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BackgroundCrohn's disease (CD) is a high morbidity chronic inflammatory disorder of unknown aetiology. Adherent-invasive Escherichia coli (AIEC) has been recently implicated in the origin and perpetuation of CD. Because bacterial biofilms in the gut mucosa are suspected to play a role in CD and biofilm formation is a feature of certain pathogenic E. coli strains, we compared the biofilm formation capacity of 27 AIEC and 38 non-AIEC strains isolated from the intestinal mucosa. Biofilm formation capacity was then contrasted with the AIEC phenotype, the serotype, the phylotype, and the presence of virulence genes.ResultsSpecific biofilm formation (SBF) indices were higher amongst AIEC than non-AIEC strains (P = 0.012). In addition, 65.4% of moderate to strong biofilms producers were AIEC, whereas 74.4% of weak biofilm producers were non-AIEC (P = 0.002). These data indicate that AIEC strains were more efficient biofilm producers than non-AIEC strains. Moreover, adhesion (P = 0.009) and invasion (P = 0.003) indices correlated positively with higher SBF indices. Additionally, motility (100%, P < 0.001), H1 type flagellin (53.8%, P < 0.001), serogroups O83 (19.2%, P = 0.008) and O22 (26.9%, P = 0.001), the presence of virulence genes such as sfa/focDE (38.5%, P = 0.003) and ibeA (26.9%, P = 0.017), and B2 phylotype (80.8%, P < 0.001) were frequent characteristics amongst biofilm producers.ConclusionThe principal contribution of the present work is the finding that biofilm formation capacity is a novel, complementary pathogenic feature of the recently described AIEC pathovar. Characterization of AIEC specific genetic determinants, and the regulatory pathways, involved in biofilm formation will likely bring new insights into AIEC pathogenesis.

In four patients with alkaline-encrusted cystitis, Corynebacterium group D2 was isolated from consecutive urine cultures and stones. Encrusted cystitis occurred in bladders harboring inflammatory or tumorous lesions in patients with chronic or recurrent urinary tract infections appearing after surgery or instrumentation. The urease activity of Corynebacterium group D2 and the neutralization of this enzyme by acetohydroxamic acid are shown. Clinical improvement, disappearance of struvite crystals, and decrease of the urine pH were obtained when these bacteria were eliminated from urine samples. Corynebacterium group D2 strains were highly resistant to many antimicrobial agents but were highly susceptible to norfloxacin and vancomycin when tested at two pHs (7.4 and 8.5).

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The minimum dosage of antibiotics that reduced mortality in bacteraemic rats inoculated with two different Escherichia coli isolates was determined in an attempt to study the therapeutic importance of the inoculum effect. Low mortality rates (0-5%) at 48 h were obtained when antibiotics with minimal or no inoculum effect (ampicillin, cefuroxime, cefoxitin and gentamicin) were administered to yield serum levels 5 to 14 times the MIC, while antibiotics with a pronounced inoculum effect (piperacillin, cefotaxime and aztreonam) had to be administered to yield serum levels 57 to more than 1000 times the MIC determined with a standard (low) inoculum. All of the antibiotics with inoculum effect studied here are administered empirically in clinical practice at a higher dose than the microbiological and pharmacokinetic data would indicate (in order to reach peak serum concentrations exceeding the MICs of the pathogens by 4-10 times). Our experiment suggests that such high and empirical doses of antibiotics with inoculum effect may be justified.

This work demonstrates that C. urealyticum produces biofilms on polystyrene plates and biofilm-associated organisms are much less susceptible to the bactericidal effect of the antibiotics; and the exposure of C. urealyticum to erythromycin may favour resistance selection. Overall, these results may explain the difficulties for bacterial eradication in chronic infections caused by C. urealyticum.

The susceptibility in vitro of 28 Corynebacterium group D2 strains, mainly isolated from urine, to fifteen antimicrobial agents and acetohydroxamic acid (AHA) was determined at two pH values. The bactericidal activity of four antimicrobials and AHA was studied in three reference strains in broth at two pHs and with two inoculum sizes. The activity of norfloxacin and AHA, against one selected strain, in broth and human urine, was also determined. Vancomycin, ofloxacin and norfloxacin were the most active agents tested. Norfloxacin acted bactericidally in broth and in human urine but was not synergic with AHA.

The in vitro susceptibility of 30 Corynebacterium group D2 strains to nine antimicrobial agents was determined. Vancomycin and norfloxacin were the most active agents tested. All strains were resistant to ampicillin and cephalothin, all except one were resistant to gentamicin, and the activity of erythromycin, novobiocin, tetracycline, and rifampin varied.