Extended-spectrum Beta-lactamase Screening Research Articles

Prevalence and Molecular Detection of Virulence Genes among Multidrug-Resistant Escherichia coli from Human Clinical Samples and Poultry in Duhok City, Iraq

Abstract Background: Extended-spectrum beta-lactamase (ESBL)-producing, carbapenem-resistant Escherichia coli has increased virulence and multidrug resistance (MDR). Objectives: This study was designed to ascertain the frequency of some virulence factor genes, antibiotic susceptibility patterns, ESBLs, and MDR, focused on colistin-resistant E. coli strains of human and animal origin in Duhok city, Iraq. Materials and Methods: Between December 2020 and April 2021, a total of 150 E. coli isolates (110 from human clinical specimens and 40 from poultry cloacal swabs) were included in this study. The isolates underwent screening for antibiotic susceptibility, MDR, ESBL, and molecular detection of four virulence genes (fimA, cnfL, crL, and hlyA) was conducted using the polymerase chain reaction. Results: Urine specimens (77.2%) compared to blood, wound, vaginal swab, sputum, and semen from outpatients (71.8%). All strains from humans and poultry showed high resistance to ampicillin (86%–97%), ceftriaxone (74%–47%), tetracycline (72%–85%), ciprofloxacin (48%–97%), and colistin (17%–12%). The lowest levels of resistance were found for carbapenems (4%–4%), and the MDR for the isolates was 63%–93%. Apart from carbapenems, colistin-resistant isolates, especially those from poultry, exhibited significant resistance to other antibiotics, and 57% of these isolates being ESBL producers. Three virulence genes (fimA, cnfL, and crL) were highly prevalent (92%) in human isolates, with the crL gene being predominant (100%). Among poultry isolates, fimA was more prevalent (94%) while crL was less common (6%). Conclusion: The predominance of isolates of colistin-resistant poultry origin and the virulence of isolates of human E. coli origin indicate that both strains are currently experiencing an increase in antibiotic resistance.

Comprehensive Profiling of Klebsiella in Surface Waters from Northern Portugal: Understanding Patterns in Prevalence, Antibiotic Resistance, and Biofilm Formation

This study investigates the prevalence of resistance and virulence genes in Klebsiella isolates from surface waters in Northern Portugal, within the broader context of freshwater quality challenges in Southern Europe. The aim of this research is to explain how Klebsiella dynamics, antibiotic resistance, and biofilm formation interact in surface waters. Antimicrobial susceptibility was examined using the Kirby–Bauer disk diffusion method against 11 antibiotics and screening for Extended-Spectrum Beta-Lactamase (ESBL) production using the double-disk synergy. PCR was employed to detect resistance and virulence genes, while biofilm production was assessed using the microplate method. Out of 77 water isolates, 33 Klebsiella (14 Klebsiella spp. and 19 K. pneumoniae strains) were isolated. ESBL production was observed in 36.8% of K. pneumoniae and 28.6% of Klebsiella spp. High resistance rates to blaCTX-U were observed in both. The papC gene was prevalent, signifying potential environmental risks. Biofilm production averaged 81.3% for K. pneumoniae and 86.9% for Klebsiella spp. These findings underscore the intricate interplay between Klebsiella’s dynamics and freshwater quality, with ESBL’s prevalence raising concerns about waterborne dissemination and public health implications. This work supports the need for vigilance of Klebsiella in surface waters in Southern Europe.

Extended-spectrum beta-lactamases in clinical isolates of Escherichia coli and Klebsiella pneumoniae recovered from patients at the Tamale Teaching Hospital, Ghana.

Extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli and Klebsiella pneumoniae are pathogens of significant public health interest for which new antibiotics are urgently needed. To determine the prevalence of ESBLs in E. coli and K. pneumoniae isolates from patients attending the Tamale Teaching Hospital (TTH) in Ghana. The study was a cross-sectional study involving convenience sampling of E. coli and K. pneumoniae isolates from consenting patients' clinical specimens, between April and June 2015. Antimicrobial susceptibility test was performed, and ESBL-producer phenotypes were further screened for BlaTEM, BlaSHV, and BlaCTX-M genes. Patients' clinical data were additionally collected using a structured questionnaire. Of the 150 non-duplicate E. coli and K. pneumoniae isolates identified, 140 were confirmed as E. coli (84%, n = 117) and K. pneumoniae (16%, n = 23). Of these, sixty-two (44%) [E. coli (84%; n = 52); K. pneumoniae (16%; n = 10)] phenotypically expressed ESBLs. The proportion of ESBL-producing isolates was higher in adults (15-65 years) than in neonates (< 28 days) (p = 0.14). Most of the isolates showed a high percentage resistance to ampicillin (96%) and tetracycline (89%), but a relatively lower resistance to amikacin (36%). No isolate was resistant to meropenem. More ESBL producers were multidrug resistant compared to non-ESBL-producers [23% (14/62) versus 18% (14/78); p = 0.573]. Overall, 74% (n = 46) of the ESBL genotypes expressed BlaCTX-M-1 genes, followed by 63% (n = 39) BlaTEM, and 16% (n = 10) BlaSHV. The study showed a high prevalence of ESBL-positive E. coli and K. pneumoniae, mostly CTX-M-1 producers at TTH. Routine laboratory ESBL screening is warranted to inform patient management.

PREVALENCE AND CHARACTERISATION OF EXTENDED-SPECTRUM BETA-LACTAMASES AND PLASMID-MEDIATED QUINOLONES RESISTANCE IN Enterobacteriaceae ISOLATED FROM COMPANION ANIMALS

Antimicrobial resistance is a major public health concern worldwide. This study aims to determine the prevalence of Enterobacterales producing beta-lactamase (TEM, SHV, OXA) or extended-spectrum beta-lactamases (ESBL), as well as plasmid-mediated resistance to quinolones (PMQR) (qnrA, qnrB, qnrS) in companion animals from the northeast region of Romania. A total of 124 faecal samples were collected aseptically from healthy dogs attending the veterinary practice for vaccination and cultivated on Brilliance ESBL medium (Oxoid, UK). The ESBL production testing was performed using the combination disc test. The identification of Enterobacterales strains was achieved using molecular identification and based on biochemical tests. Antimicrobial susceptibility testing was performed using the disk diffusion method. Identification of genes encoding for beta-lactamase enzymes and genes encoding plasmid-mediated resistance to quinolones was performed by PCR according to the protocols previously described. After ESBL screening, 31 (31/124; 25%) extended-spectrum cephalosporin (ESC)-resistant Enterobacterales were obtained, and 67.74% (21/31) of them were confirmed as ESBL-producers. Regarding the Enterobacterales species, 27 (27/31; 87.1%) were Escherichia coli and 4 (4/31; 12.9%) strains were Klebsiella pneumoniae. Among the ESBL-producing isolates, the blaCTX-M-1 gene group was predominant (58.82%), followed by the blaCTX-M-9 group (41.18%). The blaTEM, blaSHV and blaOXA gene groups were identified in 54.83%, 29.03% and 3.22% of the analysed strains, respectively. The prevalence of PMQR genes was 22.58% and consisted only of qnrS (19.35%) and qnrA (3.22%) genes. The prevalence of ESBL strains related to the total number of analysed samples was 16.93% (21/124). The findings show a significant prevalence of ESBLs and PMQR genes in Enterobacterales strains isolated from the faeces of healthy dogs, implying that pets may pose a risk of transmitting ESBL strains to other animals or owners.

Emerging Trends of Extended Spectrum BetaLactamase and Metallo-Beta Lactamase Escherichia Coli Strains among Pregnant Women in Mile 4 Hospital, Abakaliki: Implications for Antenatal Care

This investigation assessed the prevalence of extended spectrum beta-lactamase (ESBL) and metallo-beta lactamase (MBL) producing Escherichia coli among seemingly healthy pregnant women attending the antenatal clinic at Mile 4 Hospital in Abakaliki, Nigeria. A total of 100 midstream urine samples were collected and processed using standard microbiology techniques. Antibiotic susceptibility testing was conducted via the disc diffusion method. ESBL and MBL production screening utilized the double disc synergy test and the combined disc test inhibition assay, respectively. The findings demonstrated a 54% prevalence of E. coli among the study participants, with the highest occurrence observed in the third trimester (31.5%) and the lowest in the first trimester (25.9%). Across age groups, prevalence ranged from 9.3% in participants aged ≤20 years to 44.4% in the 21-30 age bracket. Additionally, among participants with varying educational backgrounds, prevalence ranged from 18.5% in uneducated individuals to 33.3% in those with a secondary education. Antibiotic susceptibility of the isolates varied from 0% to 81.5%. ESBL and MBL screening identified 18 (33.3%) and 11 (20.4%) positive isolates, respectively. The antibiotic susceptibility among ESBL-producing E. coli ranged from 0% to 100%, while MBL-positive E. coli showed susceptibility ranging from 0% to 90.5%. The multiple antibiotic resistance index (MARI) ranged from 0.3 to 1.0, with an average of 0.7 among the isolates. This study highlights a substantial prevalence of multidrug-resistant ESBL and MBL-producing E. coli strains among apparently healthy pregnant women. This situation raises concerns about the early exposure of fetuses to multidrug resistance, emphasizing the urgency for proactive measures to address this emerging health risk. Keywords: ESBL positive E. coli, metallo-betalactamase positive E. coli, apparently healthy pregnant women, Abakaliki, Nigeria.

The notable relatedness between ESBL producing Enterobacteriaceae isolated from clinical samples and asymptomatic fecal carriers

BackgroundThe investigation of the presence of extended-spectrum beta-lactamase (ESBL) within Enterobacteriaceae in both fecal carriers and patients is an essential matter. Furthermore, the assessment of distinct characteristics exhibited by resistant bacteria obtained from fecal carriers and patients, as well as the comparison of these characteristics between the two groups, could provide a deeper understanding of how the resistant isolates can remain concealed within a dormant reservoir and intensify antimicrobial resistance. The aim of the present study was to concentrate on the comparison of the antimicrobial resistance pattern and molecular features between strains obtained from clinical and carrier sources.Material and methodsA total of 142 clinical samples and 120 rectal swabs were collected from June to October 2016. ESBL screening was performed using the double-disk synergy test. PCR was done for the detection of ESBL genes. Assessment of biofilm formation, virulence factor genes, and MLVA was performed for K. pneumonae isolates. Phylogroup typing was performed for E. coli isolates.ResultsOf 146 samples, 67.6% were E. coli, and 32.4% were K. pneumoniae. The rate of blaCTXM-15 was 89.4%. In K. pneumoniae type D, ompk35 and fimH were the highest. All the K. pneumoniae isolates were classified into 12 mini clusters and the clinical isolates were characterized into 7 mini clusters. The phylogroup B2 in ESBL-EC was the highest (56.2%).DiscussionComparison of molecular characteristics and clonal relatedness between fecal carriers and patients showed noticeable relatedness and similarity which may indicate that ESBL-KP can be colonized with the same profiles in different settings and, therefore, may be widely distributed in both community and hospital settings. Therefore, implementation of control protocols, including surveillance of the fecal carriers, could impressively reduce silent reservoirs without clinical symptoms as well as patient rates.

Abundance and prevalence of ESBL coding genes in patients undergoing first line eradication therapy for Helicobacter pylori.

The spread of extended-spectrum beta-lactamases (ESBLs) in nosocomial and community-acquired enterobacteria is an important challenge for clinicians due to the limited therapeutic options for infections that are caused by these organisms. Here, we developed a panel of ESBL coding genes, evaluated the abundance and prevalence of ESBL encoding genes in patients undergoing H. pylori eradication therapy, and summarized the effects of eradication therapy on functional profiles of the gut microbiome. To assess the repertoire of known beta lactamase (BL) genes, they were divided into clusters according to their evolutionary relation. Primers were designed for amplification of cluster marker regions, and the efficiency of this amplification panel was assessed in 120 fecal samples acquired from 60 patients undergoing H. pylori eradication therapy. In addition, fecal samples from an additional 30 patients were used to validate the detection efficiency of the developed ESBL panel. The presence for majority of targeted clusters was confirmed by NGS of amplification products. Metagenomic sequencing revealed that the abundance of ESBL genes within the pool of microorganisms was very low. The global relative abundances of the ESBL-coding gene clusters did not differ significantly among treatment states. However, at the level of each cluster, classical ESBL producers such as Klebsiella sp. for blaOXY (p = 0.0076), Acinetobacter sp. for blaADC (p = 0.02297) and others, differed significantly with a tendency to decrease compared to the pre- and post-eradication states. Only 13 clusters were common across all three datasets, suggesting a patient-specific distribution profile of ESBL-coding genes. The number of AMR genes detected in the post-eradication state was higher than that in the pre-eradication state, which could be attributed, at least in part, to the therapy. This study demonstrated that the ESBL screening panel was effective in targeting ESBL-coding gene clusters from bacterial DNA and that minor differences exist in the abundance and prevalence of ESBL-coding gene levels before and after eradication therapy.

Molecular Detection of Extended Spectrum Beta-Lactamase Gene in Clinical Isolates of Klebsiella Species in Jos North Central of Nigeria

Aims: The aim of the study was to determine the antimicrobial susceptibility of Klebsiella species, prevalence of Extended Spectrum Beta-Lactamases (ESBLs) and the gene coding ESBL production in Klebsiella Species obtained from clinical specimens of patients attending Plateau State Specialist and Jos University Teaching Hospitals both in Jos North Central of Nigeria.&#x0D; Study Design: This was hospital based study conducted in the Department of Medical Microbiology unit of Plateau State Specialist and Jos University Teaching Hospital (JUTH) during the periods of January to April 2020.&#x0D; Methodology: Out of 156 non-motile, oxidase negative gram negative organisms collected, 137 isolates were identified to be Klebsiella species using oxoid Microbact 24E kit for Enterobacteriaceae identification and Microsoft for bacteria identification. Antimicrobial Susceptibility Testing (AST) and ESBL screening was done by Kirby-Bauer disk diffusion method using oxoid Single disk antibiotics which include Ceftazidime (30µg), Cefotaxime (30µg), Imipenem (10µg), Ciprofloxacin (5µg), Gentamicin (10µg), Ampicillin (10 µg), Aztreonam (30 µg), and Azithromycin (15 µg) on Mueller Hinton Agar (oxoid).Break point was determined by CLSI AST guidelines. The isolates that showed resistance or reduced susceptibility to Ceftazidime and Cefotaxime are suspected to be ESBL positive Klebsiella. Phenotypic confirmation of ESBL positive isolates was carried out using Double Disk Synergy Test (DDST) with Ceftazidime 30µg (oxoid), Cefotaxime 30µg (oxoid) and Amoxicillin Clavulanic acid 20/10ug (Oxoid).Genotypic detection of ESBL gene was done using Polymerase Chain Reaction (PCR).&#x0D; Results: Antimicrobial Susceptibility Test showed Imipenem to be highly sensitive to ESBL positive Klebsiella pneumonia with 100% sensitivity. Gentamicin, Ampicillin, and Azithromycin showed reduced susceptibility. Genotypic results showed presence of bla TEM, bla SHV genes at 868bp and 972bp band size with no occurrence of bla CTX-M gene. There was multiple occurrence of gene in some of the isolates tested while some recorded no ESBL gene. The ESBL prevalence of 15.3% was recorded in this study.&#x0D; Conclusions: The prevalence of 15.3% recorded in this study is low compared to other studies from some other states within the country. Efforts should be put in place to ensure non-proliferation of this ESBL Klebsiella strain as it creates not only a therapeutic challenge but health problem.

Analysis of Extended Spectrum Beta Lactamase Frequency in Klebsiella spp Isolates

The issue of increasing resistance to antibiotics in recent years has become an important problem all over the world. Our aim is to determine the antimicrobial resistance profile and Extended Spectrum Beta-Lactamase (ESBL) rates in Klebsiella spp isolates to prevent the gradual increase in multi-resistant isolates as a result of unconscious antibiotic use thereby contributing to the faster effective treatment of infections. A total of 100 Klebsiella spp were isolated and identified from various clinical specimens. Antibiotic susceptibility tests were performed using the Kirby-Bauer method. The presence of extended-spectrum beta-lactamases (ESBL) was detected using the Double Disc Synergy Test (DDST) and E-test methods. The rates of ESBL-producing strains were 46.1% in 6 K. oxytoca and 56.3% in 49 K. pneumoniae. These strains were found to be 38% in 38 adult patients and 17% in 17 pediatric patients, and this difference was statistically significant (p &amp;lt;0.05). The ESBL rate was 31% in 31 male patients and 24% in 24 female patients, and this difference was not statistically significant (p&amp;gt;0.05). This rate was found to be high in patients hospitalized in the pediatric service and intensive care unit. 67 out of 100 strains were found to be suspicious for ESBL by Disk Diffusion Test (DDT). DDST and E-tests were applied as confirmatory tests. The sensitivity of the DDST and E tests was 100%. Screening for ESBL in Klebsiella spp and other members of Enterobacteriaceae isolates is necessary to reduce further selection and spread of these increasingly broad-spectrum antimicrobial-resistant enteric pathogens.

Comparative Analysis between Salmonella enterica Isolated from Imported and Chinese Native Chicken Breeds.

Salmonella enterica is considered a significant threat to the global poultry industry and public health. In recent decades, antimicrobial resistance in Salmonella enterica has attracted increasing concern throughout the world. However, limited information is available on Salmonella enterica among different breeds of breeder chickens. Thus, this study aimed to compare the prevalence, serotype distribution, emergence of extended-spectrum beta-lactamases (ESBLs), antimicrobial resistance, and genetic resistance mechanisms in Salmonella enterica among different breeds of breeder chickens. A total of 693 samples (dead embryos, cloacal swabs, water, feed, environmental swabs, and meconium of newly hatched chicks) were selected and cultured for Salmonella from four breeder chicken farms in Shandong province, China, representing one imported and three native breeds, and the isolates were further serotyped. Of the Salmonella isolates, susceptibility to 11 antimicrobials of 5 classes, ESBL screening, and the presence of 21 antimicrobial resistance genes were determined in the present study. Overall, 94 (13.6%) isolates were recovered, which were divided into 3 serotypes (Salmonella Pullorum (n = 36), Salmonella Thompson (n = 32), and Salmonella Enteritidis (n = 26)). The results showed that the prevalence of Salmonella enterica isolates from the imported breeds was higher compared with the three domestic breeds. Eight of the ninety-four isolates were ESBL-positive strains, which were recovered from a domestic breed chicken farm. These eight ESBL-producing isolates were serotyped to Pullorum. Surprisingly, Salmonella Enteritidis (S. enteritidis) and S. pullorum were simultaneously isolated from a single dead embryo observed among one native breed. Meanwhile, among the Salmonella isolates, 53.2% (50/94) were multidrug-resistant strains, and 44.7% (42/94) of the isolates presented resistance to at least five antibiotics. Nearly all of the isolates (97.9%, 92/94) were resistant to at least one antimicrobial; one isolate of S. Thompson was resistant to seven antimicrobial agents belonging to four different classes. The carriage rate of three resistance genes (tetA, tetB, and sul1) among isolates from the imported breeds (87%, 70%, and 65.2%) was higher than that in those from domestic breeds (35.2%, 36.6, and 14.1%). To our knowledge, this is the first report of ESBLs-producing Salmonella isolated from a Chinese native breed of breeder chickens. Our results also highlight that a high prevalence of multidrug-resistant Salmonella enterica contamination is widespread among different breeds of breeder chickens, which is a major risk of food-borne diseases and public health.

Prevalence, Characterization, and Antimicrobial Resistance of Extended-Spectrum Beta-Lactamase-Producing Escherichia coli from Domestic Free-Range Poultry in Agogo, Ghana

Poultry has been suggested as an important source for extended-spectrum beta-lactamase (ESBL)-producing bacteria that can lead to difficult-to treat infections in humans. Therefore, this study aims to determine the frequency, the genetics, and antimicrobial resistance profiles of ESBL-producing Escherichia coli in domestic free-range poultry in Agogo, Ghana. The study was set up and piloted from January 2019 until June 2019. Between June and December 2019, fecal samples (N = 144) were collected from free-roaming chickens from domestic farms in the regions of Sukuumu, Bontodiase, and Freetown and cultured on ESBL screening agar. Strain identification and antibiotic susceptibility were performed using the VITEK 2 compact system. ESBL-producing E. coli were confirmed using the double disk synergy test. Molecular characterization of ESBL-associated genes (blaTEM, blaSHV, and blaCTX-M) were performed using conventional polymerase chain reaction (PCR) and further sequencing of obtained PCR amplicons. The result showed that 56.2% (n/N = 81/144) of collected fecal samples were positive for ESBL-producing E. coli. Majority of the isolates showed resistance to tetracycline (93.8%, n/N = 76/81) and trimethoprim-sulfamethoxazole (66.7, n/N = 54/81), whereas resistance to carbapenems was not found. The majority of ESBL-producing E. coli carried the blaCTX-M genes, with blaCTX-M-15 being the dominant (95.1%, n/N = 77/81) genotype. In this study, we report high frequencies of ESBL-producing E. coli in smallholder free-range poultry representing a potential source of infection, highlighting the need for control of antibiotic use and animal hygiene/sanitation measures, both important from a One Health perspective.

Bacterial profile and extended spectrum beta lactamase screening of urinary tract infection among asymptomatic and symptomatic pregnant women attending antenatal care in ALERT Hospital, Addis Ababa, Ethiopia.

The occurrence of extended spectrum beta lactamase-producing uropathogens, especially in pregnant women can result in life-threatening condition and morbidity for both the mother and the newborn due to very limited drug options for treatment of these pathogens. The aim of this study was to determine the bacterial profile, associated factors, and their antimicrobial susceptibility patterns and to identify extended spectrum beta lactamase-producing bacterial uropathogens. A hospital-based cross-sectional study was conducted from July to September 2018 on a total of 177 pregnant women with and without symptoms of urinary tract infection at ALERT Hospital, Addis Ababa, Ethiopia. From these study participants, 72 have symptoms, whereas 105 have no symptoms. All urine samples were inoculated onto cysteine lactose electrolyte deficient medium and MacConkey agar. Colonies were counted to check the presence of significant bacteriuria. Pure isolates of bacterial pathogen were characterized and identified at species level by colony morphology, gram stain, and standard biochemical procedures. All Gram-negative isolates were put into Muller-Hinton agar plates for antibiotic susceptibility test by Kirby-Bauer disc diffusion technique. Extended spectrum beta lactamase was detected using double-disk synergy methods on Muller-Hinton agar. The data were double entered into epidemiological Information system and analyzed using Statistical Package for Social Science version 26. The overall proportion of urinary tract infection among pregnant women was 14.7% (n = 26/177). Klebsiella pneumoniae was the predominant bacterial etiologic agent of urinary tract infection 26.9% (n = 7/26). The proportion of extended spectrum beta lactamase among Gram-negative isolates was 50% (n = 6/12). Among extended spectrum beta lactamase-producing isolates (100%), all are resistance to amikacin and gentamicin while intermediate level resistance rate of 66.7% was observed among trimethoprim-sulphamethoxazole. They were susceptible for some limited drugs, and these were Nitrofurantoin (83.3%) and Chloramphenicol (83.3%). Majority of extended spectrum beta lactamase-producing isolates exhibited co-resistance to other commonly prescribed antibiotics. This indicates that the option of treatment for these pathogens rapidly decreased from time to time which results serious life-threatening conditions, especially in mother and newborn unless the appropriate measure is taken.

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

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

Cefixime and cefixime-clavulanate for screening and confirmation of extended-spectrum beta-lactamases in Escherichia coli

IntroductionDetection of Extended-Spectrum Beta-Lactamases (ESBLs) depends on screening for resistance to certain cephalosporins, confirmation with selective ESBL inhibitors, and ESBL genes detection. New tests are required for accurate ESBL detection.AimsTo test the ability of cefixime (CFM) and cefixime-amoxicillin/clavulanate (CFM-AMC) as a screening and confirmatory test for ESBL identification.Methods246 clinical isolates of Escherichia coli were tested by an ESBL screening test, a double-disk synergy test (DDST), a disk replacement test, the Vitek 2 ESBL test, and an ESBL genes test by PCR. CFM ESBL Screening was performed by disk diffusion, while CFM-AMC confirmation was performed by DDST and a disk replacement test.Results246 E. coli clinical isolates from two referral hospitals were collected over 2 years. The mean age ± standard deviation of patients was 43.8 ± 27.7 years and 76.8% were females. Resistance rates to penicillins, first, second, and third generation cephalosporins, and monobactams were very high at 97%, 84%, 100% and 97%, respectively. ESBL screening was positive in 81.3% of isolates, DDST was positive in 74.8%, disk replacement was positive in 79%, Vitek 2 ESBL test was positive in 67.3%, and ESBL genes were detected in 85.8% of isolates (CTX-M 75%, TEM 42.5%, SHV 4.6%). Compared to genotyping, screening with CFM achieved 87.7% sensitivity and 64.7% specificity. CFM-AMC DDST achieved 75.8% sensitivity and 75.4% specificity, and CFM-AMC disk replacement had 73% sensitivity and 70% specificity.ConclusionsHigh prevalence of ESBLs was noted among E. coli isolates, dominated by CTX-M genotype. ESBL screening and confirmation using CFM and CFM-AMC is a new and accurate method for ESBLs detection.

High Magnitude of Fecal Carriage of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae at Debre Berhan Comprehensive Specialized Hospital, Ethiopia.

BackgroundGastrointestinal colonization rate of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) is the major risk factor for infection and dissemination of resistance clones in healthcare facilities. This study aimed to investigate the magnitude of the fecal carriage of ESBL-PE and associated factors among hospitalized patients at Debre Berhan Comprehensive Specialized Hospital, North Shoa, Amhara Regional State, Ethiopia.MethodsA hospital-based cross-sectional study was conducted among 383 hospitalized patients from November 2020 to March 2021. Stool sample or rectal swab was aseptically collected and cultured on different culture media for isolation of Enterobacteriaceae. Identification was done by conventional biochemical tests. Screening of extended-spectrum beta-lactamase (ESBL) production was done by using cefotaxime and ceftazidime and confirmed by the combination disk method. Data analysis was performed by Statistical Package for Social Sciences software version 25 and a P-value ≤0.05 was considered as statistically significant.ResultsFrom the total of 383 hospitalized patients, a total of 347 Enterobacteriaceae were isolated. The overall gastrointestinal colonization rate of ESBL-PE was 47.3% (164/347). The predominant ESBL-PE were E. coli 54.9% (90/164) and K. pneumoniae 33.5% (55/164). The overall multi-drug resistance rate (MDR) was 87.8% (305/347). The highest resistance was observed to ampicillin (98.3%), followed by gentamicin (80.7%), and tetracycline (73.3%), respectively. ESBL-PE were highly susceptible to meropenem (90.2%) and imipenem (89.0%). History of antibiotic use in the past 3 months (p<0.001), admission in the neonatal intensive care unit (p=0.023), and presence of chronic disease (p<0.001) were independently associated with fecal carriage of ESBL-PE.ConclusionThe magnitude of ESBL-PE and MDR was high in the study area. Meropenem and imipenem were active against ESBL-PE. Therefore, strict infection control measure is needed in the study area to limit the infection and dissemination of ESBL-PE.

Assessment of extended – spectrum beta lactamases (ESBL) gene among gram-negative bacilli bacteria causing community acquired urinary tract infection among females in southwestern Nigeria

Background: Urinary tract infections (UTI) is commoner in females and mainly caused by Gram Negative bacilli. The emergence of Extended – spectrum beta lactamases (ESBL) has made its treatment difficult especially in the community because of limited therapeutic options. This study was undertaken to determine the prevalence of ESBL-producing Gram negative bacilli causing UTI among females in Babcock University Community; evaluate the associated risk factors and to determine the prevalence of the associated genes among the ESBL isolates.&#x0D; Methods: A descriptive cross-sectional study of which 200 female participants with clinical UTI were recruited in Babcock University Community. Pretested interviewer–based structured questionnaire was used to obtain the socio-demographic and risk factors. Mid-stream urine was also collected for evaluation of ESBL- producing Gram-negative bacilli. DNA extraction was done for ESBL positive isolates and SHV, TEM, CTX-M and OXA ESBL genes were detected by PCR. SPSS Version 23.0 was used for data analysis.&#x0D; Results: The 200-urine samples tested from the participants yielded 33-Gram negative bacilli isolates. Out of these 33 isolates, 17/33(51.5%) were ESBL-producers giving prevalence rate of 8.5% among the participants. The distribution of the ESBL genes among the ESBL isolates was as follows: SHV 12/17(70.6%), TEM 4/17(23.5%), CTX-M 3/17(17.6%) and OXA 5/17(29.4%). No associated risk factors were found.&#x0D; Conclusion: All the four types of ESBL genes evaluated were present among the participants. Routine screening for ESBL for Community acquired UTI is necessary to curb treatment failure.

Assessment of extended – spectrum beta lactamases (ESBL) gene among gram-negative bacilli bacteria causing community acquired urinary tract infection among females in southwestern Nigeria

Background: Urinary tract infections (UTI) is commoner in females and mainly caused by Gram Negative bacilli. The emergence of Extended – spectrum beta lactamases (ESBL) has made its treatment difficult especially in the community because of limited therapeutic options. This study was undertaken to determine the prevalence of ESBL-producing Gram negative bacilli causing UTI among females in Babcock University Community; evaluate the associated risk factors and to determine the prevalence of the associated genes among the ESBL isolates.&#x0D; Methods: A descriptive cross-sectional study of which 200 female participants with clinical UTI were recruited in Babcock University Community. Pretested interviewer–based structured questionnaire was used to obtain the socio-demographic and risk factors. Mid-stream urine was also collected for evaluation of ESBL- producing Gram-negative bacilli. DNA extraction was done for ESBL positive isolates and SHV, TEM, CTX-M and OXA ESBL genes were detected by PCR. SPSS Version 23.0 was used for data analysis.&#x0D; Results: The 200-urine samples tested from the participants yielded 33-Gram negative bacilli isolates. Out of these 33 isolates, 17/33(51.5%) were ESBL-producers giving prevalence rate of 8.5% among the participants. The distribution of the ESBL genes among the ESBL isolates was as follows: SHV 12/17(70.6%), TEM 4/17(23.5%), CTX-M 3/17(17.6%) and OXA 5/17(29.4%). No associated risk factors were found.&#x0D; Conclusion: All the four types of ESBL genes evaluated were present among the participants. Routine screening for ESBL for Community acquired UTI is necessary to curb treatment failure.

Early Detection of Drug Resistant Enterobacteriaceae in Urinary Tract Infections using Chromogenic Agar Medium in a Tertiary Care Hospital, Kakinada, Andhra Pradesh, India

Introduction: The irrational and inappropriate use of beta lactam antimicrobial drugs has led to the advent of Extended Spectrum Beta-Lactamase (ESBL) resistant strains. ESBL producing Enterobacteriaceae strains are frequent causative agents both in community and in acquired nosocomial infections and Urinary Tract Infections (UTI). The phenotypic confirmatory tests rarely identify all ESBLs. Chrom ID (Chromogenic identification Media) ESBL – Bx (bioMerieux) is a completely new and innovative chromogenic medium designed specifically for the screening of ESBL producing Enterobacteria directly from urine samples. It is a ready to use selective media which is sensitive and specific for rapid and presumptive identification of ESBL producing Enterobacteriaceae. Aim: Early detection of ESBL producing Enterobacteriaceae directly from urine samples on chromogenic medium (Chrom ID- ESBL- Bx) and confirmation of ESBL producing Enterobacteria using Disc Potentiation Test (DPT). Materials and Methods: The present cross-sectional study was conducted in the Department of Microbiology, Rangaraya Medical College, Kakinada, Andhra Pradesh, India from November 2019 to March 2020 (five months duration). The study was done on 70 urine samples from patients with UTI. All samples were subjected to wet mount, inoculated directly for culture on Chrom ID ESBL- Bx agar and MacConkey agar. Antibiotic Susceptibility testing of ceftazidime and cefotaxime was done by Kirby-Bauer disc diffusion method and conformation of ESBL production by DPT using Clinical and Laboratory Standards Institute (CLSI) method. The Statistical Package for the Social Sciences (SPSS) Statistical package version (18.0) was used. Results: A total of 56 (80%) isolates were obtained from 70 urine samples, out of them 28 (50%) were Escherichia coli, 21 (37.5%) were Klebsiella spp., 7 (12.5%) were Proteus spp., 23 (82.14%) isolates of Escherichia coli, 15 (71.43%) of Klebsiella spp., 6 (85.71%) of Proteus spp., isolated were screened positive using Chrom ID ESBL-Bx agar. About 44 (78.57%) of total Enterobacteria (56) were screened for ESBL production. 20 (86.96%) of Escherichia coli, 11 (73.33%) of Klebsiella spp., and 5 (83.33%) of Proteus spp., that were screened positive using Chrom ID ESBL agar were confirmed (by DPT) as ESBL producers and 2 (16.6%) of total (12) isolates that were screened negative by Chrom ID ESBL agar were confirmed as ESBL producers when screened and confirmed by DPT. So sensitivity and specificity CHRO Magar was 94.73% and 55.5%. Conclusion: ESBL continues to become a serious public health threat. Results from present study showed that CHROMagar ESBL has a high sensitivity and a convenient method for making provisional diagnosis of drug resistant Enterobacterial infections in 24 hours. Chrom ID ESBL- Bx agar medium allows easy differentiation of different bacteria based on colony colouration.

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 &lt; 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&lt;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.