Sfa Genes Research Articles

Expression of sfa and afa Genes in Uropathogenic Escherichia coli Under Probiotic Effect

Expression of sfa and afa Genes in Uropathogenic Escherichia coli Under Probiotic Effect

Public health concern of antimicrobial resistance and virulence determinants in E. coli isolates from oysters in Egypt

The emergence of critical-priority E. coli, carrying a wide array of resistance and virulence factors through food sources, poses a significant challenge to public health. This study aimed to investigate the potential role of oysters sold in Egypt as a source for E. coli, identify their resistance and virulence-associated gene profiles, and assess associated zoonotic risks. A total of 33 pooled fresh oyster samples were obtained from various retail fish markets in Egypt and examined bacteriologically for the presence of E. coli. Antimicrobial resistance was performed by the disk-diffusion method, and the multiple antibiotic resistance index (MAR) was calculated. All isolates were screened for extended-spectrum beta-lactamase (ESBL) (blaTEM, blaSHV, blaCTX−M, and blaOXA−1), plasmid-mediated AmpC blaCMY−2, and carbapenemases (blaKPC, blaNDM, blaVIM, and blaOXA−48) genes by Polymerase chain reaction. Moreover, the presence of virulence-encoding genes was investigated. The virulent MDR strains were clustered using R with the pheatmap package. The prevalence of E. coli was 72.7% (24 out of 33), with 66.7% of the isolates classified as multi-drug resistant, and 75% exhibited MAR values exceeding the 0.2 threshold. Different antimicrobial sensitivity phenotypes and genotype profiles were identified in E. coli isolates. The most prevalent gene detected among all isolates was blaTEM (22/24, 91.7%). Notably, all non-ESBL producers were positive for blaCMY2. Carbapenem-resistant and carbapenem-intermediate strains were carbapenemase producers, with the predominance of the blaKPC gene (11/24, 45.8%). Remarkably, twelve out of sixteen virulence genes were identified, with papC (21/24, 87.5%) and sfa (16/24, 66.7%) genes being the most prevalent. Most isolates carry virulence genes primarily associated with extra-intestinal pathogenic E. coli (ExPEC) (87.5%) and enteropathogenic (EPEC) (70.8%) pathotypes. Four E. coli isolates exhibit cluster patterns. This study provides the first insight into the emergence of virulent MDR E. coli among oysters in Egypt. It underscores the potential role of oysters as a source for disseminating these strains within aquatic ecosystems, presenting a possible threat to public health.

The presence of virulence factor genes downregulates uterine AQP3 and alters glutathione peroxidase activity and uterine histopathology in canine pyometra.

Present study was designed to evaluate the role of virulence factor genes (papG, cnf1 and hylA) in the pathogenesis of canine pyometra. Antimicrobial susceptibility test and detection of virulence genes were performed Escherichia coli (E. coli) detected in uterine swab samples. Animals were divided into two groups based on the presence (VF+, n:14) or absence (VF-, n:7) of the virulence factor genes papG, cnf1 and hylA. Blood and tissue glutathione peroxidase activity, uterine histopathologic analysis and AQP3, ESR1, PGR, OXTR gene expressions were determined in both groups. Statistical analyses were performed using Stata version 15.1. All E. coli isolates were susceptible to amikacin, whereas resistant to ampicillin, amoxicillin/clavulanic acid and lincomycin. None of the isolates were susceptible to cefotaxime. E. coli isolates had at least one virulence gene. The most prevalent gene was fimH (100%), followed by fyuA (95.8%), usp (83.3%), sfa (75%), cnf1 and hlyA (70.8%) genes. Blood GPx activity was greater in VF+ animals. On the other hand, uterine tissue GPx activity was lower in VF+ group compared to the control group. Expression levels of AQP3 were upregulated more than fivefold in VF-dogs compared to the control group. In addition, AQP3 expression levels were found approximately threefold higher in VF (-) than VF (+) group (p < .05). Varying degree of inflammation noted for all animals with pyometra, but the presence of bacteria noted only in VF+ animals. In conclusion, the presence of virulence factor genes does not play a role in the histopathological degree of inflammation, the presence of bacteria was found to vary. Serum GPx activity increased in VF+ animals. While the hormone receptor expressions were similar, AQP expression was upregulated in the absence of virulence factor genes.

Effect of thymol on antimicrobial susceptibility, and adhesion genes expression of uropathogenic Escherichia coli isolated from pediatric urinary tract infection

Uropathogenic Escherichia coli (UPEC) is a common cause of urinary tract infections (UTI) in children and currently is one of the leading medical problems. Due to the increase in antibiotic resistance rate, herbal medicines with lower side effects were considered. This study aimed to identify the afa, fimH, and sfa genes of UPEC bacteria isolated from pediatric UTI to investigate the effect of the thyme on the expression of fimH gene. In this cross-sectional study, 160 UPEC were isolated from pediatric UTIs. An antibiotic susceptibility test was performed on six families of antibiotics, including beta-lactams, quinolones, aminoglycosides, carbapenems, sulfonamides, and nitrofurantoin. The micro-broth dilution method was used to determine MIC of thymol. The biofilm production ability of isolated strains was quantified by the microtiter plate method. The PCR technique was used to detectfimH, afa, and sfa adhesion genes, and real-time PCR was used to measure the fimHgene expression. The results of the antibiogram showed that the lowest and highest resistance related to meropenem and imipenem (zero), and 72.5% for cephalothin. MIC showed 80.7% of the isolates were sensitive to thymol. The biofilm production results showed that 3.12%, 53.75%, and 43.12% of the isolates were strong, weak, and no-biofilm (Zero) producers, respectively. After thymol treatment, 26.25% and 73.75% of isolates were weak and no-producer (Zero) biofilms, respectively and there was a significant correlation (P-value=0.042) compared to the control group. The frequency of fimH, sfa, and afa genes was 53.1%, 49.4%, and 29.4%, respectively. The expression of fimHgene after 48h thymol treatment decreased significantly (P-value< 0.05). Due to the significant effects of thymol in preventing the expression of the adhesion gene (fimH) of UPEC bacteria, our study is a proof-of-concept study evaluating bacterial sensitivity to Thymol and its effect on biofilm production invitro. Given the demonstrated promising results of Thymol's effectiveness and the increase in bacterial antibiotic resistance, further studies should be undertaken to determine the safety and effectiveness of Thymol use in the clinical treatment of urinary tract infection. We believe that Thymol may prove to be an effective adjunct to the treatment of bacterial urinary tract infections.

Escherichia coli Strains Responsible for Cystitis in Female Pediatric Patients with Normal and Abnormal Urinary Tracts Have Different Virulence Profiles.

The role of uropathogenic Escherichia coli (UPEC) in colonization and infection of female patients with anatomical and functional abnormalities of the urinary system is elusive. In this study, the phenotype, genotype and the phylogeny of UPEC strains isolated from the urine of pediatric female patients with cystitis of normal and abnormal urinary tract were determined. Multiplex PCR results demonstrated that 86% of the strains isolated from female patients with normal urinary tract (NUT), belonged to the phylo-groups B2 and D. Their prevalence decreased to 23% in strains isolated from patients with abnormal urinary tract (AUT). More of the isolates from AUT patients produced a biofilm on polystyrene and polyvinyl chloride (PVC), adhered to epithelial cells, and encoded pap and sfa genes than strains isolated from female patients with NUT. In contrast, a higher number of hemolysin-producing strains with serogroups associated with UPEC were isolated from patients with NUT. In summary, the results suggest that cystitis in female patients with NUT is associated with ExPEC, whereas cystitis in female patients with AUT is associated with pathogenic intestinal E. coli strains that have acquired the ability to colonize the bladder.

Correlation Between Antimicrobial Resistance, Virulence Determinants and Biofilm Formation Ability Among Extraintestinal Pathogenic Escherichia coli Strains Isolated in Catalonia, Spain.

Escherichia coli is a well-characterized bacterium highly prevalent in the human intestinal tract and the cause of many important infections. The aim of this study was to characterize 376 extraintestinal pathogenic E. coli strains collected from four hospitals in Catalonia (Spain) between 2016 and 2017 in terms of antimicrobial resistance, siderophore production, phylogroup classification, and the presence of selected virulence and antimicrobial resistance genes. In addition, the association between these characteristics and the ability to form biofilms was also analyzed. The strains studied were classified into four groups according to their biofilm formation ability: non-biofilm formers (15.7%), weak (23.1%), moderate (35.6%), and strong biofilm formers (25.6%). The strains were highly resistant to ciprofloxacin (48.7%), trimethoprim-sulfamethoxazole (47.9%), and ampicillin (38%), showing a correlation between higher resistance to ciprofloxacin and lower biofilm production. Seventy-three strains (19.4%) were ESBL-producers. However, no relationship between the presence of ESBL and biofilm formation was found. The virulence factor genes fimH (92%), pgaA (84.6%), and irp1 (77.1%) were the most prevalent in all the studied strains. A statistically significant correlation was found between biofilm formation and the presence of iroN, papA, fimH, sfa, cnf, hlyA, iutA, and colibactin-encoding genes clbA, clbB, clbN, and clbQ. Interestingly, a high prevalence of colibactin-encoding genes (19.9%) was observed. Colibactin is a virulence factor, which interferes with the eukaryotic cell cycle and has been associated with colorectal cancer in humans. Most colibactin-encoding E. coli isolates belonged to phylogroup B2, exhibited low antimicrobial resistance but moderate or high biofilm-forming ability, and were significantly associated with most of the virulence factor genes tested. Additionally, the analysis of their clonal relatedness by PFGE showed 48 different clusters, indicating a high clonal diversity among the colibactin-positive strains. Several studies have correlated the pathogenicity of E. coli and the presence of virulence factor genes; however, colibactin and its relationship to biofilm formation have been scarcely investigated. The increasing prevalence of colibactin in E. coli and other Enterobacteriaceae and the recently described correlation with biofilm formation, makes colibactin a promising therapeutic target to prevent biofilm formation and its associated adverse effects.

GENOTYPIC DETECTION OF THE VIRULENCE FACTORS OF UROPATHOGENIC ESCHERICHIA COLI (UPEC) STRAINS ISOLATED FROM PREGNANT FEMALES AND THEIR CORRELATION WITH ANTIBIOTIC RESISTANCE PATTERN

Urinary Tract Infections (UTIs) during pregnancy are among the most common infections worldwide and can lead to poor perinatal and maternal outcomes, especially in developing countries and requires medical treatment as soon as it is detected. Virulent and resistant strains of Escherichia coli are the most important causative agents for UTIs. The present investigation aimed to detect virulence factors of Uropathogenic Escherichia coli (UPEC), including pap, fim, sfa, aer and hly genes in isolates collected from symptomatic and asymptomatic pregnant females by multiplex polymerase chain reaction (PCR) assay and determine their correlation with antibiotic resistance patterns. Two hundred urine samples collected from pregnant females with or without symptoms of UTI were admitted to antenatal clinic at Obstetrics and Gynecology Department of Al- Zahraa University Hospital in Cairo. Out of 200 cultured urine samples, the cultures that showed ≥ 105 CFU/ml were identified to have significant growth. Found significant bacteriuria was detected in 61 samples and these samples selected for further analysis in this study, the overall prevalence of UTI among pregnant females was (30.5%). Symptomatic pregnant females with UTI were (48.5%) more than asymptomatic (11.3%). The most frequently isolated species was S. saprophyticus (35.0%),followed by E. coli (26.2%),S .aureus (19.4%), C .albicans (5.8%),S. epidermidis (2.9%) and each of K. pneumonia, K .oxytoca, P .vulgaris, E.fecalis, Bacillus spp were(1.9%),while P. auroginosa had the least percentage of isolation (1.0%). Antimicrobial susceptibility pattern was done for 27 UPECisolates showed the highest level of resistance (100%) against β-lactams as (Ampicillin, Penicillin and Ceftriaxone), nitrofurantoin, and ampicillin/sulbactam, while the lowest level of resistance (66.7%) against Cotrimoxazole. Multiple drug resistance (resistance to two or more drugs) was observed in (100%) of the UPEC isolates. The higher level of resistance to antibiotics was observed in symptomatic bacteriuria more than asymptomatic. Among two predominant uropathogens, E. coli showed complete resistance up to 5 different antibiotics of total 11 antibiotics (45.5%) while S. saprophyticus showed complete resistance (18.2%), so E. coli showed higher virulence and resistance than S. saprophyticus to cause UTI, so subjected to Multiplex PCR to detect its virulence factors, which included pap, fim, sfa, aer and hly genes Theadhesive fimH gene was the most frequent in UPEC isolates (92.6%) . These virulence genes detected from total UPEC isolates of symptomatic cases were higher than asymptomatic. On the other hand, UPEC strains carrying the virulence genes were more resistant to the antibiotics used, so these observations confirm the important role of virulence genes of UPEC in in existence of symptoms of UTI and the drug resistance.

Phylogenetic groups and virulence genes of Escherichia coli strains isolated from the children gut microbiota.

Escherichia coli is characterized by a wide intraspecific diversity. The species includes both commensals and pathogens that cause diarrhea and extra-intestinal diseases. Pathogenic strains differ from non-pathogenic ones by the presence of virulence factors and their genes. The phylogenetic structure of the species is represented by four main groups (A, B1, B2, D), which differ in their prevalence among residents of different geographical regions. Pathogenic members of the species have been studied in detail, while non-pathogenic strains have not received such attention. This report presents the results of a study of 511 E. coli strains isolated from the gut microbiota of children without diarrhea and urinary tract infections, aged from 1 month to 17 years, living in St. Petersburg. The main phylogenetic groups were determined by PCR, and E. coli virulence genes associated with diarrhea and extra-intestinal diseases were identified. Results: population structure of E. coli is represented by the following groups: A-33.3%, B1-6.7%, B2-34.0%, D-26%. In the studied population 2.5% of strains belonded to EPEC and 4.5% to EAggEC. EPEC virulence genes were more often detected in strains of phylogroup B1, and EAggEC virulence genes in isolates of phylogroup D. The prevalence of extra - intestinal virulence genes was as follows: pap - 29.5%; sfa - 19.8%; afa - 3.3%; hly - 20.9%; cnf - 17.4%; aer-20.0%. The pap, sfa, hly, and cnf genes were detected mostly in the B2 phylogenetic group. Obtained data shows the similarity of E. coli phylogenetic groups structure in St. Petersburg with E. coli populations isolated from residents of Paris and Sydney. Analysis of the virulence genes prevalence showed the dependence of their presence on the genetic background bacteria.

Klinik E.coli İzolatlarında Virulans Faktor Genlerinin ve Geniş Spekturumlu Beta Laktamazların Araştırılması

The aim of this study was to investigate the presence of pap, hly, cnf, sfa, aer and afa virulans genes and extended-spectrum beta lactamase resistance genes in 83 clinical E.coli isolates. In 83 E. coli isolates, sfa, pap, hly, cnf, aer and afa virulence genes and TEM-SHV, CTX-M1 and CTX-M9 wide spectrum beta-lactamase genes were investigated by polymerase chain reaction (PCR) method. Resistance rates of 83 isolates to cefixim, trimethoprim/sulfamethoxazole, piperacillin/tazobactam, amoxicillin/clavulanic acid, ampicillin, ceftriaxone, cefuroxime, cefuroxime axetil, ciprofloxacin, fosfomycin, gentamicin and ceftazidime antibiotics were 65%, 54%, 19%,38, 74%, 75%, 72%, 75%, 68%, 2%, 30%, 49%, respectively. CTX-M1 was the most common extended spectrum beta-lactamase (ESBL) among the ESBL genes investigated in 51 of 83 strains. TEM type beta-lactamase was detected in 33 strains, CTX-M9 and SHV 1 isolates. The most common virulence gene was aer gene and detected in 40 strains, followed by the cnf gene detected in 15 of the isolates. The pap, hly, afa and sfa genes were determined in 15.6%, 12.04%, 9.6% and 15.6% of the isolates, respectively. In addition, 10 different virulence factor gene patterns were observed in 83 E.coli isolates. Virulence gene combinations were observed in 27 strains of aer, 6 strains of cnf-sfa-pap, 1 strains of aer-pap, 2 strains of aer-afa, 6 strains of afa, 3 strain of sfa, 9 strain of aer-cnf-hly, 4 isalates of sfa-pap, 1 strain of hly-pap and 1 strains of pap genes. In conclusion, investigating bacterial pathogenicity associated with UTI will contribute to better medical intervention.

SF-Assemblin genes in Paramecium: phylogeny and phenotypes of RNAi silencing on the ciliary-striated rootlets and surface organization

BackgroundCilia emanate from basal bodies just underneath the cell membrane. Basal bodies must withstand torque from the ciliary beat and be appropriately spaced for cilia to beat in metachronal waves. Basal body rootlets provide stability for motile cilia. Paramecium has three. Our focus is on the largest one, the striated rootlet (SR). Paramecium basal bodies align in straight rows. Previously we found a potential role for the SR in this alignment. Here we present a phylogeny of the Paramecium homologs of the SF-Assemblin gene of the SR of Chlamydomonas, and the organization of these genes. We describe the phenotypes from RNA interference (RNAi) silencing of genes and gene groups.MethodsPhenotypes of the RNAi depletions were characterized by immunofluorescence (IF), electron microscopy, and mass spectrometry.ResultsWe found 30 genes for Paramecium SF-Assemblin homologs (SFA) organized into 13 Paralog Groups (further categorized in five Structural Groups). Representatives of Paralog Groups were found in the SRs. Silencing the transcripts of any of the Structural Groups correlates with misaligned rows of basal bodies, SRs, and cortical units. The silencing of Structural Groups was key and gave us the ability to systematically disrupt SR structures and cell surface organization.ConclusionsSilencing of SFA genes and Paralog Groups shows no effects on the SR or the cell surface organization. Silencing of the larger Structural Groups has an enormous impact on rows of basal bodies, SRs and cortical units, and SR striations, and length. Misaligned basal bodies have cilia causing the cells to swim in abnormal paths.

Evaluation of pap and sfa Genes Relative Frequency P and S Fimbriae Encoding of Uropathogenic Escherichia coli Isolated from Hospitals and Medical Laboratories; Yasuj City, Southwest Iran

Background: Urinary tract infections (UTI) are one of the common infections worldwide. Escherichia coli is the most common bacteria causing UTI. Objectives: The current study aimed at investigating the susceptibility pattern, biofilm formation, and the relationship between pap and sfa virulence genes and biofilm formation in E. coli strains isolated from patients with UTI from 2017 to 2018 in Yasuj City, Iran. Methods: In the current cross-sectional study, 130 E. coli strains were isolated from patients with UTI. Antimicrobial susceptibility testing was performed by disk diffusion method. The presence of pap and sfa genes was investigated by multiplex polymerase chain reaction (PCR). Biofilm formation was tested using microtiter plate assay and ELISA (enzyme-linked immunosorbent assay) reader for E. coli isolates. Results: Out of 130 E. coli strains isolated from UTI samples, 98 (75.38%) were positive for the biofilm formation, (14% strong, 31% moderate, and 53% weak). The pap and sfa genes were detected in 50% and 29% of the isolates, respectively. Majority of the isolates were resistant against ampicillin (80%) and susceptible to amikacin and meropenem (100%); the pap and sfa genes were detected in 92.85% and 85.71% of the biofilm-forming isolates, respectively. These genes in Uropathogenic E. coli (UPEC) have a significant relationship with strong biofilm formation (P < 0.0001). Conclusions: The obtained results indicated that the frequency of pap gene was higher than that of sfa in E. coli strains isolated from patients with UTI in Yasuj. The pap and sfa genes in E. coli strains had a significant relationship with strong biofilm formation. In addition, the frequency of biofilm formation in antibiotic-resistant E. coli was higher than that of antibiotic-sensitive strains. According to the results of the current study, it is recommended to administer Amikacin or Meropenem in Yasuj to treat UTI.

Characterization of uropathogenic ESBL-producing Escherichia coli isolated from hospitalized patients in western Algeria.

The aim of this study is to assess the prevalence and molecular characterization of uropathogenic Extended spectrum β-lactamases (ESBLs) producing Escherichia coli. During 3 years, all hospitalized patients at the University-affiliated hospital of Tlemcen and presenting urinary tract infections caused by E. coli were considered as potential study participants. These E. coli isolates were examined phenotypically for ESBL production. ESBL strains were subjected to antimicrobial susceptibility testing and were investigated for the presence of plasmid mediated quinolone resistance genes, 16SrRNA methylase genes and virulence genes by using conventional PCR and DNA sequencing. The molecular characterization of ESBL strains was established by phylogenetic grouping method and ERIC-PCR. The overall prevalence of ESBL was 32.5%. The blaCTX-M-15 was the most frequently detected in ESBL isolates, followed by blaCTX-M-14, blaCTX-M-28, blaCTX-M-1 and blaSHV-12 respectively. The plasmid-mediated quinolone resistance genes were detected in the 15 ESBL strains with the aac(6')-Ib-cr gene was the most detected followed by qnrB1 and qnrA1 gene respectively. Among the 22 ESBL isolates resistant to gentamicin and amikacin, the 16SrRNA methylase genes were detected in 4 isolates. The sfa and pap virulent genes were founds in 26% and 22% of isolates receptively. The genotyping analysis of all strains revealed that almost were belonged to phylogenetic groups A1 and A0 and fourteen distinct clones. The emergence of uropathogenic ESBL isolates and the high rate of blaCTX-M are alarming in Algeria. Strict measure must be required to control the further spread of these strains in Algerian hospitals.

Forest restoration approaches affect soil compositions of lignin, substituted fatty acids, and lignin degradation-associated genes

The study of soil microorganisms and the composition of organic carbon is important for understanding the dynamics of soil organic carbon. The means by which forest restoration approaches might affect the soil composition of lignin, substituted fatty acids,11substituted fatty acids, SFA and the lignin degradation genes, remain understudied. We hypothesized that the soils of naturally restored forests have more abundant and stable lignin, SFA, and lignin degradation genes than those of artificially restored plantations. We analyzed the compositions of lignin, SFA, and the lignin degradation genes in soils restored with natural secondary forests, native species plantations, and introduced species plantations. We found that the extractable lignin content, leaf-derived SFA, leaf- to root-derived SFA ratio, and lignin oxidation state were higher in natural secondary forests than in plantations. The abundances of genes involved in glyoxal oxidase, lignin peroxidase, and manganese peroxidase were higher in the soils of natural secondary forests than in those of the two plantation types. No significant differences in the above indices were found between soils of native and introduced plantations. Nutrients and lignin oxidation state, including tree richness, litter stock, available nitrogen, combined clay and silt content, syringyl-based lignin to vanillyl-based lignin ratio, syringic acid to syringaldehyde ratio, and SFA content significantly affected the composition of genes involved in lignin degradation. Nutrients and lignin oxidation state explained 41.9% of the variation in the composition of lignin degradation genes. The lower lignin oxidation state, higher nutrient availability, and higher aboveground inputs led to a higher abundance of lignin degradation genes in the soils of natural secondary forests than in those of the plantations. These results provide insight into the effects of forest restoration approaches on soil carbon sequestration.

Determination of Adhesion Encoding Genes of Uropathogenic Escherichia coli

Background: The bacterial strains which cause symptomatic urinary tract infections possess diverse distinctive properties that enable them to overcome local host defences. The present study was undertaken to screen for the virulence factors of uropathogenic Escherichia coli (UPFC) by molecular methods and to detect the genotypic pathogenicity related sequences pap, sfa, and afa genes in E. coli strains by polymerase chain reaction (PCR). Methods: A total of 350 E.coli isolates were obtained from symptomatic cases of urinary tract infections (UTI) and 60 E. coli faecal isolates were collected from apparently healthy individuals (during February 2014-2016) from outpatient department of a tertiary care hospital in Kerala, India. Isolates were tested for adhesive genes pap, sfa, and afa. Results: Among the UPEC samples, 255 (72.85%) of the isolates were from females and 95 (27.14%) of them were obtained from males. The prevalence of pap gene was seen in 72 of the total isolates which were the highest. The sfa genes were shown by 56 isolates. The afa genes were also in 7 of the isolates. Diversity of the UPEC associated virulence genes were established in this study. Conclusions: The E.coli causing UTI had greater percentage of virulence associated adhesin genes compared to E.coli from control stool samples.

The frequency of pap and sfa genes among Escherichia coli strains isolated from hospitalized patients of Rouhani hospital in babol, Iran

The frequency of pap and sfa genes among Escherichia coli strains isolated from hospitalized patients of Rouhani hospital in babol, Iran

English

This study aimed to identify some important virulence factors, including pap, fim, sfa, aer and hly genes, typical of uropathogenic Escherichia coli (UPEC) in isolates collected from diarrheic and urinary tract infected patients in Khartoum State by multiplex polymerase chain reaction (PCR) assay. A total of 100 clinical specimens (50 urine and 50 diarrhea) were collected. Samples were cultured and identified by conventional method. Most study population were females 57/100 (57%); 42 suffering from urinary tract infections (UTIs) and 15 from diarrhea, while males were 43/100 (43%); 8 suffering from UTIs and 35 from diarrhea. Among enrolled subjects, 83 were positive for one or more uropathogenic E. coli virulent genes, while 17 isolates were negative for all genes. The results of multiplex PCR revealed that thirty two (n=32) diarrheal samples and fourteen (n=14) urine samples were aer positive. Thirty three (n=33) urine samples and eight (n=8) diarrheal samples appeared as fim positive. The genes pap and hly were found in 24 and 14 urine samples, respectively and in 9 and 3 diarrheal samples, respectively, while sfa gene was detected only in 15 urine specimens. The study concluded that fim gene was highly prevalent among UTI patients while aer gene was highly prevalent among diarrhea patients. &nbsp; Key words: Uropathogenic Escherichia coli, fimH, aer, pap, sfa, hly, Sudan.

Genotypic detection of the virulence factors of uropathogenic Escherichia coli isolated from diarrheic and urinary tract infected patients in Khartoum State, Sudan

This study aimed to identify some important virulence factors, including pap, fim, sfa, aer and hly genes, typical of uropathogenic Escherichia coli (UPEC) in isolates collected from diarrheic and urinary tract infected patients in Khartoum State by multiplex polymerase chain reaction (PCR) assay. A total of 100 clinical specimens (50 urine and 50 diarrhea) were collected. Samples were cultured and identified by conventional method. Most study population were females 57/100 (57%); 42 suffering from urinary tract infections (UTIs) and 15 from diarrhea, while males were 43/100 (43%); 8 suffering from UTIs and 35 from diarrhea. Among enrolled subjects, 83 were positive for one or more uropathogenic E. coli virulent genes, while 17 isolates were negative for all genes. The results of multiplex PCR revealed that thirty two (n=32) diarrheal samples and fourteen (n=14) urine samples were aer positive. Thirty three (n=33) urine samples and eight (n=8) diarrheal samples appeared as fim positive. The genes pap and hly were found in 24 and 14 urine samples, respectively and in 9 and 3 diarrheal samples, respectively, while sfa gene was detected only in 15 urine specimens. The study concluded that fim gene was highly prevalent among UTI patients while aer gene was highly prevalent among diarrhea patients. Key words: Uropathogenic Escherichia coli, fimH, aer, pap, sfa, hly, Sudan.

Escherichia Coli: Characteristics of Carbapenem Resistance and Virulence Factors

ABSTRACT In this study, fifty Escherichia coli strains were analyzed by multiplex polymerase chain reaction for the genes expressed carbapenemase and virulence factors in order to determine the presence of carbapenemase and nine virulence factors and investigate the association between these two characteristics. When carbapenemase susceptibility was taken into consideration, OXA-48 type carbapenemase was determined for 22% of the total strains. Also, the frequency of virulence gene regions in E.coli infections and virulence gene profiles of these isolates were examined and the frequency of pap, afa, sfa, fimA, iroN, aer, iutA, hly and cnf-1 genes were 24, 38, 20, 84, 28, 90, 92, 10 and 34% respectively. A significant correlation was found between the presence of fimA and afa gene regions and carbapenem susceptibility (P< 0.05). Based on the combination of carbapenemase and virulence factor genes, 24 different gene profiles were determined for all strains. The results of the study appear to indicate that fimA and afa genes correlate with carbapenem susceptibility, the relations of fimA with urinary tract infections and pap with complicated urinary tract infections. It also indicates that sfa and afa genes correlate with other infections except urinary tract infections.

Antimicrobial resistance and molecular characterization of virulence genes, phylogenetic groups of Escherichia coli isolated from diarrheic and healthy camel-calves in Tunisia.

This study was conducted to determine the prevalence of virulence genes, serogroups, antimicrobial resistance and phylogenetic groups of Escherichia coli strains isolated from diarrheic and healthy camel calves in Tunisia. From 120 fecal samples (62 healthy and 58 diarrheic camel calves aged less than 3 months), 70 E. coli isolates (53 from diarrheic herds and 17 from healthy herds) were examined by PCR for detection of the virulence genes associated with pathogenic E. coli in animals. A significantly greater frequency of the f17 gene was observed in individual camels and in herds with diarrhea, this gene being found in 44.7% and 41.5% of isolates from camels and herds with diarrhea versus 22.5% and 11.7% in camels (p=0.05) and herds without diarrhea (p=0.02). The aida, cnf1/2, f18, stx2 and paa genes were found only in isolates from camels with diarrhea, although at a low prevalence, 1.8%, 3.7%, 1.8%, 3.7% and 11.3%, respectively. Prevalence of afa8, cdtB, eae, east1, iroN, iss, kpsMTII, paa, sfa, tsh and papC genes did not differ significantly between herds with or without diarrhea. Genes coding for faeG, fanC, f41, estI, estII, CS31a and eltA were not detected in any isolates. All isolates were sensitive to amikacin, chloramphenicol, ciprofloxacin, gentamicin and ceftiofur and the highest frequency of resistance was observed to tetracycline, and ampicillin (52.8% and 37.1% respectively). The phylogenetic groups were identified by conventional triplex PCR. Results showed that E. coli strains segregated mainly in phylogenetic group B1, 52.8% in diarrheic herds and 52.9% in healthy herds.

Biofilm formation, antimicrobial susceptibility, serogroups and virulence genes of uropathogenic E. coli isolated from clinical samples in Iran.

BackgroundUropathogenic Escherichia coli O- Serogroups with their virulence factors are the most prevalent causes of UTIs. The present research performed to track common uropathogenic E.coli serogroups, antibiotic resistance pattern of strains and prevalence of virulence genes in isolations having the ability to constitute biofilm.MethodsIn this research 130 E.coli isolation from patients having UTI symptoms were collected and antimicrobial resistance pattern was performed by Kirby-Bauer method. Polymerase chain reaction was done using primer pairs to identify common serogroups of uropathogenic E.coli and studying virulence genes in isolations creating biofilm.ResultsAmong 130 E.coli isolates, 80 (61.53 %) were able to make biofilm that 15 isolates (18.75 %) indicated strong reaction, 20 (25 %) of medium and 45 (56.25 %) of weak biofilm reaction. Among isolations creating biofilm, the highest resistance reported to Ampicillin (87.5 %) and the lowest to Nitrofurantoin (3.75 %). The frequency of fimH, pap, sfa and afa genes in isolations having the ability to create strong biofilm reported 93.33 %, 86.66 %, 86.66 % and 66.66 %, respectively.ConclusionsThe findings indicated the importance of virulence genes in serogroups producing uropathogenic E.coli biofilm. It is recommended that strains producing biofilm before antibiotic use should be studied.