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AIMS: Pyometra and cystitis caused by Escherichia coli are common diseases identified in canine or feline females. The origin of pyometra infection remains uncertain, and effective prevention strategies for this disease are still unknown. This study aimed to provide a phenotypic characterization, including antimicrobial resistance and virulence profiles, of endometrial pathogenic (EnPEC) and uropathogenic (UPEC) E. coli strains isolated simultaneously from the same animal. METHODS AND RESULTS: Sixteen E. coli strains, from eight different animals, were analyzed in this study. The antimicrobial susceptibility profile of EnPEC and UPEC strains was determined using the disc diffusion method, which showed a similar susceptibility profile among strains (EnPEC and UPEC) from the same animal. The virulence profile of the strains was assessed through biofilm formation, as well as serum resistance abilities. EnPEC and UPEC strains from the same animal exhibited slight variations in their virulence and antimicrobial resistance capabilities. Overall, most of the strain pairs showed a high similarity in their ability to establish biofilms and survive in serum complement activity. CONCLUSIONS: Overall, strains of E. coli isolated from both pyometra and cystitis in the same animal, despite presenting distinct clinical diseases, exhibit a wide phenotypic similarity, suggesting a common origin for the strains.
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Biopelículas , Enfermedades de los Gatos , Cistitis , Infecciones por Escherichia coli , Escherichia coli , Pruebas de Sensibilidad Microbiana , Fenotipo , Piómetra , Animales , Cistitis/microbiología , Cistitis/veterinaria , Piómetra/microbiología , Piómetra/veterinaria , Femenino , Gatos , Perros , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/veterinaria , Escherichia coli/aislamiento & purificación , Escherichia coli/patogenicidad , Enfermedades de los Gatos/microbiología , Biopelículas/crecimiento & desarrollo , Virulencia , Antibacterianos/farmacología , Enfermedades de los Perros/microbiología , Escherichia coli Uropatógena/aislamiento & purificación , Escherichia coli Uropatógena/patogenicidad , Farmacorresistencia BacterianaRESUMEN
Urinary tract infections (UTIs) are among the most common outpatient infections, with a lifetime incidence of around 60% in women. We analysed urine samples from 223 patients with community-acquired UTIs and report the presence of the cleavage product released during the synthesis of colibactin, a bacterial genotoxin, in 55 of the samples examined. Uropathogenic Escherichia coli strains isolated from these patients, as well as the archetypal E. coli strain UTI89, were found to produce colibactin. In a murine model of UTI, the machinery producing colibactin was expressed during the early hours of the infection, when intracellular bacterial communities form. We observed extensive DNA damage both in umbrella and bladder progenitor cells. To the best of our knowledge this is the first report of colibactin production in UTIs in humans and its genotoxicity in bladder cells.
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Daño del ADN , Infecciones por Escherichia coli/patología , Péptidos/metabolismo , Policétidos/metabolismo , Vejiga Urinaria/patología , Infecciones Urinarias/patología , Escherichia coli Uropatógena/aislamiento & purificación , Anciano , Animales , Infecciones por Escherichia coli/genética , Infecciones por Escherichia coli/microbiología , Femenino , Humanos , Masculino , Ratones , Ratones Endogámicos C3H , Mutágenos/metabolismo , Vejiga Urinaria/metabolismo , Vejiga Urinaria/microbiología , Infecciones Urinarias/genética , Infecciones Urinarias/microbiologíaRESUMEN
Urinary tract infections (UTIs) caused by uropathogenic Escherichia coli (UPEC) affect 150 million people annually. Despite effective antibiotic therapy, 30-50% of patients experience recurrent UTIs. In addition, the growing prevalence of UPEC that are resistant to last-line antibiotic treatments, and more recently to carbapenems and colistin, make UTI a prime example of the antibiotic-resistance crisis and emphasize the need for new approaches to treat and prevent bacterial infections. UPEC strains establish reservoirs in the gut from which they are shed in the faeces, and can colonize the periurethral area or vagina and subsequently ascend through the urethra to the urinary tract, where they cause UTIs. UPEC isolates encode up to 16 distinct chaperone-usher pathway pili, and each pilus type may enable colonization of a habitat in the host or environment. For example, the type 1 pilus adhesin FimH binds mannose on the bladder surface, and mediates colonization of the bladder. However, little is known about the mechanisms underlying UPEC persistence in the gut. Here, using a mouse model, we show that F17-like and type 1 pili promote intestinal colonization and show distinct binding to epithelial cells distributed along colonic crypts. Phylogenomic and structural analyses reveal that F17-like pili are closely related to pilus types carried by intestinal pathogens, but are restricted to extra-intestinal pathogenic E. coli. Moreover, we show that targeting FimH with M4284, a high-affinity inhibitory mannoside, reduces intestinal colonization of genetically diverse UPEC isolates, while simultaneously treating UTI, without notably disrupting the structural configuration of the gut microbiota. By selectively depleting intestinal UPEC reservoirs, mannosides could markedly reduce the rate of UTIs and recurrent UTIs.
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Proteínas Fimbrias/antagonistas & inhibidores , Intestinos/efectos de los fármacos , Intestinos/microbiología , Manósidos/farmacología , Ácidos Ftálicos/farmacología , Infecciones Urinarias/prevención & control , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/aislamiento & purificación , Adhesinas de Escherichia coli/metabolismo , Secuencia de Aminoácidos , Animales , Células Epiteliales/efectos de los fármacos , Células Epiteliales/microbiología , Heces/microbiología , Femenino , Proteínas Fimbrias/metabolismo , Fimbrias Bacterianas/clasificación , Fimbrias Bacterianas/efectos de los fármacos , Fimbrias Bacterianas/genética , Fimbrias Bacterianas/metabolismo , Humanos , Intestinos/citología , Manósidos/uso terapéutico , Ratones , Modelos Moleculares , Ácidos Ftálicos/uso terapéutico , Vejiga Urinaria/efectos de los fármacos , Vejiga Urinaria/microbiología , Infecciones Urinarias/tratamiento farmacológico , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/genéticaRESUMEN
BACKGROUND: Urinary Tract Infection (UTI) is one of the most common bacterial infectious diseases which causes considerable morbidity and costly health problems. Uropathogenic Escherichia coli (UPEC), the most common pathogen causing UTI, is a highly heterogeneous group of extraintestinal pathogenic E. coli (ExPEC) which may carry a variety of virulence factors and belonging to different phylogenetic backgrounds. The current study aimed to investigate the frequency and association between various virulence factors (VFs) and phylogenetic groups of UPEC and commensal isolates. METHODS: UPEC and commensal E. coli strains isolated from UTI and feces of healthy humans were compared for the presence of VFs and phylogenetic groups. Association between virulence genes was investigated and cluster analysis was employed. RESULTS: According to the results, among a 30 virulence markers tested, the pathogenicity-associated island (PAI), papAH, papEF, fimH, fyuA, and traT genes prevalence were statistically significant in UPEC isolates. A strong association was found between the B2 and D phylogenetic groups and clinical isolates of UPEC; while, commensal isolates were mostly associated with phylogenetic group A. The aggregated VFs scores were more than twice higher in the UPEC isolates in comparison with the commensal isolates. Interestingly, the B2 group in both UPEC and commensal isolates had the highest VF scores. A strong positive association was found between several virulence genes. The clustering results demonstrated that UPEC or commensal E. coli isolates were highly heterogeneous due to different composition of their virulence gene pool and pathogenicity islands. CONCLUSION: Genetic structure and VFs of UPEC strains vary from region to region; therefore, to control the UTI, the epidemiological aspects and characterization of the UPEC isolates need to be investigated in different regions. Since UPEC isolates are generally originate from the commensal strains, it may be feasible to reduce the UTI burden by interfering the intestinal colonization, particularly in the highly pathogenic clonal lineages such as B2.
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Infecciones por Escherichia coli/microbiología , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena , Factores de Virulencia/genética , Virulencia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Estudios de Casos y Controles , Niño , Preescolar , Infecciones por Escherichia coli/epidemiología , Proteínas de Escherichia coli/genética , Escherichia coli Patógena Extraintestinal/genética , Escherichia coli Patógena Extraintestinal/aislamiento & purificación , Escherichia coli Patógena Extraintestinal/patogenicidad , Femenino , Islas Genómicas/genética , Humanos , Lactante , Recién Nacido , Irán/epidemiología , Masculino , Persona de Mediana Edad , Filogenia , Prevalencia , Infecciones Urinarias/epidemiología , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/aislamiento & purificación , Escherichia coli Uropatógena/patogenicidad , Adulto JovenRESUMEN
BACKGROUND: This study aimed to investigate the phylogenetic characterization and virulence traits of uropathogenic Escherichia coli (UPEC) isolated from kidney transplant patients (KTPs) as well as non-KTPs and analyze the clonal distribution of Extended spectrum ß-lactamases (ESBLs)-producing UPEC containing blaCTX-M gene. METHODS: To this end, we determined virulence marker and the phylogenetic characterization of UPEC in non-KTPs (n = 65) and KTPs (n = 46). The non-KTPs were considered the control group of the study. Also, according to the Achtman scheme, we performed multilocus sequence typing to assess the relationship between twenty-nine of ESBL-producing isolates containing blaCTX-M gene. RESULTS: According to the results of PCR assay, the prevalence of virulence factor genes ranged from 0% (cnf and papG III) to 93.7% (fimH). Also, KTP isolates significantly differed from non-KTP isolates only in terms of the prevalence of pap GI elements. Moreover, the most frequent UPEC isolates were in phylogenetic group B2, followed by group D (18.9%), and group A (13.5%). Furthermore, except for phylogenetic group C, there was no significant correlation between phylogenetic distribution in KTPs and non-KTPs. Additionally, MLST analysis of blaCTX-M carrying isolates identified 18 unique sequence types (ST) the most common of which was ST131 (24.1%), followed by ST1193 (10.3%), while fourteen STs were detected only once. CONCLUSIONS: The results further revealed significant differences between the UPEC isolates from KTPs and non-KTPs regarding the phylogroups C and PAI gene. Based on MLST analysis, we also observed a relatively high diversity in UPEC isolates obtained from KTPs and non-KTPs. Moreover, clonal complex (CC) 131 and ST131 were found to be the most prevalent clones and ST types, respectively. Besides, for the first time, ST8503 were reported in KTPs. These results suggested regular studies on characterization of UPEC isolates among KTPs.
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Infecciones por Escherichia coli/microbiología , Trasplante de Riñón , Infecciones Urinarias/diagnóstico , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/aislamiento & purificación , Infecciones por Escherichia coli/diagnóstico , Humanos , Irán/epidemiología , Trasplante de Riñón/efectos adversos , Epidemiología Molecular , Tipificación de Secuencias Multilocus , Filogenia , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/enzimología , Virulencia , Factores de Virulencia , beta-Lactamasas/genéticaRESUMEN
BACKGROUND: Uropathogenic Escherichia coli (UPEC) is a major cause of urinary tract infection (UTI); however, treatment of UTI has been challenging due to increased antimicrobial resistance (AMR). One of the most important types of AMR is carbapenem resistance (CR). CR bacteria are known as an important threat to global public health today. Class B metallo-beta-lactamases (MBLs) are one of the major factors for resistance against carbapenems. We aimed to investigate the characteristics of UPEC isolates producing MBL. METHODS: A cross-sectional study was conducted from October 2018 to December 2019 in Ahvaz; Iran. UPEC isolates were identified by biochemical and molecular methods. Metallo-beta-lactamase-producing isolates were detected using modified carbapenem inactivation method (mCIM) and EDTA-CIM (eCIM) tests. MBL genes, phylogenetic group, and virulence genes profile of carbapenem resistant isolates were determined. Conjugation assay and plasmid profiling were conducted to evaluate the ability of transferring of CR to other E. coli isolates. Clonal similarity of isolates were assessed using Enterobacterial intergenic repetitive element sequence (ERIC)-PCR. RESULTS: Among 406 UPEC isolates, 12 (2.95%) carbapenem-resistant were detected of which 11 were phenotypically MBL-producing strains. Four isolates were resistant to all investigated antimicrobial agents and were considered possible pandrug-resistant (PDR). blaNDM, blaOXA-48, blaIMP-1, and blaIMP-2 genes were found in 9, 5, 1, and 1 isolates, respectively. Among 30 virulence genes investigated, the traT, fyuA followed by fimH, and iutA with the frequency of 8 (66.7%), 8 (66.7%), 7 (58.3%), and 7 (58.3%) were the most identified genes, respectively. Siderophore production was the main virulence trait among carbapenem-resistant UPEC isolates. Except for two, all other isolates showed weak to moderate virulence index. In all recovered isolates, CR was readily transmitted via plasmids to other isolates during conjugation experiments. CONCLUSION: MBL and carbapenemase genes, especially blaNDM and blaOXA-48 are spreading rapidly among bacteria, which can be a threat to global public health. Therefore monitoring the emergence and dissemination of new AMR is necessary to continuously refine guidelines for empiric antimicrobial therapy. Understanding the mechanisms of resistance and virulence in this group of bacteria can play an effective role in providing new therapeutic methods.
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Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/aislamiento & purificación , Factores de Virulencia/genética , beta-Lactamasas/genética , beta-Lactamasas/metabolismo , Antibacterianos/farmacología , Proteínas Bacterianas , Carbapenémicos/farmacología , Estudios Transversales , Farmacorresistencia Bacteriana/genética , Enterobacteriaceae/genética , Genotipo , Humanos , Irán , Pruebas de Sensibilidad Microbiana , Fenotipo , Filogenia , Plásmidos , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/efectos de los fármacos , VirulenciaRESUMEN
BACKGROUND: Uropathogenic Escherichia coli (UPEC) are frequent pathogens worldwide, impacting on the morbidity and economic costs associated with antimicrobial treatment. OBJECTIVES: We report two novel mutations associated with polymyxin-B resistance in an UPEC isolate collected in 2019. METHODS: Isolate was submitted to antimicrobial susceptibility testing including broth microdilution for polymyxin B. Whole genome was sequenced and analyzed. RESULTS: Polymyxin-B total inhibition occurred at 16 mg/L (resistant). UPEC isolate was assigned to the phylogroup D, serotype O117:H4, and Sequence Type 69. mcr genes were not detected, but two novel mutations in the pmrA/basS (A80S) and pmrB/basR (D149N) genes were identified. CONCLUSIONS: The occurrence of non-mcr polymyxin resistance in E. coli from extraintestinal infections underscores the need of a continuous surveillance of this evolving pathogen.
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Proteínas Bacterianas/genética , Farmacorresistencia Bacteriana Múltiple/genética , Factores de Transcripción/genética , Escherichia coli Uropatógena/genética , Antibacterianos/farmacología , Humanos , Mutación , Filogenia , Polimixina B/farmacología , Infecciones Urinarias/microbiología , Infecciones Urinarias/patología , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/aislamiento & purificaciónRESUMEN
Non-lactose-fermenting Escherichia coli (NLFEC) has a few descriptive studies restricted to human infections. In the present study, isolates of NLFEC obtained from urine samples of dogs with hyperadrenocorticism were characterized regarding their virulence ability, biofilm formation capacity and antimicrobial susceptibility profile. Escherichia coli lactose-fermenting strains from urinary infection in dogs with the same conditions were analysed to provide comparisons. The non-lactose-fermenting E. coli strains were classified as belonging to clade I E. coli, whereas the lactose-fermenting strains were classified in phylogroup B2. All strains presented virulence markers to adhesion, iron acquisition, toxins, colicin and cytotoxin production, and biofilm regulation. Components of the extracellular matrix in addition to the in vitro biofilm formation ability were observed in the strains. Multidrug resistance (MDR) profiles were observed by in vitro susceptibility tests to all NLFEC strains. In summary, non-lactose-fermenting uropathogenic E. coli from dogs behaves similar to lactose-fermenting E. coli, exhibiting MDR profile, and pathogenic potential of promote animal infections.
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Enfermedades de los Perros/microbiología , Infecciones por Escherichia coli/veterinaria , Infecciones Urinarias/veterinaria , Escherichia coli Uropatógena/patogenicidad , Factores de Virulencia/genética , Animales , Biopelículas/crecimiento & desarrollo , Perros , Farmacorresistencia Bacteriana Múltiple/genética , Fermentación/genética , Humanos , Filogenia , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/aislamiento & purificación , Escherichia coli Uropatógena/metabolismo , VirulenciaRESUMEN
BACKGROUND: Antibiotic resistance is an increasing phenomenon in many bacterial pathogens including uropathogenic Escherichia coli. Hypothetical anti-virulent agents could be a solution, but first clear virulence associated gene-pool of antibiotic resistant isolates have to be determined. The aim of this study is to investigate the significant associations between genes encoding VFs with antibiotic resistance and phylogenetic groups in UPEC isolates. RESULTS: The majority of 248 UPEC isolates belonged to phylogenetic group B2 (67.3%). The maximum and minimum resistance was attributed to amoxicillin (90.3%) and both fosfomycin and imipenem (1.6%) respectively. 11.3% of isolates were resistant to all antibiotic agents except that of imipenem, nitrofurantoin and fosfomycin. These highly resistant isolates were placed only in group B2 and D. The most prevalent virulence gene was ompA (93.5%). The hlyA was the only virulence gene that was significantly more prevalent in the highly resistant isolates. The ompA, malX and hlyA genes were obviously more abundant in the antibiotic resistant isolates in comparison to susceptible isolates. The papC gene was associated with amoxicillin resistance (p-value = 0.006, odds ratio: 26.00). CONCLUSIONS: Increased resistance to first line drugs prescribed for UTIs were detected in CA-UPEC isolates in our study.. Minimal resistance was observed against nitrofurantoin, fosfomycin and imipenem. Therefore, they are introduced for application in empirical therapy of UTIs. Fosfomycin may be the most effective antibiotic agent against highly resistant UPEC isolates. The presence of the ompA, malX and hlyA genes were significantly associated with resistance to different antibiotic agents. We assume that the ability of UPEC isolates to upgrade their antibiotic resistance capacity may occurs in compliance with the preliminary existence of specific virulence associated genes. But, more investigation with higher number of bacterial isolates, further virulence associated genes and comparison of gene pools from CA-UPEC isolates with HA-UPEC are proposed to confirm these finding and discovering new aspects of this association.
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Farmacorresistencia Bacteriana/genética , Filogenia , Escherichia coli Uropatógena/aislamiento & purificación , Factores de Virulencia/genética , Antibacterianos/farmacología , Infecciones Comunitarias Adquiridas/microbiología , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/genética , Humanos , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/genética , Virulencia/genéticaRESUMEN
Urinary tract infection (UTI) is caused by the invasion of the pathogen in the urinary system that can manifest as symptomatic or asymptomatic bacteriuria. This study was conducted to investigate antibiotic resistance patterns, and the correlation between biofilm formations with virulence factors in uropathogenic E. coli isolates retrieved from UTI. We searched Scopus and Google Scholar, PubMed, Web of sciences for studies published in the English language between 1st 2005 to 31st December 2019. The Mesh terms and text words included "biofilms", OR "biofilm formation", AND "antibiotic resistance", OR "drug-resistance", OR "antimicrobial drug resistance", AND "urinary tract infections", OR "UTI", AND "biofilm related-genes", AND "virulence factors" AND "correlation", AND "Uropathogenic Escherichia coli", OR "Uropathogenic E. coli" AND "prevalence" AND "Iran". Data analyzed using Comprehensive Meta-Analysis (CMA) software. The random-effects model was used to calculate the pooled prevalence with 95% confidence interval (CI). The combined rates of biofilm formation in Uropathogenic E. coli (UPEC) isolates were achieved as 84.6% (95% CI: 72.7-91.9). Also, 24.8%, 26.1% and 44.6% of UPEC isolates were able to create strong, moderate and weak biofilm, respectively. The highest pooled antibiotic resistance was against Ampicillin followed by Tetracycline with resistance rates of 74.6% and 64.9%, respectively. Accordingly, some studies reported that biofilm production was significantly associated with antibiotic resistance and virulence genes (p < 0.05). This study showed a high tendency among UPEC isolates to form biofilm (more than 84%), also, most studies included in the present review reported a significant correlation between biofilm formation with antibiotic resistance and virulence factors.
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Antibacterianos/farmacología , Biopelículas/crecimiento & desarrollo , Farmacorresistencia Bacteriana/efectos de los fármacos , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/aislamiento & purificación , Biopelículas/efectos de los fármacos , Bases de Datos Factuales , Farmacorresistencia Bacteriana/genética , Infecciones por Escherichia coli/epidemiología , Humanos , Irán/epidemiología , Pruebas de Sensibilidad Microbiana , Infecciones Urinarias/tratamiento farmacológico , Escherichia coli Uropatógena/genética , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
The unique plasmonic energy exchange occurring within metallic crossed surface relief gratings (CSRGs) has recently motivated their use as biosensors. However, CSRG-based biosensing has been limited to spectroscopic techniques, failing to harness their potential for integration with ubiquitous portable electronics. Here, we introduce biosensing via surface plasmon resonance imaging (SPRi) enabled by CSRGs. The SPRi platform is fully integrated including optics and electronics, has bulk sensitivity of 613 Pixel Intensity Unit (PIU)/Refractive Index Unit (RIU), a resolution of 10-6 RIU and a signal-to-noise ratio of â¼33 dB. Finite-Difference Time-Domain (FDTD) simulations confirm that CSRG-enabled SPRi is supported by an electric field intensity enhancement of â¼30 times, due to plasmon resonance at the metal-dielectric interface. In the context of real-world biosensing applications, we demonstrate the rapid (<35 min) and label-free detection of uropathogenic E. coli (UPEC) in PBS and human urine samples for concentrations ranging from 103 to 109 CFU mL-1. The detection limit of the platform is â¼100 CFU mL-1, three orders of magnitude lower than the clinical detection limit for diagnosis of urinary tract infection. This work presents a new avenue for CSRGs as SPRi-based biosensing platforms and their great potential for integration with portable electronics for applications requiring in situ detection.
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Infecciones por Escherichia coli/orina , Resonancia por Plasmón de Superficie/instrumentación , Escherichia coli Uropatógena/aislamiento & purificación , Diseño de Equipo , Humanos , Límite de Detección , Nanoestructuras/química , Nanoestructuras/ultraestructura , Refractometría/instrumentación , Refractometría/métodos , Resonancia por Plasmón de Superficie/métodos , Propiedades de SuperficieRESUMEN
BACKGROUND: Urinary tract infection (UTI) is a common cause of morbidity worldwide. Uropathogenic Escherichia coli (UPEC) bacteria are the major cause of urinary tract infections. UPEC strains derive from different phylogenetic groups and possess an arsenal of virulence factors that contribute to their ability to overcome different defense mechanisms and cause disease. The objective of this study was to identify phylogroup and virulence genes of UPEC among urinary tract infection patients. METHODS: A cross sectional study was conducted from January 1, 2017 to October 9, 2017. E. coli bacteria were isolated from UTI patients using culture and conventional biochemical tests. Identification of phylogroup and genes that encodes for virulence factors was done using multiplex polymerase chain reaction (PCR). Data was processed and analyzed with SPSS version16.0 and Epi-info version 3.4.1 software. RESULTS: The most common urologic clinical manifestation combinations in this study were dysuria, urine urgency and urgency incontinence. The frequent UPEC virulence gene identified was fimH 164 (82%), followed by aer 109 (54.5%), hly 103 (51.5%), pap 59 (29.5%), cnf 58 (29%), sfa 50 (25%) and afa 24 (12%).There was significant association between pap gene and urine urgency (p-0.016); sfa and dysuria and urine urgency (p-0.019 and p-0.043 respectively); hly and suprapubic pain (p-0.002); aer and suprapubic pain, flank pain and fever (p-0.017, p-0.040, p-0.029 respectively). Majority of E. coli isolates were phylogroup B2 60(30%) followed by D 55(27.5%), B1 48(24%) and A 37(18.5%). There was significant association between E. coli phylogroup B2 and three virulence genes namely afa, pap, and sfa (p-0.014, p-0.002, p-0.004 respectively). CONCLUSION: In this study the most frequent E. coli virulence gene was fimH, followed by aer, hly, pap, cnf, sfa and afa respectively. There was significant association between E. coli virulence genes and clinical symptoms of UTI. The phylogenetic analysis indicates majority of uropathogenic E. coli isolates were phylogroup B2 followed by phylogroup D. Phylogroup B2 carries more virulence genes. Hence, targeting major UPEC phylogroup and virulence genes for potential vaccine candidates is essential for better management of UTI and further research has to be conducted in this area.
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Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/genética , Genes Bacterianos , Filogenia , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/patogenicidad , Virulencia/genética , Adulto , Estudios Transversales , Etiopía , Femenino , Humanos , Masculino , Persona de Mediana Edad , Encuestas y Cuestionarios , Escherichia coli Uropatógena/aislamiento & purificación , Factores de Virulencia/genéticaRESUMEN
INTRODUCTION: Uropathogenic E. coli is the leading cause of Urinary tract infections (UTIs), contributing to 80-90% of all community-acquired and 30-50% of all hospital-acquired UTIs. Biofilm forming Uropathogenic E. coli are associated with persistent and chronic inflammation leading to complicated and or recurrent UTIs. Biofilms provide an environment for poor antibiotic penetration and horizontal transfer of virulence genes which favors the development of Multidrug-resistant organisms (MDRO). Understanding biofilm formation and antimicrobial resistance determinants of Uropathogenic E. coli strains will provide insight into the development of treatment options for biofilm-associated UTIs. The aim of this study was to determine the biofilm forming capability, presence of virulence genes and antimicrobial susceptibility pattern of Uropathogenic E. coli isolates in Uganda. METHODS: This was a cross-sectional study carried in the Clinical Microbiology and Molecular biology laboratories at the Department of Medical Microbiology, Makerere University College of Health Sciences. We randomly selected 200 Uropathogenic E. coli clinical isolates among the stored isolates collected between January 2018 and December 2018 that had significant bacteriuria (> 105 CFU). All isolates were subjected to biofilm detection using the Congo Red Agar method and Antimicrobial susceptibility testing was performed using the Kirby disk diffusion method. The isolates were later subjected PCR for the detection of Urovirulence genes namely; Pap, Fim, Sfa, Afa, Hly and Cnf, using commercially designed primers. RESULTS: In this study, 62.5% (125/200) were positive biofilm formers and 78% (156/200) of these were multi-drug resistant (MDR). The isolates were most resistant to Trimethoprim sulphamethoxazole and Amoxicillin (93%) followed by gentamycin (87%) and the least was imipenem (0.5%). Fim was the most prevalent Urovirulence gene (53.5%) followed by Pap (21%), Sfa (13%), Afa (8%), Cnf (5.5%) and Hyl (0%). CONCLUSIONS: We demonstrate a high prevalence of biofilm-forming Uropathogenic E. coli strains that are highly associated with the MDR phenotype. We recommend routine surveillance of antimicrobial resistance and biofilm formation to understand the antibiotics suitable in the management of biofilm-associated UTIs.
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Antibacterianos/uso terapéutico , Biopelículas/crecimiento & desarrollo , Infecciones por Escherichia coli/epidemiología , Infecciones Urinarias/epidemiología , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/patogenicidad , Estudios Transversales , Farmacorresistencia Bacteriana Múltiple/genética , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/microbiología , Humanos , Pruebas de Sensibilidad Microbiana , Fenotipo , Reacción en Cadena de la Polimerasa , Prevalencia , Uganda/epidemiología , Infecciones Urinarias/tratamiento farmacológico , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/aislamiento & purificación , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
BACKGROUND: Disease prevention and control is a significant part in the ex-situ conservation of the endangered red panda (Ailurus fulgens), being bacterial infection is one of the most important health threats to the captive population. To date, studies about the infection caused by Escherichia coli in the red panda are scarce. This study was conducted to determine the cause of death of a captive red panda through clinical symptoms, complete blood count, biochemical analysis, pathological diagnosis and bacterial whole genome sequencing. CASE PRESENTATION: The following report describes a case of a 1.5 year old captive red panda (Ailurus fulgens) that was found lethargic and anorectic. She was moved to the quarantine area for daily treatment with 50 mg of Cefpodoxime Proxetil. During the three-day treatment, she did not eat or defecate, and then died. Clinical hematology revealed the values of neutrophils, alanine aminotransferase (ALT), aspartate aminotransferase (AST) and blood urea nitrogen (BUN) were significantly higher. Histological analysis demonstrated major pathological damage in the kidneys, liver and lungs, characterized by hyperemia, parenchymal cell degeneration and necrosis and inflammatory cell infiltration which were predominantly neutrophilic. A bacterial strain confirmed as Escherichia coli was isolated post mortem. Whole genome sequencing of the E. coli showed the complete genome size was 4.99 Mbp. PapA, PapC, OmpA, OmpU and other virulence factors which specific to Uropathogenic Escherichia coli (UPEC) were found in the isolate. Among the virulence factors, P pili, type I pili and related factors of the iron uptake system were associated with nephrotoxicity. CONCLUSION: The red panda died of bacterial infection caused by an uropathogenic strain of Escherichia coli. The pathogenic mechanisms of the strain are closely related to the expression of specific virulence genes.
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Ailuridae , Infecciones por Escherichia coli/veterinaria , Escherichia coli Uropatógena/aislamiento & purificación , Animales , Antibacterianos/uso terapéutico , Ceftizoxima/análogos & derivados , Ceftizoxima/uso terapéutico , Infecciones por Escherichia coli/tratamiento farmacológico , Infecciones por Escherichia coli/microbiología , Femenino , Genoma Bacteriano , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/patogenicidad , Factores de Virulencia/genética , Secuenciación Completa del Genoma/veterinaria , Cefpodoxima ProxetiloRESUMEN
The spread of multidrug resistant bacteria has become a global concern. One of the most important and emergent classes of multidrug-resistant bacteria is extended-spectrum ß-lactamase-producing bacteria (ESBL-positive = ESBL+). Due to widespread and continuous evolution of ESBL-producing bacteria, they become increasingly resistant to many of the commonly used antibiotics, leading to an increase in the mortality associated with resulting infections. Timely detection of ESBL-producing bacteria and rapid determination of their susceptibility to appropriate antibiotics can reduce the spread of these bacteria and the consequent complications. Routine methods used for the detection of ESBL-producing bacteria are time-consuming, requiring at least 48 h to obtain results. In this study, we evaluated the potential of infrared spectroscopic microscopy, combined with multivariate analysis for rapid detection of ESBL-producing Escherichia coli ( E. coli) isolated from urinary-tract infection (UTI) samples. Our measurements were conducted on 837 samples of uropathogenic E. coli (UPEC), including 268 ESBL+ and 569 ESBL-negative (ESBL-) samples. All samples were obtained from bacterial colonies after 24 h culture (first culture) from midstream patients' urine. Our results revealed that it is possible to detect ESBL-producing bacteria, with a 97% success rate, 99% sensitivity, and 94% specificity for the tested samples, in a time span of few minutes following the first culture.
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Rayos Infrarrojos , Aprendizaje Automático , Microscopía , Escherichia coli Uropatógena/aislamiento & purificación , Escherichia coli Uropatógena/metabolismo , beta-Lactamasas/biosíntesis , Espectroscopía Infrarroja por Transformada de FourierRESUMEN
BACKGROUND: Escherichia coli has found to be the predominant uropathogen (50-90%) in uncomplicated, community acquired urinary tract infection (UTI). Uropathogenic Escherichia coli (UPEC) express a multitude of virulence factors, which enable the bacteria to establish UTI. The objective of this study was to evaluate the presence of different phenotypic virulence markers in UPEC isolates and determine their correlation with antibiotic resistance pattern. RESULTS: Out of 105 patients, 56 (53%) were females and 49 (47%) were males. The age of the patients in the study ranged from 18 years to 87 years and majority of the patients belonged to the age group 20-29 years. Virulence factor was observed in 65% (n = 69) of UPEC and 20% (n = 22) of control isolates (P = 0.0001). Haemolysin production was observed in 34(32.3%) of uroisolates and 12 (11.4%) of control strain. Similarly, 62% of UPEC and 1% of control produced biofilm (P = 0.0001). The expression of Mannose-resistant hemagglutinin (MRHA) and mannose-sensitive hemagglutinin (MSHA) in uroisolates were 52.3% (n = 55) and 5.7% (n = 6) respectively, whereas in faecal isolates, 8.5% (n = 9) expressed MRHA and none produced MSHA. Antimicrobial resistance showed a high degree of resistance towards ampicillin, cotrimoxazole and norfloxacin. The resistance was observed in significant higher degree in biofilm formers as compared to non-formers. MDR and ESBL was observed in 51 and 46% of test strains and 9 and 7.6% of control strains (P = 0.0001). CONCLUSION: A significant association between virulence factors of UPEC and antimicrobial resistance in UPEC was present. Routine testing of these factors and co-relation with AMR is recommended. These findings will certainly help understand the pathogenicity and proper management of UTI patients, thus decreasing the improper use of antibiotics.
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Farmacorresistencia Bacteriana , Infecciones por Escherichia coli/microbiología , Proteínas de Escherichia coli/genética , Escherichia coli Uropatógena/efectos de los fármacos , Factores de Virulencia/genética , Adolescente , Adulto , Anciano , Anciano de 80 o más Años , Antibacterianos/farmacología , Proteínas de Escherichia coli/metabolismo , Femenino , Humanos , Masculino , Pruebas de Sensibilidad Microbiana , Persona de Mediana Edad , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/genética , Escherichia coli Uropatógena/aislamiento & purificación , Factores de Virulencia/metabolismo , Adulto JovenRESUMEN
To characterise the dissemination patterns of uropathogenic Escherichia coli (UPEC) in a community, we conducted a study utilising molecular and fundamental descriptive epidemiology. The subjects, consisted of women having community-acquired acute urinary tract infection (UTI), were enrolled in the study from 2011 to 2012. UPEC isolates were subjected to antibacterial-susceptibility testing, O serogrouping, phylotyping, multilocus-sequence typing with phylogenetic-tree analysis and pulsed-field-gel electrophoresis (PFGE). From the 209 unique positive urinary samples 166 UPEC were isolated, of which 129 were fully susceptible to the tested antibiotics. Of the 53 sequence types (STs), the four most prevalent STs (ST95, ST131, ST73 and ST357) accounted for 60% of all UPEC strains. Antimicrobial resistance was less frequently observed for ST95 and ST73 than for the others. A majority of rare STs and a few common STs constituted the diversity pattern within the population structure, which was composed of the two phylogenetically distinct clades. Eleven genetically closely related groups were determined by PFGE, which accounted for 42 of the 166 UPEC isolates, without overt geo-temporal clustering. Our results indicate that a few major lineages of UPEC, selected by unidentified factors, are disseminated in this community and contribute to a large fraction of acute UTIs.
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Infecciones Comunitarias Adquiridas/epidemiología , Infecciones por Escherichia coli/epidemiología , Genotipo , Infecciones Urinarias/epidemiología , Escherichia coli Uropatógena/aislamiento & purificación , Infecciones Comunitarias Adquiridas/microbiología , Infecciones Comunitarias Adquiridas/transmisión , Infecciones por Escherichia coli/microbiología , Infecciones por Escherichia coli/transmisión , Femenino , Humanos , Pruebas de Sensibilidad Microbiana , Epidemiología Molecular , Tipificación Molecular , Serotipificación , Infecciones Urinarias/microbiología , Infecciones Urinarias/transmisión , Escherichia coli Uropatógena/clasificación , Escherichia coli Uropatógena/efectos de los fármacos , Escherichia coli Uropatógena/genéticaRESUMEN
BACKGROUND: Escherichia coli lineage ST131 predominates across various spectra of extra-intestinal infections, including urinary tract infection (UTI). The distinctive resistance profile, diverse armamentarium of virulence factors and rapid global dissemination of ST131 E. coli makes it an intriguing pathogen. However, not much is known about the prevalence and genetic attributes of ST131 lineage in Pakistan. METHODS: We estimated prevalence and genetic attributes of E. coli ST131 isolates causing UTI among 155 randomly selected samples. Samples were analyzed for phylogenetic grouping, O-typing and fumC/fimH typing. Isolates were further tested for the ESBL and virulence factors using PCR. RESULTS: Overall, 59% of the UPEC isolates belonged to the phylogenetic group B2, followed by D = 28%, B1 = 8% and A = 5%. Among 18 different Sequence-types, ST131 was the dominant lineage (n = 71; 46%) out of which 72% of the isolates were assigned to the phylogenetic group B2, while 61% adhered to the serogroup O25b. FumC/fimH typing confirmed 49% of the ST131 as H30 sub-types. In this study, significant numbers of the identified ST131 isolates were MDR and 42% showed ESBL phenotypes, out of which 37% carried bla-CTX-M-15. Moreover, different virulence factors were detected in following percentages: fimH,155(100%), iutA 86 (55%), feoB 76 (49%), papC 75 (48%), papGII 70 (45%), kpsMTII 40 (26%), papEF 37 (24%), fyuA 37 (24%), usp 22 (14%), papA 20 (13%), sfa/foc20 (13%), hlyA 18 (12%), afa 15 (10%), cdtB 11 (7%), papGI 6 (4%), papGIII 6 (4%), kpsMTIII 4 (3%) and bmaE2 (1%). CONCLUSION: Conclusively, this study provides important insight into the genetic and virulence attributes of pandemic MDR ST131 strains involved in UTIs. It also highlights higher prevalence of ST131-O25b-H30 UPEC isolates in patients, which was previously unreported from this part of globe.
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Infecciones por Escherichia coli/microbiología , Infecciones Urinarias/diagnóstico , Escherichia coli Uropatógena/clasificación , Farmacorresistencia Bacteriana Múltiple/genética , Infecciones por Escherichia coli/diagnóstico , Infecciones por Escherichia coli/epidemiología , Femenino , Humanos , Masculino , Pakistán/epidemiología , Filogenia , Prevalencia , Infecciones Urinarias/epidemiología , Infecciones Urinarias/microbiología , Escherichia coli Uropatógena/aislamiento & purificación , Escherichia coli Uropatógena/patogenicidad , Virulencia , Factores de Virulencia/clasificación , Factores de Virulencia/genética , beta-Lactamasas/clasificación , beta-Lactamasas/genéticaRESUMEN
With the multiplicity of existing methods to track E. coli infections, it still seems necessary to seek new, better and/or complementary ways for epidemiological investigations. Particularly, fast, cheap, effective and reproducible methods providing easily comparable results are needed. Our previous studies showed that the use of TRS-PCR is an effective molecular tool in E. coli epidemiology. In this paper, we have developed a unique classification scheme in which an individual TRS-PCR pattern is assigned a numerical value. This approach allows for rapid interpretation of the results obtained from several similarity dendrograms. Using this approach, based on CAC-PCR, GTG-PCR and CGG-PCR, we obtained 52, 86 and 99 different numerical types for the 124 analyzed uropathogenic E. coli strains, respectively. This allowed for the identification of 121 unique isolates differing in at least one TRS-PCR class. In this approach, we got numerical results, easy to sort and interpret, allowing easier analysis of these strains.
Asunto(s)
Bacteriuria/genética , Escherichia coli/genética , Perfilación de la Expresión Génica/métodos , Bacteriuria/microbiología , Escherichia coli/clasificación , Infecciones por Escherichia coli , Femenino , Perfil Genético , Humanos , Masculino , Polonia/epidemiología , Reacción en Cadena de la Polimerasa/métodos , Repeticiones de Trinucleótidos/genética , Escherichia coli Uropatógena/aislamiento & purificaciónRESUMEN
AIMS: The aim of this study was to assess anti-biofilm and antimicrobial effects of auranofin, an anti-rheumatic agent, on uropathogenic Escherichia coli (UPEC) biofilm formation. METHODS AND RESULTS: The minimum inhibitory concentration and biofilm inhibition concentration of auranofin against UPEC ranged from 24 to 32 µg ml-1 . Biofilm eradication concentration and XTT reduction concentration of auranofin were found to be at 112 µg ml-1 . Confocal laser scanning microscopy results confirmed that biofilm was inhibited by auranofin. These results indicate that auranofin possesses potent anti-biofilm and antimicrobial activities against UPEC. Effects of auranofin on type 1 fimbriae gene (fimH) and response regulator gene (rpoS) to stress were explored using quantitative real time-polymerase chain reaction. In addition, combination of auranofin and tetracycline showed synergistic effect. CONCLUSIONS: These data indicate that auranofin has inhibitory effect on biofilm formation and synergistic effect on UPEC infection when it is combined with tetracycline. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study strongly suggest that auranofin is a promising alternative anti-biofilm and antimicrobial agent to prevent UPEC biofilm formation in UTIs. Auranofin already approved for human use have the advantage of being able to be put into clinical use relatively quickly.