Uropathogenic Escherichia Coli Isolates Research Articles

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

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

Whole genome sequencing of uropathogenic E. coli from Ireland reveals diverse resistance mechanisms and strong correlation with phenotypic (EUCAST) susceptibility testing

Urinary tract infections (UTI) caused by uropathogenic Escherichia coli (UPEC) pose a global health concern. Resistance mechanisms, including genetic mutations in antimicrobial target genes, efflux pumps, and drug deactivating enzymes, hinder clinical treatment. These resistance factors often spread through mobile genetic elements. Molecular techniques like whole genome sequencing (WGS), multilocus sequence typing (MLST), and phylotyping help decode bacterial genomes and categorise resistance genes.In this study, we analysed 57 UPEC isolates from different UTI patients following EUCAST guidelines. A selection of 17 representative strains underwent WGS, phylotyping, MLST, and comparative analysis to connect laboratory susceptibility data with predictive genomics based on key resistance genes and chromosomal mutations in antimicrobial targets.Trimethoprim resistance consistently correlated with dfr genes, with six different alleles detected among the isolates. These dfr genes often coexisted with class 1 integrons, with the most common gene cassette combining dfr and aadA. Furthermore, 52.9% of isolates harboured the blaTem-1 gene, rendering resistance to ampicillin and amoxicillin. Ciprofloxacin-resistant strains exhibited mutations in GyrA, GyrB and ParC, plasmid-mediated quinolone resistance genes (qnrb10), and aac(6′)-Ib-cr5. Nitrofurantoin resistance in one isolate stemmed from a four amino acid deletion in NfsB.These findings illustrate the varied strategies employed by UPEC to resist antibiotics and the correlation between clinical susceptibility testing and molecular determinants. As molecular testing gains prominence in clinical applications, understanding key resistance determinants becomes crucial for accurate susceptibility testing and guiding effective antimicrobial therapy.

Comprehensive analysis of multiple cytokines to stratify uropathogenic Escherichia coli pathogenesis in mouse model of urinary tract infection

PurposeUrinary tract infection (UTI) is one of the most common human bacterial infections primarily caused by uropathogenic E. coli (UPEC). Empiric treatment in UTI cause emergence of multidrug resistance and limit treatment options. Understanding UTI at the molecular level with respect to the causative pathogen as well as subsequent host response pose an absolute necessity towards appropriate clinical management. This study aimed to investigate host cytokine response in mouse UTI model with respect to bacterial colonization and associated virulence gene expression upon infection. MethodMouse UTI model was established with two clinical UPEC isolates E. coli NP105 and E. coli P025. UPEC colonization in bladder and kidney was evaluated by bacterial culture (CFU/ml). Histopathology of the tissues were examined by hematoxylin and eosin staining. PCR and real time PCR were used to detect the incidence and expression of respective bacterial genes. Cytokine concentrations in tissues and sera were evaluated using ELISA. GraphPad prism version 8.0.2 was used for statistical interpretation. ResultHighest bacterial colonization was observed on 7th and 9th day post infection (p.i). in bladder and kidney of mouse infected with E. coli P025 and E. coli NP105 respectively with a distinct difference in relative expression of fimH and papC adhesin genes in vivo. IL-1β level in tissues and sera of E. coli NP105 and E. coli P025 infected mouse was significantly different but the IL-17A, GCSF, TGF-β levels were comparable. ConclusionThese findings show a role of IL1β to stratify pathogenicity of UPEC in mouse UTI model.

Biofilm formation and antimicrobial resistance pattern of uropathogenic E. coli ST131 isolated from children with malignant tumors

The multidrug-resistant clone identified as Escherichia coli sequence type 131 (E. coli ST131) has spread world-wide. This study sought to ascertain the frequency and biofilm formation of E. coli ST131 isolated from children with various malignancies. A total of 60 uropathogenic E. coli (UPEC) isolates from children without cancer and 30 UPEC isolates from children with cancer were assessed in this study. The microdilution method was used to investigate the sensitivity of bacteria to antibiotics. The microtiter plate (MTP) approach was used to phenotypically assess biofilm formation. The lasR, pelA, and lecA biofilm-encoding genes were detected by PCR in biofilm-producing isolates of E. coli. Thirty-seven out of 90 E. coli isolates were found to be ST131 (41.1%), with 17 (56.7%) from cancer-affected children and 20 (33.3%) from children without cancer, respectively (P-value = 0.036). The frequency of antimicrobial resistance was higher in ST131 strains were compared to non-ST131 strains and when they were isolated from healthy children vs. those who had cancer. In contrast to non-ST131 isolates, ST131 isolates were more biofilm-producers. There was a significant difference between the percentage of biofilm producers between the 22 (100%) ST131-O16 isolates and the 13 (86.7%) ST131-O25b isolates (P-value = 0.04). Children with cancer are more likely than children without cancer to develop biofilm forming E. coli ST131, the latter having a higher profile of antibiotic resistance. Interestingly, E. coli ST131 isolates from non-cancer patients had higher levels of overall antibiotic resistance and while more E. coli ST131isolates from cancer patients formed biofilms.

Molecular characterization of ESBL-producing uropathogenic Escherichia coli isolates among kidney transplant patients: Emergence and spread of B2-ST131 clone type

This study was conducted to identify the distribution of virulence determinants in uropathogenic Escherichia coli (UPEC) isolates obtained from kidney transplant (KTP) and non-transplant patients (non-KTP) with urinary tract infections (UTI). Additionally, the (GTG)5 fingerprinting technique was used to investigate the genetic diversity of Extended-Spectrum B-Lactamase (ESBL)-positive isolates.In this case-control study, 111 urine isolates were obtained from non-KTPs and KTPs, respectively. The presence of genetic markers encoding adhesion proteins, toxins and major E. coli phylogroups was assessed through PCR amplification. Molecular typing of ESBL-positive UPEC strains was performed using (GTG)5 fingerprinting and Multilocus sequence typing (MLST) techniques.Overall, 65 and 46 UPEC isolates were obtained from non-KTPs and KTPs, respectively. Among the studied isolates, traT (85.6%) gene was the most frequently observed virulence gene, followed by kpsMT (49.5%). Using the 80% cut-off point, all the 35 UPEC isolates were classified into four major clusters, namely A, B, C, and D. The majority of the Sequence Type (ST) 131 isolates belonged to cluster A. Additionally, three ST1193 isolates belonged to cluster A and phylogroup B2. Moreover, ST38, ST131 and ST10 were in different cluster.In general, we observed significant differences in the papA, ompT, sat, and vat genes between KTPs and non-KTPs. Furthermore, since all the isolates carried one or more virulence factors (VFs), these findings are concerning in the context of managing UTIs caused by the UPEC strain. Additionally, the distribution of ST and Clonal Complex (CC) among isolates in the main clusters revealed significant differences between MLST and (GTG)5 fingerprinting analysis.

Detection of phylogrouping, adhesin, and extended spectrum β-lactamases genes in hospital acquired uropathogenic Escherichia coli isolates.

It has been interesting to compare the levels of antimicrobial resistance and the virulence characteristics of uropathogenic Escherichia coli (UPEC) strains of certain phylogenetic groups. The purpose of this study was to identify the frequency of phylogenetic groups, adhesin genes, antibiotic sensitivity patterns, and extended spectrum-lactamases (ESBLs) genes in hospital-acquired UPEC. After UPEC isolation, the disc diffusion method was used to assess its susceptibility to antibiotics. Combination disc testing confirmed the existence of ESBL producers. Polymerase chain reaction (PCR) was used to detect genes for adhesin and ESBLs. One hundred and twenty-eight E. coli were isolated which had the highest resistance to tetracycline (96%) followed by cefoxitin (93%), cefepime (92%), ceftazidime (79%), aztreonam (77%) and sulfamethoxazole-trimethoprim (75%). About 57% of isolates were phenotypically ESBLs positive and they were confirmed by PCR. B2 phylogroup (41%) was the most frequent in E. coli isolates then group D (30%), group A (18%), and lastly group B1 (11%). ESBLs genes were more significantly prevalent in phylogroups B2 and D than other phylogroups (P < 0.001). Regarding adhesin genes, both fim H and afa were more significantly associated with group B2 than other groups (P < 0.009, < 0.032), respectively. In ESBL-positive isolates, both genes were more significantly detected compared to negative ones (P < 0.001). Phylogroups B2 and D of UPEC are important reservoirs of antimicrobial resistance and adhesion genes. Detection of ESBL-producing E. coli is important for appropriate treatment as well as for effective infection control in hospitals.

The Efficacy of Silver Nanoparticles in Combating Biofilm Formation by Uropathogenic Escherichia coli

Uropathogenic Escherichia coli (UPEC) is prevalent in urinary tract infections (UTIs). UPEC’s biofilm production enables it to invade and persist in the uroepithelium, leading to recurrent UTIs. The biofilm formation is associated with antibiotic resistance. To overcome this resistance, non-conventional compounds must be developed as an alternative to conventional antibiotics. Silver nanoparticles (AgNPs) are significant due to their antibacterial activity against diverse organisms. This study was done to investigate the antibacterial and anti-biofilm effects of AgNPs on UPEC. AgNPs were biosynthesized using Pseudomonas aeruginosa ATCC 27853. AgNPs were characterized using visual inspection and scanning electron microscopy. The Agar well diffusion method was employed to assess the antibacterial activity of AgNPs against UPEC isolates. The study utilized the tissue culture plate method to investigate both the biofilm and anti-biofilm properties of AgNPs. Following incubation, Ps.aeruginosa and silver nitrate (AgNO3) mixture exhibited a colour change from pale yellow to dark brown. The mean size of spherical AgNPs observed under a scanning electron microscope was 24.187 ± 8.019 nm. 130 UPECs were obtained. AgNPs exhibited antibacterial activity at a concentration of 20 µg/ml against all tested UPEC strains. Among UPEC strains that produced biofilms, a significant inhibition of 99.89 ± 0.45% was observed at a higher concentration of 512 µg/ml of AgNPs. Ps.aeruginosa produces nitrate reductase enzyme that can potentially convert AgNO3 to AgNPs. The biosynthesized AgNPs exhibit antibacterial and anti-biofilm activity against all tested UPEC strains.

Synergistic Antimicrobial Activity of Eugenol in Combination with Fosfomycin to Combat Escherichia coli and Potential Effect on Plasmid-Mediated Fosfomycin Resistance Genes.

The presence of multidrug-resistant pathogenic microorganisms makes it challenging to cure bacterial illnesses. Syzygium aromaticum has been used for medicinal purposes since ancient times. The objective of this study was to investigate the potential synergistic effect of the combination of Eugenol and Fosfomycin against clinically Uropathogenic Escherichia coli (UPEC) and their possible co-treatment as well as their contribution to plasmid-mediated Fosfomycin resistance (fosA3 and fosA4) genes using molecular assays. Eugenol was extracted from clove (Syzygium aromaticum) plants using steam distillation by Clevenger and analyzed by high-performance liquid chromatography (HPLC). UPEC accounted for 63.6 % of all isolates. Specifically, 99.3 % of the UPEC isolates exhibited resistance to multiple types of antibiotics [multidrug-resistant (MDR)]. The MIC for Eugenol was 1.25-5 μg/mL, and Fosfomycin was 512-1024 μg/mL, while the MBC for Eugenol was 5-10 μg/mL and Fosfomycin was 2048 μg/mL. The synergistic effects were considerable, with 1/4 MIC of Eugenol resulting in 1/8 MIC Fosfomycin. Eugenol inhibited most of the UPEC isolates at 4-8 hours, Fosfomycin at 8-12 hours, and co-treatment at 4-8 hours. The fosA3 and fosA4 genes were detected in 5.7 % and 2.9 % of the isolates, respectively. The results showed variable gene expression changes in response to the different treatments.

Repurposing carvacrol, cinnamaldehyde, and eugenol as potential anti-quorum sensing agents against uropathogenic Escherichia coli isolates in Alexandria, Egypt

BackgroundUrinary tract infections represent one of the most frequent hospital and community-acquired infections with uropathogenic Escherichia coli (UPEC) being the main causative agent. The global increase in the emergence of multidrug-resistant (MDR) UPEC necessitates exploring novel approaches. Repurposing natural products as anti-quorum sensing (QS) agents to impede bacterial virulence is gaining momentum nowadays. Hence, this study investigates the anti-QS potentials of carvacrol, cinnamaldehyde, and eugenol against E. coli isolated from urine cultures of Egyptian patients.ResultsAntibiotic susceptibility testing was performed for 67 E. coli isolates and 94% of the isolates showed MDR phenotype. The usp gene was detected using PCR and accordingly, 45% of the isolates were categorized as UPEC. Phytochemicals, at their sub-inhibitory concentrations, inhibited the swimming and twitching motilities of UPEC isolates, with eugenol showing the highest inhibitory effect. The agents hindered the biofilm-forming ability of the tested isolates, at two temperature sets, 37 and 30 °C, where eugenol succeeded in significantly inhibiting the biofilm formation by > 50% at both investigated temperatures, as compared with untreated controls. The phytochemicals were shown to downregulate the expression of the QS gene (luxS) and critical genes related to motility, asserting their anti-QS potential. Further, the combinatory activity of the phytoproducts with five antibiotics was assessed by checkerboard assay. The addition of the phytoproducts significantly reduced the minimum inhibitory concentrations of the antibiotics and generated several synergistic or partially synergistic combinations, some of which have not been previously explored.ConclusionsOverall, carvacrol, cinnamaldehyde, and eugenol could be repurposed as potential anti-QS agents, which preferentially reduce the QS-based communication and attenuate the cascades of gene expression, thus decreasing the production of virulence factors in UPEC, and eventually, subsiding their pathogenicity. Furthermore, the synergistic combinations of these agents with antibiotics might provide a new perspective to circumvent the side effects brought about by high antibiotic doses, thereby paving the way for overcoming antibiotic resistance.

Recent Data Characterizing the Prevalence and Resistance Patterns of FimH-producing Uropathogenic Escherichia coli Isolated from Patients with Urinary Tract Infections in North Lebanon

Background: The adhesin gene (FimH) of uropathogenic Escherichia coli (UPEC) plays a critical role in mediating the first contact of UPEC bacterial strains with uroepithelial cells, leading to colonization and invasion of host cells. Objectives: This study aimed to determine the prevalence of FimH in UPEC strains isolated from patients with urinary tract infections (UTIs) in North Lebanon and characterize the resistance profile of UPEC isolates. Methods: A total of 881 urine samples were collected from UTI-symptomatic patients admitted to different hospitals and laboratories in North Lebanon. Seventy UPEC isolates were identified and transferred to the Biomedical Laboratory of Beirut Arab University (BAU) for further analysis. All UPEC isolates were subjected to antimicrobial susceptibility testing, phenotypic assays for ESBL detection, and PCR to detect the FimH gene. Results: The prevalence of UTIs reached 42% (370/881), with UPEC representing 19% (70/370) of the detected uropathogens. The highest and the lowest resistance among UPEC isolates were reported against Ampicillin (80%; 56/70) and carbapenem (0%; 0/70), respectively. A high prevalence of MDR (68%; 48/70) and ESBL (64%; 45/70) was reported. Molecular analysis revealed that most of the tested UPEC (98.6%; 69/70) harbored the FimH gene. A significant correlation was found between FimH and the antimicrobial resistance properties of UPEC (P &lt; 0.05). Conclusions: This study highlighted the high prevalence of the FimH adhesin gene among UPEC isolates, revealing its crucial role in enhancing the resistance of these bacteria to antimicrobial agents.

Relationship between antimicrobial resistance and virulence factors in uropathogenic Escherichia coli isolates from Ramadi, Iraq: phenotype and genotype identification.

Uropathogenic Escherichia coli (UPEC) is the most predominant pathogen that causes severe urinary tract infections (UTIs). Their therapeutic options are limited due to the rising of antibiotic resistance. The aim of the study was to evaluate the level of antibiotic resistance profile, redundancy of virulence genes, and their correlation. 41 UPEC isolates were collected from patients diagnosed with UTI, identified by the standard microbiological analysis, and tested for susceptibility to 12 antibiotic agents using the Kirby-Bauer method. The ability of UPEC isolates to produce biofilm, hemolyze and cause clumping of blood was determined. Virulence genes were detected by PCR analysis. The percentage of UPEC isolates was higher in females (78.1%) than in males (21.9%). UPEC isolates showed a high degree of resistance towards Ceftriaxone (90.2%), Cefepime (90.2%), Ciprofloxacin (82.9%), Levofloxacin (82.9%), and Trimethoprim-Sulfamethoxazole (80.4%). Biofilm formation (87.8%) and hemagglutinin activity (80.4%) were the most predominant virulence markers expressed in UPEC and showed a high degree of correlation with the antibiotic resistance profile. PCR analysis showed that fimH (85.3%) was the most prevalent gene detected in UPEC isolates, followed by aac3-II (80.4%) among the five genes tested, blaTEM, aac3-II, sul2, hlyA, and fimH. The correlation between antibiotic resistant patterns and the presence aac3-II gene was significantly high. The resistance to the sulfonamides' combined antibiotic was highly correlated with the presence of sulf2 gene. Antimicrobial resistance was significantly linked to phenotypic and genotypic virulence factors. These results will aid in elucidating the pathogenicity of UTIs and guiding treatment decisions.

Quinolone resistance and biofilm formation capability of uropathogenic Escherichia coli isolates from an Iranian inpatients' population.

Uropathogenic Escherichia coli (UPEC) is a major pathogen of the urinary tract infection (UTI), and biofilm formation is crucial as it facilitates the colonization in the urinary tract. We aimed to investigate the antibiotic susceptibility pattern, biofilm formation capability, distribution of quinolone resistance genes, and phylogenetic groups among UPEC isolates from an Iranian inpatients' community. A collection of 126 UPEC obtained from hospitalized patients with symptomatic UTI at 3 teaching hospitals during 2016 were included. Antibiogram of all isolates against quinolone and fluoroquinolones was performed using the disk diffusion method. Phylogenetic groups and qnr A, B, and S genes were assessed by PCR. Susceptibility pattern showed that more than 50% and 81% of the isolates were resistant to fluoroquinolones and quinolones, correspondingly. The frequency of qnrS and qnrB genes was 22% and 13.5%, correspondingly. Our result indicated no significant association between the presence of fluoroquinolone genes and antibiotic resistance to them. The frequent common phylogroup was B2 (84.1%), followed by D (10.3%), A (3.2%) and B1 (2.4%) groups. Indeed, 80.2% of the isolates were biofilm producers, so that 42.1%, 16.7% and 21.4% of them were classified as weak, moderate and strong producers, respectively. Our results showed considerable fluoroquinolone and quinolone resistance among UPEC along with a remarkable rate of biofilm-producing isolates from symptomatic hospitalized patients, making them a serious health concern in the region. This survey highlights the need for awareness on quinolone resistance and careful prescription of them by physicians.

High incidence of fosfomycin-resistant uropathogenic E. coli among children

BackgroundThere are few epidemiological or molecular data on Escherichia coli (E. coli) strains resistant to fosfomycin. In this study, we described the occurrence and characterization of fosfomycin-resistant uropathogenic E. coli (UPEC) isolated from children.Materials and methodsThis study was carried out on 96 E. coli isolates obtained from children with urinary tract infections. Two methods were performed to detect fosfomycin resistance: The agar dilution method and the rapid fosfomycin test. The disc diffusion method was done to detect the antimicrobial susceptibility pattern of all isolates. The phylogenetic grouping of all isolates was done according to the modified Clermont method. Conventional PCR was performed to detect plasmid-mediated fosfomycin-resistant genes (fos genes) and the blaCTX−M gene.ResultsAnalyses of data were performed by SPSS software. A high percentage of fosfomycin resistance (37/96; 38.5%) was reported among UPEC isolates. The fosfomycin-resistant strains showed a higher resistance rate than fosfomycin-susceptible isolates to different antibiotics. E group (62.2%) was the most predominant phylogenetic group among the fosfomycin-resistant UPEC isolates, followed by Group B2 (21.6%) and group D (13.5%). The fos genes were detected in 21 isolates with the fosA3 gene as the most frequent, which was detected in 11 isolates followed by fosA (8), fosC2 (4), fosA4(1), and fosA5(1) genes.ConclusionThis is the first report of a high prevalence of plasmid-mediated fosfomycin-resistant UPEC in Egypt. All of these isolates were multidrug-resistant to the tested antibiotics. Close monitoring of such strains is mandatory to prevent widespread dissemination of the genes code for antibiotic resistance.

Contribution of Antibiotic Susceptibility Testing and CH Typing Compared to Next-Generation Sequencing for the Diagnosis of Recurrent Urinary Tract Infections Due to Genetically Identical Escherichia coli Isolates: a Prospective Cohort Study of Cystitis in Women.

Recurrent cystitis is a common disease in women, mainly due to uropathogenic Escherichia coli (UPEC). For decades, typing methods now considered obsolete suggested that relapse by the same clone is dominant over reinfection, most UPEC strains being otherwise fully susceptible to antibiotics. We aimed to update these data. Thanks to a prospective study over 17 months, we recruited 323 women with cystitis. Of these, 251 of them had sporadic infection and 72 had recurrence, with 2 to 9 episodes per patient for a total of 131 UPEC isolates and 145 UPEC pairs at patient level. Phylogroups B2 (52.4%) and D (14.1%) were overall dominant, as expected due to their particular urovirulence. CH typing identified 119 distinct profiles with no CH type particularly associated with recurrence. Relapse was attested by CH typing for only 30.6% (22 out of 72), with very diverse situations ranging from all episodes due to the same clone to alternating reinfections and relapses. Next-generation sequencing confirmed the clonality for all but two of the 145 UPEC pairs. Antibiotic resistance was common for recurrent cystitis isolates (only 25 [17.2%] out of 145 UPEC pairs were fully susceptible), allowing us to predict UPEC clonality. Indeed, antibiotic susceptibility profile matched CH typing for 104 (71.7%) pairs. Finally, we demonstrated a large genetic diversity among UPEC isolates responsible for cystitis in women, even in cases of recurrence for which reinfection appeared dominant over relapse. Recurrent cystitis appears to be a heterogeneous disease requiring tailored treatment and prevention. IMPORTANCE More than half of women will experience cystitis during their lifetime. Among these women, 25% will experience a second episode within the following 6 months. It is epidemiologically important to discriminate relapses from reinfections. Relapse identification relies on long and laborious methods and might influence treatment. Therefore, the designation of time- and cost-effective strategies for this goal is of particular interest. Our work suggests using CH typing and antibiotic susceptibility profiles to type Escherichia coli, the main uropathogen.

Presence of fimH and iss type 1, 2 and 3 genes in uropathogenic Escherichia coli isolates recovered from an apex medical institute in North India

PurposeTo detect the presence of fimH and iss type 1, 2 and 3 genes in uropathogenic Escherichia coli (UPEC) isolates recovered from patients coming to the out patient department (OPD) of our hospital. MethodsE. coli isolates recovered from patients who had symptoms of urinary tract infection (UTI) were processed for the presence of fimH and iss genes. DNA was extracted using an in house method after which conventional PCR using forward and reverse primers targeting the four genes was carried out. The amplified products were electrophoresed and visualized in a gel documentation imager. Relevant demographic details of the patients were recorded on a pre-designed pro-forma and antimicrobial susceptibility testing of the isolates was done by disc diffusion method. ResultsfimH was present in 87.5% of UPEC isolates whereas iss type 1 was seen in 7.3%, type 2 in 4.2% and iss type 3 in 71.9% isolates. Age of the patients ranged from 3 months to 82 ​yrs (mean 43.5 SD ​± ​18.20). UTI was more common in females (60.2%) as compared to males patients (39.8%). Dysuria (66.7%) was the most common symptom in the studied subjects and diabetes mellitus (42.6%) the most common co-morbidity. A total of 56.5% patients gave a history of prior antibiotic intake. The UPEC isolates were resistant to most of the antibiotics tested. However all the isolates were sensitive to polymyxin B and colistin. Fosfomycin resistance was seen in 9.5% of the UPEC isolates harbouring fimH gene. ConclusionThis is the first study that highlights the presence of iss type 3 gene in UPEC isolates along with the fimH and iss type 1 and 2 genes. The results of this study can serve as a stepping stone for future in depth research into the significance of the iss genes in causing UTI.

The use of Escherichia coli strain Nissle 1917 shows promise for improving gastrointestinal and urinary health in dogs.

To investigate the probiotic Escherichia coli Nissle 1917 (EcN) in canine idiopathic diarrhea and urinary tract infections. The utility of EcN was explored in a 3-phase study from March 2017 to June 2020. Eighty-nine dogs with idiopathic diarrhea were included in phase 1, 3 healthy dogs were included in phase 2, and uropathogenic E coli (UPEC) isolates from 38 dogs with urinary tract infections were included in phase 3. In phase 1, dogs with diarrhea were prospectively enrolled in a randomized study to receive EcN (108 EcN bacteria/mL; < 10 kg received 5 mL/dose, 10 to 25 kg received 10 mL/dose, or > 25 kg received 15 mL/dose) or placebo for 3 days, followed by a 15-day observation phase. In phase 2, healthy dogs received EcN as described in phase 1, with feces analyzed for E coli populations and microbiome composition at days 0, 3, and 7. In phase 3, EcN efficacy was tested by in vitro plate assay against UPEC isolates. Median duration of abnormal stool consistency, time to response, and duration of diarrhea were shorter for dogs that received EcN (5.0, 3.0, and 2.0 days, respectively) versus the placebo (7.0, 5.0, and 4.0 days, respectively) (P = .21, P = .05, and P = .039, respectively). EcN induced shifts in E coli diversity in healthy dogs while having minimal impact on overall microbiome structure. Furthermore, 68% of the canine UPEC isolates were susceptible to EcN in vitro. EcN improved the treatment of idiopathic diarrhea, colonized the gastrointestinal tract during the trial, and displayed in vitro competition with UPEC.

Genotypic characteristics of Uropathogenic Escherichia coli isolated from complicated urinary tract infection (cUTI) and asymptomatic bacteriuria-a relational analysis.

Uropathogenic Escherichia coli (UPEC) is the predominant agent causing various categories of complicated urinary tract infections (cUTI). Although existing data reveals that UPEC harboured numerous virulence determinants to aid its survival in the urinary tract, the reason behind the occurrence of differences in the clinical severity of uninary tract infections (UTI) demonstrated by the UPEC infection is poorly understood. Therefore, the present study aims to determine the distribution of virulence determinants and antimicrobial resistance among different phylogroups of UPEC isolated from various clinical categories of cUTI and asymptomatic bacteriuria (ASB) E. coli isolates. The study will also attempt a relational analysis of the genotypic characteristics of cUTI UPEC and ASB E. coli isolates. A total of 141 UPEC isolates from cUTI and 160 ASB E. coli isolates were obtained from Universiti Malaya Medical Centre (UMMC). Phylogrouping and the occurrence of virulence genes were investigated using polymerase chain reaction (PCR). Antimicrobial susceptibility of the isolates to different classes of antibiotics was determined using the Kirby Bauer Disc Diffusion method. The cUTI isolates were distributed differentially among both Extraintestinal Pathogenic E. coli (ExPEC) and non-ExPEC phylogroups. Phylogroup B2 isolates were observed to possess the highest average aggregative virulence score (7.17), a probable representation of the capability to cause severe disease. Approximately 50% of the cUTI isolates tested in this study were multidrug resistant against common antibiotics used to treat UTI. Analysis of the occurrence of virulence genes among different cUTI categories demonstrated that UPEC isolates of pyelonephritis and urosepsis were highly virulent and had the highest average aggregative virulence scores of 7.80 and 6.89 respectively, compared to other clinical categories. Relational analysis of the occurrence of phylogroups and virulence determinants of UPEC and ASB E. coli isolates showed that 46.1% of UPEC and 34.3% of ASB E. coli from both categories were distributed in phylogroup B2 and had the highest average aggregative virulence score of 7.17 and 5.37, respectively. The data suggest that UPEC isolates which carry virulence genes from all four virulence genes groups studied (adhesions, iron uptake systems, toxins and capsule synthesis) and isolates from phylogroup B2 specifically could predispose to severe UTI involving the upper urinary tract. Therefore, specific analysis of the genotypic characteristics of UPEC could be further explored by incorporating the combination of virulence genes as a prognostic marker for predicting disease severity, in an attempt to propose a more evidence driven treatment decision-making for all UTI patients. This will go a long way in enhancing favourable therapeutic outcomes and reducing the antimicrobial resistance burden among UTI patients.

Study of antibiotic resistance and virulence genes in Escherichia coli isolated from urinary tract infection patients in Tanta University Hospitals.

This study aimed to investigate the prevalence of antibiotic resistance genes CTX-M and Qnr, as well as the virulence genes HlyA, Pap, CNF1, and Afa, in uropathogenic Escherichia coli (UPEC) isolates from the Egyptian population. In this cross-sectional study, a total of 50 E. coli isolates were collected from urine samples from patients with urinary tract infections (UTIs) admitted to Tanta University Hospital from December 2020 to November 2021. The isolates were cultured, identified, and tested for antibiotic susceptibility by the disc diffusion method. The CTX-M, Qnr (QnrA, B, and S), Pap, CNF1, HlyA, and Afa genes were detected by polymerase chain reaction in UPEC isolates. The Pap, CNF1, HlyA, and Afa genes were found to be positive in 18%, 12%, 10%, and 2% of the isolates, respectively. In addition, CTX-M and QnrS were found to be positive in 44% and 8% of the isolates, while QnrA and B were not detected. Furthermore, positive Pap, CNF1, and HlyA genes were significantly associated with both upper and lower UTIs, increased frequency, urgency, and dysuria, and complicated UTIs, as well as pyuria over 100 white blood cells per high-power field. In conclusion, the prevalence of virulence and antibiotic resistance genes varies by population. At our hospital, the Pap gene is the most prevalent virulence gene and was strongly associated with complicated UTIs, while the CTX-M and QnrS genes were the most prevalent and related to antibiotic resistance. Our findings, however, should be interpreted with caution due to the small sample size.

Antimicrobial resistance and molecular epidemiology of carbapenem-resistant Escherichia coli from urinary tract infections in Shandong, China.

Urinary tract infection (UTI) is one of the most common extraintestinal infections, and uropathogenic Escherichia coli (UPEC) is the main cause of UTIs. However, the ability to treat UTI has been compromised by the increase in antimicrobial resistance, especially carbapenem resistance. Here, we aimed to characterize the antimicrobial resistance and molecular epidemiology of carbapenem-resistant UPEC isolated in Shandong, China. In total, 17 carbapenem-resistant UPEC (CR-UPEC) isolates were collected from July 2017 to May 2020 in the Shandong Provincial Hospital. Whole-genome sequencing and bioinformatics analyses were performed to understand the molecular epidemiology of CR-UPEC. Phylogenetic groups, drug resistance genes, biofilm formation, and virulence-related gene profiles of the isolates were analyzed. Plasmid profiling and conjugation assay were performed to evaluate the ability to transfer carbapenem resistance-related genes to other E. coli isolates. Biofilm formation was also evaluated, as it is important for the persistence of infectious diseases. We observed that 15 out of 17 CR-UPEC strains were blaNDM producers, among which 4 isolates could transfer blaNDM to recipient cells. The predominant sequence type was ST167 (6/17), followed by ST410 (3/17). The most prevalent phylogenetic group was phylogenetic group A (10/17), followed by phylogenetic group C (3/17). One isolate was resistant to polymyxin, which was caused by the carriage of a transferable plasmid harboring mcr-1. Statistical analysis did not reveal any significant difference in the carriage rate of fimbriae-coding genes between strong and weak biofilm producers. Our observations may assist in developing new therapeutic methods for drug-resistant organisms.

Genomic Characterization of Uropathogenic Escherichia coli Isolates from Tertiary Hospitals in Riyadh, Saudi Arabia.

Uropathogenic Escherichia coli (UPEC) is the most common cause of urinary tract infections (UTIs) in hospitalised and non-hospitalised patients. Genomic analysis was used to gain further insight into the molecular characteristics of UPEC isolates from Saudi Arabia. A total of 165 isolates were collected from patients with UTIs between May 2019 and September 2020 from two tertiary hospitals in Riyadh, Saudi Arabia. Identification and antimicrobial susceptibility testing (AST) were performed using the VITEK system. Extended-spectrum β-lactamase (ESBL)-producing isolates (n = 48) were selected for whole genome sequencing (WGS) analysis. In silico analysis revealed that the most common sequence types detected were ST131 (39.6%), ST1193 (12.5%), ST73 (10.4%), and ST10 (8.3%). Our finding showed that blaCTX-M-15 gene was detected in the majority of ESBL isolates (79.2%), followed by blaCTX-M-27 (12.5%) and blaCTX-M-8 (2.1%). ST131 carried blaCTX-M-15 or blaCTX-M-27, and all ST73 and ST1193 carried blaCTX-M-15. The relatively high proportion of ST1193 in this study was notable as a newly emerged lineage in the region, which warrants further monitoring.