Agent Of Urinary Tract Infections Research Articles

Virulence potential of the first Corynebacterium mycetoides strain isolated from human urine: a rare species of Corynebacterium

For many years, the potential pathogenic of non-diphtheriae corynebacteria were underestimated. Nowadays, a growing number of Corynebacterium species are recognized as opportunistic agents of human infections, mainly in hospital settings. In addition, multidrug-resistant Corynebacterium isolates from clinical specimens, have been reported and the role of Corynebacterium spp. in urinary tract infections (UTIs) has been highlighted. Several studies have reported Corynebacterium species as the agent of UTIs especially in patients with risk factors. Thus, the present work aimed to report the first isolation of Corynebacterium mycetoides from human urine and an initial study on its virulence properties. The isolate, initially characterized by phenotypical tests as a multidrug-resistant Corynebacterium sp., was recovered from the urine of a female transplant patient. Mass spectrometry and 16S rRNA and rpoB genes sequencing identified the isolate as C. mycetoides. The isolate was found able to adhere to and survive into epithelial cells (Vero cells), and its pathogenic potential was confirmed when tested against Caenorhabditis elegans nematode. The results obtained suggest that C. mycetoides is a potential pathogen for the urinary tract in humans and for a better understanding of the multifactorial mechanisms of virulence, studies about this species should be continued.

Crude metabolites from endophytic fungi inhabiting Cameroonian Annona muricata inhibit the causative agents of urinary tract infections

Urinary tract infections (UTIs) are common bacterial infections. The global emergence of multidrug-resistant uropathogens in the last decade underlines the need to search for new antibiotics with novel mechanisms of action. In this regard, exploring endophytic fungi inhabiting medicinal plants used locally against urinary tract infections could be a promising strategy for novel drug discovery. This study investigates crude metabolites from endophytic fungi isolated from Annona muricata as potential sources of antibiotic drugs to fight against uropathogens and reduce related oxidative stress. Crude ethyl acetate extracts from 41 different endophytic fungi were screened against three bacterial strains using the broth microdilution method, and fungi producing active crude extracts were identified using ITS1-5.8S rRNA-ITS2 nucleotide sequences. The antibacterial modes of action of the five most active extracts were evaluated using Staphylococcus aureus ATCC 43300 and Klebsiella oxytoca strains. The DPPH and FRAP assays were used to investigate their antioxidant activity, and their cytotoxicity against the Vero cell line was evaluated using the MTT assay. Out of the 41 crude extracts tested, 17 were active with minimum inhibitory concentrations (MICs) ranging from 3.125 μg/mL to 100 μg/mL and were not cytotoxic against Vero cell lines with a cytotoxic concentration 50 (CC50) >100 μg/mL. The more potent extracts (from Fusarium waltergamsii AMtw3, Aspergillus sp. AMtf15, Penicillium citrinum AMf6, Curvularia sp. AMf4, and Talaromyces annesophieae AMsb23) significantly inhibited bacterial catalase activity, lysed bacterial cells, increased outer membrane permeability, and inhibited biofilm formation, and the time-kill kinetic assay revealed concentration-dependent bactericidal activity. All seventeen extracts showed weak ferric iron-reducing power (1.06 to 12.37 μg equivalent NH2OH/g of extract). In comparison, seven extracts exhibited DPPH free radical scavenging activity, with RSA50 ranging from 146.05 to 799.75 μg/mL. The molecular identification of the seventeen active fungi revealed that they belong to six distinct genera, including Aspergillus, Curvularia, Fusarium, Meyerozyma, Penicillium, and Talaromyces. This investigation demonstrated that fungal endophytes from Cameroonian Annona muricata, a medicinal plant used locally to treat bacterial infections, might contain potent antibacterial metabolites with multiple modes of action. The antibacterial-guided fractionation of these active extracts is currently ongoing to purify and characterise potential antibacterial active ingredients.

Glycosuria Alters Uropathogenic Escherichia coli Global Gene Expression and Virulence.

ABSTRACTUropathogenic Escherichia coli (UPEC) is the principal etiology of more than half of urinary tract infections (UTI) in humans with diabetes mellitus. Epidemiological data and studies in mouse model of ascending UTI have elucidated various host factors responsible for increasing the susceptibility of diabetic hosts to UPEC-UTI. In contrast, diabetic urinary microenvironment-mediated alterations in UPEC physiology and its contributions to shaping UPEC-UTI pathogenesis in diabetes have not been examined. To address our central hypothesis that glycosuria directly induces urinary virulence of UPEC, we compared virulence characteristics and gene expression in human UPEC strains UTI89 (cystitis) and CFT073 (pyelonephritis), exposed for 2 h in vitro to urine from either male or female donors that was either plain or supplemented with glucose to mimic glycosuria. Compared to control UPEC exposed to nutrient-rich culture medium, lysogeny broth, glycosuria-exposed UPEC exhibited significant increase in biofilm formation and reduction in the hemagglutination of Guinea pig erythrocytes (a measure of type 1 piliation). In addition, the analysis of UTI89 transcriptome by RNA sequencing revealed that 2-h-long, in vitro exposure to glycosuria also significantly alters expression of virulence and metabolic genes central to urinary virulence of UPEC. Addition of galactose as an alternative carbon source affected biofilm formation and gene expression profile of UPEC to an extent similar to that observed with glucose exposure. In summary, our results provide novel insights into how glycosuria-mediated rapid changes in UPEC fitness may facilitate UTI pathogenesis in the diabetic urinary microenvironment.IMPORTANCE Uropathogenic Escherichia coli (UPEC) is an important causative agent of urinary tract infections in diabetic humans. We examined the effects of in vitro exposure to glycosuria (presence of glucose in urine) on the virulence and gene expression by UPEC. Our results show that glycosuria rapidly (in 2 h) alters UPEC gene expression, induces biofilm formation, and suppresses type 1 piliation. These results offer novel insights into the pathogenesis of UPEC in the urinary tract.

Etiological Agents of Urinary Tract Infection (UTI)

Urinary tract infection is one of the most common infectious diseases and has a high financial burden on society. The occurrence of urinary tract infection acquired by the population is higher in young women, almost half of all women will have at least one UTI episode during their lifetime, and about 1 in 3 women will have at least one UTI episode at the age of 24 years. Urinary tract infection increases with age for both sexes. It is estimated that 10 percent of men and 20 percent of women over 65 years of age have bacterial symptoms. Worldwide, Community-associated UTI (CAUTI) prevalence is 0.7% and the main risk factors are age, sexual activity, history of urinary tract infection, and diabetes. UTI is caused by Gram-negative and Gram-positive bacteria, where more than 95 % of UTI cases are caused by bacteria. Gram-negative organisms, primarily from the Enterobacteriaceae family, are responsible for UTI. It includes Escherichia coli (74.20 %), Enterococcus spp (5.30 %), Staphylococcus saprophyticus (1.40 %) Pseudomonas spp. (3.20 %), Klebsiella pneumoniae (6.20 %), and Proteus mirabilis (2.00 %) are among the bacteria that cause UTIs, with other bacteria accounting for 8.70 % .The aim of this review is to provide a summary and critical evaluation of the published evidence about the etiological agents of urinary tract infection.

Bacterial vaginosis as a risk factor for lower urinary tract infection in women: case report

Conservative therapy of acute (exacerbation of chronic) cystitis in women in combination with bacterial vaginosis is a task that often causes difficulties for a practicing doctor. Escherichia coli is still the main causative agent of urinary tract infection due to the presence of virulence factors that not only affect the affinity of the pathogen to the urothelium, but also hinder the development of an immune response. At the same time, bacterial vaginosis is a factor that often accompanies and supports inflammation of the lower urinary tract in women. Simultaneous occurrence of the above conditions is a frequent clinical situation. A differential approach is needed to prescribe antimicrobials to patients with urinary tract infection and bacterial vaginosis, as well as the use of safe natural medicines (biologically active additives) in their complex therapy.

Study of Antibacterial Chemical Substances and Molecular Investigation among Sulfamethoxazole-trimethoprim (SXT)-Resistant Escherichia coli Isolates.

Escherichia coli (E. coli) remains one of the leading agents of urinary tract infection (UTIs), it has become resistant to many drugs. Current work aimed to evaluate some chemical substances as antibacterial agents and molecular study of virulence factors associated with UTIs. This work involved 133 urine specimens obtained from females' patients suffering from UTIs, Methods of well diffusion and disk diffusion were achieved to assay the effect of some chemical substances and antibiogram profiles toward Sulfamethoxazole-trimethoprim (SXT)-resistant E. coli respectively. Virulence genes were done based on the technique of Polymerase Chain Reaction (PCR). The results recorded 49/133 (36.84%) E. coli among women suffering UTIs, 28/49 (57.14%) were resistant to SXT drug. imipenem, meropenem, and nitrofurantoin were recorded more effectively. Chemicals substances at the concentration 0.3 (g/ml) recorded percentages of inhibition, reaching 9.143±1.442, 15.36±0.5914, and 21.82±0.8699 for NaHCO3, Ch4c, and Viroxide Super™ respectively. PCR demonstrated that 28/28 (100%) of SXT-resistant E. coli isolates were harbored Sul-2, FeoB and PapC genes, while 14/28 (50%), 15/28 (53.57%), 19/28 (67.85%) and 26/28 (92.85%) in U250 (pet), FumC, Sul-1 and IutA genes, respectively. Sul-3 gene was not observed. Observed a high percentage of E. coli that were resistant to SXT drug, and having several virulence genes, poses a real threat, it requires a real pause to create substitutions to limit the spreading of this threat.

Genotypic Detection of qnrA and qnrC Genes in Citrobacter koseri Isolated from Patients with Urinary Tract Infection.

The urinary tract infection (UTI) is a prevalent infection that affects people of all ages. Bacterial agents are the most common causes of UTIs. Klebsiella pneumoniae, Proteus mirabilis, Pseudomonas aeruginosa, and other staphylococcal species, Citrobacter freundii and Citrobacter koseri (C. koseri) account for a smaller number of infections. These pathogens are transported into the urinary tract from the colonic biotope into dysbacteriosis. Urine samples were randomly collected from 249 outpatients who were suspected of having UTIs. After genital cleaning, 10 mL of urine specimens were collected in a sterilized bowel. Then, the specimens were centrifuged at 2,000 rpm for 5 min and the residue was aerobically incubated with the broth infusion of brain flasks at 37°C for 24 h and then applied with a sterile ring onto blood agar plates and MacConkey agar (OxoidTM). Out of 249 urine samples, the results proved that there were 176 (70.7%) and 51(20.5%) gram-negative and gram-positive bacteria isolates, respectively. However, the results demonstrated that there were 22 (8.8%) urine samples with no growth. In addition, the results showed that eight various antimicrobials are used to treat C. koseri. In the current study, C. koseri was treated with eight different antimicrobial agents. The antimicrobial resistance rate for 7 isolates against Cefotaxime, Ceftriaxone, Ciprofloxacin, and Levofloxacin was high for 6 (85.71%) isolates. The results indicated that 6 and 5 isolates had 85.71% and 71.42% antimicrobial resistance against Ceftazidime and Levofloxacin, respectively. Whereas Gentamicin showed a moderate rate of resistance (4 isolates, 57.14%), and Amikacin resistance was found in 5 isolates, accounting for 28.57%. The bacterial isolates had a high susceptibility rate to Imipenem. The qnrA gene was found in 6 (85.71%) isolates. However, the recorded data demonstrated that there is no isolate carrying the qnrC gene. Among all pathogenic bacteria, C. koseri was the lowest causative agent of UTI in this study and was highly resistant to most antimicrobials except Imipenem, which was a good antibiotic with 100% sensitivity.

Effect of Antibiotic Therapy in Children with Pyelouretheral Segment Obstruction

The present retrospective study, which was conducted from 2013 to 2018 years, includes data of 131 children with antenatally diagnosed grade III and IV hydronephrosis (SFU). All patients underwent surgical treatment at the age from 3 months to 3 years and did not receive antibiotic prophylaxis. Urinary tract infection was diagnosed in the presence of WBC in urine sample (> 10 WBC/field of view), positive bacterial growth (≥105 CFU/ml) and fever (≥38.5 ° C). Children with duplex system, obstructive megaureter, vesicoureteral reflux, posterior urethral valve, and neurogenic bladder were excluded from this study. Grade III hydronephrosis was detected in 113 children and grade IV in 18 patients. The total incidence of urinary tract infection was 9.2% (12 patients), while in the group of children with grade IV hydronephrosis it was higher (in 3 of 18 - 16.7%) than in children with grade III (in 9 out of 113 - 7.9%) (P <0.05). The incidence of urinary tract infections did not differ significantly depending on the sex or age of the patients. In 7 (58.3%) patients, urinary tract infection was noted before the age of 6 months, with an average age of 2.7 months. A bacteriological study revealed that in eight (66.7%) patients, E. Coli was the causative agent of urinary tract infection. Children with antenatal diagnosed and postnatal confirmed ureteropelvic junction obstruction do not need antibiotic prophylaxis because of the low incidence of urinary tract infection. However, patients with grade IV hydronephrosis under the age of 6 months should be closely follow-up by physicians for the early diagnosis and treatment of urinary tract infection.

Vaginal Inoculation of Uropathogenic Escherichia coli during Estrus Leads to Genital and Renal Colonization.

ABSTRACTUrinary tract infection (UTI) is one of the most prevalent bacterial infections, particularly in women, children, and the elderly. Uropathogenic Escherichia coli (UPEC) is the predominant etiological agent of UTI. Uropathogens are directly instilled in the urinary bladder, bypassing the lower urogenital tract, in the widely used murine model of UTI. We assessed whether vaginal inoculation of UPEC led to UTI and how stages of the estrous cycle would impact bacterial colonization in mice. Mice in proestrus, estrus, metestrus, and diestrus were identified by vaginal cytology and inoculated with UPEC in the vaginal tract. Mice were euthanized 1 day after infection, and bacterial loads in the urogenital tract, liver, and spleen were enumerated. Mice in estrus exhibited the highest and most consistent UPEC burdens in all organs, except the bladder. Vaginal inoculation resulted in bladder colonization in a UPEC strain-specific manner. In contrast, transurethral inoculation of UPEC led to bladder colonization. Importantly, inoculation by both routes led to vaginal and uterine colonization and concomitant systemic dissemination to the spleen and liver. The kinetics of bacterial colonization over 2 weeks following vaginal inoculation was comparable in the urogenital tract. Tissue sections revealed the induction of vaginitis and cystitis upon the vaginal instillation of UPEC. In summary, vaginal inoculation of UPEC in mice during estrus represents a novel approach to investigate infection of the kidneys and genital tract and systemic dissemination from the urogenital tract. Our findings suggest that estrogen primes the urogenital tract to create a conducive milieu for UPEC colonization.

Synergistic Antibacterial Potential of 6-Pentyl-α-pyrone Lactone and Zinc Oxide Nanoparticles against Multidrug-Resistant Enterobacterales Isolated from Urinary Tract Infections in Humans.

Urinary tract infection (UTI) is one of the most common bacterial infections in the world, which is associated with high morbidity and mortality rates. Enterobacterales species are considered the most causative agent for UTI, especially uropathogenic Escherichia coli (UPEC). Here, we investigated the antibacterial activity of the green fungal metabolite, 6-pentyl α pyrone lactone, alone or in combination with zinc oxide nanoparticles (ZnONPs) against multidrug-resistant Enterobacterales recovered from UTI. The results revealed that 57.27% of human urine samples were positive for Enterobacterales, where E. coli was the most prevalent bacterial pathogen (66.67%). Of note, 98.41% of Enterobacterales isolates were multidrug-resistant (MDR) with multiple antimicrobial resistance (MAR) indices ranged from 0.437 to 1. Fifty percent of the examined isolates were positive for the integrase gene; 60% out of them harbored class 2 integron, whereas the other 40% carried class 1 integrons. The broth microdilution assay ensured that the 6-pentyl-α-pyrone lactone had a reasonable antimicrobial effect against the examined isolates (Minimum inhibitory concentration (MIC) values of 16–32 μg/mL). However, ZnONPs showed a strong antimicrobial effect against the investigated isolates with MIC values ranging from 0.015 to 32 μg/mL. Interestingly, the MICs decreased 5–12 fold and 3–11 fold for 6-pentyl-α-pyrone lactone and ZnONPs, respectively, against examined isolates after their combination. This is the first report suggesting the use of 6-pentyl α pyrone lactone and ZnONPs combination as a promising candidate against MDR Enterobacterales recovered from UTI.

Prevalence of E. coli as a causative agent of urinary tract infections and its drug susceptibility patterns among pregnant mothers seeking medicare at Kisii teaching and referral hospital, Kenya

Background: E-coli, a gram-negative entero-bacteria has been associated with urinary tract infections (UTIs) and antimicrobial resistances in human and animals. This study aimed at establishing the prevalence of E. coli among other bacteria causing UTIs in pregnant women seeking Medicare at Kisii teaching and referral hospital (KTRH), Kenya and to establish the drug susceptibility patterns of the isolated E. coli. Setting-The project was done at Kisii teaching and referral hospital, Kenya.Methods: This hospital based experimental and cross-sectional study conducted in 3 months between March and June 2020 involved 119 pregnant women whose urine samples were cultured on Cysteine Leucine Electrolyte deficiency media (CLED) at 370C overnight and sub-cultured on Mueller Hinton media. Bacterial identification was done by Gram stain and biochemical characterization using indole, methyl-red, Voges-Proskaur and citrate tests while susceptibility tests were conducted by Kirby Bauer disc diffusion technique.Results: Out of the 119 urine samples, E. coli 28 (23.5%) was the second most prevalent after S. aureus 40 (33.6%). Others included S. epidermidis 27 (22.7%), and Proteus spp. 9 (7.6%). All E. coli isolates, 28 (100%) demonstrated resistance to sulfamethoxazole followed by amoxyclave 24 (85.75%), and ceftriaxone 20 (71.42%). They were least resistant to gentamycin 4 (14. 28%) and ofloxacin 6 (21.42%).Conclusions: E. coli which largely exists as a commensal can cause UTIs and could be possessing antimicrobial resistant genes responsible for treatment failure. This demands for new effective therapeutic alternatives and more research on bacterial drug resistant.

"Omics" Technologies - What Have They Told Us About Uropathogenic Escherichia coli Fitness and Virulence During Urinary Tract Infection?

Uropathogenic Escherichia coli (UPEC) is the main etiological agent of urinary tract infection (UTI), a widespread infectious disease of great impact on human health. This is further emphasized by the rapidly increase in antimicrobial resistance in UPEC, which compromises UTI treatment. UPEC biology is highly complex since uropathogens must adopt extracellular and intracellular lifestyles and adapt to different niches in the host. In this context, the implementation of forefront ‘omics’ technologies has provided substantial insight into the understanding of UPEC pathogenesis, which has opened the doors for new therapeutics and prophylactics discovery programs. Thus, ‘omics’ technologies applied to studies of UPEC during UTI, or in models of UTI, have revealed extensive lists of factors that are important for the ability of UPEC to cause disease. The multitude of large ‘omics’ datasets that have been generated calls for scrutinized analysis of specific factors that may be of interest for further development of novel treatment strategies. In this review, we describe main UPEC determinants involved in UTI as estimated by ‘omics’ studies, and we compare prediction of factors across the different ‘omics’ technologies, with a focus on those that have been confirmed to be relevant under UTI-related conditions. We also discuss current challenges and future perspectives regarding analysis of data to provide an overview and better understanding of UPEC mechanisms involved in pathogenesis which should assist in the selection of target sites for future prophylaxis and treatment.

Antibiogram and Frequency of BLA-NDM-1 Gene in E.Coli Isolated from Carbapenem Resistant Cases of UTI Patients

Background: Urinary tract infections (UTI) are very common in all the population around the world. Women are the usual victims of the super bug Escherichia coli (E.coli) and usually suffer from UTI once in a lifetime. The multi-drug resistant (MDR) E.coli is very intelligent creature and turned resistant to the antibiotics through transposons, mutations and plasmids. Most of the E.coli harbors the gene bla-NDM-1 due to which they have become highly resistant to Carbapenem antibiotics. Objectives: To find the bla-NDM-1 gene in Carbapenem resistant E.coli isolated from UTI patients. To determine the frequency of Carbapenem resistant E.coli in patients attending tertiary care hospital of Peshawar. Methodology: This study was conducted in Khyber Teaching Hospital Peshawar from January 2019 to June 2019. we collected 87 urinary E.coli isolates which were resistant to Carbapenem by disc diffusion method on Mueller Hinton Agar. Polymerase chain reaction (PCR) was done to unyield the presence of bla-NDM-1 gene in these 87 cases. Results: According to this study E.coli was the most common causative agent for UTI and females suffered more than male patients. The antibiotics Tigecycline and Colistin showed 100% sensitivity against E.coli. Some of the antibiotics like Ampicillin; Meropnem etc were 100% resistant to E.coli. The gene bla-NDM-1 turned out to be positive in 26.43% of the cases. Conclusion: This study concluded that all the E.coli isolated from UTI patients having bla-NDM-1 gene exhibit resistance to Carbapenem. These isolates are very difficult to treat and limited therapeutic options are available. Keywords: Multi- drug resistant Escherichia coli, Carbapenemases, Antibiogram, Plasmids, Polymerase chain reaction, Transposons and Mutations.

An Overview: Antibiotic Sensitive Pattern in Urinary Tract Infection

Urinary tract infections (UTIs) are the inflammatory disorders of the urinary tract caused by the abnormal growth of pathogens. Urinary tract infections can be community acquired or nosocomial. Symptoms of UTIs such as fever, burning sensations while urinating, LAP, itching, formation of blisters and ulcers in the genital area, genital and supra pubic pain, and pyuria generally depend on the age of the person infected the location of the urinary tract infected. E.coli is the major etiologic agent in causing UTI, which accounts for up to 75%-90% of cases whereas Staphylococcus saprophyticus causes an estimated 5 - 1 5% of UTIs frequently in younger women. P. mirabilis, Klebsiella species, P. aeruginosa and Enterobacter species are less frequent offenders. Less commonly, Enterococci, G. vaginalis and U. urealyticum are also known agents in UTIs. Due to the rapidly evolving adaptive strategies of bacteria, the etiology of UTI and antibiotic resistance profile of bacterial uropathogens have changed considerably over the past years, both in community and nosocomial infections. Treatment of UTIs cases is often started empirically and therapy is based on information determined from the antimicrobial resistance pattern of the urinary pathogens.

Alkalising agents in urinary tract infections: theoretical contraindications, interactions and synergy.

Introduction:Alkalising agents have the potential to enhance the efficacy of many antimicrobial agents used in the treatment of Urinary Tract Infections; they also have the potential to cause significant patient harm if used incorrectly. This work seeks to illustrate and quantify these risks and synergies by modelling drug solubility and supersaturation against pharmacokinetic data for commonly used antibiotic agents.Methods:Solubility-pH relationships are employed to quantify the crystalluria risk for compounds which may be reasonably expected to be co-prescribed—or co-administered—with urinary alkalisers (amoxicillin, nitrofurantoin, trimethoprim, sulfamethoxazole and ciprofloxacin). These results are correlated against reports of crystalluria in the literature and in the EU Adverse Drug Reaction database.Results and Discussion:We find a correlation between the maximum theoretical supersaturation attainable and crystalluria reports for sulfamethoxazole, amoxicillin and ciprofloxacin. Shifts in urine pH which can be induced by alkalising agents may produce supersaturated states (and thus induce crystalluria) and may also affect antimicrobial efficacy. The importance of employing biorelevant media to improve predictive capacity of this analysis is also discussed.Conclusion:Despite their widespread use, alkalising agents have significant effects on the pharmacokinetics of the most common drugs used to treat UTIs. With self-care set to increase, all OTC products should be critically re-evaluated to ensure patient safety, particularly within contexts where healthcare professionals are not involved in treatment selection. This analysis suggests a need for consistency across patient and healthcare professional documents to improve clarity. Plain Language Summary OTC Alkalising agents need additional warning information Alkalising agents (e.g., sodium and potassium citrate) can be purchased in many locations without the supervision of a healthcare professional.Although they are thought as innocuous agents, alkalisers can greatly influence the way some antibiotics behave in the body and this can potentially cause patient harm.This work illustrates these risks and synergies by modelling drug solubility and supersaturation against pharmacokinetic data for commonly used antibiotic agents.Manufacturers and patients should be aware that the use of alkalising agents with these drugs (and potentially many others) may cause unintended consequences.

The aggregate-forming pili (AFP) mediates the aggregative adherence of a hybrid-pathogenic Escherichia coli (UPEC/EAEC) isolated from a urinary tract infection

ABSTRACT Enteroaggregative Escherichia coli (EAEC) comprises an important diarrheagenic pathotype, while uropathogenic E. coli (UPEC) is the most important agent of urinary tract infection (UTI). Recently, EAEC virulence factors have been detected in E. coli strains causing UTI, showing the importance of these hybrid-pathogenic strains. Previously, we detected an E. coli strain isolated from UTI (UPEC-46) presenting characteristics of EAEC, e.g., the aggregative adherence (AA) pattern and EAEC-associated genes (aatA, aap, and pet). In this current study, we analyzed the whole genomic sequence of UPEC-46 and characterized some phenotypic traits. The AA phenotype was observed in cell lineages of urinary and intestinal origin. The production of curli, cellulose, bacteriocins, and Pet toxin was detected. Additionally, UPEC-46 was not capable of forming biofilm using different culture media and human urine. The genome sequence analysis showed that this strain belongs to serotype O166:H12, ST10, and phylogroup A, harbors the tet, aadA, and dfrA/sul resistance genes, and is phylogenetically more related to EAEC strains isolated from human feces. UPEC-46 harbors three plasmids. Plasmid p46-1 (~135 kb) carries some EAEC marker genes and those encoding the aggregate-forming pili (AFP) and its regulator (afpR). A mutation in afpA (encoding the AFP major pilin) led to the loss of pilin production and assembly, and notably, a strongly reduced adhesion to epithelial cells. In summary, the genetic background and phenotypic traits analyzed suggest that UPEC-46 is a hybrid strain (UPEC/EAEC) and highlights the importance of AFP adhesin in the adherence to colorectal and bladder cell lines.

GENETIC DIVERSITY OF ENTEROCOCCUS FAECALIS ISOLATES FROM CHILDREN WITH URINARY TRACT INFECTION IN PRIMORSKY KRAI OF RUSSIA

Background: Escherichia coli is the most common cause of urinary tract infections (UTI). However, Enterococcus faecalis has been shown to be the most common causative agent of UTI among children and newborns in Primorsky Krai of Russia warranting further research. Aim: To study the occurrence of pathogenicity factor genes in the E. faecalis isolates from children with UTI in the Primorsky Krai of the Russian Far East. Methods: Forty-two E. faecalis clinical isolates from children under the age of 16 with UTI identified in 2013-2017 were studied. Phylogenetic diversity of the strains was assessed by the ultilocus sequence typing. Six genes, namely, cylA, aggA, efaA, eep, gelE, esp were tested in the isolates by polymerase chain reaction . Results: CylA, aggA, efaA, eep, gelE and esp genes occurred in 50.0 %, 80.95 %, 100 %, 100 %, 76.2 % and 71.4 % of the isolates, respectively. Eleven different gene variants were detected for the combination of pathogenicity factor genes. The most common gene variants were (aggA, cylA, efaA, eep, gelE, esp) and (aggA, efaA, eep, gelE). Among the uropathogenic E. faecalis. Fourteen sequence-types were identified (ST6, ST16, ST21, ST25, ST40, ST41, ST64, ST116, ST133, ST151, ST179, ST480, ST537, ST774), with ST179, ST774, ST6 being the most common. Conclusions: The identified diversity of sequence-types indicates the genetic heterogeneity of uropathogenic enterococci isolated in the Primorsky Krai. The detection of a large amount of pathogenicity factors and their combinations causes the predominance of E. faecalis in the region as a clinically relevant etiological agent of UTI among children. The identification of highly virulent sequence types such as E. faecalis ST6, ST179 and ST774 warrants further research to determine the population structure of enterococci.

Pathogen profile of urinary tract infections in Nephrology Unit

Abstract Introduction. Urinary tract infection (UTI) is one of the most common types of infection in both hospitalized and outpatient settings. The etiology is mostly bacterial, and the typical causative agent is uropathogenic Escherichia coli. There is a noticeable increase in drug resistance of pathogenic microorganisms. The aim of the study was retrospective analyses of etiological agents of UTI and their antibiotic resistance patterns in Nephrology Unit patients. Material and methods. An infection was diagnosed based on the patient’s symptoms and positive results of urine culture, carried out over 26 months. The clinical material was tested by using the VITEK system, the drug susceptibility of the emerged pathogens was identified. Results. The most common etiological agents of UTI were Gram-negative rods: Escherichia coli (51.23%), Klebsiella spp. (19.3%) and Proteus spp. (13.68%). The analysis of drug resistance profiles of these pathogens showed a high percentage of strains resistant to broad-spectrum penicillins and fluoroquinolones. At the same time, it seems that E. coli isolates presented the most favorable pattern of drug susceptibility in this comparison. Conclusions. The alarming tendency of increasing drug resistance among pathogens causing UTIs to antibiotics such as penicillins or fluoroquinolones prompts a careful choice of drugs in empirical therapies. The most appropriate practice in this regard seems to be meticulous control of nosocomial infections and making therapeutic decisions based on the knowledge of local microbiological data.

Uropathogenic Escherichia coli Shows Antibiotic Tolerance and Growth Heterogeneity in an In Vitro Model of Intracellular Infection.

Uropathogenic Escherichia coli (UPEC), the major causative agent of urinary tract infections, can invade different types of host cells. To compare the pharmacodynamic properties of antibiotics against intra- and extracellular UPEC, an in vitro model of intracellular infection was established in J774 mouse macrophages infected by the UPEC strain CFT073. We tested antibiotics commonly prescribed against urinary tract infections (gentamicin, ampicillin, nitrofurantoin, trimethoprim, sulfamethoxazole, and ciprofloxacin) and the investigational fluoroquinolone finafloxacin. The metabolic activity of individual bacteria was assessed by expressing the fluorescent reporter protein TIMERbac within CFT073. Concentration-response experiments revealed that all tested antibiotics were much less effective against intracellular bacteria than extracellular ones. Most antibiotics, except fluoroquinolones, were unable to reach a bactericidal effect intracellularly at clinically achievable concentrations. Ciprofloxacin and finafloxacin killed 99.9% of extracellular bacteria at concentrations around the MIC, while for intracellular bacteria, concentrations more than 100× over the MIC were required to achieve a bactericidal effect. Time-kill curves showed that finafloxacin was more rapidly bactericidal in acidic medium than at neutral pH, while the reverse observation was made for ciprofloxacin. Intracellularly, kill curves showed biphasic kinetics for both fluoroquinolones, suggesting the presence of drug-tolerant subpopulations. Flow cytometry analysis of TIMERbac fluorescence revealed a marked heterogeneity in intracellular growth of individual bacteria, suggesting that the presence of subpopulations reaching a state of metabolic dormancy was the main reason for increased antibiotic tolerance of intracellular UPEC.

Antibacterial activity of Urena lobata against uropathogens

Urinary tract infections (UTI) are one of the most common form of bacterial infections but the treatment becomes cumbersome as the etiological bacteria are developing resistance against antibiotics. This present study evaluated the efficacy of antimicrobial activity of Urena lobata against uropathogens. Six urine samples from UTI patients were collected from Pathological Laboratory, G.B. Vaghani Multispecialty Hospital, Surat. Bacteria were isolated from these samples using Nutrient agar, Mac Conkey agar plate, Blood agar, Mannitol salt agar, Eosin Methylene Blue agar and King’s agar. The bacterial isolates were identified using cultural characteristics, microscopic features and biochemical characteristics. Leaf extract of Urena lobata was prepared using Soxhlet Extraction Method whereby methanol and distilled water were the extractants used. Herbal extract disc was prepared at concentrations of 50,75, and100 mg/ml and tested against all the isolates. DMSO and antibiotics (Nitrofurantion, Amikacin, Levofloxacin, Norofloxacin, Ofloxacin and Cephalosporins) were used as negative and positive controls respectively.Staphylococcus aureus, Escherichia coli, Bacillus cereus, Streptococcus pneumoniae, Klebsiella spp. and Brevibacillus panacihumi were isolated from the urine samples. All concentrations of aqueous and methanolic extracts of U. lobata leaf displayed highest zone of inhibition against B. cereus. No inhibitory effect was observed against the growth of Klebsiella except at the highest concentrations. Further study is encouraged on the in-vivo study of efficacy of U. lobata on etiological agent of UTI.