A comparison of phylogenetic group, virulence factors and antibiotic resistance in Russian and Norwegian isolates of Escherichia coli from urinary tract infection

Isolates of Escherichia coli from 31 Norwegian and 31 Russian females with significant bacteruria who presented with clinical signs of urinary tract infection (UTI) were tested for antimicrobial sensitivity, the presence of virulence genes, phylogroup distribution and clonal affinity. Twenty isolates, representing the full clonal diversity of a collection of 138 intestinal isolates of E. coli from healthy Norwegian females, served as a reference group. Russian UTI isolates belonged more often to phylogroup A and possessed fewer virulence genes than did Norwegian isolates. UTI isolates of E. coli were genetically heterogeneous and had a high degree of antimicrobial sensitivity.     

Antimicrobial susceptibility in Escherichia coli of human and avian origin—a comparison of wild-type distributions

In the present study, the antimicrobial susceptibilities of 97 Escherichia coli isolates from birds, and 100 clinical isolates from blood cultures, were determined by disk diffusion. The wild-type distributions were defined by the normalized resistance interpretation method. It is shown that the avian and clinical inhibition zone diameter distributions of wild-type E. coli are indistinguishable.     

Pathogenicity island markers in commensal and uropathogenic Escherichia coli isolates

Uropathogenic isolates of Escherichia coli (UPEC) contain blocks of DNA, termed pathogenicity islands (PAIs), that contribute to their virulence. Two multiplex PCR assays were developed to detect eight PAI markers among 50 commensal E. coli and 100 UPEC isolates. In total, 40% of commensal isolates and 93% of UPEC carried PAIs. Despite this difference, the distribution of various PAIs showed the same pattern in both groups, with the most prevalent being PAI IV(536) (38% commensal vs. 89% UPEC), followed by PAI I(CFT073) (26% vs. 73%), PAI II(CFT073) (14% vs. 46%), PAI II(J96) (8% vs. 34%), PAI I(536) (8% vs. 33%) and PAI II(536) (4% vs. 20%). PAI III(536) was detected only in UPEC (2%), while PAI I(J96) was not detected in any isolate. Although the mean number of PAIs per isolate was higher among UPEC (2.97) than in commensal (0.98) isolates, there were no statistical differences among group B2 E. coli from the two origins; however, commensal isolates from groups D and B1 appeared to be less virulent than pathogenic isolates. Regardless of their phylogenetic group, nearly all the commensal and UPEC isolates with the same number of PAIs had the same PAI combinations. Although group B2 E. coli are uncommon among commensal intestinal flora, they are highly virulent when present, suggesting that the intestinal flora may act as a reservoir for bacteria that can cause urinary tract infection.     

Molecular epidemiology of Escherichia coli causing neonatal meningitis

Escherichia coli is the second cause of neonatal meningitis which is a major cause of neonatal mortality and is associated with a high incidence of neurological sequelae. E. coli neonatal meningitis (ECNM) strains, as other extraintestinal pathogenic E. coli, mainly belong to the phylogenetic group B2 and to a lesser extent to group D, but are distributed in fewer clonal groups. One of these, the O18:K1:H7 clone is worldwide distributed meanwhile others such as O83:K1 and O45:K1 are restricted to some countries. Over the past few years, major progress has been made in the understanding of the pathophysiology of E. coli O18:K1:H7 neonatal meningitis. In particular, specific virulence factors have been identified and are known to be carried by ectochromosomal DNA in most cases. Molecular epidemiological studies, including characterization of virulence genotypes and phylogenetic analysis are important to lead to a comprehensive picture of the origins and spread of virulence factors within the population of ECNM strains. To date, all the known genetic determinants obtained in ECNM strains are not sufficient to explain their virulence in their globality and further studies on clonal groups different from the archetypal O18:K1:H7 clone are needed. These studies would serve to find common pathogenic mechanisms among different ECNM clonal groups that may be used as potential target for a worldwide efficacious prevention strategy.     

Association between mortality of Escherichia coli meningitis in young infants and non-virulent clonal groups of strains

To identify factors associated with Escherichia coli meningitis (ECM) mortality in infants aged <3 months, the clinical, biological and bacterial characteristics of isolates from 99 cases of ECM were compared, including the phylogenetic group, multilocus sequence type, O serogroup and sequence O type (a combination of sequence type complex (STc) and O serogroup) and virulence genotype. All 99 isolates were susceptible to the initial antimicrobial treatment. The mortality rate (14%) was not influenced by term or post-natal age. Hypotension or seizures were the sole clinical predictive factors for fatal outcome (p <0.01), and abnormal initial trans-fontanellar ultrasound was associated with death (p 0.03). Seventy-seven isolates belonged to the common sequence O types (STc29^O1, STc29^O18, STc29^O45, STc301^O7, STc304^O16, STc697^O83, STc700^O1) causing neonatal meningitis. None of the phylogenetic groups and none of the virulence determinants were distributed differently between survivors and non-survivors, except that the aerobactin gene ( iucC ) was less frequent in lethal isolates (94% vs. 71%, p 0.02). Isolates belonging to rare sequence O types were more likely to be lethal (OR 4.3, p 0.01), although they induced a lower level of bacteraemia than common sequence O types such as STc29^O18 and STc29^O45 in a neonatal rat model. These results suggest that unidentified human genetic risk-factors may be more important than strain virulence in predicting ECM mortality.     

Production by Escherichia coli isolates of siderophore and other virulence factors and their pathogenic role in a cutaneous infection model

Escherichia coli isolates from urinary tract infections (UTIs) (n = 124), extra-urinary sites (n = 37) and normal faecal samples (n = 51) were examined for the presence of virulence factors, including siderophores (aerobactin and enterobactin). The proportion of aerobactin producers was significantly higher in UTI (69.4%; p 0.001) and extra-urinary samples (70.3%; p 0.007) than in controls (41.2%), while the proportion of enterobactin producers was significantly lower in the UTI samples than in the controls (p 0.027). In a cutaneous infection model, aerobactin-positive E. coli showed more growth than non-aerobactin and non-enterobactin isolates, even when other virulence factors were identical.     

Phylogenetic groups and virulence factors of Escherichia coli strains causing pyelonephritis in children with and without urinary tract abnormalities

Escherichia coli isolates causing acute pyelonephritis in 93 children (25% with urinary tract abnormalities) were tested for nine virulence factors (papC, papGII, papGIII, sfa/foc, hlyC, cnf1, iucC, fyuA and iroN) and their phylogenetic groups were determined. Isolates lacking papGII were more frequent among patients with urinary tract abnormalities (58% vs. 10%, p 0.0003), as were non-virulent phylogenetic group A isolates (25% vs. 5%, p 0.043). Pyelonephritis caused by less virulent E. coli strains was more frequent among patients with significant urinary tract abnormalities. Further studies are required to determine whether screening for E. coli virulence factors may help to identify children warranting anatomical investigations.     

Analysis of quorum sensing-dependent virulence factor production and its relationship with antimicrobial susceptibility in Pseudomonas aeruginosa respiratory isolates

Pseudomonas aeruginosa is an opportunistic pathogen causing severe respiratory infections. The pathogenesis of these infections is multifactorial and the production of many virulence factors is regulated by quorum sensing (QS), a cell-to-cell communication mechanism. The two well defined QS systems in P. aeruginosa, the las and rhl systems, rely on N-acyl homoserine lactone signal molecules, also termed autoinducers. We assessed the activity of QS-dependent virulence factors (including elastase, alkaline protease, pyocyanin and biofilm production) in respiratory isolates of P. aeruginosa and their relationship with antimicrobial susceptibility. We identified sixteen isolates displaying impaired phenotypic activity; among them, eleven isolates were also defective in autoinducer production, and therefore considered QS-deficient. Six of the QS-deficient isolates failed to amplify one or more of the four QS regulatory genes (lasI, lasR, rhlI, rhlR) with PCR: one isolate was negative for rhlR, two isolates were negative for rhlI and rhlR and three isolates were negative for all four genes. The isolates that were negative for virulence factor production were generally less susceptible to the antimicrobials and statistically significant correlations were observed between the lack of elastase production and resistance to piperacillin and ceftazidime; between failure in alkaline protease production and resistance to tobramycin, piperacillin, piperacillin-tazobactam, cefepime, imipenem and ciprofloxacin; and between failure in pyocyanin production and resistance to amikacin, tobramycin, ceftazidime, ciprofloxacin and ofloxacin. The results obtained indicate that, despite the pivotal role of QS in the pathogenesis of P. aeruginosa respiratory infections, QS-deficient strains are still capable of causing infections and tend to be less susceptible to antimicrobials.     

新生儿脑膜炎大肠埃希菌 ompT 敲除株毒力分析

目的探讨新生儿脑膜炎大肠埃希菌ompT对菌株毒力的影响。方法对比野生致病株E44及ompT敲除株E44∶ΔompT两者体外黏附人脑微血管内皮细胞的能力;构建回复株,并考察ompT回补质粒pST及点突变质粒pST85能否恢复敲除株的黏附能力;考察ompT缺失对菌株定植乳鼠肠道能力的影响;考察 ompT 缺失对菌株穿越乳鼠血脑屏障的影响。结果相比野生株, E44∶ΔompT黏附人脑微血管内皮细胞能力明显降低;低拷贝质粒pST及点突变质粒pST85部分恢复了E44∶ΔompT的黏附能力;ompT缺失不影响菌株肠道定植能力;E44∶ΔompT可诱导与野生株相似水平的菌血症,但穿越乳鼠血脑屏障的能力显著降低。结论 ompT与菌体黏附脑微血管内皮细胞及穿越血脑屏障相关,OmpT蛋白水解酶活性在其中的作用有待进一步研究。     

Antimicrobial susceptibility trends among Escherichia coli and Shigella spp. isolated from rural Egyptian paediatric populations with diarrhoea between 1995 and 2000

Antimicrobial susceptibility testing was performed on 3,627 isolates of Escherichia coli and 180 isolates of Shigella spp. collected in rural locations from 875 Egyptian children with diarrhoea between 1995 and 2000. The cumulative rates of resistance for E. coli and Shigella spp. were high (respectively, 68.2% and 54.8% for ampicillin, 24.2% and 23.5% for ampicillin-sulbactam, 57.2% and 42.5% for trimethoprim-sulphamethoxazole, and 50.9% and 75.4% for tetracycline). Non-enterotoxigenic E. coli (NETEC) isolates had a consistently higher level of antimicrobial resistance than did enterotoxigenic E. coli (ETEC) isolates. Trend testing showed significant decreases in resistance to ampicillin, ampicillin-sulbactam and tetracycline among all E. coli isolates. Increasing rates of resistance were observed for trimethoprim-sulphamethoxazole in ETEC isolates and Shigella spp., but not in NETEC isolates. Low levels of resistance were observed for all other antimicrobial agents tested. Overall, high levels, but decreasing trends, of resistance to commonly used antimicrobial agents were detected among isolates of E. coli and Shigella spp. from children in rural Egypt.     

Antimicrobial resistances of extraintestinal pathogenic Escherichia coli isolates from swine in China

Antibiograms and relevant genotypes of porcine extraintestinal pathogenic Escherichia coli (ExPEC) isolates (n = 315) recovered between 2004 and 2007 in China were assessed. Among the 14 antimicrobials tested, the most prevalent resistance was to ampicillin, trimethoprim, sulfadimidine, tetracycline, neomycin, streptomycin, kanamycin, ciprofloxacin and ofloxacin (ranging from 81.9 to 100%). Forty-six multiresistant patterns were found. For each antimicrobial agent, ampicillin resistance was primarily mediated by bla_TEM, streptomycin resistance by strA and strB, kanamycin/neomycin resistance by aphA1, gentamicin resistance by aac(3)-IV, quinolones resistance by mutations in gyrA, tetracycline resistance by tet(A), tet(B) and tet(G), trimethoprim resistance by dfrA7, dfrA12 and dfrA13, and sulfadimidine resistance by sul1 and sul2. Both bla_TEM-1 and bla_CTX-M-14 were found in two ESBLs-producing isolates. Strains that harbored several genes that conferred resistance to the same antimicrobial agent were often significantly more multiresistant than others. Class 1 integrons were identified in 86 (27.3%) ExPEC isolates, which harbored dfrA14, aadA2, aadA22, dfrA17, aadA5, dfrA17-aadA2, dfrA1-aadA1, dfrA12-aadA2, dfrA17-aadA5 gene cassettes in five major different variable regions, conferring resistance to trimethoprim and aminoglycosides. These results provide novel insights into the epidemiological characteristics of porcine ExPEC strains in China, and suggest the need for the prudent use of antimicrobial agents in food animals.     

Phylogenetic group B2 Escherichia coli strains from the bowel microbiota of Pakistani infants carry few virulence genes and lack the capacity for long-term persistence

Escherichia coli strains of phylogenetic group B2 obtained from Western human hosts are enriched in virulence-associated genes and have a superior capacity to persist in the colonic microbiota. Here, E . coli strains from 22 infants born in Pakistan whose rectal flora was sampled regularly over the first 6 months of life were examined. B2 strains did not carry the virulence-associated genes sfaD/E , papC , neuB or hlyA more often than strains of other phylogenetic groups . B2 origin was not associated with persistence in the bowel microbiota. As compared with B2 strains from Swedish infants, Pakistani B2 strains carried significantly less often the virulence genes fimH (p 0.04), papC (p 0.02), papG class III (p 0.01), sfaD / E (p ≤0.0001), neuB (p ≤0.0001), and hlyA (p 0.005), and also the high-pathogenicity island (p ≤0.0001). A minority of Pakistani B2 strains belonged to recognized uropathogenic O-groups, which are common among 'Western' B2 strains. Thus, extra-intestinal pathogenicity may be the foremost characteristic of B2 strains colonizing Western populations.     

Attaching and effacing Escherichia coli isolates from Danish children: clinical significance and microbiological characteristics

This study describes the prevalence, clinical manifestations and microbiological characteristics of attaching and effacing Escherichia coli isolates, i.e., enteropathogenic E. coli (EPEC) belonging to the classical EPEC serotypes, non-EPEC attaching and effacing E. coli (A/EEC) and verocytotoxin-producing E. coli (VTEC), isolated in a case-control study of Danish children aged <5 years. Among 424 children with diarrhoea and 866 healthy controls, EPEC and VTEC were more prevalent in cases (2.4% and 2.6%, respectively) than in controls (0.7% and 0.7%, respectively). There was a high frequency of A/EEC isolates (n = 121), but these were equally prevalent in cases (11.3%) and controls (12.5%), and comprised a heterogeneous distribution of O:H serotypes. The intimin (eae) subtypes in A/EEC isolates showed an even distribution; the eae-gamma subtype predominated in classical EPEC cases. The virulence genes encoding the bundle-forming pilus (bfpA) and enteroaggregative heat-stable enterotoxin (astA) were rare among all isolates, and seemed to be of limited pathogenic importance in this population. Virulence characterisation of A/EEC isolates did not reveal any significant differences between cases and controls. Colonisation of children with A/EEC was associated with contact with sheep or goats (OR 2.2). The role of A/EEC, not being VTEC or belonging to the classical EPEC serotypes, requires further clarification, but serotyping is useful in discriminating between EPEC and A/EEC strains.     

Patient and bacterial determinants involved in symptomatic urinary tract infection caused by Escherichia coli with and without bacteraemia

Risk-factors for bacteraemia were determined in a case-control study of patients with Escherichia coli urinary tract infection. Cases were defined as patients with E. coli urinary source bacteraemia, and controls were chosen from among patients with E. coli urinary tract infection without bacteraemia. Patient characteristics were collected prospectively and the bacterial traits were determined. The phylogenetic background and virulence factors of E. coli isolates did not differ between cases and controls. In multivariate analysis, being female and having a urinary catheter were significantly less prevalent among patients with urinary source bacteraemia than among patients with uncomplicated urinary tract infection.     

Antibiotic resistance of Escherichia coli from community-acquired urinary tract infections in relation to demographic and clinical data

This prospective study determined the antibiotic susceptibility of 164 isolates of Escherichia coli from the urine of 164 patients (112 female, 52 male; mean age of 54.12 years) with community-acquired urinary tract infection (UTI). Half of the isolates were from uncomplicated UTI and half from complicated UTI (52 males and 34 females). Overall, 57.3% of isolates were resistant to ampicillin, 25% to co-trimoxazole, 20.1% to nalidixic acid, 14% to norfloxacin and ciprofloxacin, and 0% to fosfomycin and nitrofurantoin. Of the 82 isolates from complicated UTI, 16 (19.5%) were resistant to norfloxacin and ciprofloxacin, compared with seven (8.5%) from uncomplicated UTI (p 0.043). Isolates from patients aged >50 years were significantly more resistant than those from patients aged <50 years for nalidixic acid (p 0.007) and the fluoroquinolones tested (p 0.015). Resistance to fluoroquinolones was 25% (13/52) in males and 9% (10/112) in females (p 0.006). For patients with and without previous antimicrobial therapy, there was a significant difference only for resistance to nalidixic acid (p < 0.001) and the fluoroquinolones (p 0.011). There were adequate susceptibility rates to fosfomycin, nitrofurantoin and the fluoroquinolones for empirical use in the treatment of acute uncomplicated UTI. In order to interpret cumulative susceptibility data from the primary healthcare setting, it is necessary to take into account the type of UTI (uncomplicated vs. complicated), previous antimicrobial therapy, and the sex and age of each patient.     

Evolution of fluoroquinolone resistance among Escherichia coli urinary tract isolates from a French university hospital: application of the dynamic regression model

Escherichia coli urinary tract isolates were collected in 1997-2003 from Nimes University Hospital in order to investigate long-term trends in antibiotic resistance and to explore the relationship between antibiotic use and the emergence of resistance. Time-series analysis (ARIMA models) and dynamic regression models were used to investigate relationships between antibiotic use and resistance to ofloxacin and ciprofloxacin. Significant increases were seen in the frequency of ofloxacin (8.9 to 16.7%) and ciprofloxacin resistance (6.2 to 10.1%) (p < 0.001). Using multivariate dynamic regression analysis, it was found that an increased use of one defined daily dose (DDD)/1000 patient-days for ofloxacin, ciprofloxacin and norfloxacin induced average increases of 0.81%, 0.65% and 0.53% in E. coli ofloxacin resistance (p < 0.01), with average delays of 4, 4 and 6 months, respectively. An increase of 1 DDD/1000 patient-days of ciprofloxacin, ofloxacin and norfloxacin use induced increases of 0.73%, 0.82% and 0.63% in E. coli ciprofloxacin resistance (p < 0.01), with average delays of 4, 4 and 5 months, respectively. The use of nalidixic acid was not associated significantly with an increase in resistance to fluoroquinolones by multivariate analysis.     

Detection of virulence factors in α-haemolytic Escherichia coli strains isolated from various clinical materials

In total, 201 alpha-haemolytic Escherichia coli isolates from various clinical materials (urine samples and vaginal and rectal swabs) were examined by PCR for the presence of genes for the virulence factors alpha-haemolysin (hly), cytotoxic necrotising factor type 1 (cnf1), P-fimbriae (pap), S/F1C-fimbriae (sfa/foc), aerobactin (aer) and afimbrial adhesin (afaI). Among vaginal isolates, 96% were positive for cnf1, compared with 80% of urine strains (p 0.02) and 63% of rectal strains (p 0.0001). Similarly, sfa/foc-specific DNA sequences were found in 97% of vaginal isolates compared with 75% of rectal strains (p 0.004). The afa1 and aer genes were associated more with rectal alpha-haemolytic E. coli strains than with extra-intestinal isolates. The results suggested that CNF1 and/or S/F1C-fimbriae contribute to colonisation and persistence of alpha-haemolytic E. coli strains in the vaginal environment.     

Effect of bacterial virulence on IL-18 expression in the amnion infected with Escherichia coli

PROBLEM: The upregulation of inflammatory substances threatens pregnancy. Interleukin-18 (IL-18) is elevated in women who miscarried. The purpose of this study was to develop a pig model of chorioamnionitis to study the effect of bacterial virulence on IL-18 response in experimentally infected amnion. METHOD OF STUDY: A total of 20,000 colony-forming units of Escherichia coli (an enteropathogenic O55 strain, EPEC or O86 non-pathogenic strain) were administered into the amniotic cavity of pig fetuses at 70% of gestation for 10 hr. Fetal amniotic fluid samples were analyzed for IL-18 levels by enzyme-linked immunosorbent assay. The expression of IL-18 was studied also by immunohistochemistry on cryostat sections through amniotic membranes and pathological changes were observed by electron microscopy. RESULTS: Both E. coli strains propagated in amniotic fluids and reached similar counts. Only EPEC, however, caused a significant increase of IL-18 amniotic fluid levels (P < 0.001) and cytokine expression in the amniotic epithelium. CONCLUSIONS: The levels of IL-18 in infected amniotic fluids correlated with bacterial virulence and pathological changes in the amnion.     

Gentamicin susceptibility in Escherichia coli related to the genetic background: problems with breakpoints

In total, 120 Escherichia coli isolates positive for one of the gentamicin resistance (GEN(R)) genes aac(3)-II, aac(3)-IV or ant(2')-I were tested for gentamicin susceptibility by the agar dilution method. Isolates positive for aac(3)-IV or ant(2')-I had an MIC distribution of 8-64 mg/L, whereas isolates positive for aac(3)-II had MICs of 32 to >512 mg/L, suggesting a relationship between the distribution of MICs and the specific GEN(R) mechanism. The MIC distribution, regardless of the GEN(R) mechanism, was 8 - >512 mg/L, which supports the clinical breakpoint of MIC >4 mg/L suggested by EUCAST and questions the breakpoint recommended by the CLSI (> or =16 mg/L).