Urinary Tract Infection Samples Research Articles

Analyzing Antibiotic Resistance: A Comparative Study of E. coli Isolates from Wastewater and Clinical UTI Samples at a University Community in Grenada, West Indies

Abstract Introduction/Objective The rise of antibiotic resistance poses a significant global health threat, needing a robust monitoring system to track its emergence and spread. However, most developing countries lack the infrastructure to monitor such resistance effectively. Wastewater-based epidemiology (WBE) has been proven effective in surveilling various outbreaks and offers a promising method for monitoring antibiotic resistance at the community level. This study aims to assess and compare the antibiotic resistance of Escherichia coli isolated from raw wastewater with those isolated from clinical urinary tract infection (UTI) samples from the same community. Methods/Case Report Conducted from January 2022 to October 2023, this study analyzed antibiogram data from eighty-six Escherichia coli isolates sourced from wastewater and thirty-four isolates derived from clinical urinary tract infections (UTIs). The clinical samples were obtained and processed at the Clinical Microbiology Laboratory of a university campus. The isolates were evaluated against the eleven most prescribed antibiotics for UTIs in the University Clinic including: Amoxicillin/Clavulanic Acid, Ampicillin, Cefazolin, Cefoxitin, Ceftriaxone, Cefuroxime, Ciprofloxacin, Gentamicin, Nalidixic Acid, Nitrofurantoin, and Sulfamethoxazole-trimethoprim. Results (if a Case Study enter NA) Explaining the observed trends in both wastewater and clinical samples from the perspective of AWaRE-WHO classification (Access, Watch, Reserve), we see similar trends of resistance in both data sets. A) The antibiotics used in the Access category (Empiric first or second choice for treatment of most common infections) demonstrates similar resistance patterns for both wastewater and clinical isolates except for Nitrofurantoin. B) The antibiotics used in the Watch category (Antibiotics that have higher toxicity issues and higher potential to negatively impact AMR) resistance pattern shows negligible risk in the university community evaluated. Conclusion It is recommended that the university clinic prescribe Nitrofurantoin as the last antibiotic from the Access list to the university community to treat UTI patients. Furthermore, WBE is a useful tool to determine patterns of resistance under the studied settings.

Molecular detection of Pseudomonas aeruginosa isolated from children with urinary tract infections

The purpose of this research was to identify Pseudomonas aeruginosa from pediatric UTI samples using molecular methods. Bacteria were isolated and identified using a primary medium. Children from all around Baghdad city who were hospitalized at a private clinic between 2019 to 2022 and showed symptoms of a urinary tract infection (UTI) such as fever and vomiting, had their urine sampled. There were 132 samples of children's urine collected from kids who had UTIs. The VITEK-2 system was completed on pure bacterial isolates. DNA of these bacteria was done and two primers were used in this study for the detection of P. aeruginosa. The present study showed that the rate of infection by P. aeruginosa was 7.6%, the total Gram-positive bacteria were 22.7% and the total Gram-negative bacteria were 77.3%. It was shown that females had a greater rate of UTI (55.3% vs. 44.7%) than males. In conclusion, because of its prevalence and severity in community-acquired UTIs in children and its unique risk factors and outcomes, P. aeruginosa should be suspected in any child who has recently received or is now receiving antibiotic treatment.

Multiplex antibiotic susceptibility testing of urinary tract infections using an electrochemical lab-on-a-chip

Urinary tract infections (UTIs) represent the most prevalent type of outpatient infection, with significant adverse health and economic burdens. Current culture-based antibiotic susceptibility testing can take up to 72 h resulting in ineffective prescription of broad-spectrum antibiotics, poor clinical outcomes and development of further antibiotic resistance. We report an electrochemical lab-on-a-chip (LOC) for testing samples against seven clinically-relevant antibiotics. The LOC contained eight chambers, each housing an antibiotic-loaded hydrogel (cephalexin, ceftriaxone, colistin, gentamicin, piperacillin, trimethoprim, vancomycin) or antibiotic-free control, alongside a resazurin bulk-modified screen-printed electrode for electrochemical detection of metabolically active bacteria using differential pulse voltammetry. Antibiotic susceptibility in simulated UTI samples or donated human urine with either Escherichia coli or Klebsiella pneumoniae could be established within 85 min. Incorporating electrochemical detection onto a LOC provides an inexpensive, simple method for the sensitive determination of antibiotic susceptibility that is significantly faster than using a culture-based approach.

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.

Specific GAG ratios in the diagnosis of mucopolysaccharidoses.

Mucopolysaccharidoses (MPS) screening is tedious and still performed by analysis of total glycosaminoglycans (GAG) using 1,9-dimethylmethylene blue (DMB) photometric assay, although false positive and negative tests have been reported. Analysis of differentiated GAGs have been pursued classically by gel electrophoresis or more recently by quantitative LC-MS assays. Secondary elevations of GAGs have been reported in urinary tract infections (UTI). In this manuscript, we describe the diagnostic accuracy of urinary GAG measurements by LC-MS for MPS typing in 68 untreated MPS and mucolipidosis (ML) patients, 183 controls and 153 UTI samples. We report age-dependent reference values and cut-offs for chondroitin sulfate (CS), dermatan sulfate (DS), heparan sulfate (HS) and keratan sulfate (KS) and specific GAG ratios. The use of HS/DS ratio in combination to GAG concentrations normalized to creatinine improves the diagnostic accuracy in MPS type I, II, VI and VII. In total 15 samples classified to the wrong MPS type could be correctly assigned using HS/DS ratio. Increased KS/HS ratio in addition to increased KS improves discrimination of MPS type IV by excluding false positives. Some samples of UTI patients showed elevation of specific GAGs, mainly CS, KS and KS/HS ratio and could be misclassified as MPS type IV. Finally, DMB photometric assay performed in MPS and ML samples reveal four false negative tests (sensitivity of 94%). In conclusion, specific GAG ratios in complement to quantitative GAG values obtained by LC-MS enhance discrimination of MPS types. Exclusion of patients with UTI improve diagnostic accuracy in MPS IV but not in other types.

Targeting Superbugs: Efficacy of Bacteriophage Therapy against Antibiotic-Resistant Pseudomonas Aeruginosa in Urinary Tract Infections

The rise of antibiotic-resistant bacterial infections poses a significant challenge to global health, particularly in the treatment of urinary tract infections (UTIs). This study evaluates the potential of bacteriophage therapy as an alternative treatment against antibiotic-resistant Pseudomonas Aeruginosa isolated from UTI samples. We isolated 12 strains of P. aeruginosa from 50 UTI samples and conducted extensive antibiotic susceptibility testing, revealing significant resistance to multiple conventional antibiotics such as Tetracycline, Septran, Ceftazidime, and Cefepime, while showing susceptibility to Imipenem and Meropenem. Concurrently, we isolated and characterized bacteriophages from sewage samples that demonstrated specific lytic activity against the antibiotic-resistant strains. Our findings suggest that bacteriophage therapy provides a high specificity and efficacy in targeting antibiotic-resistant P. aeruginosa, offering a promising alternative to traditional antibiotic therapies. This study underscores the potential of phages in clinical applications, advocating for further clinical trials and the development of a regulatory framework to integrate phage therapy into mainstream medical practice for combating antibioticresistant infections.

Combining droplet microfluidics and magnetoresistive sensors for the rapid and quantitative detection of Klebsiella pneumoniae in urinary tract infections

Urinary tract infections (UTIs) are one of the most common bacterial infections and are considered a public health problem. This has been exacerbated due to the increasing prevalence of multidrug-resistant bacterial strains which poses a significant concern. Additionally, current diagnostic methods are time-consuming and laborious, hindering appropriate treatment and infection control. Herein a novel microfluidic system for diagnosis of UTIs caused by Klebsiella pneumoniae is presented, relying on the unprecedented combination of magnetoresistive (MR) sensors with droplet microfluidics. Initially, K. pneumoniae suspensions were encapsulated in droplets with magnetic nanoparticles (MNPs) functionalized with a bacteriophage receptor binding protein (RBP) specific for K. pneumoniae. The magnetically labeled cells inside droplets were further detected through an MR readout system. This strategy enabled the refined control over the number of MNPs and bacteria inside each droplet which allowed the clear discrimination between the output signals from samples of K. pneumoniae and non-target Escherichia coli. In addition, a direct correlation of the number of events from the MR sensors with different concentrations of K. pneumoniae was observed, providing a dynamic detection range of this pathogen from 104 to 108 CFU/mL. The developed device was further validated for the detection of K. pneumoniae in urine clinical samples (UCSs) from patients with UTIs. These samples were directly encapsulated with RBP-functionalized MNPs, with no need for sample pre-treatment. This allowed the quantitative detection of K. pneumoniae in UTIs samples with high specificity (100%) and sensitivity (87.5%) in less than 1 h.

Rapid Screening of Urinary Tract Infection Using Microfluidic Inertial-Impedance Cytometry.

Urinary tract infection (UTI) diagnosis based on urine culture for bacteriuria analysis is time-consuming and often leads to wastage of hospital resources due to false-positive UTI cases. Direct cellular phenotyping (e.g., RBCs, neutrophils, epithelial cells) of urine samples remains a technical challenge as low cell concentrations, and urine characteristics (conductivities, pH, microbes) can affect the accuracy of cell measurements. In this work, we report a microfluidic inertial-impedance cytometry technique for label-free rapid (<5 min) neutrophil sorting and impedance profiling from urine directly. Based on size-based inertial focusing effects, neutrophils are isolated, concentrated, and resuspended in saline (buffer exchange) to improve consistency in impedance-based single-cell analysis. We first observed that both urine pH and the presence of bacteria can affect neutrophil high-frequency impedance measurements possibly due to changes in nucleus morphology as neutrophils undergo NETosis and phagocytosis, respectively. As a proof-of-concept for clinical testing, we report for the first time, rapid UTI testing based on multiparametric impedance profiling of putative neutrophils (electrical size, membrane properties, and distribution) in urine samples from non-UTI (n = 20) and UTI patients (n = 20). A significant increase in cell count was observed in UTI samples, and biophysical parameters were used to develop a UTI classifier with an area under the receiver operating characteristic curve of 0.84. Overall, the developed platform facilitates rapid culture-free urine screening which can be further developed to assess disease severity in UTI and other urologic diseases based on neutrophil electrical signatures.

Specific Instantaneous Detection of Klebsiella pneumoniae for UTI Diagnosis with a Plasmonic Gold Nanoparticle Conjugated Aptasensor.

Urinary tract infection (UTI), which can be caused by various pathogens, if not detected at an early stage can be fatal. It is essential to identify the specific pathogen responsible for UTI for appropriate treatment. This study describes a generic approach to the fabrication of a prototype for the noninvasive detection of a specific pathogen using a tailor-made plasmonic aptamer-gold nanoparticle (AuNP) assay. The assay is advantageous because the adsorbed specific aptamers passivate the nanoparticle surfaces and reduce and/or eliminate false-positive responses to nontarget analytes. Based on the localized surface plasmon resonance (LSPR) phenomena of AuNP, a point-of-care aptasensor was designed that shows specific changes in the absorbance in the visible spectra in the presence of a target pathogen for robust and fast screening of UTI samples. In this study, we demonstrate the specific detection of Klebsiella pneumoniae bacteria with LoD as low as 3.4 × 103 CFU/mL.

Epidemiology of Bacterial Urinary Tract Infection and Antibiotics Sensitivity among Pregnant Women at Taiz City, Yemen

Background: Bacterial Urinary Tract Infections (UTIs) is an infection caused by the presence and growth of bacteria anywhere in the urinary tract. Objective: The present study was done to evaluate epidemiology of UTI (prevalence, risk factors, bacterial isolates and antibiotic sensitivity) among pregnant women in Taiz, City, Yemen. Methods: A cross-sectional study design was used for this research from March 2022 to July 2022, including 200 outpatient cases of pregnant women, obtained from hospitals in Taiz-city, Yemen. Urine specimens of pregnant women using midstream urine. Bacteria were identified by standard microbiological methods. Results: Escherichia coli was the most prevalent isolate, it was recovered 36 (42.86%) of total bacterial UTIs samples, followed by Staphylococcus aureus as the second most common representing in 24 (28.57%), and Klebsiella sp was appeared in 15 (17.86%), the Proteus sp was representing in 5 (5.95%), and Staphylococcus saprophyticus appeared in 4 (4.76%). The remaining urine sample isolates were non-bacterial growth (negative cases), representing 116 (58%). Bacterial UTIs were highly present in the age group between 25-34 years, secondary education patients, housewife patients, third pregnancy or above, and during the third trimester. Escherichia coli isolates were (100%) sensitive to Tobramycin, Ceftazidime, Ceftriaxone and Aztreonam. Klebsiella sp isolates were (100%) sensitive to Tobramycin only. Proteus sp isolates were (100%) sensitive to Tobramycin, Ceftazidime, and Ceftriaxone. Staphylococcus saprophyticus isolates were (100%) sensitive to all antibacterial drugs except Methiciliin. Staphylococcus aureus isolates were (100%) sensitive to Tobramycin and Aztreon. Conclusions: Escherichia coli washwhfhj the most prevalent isolate from urine of pregnant women. The most important risk factors are third pregnancy, third trimester, and symptomatic bacteruria. Most of the tested antibiotics showed sensitivity to the bacterial isolates. Urine culture for screening and diagnosis purpose for all pregnant is recommended.

MC-PRPA-HLFIA Cascade Detection System for Point-of-Care Testing Pan-Drug-Resistant Genes in Urinary Tract Infection Samples.

Recently, urinary tract infection (UTI) triggered by bacteria carrying pan-drug-resistant genes, including carbapenem resistance gene blaNDM and blaKPC, colistin resistance gene mcr-1, and tet(X) for tigecycline resistance, have been reported, posing a serious challenge to the treatment of clinical UTI. Therefore, point-of-care (POC) detection of these genes in UTI samples without the need for pre-culturing is urgently needed. Based on PEG 200-enhanced recombinase polymerase amplification (RPA) and a refined Chelex-100 lysis method with HRP-catalyzed lateral flow immunoassay (LFIA), we developed an MCL-PRPA-HLFIA cascade assay system for detecting these genes in UTI samples. The refined Chelex-100 lysis method extracts target DNA from UTI samples in 20 min without high-speed centrifugation or pre-incubation of urine samples. Following optimization, the cascade detection system achieved an LOD of 102 CFU/mL with satisfactory specificity and could detect these genes in both simulated and actual UTI samples. It takes less than an hour to complete the process without the use of high-speed centrifuges or other specialized equipment, such as PCR amplifiers. The MCL-PRPA-HLFIA cascade assay system provides new ideas for the construction of rapid detection methods for pan-drug-resistant genes in clinical UTI samples and provides the necessary medication guidance for UTI treatment.

Evaluating the association between expanded coverage of direct-to-consumer telemedicine and downstream utilization and quality of care for urinary tract infections and sinusitis.

To compare direct-to-consumer (DTC) telemedicine and in-person visits in rates of testing, follow-up health care use, and quality for urinary tract infections (UTIs) and sinusitis. The Minnesota All Payer Claims Data provided 2008-2015 administrative claims data. Using a difference-in-differences approach, we compared episodes of care for UTIs and sinusitis among enrollees of health plans introducing coverage for DTC telemedicine relative to those without DTC telemedicine coverage. Primary outcomes included number of laboratory tests, antibiotics filled, office and outpatient visits, emergency department (ED) visits, and standardized spending, based on standardized prices of health services. The study sample included non-elderly enrollees of commercial health insurance plans. We constructed 30-day episodes of care initiated by a DTC telemedicine or in-person visit. The UTI and sinusitis samples were comprised of 215,134 and 624,630 episodes of care, respectively. Following the introduction of coverage for DTC telemedicine, 15.7% of UTI episodes and 8.9% of sinusitis episodes were initiated with DTC telemedicine. Compared to episodes without coverage for DTC telemedicine, UTI episodes with coverage had 0.25 fewer lab tests (95% CI: -0.33, -0.18; p < 0.001), lower standardized spending for the first UTI visit (-$11.18 [95% CI: -$21.62, -$0.75]; p < 0.05), and no change in office and outpatient visits, ED visits, antibiotics filled, or standardized medical spending. Sinusitis episodes with coverage for DTC telemedicine had fewer antibiotics filled (-0.08 [95% CI: -0.14, -0.01]; p < 0.05) and a very small increase in ED visits (0.001 [95% CI: 0.001, 0.010]; p < 0.05), but no change in lab tests, office and outpatient visits, or standardized medical spending. Among commercially insured patients, coverage of DTC telemedicine was associated with reductions in antibiotics for sinusitis and laboratory tests for UTI without changes in downstream total office and outpatient visits or changes in ED visits.

Antibiotic Resistance Profiling of Pathogenic Staphylococcus Species from Urinary Tract Infection Patients in Benin.

Staphylococci can cause urinary tract infections (UTIs). These UTIs are among the significant causes of antibiotic resistance and the spread of antibiotic-resistant diseases. The current study is aimed at establishing a resistance profile and determining the pathogenicity of Staphylococcus strains isolated from UTI samples collected in Benin. For this purpose, urine samples (one hundred and seventy) that were collected from clinics and hospitals showed UTI in patients admitted/visited in Benin. The biochemical assay method was used to identify Staphylococcus spp., and the disk diffusion method tested the antimicrobial susceptibility. The biofilm formation ability of the isolates of Staphylococcus spp. was investigated by the colorimetric method. The presence of mecA, edinB, edinC, cna, bbp, and ebp genes was examined by multiplex polymerase chain reaction (PCR). The results showed that Staphylococcus species were identified in 15.29% of all infected individuals and that 58% of these strains formed biofilms. Most Staphylococcus strains (80.76%) were isolated in female samples, and the age group below 30 years appeared to be the most affected, with a rate of 50%. All Staphylococcus strains isolated were 100% resistant to penicillin and oxacillin. The lowest resistance rates were seen with ciprofloxacin (30.8%), gentamicin, and amikacin (26.90%). Amikacin was the best antibiotic against Staphylococcus strains isolated from UTIs. The isolates carried mecA (42.31%), bbp (19.23%), and ebp (26.92%) genes in varying proportions. This study provides new information on the risks posed to the population by the overuse of antibiotics. In addition, it will play an essential role in restoring people's public health and controlling the spread of antibiotic resistance in urinary tract infections in Benin.

Evaluation and Quantification of Resistant Gene in Antibiotic Resistant ESBL Producing E. coli in UTI Samples with Diabetes Mellitus: A Molecular Study

In diabetic patients, urinary tract infections (UTIs) are more common and severe often caused by antibiotic resistance and carry worse consequences, due to high blood sugar level and frequent urination. High blood sugar provides favorable environment for the growth of pathogens and some other impairment in immune system, poor metabolic control and incomplete emptying bladder to enhance the risk of UTIs in the diabetic patients. Most of the UTIs are caused by multidrug resistant, Extended Spectrum β lactamase (ESBL) producing bacteria mostly, Escherichiacoli (E. coli). The prevalence of UTIs caused by ESBL producing Multidrug Resistant (MDR) bacteria is a major global public health concern. The aim of this study was to identify the pattern of multidrug resistance and quantification of resistance genes amongst the E. coli isolates which were obtained from UTI patients with diabetes. ESBL producing stains were found 36% by using amoxicillin/clavulanic acid as a β lactamase inhibitor. Out of these stains, following antibiotics resistance rates were achieved with ampicillin, amoxicillin, amikacin, cefotaxime, ceftazidime, chloramphenicol, ciprofloxacin, gentamicin, nitrofurantoin, norfloxacin, tetracycline and amoxicillin/clavulanic acid as 100%, and 0%, respectively. The ESBL encoding genes were characterized for CTX-M, TEM and SHV genes. CTX-M gene was found to have resistance in the E. coli isolates, some others genes as TEM and SHV respectively. These characterized genes were analyzed through PCR which is a rapid, sensitive and easy to interpret detection method. Among these isolates, CTX-M was significantly higher 85.4% in the ESBL positive strains (p &lt; 0.05). TEM and SHV were found 12.8% and 6.5%, respectively. Early and proper diagnosis is necessary for the management of UTIs in diabetic patients in order to have an adequate treatment.

Whole genome global insight of antibiotic resistance gene repertoire and virulome of high - risk multidrug-resistant Uropathogenic Escherichiacoli

Elucidation of genetic determinants via whole genome sequence (WGS) analyses can help understand the high risk multidrug-resistant (MDR) Uropathogenic Escherichia coli (UPEC) associated with urinary tract infections (UTI) and its evasion strategies from treatment. We investigated the WGS of 30 UPEC strains from UTI samples across the world (2016–2019) and found 25 UPEC strains carrying 2–23 antibiotic resistance genes (ARGs) scattered across 1–3 plasmids per strain. Different ARGs (blaTEM, blaCTXM, blaNDM, blaOXA, blaCMY) encoding extended-spectrum beta-lactamases (TEM, CTXM, CMY) and carbapenemases (NDM, OXA) were found in 24/30, ARGs encoding aminoglycoside modifying enzymes (AAC, APH, AAD) variants in 23/30, trimethoprim ARGs (dfrA17, dfrA12, dfrA5, dfrB4 variants) encoding dihydrofolate reductase in 19/30 and sulfonamide ARGs (sul1, sul2, sul3) encoding dihydropteroate synthase and macrolide ARGs (mph1) encoding macrolide 2’ phosphotransferase in 15/30 UPEC strains. Collectively the ARGs were distributed in different combinations in 40 plasmids across UPEC strains with 20 plasmids displaying co-occurrence of multiple ARGs conferring resistance to beta lactam, aminoglycoside, sulfonamide, trimethoprim and macrolide antibiotics. These resistance plasmids belonged to seven incompatibility groups (IncF, IncI, IncC, IncH, IncN, IncB and Col), with IncFI and IncFII being the predominant resistance plasmids. Additionally, we observed co-occurrence of specific mutation pattern in quinolone resistance determining region (QRDR) viz., DNA gyrase (gyrA: S83L, D87N), and topoisomerase IV (parC: S80I, E84V; parE: I529L) in 18/30 strains. The strains also harbored diverse virulence genes, such as fimH, gad, iss, iha, ireA, iroN, cnf1 and san. Multilocus sequence typing (MLST) reconfirmed ST131(n = 10) as the predominant global high-risk clonal strain causing UTI. In summary, our findings contribute to better understand the plasmid mediated ARGs and its encoded enzymes that may contribute in antibiotic inactivation/modification or alteration in the antibiotic target site in high risk MDR hypervirulent UPEC strains causing UTI. The study reinforces the need to characterize and design appropriate inhibitors to counterattack different enzymes and devise strategies to curtail resistance plasmid.

Frequency of hlyA, hlyB, hlyC and hlyD genes in uropathogenic Escherichia coli isolated from UTI patients in Shiraz.

Background and objectives: One of the most important causes of urinary tract infections (UTI) is Escherichia coli. The infection is mainly due to the uropathogenic strain (UPEC), which has key virulence factors, including hemolysis. In this study, we evaluated the frequency of hlyA, hlyB, hlyC and hlyD genes in UPEC strains isolated from clinical samples from Shiraz city, Iran.Materials and methods: 130 urine samples with suspected UTI were collected from Shiraz medical centers and cultured on blood agar and EMB media. Colonies were then characterized by biochemical methods. The genomes were extracted and the presence of hemolysis genes was detected by polymerase chain reaction (PCR) using hly gene specific primers and 16S rRNA. Drug resistance was assessed by using 10 antibiotic disks in the disk diffusion method, according to CLSI criteria.Results: Out of the 130 collected UTI samples, 100 were identified as UPECs. Within isolates, the hlyD gene had the highest frequency – 95% – and hlyC had the lowest, with 23%. The frequencies of hlyA and hlyB genes were calculated as 50% and 43%, respectively. The rates of antibiotic resistance to Azithromycin, Ampicillin, Cefotaxime, Nalidixic Acid, Tetracycline, Trimethoprim-Sulfamethoxazole, Cefepime, Aztreonam, Gentamicin, and Nitrofurantoin were 95%, 86%, 68%, 66%, 65%, 64%, 51%, 46%, 44%, 14%, respectively. 98% of these isolates belonged to the MDR group.Conclusion: This study shows diversity of hemolysis virulence factor in UPECs and unique UPEC drug resistance that would indicate a high antibiotic use in the general population.

Asymptomatic Bacteriuria (ABU) in Elderly: Prevalence, Virulence, Phylogeny, Antibiotic Resistance and Complement C3 in Urine.

Background: The incidence of asymptomatic bacteriuria (ABU) increases with age and is most common for persons 80 years of age and above and in elderly living in nursing homes. The distinction between ABU and urinary tract infection (UTI) is often difficult, especially in individuals, who are unable to communicate their symptoms, and there is a lack of objective methods to distinguish between the two entities. This can lead to overuse of antibiotics, which results in the selection and dissemination of antibiotic resistant isolates. Materials and methods: From voided midstream urine samples of 211 participants ≥60 years old from nursing homes, an activity center and a general practitioners clinic, we collected 19 ABU, 16 UTI and 22 control urine samples and compared them with respect to levels of complement component C3 in urine as determined by an ELISA assay relative to creatinine levels in the same urine samples, as measured by a creatinine assay. Further, we studied all Escherichia coli isolates for selected virulence genes by multiplex PCR, and by whole-genome sequencing (WGS) for genotypes and phylogenetic clustering. Antibiotic susceptibility was determined by microtiter broth dilution. Results: We identified a prevalence of ABU of 18.9% in nursing home residents, whereas ABU was only found in 4% of elderly living in the community (p < 0.001). E. coli from ABU patients were significantly more antibiotic resistant than E. coli from UTIs (p = 0.01). Prevalence of classical virulence genes, detected by multiplex PCR, was similar in E. coli isolates from ABU and UTI patients. Whole-genome sequencing of the E. coli isolates showed no specific clustering of ABU isolates compared to UTI isolates. Three isolates from three different individuals from one of the nursing homes showed signs of transmission. We demonstrated a significantly increased level of C3/creatinine ratio in ABU and UTI samples compared to healthy controls; however, there was no significant difference between the ABU and UTI group with respect to C3 level, or virulence factor genes. Conclusion: ABU was significantly more prevalent in the elderly residing in nursing homes than in the elderly living at home. Antibiotic resistance was more prevalent in E. coli from nursing homes than in UTI isolates, but there was no difference in prevalence of virulence associated genes between the two groups and no phylogenetic clustering, as determined by WGS relative to the two types of E. coli bacteriuria. The similar complement C3 response in ABU and UTI patients may indicate that ABU should be reconsidered as an infection albeit without symptoms.

Transcriptomic analyses and experimental verification reveal potential biomarkers and biological pathways of urinary tract infection

ABSTRACT Urinary tract infection (UTI) is a common infectious disease. Urinary tract pathogenic Escherichia coli (UPEC) is the main cause of UTIs. At present, antibiotics are mainly used for the treatment of UTIs. However, with the increase of drug resistance, the course of the disease is prolonged. Therefore, identifying the receptors and signal pathways of host cells and tissues will further our understanding of the pathogenesis of UTIs and help in the development of new drug treatments. We used two public microarray datasets (GSE43790, GSE124917) in the Gene Expression Omnibus (GEO) database to identify differentially expressed genes (DEGs) between UTI and normal cell samples. A functional analysis based on Gene Ontology (GO) data, a pathway enrichment analysis based on Kyoto Encyclopedia of Genes and Genomes (KEGG) data and a protein-protein interaction analysis identified the main potential biomarkers and verified them in animal tissues. A total of 147 up-regulated genes and 40 down-regulated genes were identified. GO enrichment analysis showed that these functional changes relate to the terms response to lipopolysaccharide, regulation of cytokine production, and regulation of the inflammatory response. KEGG analysis indicated that urinary tract infections likely involve the TNF-αsignaling pathways. The 20 hub genes were selected from the protein-protein interaction network, and the highly significant hub genes were verified by animal experiments. Our findings provide potential targets for exploring new treatments for urinary tract infections. After a comprehensive analysis of the GEO database, these results may facilitate development of new diagnosis and treatment strategies for urinary tract infections.

In Vitro Activity of Imipenem/Relebactam Against Enterobacteriaceae Isolates Obtained from Intra-abdominal, Respiratory Tract, and Urinary Tract Infections in China: Study for Monitoring Antimicrobial Resistance Trends (SMART), 2015-2018.

Considering the increasing incidence of carbapenem-resistant Enterobacteriaceae in China, this study aimed to establish the in vitro effectiveness of imipenem/relebactam (IMI/REL) on clinical Enterobacteriaceae isolates derived from intra-abdominal infections (IAIs), respiratory tract infections (RTIs), and urinary tract infections (UTIs) in China between 2015 and 2018. In total, 8781 Enterobacteriaceae isolates from IAI, RTI, and UTI samples were collected from 22 hospitals across 7 geographic regions of China. Susceptibility to antimicrobial drugs was tested using the Clinical and Laboratory Standards Institute broth microdilution and breakpoints, and IMI/REL activity was assessed using United States Food and Drug Administration guidelines. In 2015-2018, the most frequently identified Enterobacteriaceae species was Escherichia coli (n = 4676 [53.3%]), followed by Klebsiella pneumoniae (n = 2949 [33.6%]) and Enterobacter cloacae (n = 542 [6.2%]). The Enterobacteriaceae isolates showed 95.2% overall susceptibility to IMI/REL, of which the susceptibility rates in isolates from IAI, RTI, and UTI were 95.8%, 91.4%, and 96.6%, respectively. Overall, the susceptibilities of both intensive care unit (ICU) and non-ICU Enterobacteriaceae isolates to colistin were 92.9%, followed by IMI/REL (90.7% [95.9%]) and amikacin (83.3% [92.3%]). In addition, IMI/REL restored 66.3% susceptibility in imipenem-nonsusceptible Enterobacteriaceae. Given their high in vitro susceptibility, Enterobacteriaceae infections in China should be considered for IMI/REL treatment, especially with isolates that are not susceptible to carbapenems.

An Intact Cell Bioluminescence-Based Assay for the Simple and Rapid Diagnosis of Urinary Tract Infection.

Urinary tract infection (UTI) is one of the most common infections, accounting for a substantial portion of outpatient hospital and clinic visits. Standard diagnosis of UTI by culture and sensitivity can take at least 48 h, and improper diagnosis can lead to an increase in antibiotic resistance following therapy. To address these shortcomings, rapid bioluminescence assays were developed and evaluated for the detection of UTI using intact, viable cells of Photobacterium mandapamensis USTCMS 1132 or previously lyophilized cells of Photobacterium leiognathi ATCC 33981™. Two platform technologies—tube bioluminescence extinction technology urine (TuBETUr) and cellphone-based UTI bioluminescence extinction technology (CUBET)—were developed and standardized using artificial urine to detect four commonly isolated UTI pathogens—namely, Escherichia coli, Proteus mirabilis, Staphylococcus aureus, and Candida albicans. Besides detection, these assays could also provide information regarding pathogen concentration/level, helping guide treatment decisions. These technologies were able to detect microbes associated with UTI at less than 105 CFU/mL, which is usually the lower cut-off limit for a positive UTI diagnosis. Among the 29 positive UTI samples yielding 105–106 CFU/mL pathogen concentrations, a total of 29 urine specimens were correctly detected by TuBETUr as UTI-positive based on an 1119 s detection window. Similarly, the rapid CUBET method was able to discriminate UTIs from normal samples with high confidence (p ≤ 0.0001), using single-pot conditions and cell phone-based monitoring. These technologies could potentially address the need for point-of-care UTI detection while reducing the possibility of antibiotic resistance associated with misdiagnosed cases of urinary tract infections, especially in low-resource environments.