Comparison of genomic profiles of Escherichia coli isolates from urinary tract infections

Formally included in the larger category of extraintestinal pathogenic Escherichia coli (ExPEC), the uropathogenic E. coli remains the most frequent cause of urinary tract infection (UTI), an important endemic health problem. The genomic DNA of E. coli urinary isolates from adults diagnosed with urinary tract infections and of E. coli fecal isolates from healthy subjects was analysed by PCR for the presence of virulence factor encoding genes pap, sfa/foc, afa, hly and cnf and by field inversion gel electrophoresis (FIGE) fingerprinting of XbaI DNA macrorestriction fragments. The aim was to obtain more detailed microbiological data regarding the community circulating strains in respect of their virulence potential and genetic relatedness. Almost 70% of the urinary strains carried at least one of the target virulence genes, and only 35.5% of the fecal E. coli strains were positive in the PCR screening. Taking into account the virulence genotypes exhibited, a part of the strains isolated from the urinary tract could be defined as belonging to the ExPEC pathotype. A unique FIGE profile was obtained for each of the selected isolates and the dendrogram generated by Taxotron software package analysis suggested a polyclonal population of potential uropathogenic strains clustered into 14 groups of only 60% similarity. For better understanding the epidemiology of UTIs, diseases commonly caused by such a heterogeneous species like E. coli, molecular analysis methods could be essential due to their increased power of identification and fingerprinting.     

Virulence characteristics of Escherichia coli isolates from children with urinary tract infections

The urinary tract is among the most common sites of bacterial infection and E. coli is by far the most common infecting agent in children and adults of both sexes. In an attempt to evaluate the intrinsic virulence of E. coli uroisolates from children, 54 strains were assessed by using PCR for the presence of five representative genetic determinants coding for adherence systems (pap, sfa/foc, afa), and toxins (hly and cnf). The prevalence of pap, sfa/foc and afa genes was 55%, 54%, and 44%, respectively. Hemolysin-encoding gene hly was detected in 55% strains, while cnf was exhibited by 35% of the screened E. coli isolates. Among the 39 PCR positive strains isolated from children's urine cultures the co-occurrence of the various targeted virulence genes was detected in 30 strains, the virulence profiles identified suggesting the presence of their localization on chromosomal regions known as pathogencity-associated islands. The rapid and reliable detection of the intrinsic virulence potential by this molecular approach could be very useful when evaluating the importance of microorganism pathogenicity versus host's susceptibility for developing an overt symptomatology of infection.     

Colicins ofEscherichia coli strains causing urinary tract infections,sensitivity of these strains towards colicins,and their O-serotypes

Out of 175Escherichia coli strains isolated from the urinary tract 45% were colicinogenic. Out of these 175 strains 19% produced colicin V, colicin G was produced by 6% of the strains, colicin I by 9%, colicin A by 4%, colicin B by 4.5%, colicins E by 7.4%, and 1% yielded colicin K. The number of transmissible col factors was 10%. The majority of strains produced colicin V but the sensitivity towards it was also among the highest. The relationship between the type of colicin and the O-serotype, found in some case, may have been caused by strain selection which apparently takes place in hospitalized patients. Serologic typing supplemented by typing of colicins helps in elucidating the epidemiological relationships.     

Reduction in co-trimoxazole resistance inEscherichia coli isolates from urinary tract infections between 1995 and 2005: a multicenter study in Ankara, Turkey

Co-trimoxazole resistance inEscherichia coli isolates from urinary tract infections (UTI) was assessed in 382 strains from 1995 and 510 strains from 2005. The strains were collected from five microbiology laboratories in Ankara, Turkey. Documentation on patient gender, age and outpatient/inpatient status was collected in 2005, but not in 1995. The resistance percentages were 751% in 1995 and 55.5% in 2005. This reduction in resistance percentage was statistically significant, overall in all except two of the participating laboratories. The resistance percentage in 2005 was 61.1% for children (n=208) and 51.2% for adults (n=258), 53.7% for females (n=380) and 60.8% for males (n=130), and 55.3% for outpatients (n=400) and 56.4% for inpatients (n=110). The reduction in resistance is believed to be a consequence of reduced usage. Although decreased, the level of co-trimoxazole resistance remains high, and continued avoidance of its use for empiric treatment of UTI in Turkey appears to be an appropriate strategy.     

Intracellular bacterial communities of uropathogenic Escherichia coli in urinary tract pathogenesis

Urinary tract infections in young, healthy women frequently recur, despite their traditional classification as acute infections. Conventional wisdom dictates that uropathogens causing recurrent infections in such individuals come from the fecal or vaginal flora, in the same manner as the initial infection. However, recent studies of uropathogenic Escherichia coli have found that it can carry out a complex developmental program within the superficial epithelial cells of the mouse bladder, forming intracellular bacterial communities with many biofilm-like properties. These intracellular biofilms allow the bacteria to outlast a strong host immune response to establish a dormant reservoir of pathogens inside the bladder cells. Re-emergence of bacteria from this reservoir might be the source of recurrent infection.     

R factors ofEscherichia coli strains causing urinary tract infections,their types of transfer factors and differences in their transmissibility toCitrobacter andSalmonella typhimurium recipient strains

A total of 107 R factors was demonstrated in 103Escherichia coli strains isolated from the urine of patients with urinary tract infections. Four of the strains were in a hetero-R state, i.e. each cell possessed two R factors with different types of transfer factors. Of the total number of R factors, 76.6% were fi⁺ and 23.4% fi⁻. The recipient strainCitrobacter O-7, 3b, 1c accepted all the given R factors, while the other,Salmonella typhimurium, accepted only 20%. Both the recipient strains accepted fi⁺ and fi⁻ factors. We presume that the transmissibility of R factors depends on their type of transfer factor, but in relation to criteria other than fi⁺ and fi⁻ and the recipient's affinity.     

尿内毒素定量检测对快速诊断革兰阴性菌导致的泌尿道感染的价值研究

目的研究尿内毒素定量检测与培养结果的一致性,及对于快速诊断革兰阴性菌导致的泌尿道感染的价值。方法采用动态浊度法鲎试验定量检测240份尿液中的内毒素含量,同时进行常规尿培养和鉴定,SPSS 16.0分析检测结果,采用t检验比较不同培养结果之间内毒素检测结果差异的显著性,Kappa值评价内毒素检测和培养结果的一致性。结果培养结果为革兰阴性菌生长的尿液标本,与革兰阳性菌或真菌生长及培养2 d无菌生长的标本内毒素定量检测结果的差异有统计学意义(t=11.98,t=12.00,均为P0.001);革兰阳性菌或真菌生长的尿液标本与培养2 d无菌生长的标本内毒素定量检测结果的差异无统计学意义(t=1.38,P=0.183)。结论尿内毒素定量检测对于快速诊断革兰阴性菌导致的泌尿道感染有着较好的应用前景。     

Interaction between atypical microorganisms and E. coli in catheter-associated urinary tract biofilms

Most biofilms involved in catheter-associated urinary tract infections (CAUTIs) are polymicrobial, with disease causing (eg Escherichia coli) and atypical microorganisms (eg Delftia tsuruhatensis) frequently inhabiting the same catheter. Nevertheless, there is a lack of knowledge about the role of atypical microorganisms. Here, single and dual-species biofilms consisting of E. coli and atypical bacteria (D. tsuruhatensis and Achromobacter xylosoxidans), were evaluated. All species were good biofilm producers (Log 5.84–7.25 CFU cm^?2 at 192?h) in artificial urine. The ability of atypical species to form a biofilm appeared to be hampered by the presence of E. coli. Additionally, when E. coli was added to a pre-formed biofilm of the atypical species, it seemed to take advantage of the first colonizers to accelerate adhesion, even when added at lower concentrations. The results suggest a greater ability of E. coli to form biofilms in conditions mimicking the CAUTIs, whatever the pre-existing microbiota and the inoculum concentration.     

The role of Ureaplasma urealyticum in urinary tract infections. Demonstration of antibodies against ureaplasmas in the urine

This study reports the presence of antibodies against Ureaplasma urealyticum in the urine in 17.2% (85) of 493 patients with chronic pyelonephritis. At the same time in 37 of antibody-positive urine samples evidence was found of cultivated Ureaplasma urealyticum, other bacterial germs were isolated infrequently. These culturally and serologically positive urine samples mostly showed pathologic findings of urine sediments. These results led us to believe in a possible involvement of the Ureaplasmas in keeping inflammation processes in the upper urinary tract.     

尿液菌落计数在儿童泌尿感染筛查中的应用

目的检验和比较尿液菌落计数在筛选儿童泌尿系统感染中的可靠性。方法收集儿童中段尿液,10μL进行细菌培养和菌落计数,剩余样本进行干化学分析,主要进行沉渣镜检、亚硝酸盐和白细胞酯酶检测。结果共检测中段尿液标本112份,其中32例培养出有意义细菌,阳性率为28.6%。尿沉渣镜检、干化学分析的敏感性分别为68.8%和78.1%,特异性分别为92.5%、90.0%。另有7例培养阳性,干化学分析阴性,假阴性为21.9%;6例沉渣镜检阳性,培养阴性,假阳性率7.5%;沉渣镜检、干化学分析的阳性预示值分别为78.6%和75.8%。结论尿液细菌培养和菌落计数在筛选儿童泌尿感染的敏感性要明显优于尿沉渣镜检和干化学分析,在儿童泌尿系统感染筛查中起到重要作用。尿沉渣镜检和干化学分析因为敏感性不足、结果易受到感染类型影响。尿液菌落计数可作为儿童泌尿系统感染检测的重要指标。