Bacteriophages with the ability to degrade uropathogenic Escherichia coli biofilms - PubMed (original) (raw)

Bacteriophages with the ability to degrade uropathogenic Escherichia coli biofilms

Andrew Chibeu et al. Viruses. 2012 Apr.

Abstract

Escherichia coli-associated urinary tract infections (UTIs) are among the most common bacterial infections in humans. UTIs are usually managed with antibiotic therapy, but over the years, antibiotic-resistant strains of uropathogenic E. coli (UPEC) have emerged. The formation of biofilms further complicates the treatment of these infections by making them resistant to killing by the host immune system as well as by antibiotics. This has encouraged research into therapy using bacteriophages (phages) as a supplement or substitute for antibiotics. In this study we characterized 253 UPEC in terms of their biofilm-forming capabilities, serotype, and antimicrobial resistance. Three phages were then isolated (vB_EcoP_ACG-C91, vB_EcoM_ACG-C40 and vB_EcoS_ACG-M12) which were able to lyse 80.5% of a subset (42) of the UPEC strains able to form biofilms. Correlation was established between phage sensitivity and specific serotypes of the UPEC strains. The phages' genome sequences were determined and resulted in classification of vB_EcoP_ACG-C91 as a SP6likevirus, vB_EcoM_ACG-C40 as a T4likevirus and vB_EcoS_ACG-M12 as T1likevirus. We assessed the ability of the three phages to eradicate the established biofilm of one of the UPEC strains used in the study. All phages significantly reduced the biofilm within 2-12 h of incubation.

Keywords: UPEC; bacteriophage; biofilms.

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Figures

Figure 1

Figure 1

Urinary tract infection (UTI) E. coli isolates positive for biofilm formation on 96-well microtiter plates. Isolates were considered as biofilm formers if the OD600 for the crystal violet-stained biofilm was equal to or greater than 3-fold the OD600 for a bacteria free medium. Data points represent an average of three independent experiments, each with 6 replicate wells for each isolate tested. Isolate CTF073 is a positive control for biofilm formation. Error bars indicate standard error of means.

Figure 2

Figure 2

Negative staining of phages ACG-C91 (a); ACG-M12 (b) and ACG-C40 (c) with 2% uranyl acetate or 2% phosphotungstate. Final magnification is × 297,000. Bars indicate 100 nm.

Figure 3

Figure 3

Phage disruption of established E. coli strain Can 91 biofilm. Biofilms grown in polystyrene microtiter plate wells for 48 h, were initially inoculated with 105, 107 and 109 pfu of phages (a) ACG-C40 and (b) ACG-C91 (c) ACG-M12. After 2, 4, 8 and 24 h of phage treatment at 37°C, average biofilm biomass in corresponding microtiter plate wells, were scored relatively to untreated control samples (100%) and represented on the Y-axes. Three independent experiments were performed, each starting from a different overnight culture and each with six repeats for each parameter combination. Error bars indicate standard error of means.

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