotic Resistance (MAR) Index was > 0.2. Out of 106 isolates, 32 distinct antimicrobial resistant phenotypes were observed and 62 (81.6%) were confirmed ESBL producers having 19 distinct phenotypes. The bla genes were identified among 38.7% (24/62) isolates; 100% blaSHV (24/24), 87.5 %(21/24)blaTEM with 54.2% (13/24) co-carriage of blaTEM-SHV. The blaCTX-M was not detected. Poultry is therefore an important reservoir and source of transmission of ESBL- producing S.Typhimurium, which is a major public health concern. The high MAR index suggests frequent antibiotic usage within our study area, thus calls for antibiotic stewardship in animal production in Nigeria.