Molecular Characterization of Escherichia coli Causing Urinary Tract Infections Through Next-Generation Sequencing: A Comprehensive Analysis of Serotypes, Sequence Types, and Antimicrobial and Virulence Genes. - Related Documents




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84801.0000Molecular Characterization of Escherichia coli Causing Urinary Tract Infections Through Next-Generation Sequencing: A Comprehensive Analysis of Serotypes, Sequence Types, and Antimicrobial and Virulence Genes. Introduction An enormous increase in antimicrobial resistance (AMR) among bacteria isolated from human clinical specimens contributed to treatment failures. Increased surveillance through next-generation sequencing (NGS) or whole genome sequencing (WGS) could facilitate the study of the epidemiology of drug-resistant bacterial strains, resistance genes, and other virulence determinants they are potentially carrying. Methods This study included 30 Escherichia coli (E. coli) isolates obtained from patients suffering from urinary tract infections (UTIs) attending Prathima Institute of Medical Sciences, Karimnagar, India. All bacterial isolates were identified, and antimicrobial susceptibility patterns were determined through conventional microbiological techniques and confirmed by automated systems. All the isolates were investigated using NGS to identify genes coding for resistance, such as extended-spectrum beta-lactamases (ESBLs), metallo-beta-lactamases, and virulence genes. Multilocus sequence typing (MLST) was used to understand the prevalent strain types, and serotyping was carried out to evaluate the type of O (cell wall antigen) and H (flagellar antigen) serotypes carried by the isolates. Results The conventional antimicrobial susceptibility testing revealed that 15 (50%) isolates were resistant to imipenem (IPM), 10 (33.33%) were resistant to amikacin (AK), 13 (43.33%) were resistant to piperacillin-tazobactam (PTZ), 17 (56.66%) were resistant to cephalosporins, and 14 (46.66%) were resistant to nitrofurantoin (NIT). Among the isolates, 26 (86.66%) had revealed the presence of multiple antibiotic-resistant genes with evidence of at least one gene coding for beta-lactamase resistance. There was a high prevalence of bla(CTX-M )(19/30, 63.33%) genes, followed by bla(TEM) and bla(OXA-1). The bla(NDM-5) gene was found in three isolates (3/30, 10%). The virulence genes identified in the present study were iutA, sat, iss, and papC, among others. The E. coli serotype found predominantly belonged to O25:H4 (5, 16.66%), followed by O102:H6 (4, 13.33%). A total of 16 MLST variants were identified among the examined samples. Of the MLST-based sequence types (STs) identified, ST-131 (7, 23.33%) was the predominant one, followed by ST-167 (3, 10%) and ST-12 (3, 10%). Conclusions The study results demonstrated that the E. coli strains isolated from patients suffering from UTIs potentially carried antimicrobial resistance and virulence genes and belonged to different strain types based on MLST. Careful evaluation of bacterial strains using molecular analyses such as NGS could facilitate an improved understanding of bacterial antibiotic resistance and its virulence potential. This could enable physicians to choose appropriate antimicrobial agents and contribute to better patient management, thereby preventing the emergence and spread of drug-resistant bacteria.202438576671
91710.9999Virulence characterization and clonal analysis of uropathogenic Escherichia coli metallo-beta-lactamase-producing isolates. BACKGROUND: Uropathogenic Escherichia coli (UPEC) is a major cause of urinary tract infection (UTI); however, treatment of UTI has been challenging due to increased antimicrobial resistance (AMR). One of the most important types of AMR is carbapenem resistance (CR). CR bacteria are known as an important threat to global public health today. Class B metallo-beta-lactamases (MBLs) are one of the major factors for resistance against carbapenems. We aimed to investigate the characteristics of UPEC isolates producing MBL. METHODS: A cross-sectional study was conducted from October 2018 to December 2019 in Ahvaz; Iran. UPEC isolates were identified by biochemical and molecular methods. Metallo-beta-lactamase-producing isolates were detected using modified carbapenem inactivation method (mCIM) and EDTA-CIM (eCIM) tests. MBL genes, phylogenetic group, and virulence genes profile of carbapenem resistant isolates were determined. Conjugation assay and plasmid profiling were conducted to evaluate the ability of transferring of CR to other E. coli isolates. Clonal similarity of isolates were assessed using Enterobacterial intergenic repetitive element sequence (ERIC)-PCR. RESULTS: Among 406 UPEC isolates, 12 (2.95%) carbapenem-resistant were detected of which 11 were phenotypically MBL-producing strains. Four isolates were resistant to all investigated antimicrobial agents and were considered possible pandrug-resistant (PDR). bla(NDM), bla(OXA-48), bla(IMP-1), and bla(IMP-2) genes were found in 9, 5, 1, and 1 isolates, respectively. Among 30 virulence genes investigated, the traT, fyuA followed by fimH, and iutA with the frequency of 8 (66.7%), 8 (66.7%), 7 (58.3%), and 7 (58.3%) were the most identified genes, respectively. Siderophore production was the main virulence trait among carbapenem-resistant UPEC isolates. Except for two, all other isolates showed weak to moderate virulence index. In all recovered isolates, CR was readily transmitted via plasmids to other isolates during conjugation experiments. CONCLUSION: MBL and carbapenemase genes, especially bla(NDM) and bla(OXA-48) are spreading rapidly among bacteria, which can be a threat to global public health. Therefore monitoring the emergence and dissemination of new AMR is necessary to continuously refine guidelines for empiric antimicrobial therapy. Understanding the mechanisms of resistance and virulence in this group of bacteria can play an effective role in providing new therapeutic methods.202134344363
98020.9998Phenotypic and Molecular Characterization of Extended-Spectrum β-Lactamase, Plasmid-Mediated- AmpC, and Carbapenemase-Producing Enterobacteriaceae Isolated from Companion and Production Animals in Brazil. The crisis of bacterial resistance is an emerging One Health challenge, driven by the overuse of antimicrobials in medical and agricultural settings. This study aimed to investigate extended-spectrum β-lactamase (ESBL), Ampicillinase (AmpC), and carbapenemase production, and the presence of genes encoding these enzymes in Escherichia coli, Klebsiella spp., and Proteus spp., major contributors to infections and resistance isolates from animals. From 2016 to 2021, 130 multidrug-resistant (MDR) or extensively drug-resistant (XDR) isolates were recovered from the secretions, excretions, and organs of companion and production animals with active infections. Antibacterial sensitivity tests, along with phenotypic and genotypic detection of resistance enzymes, were performed. To the best of our knowledge, this is the first study in Brazil to estimate the prevalence of XDR Enterobacteriales isolated from companion and production animals, which accounted for 13.8% of the strains. Statistically significant differences (P < 0.05) in resistant bacteria between different classes and within the same class of antibacterial bacteria were found. The statistical probability between genotypic detection of ESBL (OR = 3.1) and phenotypic tests for AmpC (OR = 2.3) was also established. Approximately 32.3%, 17.6%, and 16.8% of the strains had positive phenotypic tests for ESBL, AmpC, and carbapenemases, respectively. Genetic analysis revealed the presence of bla(CTX-M) (60.0%), bla(AmpC) (9.18%), bla(KPC-2) (0.76%), and bla(NDM) (1.52%). AmpC genes were identified in 8.46% of the samples, with bla(CMY) being the most frequent (6.92%), followed by bla(DHA) (0.77%), and bla(FOX) (0.77%). The sequenced amplicons were deposited in NCBI. This study reveals critical data on Enterobacteriaceae with antibacterial resistance genes isolated from animals and may pose a significant threat to One health.202539903315
116330.9998A Three-Year Look at the Phylogenetic Profile, Antimicrobial Resistance, and Associated Virulence Genes of Uropathogenic Escherichia coli. Uropathogenic Escherichia coli is the most common cause of urinary tract infections, resulting in about 150 million reported annual cases. With multidrug resistance on the rise and the need for global and region surveillance, this investigation looks at the UPEC isolates collected for a 3-year period, with a view of ascertaining their antimicrobial susceptibility patterns and associated virulence determinants. The identification of bacteria isolates, antimicrobial susceptibility, and extended-spectrum beta-lactamases (ESBLs) production was determined with a Vitek 2 Compact Automated System (BioMerieux, Marcy L'Etoile, France). ESBLs were confirmed by the combined disc test (CDT) and basic biochemical test. The isolates were distributed into A (11%), B1 (6%), B2 (62.4%), and D (20.6%). Resistance to the penicillin group was high, between 88% and 100%. Additionally, resistance was high to cephalosporins (100%) in 2017 and 2018. The isolates were all sensitive to tigecycline, while resistance against imipenem and meropenem was low, at 4-12% in 2017 and 2018 and 0% in 2019. The results also showed that ESBL isolates were seen in 2017 and 2018. They were confirmed positive to CTX/CLA (88.5%) and CAZ/CLA (85%). By 2019, the number of resistant isolates reduced, showing only 4% ESBL isolates. Two virulence genes, fimH (46%) and papE/F (15%), were detected among the isolates by PCR. In conclusion, this study found that phylogroups B2 and D carried the most virulence genes as well as MDR and ESBL characteristics, suggesting the UPEC strains to be extraintestinal pathogens responsible for UTIs.202235745485
297440.9998Diversity of Virulence Genes in Multidrug Resistant Escherichia coli from a Hospital in Western China. BACKGROUND: Escherichia coli strains are the most commonly isolated bacteria in hospitals. The normally harmless commensal E. coli can become a highly adapted pathogen, capable of causing various diseases both in healthy and immunocompromised individuals, by acquiring a combination of mobile genetic elements. Our aim was to characterize E. coli strains from a hospital in western China to determine their virulence and antimicrobial resistance potential. METHODS: A total of 97 E. coli clinical isolates were collected from the First Affiliated Hospital of Chengdu Medical College from 2015 to 2016. Microbiological methods, PCR, and antimicrobial susceptibility tests were used in this study. RESULTS: The frequency of occurrence of the virulence genes fimC, irp2, fimH, fyuA, lpfA, hlyA, sat, and cnf1 in the E. coli isolates was 93.81, 92.78, 91.75, 84.54, 41.24, 32.99, 28.86, and 7.22%, respectively. Ninety-five (97.9%) isolates carried two or more different virulence genes. Of these, 44 (45.4%) isolates simultaneously harbored five virulence genes, 24 (24.7%) isolates harbored four virulence genes, and 17 (17.5%) isolates harbored six virulence genes. In addition, all E. coli isolates were multidrug resistant and had a high degree of antimicrobial resistance. CONCLUSION: These results indicate a high frequency of occurrence and heterogeneity of virulence gene profiles among clinical multidrug resistant E. coli isolates. Therefore, appropriate surveillance and control measures are essential to prevent the further spread of these isolates in hospitals.201931824179
101650.9998Investigation of CTX-M Type Extended-Spectrum β-Lactamase, Carbapenem and Colistin Resistance in Enterobacterales Isolated From Dairy Cattle in Turkey. BACKGROUND: The increasing prevalence of antimicrobial resistance in animals, particularly the spread of multidrug-resistant Enterobacterales, poses a significant zoonotic and public health risk. OBJECTIVE: The aim of this study was to investigate extended-spectrum β-lactamase (ESBL), carbapenem and colistin resistance among Enterobacterales in faecal swabs of dairy cattle. METHODS: A total of 400 samples were cultured on Mac Conkey screening media for ESBL, carbapenem and colistin resistance. The grown Enterobacterales were identified by MALDI-TOF-MS, followed by ceftriaxone, cefotaxime and ceftazidime resistance and double disk synergy. ESBL resistance genes were identified by polymerase chain reaction (PCR) and Sanger sequencing. Bacteria grown on colistin screening media were investigated for colistin resistance by EUCAST microbroth dilution method. RESULTS: A total of 89 (22.25%) of the bacteria grown from 400 samples were identified as potential ESBL-producing Enterobacterales members. A number of 53 (59.5%) of them were identified as ESBL blaCTX-M as a result of PCR, and 10 of them were identified as blaCTX-M-15/28/36/66 as a result of sequencing. None of the samples cultured on carbapenem medium grew. A total of 18 samples grown in colistin medium were found to be colistin sensitive by broth microdilution. Genotypes were not included in the study. All isolated bacteria were identified as Escherichia coli. SOLUTION: In this study, blaCTX-M-15 and its derivatives, which are common in humans, were also found to be the predominant ESBL type in animals. Monitoring resistance in animals together with resistance in human infections may provide more important data on the spread of resistance.202540704983
101760.9998Evaluation of canine raw food products for the presence of extended-spectrum beta-lactamase- and carbapenemase-producing bacteria of the order Enterobacterales. OBJECTIVE: To assess the potential contamination of commercial raw dog food products with bacteria of the Enterobacterales order that produce extended spectrum beta-lactamase (ESBL) and carbapenemase enzymes, determine risk factors for contamination, and understand isolate genetic diversity. SAMPLES: A total of 200 canine raw food products. METHODS: Products were cultured on selective chromogenic agar following enrichment steps. Whole-genome sequencing was performed for isolates that were confirmed to produce an ESBL. Isolates were characterized by antimicrobial resistance genes, and multilocus sequences typing, and compared to other isolates in the NCBI database for clonality. Preservation method and protein sources were assessed as potential risk factors for contamination with ESBL and carbapenemase-producing bacteria of the Enterobacterales order. RESULTS: No carbapenemase-producing Enterobacterales (CPE) were identified, but ESBL-producing Enterobacterales bacteria were isolated from 20/200 products (10.0%; 95% CI, 7.3 to 16.5%), all of which were frozen. Pork-derived protein source products were 8.1 times (P = .001; 95% CI, 2.53 to 26.2) more likely to carry ESBL-producing Enterobacterales bacteria than other protein sources. WGS analysis confirmed the presence of ESBL genes in a total of 25 distinct isolates (19 Escherichia coli, 5 Klebsiella pneumoniae, and 1 Citrobacter braakii). Genes encoding CTX-M type ESBL enzymes were the most common (24/25 isolates, 96.0%) with blaCTX-M-27 being the most common allele (8/25, 32.0%). CLINICAL RELEVANCE: Frozen, raw food products may serve as a route of transmission of ESBL-producing Enterobacterales bacteria to companion animals. Veterinarians should advise owners about the risks of raw food diets, including potential exposure to antimicrobial-resistant bacteria.202235895774
91970.9998Molecular Characteristics of Carbapenem-Resistant Enterobacter cloacae in Ningxia Province, China. The emergence of carbapenem-resistant Enterobacteriaceae (CRE) has become a major public health concern worldwide and a new challenge in the treatment of infectious diseases. The molecular characteristics of Enterobacter cloacae in Ningxia China are unknown. In this study, we reported 10 carbapenem-resistant E. cloacae isolates from the General Hospital of Ningxia Medical University, the largest university hospital in Ningxia between January 2012 and December 2013. Bacteria isolates were identified by Vitek2 compact and the identity of non-duplicate E. cloacae isolates was further confirmed by PCR and sequencing. The drug susceptibility and phenotype identification of these isolates were analyzed by agar dilution method, modified Hodge test (MHT), and EDTA synergy test. Beta-lactamase (bla) genes bla(NDM-1) was found in 8 out of 10 isolates. Most isolates harbored multiple resistance genes including bla(ESBL), bla(AmpC), quinolones, aminoglycosides, and disinfectant resistance genes. Pulsed field gel electrophoresis (PFGE) showed that these E. cloacae isolates were grouped into 6 clusters based on a cutoff of 80% genetic similarity. In conjugative assay, 9 out of 10 isolates transferred carbapenem-resistant genes to Escherichia coli. Our study has revealed that NDM-1-producing isolates are the most prevalent carbapenem-resistant E. cloacae in Ningxia. These isolates also carry several other carbapenem-resistant genes and can transfer these genes to other bacteria through conjugation. These findings highlight an urgent need to monitor these isolates to prevent their further spread in this region.201728197140
114580.9998Abundance of Mobilized Colistin Resistance Gene (mcr-1) in Commensal Escherichia coli from Diverse Sources. Aims: Antimicrobial resistance (AMR) spreads not only by pathogenic but also by commensal bacteria, and the latter can become a reservoir for resistance genes. This study was aimed to investigate the AMR patterns along with the presence of mobilized colistin resistance (mcr) genes in commensal Escherichia coli circulating in chickens, farm environments, street foods, and human patients. Materials and Methods: By a cross-sectional survey, isolates obtained from 530 samples were tested for their AMR profiles against 9 antimicrobials. Minimum inhibitory concentration (MIC) of the phenotypically colistin-resistant isolates was determined and screened for a set of mcr genes followed by sequencing of mcr-1 gene in the multidrug-resistant (MDR) isolates. Results: A total of 313 E. coli strains were isolated and confirmed by polymerase chain reaction. Antimicrobial susceptibility testing revealed that about 98% (confidence interval [95% CI] 95-99) of the isolates were MDR, and 58% (95% CI 52-63) isolates exhibited resistance to colistin. MIC values of colistin against the isolates ranged from 4 to 64 mg/L. Except for human patients, 20.4% colistin-resistant isolates from other sources of isolation had mcr-1 gene. Conclusions: There is abundance of commensal MDR E. coli strains with the acquisition of mcr-1 gene circulating in chickens and farm environments in Bangladesh.202133909471
92490.9998Screening of Antimicrobial Resistance Genes and Epidemiological Features in Hospital and Community-Associated Carbapenem-Resistant Pseudomonas aeruginosa Infections. INTRODUCTION: Researching carbapenem-resistant isolates enables the identification of carbapenemase-producing bacteria and prevents their spread. METHODS: P. aeruginosa isolates were recovered from Medicine Faculty of Recep Tayyip Erdoğan University and identified by conventional methods and the automated Vitek 2 Compact system. Antimicrobial susceptibility experiments were performed in accordance with CLSI criteria and the automated Vitek 2 Compact system. The PCR method was investigated for the presence of β-lactamase resistance genes. PFGE typing was performed to show clonal relation among samples. RESULTS: Seventy P. aeruginosa isolates were isolated from seventy patients. Of the patients, 67.1% had contact with the health service in the last 90 days and 75.7% of the patients had received antimicrobial therapy in the previous 90 days. Twenty-four isolates were carbapenem resistant, 2 isolates were multidrug-resistant except colistin, and none of the samples had colistin resistance. The gene encoding β-lactamase or metallo-β-lactamase was found in a total of 36 isolates. The bla (VEB) and bla (PER) genes were identified in 1 and 5 isolates alone or 17 and 13 isolates in combination with other resistance genes, respectively. The bla (NDM) was the most detected metallo-β-lactamase encoding gene (n=18), followed by bla (KPC) (n=12). bla (IMP) and bla (VIM) were detected in 5 and 1 isolates, respectively. Also, the association of bla (VEB)-bla (PER) and bla (VEB)-bla (KPC)-bla (NDM) was found to be very high. Much more resistance genes and co-occurrence were detected in hospital-acquired samples than community-acquired samples. No difference was found between the community and hospital-associated isolates according to PFGE results. Simultaneously from 6 patients, other microorganisms were also isolated and 5 of them died. CONCLUSION: The average length of stay (days) was found to be significantly higher in HAI group than CAI group. The death of 5 patients with fewer or no resistance genes showed that the co-existence of other microorganisms in addition to resistance genes was important on death.202133907430
1051100.9998Multi-drug Resistance, β-Lactamases Production, and Coexistence of bla (NDM-1) and mcr-1 in Escherichia coli Clinical Isolates From a Referral Hospital in Kathmandu, Nepal. The ability of pathogenic Escherichia coli to produce carbapenemase enzymes is a characteristic that allows them to resist various antibiotics, including last-resort antibiotics like colistin and carbapenem. Our objectives were to identify rapidly developing antibiotic resistance (AR), assess β-lactamases production, and detect mcr-1 and bla (NDM-1) genes in the isolates. A prospective cross-sectional study was carried out in a referral hospital located in Kathmandu from November 2019 to December 2020 using standard laboratory and molecular protocols. Among 77 total E. coli isolates, 64 (83.1%) of them were categorized as MDR. Phenotypically 13 (20.3%) colistin-resistant, 30 (46.9%) ESBL and 8 (12.5%) AmpC producers, and 5 (7.8%) ESBL/AmpC co-producers were distributed among MDR-E. coli. Minimum inhibitory concentrations (MIC) against the majority of MDR isolates were exhibited at 1 g/L. Of these 77 E. coli isolates, 24 (31.2%) were carbapenem-resistant. Among these carbapenem-resistant bacteria, 11 (45.9%) isolates were reported to be colistin-resistant, while 15 (62.5%) and 2 (8.3%) were MBL and KPC producers, respectively. Out of 15 MBL producers, 6 (40%) harbored bla (NDM-1), and 8 (61.5%) out of 13 colistin-resistant pathogens possessed mcr-1. The resistance by colistin- and carbapenem were statistically associated (P < .001). However, only 2 (18.2%) of the co-resistant bacteria were found to have both genes. Our study revealed the highly prevalent MDR and the carbapenem-resistant E. coli and emphasized that the pathogens possess a wide range of capabilities to synthesize β-lactamases. These findings could assist to expand the understanding of AR in terms of enzyme production.202336741474
866110.9998Opening Pandora's box: High-level resistance to antibiotics of last resort in Gram-negative bacteria from Nigeria. OBJECTIVES: The aim of this study was to determine the percentage of antimicrobial-resistant isolates and the associated resistance mechanisms in Gram-negative bacteria from South Western Nigeria. METHODS: A total of 306 non-duplicate unbiased Gram-negative isolates were recovered from patients admitted to three teaching hospitals in South Western Nigeria in 2011 and 2013. Isolates were from clinical samples as well as from stool samples of inpatients without infection to assess antimicrobial resistance patterns in carriage isolates. Antimicrobial susceptibility testing was performed, and PCR and sequencing were used to identify genes encoding various known β-lactamases. Based on phenotypic and genotypic results, 10 isolates representing the diversity of phenotypes present were selected for whole-genome sequencing (WGS). RESULTS: Antimicrobial susceptibility testing revealed the following resistance rates: fluoroquinolones, 78.1%; third-generation cephalosporins, 92.2%; and carbapenems, 52.6%. More resistant isolates were isolated from stools of uninfected patients compared with clinical infection specimens. Klebsiella (10%) and Escherichia coli (7%) isolates produced a carbapenemase. WGS of selected isolates identified the presence of globally disseminated clones. CONCLUSION: This study illustrates a crisis for the use of first-line antimicrobial therapy in Nigerian patients. It is likely that Nigeria is playing a significant role in the spread of antimicrobial resistance owing to its large population with considerable global mobility.202031654790
867120.9998Epidemiology and Mechanism of Drug Resistance of Multidrug-Resistant Klebsiella Pneumoniae Isolated from Patients with Urinary Tract Infection in Beijing Teaching Hospital, China. PURPOSE: Klebsiella pneumoniae is an important pathogenic bacterium in causing urinary tract infection. With the overuse of antibiotics, bacteria resistant to quinolones combined with carbapenems are increasing. In this study, we investigated the epidemiology, molecular characteristics, drug resistance of multidrug-resistant Klebsiella pneumoniae (MDR-KPN) isolated from urine samples. It provides theoretical basis for the treatment of urinary tract infection by clinicians. PATIENTS AND METHODS: Fifty-one strains of Klebsiella pneumonia were obtained from urine samples collected between 2012 and 2017 in total. All the strains are multi-drug resistant bacteria. This paper used multilocus sequence typing (MLST) to determine molecular epidemiological typing. We performed antimicrobial susceptibility testing and investigated quinolones and carbapenems resistance genes. RESULTS: The strains which we collected were resistant to ciprofloxacin and Levofloxacin. In an epidemiological analysis using MLST, 86.27% (44/51) of isolates were confirmed to be ST11. The main carbapenem resistance gene was KPC-19, 78.43(40/51). Among the quinolone resistance genes, the major resistance genes were aac(6')-Ib-cr, oqxA and oqxB. CONCLUSION: The main molecular epidemiological types we detected was ST11. The main resistance gene of carbapenems was KPC-19. The quinolone resistance genes are mainly aac(6')-Ib-cr, oqxA and oqxB. The experimental results can help control the use of quinolones and carbapenems, and we could provide rational drug use basis for clinicians to treat urinary tract infection. For MDR-KPN, a combination of multiple antibiotics is necessary.202539803309
2151130.9998Study of the Genomic Characterization of Antibiotic-Resistant Escherichia Coli Isolated From Iraqi Patients with Urinary Tract Infections. Urinary tract infection is one of the last diseases prevalent in humans, with various causative agents affecting 250 million people annually, This study analyzed UTIs in Iraqi patients caused by Escherichia coli. ESBL enzymes contribute to antibiotic resistance. The research aimed to analyze ESBL gene frequency, resistance patterns, and genetic diversity of E. coli strains; Between Dec 2020 and May 2021, 200 urine samples were collected, cultured on blood agar, EMB, and MacConkey's plates, samples incubated at 37 °C for 24 h. Positive samples (> 100 cfu/ml) underwent Kirby-Bauer and CLSI antibiotic susceptibility testing. PCR detected virulence genes, Beta-lactamase coding genes, and biofilm-associated resistance genes in E. coli isolates; Out of 200 isolates, 80% comprised Gram-positive and Gram-negative bacteria. Specifically, 120 isolates (60%) were Gram-negative, while 40 isolates (20%) were Gram-positive. Among Gram-negative isolates, 20% were identified as E. coli. Remarkably, all E. coli strains showed resistance to all tested antibiotics, ranging from 80 to 95% resistance. The E. coli isolates harbored three identified resistance genes: blaTEM, blaSHV, and blaCTXM. Regarding biofilm production, 10% showed no formation, 12% weak formation, 62% moderate formation, and 16% strong formation; our study found that pathogenic E. coli caused 20% of UTIs. The majority of studied E. coli strains from UTI patients carried the identified virulence genes, which are vital for infection development and persistence.202439011020
2150140.9998Analysis of drug resistance genes of integrons in clinical isolates of Escherichia coli from elderly bloodstream infections. This experiment was carried out to provide a basis for the treatment of clinical bloodstream infections by analyzing the drug resistance characteristics and integrated gene distribution of Escherichia coli in bloodstream infections in elderly patients. For this aim, E. coli were collected for bacterial identification and drug sensitivity testing from bloodstream infections in elderly patients in the hospital from January 2016 to December 2019. ESBLs positive strains were assayed for genotypes and their integron carriage rates by PCR amplification. The characteristics and differences of various genotype rates were compared and analyzed. Results showed that a total of 230 E. coli strains were isolated. The detection rate of ESBLs-producing bacteria was 37.39 %. ESBLs-producing E. coli showed a high rate of resistance to cefepime, levofloxacin, cotrimoxazole, and ticarcillin/clavulanic acid (>40%). The resistance rate of 230 strains of E. coli to meropenem, minocycline, amikacin, gentamicin and cefoxitin was less than 20%. Among the ESBLs-producing E. coli in bloodstream infections in elderly patients, CTX-M-9 accounted for 27.91%, CTX-M-2 for 17.44%, and SHV for 13.95%. The detection rate of type I integrated genes was 41.30%, and type II and III integrated genes were not detected. ESBLs-producing genotyping-positive bacteria were detected with more than 50% of type I integrated genes. It was concluded that type I integrated genes in ESBLs-producing E. coli isolated from elderly patients carried resistance genes such as CTX-M-9 and CTX-M-2 aggravating multi-drug resistance in bacteria.202236227675
979150.9998Integrative phenotypic and genomic analysis of extended-spectrum Beta-lactamase (ESBL) and carbapenemase genes in Enterobacteriaceae and Pseudomonaceae strains isolated from animals in a Spanish Veterinary Teaching Hospital. Antimicrobial resistance (AMR) is a major global health threat, exacerbated by globalization which facilitates the spread of resistant bacteria. Addressing this issue requires a One Health perspective, involving humans, animals, and the environment. This study aims to compare the phenotypic resistance profiles of 69 clinical bacterial isolates (Enterobacteriaceae and Pseudomonaceae) from a Veterinary Teaching Hospital in Spain with their genotypic resistance profiles based on the presence of Extended-Spectrum Beta-Lactamases (ESBLs), AmpC and carbapenemases -enconding genes. For the genotypical analysis, whole genome sequencing (WGS) was used. Phenotypic characterization revealed that 37 isolates (53.6 %) grew on ESBL-selective medium. Phenotypic confirmatory tests showed that 12 strains (17.4 %) had some type of ESBL and 21 (30.4 %) could have an AmpC. Also, 24 isolates (34.8 %) grew in selective media for carbapenemases-producing bacteria, and 2 of these had a class A carbapenemase based on the KPC&MBL&OXA-48 disc kit. The genotypic analysis revealed 20 isolates (29 %) had bla(TEM), 8 (11.6 %) had bla(CTX-M) and 7 (10.1 %) bla(SHV). 27 (39.1 %) isolates had class C beta-lactamase genes. 35 isolates (50.7 %) had bla(OXA), class D beta-lactamase. 37 strains (53.6 %) had an Inc. plasmid replicon associated with the spread of AMR genes, including beta-lactamases and carbapenemases. This study emphasizes the value of combining phenotypic and genomic analyses to better understand and address antibiotic resistance, especially in veterinary contexts. Integrating these approaches enhances diagnostic accuracy by identifying strains with resistance genes that may not show phenotypically, helping clinicians in anticipating resistance under selective pressure.202539808975
883160.9998Detection of plasmid-mediated colistin resistance, mcr-1 gene, in Escherichia coli isolated from high-risk patients with acute leukemia in Spain. BACKGROUND: Bacterial infections in immunocompromised patients are associated with a high mortality and morbidity rate. In this high-risk group, the presence of multidrug-resistant (MDR) bacteria, particularly bacteria that harbor a transferable antibiotic resistance gene, complicates the management of bacterial infections. In this study, we investigated the presence of the transferable colistin resistance mcr genes in patients with leukemia in Spain. METHODS: 217 fecal samples collected in 2013-2015 from 56 patients with acute leukemia and colonized with MDR Enterobacteriaceae strains, were screened on September 2017 for the presence of the colistin resistance mcr genes (mcr-1 to -5) by multiplex PCR. mcr positive strains selected on LBJMR and MacConkey supplemented with colistin (2 μg/ml) media were phenotypically and molecularly characterized by antimicrobial susceptibility testing, minimum inhibitory concentration, multilocus sequence typing and plasmid characterization. RESULTS: Among 217 fecal samples, 5 samples collected from 3 patients were positive for the presence of the mcr-1 colistin-resistance gene. Four Escherichia coli strains were isolated and exhibited resistance to colistin with MIC = 4 μg/ml. Other genes conferring the resistance to β-lactam antibiotics have also been identified in mcr-1 positive strains, including bla(TEM-206) and bla(TEM-98). Three different sequence types were identified, including ST1196, ST140 and ST10. Plasmid characterization allowed us to detect the mcr-1 colistin resistance gene on conjugative IncP plasmid type. CONCLUSION: To the best of our knowledge, we have identified the mcr-1 gene for the first time in leukemia patients in Spain. In light of these results, strict measures have been implemented to prevent its dissemination.201931023570
1647170.9998Genomic and antimicrobial resistance genes diversity in multidrug-resistant CTX-M-positive isolates of Escherichia coli at a health care facility in Jeddah. BACKGROUND: Whole genome sequencing has revolutionized epidemiological investigations of multidrug-resistant pathogenic bacteria worldwide. Aim of this study was to perform comprehensive characterization of ESBL-positive isolates of Escherichia coli obtained from clinical samples at the King Abdulaziz University Hospital utilizing whole genome sequencing. METHODS: Isolates were identified by MALDI-TOF mass spectrometry. Genome sequencing was performed using a paired-end strategy on the MiSeq platform. RESULTS: Nineteen isolates were clustered into different clades in a phylogenetic tree based on single nucleotide polymorphisms in core genomes. Seventeen sequence types were identified in the extended-spectrum β-lactamase (ESBL)-positive isolates, and 11 subtypes were identified based on distinct types of fimH alleles. Forty-one acquired resistance genes were found in the 19 genomes. The bla(CTX-M-15) gene, which encodes ESBL, was found in 15 isolates and was the most predominant resistance gene. Other antimicrobial resistance genes (ARGs) found in the isolates were associated with resistance to tetracycline (tetA), aminoglycoside [aph(3″)-Ib, and aph(6)-Id], and sulfonamide (sul1, and sul2). Nonsynonymous chromosomal mutations in the housekeeping genes parC and gyrA were commonly found in several genomes. CONCLUSION: Several other ARGs were found in CTX-M-positive E. coli isolates confer resistance to clinically important antibiotics used to treat infections caused by Gram-negative bacteria.202031279801
1142180.9998Virulence Determinants and Plasmid-Mediated Colistin Resistance mcr Genes in Gram-Negative Bacteria Isolated From Bovine Milk. A major increase of bacterial resistance to colistin, a last-resort treatment for severe infections, was observed globally. Using colistin in livestock rearing is believed to be the ground of mobilized colistin resistance (mcr) gene circulation and is of crucial concern to public health. This study aimed to determine the frequency and virulence characteristics of colistin-resistant Gram-negative bacteria from the milk of mastitic cows and raw unpasteurized milk in Egypt. One hundred and seventeen strains belonging to Enterobacteriaceae (n = 90), Pseudomonas aeruginosa (n = 10), and Aeromonas hydrophila (n = 17) were screened for colistin resistance by antimicrobial susceptibility testing. The genetic characteristics of colistin-resistant strains were investigated for mcr-1-9 genes, phylogenetic groups, and virulence genes. Moreover, we evaluated four commonly used biocides in dairy farms for teat disinfection toward colistin-resistant strains. Multidrug-resistant (MDR) and extensive drug-resistant (XDR) phenotypes were detected in 82.91% (97/117) and 3.42% (4/117) of the isolates, respectively. Of the 117 tested isolates, 61 (52.14%) were colistin resistant (MIC >2 mg/L), distributed as 24/70 (34.29%) from clinical mastitis, 10/11 (90.91%) from subclinical mastitis, and 27/36 (75%) from raw milk. Of these 61 colistin-resistant isolates, 47 (19 from clinical mastitis, 8 from subclinical mastitis, and 20 from raw milk) harbored plasmid-borne mcr genes. The mcr-1 gene was identified in 31.91%, mcr-2 in 29.79%, mcr-3 in 34.04%, and each of mcr-4 and mcr-7 in 2.13% of the colistin-resistant isolates. Among these isolates, 42.55% (20/47) were E. coli, 21.28% (10/47) A. hydrophila, 19.12% (9/47) K. pneumoniae, and 17.02% (8/47) P. aeruginosa. This is the first report of mcr-3 and mcr-7 in P. aeruginosa. Conjugation experiments using the broth-mating technique showed successful transfer of colistin resistance to E. coli J53-recipient strain. Different combinations of virulence genes were observed among colistin-resistant isolates with almost all isolates harboring genes. Hydrogen peroxide has the best efficiency against all bacterial isolates even at a low concentration (10%). In conclusion, the dissemination of mobile colistin resistance mcr gene and its variants between MDR- and XDR-virulent Gram-negative isolates from dairy cattle confirms the spread of mcr genes at all levels; animals, humans, and environmental, and heralds the penetration of the last-resort antimicrobial against MDR bacteria. Consequently, a decision to ban colistin in food animals is urgently required to fight XDR and MDR bacteria.202134888259
1059190.9998Dissemination and phenotypic characterization of ESBL-producing Escherichia coli in Indonesia. BACKGROUND: The alarming rise in infections caused by extended-spectrum beta-lactamase (ESBL)-producing Escherichia coli in animals and humans poses a serious threat due to its escalating antibiotic resistance. Unveiling this problematic bacteria's prevalence and resistance patterns in animals is crucial for formulating effective control strategies and safeguarding public health. AIM: The purpose of this study was to analyze the expression of three main genes: blaCTX-M, blaSHV, and blaTEM, in ESBL-producing E. coli isolates from The Research Center for Veterinary Science and the National Research and Innovation Agency. Moreover, their resistance profiles against various antibiotics should be systematically evaluated. METHODS: Ninety-seven E. coli isolates from the bacteriology laboratory of The Research Center for Veterinary Science were identified on MacConkey medium supplemented with cefotaxime. The isolates were verified for the existence of the blaCTX-M, blaSHV, and blaTEM genes using PCR. Antimicrobial susceptibility testing was conducted using antibiotic discs following the CLSI standards. RESULTS: The prevalence of ESBL-producing E. coli in chicken ceca, eggs, and fish intestines was 16.5% (16/97). The specific genes detected were blaCTX-M gene at 93.75% (15/16), followed by the blaTEM gene, at 81.25% (13/16), and blaSHV at 25% (4/16). The antimicrobial sensitivity test results revealed that all ESBL-producing E. coli isolates had multidrug resistance 81.25% to 1-5 antibiotics and 18.75% to 6-7 antibiotics. The isolate exhibited 100% resistance to ampicillin and sulfamethoxazole, with exclusive sensitivity to chloramphenicol. CONCLUSION: The dominant gene in the ESBL-producing isolates was blaCTX-M. This bacterium is completely resistant to ampicillin and sulfamethoxazole, whereas it displays multidrug resistance to 1-7 different types of antibiotics.202540276175