Clinical and molecular epidemiology of quinolone-resistant Escherichia coli isolated from urinary tract infection

Since the use of fluoroquinolone antibiotic in clinical practice was introduced about a decade ago, quinolone-resistant E. coli (QREC) strains are being isolated with increasing frequency. From 1996 to 2000, 297 cases of urinary tract infection (UTI) due to QREC were observed in our hospital; 218 episodes (73.5%) were community acquired. The incidence of QREC UTIs increased steadily from 14.4% to 21.3% during 5 years when we compared the clinical characteristics of 60 QREC UTI with those of 80 quinolone-susceptible E. coli UTIs. Significant differences in susceptibility to various antibiotics were observed between the QREC and QSEC strains of E. coli. The multidrug resistance rate of QREC was much higher (38.3%) than those of quinolone susceptible isolates (18.8%). Prior fluoroquinolone use (p = 0.05), old age (p = 0.001), and a vegetative state (p = 0.03) were the independent risk factors for the acquisition of QREC UTI. The outcome of E. coli UTI is dependent on quinolone resistance. Thirty-day mortality was higher in QREC UTI patients, probably due to aggravation of underlying illness, but not quinolone resistance. On the basis of PFGE analysis, although some clustering was found in the hospital, genomic diversity was found among both the community and nosocomial strains. The increased frequency of QREC UTIs is thus not due to transmission of resistant strains but probably results from the selection of resistant strains from the endogenous flora of patients.     

Virulence profiles in uropathogenic Escherichia coli isolated from pregnant women and children with urinary tract abnormalities

Uropathogenic Escherichia coli is the leading etiologic agent of urinary tract infections, encompassing a highly heterogeneous group of strains. Although many putative urovirulence factors have been described, none of them appear in all uropathogenic E. coli strains, a fact that suggests that this group would be composed of different pathogenic subgroups. In this work, a study was performed on two collections of E. coli isolates proceeding from urine cultures from two groups of patients with urinary tract infection: pregnant women and children with urinary tract abnormalities. The isolates were analyzed for their virulence content and for their phylogeny by means of PCR determinations and of phenotypic assays. Associations among the virulence traits analyzed were searched for and this approach led to the identification of five urovirulence profiles. From a total of 230 isolates, 123 (53%) could be assigned to one of these profiles. A few loci appeared as markers of these profiles so that their presence allowed predicting the general virulence content of the strains. It is presumed that these conserved associations among the virulence functions would be devoted to ensure the coherence of the bacterial pathogenic strategy. In addition, three profiles appeared with significantly different frequencies depending on the host of origin of the isolates, indicating the existence of a correlation between the virulence content of the strains and their host specificity.     

Studies on beta-lactamases from Escherichia coli isolated from urinary tract infections

The beta-lactamase types present in 75 ampicillin and carbenicillin resistant E. coli were characterized using isoelectric focusing (IEF). The strains were isolated from patients with urinary tract infections from two geographically different areas of Denmark: 38 strains from Copenhagen and 37 strains from North Jutland. For 19 of the strains from Copenhagen and 18 of the strains from North Jutland, their beta-lactamase activity against nitrocefin and ampicillin, carbenicillin, benzylpenicillin, cloxacillin and cephaloridine was examined by a micro-iodometric and an UV-spectrophotometric assay. The strains from Copenhagen showed greater activity (p less than 0.001) against nitrocefin than the strains from North Jutland. The rate of hydrolysis of ampicillin was greater for the strains from Copenhagen than for the strains from North Jutland. Ninety-three per cent of the strains produced plasmid-mediated beta-lactamases, of which the most prevalent, TEM-1, was produced by 97 per cent of these strains, and OXA-1 by 3 per cent.     

Prevalence of adhesive genes among uropathogenic Escherichia coli strains isolated from patients with urinary tract infection in Mangalore

The study was carried out to detect the adhesive genes pap (pyelonephritis associated pili), sfa (S fimbrial adhesin) and afa (afimbrial adhesin) from Escherichia coli strains isolated in patients diagnosed with urinary tract infection (UTI). A total of 23% of the isolates were positive for pap, sfa and afa genes with a prevalence of 60.87% (14/23), 39.1% (9/23) and 39.1% (9/23), respectively. Prevalence of multiple adhesive genes was 8.7% (2/23) for pap and afa, 30.43% (7/23) for pap and sfa. Significant numbers of isolates were positive for Congo red binding (80%) and haemolysin production 60%. The prevalence of multiple adhesive genes indicate the potential to adhere and subsequently cause a systemic infection among UTI patients.     

Molecular epidemiologic approaches to urinary tract infection gene discovery in uropathogenic Escherichia coli

Urinary tract infection (UTI) is one of the most frequently acquired bacterial infections. The vast majority of UTIs are caused by a large, genetically heterogeneous group of Escherichia coli. This genetic diversity has hampered identification of UTI-related genes. A three-step experimental strategy was used to identify genes potentially involved in E. coli UTI transmission or virulence: epidemiologic pairing of a UTI-specific strain with a fecal control, differential cloning to isolated UTI strain-specific DNA, and epidemiologic screening to identify sequences among isolated DNAs that are associated with UTI. The 37 DNA sequences initially isolated were physically located all over the tester strain genome. Only two hybridized to the total DNA of the sequenced E. coli K-12 strain; eight sequences were present significantly more frequently in UTI isolates than in fecal isolates. Three of the eight sequences matched to genes for multidrug efflux proteins, usher proteins, and pathogenicity island insertion sites, respectively. Using population characteristics to direct gene discovery and evaluation is a productive strategy applicable to any system.     

Alpha-hemolytic strains of Escherichia coli isolated in urinary tract infections in small children]

The authors focused attention on testing of biological properties of alpha-haemolytic strains of E. coli in infections of the urinary pathways in children and on the antibody response against alpha-haemolysin. The strains belonged into serogroups 02, 04, 06, 018, 075, 0112. The majority of strains had fimrial adhesins, i.e. P fimbriae, or fimbriae type 1. Alpha-haemolysin one of the factors of UPEC virulence induces anti-alphahaemolysin formation in serum (ANTI AH). Titres of ANTI AH in sera of children with acute uroinfection were from 1:32 to 1:1024. Lower titres were recorded in infections of the lower urinary pathways higher ones in renal infections, whereby the dynamics of ANTI AH titres depend on the development of the disease.     

Virulence factors in Escherichia coli urinary tract infection.

Uropathogenic strains of Escherichia coli are characterized by the expression of distinctive bacterial properties, products, or structures referred to as virulence factors because they help the organism overcome host defenses and colonize or invade the urinary tract. Virulence factors of recognized importance in the pathogenesis of urinary tract infection (UTI) include adhesins (P fimbriae, certain other mannose-resistant adhesins, and type 1 fimbriae), the aerobactin system, hemolysin, K capsule, and resistance to serum killing. This review summarizes the virtual explosion of information regarding the epidemiology, biochemistry, mechanisms of action, and genetic basis of these urovirulence factors that has occurred in the past decade and identifies areas in need of further study. Virulence factor expression is more common among certain genetically related groups of E. coli which constitute virulent clones within the larger E. coli population. In general, the more virulence factors a strain expresses, the more severe an infection it is able to cause. Certain virulence factors specifically favor the development of pyelonephritis, others favor cystitis, and others favor asymptomatic bacteriuria. The currently defined virulence factors clearly contribute to the virulence of wild-type strains but are usually insufficient in themselves to transform an avirulent organism into a pathogen, demonstrating that other as-yet-undefined virulence properties await discovery. Virulence factor testing is a useful epidemiological and research tool but as yet has no defined clinical role. Immunological and biochemical anti-virulence factor interventions are effective in animal models of UTI and hold promise for the prevention of UTI in humans.     

Molecular characterisation of rough variants of Vibrio cholerae isolated from hospitalised patients with diarrhoea

Seven rough isolates of Vibrio cholerae isolated as the sole infecting agent from patients with cholera-like diarrhoea were examined for the presence of the regulatory element toxR and certain virulence-associated genes of the CTX genetic element and V. cholerae pathogenicity island (VPI). Multiplex PCR analysis with wb-specific genes of either O1 or O139 origin showed that six of the seven isolates produced an O1 wb-specific amplicon and the remaining isolate produced an O139-specific amplicon. Analysis of lipopolysaccharide profiles of smooth variants of V. cholerae revealed the presence of long repeated units of 'O' polysaccharide side chains but all the rough variants appeared to be devoid of the latter and possessed only core oligosaccharide. PCR amplification with primers specific to the ctxA, ctxB, tcpA, tagA, int, aldA, toxT, LJ, RJ and toxR genes revealed that six of the seven rough isolates were positive for these genes. One isolate was found to be negative for tagA and RJ, indicating the presence of an altered VPI. Each of these isolates showed media-dependent expression of cholera toxin (CT) and produced more toxin than the reference V. cholerae O1 El Tor strain VC20 or O139 strain SG24 under comparable conditions. Studies on the clonality of these isolates by the analysis of rRNA genes indicated their relatedness to strains of V. cholerae O1 El Tor or O139, isolated during the same time period.     

Molecular epidemiology of Staphylococcus saprophyticus isolated from women with uncomplicated community-acquired urinary tract infection

Staphylococcus saprophyticus is a common cause of urinary tract infections (UTIs) in women. Little is known about the molecular epidemiology of S. saprophyticus UTIs. In the current study, we compared 76 isolates of S. saprophyticus prospectively isolated from women with uncomplicated UTI participating in a randomized placebo-controlled treatment trial performed in northern Sweden from 1995 to 1997 with 50 strains obtained in 2006 from five different locations in northern Europe with pulsed-field gel electrophoresis (PFGE). The aim was to elucidate the molecular epidemiology of this uropathogenic species and to investigate whether specific clones are associated with UTI in women. A total of 47 different PFGE profiles were detected among the 126 analyzed isolates. Ten clusters consisting of 5 to 12 isolates each showing PFGE DNA similarity of >85% were identified. Several clusters of genetically highly related isolates were detected in the original trial as well as among isolates obtained during 2006 from different locations. In the original trial, clonal persistence was found among 16 of 21 (76%) patients examined in the placebo group at follow-up 8 to 10 days after inclusion, indicating a low spontaneous short-time bacteriological cure rate. We conclude that multiple clones of S. saprophyticus were causing lower UTIs in women. The result suggests that some human-pathogenic clones of S. saprophyticus are spread over large geographical distances and that such clones may persist over long periods of time.     

Attachment of Escherichia coli to urinary sediment epithelial cells from urinary tract infection-prone and healthy children.

Escherichia coli isolated from patients with recurrent urinary tract infections were tested for ability to attach to human urinary sediment epithelial cells in vitro. Higher mean capacity to bind bacteria was found for epithelial cells of the patient from whom the E. coli strain had been isolated than for epithelial cells from subjects without a history of urinary tract infection. The two populations were age matched. No relation was found between the age of the cell donor (0 to 15 years) and the capacity for E. coli attachement.     

Identification of urovirulence traits in Escherichia coli by comparison of urinary and rectal E. coli isolates from dogs with urinary tract infection

Spontaneously occurring urinary tract infection (UTI) in dogs was exploited as an experiment of nature to gain insights into UTI pathogenesis in humans. Concurrent urinary and rectal Escherichia coli isolates from 37 dogs with UTI were compared with respect to phylogenetic background, O antigens, and extended virulence genotype. In 54% of the UTI episodes, the dog's urinary and rectal isolates represented the same strain. Urinary isolates differed dramatically from rectal-only isolates in that they derived predominantly from E. coli phylogenetic group B2, expressed typical (human) UTI-associated O antigens, and possessed many virulence-associated genes, most notably pap elements (P fimbriae), papG (adhesin) allele III, sfa/foc and sfaS (S fimbriae), hly (hemolysin), fyuA (yersiniabactin), iroN (siderophore), and ompT (outer membrane protease T). The 20 urinary isolates that corresponded with the host's predominant rectal strain were no less virulent according to the markers analyzed than were the 17 urinary isolates that differed from the host's predominant rectal strain. These findings suggest that UTI pathogenesis is similar in dogs and humans, provide added support for the special-pathogenicity over the prevalence hypothesis of UTI pathogenesis, and identify numerous specific virulence-associated factors as significant correlates of urovirulence.     

Nosocomial urinary tract infection with a slowly growing, fastidious Escherichia coli.

Escherichia coli recovered from a nosocomially acquired urinary tract infection required 48 h of incubation on blood agar and did not grow on other routine clinical laboratory media. This bacterium dissociated readily into three colony types, all of which were confirmed as E. coli by DNA hybridization studies. Preliminary studies indicate a prolonged lag phase that could not be corrected by the addition of a variety of peptones and yeast extracts. Better growth was achieved by the addition of 10% horse serum.