Identification of novel bacteriophages with therapeutic potential targeting Enterococcus faecalis

The Gram-positive opportunistic pathogen Enterococcus faecalis is frequently responsible for nosocomial infections in humans and represents one of the most common bacteria isolated from recalcitrant endodontic (root canal) infections. E. faecalis is intrinsically resistant to several antibiotics routinely used in clinical settings (such as cephalosporins and aminoglycosides) and can acquire resistance to vancomycin (vancomycin resistant enterococci, VRE). The resistance of E. faecalis to several classes of antibiotics and its capacity to form biofilms cause serious therapeutic problems. In this paper, we report the isolation of several bacteriophages that target E. faecalis strains isolated from the oral cavity of patients suffering root-canal infections. All phages isolated were Siphoviridae with similar tail lengths (200-250 nm) and icosahedral heads. The genome sequences of three isolated phages were highly conserved with the exception of predicted tail protein genes that diverge in sequence, potentially reflecting host range. The properties of the phage with the broadest host-range (SHEF2), was further characterised. We showed that this phage requires interaction with components of the major and variant region Enterococcal polysaccharide antigen (Epa) to engage in lytic infection. Finally, we explored the therapeutic potential of this phage and showed that it can eradicate E. faecalis biofilms formed in vitro on a standard polystyrene surface but also on a cross-sectional tooth-slice model of endodontic infection. We also show that SHEF2 cleared a lethal infection of zebrafish when applied in the circulation. We therefore propose that the phage described in this study could be used to treat a broad range of antibiotic resistant E. faecalis infections.