Clinico-microbiological Profile of Urinary Tract Infection with Special Reference to Uropathogenic E coli : Antibiotic susceptibility Pattern, Phylogenetic Background and Virulent Factor Distribution from West Bengal, India

Background and Objectives : Routine surveillance and monitoring studies pose a constant need to update clinicians on prevalent pathogens and rational and empirical treatment in Urinary Tract Infection (UTI). Escherichia coli (E coli) is the most commonly isolated uropathogen globally. Extended-Spectrum ?-Lactamase (ESBL) production and ?-Lactamase Inhibitor Resistance (BLIR) among these pathogens together with their uro-virulence determinants further complicate treatment approaches. This study investigated the clinico-microbiological pattern of UTI and determined the antibiotic sensitivity pattern, the phylogenetic background, and virulence determinants of E coli, the most commonly isolated uropathogen. Methods : Uropathogens isolated by urine culture from community and hospitalized patients were biochemically speciated. Antibiotic susceptibility was tested by Kirby-bauer disk diffusion method. Phylogenetic background and virulence determinants of E coli isolates were identified by PCR. SPSS 16.0 was used for statistical interpretation. Results : 45% of the urine samples showed growth positivity. 44% amongst them were E coli. All isolates were multidrug-resistant. 50% and 40% were ESBL producers and BLIR respectively. Former showed highest resistance to quinolone, fluoroquinolones, cotrimoxazole, and latter were resistant against all drugs tested except nitrofurantoin. Significant correlation existed between the ?-lactams, quinolone, fluoroquinolones, cotrimoxazole (p<0.05) resistance pattern. BLIR and ESBL E coli recorded highest prevalence of pathogenic phylogroup B2 and D respectively. Varied prevalence of fimbrial (fimH, papC, papEF, papG, GII) and toxin genes (iroN, hlyA, cnfI, i ucD, cdtBU) in ESBL, BLIR and non-ESBL isolates were observed. Their distribution was statistically significant (p=0.05). Interpretation and Conclusions : Nitrofurantoin is the drug of choice in empirical treatment of uncomplicated UTI. Aggressive and consistent investigation and health education are highly recommended for effective clinical management in UTI.

ß-lactamasas de espectro extendido y factores de virulencia en Escherichia coli uropatógenas en asilos de ancianos en Lima, Perú / ß-lactamases and virulence factors in uropathogenic Escherichia coli in nursing homes in Lima, Peru

RESUMEN Los asilos de ancianos son instituciones con una alta prevalencia de infecciones del tracto urinario ocasionado por Escherichia coli productoras de ß-lactamasas de espectro extendido (BLEE), con diversos factores de virulencia. El objetivo del estudio fue determinar la frecuencia del gen bla CTX-M y de ocho genes de virulencia en 35 E. coli uropatógenas productoras de BLEE provenientes de seis asilos en Perú, durante el 2018. El 57,1% (20/35) de las E. coli fueron portadores del gen bla CTX-M. Además, se obtuvo una frecuencia del 46% (15/35) y 37% (13/35) de hly-alfa y cnf-1, respectivamente; elevada presencia de los genes iucC (63%, 22/35), aer (94%, 33/35) y chuA (94%, 33/34) y una frecuencia del 46% (16/35) y del 91% (32/34) de los genes pap GII y nanA, respectivamente. Existe predominancia en la distribución del gen bla CTX-M, además de una alta frecuencia de exotoxinas que le confieren una ventaja competitiva para diseminarse hacia el torrente sanguíneo.

ABSTRACT Nursing homes are institutions with high prevalence of urinary tract infections caused by ESBL-producing E. coli with several virulence factors. The aim of this study was to determine the frequency of the bla CTX-M gene and eight virulence genes in 35 ESBL-producing uropathogenic E. coli from six nursing homes in Peru during 2018. Of the E. coli samples, 57.1% (20/35) were carriers of the bla CTX-M gene. Furthermore, we obtained frequencies of 46% (15/35) and 37% (13/35) for hly-alpha and cnf-1, respectively; we also found high presence of the iucC (63%, 22/35), aer (94%, 33/35) and chuA genes (94%, 33/34) as well as a frequency of 46% (16/35) and 91% (32/34) for the pap GII and nanA genes, respectively. The bla CTX-M gene is predominant and a high frequency of exotoxins gives it a competitive advantage for spreading into the bloodstream.

Role and pathogenic mechanism of TcpC in uropathogenic Escherichia coli-induced mouse model of cystitis / 中华微生物学和免疫学杂志

Objective:To investigate the role of TcpC in uropathogenic Escherichia coli (UPEC)-induced cystitis in mice and to preliminarily analyze the pathogenic mechanism. Methods:C57BL/6 mice were injected with 10 9 CFU wild-type UPEC CFT073 (CFT073 wt) or tcpc gene-deleted mutant (CFT073 Δ tcpc) from urethra into bladder to construct the mouse model of cystitis. The mice were sacrificed 3 d after infection and the bladders were taken to observe the gross pathological changes. Histopathological changes in bladder tissues were observed after HE staining. Immunohistochemistry was used to detect TcpC in bladder tissues. Bacterial loads in urine samples of UPEC-infected mice were counted by tenfold dilution method, and the presence of tcpc gene in the genomic DNA of bacteria from the bladder and urine samples of CFT073 wt-infected mice was measured by PCR. Real-time quantitative RT-PCR (qRT-PCR) and Western blot were performed to detect the expression of TcpC at mRNA and protein levels in macrophages after CFT073 wt infection. The influence of UPEC strains on the activation of NF-κB signaling pathway in macrophages were determined by Western blot. The levels of proinflammatory factors and the bacterial and cell activity after infecting macrophages with UPEC strains were detected by ELISA, laser confocal microscope and fluorescence microscope, respectively. Results:Compared with the mice with CFT073 Δ tcpc infection, CFT073 wt-infected mice had significantly enlarged bladder and severe neutrophil infiltration and abundant TcpC in bladder tissues. The number of bacteria in the urine of CFT073 wt-infected mice was significantly greater than that of the CFT073 Δ tcpc group. PCR results showed that the bacteria in bladder or urine were CFT073 wt. The expression of TcpC at both mRNA and protein levels in macrophages increased significantly after CFT073 wt infection. Moreover, in CFT073 wt-infected macrophages, the expression of IκBα was promoted and the phosphorylation of p65 and the production of proinflammatory factors were suppressed. TcpC was instrumental in the survival and invasion of CFT073 wt in macrophages. Conclusions:TcpC expression increased significantly in mice with CFT073 wt-induced cystitis. TcpC inhibited the activation of NF-κB signaling pathway and the production of proinflammatory factors in macrophages to improve the survival rate of CFT073 wt, which was closely related to the pathogenesis and immune evasion of UPEC.

Signaling pathways involved in inhibition of macrophage phagocytosis by TcpC of uropathogenic Escherichia coli / 中华微生物学和免疫学杂志

Objective:To investigate the signaling pathway of inhibiting macrophage phagocytosis of TIR domain-containing protein encoded by Escherichia coli (TcpC) N-terminal ubiquitin ligase active fragments of uropathogenic Escherichia coli. Methods:Bioinformatics software was used to analyze the amino acid sequences and the function of TcpC N-terminal ubiquitin ligase active fragments as well as the functional sites. PCR was performed to amplify tcpc-330, tcpc-450 and tcpc-510 genes and a prokaryotic expression system was constructed to express the target proteins. The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were purified by Ni-NTA affinity chromatography. LPS in the recombinant proteins was removed by Detoxi-gel chromatography. The expression of MyD88 at protein and mRNA levels in macrophages incubated with rTcpC-N110, rTcpC-N150, rTcpC-N170 or rTcpC-TIR was detected by Western blot and qRT-PCR. The activation of NF-κB signal pathway and the levels of proinflammatory factors in macrophages incubated with the above TcpC protein fragments were measured by Western blot and ELISA, respectively. Results:Cys12, Trp104 and Trp106 in the N-terminal fragment of TcpC were crucial amino acids in maintaining its ubiquitin ligase activity. The target recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 were successfully expressed and purified. After Detoxi-gel chromatography, rTcpC-N110, rTcpC-N150 and rTcpC-N170 extracts were undetectable for LPS. TcpC ubiquitin ligase fragments inhibited the expression of MyD88 at protein level, but not affect its expression at mRNA level in macrophages. LPS-induced phosphorylation of NF-κB signaling pathway-related proteins p50 and p65 was significantly inhibited in macrophages treated with TcpC ubiquitin ligase fragments. Moreover, LPS-induced production of pro-inflammatory factors was also significantly inhibited.Conclusions:The recombinant proteins rTcpC-N110, rTcpC-N150 and rTcpC-N170 could inhibit the expression of MyD88 at protein level and suppress the activation of NF-κB signaling pathway, suggesting that they were closely related to the inhibition of innate immune activity of macrophages.

Susceptibility to first choice antimicrobial treatment for urinary tract infections to Escherichia coli isolates from women urine samples in community South Brazil

ABSTRACT E. coli is the main pathogen of UTI. It is important to be aware the local epidemiological data for an appropriate initial treatment. Resistance to antimicrobial agents has increased, especially to first-choice antibiotics in the treatment of cystitis. There are few studies on the sensivity profile of community uropathogen in our region. Objective: To characterize antimicrobials the sensitivity profile to E. coli isolated from urocultures of women treated at Basic Health Units and Emergency Care Units of Londrina-Paraná- Brazil during a period of 12 months (June 1, 2016 to June 1, 2017). Methodology: A cross-sectional study was carried out from June 2016 to June 2017. All urine samples collected in the Basic Health Units and Emergency Departments in the city of Londrina (Paraná State, Brazil) were sent to a Central Laboratory where the identification and antimicrobial susceptibility testing were performed. Clinical Laboratory Standards Institute (CLSI) breakpoints were used for the interpretation of susceptibility testing results. Results: 56,555 urine cultures were performed in the period, of which 8,832 were positive, of which 5,377 were women. Of these samples, 4.7% were enterobacteria producing extended-spectrum beta-lactamases (ESBL) and 15.5% resistant to quinolones. TMP- SMX was resistant in more than 30% of the samples in all age groups. Among quinolone-resistant isolates, resistance to cephalothin, ampicillin and sulfamethoxazole-trimethoprim was greater than 60%. Nitrofurantoin was the only antimicrobial that showed 90% of sensitivity. Conclusion: The antimicrobials sensitivity profile was similar to that reported in the literature, with TMP- SMX resistance greater than 30% in the studied samples. Nitrofurantoin maintains high sensitivity rates greater than 90%. Resistance to quinolones increases proportionally with age, as well ESBL.

Bilateral pyelonephritis due to Escherichia coli infection in a captive jaguar (Panthera onca) / Pielonefrite bilateral por Escherichia coli em uma onça-pintada (Panthera onca)

Extraintestinal pathogenic Escherichia coli (ExPEC) is a highly diverse pathotype of E. coli which colonizes the intestine, and it is considered an important etiological agent associated with bacteremia and other systemic infections, among them urinary tract infection. Retrospective studies evaluating morbidity and mortality of nondomestic felids have demonstrated that urinary tract diseases are among the main causes of death for geriatric animals. Also, mesenchymal neoplasms of the uterus are common in wild felids, and they possess variable morphologic characteristics related to invasiveness and malignancy. This report describes a case of bilateral pyelonephritis due to extraintestinal uropathogenic E. coli infection in a captive jaguar (Panthera onca). The diagnosis was confirmed through pathological, bacterial and immunohistochemical findings. According to molecular analysis, this E. coli strain was classified in the phylogroup F, possessing the following virulence-associated genes: usp, cnf-1, hlyA, papC and sfa. Additionally, this E. coli was highly resistant to beta-lactams and first-generation cephalosporin. This jaguar also presented a uterine leiomyoma with distinct distribution, and severe degenerative articular disease, both of them described as frequently seen lesions in geriatric animals from the Panthera genus.(AU)

Escherichia coli extraintestinal patogênica (ExPEC) é um patotipo altamente diverso de E. coli que coloniza o intestino e é considerada um agente etiológico importante, associado com bacteremia e outras infecções sistêmicas, dentre elas infecções do trato urinário. Estudos retrospectivos avaliando morbidade e mortalidade de felídeos não domésticos demostram que doenças do trato urinário estão entre as principais causas de morte de animais geriátricos. Ainda, neoplasias mesenquimais uterinas são comuns em felídeos de cativeiro e possuem características morfológicas variáveis relacionadas a invasividade e malignidade. Neste relato é descrito um caso de pielonefrite bilateral por E. coli extraintestinal uropatogênica em uma onça-pintada de cativeiro (Panthera onca). O diagnóstico foi confirmado através dos achados patológicos, bacteriológicos e imuno-histoquímicos. A partir da análise molecular, esta cepa de E. coli foi classificada no filogrupo F, possuindo os seguintes genes associados a virulência: usp, cnf-1, hlyA, papC and sfa. Adicionalmente, a bactéria isolada foi altamente resistente a ß-lactâmicos e cefalosporinas de primeira geração. Foi observado ainda um leiomioma uterino com distribuição distinta e doença articular degenerativa severa, ambas descritas na literatura como comumente observadas em animais geriátricos do gênero Panthera.(AU)

Respuesta inmunológica y bioquímica de ancianos con infección urinaria frente factores de virulencia en Escherichia coli uropatógenas / Immunological and biochemical response of older adults with urinary tract infection to uropathogenic Esherichia coli virulence factors

RESUMEN Con el objetivo de evidenciar diferencias en la respuesta inmunológica y bioquímica de ancianos con infección del tracto urinario (ITU) por Escherichia coli (E. coli) frente a factores de virulencia de importancia en la patogenia de la ITU, se realizó un estudio descriptivo en el cual se evaluaron 24 muestras de orina de adultos mayores con ITU provenientes de centros de reposo gerontológicos. Se determinó la concentración de hierro, TNF-α, IL-1β y la capacidad antioxidante en la orina, encontrándose una relación entre una mayor concentración de hierro y de hematíes en la orina con la presencia del gen pap GII en la E. coli. Se concluye que los adultos mayores con ITU por E. coli portadoras del gen pap GII, presentan mayor daño tisular.

ABSTRACT Descriptive study in which 24 urine samples from older adults with urinary tract infection (UTI), from nursing homes, were evaluated; in order to identify differences in the immune and biochemical response from older adults with UTI by Escherichia coli (E. coli) to major virulence factors in the pathogenesis of UTI. Iron concentration, TNF-α, IL-1β and antioxidant capacity in urine were determined. A relation was found between, an increase in iron and red blood cell concentration in urine, and the presence of the pap GII gene found in E. coli. It is concluded that older adults, with UTIs by E. coli with the gene pap GII, have increased tissue damage.

Presencia de genes fimH y afa en aislamientos urinarios de Escherichia coli productora de betalactamasas de espectro extendido en Lima, Perú / Presence of fimH and afa genes in urinary isolates of extended-spectrum beta-lactamases producing Escherichia coli in Lima, Peru

RESUMEN Con el objetivo de determinar la presencia de los genes fimH y afa en aislamientos urinarios de Escherichia coli productoras de betalactamasas de espectro extendido (BLEE), se realizó un estudio descriptivo, con aislamientos del cepario del proyecto TO-06/09 del Instituto Nacional de Salud del Niño en Lima, Perú. Se incluyeron 75 aislamientos urinarios de Escherichia coli. La identificación de genes se realizó por reacción en cadena de la polimerasa. De los 75 aislamientos, 74 (98,7%) fueron positivos para el gen fimH y 6 (8,0%) fueron positivos para el gen afa. Se evidenció la presencia de los factores de virulencia producidos por los genes fimH y afa en aislamientos urinarios de Escherichia coli productoras de BLEE.

ABSTRACT Descriptive study conducted in order to determine the presence of the fimH and afa genes in urinary isolates of extended-spectrum beta-lactamases (ESBL) producing Escherichia coli. Isolates from project TO-06/09 of the Instituto Nacional de Salud del Niño in Lima, Peru were used. A total of 75 urinary isolates of Escherichia coli were included. Gene identification was performed by polymerase chain reaction. From the 75 isolates, 74 (98.7%) were positive for the fimH gene and 6 (8.0%) were positive for the afa gene. Virulence factors produced by the fimH and afa genes were evident in urinary isolates of ESBL producing Escherichia coli.

Distribución de grupos filogenéticos, factores de virulencia y susceptibilidad antimicrobiana en cepas de Escherichia coli uropatógena / Distribution of phylogenetic groups, virulence factors and antimicrobial susceptibility in strains of uropathogenic Escherichia coli

Resumen Introducción: La diferencia entre los aislados patógenos y comensales de Escherichia coli se fundamenta en sus antecedentes filogenéticos. En Venezuela son escasos los estudios que describen el potencial patogénico de los grupos filogenéticos en E. coli. Objetivo: Relacionar la susceptibilidad antimicrobiana, distribución de los grupos filogenéticos y genes de virulencia en cepas de E. coli uropatógena (ECUP) aisladas de pacientes con infección del tracto urinario. Materiales y Métodos: Se estudiaron 17 cepas de ECUP, aisladas de pacientes adultos hospitalizados en dos instituciones de salud. La susceptibilidad frente a ocho antimicrobianos se determinó por el método de microdilución en caldo (MDC). Las β-lactamasas de espectro extendido (BLEE) y carbapenemasas fueron detectadas fenotípicamente. Los grupos filogenéticos y la detección de los genes de virulencia se determinaron por reacción de polimerasa en cadena. Resultados: Todas las cepas sintetizaban BLEE y de éstas, 41% se asoció a la producción de una carbapenemasa (KPC o MBL). El filogrupo B2 (41%) fue predominante. Los genes de virulencia más frecuentes fueron fimH y fyuA con 82% cada uno. Sólo un aislado clasificado en el filogrupo F fue positivo al conjunto de seis genes estudiados. Discusión: La diversidad de asociaciones entre genes de virulencia y perfiles de resistencia, en las cepas ECUP evolucionan continuamente. Además, su distribución en los distintos grupos filogenéticos depende en gran medida de las características clínico epidemiológicas de los grupos de estudios.

Abstract Background: The difference between the pathogenic isolates and commensals of Escherichia coli is based on their phylogenetic antecedents. In Venezuela there are few studies that describe the pathogenic potential of phylogenetic groups in E. coli. Aims: Relate antimicrobial susceptibility, distribution of phylogenetic groups and virulence genes in strains of uropathogenic E. coli (ECUP) isolated from patients with UTI. Methods: We studied 17 ECUP strains, isolated from adult patients hospitalized in two health institutions. The susceptibility to 8 antibiotics was determined by the broth microdilution (MDC) method. Extended spectrum β-lactamases (ESBL) and carbapenemases were phenotypically detected. The phylogenetic groups and the detection of the virulence genes were determined by PCR. Results: All strains synthesized ESBL and of these, 41% were associated with the production of a carbapenemases (KPC or MBL). The phylogroup B2 (41%) was the most predominant. The most frequent virulence genes were fimH and fyuA with 82% each. Only one strain from group F was positive to the 6 genes studied. Discussion: The diversity of associations between virulence genes and resistance profiles in the ECUP are evolving continuously, their distribution in the different phylogenetic groups depends to a large extent on the clinical epidemiological characteristics of the study groups.

In vitro screening of multidrug resistance uropathogenic Escherichia coli from the urban area of Namakkal district

The aim of the study is to screen the multiple drug resistance (MDR) Uropathogenic Escherichia coli (UPEC) fromthe urban area of Namakkal district. To detect UPEC resistant by using different antibiotics and to analyze the virulentcharacteristics of UPEC and amplification of extended-spectrum beta-lactamases genes by multiplex polymerasechain reaction. Total 450 samples individually collected from the urinary tract infection (UTI) patients’ and directstreaked on to the eosin methylene blue agar plates. Significant growth indicates E. coli. HiCrome UTI agar was usedfor rapid identification of uropathogenic E. coli. Out of 450 samples, only 62 isolates of E. coli were subjected tovirulence characteristics, such as slime production (34%), hemolytic activity (56%), and beta-lactamase production(43%). Antibiotic sensitivity test was performed with 13 different antibiotics. Among them, 62 isolates were E. coli,only five were resistant to 10 antibiotics, possess virulence characteristics. Four strains (E-12, E23, E-58, and E-97)have Temoneira, sulfhydryl variable, and cefotaxime hydrolyzing capabilities (CTX-M) antibiotic resistance genes,and E-07 have only CTX-M gene. As E. coli is the main infectious agent in patients with UTI and a potent pathogen,it was difficult to treat with routine antibiotics because day-by-day microbes are resisting to common drugs. Hence,they need alternative therapy

Detection of various virulence factors in high level aminoglycoside resistance and vancomycin resistant enterococci isolates of uropathogenic Enterococci

Background: Enterococci are common commensal organism of enteric tract and act as opportunistic pathogen and may cause infection in community as well as in hospitalised individuals. In present study association of several types of virulence factors like haemolysin, gelatinase and biofilm formation have been studied among HLAR and Vancomycin resistant Enterococci (VRE) isolates of enterococci among UTI patients.Methods: The samples were collected from all hospitalized and OPD patients of MBS Hospital, JK Lone Hospital and NMC Hospital. Government Medical College, Kota, Rajasthan, India. A total of 360 isolates of enterococcus were collected during the period of 2 years from April 2016 to April 2018 in microbiology laboratory, Department of Microbiology, Government Medical College, Kota, Rajasthan, India. All virulence factors were detected by phenotypic methods and MIC values were detected for high level gentamicin and vancomycin.Results: Among all enterococcal isolates most common factor was biofilm production 191 (53.05%) followed by haemolysin 131 (36.38%) and gelatinase production 72 (20%). Total resistant (MIC> 500 µg/ml) isolates for gentamicin was 194 (89.4%). In agar dilution 14 (11.2%) isolates were found sensitive, 61 (48.8%) isolates were found intermediate and 50 (40%) isolates were found to be resistant for vancomycin. HLAR and VRE was maximum associated with haemolysin + bio-film followed by gelatinase+biofilm, haemolysin+gelatinase+bio- film and least with haemolysin + gelatinase.Conclusions: In present study enterococcus show significant production of biofilm and other virulence factors. With production of biofilm they become more resistant to routinely used concentration of antibiotics posing threat for treatment failure. A continuous monitoring is needed particularly for resistance to aminoglycoside and vancomycin to stop their institutional spread. Judicial use of antibiotics should be encouraged both in community as well as in institutions.

Infecciones del tracto urinario, inmunidad y vacunación / Urinary tract infections, immunity, and vaccination

Resumen Las infecciones del tracto urinario (ITU) se consideran como una de las principales causas de morbilidad en el mundo, y Escherichia coli uropatogénica (UPEC, por sus siglas en inglés) es el agente causal asociado a estas infecciones. La alta morbilidad generada por las ITU y la limitación de tratamientos debido al aumento de la resistencia bacteriana a los diversos antibióticos inducen la búsqueda de nuevas alternativas contra estas infecciones. El conocimiento que se ha generado acerca de la respuesta inmunitaria en el tracto urinario (TU) es importante para el desarrollo de estrategias efectivas en la prevención, el tratamiento y el control de las ITU. Los avances en las herramientas de biología molecular y bioinformática han permitido generar proteínas de fusión consideradas como biomoléculas potenciales para el desarrollo de una vacuna viable contra las ITU. Las adhesinas fimbriales (FimH, CsgA y PapG) de UPEC son factores de virulencia que contribuyen a la adherencia, la invasión y la formación de comunidades bacterianas intracelulares. Pocos estudios in vivo e in vitro han mostrado que las proteínas de fusión promueven una respuesta inmunitaria eficiente y de protección contra las ITU causadas por UPEC. Adicionalmente, la vía de inmunización intranasal con moléculas inmunogénicas ha generado una respuesta en la mucosa del TU en comparación contra otras vías de inmunización. El objetivo de esta revisión fue proponer un diseño de vacuna contra las ITU causadas por UPEC, describiendo el panorama general de la infección, el mecanismo de patogenicidad de la bacteria y la respuesta inmunitaria del huésped.

Abstract Urinary tract infections (UTI) are considered one of the main causes of morbidity worldwide, and uropathogenic Escherichia coli (UPEC) is the etiological agent associated with these infections. The high morbidity produced by the UTI and the limitation of antibiotic treatments promotes the search for new alternatives against these infections. The knowledge that has been generated regarding the immune response in the urinary tract is important for the development of effective strategies in the UTI prevention, treatment, and control. Molecular biology and bioinformatic tools have allowed the construction of fusion proteins as biomolecules for the development of a viable vaccine against UTI. The fimbrial adhesins (FimH, CsgA, and PapG) of UPEC are virulence factors that contribute to the adhesion, invasion, and formation of intracellular bacterial communities. The generation of recombinant proteins from fimbrial adhesins as a single molecule is obtained by fusion technology. A few in vivo and in vitro studies have shown that fusion proteins provide an efficient immune response and protection against UTI produced by UPEC. Intranasal immunization of immunogenic molecules has generated a response in the urinary tract mucosa compared with other routes of immunization. The objective of this review was to propose a vaccine designed against UTI caused by UPEC, describing the general scenario of the infection, the mechanism of pathogenicity of bacteria, and the immune response of the host.

Recurrent Guillain-Barré Syndrome Following Urinary Tract Infection by Escherichia coli

Recurrent Guillain-Barré syndrome (GBS) is a rare, immune-mediated disease of the peripheral nervous system. It has been reported to occur at intervals ranging from four months to 10 years; published case studies suggest that 1%–6% of patients who have had GBS will experience recurrent attacks. The most commonly identified infections coinciding with GBS are Campylobacter jejuni, Haemophilus influenzae, Mycoplasma pneumonia, and cytomegalovirus, while an antecedent infection with Escherichia coli is very uncommon. In this case report, we present a rare episode of recurrent GBS, which followed a urinary tract infection (UTI) by E. coli, and an accompanying literature review. A 75-year-old woman with a prior history of acute motor axonal neuropathy (AMAN), a subtype of GBS, presented with subsequent weakness of limbs and areflexia following 10 days of fever, frequency, and dysuria. Base on nerve conduction studies, cerebrospinal fluid analysis and other clinical investigation, we diagnosed the patient with recurrent GBS caused by E. coli. The patient recovered with mild subjective weakness following treatment of intravenous immunoglobulin with ceftriaxone. We suggest that E. coli causes UTI could be one of the diverse trigger factors involved in recurrent GBS.

Niaogan Prescription protects bladder epithelial cells against uropathogenic Escherichia coli invasion / 中草药

Objective To observe the role of Niaogan Prescription against uropathogenic Escheriehia coli (UPEC) in invading bladder epithelial cells, and to discuss its mechanism of action. Methods After HTB-9 infected with UPEC, the cell invasion model was used to observe the protective effects of drug urine in human bladder cancer cell 5637 (HTB-9) against UPEC invasion. Effects of Niaogan Prescription on the main steps of signaling pathway: Toll-like receptor 4 (TLR4), adenylate cyclase 3 (AC3), cyclic adenosine monophosphate (cAMP), protein kinase A (PKA), and Rac-1 was investigated by molecular biology techniques. Results Compared with the blank urine high dose group (10% volume fraction of blank urine), treatment with the drug-containing urine high dose group of Niaogan Prescription (10% volume fraction of drug urine) resulted in a significant decrease in UPEC invasion, the bacterial invasion rate of the rat blank urine group had no significant difference compared with the model group. Compared with the blank urine high dose group, the drug-containing urine high dose group of Niaogan Prescription improved TLR4 and AC3 protein expression, increased intracellular cAMP content, promoted PKA activation, and inhibited Rac1 activity. Conclusion Niaogan Prescription have the certain capacity against UPEC invasion of bladder epithelial cells, which mechanism is related to TLR4/cAMP signaling pathway. Niaogan Prescription; uropathogenic Escheriehia coli; invasion; human bladder epithelial cells; TLR4/cAMP signal pathway

Relationships between Virulence Factors and Antimicrobial Resistance among Escherichia coli Isolated from Urinary Tract Infections and Commensal Isolates in Tehran, Iran

OBJECTIVES: Uropathogenic Escherichia coli (UPEC) are the major cause of urinary tract infections (UTIs). Here, we determined whether sensitivity to antibiotics was related to the prevalence of iron scavenging genes, or to biofilm and hemolysis formation. METHODS: A total of 110 UPEC and 30 E coli isolates were collected from the urine of UTI patients and feces of healthy individuals without UTI, respectively. The presence of iron receptor genes and phenotypic properties were evaluated by polymerase chain reaction and phenotypic methods, respectively. Susceptibility to routine antibiotics was evaluated using the disc diffusion method. RESULTS: The prevalence of iron scavenging genes ranged from 21.8% (ireA) to 84.5% (chuA) in the UPEC. Resistance to ceftazidime and cefotaxime was significantly correlated with the presence of fyuA and iutA iron genes. Biofilm production was significantly associated with the prevalence of fyuA and hma iron genes. A higher degree of antibiotic resistance was exhibited by isolates that produced biofilms than by their non-biofilm producing counterparts. CONCLUSION: Our study clearly indicates that biofilm production is associated with antibiotic resistance, and that iron receptors and hemolysin production also contribute to reduced antibiotic sensitivity. These results further our understanding of the role that these virulence factors play during UPEC pathogenesis, which in turn may be valuable for the development of novel treatment strategies against UTIs.

Prevalence of ambler class a extended-spectrum- ß -lactamases (ESBLS) among Uropathogenic Escherichia coli strains isolated from Rasht city, Iran / Prevalência de ambler extended-spectrum classe A- ß -lactamases (ESBLs) entre Uropathogenic Escherichia coli isolados da cidade de Rasht, Irã

Extended spectrum -lactamases (ESBLs), a group of bacterial enzymes which are the major cause of resistance to penicillins, broad spectrum cephalosporins and monobactams, are found among the member of Enterobacteriaceae. The class A ESBLs are mainly encoded by the plasmid mediated blaSHV, blaTEM and blaCTX-M genes. In this study, prevalence of Ambler class A ESBL genes among uropathogenic E. coli (UPEC) isolates associated with community acquired infections and their antibiotic susceptibility pattern was investigated. Seventy UPEC strains were isolated from urine samples and subjected to antimicrobial susceptibility assay using disk diffusion method. Phenotypic screening of ESBL production was evaluated according to the CLSI combined disk method. Genotyping of Ambler class A ESBLs was investigated using PCR. According to the results, ESBLs was identified in 37 isolates while molecular assay showed 47 isolates harbored ESBL genes. The most prevalence was recorded for blaTEM (74.2%) followed by blaCTX-M (43.2%) and blaSHV (12.2%). Imipenem was the most effective drug and ESBL producing isolates showed higher resistance to CAZ, CRO, CFZ, CTX and FOX compared to non ESBL isolates. In conclusion, high prevalence of class A ESBL genes was observed in our study which needs more consideration and rational antibiotic prescription.

As -lactamases de espectro alargado (ESBLs), um grupo de enzimas bacterianas que são a principal causa de resistência às penicilinas, cefalosporinas de largo espectro e monobactamas, encontram-se entre os membros das Enterobacteriaceae. As ESBLs de classe A são principalmente codificadas pelos genes blaSHV, blaTEM e blaCTX-M mediados por plasmídeo. Neste estudo, foi investigada a prevalência dos genes ESBL de classe A de Ambler entre isolados de E. coli uropatogênicos (UPEC) e seu padrão de suscetibilidade aos antibióticos. Setenta cepas UPEC foram isoladas a partir de amostras de urina e submetidas a ensaio de susceptibilidade antimicrobiana utilizando o método de difusão em disco. O rastreio fenotípico da produção de ESBL foi avaliado de acordo com o método de disco combinado CLSI. A genotipagem de ESBL de classe A de Ambler foi investigada usando PCR. De acordo com os resultados, as ESBLs foram identificadas em 37 isolados enquanto que o ensaio molecular mostrou 47 isolados portadores de genes ESBL. A maior prevalência foi registrada para blaTEM (74,2%), seguida de blaCTX-M (43,2%) e blaSHV (12,2%). O imipenem foi o fármaco mais eficaz e os isolados produtores de ESBL apresentaram maior resistência a CAZ, CRO, CFZ, CTX e FOX em comparação com os isolados não ESBL. Em conclusão, a alta prevalência de genes ESBL de classe A foi observada em nosso estudo, que necessita de maior atenção e prescrição de antibióticos racionais.

Resistencia bacteriana de Escherichia coli uropatogénica en población nativa amerindia Kichwa de Ecuador / Bacterial resistance of uropathogenic Escherichia coli in native Kichwa Amerindian population of Ecuador

Contexto: Escherichia coli uropatógena (ECUP) se presenta como uno de los principales agentes etiológicos en infecciones del tracto urinario (ITUs) no complicadas (70-95%). El objetivo del tratamiento de ITUs no complicadas es obtener curación clínica y microbiológica. Para ello, es de particular importancia el conocimiento de las tasas de resistencia antibiótica local. Objetivo: identificar los perfiles de resistencia a antibióticos de primera línea para ITUs no complicadas en poblaciones nativas amerindias Kichwas ecuatorianas, en donde el tratamiento empírico se basa en trimetoprim/sulfametoxazol, ampicilina, y ciprofloxacina mayoritariamente. Métodos: se analizaron 335 muestras de orina procedentes de las poblaciones de Zumbahua, Colta y Guamote, en un periodo de 4 meses (febrero-mayo 2016). Las muestras fueron incubadas por 24 y 48 horas en agar Eosin Methylene Blue (EMB), para luego ser identificadas en género y especie por pruebas bioquímicas. Para determinar la susceptibilidad antibiótica, se realizó la técnica de difusión en disco de Kirby-Bauer. Para la Concentración Inhibitoria Mínima (CIM), se utilizó la técnica de microdilución en caldo (Vitek 2). El método de doble disco fue la técnica utilizada para la detección de betalactamasas de espectro extendido (BLEE). Resultados: noventa (26,9%) muestras mostraron un recuento significativo de ≥105 (ufc)/ml, compatibles con ITUs. El microorganismo identificado con mayor frecuencia fue E. coli (n=75; 83,3%). La resistencia antibiótica encontrada para los aislados de E. coli fue de 56,7% a trimetoprim/sulfametoxazol, 52,5% a ampicilina, 43.3% a ácido nalidíxico, 32.5% a ciprofloxacina, 28.3% a norfloxacina, 25% a levofloxacina, 15.85% a cefazolina, 17.5% a cefoxitina, 15% a cefuroxima, 15% a ceftazidima, cefotaxima, y ceftriaxona, 15% a cefepima, 7,5% a nitrofurantoina y 1,7% a fosfomicina. Se identificaron 7 aislados productores de betalactamasas de espectro extendido (BLEE). Conclusión: con los resultados obtenidos se recomienda no utilizar ampicilina, trimetoprim/sulfametoxazol, ni quinolonas en la zona estudiada como terapia empírica. Se sugiere instaurar tratamiento empírico con fosfomicina o nitrofurantoina para ITUs no complicadas. (AU)

Multi-drug resistant uropathogenic Escherichia coli and its treatment by Chinese medicine / 中国结合医学杂志

<p><b>OBJECTIVES</b>To investigate the resistance and virulence profiles of uropathogenic Escherichia coli (UPEC) and its treatment by Chinese medicine (CM) Fuzheng Qingre Lishi Formula (, FQLF).</p><p><b>METHODS</b>UPEC strains were isolated from recurrent urinary tract infections (UTIs) patients. Patient sensitivities to 17 antibiotics were tested by the disk diffusion method. Virulence genes were screened by plolymerase chain reaction. A mouse model was constructed using a multi-drug resistant and virulent UPEC strain and treated with FQLF or the antibiotic imipenem. The treatment efficacy was evaluated by bacterial clearance from urine and the urinary organs.</p><p><b>RESULTS</b>A total of 90 UPEC strains were collected, and 94.4% of the isolates were resistant to at least 1 antibiotic. Approximately 66.7% of the UPEC strains were multi-drug resistant. More than one virulence gene was found in 85.6% of the isolates. The extended-spectrum β-lactamases (ESBL)-positive strains were more resistant than the negative ones. The virulence gene number was positively correlated with the resistance number (P<0.05). A mouse model was successfully constructed using UPEC10. Treatment with either FQLF or antibiotics significantly cleared bacteria from the mouse urine after 14 days. In the untreated control, the bacteria lasted for 28 days. FQLF treatment of the UTI mouse model greatly reduced the bacterial number in the kidney and bladder, but could not completely clear the bacteria.</p><p><b>CONCLUSIONS</b>Multi-drug resistance is common among UPEC isolates, and the resistance is positively related with virulence. FQLF could treat UPEC UTIs, but could not completely clear the bacteria from the host.</p>

Clonal and Virulence Distribution of Uropathogenic Escherichia coli Isolated from Children in Korea

Urinary tract infections (UTI) are one of the most frequent infectious diseases. Uropathogenic Escherichia coli (UPEC) are among major pathogens causing UTI. A variety of virulence genes are mainly responsible for the severity of these emerging infection. This study investigate the influences of virulence properties of UPEC isolates with reference to multi-locus sequence typing (MLST). The aim of this study was targeted that investigation of the bacterial pathogenicity associated with UTI in children. A total of 58 UPEC isolates were collected from urine samples from patients with clinical diagnosis of uncomplicated UTI. The MLST of UPEC strains were assessed by methods based on polymerase chain reaction. Motility was evaluated using soft-agar plates. Biofilm formation was analyzed in microtiter dish biofilm formation assay. Cell death assay was analyzed by Annexin V/Phosphatidylserine staining and DNA fragmentation assay. According the result, the predominant sequence type (ST) was ST95 (24.1%) and ST73 (17.2%). There were some difference in virulence gene and antibiotics resistance between ST95 and ST73. The number of 11 (18.9%) isolates were strongly adherent. Based on the detected biofilm formation, these strongly adherent are almost ST73. The ST95 was higher than ST73 in population, but ST95 was lower than ST73 in motility and cell death induction. This study indicated that the UPEC molecular strains are related to some virulence traits. Furthermore, the virulence factors carried by ST73 strains contribute to their abilities to colonize the host and cause disease.

Progress in the studies of virulent characteristics of uropathogenic Escherichia coli / 中华检验医学杂志

Uropathogenic Escherichia coli ( UPEC) is the most common pathogen causing urinary tract infection ( UTI).Various virulent factors of UPEC involved in the pathogenic process of UTI , including adhesins, iron uptake-related factors, protectins, and toxins.Adhesion factors can help UPEC to adhere to the surface of the epithelial cell in the host urinary tract.Iron ion acquisition factors can help the pathogen to get iron from the host body , and then multiply and cause disease.Various toxins produced by the pathogen can cause damage to the host epithelial cells and release of nutrients and other nutritional factors that help UPEC to survive and grow.Protectins can protect UPEC from the host complement system mediated bactericidal effect and the phagocytosis of phagocytic cells.In this review , the most recent research progress on the various virulence characteristics of UPEC will be discussed.