Extended-spectrum Beta-lactamase Screening Research Articles

EXTENDED SPECTRUM BETA LACTAMASE PRODUCING UROPATHOGENS IN A TERTIARY CARE HOSPITAL IN SOUTH INDIA: A CLINICO MICROBIOLOGICAL STUDY

Introduction: Urinary Tract Infection (UTI) is the most common bacterial infection encountered in both hospital and community setup and the presence of drug resistance carries remarkable clinical implication. Drug resistant phenotypes like ESBL (Extended Spectrum Beta lactamase producers) are difcult to treat causing increased morbidity. Aims: The present study aims to determine the prevalence of ESBL producing Enterobacteriacea among Uropathogens and to determine the Antibiotic susceptibility pattern. Materials and Methods :A total of 726 urine samples were obtained and processed as per standard microbiological techniques .Antibiotic susceptibility testing was done by Kirby Bauer disc diffusion method. All the isolates were screened for ESBL production and conrmed by Double Disc Synergy test (DDST).Differences between the antibiotic susceptibility pattern ,Various risk factors were assessed among the ESBL producing and non ESBLproducing bacteria and the signicance was analyzed . Statistical analysis used:Chi square test.(p < 0.05 signicant) Results: A total of 726 urine samples were processed of which 421 bacteria belonging to Enterobacteriaceae were included in the study. Most common organism isolated was Escherichia coli 279 (66.27%).Highest susceptibility was observed for Imipenem 361 (85.7%) and Amikacin 356 (84.5%).Out of 461 isolates, 235 (55.8%) isolates were positive for ESBL screening all the isolates were conrmed with DDST. There was a signicant difference for Diabetes mellitus, CAUTI and Inpatients among the antibiotic susceptibility pattern between ESBL and non ESBL producers.(p<0.0001) Conclusions: Early detection of Multidrug resistance isolates like ESBLproducers in routine testing could help avoid treatment failure. Screening for ESBL production needs to be carried out regularly in every clinical diagnostic laboratory to guide clinicians in proper selection of antibiotics. Continued monitoring of the susceptibility pattern of ESBLproducing bacteria will provide valuable information regarding the local antimicrobial susceptibility pattern which in turn will guide in choosing the initial empirical therapy

Risk factors associated with faecal carriage of extended-spectrum cephalosporin-resistant Escherichia coli among dogs in Southeast Brazil

Faecal carriage of extended-spectrum cephalosporin-resistant Escherichia coli (ESC-R E. coli) in dogs has been reported worldwide and can reduce the effectiveness of treatments against bacterial infections. However, the drivers that influence faecal carriage of ESC-R E. coli in dogs are poorly understood. The aims of this study were to estimate the prevalence of ESC-R E. coli among dogs prior to their admission to a veterinary teaching hospital and to identify risk factors associated with the faecal carriage of ESC-R E. coli. Rectal swabs (n = 130) were collected from dogs and screened for ESC-R E. coli using MacConkey agar supplemented with cefotaxime (2 μg/mL). E. coli species was confirmed by MALDI-TOF and screening of extended-spectrum beta-lactamase (ESBL) genes was conducted by multiplex PCR. Questionnaires were completed by each dog’s owner to test several human and dog characteristics associated with ESC-R E. coli. The prevalence of faecal carriage of ESC-R E. coli was 9.2 % and 67 % of ESC-R E. coli isolates harboured ESBL genes including CTX-M alone or in combination with TEM. All ESC-R E. coli isolates were resistant to ceftriaxone, cefpodoxime, and cefotaxime and were susceptible to cefoxitin and carbapenems. The likelihood of carrying ESC-R E. coli was 15 times higher (OR = 14.41 [95 % CI: 1.80−38.02], p < 0.01) if the dog was treated with antibiotics 3–12 months prior to sampling and 8 times higher (OR = 7.96 [95 % CI: 2.96−92.07], p < 0.01) if the dog had direct contact with livestock, but 15 times lower (OR = 0.07 [95 % CI: 0.01−0.32], p < 0.01) if the dog was dewormed during the previous year. Our findings confirm the faecal carriage of ESC-R E. coli in subclinical dogs and call for further investigation regarding the impact of deworming on antibiotic-resistant bacteria in companion animals.

The Escherichia coli Sequence Type 131 Harboring Extended-Spectrum Beta-Lactamases and Carbapenemases Genes from Poultry Birds.

Background and AimThe extended-spectrum beta-lactamases (ESBLs), as well as carbapenemases, are considered as the foremost resistance determinants throughout the world. However, the relevant data especially related to the sequence types of ESBL and carbapenemases producing Escherichia coli from the poultry is limited from Pakistan. Here, we present the data on the genetic diversity of E. coli strains isolated from the poultry birds from the poultry farms located in Islamabad, Pakistan, and the underlying resistance mechanisms to beta-lactam agents.MethodsOf 250 broilers from 25 different farms (10 birds from each farm), the cecal samples were obtained and analyzed for the presence of ESBLs producing E. coli (ESBL-Ec) as well as carbapenemases producing E. coli (CPEc) strains using selective agar for ESBL and carbapenemases screening. The susceptibility profiling of the ESBL-Ec and CPEc isolates was evaluated followed by multi-locus sequence typing.ResultsA total of 119 strains were positive for ESBL production whereas 37 strains were found positive to produce carbapenemases in addition to ESBLs. The MLST analysis has shown a diversity of isolates as the E. coli isolates from poultry birds correspond to a total of 16 sequence types (STs). The ST131 (22/48, 46%) followed by ST8051 (10/48, 21%) were the main STs in this study. The blaCTX-M gene was detected in all the poultry E. coli strains whereas the blaTEM was found in 45.5% of strains. The blaVIM was found in all 37 CPEc isolates whereas the blaNDM and blaIMP were found in 31/37 (83.8%) and 16/37 (43.2%) CPEc isolates respectively.ConclusionThe overall results have shown the prevalence of diverse genotypes among the ESBL-Ec and carbapenemase-producing E. coli (CPEC) from poultry. Furthermore, the study documents poultry birds as a persisting reservoir of extensively antimicrobial-resistant E. coli ST131 in Pakistan, suggesting a potential threat to public health.

1267. Comparative activity of omadacycline against extended-spectrum beta-lactamase positive and negative Escherichia coli and Klebsiella pneumoniae strains recovered from urine specimens

BackgroundOmadacycline (OMC) is a novel tetracycline (TET) derivative antibiotic with activity against TET-resistant Enterobacterales. OMC is available in both oral and intravenous formulations and is has been studied as a treatment of uncomplicated urinary tract infection (UTI) and acute pyelonephritis. The purpose of this study was to evaluate OMC activity against extended-spectrum beta-lactamase (ESBL) positive and negative Enterobacterales strains recovered from urine specimens.MethodsUrine samples from patients with suspected UTI were quantitatively plated onto blood agar and MacConkey agar plates in the microbiology lab of Wake Forest Baptist Medical Center. After overnight incubation, colonies were identified to the species level by MALDI-TOF system. Susceptibility testing was performed for isolates of E. coli and K. pneumoniae. OMC and TET susceptibility testing was performed by disk diffusion and gradient strip methodologies. Results were interpreted in accordance with the Clinical and Laboratory Standards Institute (CLSI) or Food and Drug Administration breakpoints. Isolates were tested in triplicate. ESBL screening and susceptibility testing to oral antibiotics commonly prescribed for UTI were performed by the MicroScan WalkAway System. Susceptibility rates and MIC50/90 were calculated and subsets of isolates were analyzed using descriptive statistics.ResultsA total of 204 isolates, including 102 E. coli and 102 K. pneumoniae, were tested. All but 1 isolate (99.5%) exhibited categorical agreement in results generated by the strip (Table 1) and disk (data not shown) methods and this was considered a minor error involving an intermediate result. OMC MIC90 for E. coli and K. pneumoniae were 6 µg/mL and >32 µg/mL, respectively. OMC displayed increased susceptibility rates compared to TET regardless of isolate species or ESBL positivity (Table 2).Table 1. Omadacycline Minimum Inhibitory Concentrations (MICs, µg/mL) Table 2. Susceptibilities of Oral Antibiotics Used to Treat UTI (% S) ConclusionOMC exhibits promising antimicrobial activity against TET-resistant and ESBL-positive E. coli and K. pneumoniae. OMC displays superior activity to ESBL positive E. coli when compared to ESBL positive K. pneumoniae. These data support the development of OMC as a much needed option in the treatment of UTI caused by resistant Enterobacterales.Disclosures Tyler J. Stone, PharmD, Paratek (Research Grant or Support) Abdullah Kilic, MD, Paratek (Grant/Research Support) John Williamson, PharmD, Paratek (Research Grant or Support) Elizabeth Palavecino, MD, Paratek (Grant/Research Support)Paratek (Grant/Research Support)

A Comparative In Vitro Sensitivity Study of "Ceftriaxone-Sulbactam-EDTA" and Various Antibiotics against Gram-negative Bacterial Isolates from Intensive Care Unit.

IntroductionA rapid increase in multidrug-resistant (MDR) strains is being seen across the globe especially in the Southeast Asian region, including India. Carbapenems and colistin form the mainstay of treatment against gram-negative pathogens, especially extended-spectrum beta-lactamase (ESBL)- and metallo-beta-lactamse (MBL)-producing isolates. However, due to increased resistance to carbapenems and toxicity of colistin, especially in intensive care units (ICUs), carbapenem-sparing antibiotics like ceftriaxone–sulbactam–EDTA (CSE) combination needs to be evaluated.Materials and methodsBacterial isolates cultured from various clinical samples from all ICUs for a period of 9 months were evaluated. Bacterial identification was performed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF MS) and antibiotic susceptibility testing were performed by disk diffusion and E test method. Antibiogram of various antibiotics was noted. Extended-spectrum beta-lactamase- and MBL-producing bacteria were identified by phenotypic methods. Antibiotic sensitivity results of CSE were compared with the comparator drugs like colistin, carbapenems, and tigecycline in Enterobacteriaceae, Acinetobacter spp., and Pseudomonas spp. along with ESBL and MBL producers.ResultsA total of 2,760 samples of blood, cerebrospinal fluid (CSF), respiratory samples, tissue, and pus were collected from ICUs with maximum isolates from pus (37%) followed by respiratory samples (31%) and blood (27%). Escherichia coli and Klebsiella pneumoniae were the predominant gram-negative pathogens accounting for 56% of the isolates followed by Acinetobacter spp. (23%) and Pseudomonas spp. (15%). Extended-spectrum beta-lactamase screening was positive for 57% (1,069/1,877) isolates; whereas 43% (732/1,877) were MBL producers. According to the antibiotic susceptibility results, CSE was the most effective antibiotic showing 94% sensitivity for carbapenem-sensitive Enterobacteriaceae and 97% for carbapenem-resistant Acinetobacter and Pseudomonas spp. Among the other drugs, colistin was found to be the most effective showing almost 95% sensitivity in both the Enterobacteriaceae and non-Enterobacteriaceae group (both ESBL + OXA/NDM). Ceftriaxone–sulbactam–EDTA was also found much more effective (95%) as compared to Colistin (89%) toward ESBL- and MBL-producing strains of Enterobacteriaceae and non-Enterobacteriaceae group. Among the carbapenems, imipenem was the most effective drug against Enterobacteriaceae showing 34% sensitivity and ertapenem proved to be least effective.ConclusionIn our present study, CSE emerged as a potent antibacterial agent against MDR gram-negative infections; both for ESBL as well as MBL producers. Hence, in light of present study, we strongly recommend inclusion of CSE in routine sensitivity panel and may be used as a carbapenem- and colistin-sparing drug and a promising option against ESBL and MBL producers especially in ICU.How to cite this articleSingh S, Sahu C, Patel SS, Singh A, Yaduvanshi N. A Comparative In Vitro Sensitivity Study of “Ceftriaxone–Sulbactam–EDTA” and Various Antibiotics against Gram-negative Bacterial Isolates from Intensive Care Unit. Indian J Crit Care Med 2020;24(12):1213–1217.

Prevalence of multidrug-resistant and extended-spectrum beta-lactamase (ESBL)-producing gram-negative bacilli: A meta-analysis report in Ethiopia.

Multidrug-resistant (MDR) extended-spectrum beta-lactamase (ESBL)-producing bacterial isolates have emerged as a global threat to human health. Little is known about the overall prevalence of multidrug resistance profile and ESBL-producing gram-negative bacilli (GNB) in Ethiopia. Therefore, this meta-analysis was performed to produce proportional estimates of multidrug resistance and ESBL-producing GNB in Ethiopia. A web-based search was conducted in PubMed, Google Scholar, Research Gate, Scopus and other databases. Articles published till 2019 on the prevalence and antimicrobial resistance profiles of ESBL-producing GNB in Ethiopia were included in the study. Relevant data were extracted and statistical analysis was performed using comprehensive meta-analysis version 3.3.0 software. Publication bias was analyzed and presented with funnel plots. In this meta-analysis, the overall proportional estimate of ESBL-producing GNB was 48.9% (95% confidence interval [CI]: 0.402, 0.577). The pooled proportional estimates of ESBL-producing Klebsiella pneumoniae, Escherichia coli and other GNB were 61.8%, 41.2% and 42.9%, respectively. Regarding antimicrobial resistance profiles against selected drugs, the pooled proportional estimates of resistance against amoxicillin-clavulanic acid, trimethoprim-sulfamethoxazole, cefotaxime, ceftazidime, tetracycline, gentamicin and ciprofloxacin was 79.0%, 78.4%, 78.0%, 72.4%, 72.7%, 58.9% and 43.8%, respectively. The pooled proportional estimates of MDR isolates were found to be 82.7% (95% CI: 0.726, 0.896), which are relatively high as compared to other countries. This highlights a need for active surveillance systems which can help understand the actual epidemiology of ESBL, aid in formulating national guidelines for proper screening of ESBL and support developing standardized approaches for managing patients colonized with ESBL.

Clinical and drug resistance characteristics of Providencia stuartii infections in 76 patients.

ObjectivesTo investigate the clinical and drug resistance characteristics of Providencia stuartii infections in the Huainan region of Anhui and provide a reference for the clinical selection of antimicrobial agents.MethodsThis single-center retrospective analysis included 76 patients with P. stuartii infection in Huainan during the period from October 2018 to March 2020. The hospital department in which the patients were treated and the drug susceptibility characteristics of the P. stuartii isolates were recorded.ResultsAmong the 76 patients, the lung was the most common site of infection, and intensive care unit was the main hospital department. Extended spectrum beta-lactamase screening revealed expression by all 76 isolates of P. stuartii. Of the 76 isolates, 92.1% exhibited multiple drug resistance or extensive drug resistance. P. stuartii isolates were sensitive to cefepime and imipenem, but not to other beta-lactam antibiotics. Twenty isolates were resistant to all 21 types of antibiotics. Of the 20 patients infected with extensively drug-resistant isolates, nine (45%) died.ConclusionsDrug resistance is increasing in P. stuartii. The antimicrobial agent imipenem may be effective for treatment of P. stuartii infections. Fluoroquinolones, aminoglycosides, and fourth-generation cephalosporins are suitable options for antibiotic therapy.

Microbial and Parasitic Contamination of Fresh Raw Vegetable Samples and Detection of the BlaTEM and BlaCTX-M Genes from E. coli Isolates

A total of 100 fresh-raw vegetable samples were collected from the Kathmandu, Lalitpur and Bhaktapur districts of Nepal to evaluate microbial and parasitic contamination, presence of Extended Spectrum Beta Lactamase (ESBL)-producing Escherichia coli and detect the blaTEM and blaCTX-M genes among the Escherichia coli isolates. This study revealed that the prevalence of Giardia cysts was highest (100%) and Hookworm and Entamoeba coli were lowest (1% each). Coliforms were isolated from every raw vegetable sample. A total of 178 bacterial isolates were isolated among which 57 isolates were identified as E. coli, out of which 33 were Multi-drug resistant (MDR) isolates. The high rate of resistance was found towards amoxicillin/clavulanate, tetracycline and cotrimoxazole. The 10 E. coli isolates tested positive in an ESBL screening, out of which 4 were confirmed as ESBL producers by a combined disc test. Out of these 4 confirmed ESBL E. coli, one was found to carry both the blaTEM gene and blaCTX-M genes by the Polymerase Chain Reaction (PCR) technique. One isolate has only the blaTEM gene, while other isolate harboured only blaCTX-M genes.

Screening for urinary tract infection-causing Klebsiella pneumoniae using CHROMagar

Introduction. Urinary tract infections (UTIs) are considered prevalent among humans. While Klebsiella pneumoniae and Escherichia coli are commonly isolated from patients with UTIs, K. pneumoniae is more frequently isolated from specimens isolated from hospital-acquired infections. Aim. The current study aimed to characterise and evaluate pathogenic K. pneumoniae isolated from patients suspected with a UTI at King Abdulaziz University Hospital. Methodology. Twenty-four urine samples obtained from patients, between 12 November 2018 and 11 January 2019, were included in the study, and the microbial content was analysed via urine culture and VITEK 2 analyses. Results. Of the 24 urine specimens, 23 samples (95.8%) yielded significant microbial growth (&gt; 105 CFU/mL K. pneumoniae), and one sample (4.2%) yielded non-pathogenic microbial growth (&lt; 105 CFU/mL K. pneumoniae). Of the specimens with cell counts of &gt;105 CFU/mL, 21 samples (87.5%) were pure cultures showing the growth of a single pathogen, and three samples (12.5%) were mixed cultures, showing the growth of two or more pathogens; 10 (41.7%) samples were extended-spectrum beta-lactamase (ESBL) producers. VITEK 2 analysis showed that the most effective antibiotic was piperacillin, with 87.5% of the strains isolated in this study showing sensitivity to it, followed by gentamicin (83.3%) and ciprofloxacin (79.2%). Antibiotic susceptibility studies identified resistance against ampicillin (45.83%), trimethoprim (41.67%), and amoxicillin (25%) in the study isolates. Differential chromogenic culture media CHROMagar ESBL and CHROMagar KPC (K. pneumoniae carbapenemase) were used for rapid screening of ESBL and KPC producers. Among the 24 K. pneumoniae isolates, six isolates (25%) formed metallic blue colonies on CHROMagar ESBL, five (20.3%) were inhibited, and nine (37.5%) showed weak pigmentation. Three isolates (12.5%) formed pink colonies on CHROMagar ESBL. Seventeen isolates (70.8%) were KPC-positive, four (16.7%) showed weak pigmentation, and three (12.5%) formed pink colonies on CHROMagar KPC. The results from CHROMagar cultures agreed with those of VITEK 2 analyses. Conclusion. Various microbial detection methods have been used in medical microbiology laboratories to identify and screen for microbial resistance in clinical specimens. VITEK 2 and CHROMagar are amongst the commonly used methods. Here, we report a possible discrepancy between the detection techniques and that the data obtained by different methods need not be synergistic. However, the use of a chromogenic medium offers a quick and accurate method for detection, enumeration, and presumptive identification of urinary tract pathogens.

Phylogenetic Characterization and Multilocus Sequence Typing of Extended-Spectrum Beta Lactamase-Producing Escherichia coli from Food-Producing Animals, Beef, and Humans in Southwest Nigeria

Multidrug-resistant extended-spectrum beta lactamase (ESBL)-producing Escherichia coli strains are emerging globally in both humans and animals. Antimicrobial susceptibility testing and ESBL screening were performed on pure cultures of 216 E. coli isolates from human and animal fecal samples as well as beef. Polymerase chain reaction was performed for the detection of resistance genes. Representative isolates of ESBL-producing E. coli were randomly selected for multilocus sequence typing and pulsed field gel electrophoresis (PFGE). Sixty of the isolates were identified as ESBL producers, and seven resistance genes were amplified in them: TEM (61.7%), blaCTX-M-15 (51.7%), AAC-6-LB (43.3%), blaCTX-M-1 (38.3%), blaCTX-M-9 (33.3%), blaCTX-M-2 (21.7%), and SHV (11.7%); they were classified into four phylogroups: A (25%), B1 (45%), B2 (20%), and D (10%). Thirty of these isolates were clustered into 10 sequence types with ST131 being mostly prevalent. Six PFGE types were discovered, each of which was shared by isolates from different subjects and had the same phylogroups and resistance gene profiles. There was a dissemination of PFGE types across various groups among humans, animals, and beef. This underlines the fact that the spread of ESBL E. coli could be from humans to animals, from animals to humans, as well as across animal species.

Extended spectrum beta lactamase (ESBL)-producing Klebsiella pneumoniae clinical isolates and its susceptibility pattern to antibiotics at Dr. Soeradji Tirtonegoro General Hospital Klaten, Central Java

Globally, the prevalence of Klebsiella pneumoniae (K. pneumonia) producing extended spectrum beta lactamase (ESBL) has been increasing steadily. The susceptibility patterns of ESBL-producing K. pneumonia varies considerably among countries. Therefore, the investigation of ESBL-producing K. pneumoniae in clinical isolates and their susceptibility are warranted. This research aimed to determine the proportion of ESBL-producing K. pneumoniae and the antibiotic susceptibility patterns of clinical isolates from Dr. Soeradji Tirtonegoro General Hospital, Klaten, Central Java. Identification of K. pneumoniae was performed by analyzing colony morphology, microscopic examination, and biochemical testing using Microbact. Both antibiotic susceptibility testing and ESBL screening (using ceftazidime, cefotaxime, and ceftriaxone discs) were conducted using disc diffusion method according to CLSI. The positive results were confirmed with modified double disk synergy (MDDST) using amoxicillin-clavulanate, ceftazidime, cefotaxime, and cefepime discs. From 962 clinical bacterial isolates, 168 (17.46%) isolates were identified as K. pneumoniae, during June 2017-May 2018. K. pneumoniae was mainly isolated from the Intensive Care Units (ICU) (29.17%) and with sputum being the most common specimen (45.24%). Overall ESBL producers were 52.98%, with the majority from ICU (41.57%) and isolated from sputum specimens (40.45%). ESBL-producing K. pneumoniae showed high resistance to many antibiotics. The sensitivity of ESBL-producing K. pneumoniae isolated from respiratory tract samples against piperacillin-tazobactam, amikacin, and meropenem was more than 80%. In conclusion, among all K. pneumoniae isolates, ESBL K. pneumoniae was 52.98%. ESBL K. pneumoniae from respiratory tract specimens had a sensitivity of more than 80% against piperacillin-tazobactam, amikacin, and meropenem.

Implications of faecal ESBL carriers undergoing TRUS-guided prostate biopsy (TRUSPB): role of screening prior to TRUSPB.

Modification of antibiotic prophylaxis prior to transrectal ultrasound-guided prostate biopsy (TRUSPB) based on pre-procedure rectal culture results is effective for prevention of infection from fluoroquinolone (FQ)-resistant and extended-spectrum beta-lactamase (ESBL) Escherichia coli strains. This has several implications for service delivery and cost. Our aim was to audit sepsis rates after introduction of ESBL screening and to identify risk factors for FQ resistance in ESBL strains and factors for sepsis risk. This was a prospective cohort study from 2013 to 2016. TRUSPB patients underwent pre-procedure rectal swabs. ESBL-positive patients received amikacin in addition to ciprofloxacin prophylaxis. Patients filled a formal risk assessment questionnaire prior to biopsy. Sepsis rate after introduction of targeted prophylaxis reduced from 3.1% (2009-2012) to 1.4% (2013-2016). Of 38 ESBL patients, n = 5 (13%) developed severe post-TRUSPB sepsis. Among the FQ-resistant ESBL producers, the sepsis rate was 24%. Predictive factor for FQ resistance in ESBL producers included-antibiotic use in the last 3months (OR 15). The logistic regression analysis did not identify any significant factor for post-TRUSPB sepsis in ESBL-positive patients once they had received additional prophylaxis. In the face of rising TRUSPB sepsis and higher sepsis rates with ESBL carriers despite additional prophylaxis, introduction of a targeted antibiotic prophylaxis prior to TRUSPB sepsis prostate biopsies based on rectal swabs or urine cultures may reduce sepsis rates or clinicians may find themselves leaning towards increasingly performing transperineal biopsies with lower sepsis rates supporting the 'trexit' initiative.

Extended Spectrum Beta-lactamase Producing Gram Negative Bacterial Isolates from Urine of Patients Visiting Everest Hospital, Kathmandu, Nepal

Objectives: The study was aimed to determine the prevalence of Extended Spectrum Beta Lactamase (ESBL) producing Gram negative pathogens from urine samples along with their antimicrobial resistance.&#x0D; Methods: This cross-sectional study was conducted from December 2015 to May 2016 at Everest Hospital, Kathmandu. Mid-stream urine samples were collected and processed for culture by standard loop streak method. Identified bacterial isolates were tested for Antibiotic Susceptibility by modified Kirby Bauer disc diffusion method and, were subjected to ESBL screening by using 30µg cefotaxime and ceftazidime. ESBL production was confirmed by combination disc method.&#x0D; Results: Of the three hundred urine samples, 22.7% (67/300) showed significant growth. Four different bacterial species were identified. Among the isolates, E. coli was the most common pathogen (71.64%) followed by Klebsiella pneumoniae (14.92%), Pseudomonas spp (8.95%) and Acinetobacter spp (4.48%). Altogether 92.54% (n=62) isolates were sensitive to gentamicin, 89.55% (n=60) to amikacin, and 79.10% (n=53) to nitrofurantoin. 70.10% (n=47) isolates were resistant to antibiotic ampicillin while 62.68% (n=42) were found as Multi-Drug Resistant (MDR) and 29.8% (n=20) were ESBL producers.&#x0D; Conclusions: The overall prevalence of MDR and ESBL among uropathogens is low in comparison to other studies though it is essential to have a regular monitoring of ESBL producing clinical isolates in laboratory practice.

English

This study investigated the co-carriage of plasmid mediated quinolone resistance (PMQR) and extended spectrum beta-lactamase (ESBL) producing lactose non-fermenting (LNF) Enterobacteriaceae isolated from poultry birds. This was a descriptive cross-sectional study carried out between September, 2016 and March, 2017. The Kirby-Bauer disk diffusion method was used to determine the antimicrobial susceptibility patterns. ESBL screening disc kit was used to detect ESBL activities. Detection of ESBL and PMQR genes was carried out by means of polymerase chain reaction. In total, 207 LNF Enterobacteriaeae isolates were recovered from the cloacal swabs of poultry birds within the Calabar Metropolis. ESBL-producing isolates were 162 (78.3%) while fluroquinolone resistant isolates were 194 (93.7%). Among the ESBL-producing isolates, resistance to Ciprofloxacin, Norfloxacin, Levofloxacin, Ofloxacin and Nalidixic acid was 55 (34.2%), 26 (16.1%), 35 (21.7%), 50 (31.1%), and 162 (100%), respectively. About 65% of the quinolone resistant isolates were positive for at least one of the PMQR and ESBL genes in this study. Strict antimicrobial screening, surveillance of resistant isolates as well as the judicious practice of antimicrobial administration in the poultry setting with special emphasis on fluoroquinolones is advised given the high prevalence of co-existent ESBL and PMQR genes. Key words: LNF enterobacteriaeae, Extended spectrum beta-lactamases, quinolone resistance

The Incidence of Extended-Spectrum Beta-Lactamase (ESBL)-Producing Bacteria in Salad Vegetables in Ondo City, Nigeria

Aim: This study was carried out to determine the occurrence of extended-spectrum beta-lactamase (ESBL) producing bacteria in salad vegetables in Ondo City, Nigeria.&#x0D; Study Design: An experimental study design with randomized sampling.&#x0D; Place and Duration of the Study: The research was carried out in the Department of Biological Sciences of Wesley University, Ondo State, Nigeria.&#x0D; Methodology: Samples of cucumber, carrot, green pea, green beans, sweet corn and cabbage were analysed on appropriate agar medium. Pure isolates were identified by biochemical tests and confirmation was done by the use of API 20 E and API 20 NE in accordance with standard procedures. ESBLs screening was carried out using the double disk synergy test. Data were statistically analyzed using MedCalc statistical software (version 17.2).&#x0D; Results: Total viable bacterial counts (TVBCs) ranged from 1.1 × 103 to 7.1 x 105 cfu/ml; total coliform counts (TCC) ranged from 1.2 x 102 to 3.9 x 103 cfu/ml while total faecal counts (TFC) ranged from 0 to 2.9 × 102 cfu/ml. There were statistical differences in mean TVBCs of the samples (P &lt; 0.05). The mean TCCs of cabbage, carrot and cucumber showed no statistical significance; green beans, green pea and sweet corn also showed no statistical significance (P &gt; 0.05). One hundred and sixty (166) isolates obtained from the samples were identified as Bacillus cereus, Citrobacter freundii, Escherichia coli, Klebsiella pneumoniae, Morganella morganii, Pseudomonas aeruginosa, Proteus mirabilis, Staphylococcus aureus, Serratia marcesens and Staphylococcus saprophyticus. At least one member of all bacterial species, except S. saprophyticus, produced ESBL. &#x0D; Conclusion: This study revealed that salad vegetables could be a vehicle for the spread of extended-spectrum beta-lactamase-producing bacteria which translates to a threat to public health around the world as salads are loved and consumed by all categories of people globally.

Fresh Garlic Extract has a Synergistic Effect with Antibiotics on ESBLs Producing E. coli Urinary Isolates

Aim: Extended spectrum beta-lactamases (ESBLs)-producing bacteria often exhibit a multidrug- resistant phenotype limiting the therapeutic options available to the clinician. This study was conducted to investigate the antibacterial susceptibility pattern of ESBL producing E. coli isolates and evaluation of the antibacterial activity of Garlic extract against it.&#x0D; Study Design: The study was carried out using antibiotics powder and Garlic extract.&#x0D; Place and Duration of Study: At the microbiology lab, Faculty of Medicine, Minia University between January 2017 to August 2018. &#x0D; Methods: The study was carried on 55 ESBL producing E. coli isolates isolated from patients with urinary tract infections (UTI). Screening and confirmation for ESBL was done according to clinical and laboratory standard institute guidelines (CLSI) guidelines. The antibacterial sensitivity to a panel of antibiotics and Garlic was performed by tube dilution method.&#x0D; Results: The results of antibacterial susceptibility testing of ESBLs-producing E. coli isolates in our study showed a higher degree of resistance to the tested antibiotics. In the present study, Garlic showed an inhibitory effect on ESBLs- producing E. coli with concentrations ranging from 10-50 mg/ml and a mean of 30 mg/ml. Garlic extract produce a synergistic effect with all tested antibiotics against all tested ESBL producing E. coli isolates. A Significant decrease in ESBLs genes expression (SHV, TEM and CTX-m) was reported after treatment with Garlic extract (P value: ˂0.001).&#x0D; Conclusion: The combinations of antibiotics and Garlic may be more useful than individual agents, so we recommended that further researches should be undertaken to evaluate the combination of Garlic with antibiotics towards ESBL producing E. coli.

Extended-spectrum beta-lactamase production and multi-drug resistance among Enterobacteriaceae isolated in Addis Ababa, Ethiopia

BackgroundThe global emergence and spread of extended-spectrum beta-lactamases (ESBLs) producing Enterobacteriaceae have been threatening the ability to treat an infection. Hence, this study aimed to determine the prevalence of ESBL-producing and multi-drug resistance (MDR) Enterobacteriaceae (ESBLs-E) from different clinical specimens in Addis Ababa, Ethiopia.MethodsA cross-sectional study was conducted from January 1 to May 30, 2017. A total of 426 Enterobacteriaceae isolates were identified from clinical specimens. The isolates were collected from four laboratories. Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method on Muller Hinton agar (MHA). All Enterobacteriaceae were screened for ESBLs production using cefotaxime and ceftazidime as per CLSI guideline. Each ESBL screening positive Enterobacteriaceae were confirmed by a combination disk test (CDT). Data were entered and analyzed by using SPSS version-20.ResultThe most frequent Enterobacteriaceae were E. coli 228 (53.5%) and K. pneumoniae 103 (24.1%). The magnitude of ESBLs-E was 57.7% (246/426). The highest frequencies of ESBLs-E were observed in blood specimesns (84.4%) and the highest ESBLs production was observed in K. pneumoniae (85.4%). The highest resistance level was seen to sulfamethoxazole-trimethoprim (77.0%), amoxicillin with clavulanic acid (71.6%), cefotaxime (62.2%), cefepime (60.3%) and ceftazidime (60.8%). The overall magnitude of multi-drug resistance (MDR) level was 68.3%. Of ESBLs-E, 96.3% of them were MDR (P < 0.001).ConclusionThere was a high prevalence of ESBLs-E and MDR isolate in Addis Ababa. Most of ESBLs-E was isolated primarily in blood and urine. The highest ESBLs production was observed among K. pneumoniae. Hence, strong infection control strategies must be implemented in hospital settings of the country.

Characterisation of the Resistance Patterns to Non Beta-Lactam Antimicrobials in Esbl-Producing Enterobacteriaceae Isolated from Dogs and Their Owners

Enterobacteriaceae producing extended-spectrum beta-lactamase (ESBL) enzymes are resistant to beta-lactam agents and are also commonly multidrug resistant being associated with the resistance to other classes of antibiotics.The aim of our study was to characterise resistance patterns in non-beta-lactam antibiotics of ESBL-producing Enterobacteriaceae strains isolated from faecal matter of pets and owners.The study was carried out on 63 samples of faecal matter (42 from pets and 21 from owners). The ESBL screening was carried out using the Brilliance ESBL Oxoid chromogenic medium. The isolated strains that generated characteristic presumptive ESBL-producing colonies were cultivated on 5% sheep blood medium for the extraction of bacterial DNA using the boiled preps technique. The confirmation of E. coli species was performed molecularly based on the detection of blauidA and blauspA genes. Other Enterobacteriaceae species were identified based on the minimum biochemical characteristics using the MIU and TSI medium. The phenotypical confirmation of presumptive ESBL-producing strains was carried out using the Double Disc Synergy Test (DDST) using a combination of 3rd generation cephalosporins and beta-lactamase inhibitor agents. The determination of the resistance degree in other classes of antibiotics was carried out through the Kirby-Bauer diffusimetric method, and the results were interpreted according to the CLSI standard.Following the species investigation of isolates, 60/63 (95.28%) belonged to the E. coli species and 3/63 (4.72%) to the K. pneumoniae species. Animal isolates were resistant to sulphonamides (54.76% resistance to SXT), fluoroquinolones (45.23% resistance to ENR) and tetracyclines (54.75% resistance to TE). In addition to strains of animal origin for isolates of human origin, an increased resistance has been noticed to phenicols and aminoglycosides.This study has identified a high prevalence of ESBL-producing Enterobacteriaceae strains and associated with multidrug resistance for pets and their owners.

Screening for ESBL producing bacterial isolates of agricultural soil and profiling for multidrug resistance

Abstract Objectives construction of agricultural environmental soil antibiotic resistance profile of the culturable bacteria in Kalyani, India, followed by the screening of ESBL producers harboring blatem, blashv, and blactx-M. Methods 158 Multi-Drug Resistant (MDR) bacteria were isolated from 18 agricultural sampling points in Kalyani, West Bengal, India followed by their Antibiotic resistance profiling against 15 test antibiotics. They were then screened for the presence of three Extended Spectrum Beta-Lactamase (ESBL) genes, namely blatem, blactx-M and blashv by Double Disk Diffusion test and Polymerase Chain Reaction. The positive candidates were further identified using Amplified Ribosomal DNA Restriction Analysis followed by 16S rDNA sequencing. Results lowest resistance was observed against Vancomycin (0%) whereas the highest against the β lactam Ampicillin (97.5%). Very high resistance was observed against the 3rd generation cephalosporin Cefotaxime (90%). 16/158 bacterial isolates were positive for one or more of the three ESBL genes under investigation. 11 out of those 16 isolates (68.75%) harbored at least 2 genes and the rest 31.25% either had the blashv or the blatem gene. Phylogenetic analysis of the prevalent ESBL carrying organisms in the region demonstrated that the genes are being horizontally transferred across different bacterial orders and classes. Conclusion Significant antimicrobial resistance noted against 12 out of 15 antibiotics tested. Stenotrophomonas, Myroides, Pseudomonas, Enterobacter, and Serratia are found as culturable MDR bacteria carrying ESBL genes among the isolates. It is the first report of environmental ESBL carriage by Myroides isolated from India.

Piperacillin-tazobactam use in ESBL Escherichia coli bacteremia: Should reporting be revised?

Background: While certain extended-spectrum beta-lactamase (ESBL) producing Enterobacteriaceae may appear susceptible to piperacillin-tazobactam in the laboratory, controversy exists regarding its appropriateness to treat serious infections. The Clinical and Laboratory Standards Institute (CLSI) recommends that piperacillin-tazobactam susceptibility be reported as tested, and that ESBL screening is not required in Enterobacteriaceae. We evaluated a sequential group of patients with ESBL Escherichia coli bacteremia to determine the factors associated with piperacillin-tazobactam use despite this controversy. Methods: This was a retrospective observational study at the McGill University Health Center (832 beds) in Montréal, Canada, from April 2010 to June 2015, examining patients with positive blood cultures that grew E. coli. Pediatric, untreated, and duplicate cultures within 14 days were excluded. Antimicrobial susceptibility testing was determined in accordance with CLSI guidelines. Definitive therapy was defined as antibiotics given after susceptibility results became available. Results: There were 845 E. coli bacteremias in the data set, with a median patient age of 68 years (IQR 57–81.25) and a 30-day in-hospital mortality of 12.1%. Overall, there were 44 ceftriaxone-resistant, piperacillin-tazobactam-susceptible bloodstream infections (BSI), and 41 received definitive therapy. Of these, 14/41 (34.1%) received piperacillin-tazobactam and 30 had an infectious disease consult performed. Consult with infectious disease was associated with less use of piperacillin-tazobactam as definitive therapy (7/30 [23%] versus 7/11 [64%]; OR 0.17; 95% CI 0.03–0.96; p = 0.02). Conclusion: Piperacillin-tazobactam definitive therapy was used in approximately one-third of ceftriaxone-resistant E. coli bacteremias. Given a risk of increased mortality with the use of beta-lactam/beta-lactamase inhibitors for treating serious ESBL infections, microbiology laboratories should consider withholding piperacillin-tazobactam susceptibility results in such cases pending definitive evidence of clinical efficacy.