Abstract
Chronic wounds affect thousands of people worldwide, causing pain and discomfort to patients and represent significant economical burdens to health care systems. The treatment of chronic wounds is very difficult and complex, particularly when wounds are colonized by bacterial biofilms which are highly tolerant to antibiotics. Enterococcus faecium and Enterococcus faecalis are within the most frequent bacteria present in chronic wounds. Bacteriophages (phages) have been proposed as an efficient and alternative against antibiotic-resistant infections, as those found in chronic wounds. We have isolated and characterized two novel enterococci phages, the siphovirus vB_EfaS-Zip (Zip) and the podovirus vB_EfaP-Max (Max) to be applied during wound treatment. Both phages demonstrated lytic behavior against E. faecalis and E. faecium. Genome analysis of both phages suggests the absence of genes associated with lysogeny. A phage cocktail containing both phages was tested against biofilms formed in wound simulated conditions at a multiplicity of infection of 1.0 and a 2.5 log CFU.mL−1 reduction in the bacterial load after at 3 h of treatment was observed. Phages were also tested in epithelial cells colonized by these bacterial species and a 3 log CFU.mL−1 reduction was observed using both phages. The high efficacy of these new isolated phages against multi-species biofilms, their stability at different temperatures and pH ranges, short latent periods and non-cytotoxicity to epithelial cells suggest their therapeutic use to control infectious biofilms present in chronic wounds.
Highlights
Chronic wounds affect thousands of people worldwide, causing pain and discomfort to patients and represent significant economical burdens to health care systems
World Health Organization (WHO) encourages the scientific community and pharmaceutical industries to focus on the development of new antimicrobials to combat antibiotic resistant pathogens[14]
According to the morphological evaluation[20], E. faecalis phage Max belongs to the Siphoviridae family, having a non-contractile tail with 220 nm in length and 12 nm in width and a capsid with 58 nm in diameter (Fig. 1A)
Summary
Chronic wounds affect thousands of people worldwide, causing pain and discomfort to patients and represent significant economical burdens to health care systems. After three hours of infection, E. faecalis phage Max reduced the number of viable cells in about 2 log CFU.mL−1 (p < 0.05) (Fig. 4A). Concerning E. faecium phage Zip, it caused a reduction of approximately 1.5 log CFU.mL−1 between 3 and 6 h of infection (Fig. 4B).
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