In vitro-antibacterial properties of ten medicinal plants against common uropathogenic organisms and toxicity determination using brine shrimp lethality assay.

BACKGROUND: In recent years, antibiotic resistance has emerged as a global health concern in bacterial infections such as urinary tract infections (UTIs). Uropathogenic Escherichia coli is the most frequent organism responsible for both simple and complex UTIs. Staphylococcus aureus and Pseudomonas aeruginosa are frequently associated with complicated UTIs. Sri Lanka has significant resources of medicinal plants used to cure UTIs in Ayurvedic and traditional medicine. METHODS: Agar well diffusion and broth microdilution methods were used to determine the antibacterial activity of the methanolic extract of ten medicinal plants against P. aeruginosa ATCC27853, S.aureus ATCC25923, E.coli ATCC25922 and their UTI positive strains extracted from positive culture plates. As a preliminary toxicity assay, the Brine Shrimp Lethality Assay (BSLA) was used to determine its cytotoxicity. RESULTS: The methanolic fruits extract of P. emblica demonstrated the highest antibacterial activity against both E. coli ATCC25922 and E. coli UTI-positive strains. B. diffusa roots extract exhibited the highest activity against S. aureus ATCC25923, while T. chebula fruits extract showed the highest activity against the S. aureus UTI-positive strain. T. involucrata roots extract displayed the highest activity against P. aeruginosa ATCC27853, and Z. officinale rhizomes extract showed the highest activity against the P. aeruginosa UTI-positive strain. Moreover, the plant mixture showed the most substantial antibacterial effect against P. aeruginosa ATCC27853. However, the methanolic seed extract of C. melo did not exhibit any antimicrobial effects against the selected organisms. All plant material, including the plant mixture, showed cytotoxicity according to the BSLA. CONCLUSION: All the methanolic extracts including P. emblica fruits, O. tenuiflorum whole plant, T. chebula fruits, Z. officinale rhizome, T. terrestris roots, T. involucrata roots, A. lanata whole plant. B. diffusa roots and A. falcatus roots showed antimicrobial effects against selected strains except C. melo seed extract. The results of the present study evidently supports the traditional and ayurvedic use of these plants for the treatment of UTIs. This paves the way for another praise for new plant-based therapeutic product development for the treatment of UTIs. However, further toxicity studies are needed for medicinal dose determination.

Risk Factors and Mortality Outcomes in Elderly Patients With Bloodstream Infections: A Retrospective Analysis.

Background The objective of our investigation was to evaluate the mortality rate and predictor factors that are associated with bloodstream infections (BSIs) in elderly patients who are admitted to the internal medicine ward. Materials and methods A retrospective cross-sectional analysis was conducted at a 550-bed tertiary care hospital in Peshawar, Pakistan, from January 2021 to June 2022. The study involved elderly inpatients aged 65 and older with positive culture results detected within two days of admission. Data collection involved demographic and patient-related risk variables, BSI-related risk factors, and environmental risk factors, with statistical analysis performed using the Statistical Package for the Social Sciences (IBM SPSS Statistics for Windows, IBM Corp., Version 26.0, Armonk, NY). Results Of the total study sample (n=186), 103 (55.4%) survived while 83 (44.6%) did not. The non-survivor group had a higher median Sequential Organ Failure Assessment (SOFA) score (6 vs. 2, p<0.0001) and Charlson Comorbidity Index (5 ± 2 vs. 3 ± 2, p<0.0001), with more frequent immunosuppression (25.3% vs. 8.7%, p=0.001). Additionally, gram-positive bacteria were more common in non-survivors (42% vs. 10%, p<0.0001), while gram-negative bacteria were more prevalent in survivors (73% vs. 36%, p=0.002). Conclusions Our research validates that BSI in older adults is a serious condition that is linked to a substantial death rate during hospitalization. The biggest determinant of death in older patients with BSI is the severity of clinical symptoms evaluated by the SOFA score upon admission. It is imperative to acknowledge that respiratory-induced BSIs are the most fatal, and patients who are hospitalized and admitted to the intensive care unit (ICU) are at an elevated risk.

Green stabilization of silver nanoparticles over the surface of biocompatible Fe3O4@CMC for bactericidal applications.

The emergence of antimicrobial resistance in bacteria, especially in agents associated with urinary tract infections (UTIs), has initiated an exciting effort to develop biocompatible nanoparticles to confront their threat. Designing simple, cheap, biocompatible, and efficient nanomaterials as bactericidal agents seems to be a judicious response to this problem. Here, a solvothermal method was hired for the one-pot preparation of the cellulose gum (carboxymethyl cellulose, CMC) magnetic composite to prepare a cost-effective, efficient, and biocompatible support for the plant-based stabilization of the silver NPs. The green stabilization of the Ag NPs is performed using Euphorbia plant extract with high efficiency. Various characterization methods, including FT-IR, XRD, SEM, EDS, TEM, and VSM were used to study the composition and properties of Fe3O4@CMC/AgNPs. The composite shows well integrity and monodispersity with a mean diameter of <300 nm, indicating its potential for bio-related application. The CMC functionalities of the proposed material facilitated the stabilization of the Ag NPs, resulting in their monodispersity and enhanced performance. The manufactured composite was used as an antibacterial agent for the removal of UTIs agents, collected from 200 hospitalized patients with acute coronary syndrome, which showed promising results. This study showed that the concentration of the Ag NPs has a direct relationship with the antibacterial properties of the composite.

Isolation and characterization of bacteriophages against E.coli urinary tract infection and evaluating their anti-biofilm activity and antibiotic synergy.

Urinary tract infections (UTIs) by Uropathogenic Escherichia coli (UPEC) are a significant health concern, especially due to the increasing prevalence of antibiotic resistance. This study focuses on isolating and characterizing bacteriophages specific to UPEC strains isolated from UTI samples. The isolated phages were assessed for their ability to target and lyse UPEC in vitro, focusing on their efficacy in disrupting biofilms, a key virulence factor contributing to UTI recurrence and antibiotic resistance. The morphological structure observed by TEM belongs to Myoviridae, the phage exhibited icosahedral symmetry with a long non-constricting tail, the approximate measurement of the phage head was 39 nm in diameter, and the phage tail was 105.317 nm in length. One-step growth experiments showed that the latent period was approximately 20 min, followed by a rise period of 40 min, and a growth plateau was reached within 20 min and the burst size observed was 26 phages/infected bacterial cells. These phages were capable of killing cells within the biofilms, leading to a reduction in living cell counts after a single treatment. This study highlights the potential of phages to play a significant role in disrupting, inactivating, and destroying Uropathogenic Escherichia coli (UPEC) biofilms. Such findings could be instrumental in developing treatment strategies that complement antibiotics and disinfectants. The phage-antibiotic synergistic activity was compared to have the possibility to facilitate the advancement of focused and enduring alternatives to traditional antibiotic therapies for UTIs.

Mendelian randomization analysis reveals higher whole body water mass may increase risk of bacterial infections.

BACKGROUND AND PURPOSE: The association of water loading with several infections remains unclear. Observational studies are hard to investigate definitively due to potential confounders. In this study, we employed Mendelian randomization (MR) analysis to assess the association between genetically predicted whole body water mass (BWM) and several infections. METHODS: BWM levels were predicted among 331,315 Europeans in UK Biobank using 418 SNPs associated with BWM. For outcomes, we used genome-wide association data from the UK Biobank and FinnGen consortium, including sepsis, pneumonia, intestinal infections, urinary tract infections (UTIs) and skin and soft tissue infections (SSTIs). Inverse-variance weighted MR analyses as well as a series of sensitivity analyses were conducted. RESULTS: Genetic prediction of BWM is associated with an increased risk of sepsis (OR 1.34; 95% CI 1.19 to 1.51; P = 1.57 × 10- 6), pneumonia (OR: 1.17; 95% CI 1.08 to 1.29; P = 3.53 × 10- 4), UTIs (OR: 1.26; 95% CI 1.16 to 1.37; P = 6.29 × 10- 8), and SSTIs (OR: 1.57; 95% CI 1.25 to 1.96; P = 7.35 × 10- 5). In the sepsis and pneumonia subgroup analyses, the relationship between BWM and infection was observed in bacterial but not in viral infections. Suggestive evidence suggests that BWM has an effect on viral intestinal infections (OR: 0.86; 95% CI 0.75 to 0.99; P = 0.03). There is limited evidence of an association between BWM levels and bacteria intestinal infections, and genitourinary tract infection (GUI) in pregnancy. In addition, MR analyses supported the risk of BWM for several edematous diseases. However, multivariable MR analysis shows that the associations of BWM with sepsis, pneumonia, UTIs and SSTIs remains unaffected when accounting for these traits. CONCLUSIONS: In this study, the causal relationship between BWM and infectious diseases was systematically investigated. Further prospective studies are necessary to validate these findings.

Clinical Outcomes in Patients Who Received a One-Time Aminoglycoside Dose for Extended-Spectrum Beta-Lactamase-Producing Enterobacterales or Pseudomonas aeruginosa Cystitis.

The Infectious Diseases Society of America (IDSA) recommends a single dose of an aminoglycoside for uncomplicated cystitis caused by extended-spectrum beta-lactamase (ESBL)-producing Enterobacterales (ESBL-E) and difficult-to-treat Pseudomonas aeruginosa. However, there is very little recent clinical evidence to support this recommendation. The objective of this study was to evaluate the safety and efficacy of a single-dose aminoglycoside for cystitis caused by ESBL-E or Pseudomonas aeruginosa. This was a multicenter, retrospective, cohort study. Patients who received ≥3 days of standard of care were compared to patients who received a one-time dose of an aminoglycoside with or without a short course of effective therapy before. The primary outcome was the rate of relapse defined as requiring escalation of antibiotics or starting new antibiotic therapy within 14 days after the completion of antibiotics. A total of 66 patients were included in this study, with 33 patients in each arm. There were more males and complicated cystitis patients in the standard-of-care group. There was no difference found in the rate of relapse. The length of stay was significantly shorter in the aminoglycoside group (4.5 ± 4.4 days vs. 14.1 ± 10.1 days, p < 0.0001). A one-time dose of an aminoglycoside did not increase the risk of relapse and was associated with a shorter length of stay when used to treat cystitis caused by ESBL-E or Pseudomonas aeruginosa.

Therapies in preclinical and in early clinical development for the treatment of urinary tract infections: from pathogens to therapies.

INTRODUCTION: Urinary tract infections (UTIs) are a prevalent health challenge characterized by the invasion and multiplication of microorganisms in the urinary system. The continuous exploration of novel therapeutic interventions is imperative. Advances in research offer hope for revolutionizing the management of UTIs and improving the overall health outcomes for individuals affected by these infections. AREAS COVERED: This review aimed to provide an overview of existing treatments for UTIs, highlighting their strengths and limitations. Moreover, we explored and analyzed the latest therapeutic modalities under clinical development. Finally, the review offered a picture into the potential implications of these therapies on the future landscape of UTIs treatment, discussing possible advancements and challenges for further research. EXPERT OPINION: Comprehensions into the pathogenesis of UTIs have been gleaned from foundational basic science studies, laying the groundwork for the exploration of novel therapeutic interventions. The primary source of evidence originates predominantly from animal studies conducted on murine models. Nevertheless, the lack of clinical trials interferes the acquisition of robust evidence in humans. The challenges presented by the heterogeneity and virulence of uropathogens add an additional layer of complexity, posing an obstacle that scientists and clinicians are actively grappling with in their pursuit of effective solutions.

Regional variations in antimicrobial susceptibility of community-acquired uropathogenic Escherichia coli in India: Findings of a multicentric study highlighting the importance of local antibiograms.

Objectives: Evidence-based prescribing is essential to optimize patient outcomes in cystitis. This requires knowledge of local antibiotic resistance rates. Diagnostic and Antimicrobial Stewardship (DASH) to Protect Antibiotics (https://dashuti.com/) is a multicentric mentorship program guiding centers in preparing, analyzing and disseminating local antibiograms to promote antimicrobial stewardship in community urinary tract infection. Here, we mapped the susceptibility profile of Escherichia coli from 22 Indian centers. Methods: These centers spanned 10 Indian states and three union territories. Antibiograms for urinary E. coli from the outpatient departments were collated. Standardization was achieved by regional online training; anomalies were resolved via consultation with study experts. Data were collated and analyzed. Results: Nationally, fosfomycin, with 94% susceptibility (inter-center range 83-97%), and nitrofurantoin, with 85% susceptibility (61-97%), retained the widest activity. The susceptibility rates were lower for co-trimoxazole (49%), fluoroquinolones (31%), and oral cephalosporins (26%). The rates for third- and fourth-generation cephalosporins were 46% and 52%, respectively, with 54% (33-58%) extended-spectrum ß-lactamase prevalence. Piperacillin-tazobactam (81%), amikacin (88%), and meropenem (88%) retained better activity; however, one center in Delhi recorded only 42% meropenem susceptibility. Susceptibility rates were mostly higher in South, West, and Northeast India; centers in the heavily populated Gangetic plains, across north and northwest India, had greater resistance. These findings highlight the importance of local antibiograms in guiding appropriate antimicrobial choices. Conclusions: Fosfomycin and nitrofurantoin are the preferred oral empirical choices for uncomplicated E. coli cystitis in India, although elevated resistance in some areas is concerning. Empiric use of fluoroquinolones and third-generation cephalosporins is discouraged, whereas piperacillin/tazobactam and aminoglycosides remain carbapenem-sparing parenteral agents.

Antimicrobial susceptibility patterns of urinary tract infections causing bacterial isolates and associated risk factors among HIV patients in Tigray, Northern Ethiopia.

BACKGROUND: Urinary tract infections, a prevalent global infectious disease, are clinical issues not well studied in HIV-positive individuals. UTIs have become a global drug resistance issue, but the prevalence and antibiotic susceptibility patterns of UTI-causing bacteria among HIV patients in Tigray, Ethiopia, are poorly understood. This study aims to identify the prevalence of UTI-causing bacteria, their antibiotic susceptibility patterns, and associated risk factors in HIV patients attending ART clinics at Mekelle General Hospital and Ayder Comprehensive Specialized Hospital in Tigray, Northern Ethiopia. METHOD: Clean-catch midstream urine samples (10-15 mL) were collected from HIV patients who are attending ART clinics at Mekelle General Hospital and Ayder Comprehensive Specialized Hospital. Samples were analyzed based on standard microbiological protocols using cysteine-lactose electrolyte deficient (CLED) agar. Pure colonies of bacterial isolates were obtained by sub-culturing into Mac-Conkey, Manitol Salt agar and blood agar plates. The bacterial isolates were then identified using macroscopic, microscopic, biochemical, and Gram staining methods. Gram-negative bacteria were identified using biochemical tests like triple sugar iron agar, Simon's citrate agar, lysine iron agar, urea, motility test, and indol test, whereas Gram-positive isolates were identified using catalase and coagulase tests. The Kirby-Bauer disk diffusion technique was used to analyze the antimicrobial susceptibility pattern of bacterial isolates. Data was analyzed using SPSS version 25.0. RESULTS: Among the 224 patients, 28 (12.5%) of them had been infected by UTIs-causing bacteria. E. coli was the dominant bacterium (16 (57%)) followed by K. pneumoniae (4 (14%)), and S. aureus (3 (11%)). Of the total bacterial isolates, 22 (78.6%) of them developed multi-drug resistance. All Gram-positive (100%) and 75% of Gram-negative bacterial isolates were found to be resistant to two or more drugs. Patients with a history of UTIs, and with CD4 count < 200 cells/ mm3, were more likely to have significant bacteriuria. Compared to male patients, female patients were more affected by the UTIs-causing bacteria. More than 93% of the UTIs-causing bacterial isolates were susceptible to nitrofurantoin, ceftriaxone, ciprofloxacin, and gentamycin; whereas they are highly resistant to ampicillin (96%), cotrimoxazole (82%) and tetracycline (71%). CONCLUSIONS: Most of the bacterial isolates were highly resistant to ampicillin, cotrimoxazole, and tetracycline. Female patients were more affected by the UTIs causing bacteria. The highest prevalence (12.5%) of UTIs in HIV patients needs special attention for better management and monitoring. Previous UTI history and immune suppression are predictors of UTIs, highlighting the need for intervention measures involving molecular studies to identify resistant bacteria genes and promote patient immune reconstitution.

ColiSeq: a multiplex amplicon assay that provides strain level resolution of Escherichia coli directly from clinical specimens.

Escherichia coli is a diverse pathogen, causing a range of disease in humans, from self-limiting diarrhea to urinary tract infections (UTIs). Uropathogenic E. coli (UPEC) is the most frequently observed uropathogen in UTIs, a common disease in high-income countries, incurring billions of dollars yearly in treatment costs. Although E. coli is easily grown and identified in the clinical laboratory, genotyping the pathogen is more complicated, yet critical for reducing the incidence of disease. These goals can be achieved through whole-genome sequencing of E. coli isolates, but this approach is relatively slow and typically requires culturing the pathogen in the laboratory. To genotype E. coli rapidly and inexpensively directly from clinical samples, including but not limited to urine, we developed and validated a multiplex amplicon sequencing assay, called ColiSeq. The assay consists of targets designed for E. coli species confirmation, high resolution genotyping, and mixture deconvolution. To demonstrate its utility, we screened the ColiSeq assay against 230 clinical urine samples collected from a hospital system in Flagstaff, Arizona, USA. A limit of detection analysis demonstrated the ability of ColiSeq to identify E. coli at a concentration of ~2 genomic equivalent (GEs)/mL and to generate high-resolution genotyping at a concentration of 1 × 105 GEs/mL. The results of this study suggest that ColiSeq could be a valuable method to understand the source of UPEC strains and guide infection mitigation efforts. As sequence-based diagnostics become accepted in the clinical laboratory, workflows such as ColiSeq will provide actionable information to improve patient outcomes.IMPORTANCEUrinary tract infections (UTIs), caused primarily by Escherichia coli, create an enormous health care burden in the United States and other high-income countries. The early detection of E. coli from clinical samples, including urine, is important to target therapy and prevent further patient complications. Additionally, understanding the source of E. coli exposure will help with future mitigation efforts. In this study, we developed, tested, and validated an amplicon sequencing assay focused on direct detection of E. coli from urine. The resulting sequence data were demonstrated to provide strain level resolution of the pathogen, not only confirming the presence of E. coli, which can focus treatment efforts, but also providing data needed for source attribution and contact tracing. This assay will generate inexpensive, rapid, and reproducible data that can be deployed by public health agencies to track, diagnose, and potentially mitigate future UTIs caused by E. coli.

MANNosylation of Mesoporous Silica Nanoparticles Modifies TLR4 Localization and NF-κB Translocation in T24 Bladder Cancer Cells.

D-mannose is widely used as non-antibiotic treatment for bacterial urinary tract infections. This application is based on a well-studied mechanism of binding to the type 1 bacterial pili and, therefore, blocking bacteria adhesion to the uroepithelial cells. To implement D-mannose into carrier systems, the mechanism of action of the sugar in the bladder environment is also relevant and requires investigation. Herein, two different MANNosylation strategies using mesoporous silica nanoparticles (MSNs) are described. The impact of different chemical linkers on bacterial adhesion and bladder cell response is studied via confocal microscopy imaging of the MSN interactions with the respective organisms. Cytotoxicity is assessed and the expression of Toll-like receptor 4 (TLR4) and caveolin-1 (CAV-1), in the presence or absence of simulated infection with bacterial lipopolysaccharide (LPS), is evaluated using the human urinary bladder cancer cell line T24. Further, localisation of the transcription factor NF-κB due to the MANNosylated materials is examined over time. The results show that MANNosylation modifies bacterial adhesion to the nanomaterials and significantly affects TLR4, caveolin-1, and NF-κB in bladder cells. These elements are essential components of the inflammatory cascade/pathogens response during urinary tract infections. These findings demonstrate that MANNosylation is a versatile tool to design hybrid nanocarriers for targeted biomedical applications.

Characterization and synergy studies of Caudoviricete Escherichia phage FS2B infecting multi-drug resistant uropathogenic Escherichia coli isolates / Estudios de caracterización y sinergia del fago FS2B de Caudoviricete Escherichia que infecta aislados de Escherichia coli uropatógena multirresistente

Escherichia coli is one of the most common causes of urinary tract infections. However, a recent upsurge in antibiotic resistance among uropathogenic E. coli (UPEC) strains has provided an impetus to explore alternative antibacterial compounds to encounter this major issue. In this study, a lytic phage against multi-drug-resistant (MDR) UPEC strains was isolated and characterized. The isolated Escherichia phage FS2B of class Caudoviricetes exhibited high lytic activity, high burst size, and a small adsorption and latent time. The phage also exhibited a broad host range and inactivated 69.8% of the collected clinical, and 64.8% of the identified MDR UPEC strains. Further, whole genome sequencing revealed that the phage was 77,407 bp long, having a dsDNA with 124 coding regions. Annotation studies confirmed that the phage carried all the genes associated with lytic life cycle and all lysogeny related genes were absent in the genome. Further, synergism studies of the phage FS2B with antibiotics demonstrated a positive synergistic association among them. The present study therefore concluded that the phage FS2B possesses an immense potential to serve as a novel candidate for treatment of MDR UPEC strains.(AU)

Two novel phages, Klebsiella phage GADU21 and Escherichia phage GADU22, from the urine samples of patients with urinary tract infection.

Phages are found in a wide variety of places where bacteria exist including body fluids. The aim of the present study was to isolate phages from the urine samples of patients with urinary tract infection. The 10 urine samples were cultured to isolate bacteria and also used as phage sources against the isolated bacteria. From 10 urine samples with positive cultures, 3 phages were isolated (33%) and two of them were further studied. The Klebsiella phage GADU21 and Escherichia phage GADU22 phages infected Klebsiella pneumonia and Escherichia coli, respectively. Among the tested 14 species for host range analysis, the Klebsiella phage GADU21 was able to infect two species which are Klebsiella pneumonia and Proteus mirabilis, and Escherichia phage GADU22 was able to infect four species which are Shigella flexneri, Shigella sonnei and Escherichia coli. Among different isolates of the indicator bacteria for each phage, GADU21 infected half of the tested 20 Klebsiella pneumonia isolates while GADU22 infected 85% of the tested 20 E. coli isolates. The genome sizes and GC ratios were 75,968 bp and 44.4%, and 168,023 bp and 35.3% for GADU21 and GADU22, respectively. GADU21 and GADU22 were both lytic and had no antibiotic resistance and virulence genes. GADU21 was homologue with Klebsiella phage vB_KpP_FBKp27 but only 88% of the genome was covered by this phage. The non-covered parts of the GADU21 genome included genes for tail-fiber-proteins and HNH-endonuclease. GADU22 had 94.8% homology with Escherichia phage vB_Eco_OMNI12 and had genes for immunity proteins. Phylogenetic analysis showed GADU21 and GADU22 were members of Schitoviridae family and Efbeekayvirus genus and Straboviridae family and Tevenvirinae genus, respectively. VIRIDIC analysis classified these phages in new species clusters. Our study demonstrated the possibility to use infected body fluids as phage sources to isolate novel phages. GADU21 is the first reported Klebsiella phage isolated from human body fluid. The absence of virulence and antibiotic resistance genes in their genomes makes the phages a potential therapeutic tool against infections.

Evaluation of antibacterial, antifungal and antibiofilm activities of A. baumannii-derived tannase and gallic acid against uropathogenic microorganisms.

One of the most prevalent infectious diseases and a key driver of antibiotic prescriptions in pediatrics is urinary tract infection (UTI). Due to the emergence of more resistant uropathogenic bacterial and fungal strains, current treatments are no longer effective, necessitating the urgent development of novel antibacterial and antifungal drugs. In this study, the antifungal, antibacterial, and anti-biofilm capabilities of compounds, such as tannase (TN) and gallic acid (GA), which were produced from a novel natural source, Acinetobacter baumannii (AB11) bacteria, were assessed for the inactivation of uropathogenic microorganisms (UMs). Ammonium sulphate precipitation, ion exchange, high-performance liquid chromatography, and gel filtration were used to purify TN and GA that were isolated from A. baumannii. A 43.08 % pure TN with 1221.2 U/mg specific activity and 10.51 mg/mL GA was obtained. The antibacterial, antifungal and anti-biofilm activities of TN and GA were evaluated against UMs and compared to those of commercially available antibiotics including sulfamethoxazole (SXT), levofloxacin (LEV), ciprofloxacin (CIP), amikacin (Ak), and nitrofurantoin (F). The results showed that TN and GA were superior to commercial antibiotics in their ability to inactivate UMs and considerably reduced biofilms formation. Additionally, the GA emerges as the top substitute for currently available medications, demonstrating superior antibacterial and antibiofilm properties against all UMs evaluated in this study. The results of this investigation showed that A. baumannii-derived TN and GA could be utilized as an alternative medication to treat UTIs.

Comparison of three doses of amikacin on alternate days with a daily dose of meropenem during the same period for the treatment of urinary tract infection with E. coli: a double-blind clinical trial.

OBJECTIVES: Urinary tract infections (UTIs) are very common infections in humans, and Escherichia coli (E. coli) is the commonest pathogen leading to UTIs. The generation of beta-lactamase enzymes in this bacterium results in its resistance against many antibiotics. This study compares three doses of amikacin on alternate days with a daily dose of meropenem in the same period for the treatment of UTIs with E. coli in a double-blind clinical trial. METHODS: The current double-blind clinical trial compares three doses of amikacin on alternate days with a daily dose of meropenem in the same period for the treatment of UTIs with E. coli. The patients were assigned to two groups: Intervention (receiving a single dose of amikacin once a day at 48-h intervals for a week, three doses) and control (receiving meropenem for 1/TDS for a week). RESULTS: The E. coli infection frequency was 61 (21 cases of non-ESBL and 40 cases of ESBL-positive infections) and the frequency of the other infections was 52 (46%). In the patients with ESBL E. coli infection, ciprofloxacin (21; 70%) showed the highest antibiotic resistance, and nitrofurantoin (33; 91.7%) showed the highest sensitivity. The baseline variables between the control and intervention groups indicated no significant difference (p > 0.05). The frequency of signs and symptoms showed no significant difference between the amikacin and meropenem groups in the first 24 h and the first week. In the second week of follow-up, no clinical signs or symptoms were observed in the two groups. CONCLUSION: The results of this study showed that treatment with amikacin, 1 g q48h, for one week (three doses) has the same result as meropenem, 1 g q8h, for one week (21 doses). The results are the same for the treatment of UTIs with ESBL positive and ESBL negative. Amikacin can be used once every 48 h to treat UTIs, is less expensive and can be administered on an outpatient basis. TRIAL REGISTRATION: This study was registered in the Iranian Registry of Clinical Trials (IRCT) with ID number: IRCT20170417033483N2 on the date 2018-02-13.

Autovaccine-Based Immunotherapy: A Promising Approach for Male Recurrent Urinary Tract Infections.

BACKGROUND: Recurrent Urinary Tract Infections (UTIs) in men range from 0.9 to 2.4/1000 individuals in younger men to 7.7/1000 in those over 85, significantly impacting their quality of life. Preventive strategies include autovaccines, but limited evidence exists for males. METHODS: A prospective monocentric, open-label observational study was conducted from August 2018 to August 2021, with follow-up until August 2023 including patients with recurrent UTIs treated with immunotherapy. We evaluated the incidence rate of UTIs per year, the incidence rate of episodes after two or three rounds of the autovaccine, and quality of life measured with the IPSS-QoL questionnaire. RESULTS: A total of 49 patients fulfilled inclusion criteria. The mean age was 72 years (±15), and the median 61. The evolution of UTIs number of episodes after the autovaccine rounds: -37.74% for the first round from 5.3 to 3.3; -33.33% for the second round from 3.3 to 2.2; -45.45% for the third round from 2.2 to 1.2. The mean IPSS score improved from 10.69 to 7.27 after the treatment (32%). The mean QoL subscore enhancement was from 4.22 to 1.92 (54%). With a mean follow-up of 3 years, only nine patients required retreatment. CONCLUSION: Autovaccine treatment significantly reduced the number of UTI episodes, with a cumulative effect observed after multiple rounds of treatment, demonstrating an enhancement in QoL and with sustained effectiveness and a low need for retreatment.

Update on Urinary Tract Infection Antibiotic Resistance-A Retrospective Study in Females in Conjunction with Clinical Data.

Urinary tract infections (UTIs) represent a frequent pathology among the female population that has become more and more difficult to treat in the past decade, considering the increase in antibiotic resistance-a serious global public health problem. A cross-sectional retrospective study was conducted for six months to report an update regarding the rates of resistance and susceptibility of uropathogens necessary for optimal treatment. A total of 5487 patients were screened, of which 524 (9.54%) were female patients who met the criteria for inclusion in the study. Escherichia coli was the most common pathogen, representing 290 cases (55.34%), followed by Enterococcus spp. 82 (15.64%). Escherichia coli presented the highest resistance to amoxicillin-clavulanic acid (R = 33.1%), followed by trimethoprim-sulfamethoxazole (R = 32.41%) and levofloxacin (R = 32.06%). The highest sensitivity rates were observed for fosfomycin (S = 96.55%), followed by imipenem (S = 93.1%). Enterococcus spp. showed the highest resistance to levofloxacin (R = 50.0%), followed by penicillin (R = 39.02%). The highest sensitivity was observed for fosfomycin (S = 90.24%), linezolid (S = 89.02%), and nitrofurantoin (S = 86.58%). The second most frequent Gram-negative uropathogen was represented by Klebsiella spp., which had the highest resistance to amoxicillin-clavulanic acid (R = 35.89%), followed by levofloxacin (R = 25.64) and trimethoprim-suflamethoxazole (R = 24.35%). The most frequently associated pathology was an episode of UTI in the previous year, followed by diabetes and chronic kidney disease. Antibiotic resistance is a serious problem for all clinicians who treat UTIs. An up-to-date knowledge of antibiotic resistance rates is a major necessity to stop its evolution. Overall, the highest resistance rates were observed for aminopenicillins, fluoroquinolones, and trimethoprim-sulfamethoxazole. The best susceptibility rates were observed for fosfomycin, nitrofurantoin, and carbapenems. Our report aims to guide clinicians whenever they are forced to prescribe antibiotics empirically.

Characterization and synergy studies of Caudoviricete Escherichia phage FS2B infecting multi-drug resistant uropathogenic Escherichia coli isolates.

Escherichia coli is one of the most common causes of urinary tract infections. However, a recent upsurge in antibiotic resistance among uropathogenic E. coli (UPEC) strains has provided an impetus to explore alternative antibacterial compounds to encounter this major issue. In this study, a lytic phage against multi-drug-resistant (MDR) UPEC strains was isolated and characterized. The isolated Escherichia phage FS2B of class Caudoviricetes exhibited high lytic activity, high burst size, and a small adsorption and latent time. The phage also exhibited a broad host range and inactivated 69.8% of the collected clinical, and 64.8% of the identified MDR UPEC strains. Further, whole genome sequencing revealed that the phage was 77,407 bp long, having a dsDNA with 124 coding regions. Annotation studies confirmed that the phage carried all the genes associated with lytic life cycle and all lysogeny related genes were absent in the genome. Further, synergism studies of the phage FS2B with antibiotics demonstrated a positive synergistic association among them. The present study therefore concluded that the phage FS2B possesses an immense potential to serve as a novel candidate for treatment of MDR UPEC strains.

Evaluation of MV140 in preventing recurrent urinary tract infections: a multicentre double-blind randomized controlled trial protocol.

OBJECTIVES: To determine, firstly, whether MV140 reduces rates of recurrent urinary tract infections (rUTIs) in patients older than 65 years, measured as the number of urinary tract infections (UTIs) detected over 12 months following the completion of a 3-month treatment course and, additionally, to assess the number of re-admissions to the emergency department, the rate of antibiotic use for UTIs, the safety profile of MV140, and quality of life. MATERIALS AND METHODS: This is a multicentre, double-blind, randomized controlled trial with two arms. Patients will be randomized and allocated to receive either a 3-month course of MV140 or placebo (two sublingual sprays daily). Participants will have 3-monthly consultations with the investigator for 12 months to assess differences in rates of rUTIs between the two groups. Study candidates will be identified and recruited from inpatient and outpatient clinics across Sydney via referral to the investigation team. After obtaining consent, participants will undergo initial study consultations including urine microscopy and culture, uroflowmetry, and bladder scan to assess postvoid residual urine volume. Participants will be randomized and provided with a unique trial number. Electronic medical records will be reviewed to collect relevant information. Participants will be provided with a study diary to record relevant data. RESULTS: Follow-up consultations will be conducted every 3 months for a 12-month duration, during which the study diary will be reviewed. These follow-up consultations will primarily occur via telephone review, however, there will be flexibility for in-person reviews for participants who find telephone consultation prohibitively difficult. CONCLUSION: This is a multicentre, double-blinded, randomised control trial, the first in Australia to assess the safety and efficacy of MV140 Uromune vaccine in prevention of recurrent UTIs. Results have been promissing in the global literatures.

Designing of fragment based inhibitors with improved activity against E. coli AmpC ß-lactamase compared to the conventional antibiotics.

One of the most common primary resistance mechanism of multi-drug resistant (MDR) Gram negative pathogenic bacteria to combat ß-lactam antibiotics, such as penicillins, cephalosporins and carbapenems is the generation of ß- lactamases. The uropathogenic E. coli is mostly getting multi-drug resistance due to the synthesis of AmpC ß-lactamases and therefore new antibiotics and inhibitors are needed to treat the evolving infections. The current study was designed for targetting AmpC ß-lactamase of E. coli using molecular docking based virtual screening, linking fragments for designing novel compounds and binding mode analysis using molecular dynamic simulation with target protein. The FCH group all-purpose fragment library consisting of 9388 fragments has been screened against AmpC ß-lactamase protein of E. coli and the antibiotics and anti-infectives used in treatment of Urinary tract Infections (UTIs) were also screened with AmpC ß-lactamase protein. Among the 9388 fragments, 339 fragment candidates were selected and linked with cefepime antibiotic having maximum binding affinity for AmpC target protein. Computational analysis of interactions as well as molecular dynamics (MD) simulations were also conducted for identifying the most promising ligand-pocket complexes from docking investigations to comprehend their thermodynamic properties and verify the docking outcomes as well. Overall, the linked complexes (LCs) showed good binding interactions with AmpC ß-lactamase. Interestingly, our fragment-based LCs remained relatively stable in comparison with cefepime antibiotic. Moreover, S12 fragment linked complex remained the most stable during 50 ns with remarkable number of interactions indicating it as promising candidate in novel lead discovery against MDR E. coli infections.