Spectrum Beta-lactamase Production Research Articles

Extended spectrum and metalo beta lactamase producing gram negative bacterial pathogens from cockroaches collected at hospital, Southern Ethiopia

BackgroundCockroaches can pose a significant health risk in hospital environments because they may serve as reservoirs and vectors for nosocomial pathogens. Cockroaches harbor epidemiologically significant extended spectrum and metalo beta lactamase producing Gram negative bacterial pathogens, which complicate nosocomial infections.ObjectivesThe main aim of this study is to determine aetiology and phenotypic extended spectrum and metalo beta lactamase producing Gram negative bacteria pathogens from cockroaches collected in hospitals.MethodsA cross-sectional study was employed from February to May 2022 to determine the antibiotic resistance producing bacterial isolates from cockroaches by giving special emphasis to metalo beta lactamase and extended spectrum beta lactamase production from different wards of WSUCSH. Cockroaches were collected with hands wearing sterile gloves. External homogenate was prepared and incubated microbiologically by using different culture media and differentiated biochemically. Antimicrobial susceptibility testing was performed by disk diffusion method. ESBL production was conducted using double disc synergy method and double disk method was used to detect MBL enzyme detection. Descriptive statistics was used to determine prevalence and percentage.ResultOut of 245 cockroaches, 108 Gram negative bacteria were isolated. K. pneumoniae 29(26.9%) was the most predominant bacteria and Enetrobacter spp. 8(7.4%), was the least. All, K. pneumoniae, P. mirabilis, and Enterobacter isolates were pan-resistant to Ampicillin. P.aeruginosa and P.mirabilis antibiotics showed ≥ 80% resistant for amoxicillin/clavulanic acid antibiotics. Cefotaxime, ceftazidime, ceftriaxone and imipenem showed relative efficacy compared with other antibiotics. Out of 78 amoxicillin-clavulanic acid resistant isolates, 42(34.7%) were ESBL producers. ESBL production is more depicted by P. aeruginosa, A. baumannii, K. pneumoniae and E. coli. The overall prevalence of MBL production is 29(23.1%). K. pneumoniae P. aeruginosa, E.coli, A. baumannii, Enterobacter spp and K.oxytoca revealed MBL production.ConclusionThe overall prevalence of ESBL and MBL producing nosocomial agents from hospital cockroaches was 34.7% and 23.1% respectively. P.aeruginosa, A.baumannii, K.pneumoniae and E.coli showed pronounced ESBL production. All bacterial isolates except P. mirabilis and C. freundii showed MBL production. The needed to evaluate our antibiotic stewardship program and antibiotic resistance detection for treatment is mandatory. The impact of cockroach as a source of AMR should be sought.

PREVALENCE OF ASYMPTOMATIC BACTERIURIA IN ANTENATAL WOMEN ATTENDING TERTIARY CARE HOSPITAL-A CROSS-SECTIONAL STUDY

Objective: Urinary tract infection (UTI) is one of the most common bacterial infections during pregnancy due to anatomical changes and physiological adaptations during pregnancy. Asymptomatic bacteriuria is the significant presence of bacteria in the urine of an individual without symptoms. Untreated asymptomatic bacteriuria (ASB) in pregnancy predisposes to symptomatic UTI in 25% of infected women. Screening of antenatal women help in early diagnosis and treatment of ASB and thus to prevent maternal and fetal morbidity and mortality. The present study was carried out to determine the prevalence of UTI in pregnant women and to study the bacteriological profile and antimicrobial sensitivity patterns of uropathogens. Methods: A Cross-sectional study was conducted for a period of six months and midstream urine specimens were collected from 480 pregnant females and were processed by standard protocols. All subjects were clinically identified to have no signs and symptoms of UTI. Antibiotic susceptibility testing was done as per CLSI guidelines. Results: Prevalence rate of asymptomatic bacteriuria was seen 10% in pregnant women. Majority of the culture-positive patients belonged to the age group of 26-30 y (31.25%). 70.84% were Gram-negative isolates and 29.16% were Gram-positive organisms. The commonest pathogen isolated was Escherichia coli (33.33%). In the present study, Extended Spectrum Beta-Lactamase (ESBL) production was seen in (20.58%) isolates, and Metallo Beta-Lactamase (MBL) production was seen in (17.64%) isolates. Conclusion: This study reveals the importance of screening of pregnant women for UTI. Emerging multi-drug resistance seen in uropathogens emphasizes the need to rationalize use of antibiotics, which eventually prevent development of resistant strains.

Extended-Spectrum Beta-Lactamase Producing Enterobacterales Infections in the United States, 2012-2021

Background: The 2022 Special Report: COVID-19 U.S. Impact on Antimicrobial Resistance identified continued increases in the rate of extended- spectrum beta-lactamase producing (ESBL) infections in the United States from 2017 through 2020. Using similar data sources and methodology, we examined the trends of species-specific ESBL infections from 2012-2021. Methods: We identified a cohort of patients from the PINC AI and BD Research Insights databases with a clinical culture yielding a Klebsiella pneumoniae or Escherichia coli isolate with accompanying susceptibility testing. E. coli or K. pneumoniae isolates non-susceptible to ceftriaxone, cefotaxime, ceftazidime, or cefepime were considered suggestive of ESBL production. Isolates from patients with no culture yielding the same resistance phenotype of interest in the previous 14 days were counted as an incident case. Community-onset (CO) cultures were obtained ≤ day 3 of hospitalization; hospital-onset (HO) cultures were obtained ≥ day 4. We used a raking procedure to determine weights for extrapolating the number of discharges included in our sample to match the distribution of discharges, stratified by bed size, U.S. census division, urban/rural designation, and teaching status, for U.S. hospitals included in the American Hospital Association survey. We evaluated rates over time due to the changes in number of hospitalizations during the COVID-19 pandemic. Results were stratified by HO and CO, and sterile and non-sterile specimen sources. Results: In 2021, there were 48,936 ESBL K. pneumoniae and 153,112 ESBL E. coli infections among approximately 32 million discharges. Overall, most infections were CO and from non-sterile specimens. From 2012-2021, the rate of ESBL K. pneumoniae increased from 9.54 to 15.28 per 10,000 discharges. ESBL E. coli infections increased from 2012-2020 (30.18 to 51.32 per 10,000 discharges), then declined in 2021 (47.81 per 10,000 discharges) (Table 1, Figure 1). The proportion of non-sterile ESBL E. coli declined from 88% in 2012 to 83% in 2021, and the proportion of non-sterile ESBL K. pneumoniae was 85-87% over the study period (Figure 2). Conclusion: ESBL E. coli and K. pneumoniae infections increased from 2012-2021, although the CO ESBL E. coli rate decreased between 2020 and 2021. Understanding changes in culturing practices over time may provide insights into the increased proportion of ESBL E. coli from sterile sites. Additionally, further investigation into differences in organism trends, particularly in 2021, may inform prevention strategies.

A Concurrent Extended Spectrum Beta-lactamase Production and Multidrug Resistance among Proteus Species isolated from Clinical samples of patients attending selected Hospitals in North Eastern Nigeria.

Proteus species are rod-shaped, Gram-negative bacteria that cause opportunistic infections in the urinary tract and occasionally in the gastrointestinal tract. They are implicated in infections like cystitis and pyelonephritis, particularly in immunocompromised individuals, and are frequently present in cases of asymptomatic bacteriuria. Herein, we aimed to investigate the co-occurrence of extended-spectrum beta-lactamase (ESBL) enzyme production and multidrug resistance (MDR) among Proteus spp. Isolated from patients attending selected hospitals in Northeastern Nigeria. A total of 1,500 clinical samples from consenting patients across six states in the Northeastern region of Nigeria were collected. The samples were cultured on Blood agar, and growth resembling that of Proteus species were again subcultured onto MacConkey agar to obtain discrete colonies, further confirmed using biochemical tests. Antibiotics susceptibility test was carried out for all isolates using the Kirby-Bauer disc diffusion method, coupled with a screening of the production of extended-spectrum beta-lactamase using the Combined Disc Diffusion Method. Of the 1500 samples collected, 144 yielded positive growth for Proteus spp., resulting in a prevalence rate of 9.60%. Among these Proteus isolates, three species were identified, with Proteus mirabilis (90.97%) being the most abundant, followed by Proteus vulgaris (8.33%) and Proteus penneri (0.70%). The Proteus isolates displayed significant resistance to β-lactam antibiotics, with a Mean ± SD of 96.64 ± 22.73. A substantial portion of the Proteus spp. Isolated exhibited multidrug resistance (87.89%), with Proteus mirabilis (82.27%) being the most prevalent MDR species. Moreover, about 71.0% of the Proteus spp were ESBL producers, with Proteus mirabilis (64.54%) being the most predominant. Furthermore, 67.38% of all isolates exhibited MDR and ESBL production, and Proteus mirabilis (62.41%) was the most significant among the three Proteus species. These findings highlight the occurrence of multidrug resistance and ESBL production among Proteus spp. in Northeastern Nigeria, with Proteus mirabilis particularly noteworthy. This information is crucial for guiding clinical decision-making, especially in managing infections caused by multidrug-resistant and ESBL-producing Proteus strains.

Antibiotic Susceptibility of Klebsiella pneumonia Isolates from Hospitalized Patients in Three Referral Medical Centers in Isfahan, Iran

: Background: Klebsiella pneumoniae (K. pneumoniae) is a major cause of nosocomial and community-acquired infections (CAIs). Understanding the antibiotic susceptibility of this bacterium is essential for selecting the most effective antibiotic for associated infections. Objectives: We investigated the sensitivity of K. pneumonia (KP) isolates obtained from inpatients in Isfahan, Iran. Methods: Organism identification and antibiotic susceptibility testing were conducted using standard tests. Stratification was performed based on extended spectrum beta-lactamase (ESBL) production and the source of infection acquisition (community versus hospital). Results: A total of 437 KP isolates were investigated, of which 5.5% were pan drug resistant (PDR), 56.8% were extensively drug resistant (XDR), and 18% were ESBL producers. The isolates were most susceptible to colistin (100%), aminoglycosides (62.8%), and carbapenems (55%). Nosocomial isolates showed lower sensitivity to most of the examined antibiotics. Conclusions: The high prevalence of PDR and XDR KP strains in hospitalized patients is a major problem in the studied area.

PCR Detection of Extended Spectrum β-Lactamase from Some Gram Negative Bacteria of Clinical Source

Beta-lactamase-producing microorganisms present distinct challenges in health care systems around the world as a result of the extensive utilization of broad-spectrum antibiotics. The objective of this research endeavor was to ascertain the antibiotic resistance profile and presence of β-lactamase genes in Gram-negative bacteria that had been previously identified and collected from laboratory work benches at the Department of Microbiology and Parasitology, UCH, located in Ibadan, South-West Nigeria. The Kirby-Bauer disc diffusion method was used to assess antibiotic susceptibility on all the isolates, and the double disc synergy test was used to validate extended spectrum beta-lactamase (ESBL) production phenotypically. The PCR techniques were used to detect β-lactamase genes. The sample distribution of the forty clinical isolates were collected from the work benches and the distribution was: urine (17), blood (9), wound (6), sputum (4), amniotic fluid (3) and tracheal aspirate (1). Klebsiella pneumoniae was the most prevalent organism with 17 isolates ( 42.5% ). Out of the 40 isolates that were obtained, 21 were ESBL producers and the distribution was as follows: Klebsiella pneumoniae (9), Escherichia coli (6), Enterobacter cloacae (2), Enterobacter aerogenes (1), Pseudomonas aeruginosa (1), Acinetobacter baumannii (1) and Hafnia alvei (1). The majority of the clinical isolates showed considerable resistance to antibiotic classes that were tested, with amoxicillin-clavulanate showing the highest resistance rate at 65%. Out of all the ESBL producers, 35.3% had blaTEM, 29.4% had blaCTX-M, 23.5% had blaSHV, and 11.8% had blaCMY. AmpC β-lactamases, Antibiotic resistance, Disc diffusion, Gram-negative bacteria, Polymerase chain reaction.

Extended spectrum beta-lactamase production and blaCTX-M gene in Escherichia coli and Klebsiella pneumoniae causing urinary tract infection at a tertiary care hospital in Nepal

Background and objective: Urinary tract infections (UTIs) are the most common bacterial infections where Escherichia coli (E. coli) and Klebsiella pneumoniae (K. pneumoniae) are the predominating pathogens. These pathogens have a high rate of antibiotic resistance and exhibit the production of extended-spectrum beta-lactamase (ESBL). This study investigated the antibiotic resistance pattern and ESBL production of E. coli and K. pneumoniae isolated from patients with UTIs attending a tertiary care hospital in Nepal by both phenotypic and genotypic techniques. Materials and methods: A cross-sectional study was performed where 4664 mid-stream urine specimens from suspected UTI cases were cultured. Isolated E. coli and K. pneumoniae were subjected to antibiotic susceptibility testing by Kirby Bauer disc diffusion method. Genotypic detection of blaCTX-M gene was performed using polymerase chain reaction (PCR). Results: Out of 4664 urine samples processed, 564 (12.1%) were positive for E. coli (475, 10.2%) and K. pneumonia (89, 1.9%). Out of the total 564 studied samples, 267 (47.3%) were MDR isolates (E. coli: 222, 46.7%; K. pneumoniae: 45, 50.6%) and 96 (17%) were positive for ESBL by double disc confirmatory test. Out of 24 ESBL positive E. coli and 6 K. pneumoniae, 19 (79.2%) and 3 (50%) respectively were positive for blaCTX-M gene. Conclusion: This study indicates high prevalence of MDR and ESBL producing E. coli and K. pneumoniae causing UTIs at an urban hospital setting in Nepal. July 2024; Vol. 18(2):002. DOI: https://doi.org/10.55010/imcjms.18.014 *Correspondence: Gaurab Pandey, Department of Medical Laboratory Science, Nobel College Affiliated to Pokhara University, Kathmandu, Nepal. Email: pandeygaurab67@gmail.com

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.

Antibiotic Resistance Profile of E. coli Isolates from Lettuce, Poultry Manure, Irrigation Water, and Soil in Kumasi, Ghana.

Inputs such as irrigation water and poultry manure used in lettuce cultivation have been found to be associated with antibiotic-resistant pathogens. The study assessed the antibiotic resistance profile of Escherichia coli isolated from lettuce, poultry manure, irrigation water, and soil in Kumasi. One hundred and fifty-six samples of lettuce, irrigation water, soil, and manure were collected from three farms over a seven-week cultivation period (seedlings to harvest stage). E. coli were enumerated using standard methods. 98% of the samples were positive for E. coli. Geometric means for lettuce, irrigation water, and soil ranged from 2.0 × 105 to 1.67 × 107 MPN/100 ml while that of manure ranged from 2.0 × 105 to 1.31 × 107 MPN/100 ml. Generally, the microbial load of all parameters on all farms across the weeks was significant and exceeded World Health Organization (WHO) and International Commission on Microbiological Specifications for Foods (ICMSF) standard recommendations for food. Using the Kirby Bauer method, antibiotic sensitivity testing was performed against 225 biochemically confirmed E. coli with twelve antibiotics. Relatively high resistance was recorded for some members of the beta-lactam class: meropenem: 94.2%, ampicillin: 91.9%, cefuroxime: 95.1%, ceftriaxone: 94.7%, and cefotaxime: 94.2%. Eighty of the isolates were screened for extended spectrum beta lactamase (ESBL) production using cefotaxime (CTX) and cefotaxime/clavulanic acid (CTX/CLA) discs and three showed positive: one each from poultry manure, irrigation water, and soil. Polymerase chain reaction (PCR) confirmed the presence of blaCTX-M gene. The occurrence of antibiotic-resistant E. coli in vegetables and their production environment is alarming and poses serious health threats to the general public. The presence of blaCTX-M gene in E. coli from a vegetable production site recorded for the first time in Ghana requires enforcement by regulatory bodies on the inappropriate use of antibiotics in the country.

PREVALENCE OF BLATEM, BLASHV, AND BLACTX-M GENES AMONG ESBL-PRODUCING ESCHERICHIA COLI ISOLATED FROM THE BLOOD SAMPLES OF ICUS PATIENTS OF UNIVERSITY HOSPITALS IN SANA'A CITY, YEMEN

Aim and Objective: With the emergence of organisms such as Enterobacteriaceae that produce extended-spectrum β-lactamase (ESBL), which are resistant to multiple medications (multidrug-resistance), concerns about how best to treat infections have significantly increased. The current study examined the molecular features of ESBL in clinical isolates of Escherichia coli that resulted in bloodstream infections as well as the pattern of antibiotic resistance to gather useful data on the infection's epidemiology among Yemeni ICU patients. Subjects and methods: A cross-sectional study was conducted on sepsis patients admitted in intensive care units at four hospitals in Sana'a, Yemen, between January, 2021 and April, 2022. Blood cultures were used on patients suspected of having sepsis. Standard laboratory procedures were then used to isolate and identify possible bacterial infections, and the disk diffusion method was used to test for microbial susceptibility. All strains were tested for Extended Spectrum Beta-Lactamase (ESBL) production using the Modified Double Disc Synergy Test (MDDST). Following analysis, polymerase chain reaction, β-lactamase genes (blaTEM, blaSHV, and blaCTX-M) were identified. Results: The results of the conventional PCR experiment revealed that 33.3% of the blaCTX-M genes, 0.0% of blaSHV, and 100% of blaTEM were present in the strains of ESBL-producing E. coli that were collected. It was discovered that the E. coli isolates' patterns of antibiotic resistance to 23 different antibiotics differed greatly. The bulk of the E. coli isolates were found to be multi-drug resistant (MDR). Furthermore, MDR characteristics were observed in 85% of E. coli isolates. Conclusion: Control and surveillance of antibiotic resistance depend on an understanding of the resistance genes and patterns of antimicrobial resistance of bacterial pathogens within a given geographic area. The current study's findings showed that MDR was very common. According to the current study's findings, TEM was significantly more common than other ESBL gene types. Peer Review History: Received: 7 November 2023; Revised: 12 December; Accepted: 6 January 2024, Available online: 15 January 2024 Academic Editor: Dr. A.A. Mgbahurike, University of Port Harcourt, Nigeria, amaka_mgbahurike@yahoo.com Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.5/10 Average Peer review marks at publication stage: 8.0/10 Reviewers: Dr. Rola Jadallah, Arab American University, Palestine, rola@aauj.edu Dr. Wadhah Hassan Ali Edrees, Hajja University, Yemen, edress2020@gmail.com

Extended Spectrum Beta Lactamase Profile of Citrobacter Species Isolated from Various Samples in the North East Coast of Tamil Nadu, India: A Cross-sectional Study

Introduction: Citrobacter species play a significant role in causing hospital-acquired infections, especially those affecting the urinary and respiratory systems, and they are commonly present in faeces. Healthcare systems are facing a major challenge due to the rise of antibiotic-resistant Gram-negative infections resistant to antibiotics. Aim: To identify the genotypic characterisation of the resistant gene blaTEM-1 in Citrobacter spp. obtained from a range of clinical samples. This investigation seeks to provide insights into the patterns of antibiotic resistance exhibited by these bacteria. Materials and Methods: This cross-sectional study was conducted between August 2021 and February 2022. Preliminary examinations included screening as well as confirmatory tests for Extended Spectrum Beta Lactamase (ESBL) production, which were confirmed by the combination disk method and double disc synergy test. Genotypic detection of the blaTEM-1 gene was done using the Polymerase Chain Reaction (PCR) technique. Demographic parameters like gender and age were assessed. Statistical analysis was done using the Statistical Package for the Social Sciences (SPSS) Software version 21.0. Results: Citrobacter koseri (66 isolates, 54.5%) was found to be the predominant species among the Citrobacter spp., followed by Citrobacter freundii (55 isolates, 45.5%). Urine samples were the primary source of Citrobacter spp. isolates, comprising 52 isolates (43%) of the total. In this study, the prevalence of ESBL producers was reported to be 49 isolates (77.8%) identified as ESBL producers. Tigecycline demonstrated a 100% effectiveness rate, followed by Piperacillin-Tazobactam (96% sensitivity) and Amikacin (95% sensitivity). Out of the 20 Citrobacter spp. isolates that tested positive for ESBL, 15 isolates (75%) were found to be positive for blaTEM-1. Conclusion: The results of this study provide insight into the changing patterns of antibiotic resistance in Citrobacter spp., with particular emphasis on ESBL profiles. Thus, it is imperative to implement intervention techniques to reduce the ongoing selection and spread of these more resistant bacteria, as well as efficient infection-control measures to manage epidemics. These results advance the comprehension of antibiotic resistance patterns and guide initiatives to combat the increasing threat of antibiotic resistance.

Investigation of Virulence Factors, Phylogenetic Group Features, and the Presence of ST131 Clone in Escherichia coli Isolates, a Urinary Tract Infection Agent in Children

Urinary tract infection (UTI) caused by Escherichia coli is a significant health issue in children. Today especially E.coli O25b/ST131, defined as a pandemic clone, is a serious public health problem due to its high virulence and antimicrobial resistance rates. In this study, a total of 200 (100 first and 100 recurrent UTI-causing) E.coli isolates from urine samples sent to the Ankara University School of Medicine Cebeci Training and Research Hospital Central Laboratory between January and September 2021 with the preliminary diagnosis of UTI in pediatric patients aged three to 18 years were analyzed for antimicrobial resistance rates, phylogenetic group distributions, virulence factor frequencies and whether they belong to the O25b/ST131 clone. It is aimed in this study that, the obtained data will shed light on new studies for diagnosis, treatment and prophylaxis options that can be developed for more effective UTI management by contributing to the surveillance studies in our country. Antimicrobial susceptibility of E.coli isolates identified by conventional methods was evaluated by Kirby-Bauer disc diffusion method and extended spectrum beta-lactamase (ESBL) production was evaluated by double disc synergy test. Polymerase chain reaction (PCR) was used for the investigation of phylogenetic grouping, the O25b/ST131 clone, virulence genes and the molecular level classification of the isolates detected as uropathogenic E.coli (UPEC). Pulsed-field gel electrophoresis (PFGE) was performed with the isolates collected at different times from the same patient. The highest antimicrobial resistance rates observed were against ampicillin (n= 100, 50%), cefazolin (n= 99, 49.5%), trimethoprim-sulfamethoxazole (n= 55, 27.5%), amoxicillin-clavulanic acid (n= 43, 21.5%) and cefotaxime (n= 43, 21.5%). In recurrent UTI agents, resistance rates were higher for cefotaxime (n= 29, 29%), trimethoprim-sulfamethoxazole (n= 35, 35%) and cefepime (n= 25, 25%) and in O25b/ST131 isolates (n= 67) the rates were higher for amikacin (n= 3, 4.5%), gentamicin (n= 10, 14.9%) and ciprofloxacin (n= 17, 25.4%) when compared to the first UTI agents and non-O25b/ ST131 isolates (p< 0.05). It was found that 29% (n = 58) of the isolates were multidrug resistant (MDR) and 19% (n = 38) produced ESBL.The rate of recurrent UTI agents was found to be higher among ESBL producing isolates and/or MDR isolates (n= 36, 62% and n= 27, 71%, respectively, p< 0.05). It was found that 45.5% (n= 91) of the isolates were in D, 37.5% (n= 75) in B2, 12.5% (n= 25) in A, and 4.5% (n= 9) in B1 phylogenetic groups and isolates belonging to B2 and D phylogenetic groups had higher antibiotic resistance rates and carried more virulence genes (p< 0.05). Of the isolates, 33.5% (n= 67) were found to belong to the O25b/ST131 clone, no significant difference was found between the O25b/ST131 rates among the first and recurrent UTI agents (p> 0.05). It was determined that the isolates most frequently carry virulence genes for adhesion [fimH 97% (n= 194), papA 57% (n= 114), yfcV 49.5% (n= 99)] and iron uptake systems [fyuA 85.5% (n= 171), chuA 78% (n= 156), iutA 73% (n= 146)]. All virulence factors were detected more frequently in isolates belonging to the O25b/ST131 clone (p< 0.05). Of the isolates, 97% (n= 65) belonging to the O25b/ST131 clone and 27.1% (n= 36) not belonging to this clone were defined as UPEC with molecular analysis (p< 0.0001). Thirty-three isolates belonging to 15 patients were evaluated with PFGE, and it was observed that the latter isolate and the first isolate of eight patients (53%) had the same band profile. Focusing on surveillance, diagnostic testing, treatment algorithms, and preventive measures for E.coli and especially for ST131 clone, which is frequently observed as causative agent in childhood UTIs, will help to manage challenging E.coli infections.

Phenotypic detection of extended spectrum beta-lactamase production in clinical and non - clinical isolates of klebsiella species of animal origins in Abeokuta, Nigeria.

Production of extended-spectrum β-lactamases (ESBLs) confer resistance to β-lactam antibiotics. This study investigated the presence of ESBL-producing Klebsiella spp in clinical and non-clinical samples from different animal species in Abeokuta, Nigeria. The species of Klebsiella were determined by biochemical characterization (Oxoid Microbact GNB 24E®), while phenotypic ESBL-production was confirmed by using the cefpodoxime and cefpodoxime/clavulanic acid combination disc kit. Ninety-five Klebsiella isolates were investigated in this study. Fifty-five (57.9%) of the isolates were obtained from the faeces of apparently healthy animals while 40 (42.1%) were from clinical samples. The Klebsiella isolates were identified as follows: K. oxytoca (34.7%), K. pneumoniae (26.3%), K. ozaenae (18.9%), K. terrigena (13.7%), K. rhinooscleromatis (5.3%), K. planticola (1.1%). Eight (8.4%) out of the 95 isolates of Klebsiella spp were identified as ESBL-producers. These included four isolates of Klebsiella pneumoniae, three of Klebsiella oxytoca and one of Klebsiella ozaenae. Five out of the eight ESBL-producing isolates were from clinical samples while three isolates were from the faeces of apparently healthy animals. The ESBL-producing isolates were 100% resistant to ampicillin, cefotaxime, ceftazidime, cefpodoxime, streptpmycin, tetracycline, compound sulfonamide, trimethoprim/sulphomethoxazole, and nalidixic acid. Apparently healthy carriers and sick animals can serve as sources of transmission of ESBL-producing Klebsiella spp to other animals and humans.

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 ......

Clinically significant and colonizing &lt;em&gt;Enterobacteria&lt;/em&gt; isolates with extended spectrum beta-lactamase production and their significance in horse breeding

This review article is devoted to the problem of the emergence and spread of antibiotic-resistant enterobacteria producing beta-lactamase in horse breeding. Foal colibacillosis is an acute infectious disease of young animals, manifested by diarrhea, signs of severe intoxication and dehydration of the body, caused by pathogenic Escherichia coli. Foals get sick in the first 3-7 days after birth. Infection occurs alimentary, less often – aerogenic and intrauterine transmission. The source of infection are sick and recovered animals. The duration of the disease is 2-3 days with a mortality rate of up to 90%, which leads to multiple economic losses. Multiresistant Escherichia coli can accumulate in the intestines of healthy animals and be excreted with fecal matter into the environment, which leads to the spread of pathogenic microflora among livestock. In this case, the animals are latent or hidden carriers that do not show clinical signs of the disease. Clinically significant isolates are causative agents of infectious and inflammatory diseases. These microorganisms represent a potential hazard for the development of an infectious process, they exhibit virulent properties that lead to damage to the tissues of the host organism, which significantly limits the possibility of their eradication. Along with clinically significant isolates, there are colonizing isolates that do not cause infection at a specific site. It should be noted that under certain conditions, the colonizing isolate can cause an infectious process. Irrational use of antimicrobials is a key reason for the development of tolerance. Also, the untimely detection of enzyme inactivation of antibiotics by laboratory methods and the spread of resistance genes play an important role. Currently, disks and media with antibacterial drugs are widely used to detect multidrug resistance of microorganisms. Along with these methods, PCR diagnostics and sequencing are used to accurately determine antibiotic resistance genes. Due to the importance and relevance of this topic, many scientists are conducting research to detect and study the multiresistance of enterobacteria such as E. coli, and their spread among animals.

P13 | Microbial evolution and growth arena (MEGA‐plate), accelerated evolution and co‐selection studies with Escherichia coli strains for extended spectrum beta‐lactamase (ESBL) production

Journal of Veterinary Pharmacology and TherapeuticsVolume 46, Issue S1 p. 91-91 POSTER PRESENTATIONS P13 | Microbial evolution and growth arena (MEGA-plate), accelerated evolution and co-selection studies with Escherichia coli strains for extended spectrum beta-lactamase (ESBL) production First published: 11 June 2023 https://doi.org/10.1111/jvp.13292Read the full textAboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL No abstract is available for this article. Volume46, IssueS1Special Issue: 15th International Congress of the European Association for Veterinary Pharmacology and Toxicology held Bruges, Belgium, July 2–5, 2023June 2023Pages 91-91 RelatedInformation

&lt;i&gt;In vitro&lt;/i&gt; activity of cefepime / sulbactam and biapenem against Enterobacterales and &lt;i&gt;Pseudomonas aeruginosa&lt;/i&gt; isolated from blood culture from patients with hematological diseases: results of a multicenter study

Background. Activity against Gram negative bacteria, including Pseudomonas aeruginosa, is required for first line antibiotic therapy in patients with febrile neutropenia.Aim. To study in vitro activity of cefepime / sulbactam and biapenem against Enterobacterales and P. aeruginosa strains in patients with bloodstream infection and hematologic diseases.Materials and methods. Susceptibility of cefepime / sulbactam and biapenem in comparison to anibiotics used for febrile neutropenia was studied among Escherichia coli (n = 100), Klebsiella pneumoniae (n = 100), Enterobacter cloacae complex (n = 30), and P. aeruginosa (n = 70) isolated from blood culture (2017–2021) from patients with hematological diseases and infection in 4 Russian hospitals. Activity was determined by broth microdilution method, interpretation was according to Clinical and Laboratory Standards Institute (CLSI, 2022) and European Committee on Antimicrobial Susceptibility Testing (EUCAST , 2022) criteria, for cefepime / sulbactam we used cefepime criteria. The values of the minimum inhibitory concentration (MIC), MIC50 and MIC90 were studied.Results. MIC90 of cefepime / sulbactam were lower in comparison with piperacillin / tazobactam for E. coli without extended spectrum beta-lactamase (ESBL) production (0.125 μg / mL vs 1 μg / mL), K. pneumoniae without ESBL-production (0.125 μg / mL vs 2 μg / mL), K. pneumoniae with ESBL-production (32 μg / mL vs 128 μg / mL) with comparable frequency of resistant strains. For P. aeruginosa, preference of cefepime / sulbactam over piperacillin / tazobactam were found both by lower MIC90 (8 μg / mL vs 32 μg / mL) and by lower frequency of resistant strains according to EUCAST criteria (4.3 % vs 25.7 %). The MIC90 values of cefepime / sulbactam compared to cefepime and ceftazidime were 4 times lower for K. pneumoniae with ESBL-production and for Enterobacter cloacea complex, 2–4 times lower for P. aeruginosa, 64 times lower for E. coli with ESBL production.Values of biapenem MIC90 for E. coli without and with ESBL-production (0.032 μg / mL) were in intermediate position between meropenem and imipenem; for K. pneumoniae without ESBL-production – identical to imipenem (0.064 μg / mL), for K. pneumoniae with ESBL – minimal (0.064 μg / mL) against imipenem and meropenem; for E. cloacae – comparable to meropenem (0.032 μg / mL). For P. aeruginosa without carbapenemase production, the MIC50 / MIC90 values of biapenem (0.125 / 16 μg / mL) were minimal compared to meropenem (0.25 / 64 μg / mL) and imipenem (0.5 / 64 μg / mL).Conclusion. The favorable in vitro activity of cefepime / sulbactam and biapenem are fully comply with the requirements for febrile neutropenia.

Monitoring of the Evolution of Antibiotic Resistance in Urinary Tract Infection at the Mohamed VI University Hospital in Marrakech

Introduction: Urinary tract infections are a public health problem due to the excess mortality and the additional costs they entail. Objective: To monitor the evolution of antibiotic resistance of uropathogenic germs over the last three years in patients treated at the CHU Med VI of MARRAKECH. Materials and Methods: Descriptive study carried out over a period of 3 years (from January 2018 to December 2020) including all urinary tract infections retained as meeting the criteria for microbiological and clinical infections in patients treated at the Mohammed VI University Hospital in Marrakech. Results: Out of a total of 37,394 ECBUs, the prevalence of UTI was 11% (n = 4275). The average age was 36 years, with a standard deviation of 25.9 years. The sex ratio M / F was 0.95. Uropathogenic germs were represented mainly by Enterobacteriaceae 78%. E. coli accounted for 66%. Enterobacteriaceae were resistant to third generation cephalosporins in 22% of cases, decreased sensitivity to Carbapenems in 7.2%, resistant to ciprofloxacin in 31%, to Cotrimoxazole in 42%, to Gentamicin in 17.5%, to Amikacin in 2%. Extended spectrum beta-lactamase production was 21%. Over the past three years, 895 multi-resistant bacteria have been identified, i.e. 23%. Klebsiella spp accounted for 41%. Conclusion: The results testify to the increased resistance of uropathogenic bacteria. Updated local studies are important to understand the state of play of bacterial ecology and resistance profiles in order to avoid treatment failures.

Plasmid-mediated quinolone resistance genes detected in Ciprofloxacin non-susceptible Escherichia coli and Klebsiella isolated from children under five years at hospital discharge, Kenya

BackgroundThe increasing spread of fluoroquinolone resistant enteric bacteria is a global public health concern. Children recently discharged from the hospital are at high risk of carriage of antimicrobial resistance (AMR) due to frequent exposure to antimicrobials during inpatient stays. This study aimed to determine the prevalence, correlates of ciprofloxacin (CIP) non-susceptibility, and distribution of plasmid-mediated quinolone resistance (PMQR) genes in Escherichia coli (E. coli) and Klebsiella spp isolated from children under five years being discharged from two Kenyan Hospitals.MethodsE. coli and Klebsiella spp were isolated from fecal samples from children discharged from hospital and subjected to antimicrobial susceptibility testing (AST) by disc diffusion and E-test. CIP non-susceptible isolates were screened for seven PMQR genes using multiplex polymerase chain reaction (PCR). Poisson regression was used to determine the association between the carriage of CIP non-susceptible isolates and patient characteristics.ResultsOf the 280 CIP non-susceptible isolates: 188 E. coli and 92 Klebsiella spp isolates identified among 266 discharged children, 195 (68%) were CIP-non-susceptible with minimum inhibitory concentrations (MICs) of ≥ 1 µg/mL. Among these 195 isolates, 130 (67%) had high-level CIP MIC = ≥ 32 µg/mL). Over 80% of the isolates had at least one PMQR gene identified: aac(6’)lb-cr (60%), qnrB (24%), oqxAB (22%), qnrS (16%), and qepA (6%), however, qnrA was not identified in any isolates tested. Co-carriage of qnrB with acc(6’)-lb-cr was the most predominant accounting for 20% of all the isolates. Ceftriaxone use during hospital admission and the presence of extended spectrum beta-lactamase (ESBL) production were significantly associated with the carriage of CIP non-susceptible E. coli and Klebsiella spp.ConclusionCIP non-susceptibility is common among E. coli and Klebsiella spp isolated from hospital discharged children in Kenya. Carriage and co-carriage of PMQR, including the newly identified qepA gene, were frequently observed. These findings suggest that children leaving the hospital may serve as an important reservoir for transmission of resistant E. coli and Klebsiella spp to the community. Enhanced surveillance for AMR determinants is critical to inform interventions to control antimicrobial-resistant bacteria.

Multidrug-resistant Escherichia coli and Klebsiella pneumoniae isolated from hospital sewage flowing through community sewage system and discharging into the Indian Ocean

BackgroundHospital sewage is a significant reservoir of antimicrobial-resistant pathogens and genes that pose a huge public health threat. In this study, we determined the occurrence of multidrug-resistant Escherichia coli and Klebsiella pneumoniae in sewage flowing from a referral hospital through the urban sewage system to the point of discharge in the Indian Ocean.ResultsA total of 400 sewage samples were collected, yielding 517 isolates. Of these, 32.3% (167/517) were from hospital sewage, while 67.7% (350/517) were from the community. E. coli was the most common isolate (44.5% (230/517)), followed by K. pneumoniae at 27.3% (141/517), and other gram-negative bacteria constituted 28.2% (146/517) of the isolates. Multidrug resistance (MDR) was seen in 80.9% (186/230) E. coli and 71.6% (101/141) K. pneumoniae. Of the MDR isolates, 27.2% (78/287) were resistant to four different classes of antibiotics, while 6.9% (20/287) exhibited resistance to eight classes. The most frequent MDR pattern was PEN/CEP/TET/QNL/SUL, seen in 14.2% (38/287) of the isolates. The isolation frequency of MDR E. coli and K. pneumoniae at different sampling sites was high, being 47.6% in hospital chambers, 62.0% in hospital ponds, 58.1% in the treated hospital wastewater, and 55.6% in the community stream draining into the Indian Ocean. Extended spectrum beta-lactamase production was observed in 40% (92/230) of E. coli and 36.2% (51/141) of K. pneumoniae isolates. Resistance to quinolones among E. coli was 54.8% (126/230) and was 39.7% in K. pneumoniae (56/141). Carbapenem resistance in E. coli was 39.6% (91/230), while among K. pneumoniae isolates was 32.6% (46/141).ConclusionsWe found high proportions of multidrug-resistant E. coli and K. pneumoniae in the wastewater flowing from the hospital through the community sewage system to the point where it enters the Indian Ocean. Biological treatment did not significantly reduce the proportion of resistant bacteria, posing a very serious public health threat. The release of these highly resistant pathogens into the Indian Ocean is of international concern.