Agent Of Urinary Tract Infections Research Articles

Draft genome of three uropathogenic Escherichia coli strains.

Uropathogenic Escherichia coli (UPEC) remains the main etiological agent of urinary tract infections affecting females and males. The draft genome sequence of three strains of UPEC isolated from senior citizens and pregnant women in the state of Puebla, Mexico, is reported here.

Competitive fitness of asymptomatic bacteriuria E. coli strain 83972 against uropathogens in human urine.

Urinary tract infection (UTI) is one of the most common bacterial infections worldwide. The main causative agent of UTI is uropathogenic Escherichia coli (UPEC). There is an immediate need for novel prophylactic and treatment strategies against UTI because of the increasing incidence of antimicrobial resistance among uropathogens. ABU 83972, an asymptomatic bacteriuria-causing E. coli strain, prevents UTI by suppressing the colonization of UPEC. However, the nature of competition and growth repression of UPEC by ABU 83972 is unclear and is the subject of our investigation. Here, we characterized the growth kinetics of ABU 83972 and uropathogens in human urine and laboratory media. Next, we performed a series of competitive co-culture experiments where ABU 83972 and uropathogens were inoculated at a 1:1 ratio in human urine and in various media, and their relative abundance was determined. In human urine, ABU 83972 outcompeted UPEC and additional uropathogens, reaching up to 90% of the total population after 24 hours of incubation. In contrast, UPEC outcompeted ABU 83972 in LB and M9 minimal media and exhibited superior colonization than ABU 83972 in the mouse urinary bladder. Since engineered living materials (ELMs) can be used to retain an organism of interest in a particular location, we developed ABU 83972-containing ELMs that effectively outcompeted UPEC in human urine. In summary, our work establishes that ABU 83972 outcompetes UPEC in a milieu- and cell-density-dependent manner, highlighting the importance of the metabolites and nutrients found in the human urine as determinants of the competitive fitness of ABU 83972.

Gentamicin loaded niosomes against intracellular uropathogenic Escherichia coli strains

Urinary tract infections (UTIs) are the most common bacterial infections and uropathogenic Escherichia coli (UPEC) is the main etiological agent of UTIs. UPEC can persist in bladder cells protected by immunological defenses and antibiotics and intracellular behavior leads to difficulty in eradicating the infection. The aim of this paper is to design, prepare and characterize surfactant-based nanocarriers (niosomes) able to entrap antimicrobial drug and potentially to delivery and release antibiotics into UPEC-infected cells. In order to validate the proposed drug delivery system, gentamicin, was chosen as “active model drug” due to its poor cellular penetration. The niosomes physical–chemical characterization was performed combining different techniques: Dynamic Light Scattering Fluorescence Spectroscopy, Transmission Electron Microscopy. Empty and loaded niosomes were characterized in terms of size, ζ-potential, bilayer features and stability. Moreover, Gentamicin entrapped amount was evaluated, and the release study was also carried out. In addition, the effect of empty and loaded niosomes was studied on the invasion ability of UPEC strains in T24 bladder cell monolayers by Gentamicin Protection Assay and Confocal Microscopy. The observed decrease in UPEC invasion rate leads us to hypothesize a release of antibiotic from niosomes inside the cells. The optimization of the proposed drug delivery system could represent a promising strategy to significatively enhance the internalization of antimicrobial drugs.

High prevalence of plasmid-mediated quinolone resistance in escherichia coli strains producing extended-spectrum beta-lactamases isolated from faeces and urine of pregnant women with acute cystitis.

Escherichia coli is the most common etiological agent of urinary tract infections (UTIs). Meanwhile, plasmid-mediated quinolone resistance (PMQR) is reported in E. coli isolates producing extended-spectrum β-lactamases (ESBLs). Furthermore, the reservoirs and mechanisms of acquisition of uropathogenic Escherichia coli (UPEC) strains are poorly understood. On the other hand, UTIs are common in pregnant women and the treatment challenge is alarming. In the present study, 54 pregnant women with acute cystitis were included. A total of 108 E. coli isolates, 54 isolates from UTI and 54 isolates from faeces of pregnant women (same host) were collected. In the antimicrobial susceptibility test, the highest rate of antibiotic resistance was to nalidixic acid (77%, 83/108) and the lowest rate was to imipenem (9%, 10/108). Among the isolates, 44% (48/108) were ESBLs producers. A high frequency of PMQR genes was observed in the isolates. The frequency of PMQR genes qnrS, qnrB, aac(6')-Ib-cr, and qnrA was 58% (63/108), 21% (23/108), 9% (10/108), and 4% (4/108), respectively. Meanwhile, PMQR genes were not detected in 24% (20/85) of isolates resistant to nalidixic acid and/or fluoroquinolone, indicating that other mechanisms, i.e. chromosomal mutations, are involved in resistance to quinolones, which were not detected in the present study. In ESBL-producing isolates, the frequency of PMQR genes was higher than that of non-ESBL-producing isolates (81% vs. 53%). Meanwhile, UTI and faeces isolates mainly belonged to phylogenetic group B2 (36/54, 67% and 25/54, 46%, respectively) compared to other phylogenetic groups. In addition, virulence factors and multidrug-resistant (MDR) were mainly associated with phylogenetic group B2. However, predominant clones in faeces were not found in UTIs. Rep-PCR revealed the presence of 85 clones in patients. Among the clones, 40 clones were detected only in faeces (faeces-only), 35 clones only in UTI (UTI-only) and 10 clones in both faeces and UTI (faeces-UTI). We found that out of 10 faeces-UTI clones, 5 clones were present in the host's faeces flora. This study revealed a high rate of resistance to the quinolone nalidixic acid and a widespread distribution of PMQR genes in MDR E. coli strains producing ESBLs. The strains represented virulence factors and phylogenetic group B2 are closely associated with abundance in UTI and faeces. However, the predominant clones in faeces were not found in UTIs and it is possible that rep-PCR is not sufficiently discriminating clones.

Nucleoside-diphosphate kinase of uropathogenic Escherichia coli inhibits caspase-1-dependent pyroptosis facilitating urinary tract infection

Uropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infection (UTI). UPEC invades bladder epithelial cells (BECs) via fusiform vesicles, escapes into the cytosol, and establishes biofilm-like intracellular bacterial communities (IBCs). Nucleoside-diphosphate kinase (NDK) is secreted by pathogenic bacteria to enhance virulence. However, whether NDK is involved in UPEC pathogenesis remains unclear. Here, we find that the lack of ndk impairs the colonization of UPEC CFT073 in mouse bladders and kidneys owing to the impaired ability of UPEC to form IBCs. Furthermore, we demonstrate that NDK inhibits caspase-1-dependent pyroptosis by consuming extracellular ATP, preventing superficial BEC exfoliation, and promoting IBC formation. UPEC utilizes the reactive oxygen species (ROS) sensor OxyR to indirectly activate the regulator integration host factor, which then directly activates ndk expression in response to intracellular ROS. Here, we reveal a signaling transduction pathway that UPEC employs to inhibit superficial BEC exfoliation, thus facilitating acute UTI.

The inner membrane protein YhiM links copper and CpxAR envelope stress responses in uropathogenic E. coli.

Urinary tract infection (UTI) is a ubiquitous infectious condition, and uropathogenic Escherichia coli (UPEC) is the predominant causative agent of UTI. Copper (Cu) is implicated in innate immunity, including against UPEC. Cu is a trace element utilized as a co-factor, but excess Cu is toxic due to mismetalation of non-cognate proteins. E. coli precisely regulates Cu homeostasis via efflux systems. However, Cu import mechanisms into the bacterial cell are not clear. We hypothesized that Cu import defective mutants would exhibit increased resistance to Cu. This hypothesis was tested in a forward genetic screen with transposon (Tn5) insertion mutants in UPEC strain CFT073, and we identified 32 unique Cu-resistant mutants. Transposon and defined mutants lacking yhiM, which encodes a hypothetical inner membrane protein, were more resistant to Cu than parental strain. Loss of YhiM led to decreased cellular Cu content and increased expression of copA, encoding a Cu efflux pump. The CpxAR envelope stress response system was activated in the ΔyhiM mutant as indicated by increased expression of cpxP. Transcription of yhiM was regulated by CueR and CpxR, and the CpxAR system was essential for increased Cu resistance in the ΔyhiM mutant. Importantly, activation of CpxAR system in the ΔyhiM mutant was independent of NlpE, a known activator of this system. YhiM was required for optimal fitness of UPEC in a mouse model of UTI. Our findings demonstrate that YhiM is a critical mediator of Cu homeostasis and links bacterial adaptation to Cu stress with the CpxAR-dependent envelope stress response in UPEC.IMPORTANCEUPEC is a common bacterial infection. Bacterial pathogens are exposed to host-derived Cu during infection, including UTI. Here, we describe detection of genes involved in Cu homeostasis in UPEC. A UPEC mutant lacking YhiM, a membrane protein, exhibited dramatic increase in resistance to Cu. Our study demonstrates YhiM as a nexus between Cu stress and the CpxAR-dependent envelope stress response system. Importantly, our findings establish NlpE-independent activation of CpxAR system during Cu stress in UPEC. Collectively, YhiM emerges as a critical mediator of Cu homeostasis in UPEC and highlights the interlinked nature of bacterial adaptation to survival during Cu and envelope stress.

Prevalence of bacterial isolates and change in their antibiotic susceptibility patterns in urinary tract infections- A five year retrospective study

Urinary Tract Infection (UTI) continues to be one of the commonly occurring infections in medical practice despite the widespread availability of antibiotics. Presence of risk factors like elderly age, diabetes, immune-suppression, and other comorbidity pose a challenge in its treatment. Over the years, the etiology and antibiotic resistance of the uropathogens has been inconsistent. Therefore, it is important to determine the causative agents of UTI and their resistance patterns locally, in each region, so that appropriate empiric therapy can be started before the culture and antibiotic sensitivity results are available. This study aims to highlight the prevalence of uropathogens and determine the antibiotic sensitivity pattern over the period of five years.The urine samples from patients with suspected urinary tract infections (UTI) received in the laboratory at Nanavati Max Super speciality hospital were processed by standard protocol from January 2018 to December 2022. Antibiotic susceptibility test for the bacterial isolates was carried out by the VITEK2 automated system and interpreted as per CLSI guidelines. The data collected over the study period of five years was interpreted by using the WHO Net Antibiotic Susceptibility Surveillance Software in February 2023. From a total of 8068 urine samples received over five years, uropathogens were isolated in 4209 (52.16%) samples. E.coli was the most common isolate causing UTI, with prevalence of 380(46.7%) in 2018 and 480(50.6%) in 2022 of total samples. Amongst gram positive organisms, a rise in Enterococcus species was seen from 41(5%) in 2018 to 87(9%) in 2022, with significantly fewer Staphylococcus species isolated, from 32(4%) in 2018 to 7(0.7%) in 2022. The Enterobacteriaceae showed high sensitivity to Amikacin, Tigecycline and Meropenem all throughout the study. However, rise in resistance to Nitrofurantoin and Fosfomycin, antibiotics commonly used for UTIs, was observed. Also the prevalence of Extended Spectrum of Beta lactamase (ESBL) producers decreased from 213(36%) to 127(17%) with a rise in Carbapenemase producing Enterobacteriaceae from 48(8%) to 298(40%) over the five years study period.Amongst Lactose Nonfermenting gram negative bacilli only 32 % were sensitive to Carbapenems and 37% sensitive to Aminoglycosides.By 2022, Enterococcus species showed an increase in sensitivity to Nitrofurantoin (27%), High level Aminoglycosides (37%), Fluoroquinolones (25%) and Ampicillin (37%).The change in antibiotic sensitivity patterns over time and rise in resistance to antibiotics commonly used in empirical treatment of UTIs warrants the need for regular surveillance, so the physicians can be updated regarding the most prevalent organism and its susceptibility and the empirical treatment for UTIs can be modified accordingly. This can help curb indiscriminate use of antibiotics and development of antibiotic resistance.

Exosomes derived from bladder epithelial cells infected with uropathogenic Escherichia coli increase the severity of urinary tract infections (UTIs) by impairing macrophage function.

Uropathogenic Escherichia coli (UPEC) is the primary causative agent of urinary tract infections (UTIs) in humans. Moreover, as one of the most common bacterial pathogens, UPEC imposes a substantial burden on healthcare systems worldwide. Epithelial cells and macrophages are two major components of the innate immune system, which play critical roles in defending the bladder against UPEC invasion. Yet, the routes of communication between these cells during UTI pathogenesis are still not fully understood. In the present study, we investigated the role of membrane-bound nanovesicles (exosomes) in the communication between bladder epithelial cells and macrophages during UPEC infection, using an array of techniques such as flow cytometry, miRNA profiling, RNA sequencing, and western blotting. Moreover, our in vitro findings were validated in a mouse model of UPEC-induced cystitis. We found that UPEC infection induced the bladder epithelial MB49 cell line to secrete large numbers of exosomes (MB49-U-Exo), which were efficiently absorbed by macrophages both in vivo and in vitro. Assimilation of MB49-U-Exo induced macrophages to produce proinflammatory cytokines, including tumor necrosis factor (TNF)α. Exposure of macrophages to MB49-U-Exo reduced their phagocytic activity (by downregulating the expression of phagocytosis-related genes) and increased their rate of apoptosis. Mechanistically, we showed that MB49-U-Exo were enriched in miR-18a-5p, which induced TNFα expression in macrophages by targeting PTEN and activating the MAPK/JNK signaling pathway. Moreover, administration of the exosome secretion inhibitor GW4869 or a TNFα-neutralizing antibody alleviated UPEC-mediated tissue damage in mice with UPEC-induced cystitis by reducing the bacterial burden of the bladder and dampening the associated inflammatory response. Collectively, these findings suggest that MB49-U-Exo regulate macrophage function in a way that exacerbates UPEC-mediated tissue impairment. Thus, targeting exosomal -release or TNFα signaling during UPEC infection may represent promising non-antibiotic strategies for treating UTIs.

Fungal Endophytes - An Alternative Source of Bioactive Compounds from Plant Species for Prospective Applications – Review

Plant derived compounds have a strong antagonistic activity on pathogenic microorganisms using tremendous natural bioactive compounds accumulated in different parts of plants. Limonoids are important group of metabolically altered triterpene in Meliaceae. Out of 300 limonoids known today, Melia azedarach (melianone, meliarachin, meliacine), Melia dubia (nimbin, meliacarpin) and Azadirachta indica (azadirachtin, nimbolide) contributes about one-third of limonoids. Endophytic fungi are plant symbionts that act as an elicitor for the production of secondary metabolites which serves as an excellent source of drugs for various ailments. It has a great potential to synthesize several novel bioactive compounds which paved a way for commercial exploitation of secondary metabolites for pharmaceutical and agricultural sectors. Many dominant endophytic fungi isolated from these plants includes Colletotrichum sp, Trichoderma sp, Penicillium citrinum, Fusarium solani, Aspergillus sp, Nigrospora sp and Alternaria sp which acts on human pathogenic microorganisms such as Escherichia coli Pseudomonas aeruginosa, Staphylococcus aureus, Klebsiella pneumoniae and Candida albicans common causative agents for Urinary Tract Infections (UTI) predominantly occurring in women. This review aims to delineate the overview on sources of limonoids present in the leaves and the endophytic fungi isolated from these leaves of Meliaceae. The isolated endophytic fungi from the leaves exhibited antagonistic activity on UTI causing pathogens which often leads to infertility among young women. Thus, the sources of limonoids and isolation of endophytic fungi can be enhanced by reinforcing the production of commercial bioactive secondary metabolites that can be substantially utilized by pharmaceutical and agricultural industries to safeguard human beings.

Urinary tract infections in children with cancer and febrile neutropenia - single center experience

Background/Aim. Urinary tract infection (UTI) in children with febrile neutropenia (FN) after receiving chemotherapy could be followed by atypical symptoms and signs. The absence of routine urine culture (UC) sampling for analysis could lead to undiagnosed disease and inadequate treatment of these patients. The aim of the study was to indicate the importance of sampling UC in children who have developed FN and to point out the most probable causative agents of UTI in children with FN and antibiotic sensitivity/resistance of the isolated strains. Methods. During a five-year observation period, 40 UTIs were registered in 30 patients with FN. In the study group of patients with FN, the number of UTIs, the number of recurrent UTIs, isolated pathogens, their sensitivity to antibiotics, characteristics of urine sediment, the presence of localized symptoms of UTI, and the presence of urosepsis were analyzed. The obtained results were compared with the control group which consisted of children who were healthy prior to hospitalization due to febrile urinary infection. Results. When compared to the control group, significant differences in the presence of symptoms of UTI and urine sediment findings in patients with FN were observed. A higher percentage of resistant strains of Escherichia coli isolated from UC of cancer patients with FN was noted when compared to the control group. Three UTI cases were followed by urosepsis. Conclusion. UC findings are important not only in establishing the diagnosis of UTI and detecting multi-resistant bacterial strains but also in choosing appropriate antibiotics and selecting a subgroup of patients with recurrent UTI who require further monitoring and detecting potential complications in a timely manner.

Bacterial profile of urinary tract infection among pregnant women: a systematic review

Urinary tract infection (UTI) is the infection and colonization of microorganisms in the urinary tract. Women carry a higher risk of being affected by UTI compared to males, especially during pregnancy due to physiological and hormonal changes. Appropriate and adequate management of UTI reduces the incidence of further maternal and fetal complications. Nonetheless, there is a growing concern regarding resistance to antibiotics among the bacteria causing UTIs in pregnant women. To identify the bacterial profile of UTIs during gestation by exploring the previous research conducted on this subject. A systematic online search was performed on scientific databases to obtain articles related to the current subject using search terms, including bacteria, profile, causative agent, sensitivity, resistance, symptomatic, pregnancy, gestation, and pregnant women. The articles of relevance were considered if they were original articles, written in the English language, available for full-text, and reporting bacterial profile of UTI during pregnancy. A total of eight articles were eligible and met the inclusion criteria. The studies were published between 2020 and 2023 and included a total number of 3,702 pregnant females from five different countries. The bacterial profile of UTI among pregnant women including symptomatic UTI, and resistance and sensitivity of bacteria to antibiotics varied between different regions. However, the causative agents of UTI displayed less variation as Escherichia coli and K.pneomniae were the major causative agents of UTI during pregnancy.

Isolation and Identification of Multidrug-Resistant Raoultella terrigena as a Causative Agent of Urinary Tract Infection in Pregnant Women in the South of Libya

Raoutella terrigena is an opportunistic bacteria that rarely cause infections in humans. It is facultative Gram negative, rod shaped, present mostly in water, plants, soil, fish and insects. Two strains of R. terrigena have been isolated during a study searching of the must causative bacteria of urinary tract infection, in pregnant women where it counted about 2% of the total isolates. Susceptibility test has revealed multi drug resistance to about 61.5 % of the used antibiotics. This is the first documented report of such infection in Libya.

Bacterial Uropathogens causing Urinary Tract Infection at Hetauda Hospital

Introduction: Urinary tract infection is the infection of the uroepithelium that mainly affects kidney, ureter, urinary bladder and urethra. The causative agents of urinary tract infections are bacteria and fungi. Among these, bacteria are the most causative uropathogens of urinary tract infections. Objectives: The objective of this study is to isolate, identify, characterized and perform antibiotic sensitivity test from the bacterial isolates. Methods: A descriptive cross-sectional study was conducted from March 2023 to May 2023. Ethical approval was obtained from Institutional Review Committee (IRC), Madan Bhandari Academy of Health Sciences, IRC-32-079. Total of 1120 urine samples were collected during this study. Convenient sampling technique was used for this study. Clean catch or mid-stream urine was collected in a sterile container. All urine samples were submitted to Department of Microbiology for culture. A set of biochemical tests were done for bacterial identification. Antibiotic sensitivity test was performed by Kirby-Bauer disk diffusion method. Results: Total of 1120 urine samples were collected from patients suspected to Urinary Tract Infections. Total urine culture positive was 340 (30.36%). Of these bacterial isolates, Escherichia coli (248, 73%) was the most common species, followed by Klebsiella species (26, 7.6%), Staphylococcus saprophyticus (23, 6.76%) and Enterococcus fecalis (18, 5.3%). Conclusion: As prevalence of bacteria causing urinary tract infections is increasing day by day; so its proper isolation, identification, characterization with their antibiotic profile is necessary as it forms the base line for the clinicians to choose appropriate antimicrobial agents for empirical as well as rational treatment for UTIs.

Bacterial resistance of the most locally isolated pathogens to commonly used antibiotics in Kerbala/Iraq

Antibiotics are one of the biggest threats to human health. Antibiotics remain the most critical resource in the worldwide management of infections. Identifying the causative agent in an infectious disease is vital to determine their antimicrobial susceptibility. The susceptibility of pathogenic microbes has a critical role in deciding the morbidity and mortality rates of these diseases—the aim of the current study is to investigate the antimicrobial susceptibility of common pathogens in Karbala. The specimens were collected from 370 patients diagnosed with bacterial infections from Al-Hindiya Hospital in 2022. Standard microbiological techniques were implemented. Identification of microorganisms was made using biochemical tests. A pure culture was prepared for antimicrobial susceptibility testing. Antibiotic sensitivity testing was performed using the Kirby-Bauer disk diffusion method, and the results were interpreted. Using the standard operating procedure and the standard bacteriological procedures, we found that the most common pathological microorganisms predominance was Escherichia coli, Staphylococcus aureus, Pseudomonas aeruginosa, and Streptococcus spp. And Shigella spp. The antimicrobial patterns were: Escherichia coli strains were found to be highly sensitive to meropenem (84%) with excellent sensitivity to Levofloxacin (68%) and Nitrofurantoin (64 %) while it highly resistant to Amoxicillin, Erythromycin, Vancomycin, Novobiocin, Trimethoprim-sulfamethoxazole, Cefotaxime, Cefepime and Tetracycline and moderate to Ciprofloxacin, Azithromycin, and Gentamicin. The most common causative agent of urinary tract infections (which are the most common infections in the population studied) was Escherichia coli. Meropenem was a broad-spectrum antimicrobial that many bacterial spp were susceptible to.

The prevalence of urinary tract disease in the elderly,Bani waleed, Libya

Urinary tract infection (UTI) is common infections occurred in elderly. The study aimed to evaluate the prevalence of the UTI in elderly people and to determine the most risk factors for serious clinical complications of UTI.
 Methodology: Data of a descriptive cross sectional study were collected from Central Bani Waleed Hospital, 106 UTI patients over 60 year old were involved in this study.
 Results: The most age group effected with UTI was 70-79 y (35.8%), followed by age group 60-69 y (31.1%). The men old people were more susceptible to UTI than women one (51.8% and 48.1% respectively), as well as elderly diabetic patients were susceptible to UTI more than non-diabetic (58.4% and 41.5% respectively). E.coli represented the most causative agent of UTI (71.7%). Cystitis (65.09 %) was the most prevalent UTI among elderly than pyelonephritis (34.91%).
 Conclusion: As the human age progress , the resistance of immune system deficiencies gradually become more susceptible to infectious diseases. The UTI one of these infections that can lead to serious complications in elderly immunocompromised. In addition, the comorbidities factors increase the directly affected of old people to UTI.

Investigation of Virulence Factors, Phylogenetic Group Features, and the Presence of ST131 Clone in Escherichia coli Isolates, a Urinary Tract Infection Agent in Children

Urinary tract infection (UTI) caused by Escherichia coli is a significant health issue in children. Today especially E.coli O25b/ST131, defined as a pandemic clone, is a serious public health problem due to its high virulence and antimicrobial resistance rates. In this study, a total of 200 (100 first and 100 recurrent UTI-causing) E.coli isolates from urine samples sent to the Ankara University School of Medicine Cebeci Training and Research Hospital Central Laboratory between January and September 2021 with the preliminary diagnosis of UTI in pediatric patients aged three to 18 years were analyzed for antimicrobial resistance rates, phylogenetic group distributions, virulence factor frequencies and whether they belong to the O25b/ST131 clone. It is aimed in this study that, the obtained data will shed light on new studies for diagnosis, treatment and prophylaxis options that can be developed for more effective UTI management by contributing to the surveillance studies in our country. Antimicrobial susceptibility of E.coli isolates identified by conventional methods was evaluated by Kirby-Bauer disc diffusion method and extended spectrum beta-lactamase (ESBL) production was evaluated by double disc synergy test. Polymerase chain reaction (PCR) was used for the investigation of phylogenetic grouping, the O25b/ST131 clone, virulence genes and the molecular level classification of the isolates detected as uropathogenic E.coli (UPEC). Pulsed-field gel electrophoresis (PFGE) was performed with the isolates collected at different times from the same patient. The highest antimicrobial resistance rates observed were against ampicillin (n= 100, 50%), cefazolin (n= 99, 49.5%), trimethoprim-sulfamethoxazole (n= 55, 27.5%), amoxicillin-clavulanic acid (n= 43, 21.5%) and cefotaxime (n= 43, 21.5%). In recurrent UTI agents, resistance rates were higher for cefotaxime (n= 29, 29%), trimethoprim-sulfamethoxazole (n= 35, 35%) and cefepime (n= 25, 25%) and in O25b/ST131 isolates (n= 67) the rates were higher for amikacin (n= 3, 4.5%), gentamicin (n= 10, 14.9%) and ciprofloxacin (n= 17, 25.4%) when compared to the first UTI agents and non-O25b/ ST131 isolates (p< 0.05). It was found that 29% (n = 58) of the isolates were multidrug resistant (MDR) and 19% (n = 38) produced ESBL.The rate of recurrent UTI agents was found to be higher among ESBL producing isolates and/or MDR isolates (n= 36, 62% and n= 27, 71%, respectively, p< 0.05). It was found that 45.5% (n= 91) of the isolates were in D, 37.5% (n= 75) in B2, 12.5% (n= 25) in A, and 4.5% (n= 9) in B1 phylogenetic groups and isolates belonging to B2 and D phylogenetic groups had higher antibiotic resistance rates and carried more virulence genes (p< 0.05). Of the isolates, 33.5% (n= 67) were found to belong to the O25b/ST131 clone, no significant difference was found between the O25b/ST131 rates among the first and recurrent UTI agents (p> 0.05). It was determined that the isolates most frequently carry virulence genes for adhesion [fimH 97% (n= 194), papA 57% (n= 114), yfcV 49.5% (n= 99)] and iron uptake systems [fyuA 85.5% (n= 171), chuA 78% (n= 156), iutA 73% (n= 146)]. All virulence factors were detected more frequently in isolates belonging to the O25b/ST131 clone (p< 0.05). Of the isolates, 97% (n= 65) belonging to the O25b/ST131 clone and 27.1% (n= 36) not belonging to this clone were defined as UPEC with molecular analysis (p< 0.0001). Thirty-three isolates belonging to 15 patients were evaluated with PFGE, and it was observed that the latter isolate and the first isolate of eight patients (53%) had the same band profile. Focusing on surveillance, diagnostic testing, treatment algorithms, and preventive measures for E.coli and especially for ST131 clone, which is frequently observed as causative agent in childhood UTIs, will help to manage challenging E.coli infections.

İdrar Kültürlerinden İzole Edilen Escherichia Coli Suşlarının Antibiyogramlarının Değerlendirilmesi

Objective: Our aim in this study is to determine the antimicrobial resistance profiles of Escherichia coli strains known to cause urinary tract infections most frequently in the region, to obtain microbiological data about their current status, and to contribute to the development of clinical strategies for the prevention and management of these infections.&#x0D; Materials and methods: Our study includes patients with E. coli isolated from urine cultures. In the study, urine cultures and antibiograms of patients between January 2019 and January 2020 were scanned retrospectively. 662 patients over the age of 18 were included in the study. The files of the patients included in the study, the information registered in the system and the pre-filled forms were examined by examining age, gender, reproductive status in urine cultures, ESBL status, antibiograms, hospitalization status, admission outpatient clinic, reason for hospitalization, chronic and additional diseases, symptom status, the blood, nitrite, leukocyte parameters in the urinary dipstick test and the antibiotic treatment started were recorded.&#x0D; Results: The antibiotics most frequently resistant to E. coli strains were cefuroxime (88.5%), tobramycin (75%), ceftazidime (72.9%), cefepime (59.5%) and ampicillin (57.5%). The antibiotics with the highest sensitivity were imipenem (100%), amikacin (99.1%), meropenem (98.2%), nitrofurantoin (96.3%) and fosfomycin (96.3%). It was observed that 25.7% of E. coli strains were ESBL positive.&#x0D; Conclusion: Our findings showed that E. coli strains, which are the most common causative agents of urinary tract infections, have high resistance to many antibiotics commonly used in treatment. Considering the local resistance rates in empirical treatment and starting the treatment by making a urine culture will guide the treatment. As a result of our research, we think that the use of fosfomycin and nitrofurantoin in uncomplicated infections and the use of carbapenems and aminoglycoside antibiotics in complicated infections and upper urinary tract involvement are appropriate in the empirical treatment of urinary tract infections.

Multidrug-resistant E. coli encoding high genetic diversity in carbohydrate metabolism genes displace commensal E. coli from the intestinal tract.

Extra-intestinal pathogenic Escherichia coli (ExPEC) can cause a variety of infections outside of the intestine and are a major causative agent of urinary tract infections. Treatment of these infections is increasingly frustrated by antimicrobial resistance (AMR) diminishing the number of effective therapies available to clinicians. Incidence of multidrug resistance (MDR) is not uniform across the phylogenetic spectrum of E. coli. Instead, AMR is concentrated in select lineages, such as ST131, which are MDR pandemic clones that have spread AMR globally. Using a gnotobiotic mouse model, we demonstrate that an MDR E. coli ST131 is capable of out-competing and displacing non-MDR E. coli from the gut in vivo. This is achieved in the absence of antibiotic treatment mediating a selective advantage. In mice colonised with non-MDR E. coli strains, challenge with MDR E. coli either by oral gavage or co-housing with MDR E. coli colonised mice results in displacement and dominant intestinal colonisation by MDR E. coli ST131. To investigate the genetic basis of this superior gut colonisation ability by MDR E. coli, we assayed the metabolic capabilities of our strains using a Biolog phenotypic microarray revealing altered carbon metabolism. Functional pangenomic analysis of 19,571 E. coli genomes revealed that carriage of AMR genes is associated with increased diversity in carbohydrate metabolism genes. The data presented here demonstrate that independent of antibiotic selective pressures, MDR E. coli display a competitive advantage to colonise the mammalian gut and points to a vital role of metabolism in the evolution and success of MDR lineages of E. coli via carriage and spread.

Green synthesis of silver nanoparticles derived from lemon and pomegranate peel extracts to combat multidrug-resistant bacterial isolates

BackgroundMultidrug-resistant (MDR) bacteria are acknowledged as one of the main factors contributing to chronic illnesses and fatalities globally. Numerous diseases, including bloodstream infections, pneumonia, urinary tract infections, and surgical site infections, can be brought on by MDR bacteria. Therefore, a crucial topic of continuing research is the development of a novel and different treatment for MDR microbial pathogens. This work is introduce an alternative method for elimination of MDR bacterial isolates which are causative agents of urinary tract infection among people in Egypt. In our study, we need a novel strategy to combat MDR bacteria by green-synthesized metal nanoparticles (MNPs). That is due to the ability of MNPs to penetrate the cell wall and the cell membrane of gram-positive and gram-negative bacteria. MethodsClinical isolates of MDR bacteria had their antibiotic susceptibility assessed before being molecularly identified using 16 s rRNA, sequencing, and phylogenetic analysis. Also, genetic profiles of isolated strains were performed using ISSR and SDS-PAGE. Finally, characterized plant-mediated silver nanoparticles derived from lemon and pomegranate peel extracts were evaluated against isolated multidrug-resistant bacterial stains. ResultsIn our present trial, one-hundred urine samples were collected from 71 females and 29 males complaining of UTI (urinary tract infection) symptoms. One-hundred microbial isolates were isolated, including 88-g negative and only 8-g positive bacteria in addition to four yeast isolates (Candida species). A total of 72% of the isolated bacteria showed MDR activity. The most prevalent MDR bacterial isolates (Escherichia coli, Pseudomonas aeruginosa, Acinetobacter baumannii, Enterococcus faecalis, and Klebsiella pneumoniae) were identified through 16S rDNA PCR sequencing as with accession numbers OP741103, OP741104, OP741105, OP741106, and OP741107, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were characterized by UV spectroscopy, FTIR, XRD, and TEM with average size 32 and 28 nm, respectively. Lemon and pomegranate-mediated silver nanoparticles [Ag-NPs] showed an inhibitory effect on the selected five MDR isolates at MIC 50 and 30 µg/mL, respectively. These common bacterial isolates were also genetically examined using ISSR PCR, and their total protein level was evaluated using SDS-PAGE, showing the presence of distinct genetic and protein bands for each bacterial species and emphasizing their general and protein composition as a crucial and essential tool in understanding and overcoming MDR behavior in UTI patients. ConclusionsLemon and pomegranate-mediated silver nanoparticles [Ag-NPs] were found to have an inhibitory effect on MDR isolates. Therefore, the study suggests that [Ag-NPs] could be a potential treatment for MDR UTI infections caused by the identified bacterial species.

Virulence-associated genes and antimicrobial resistance patterns in bacteria isolated from pregnant and nonpregnant women with urinary tract infections: the risk of neonatal sepsis.

Uropathogenic Escherichia coli (UPEC) is classified as the major causative agent of urinary tract infections (UTIs). UPEC virulence and antibiotic resistance can lead to complications in pregnant women and (or) newborns. Therefore, the aim of this study was to determine the etiological agents of UTIs, as well as to identify genes related to virulence factors in bacteria isolated from pregnant and nonpregnant women. A total of 4506 urine samples were collected from pregnant and nonpregnant women. Urine cultures were performed, and PCR was used to identify phylogroups and virulence-related genes. Antibiotic resistance profiles were determined. The incidence of UTIs was 6.9% (pregnant women, n=206 and nonpregnant women, n=57), and UPEC belonging to phylogroup A was the most prevalent. The presence of genes related to capsular protection, adhesins, iron acquisition, and serum protection in UPEC was associated with not being pregnant, while the presence of genes related to adhesins was associated with pregnancy. Bacteria isolated from nonpregnant women were more resistant to antibiotics; 36.5% were multidrug resistant, and 34.9% were extensively drug resistant. Finally, UTIs were associated with neonatal sepsis risk, particularly in pregnant women who underwent cesarean section while having a UTI caused by E. coli. In conclusion, UPEC isolated from nonpregnant women carried more virulence factors than those isolated from pregnant women, and maternal UTIs were associated with neonatal sepsis risk.