Development of a broad-host-range phage cocktail for biocontrol

The aims of this study were to investigate the incidence of different resistance mechanisms to phage K in a bank of Irish Staph aureus hospital strains; and to develop a broad host-range phage cocktail with enhanced lytic activity against those strains which were previously phage resistant. A bank of 180 Staph aureus strains, which included all the sequence types currently in existence in Ireland, were tested for sensitivity to phage K. Twenty nine strains were identified, which did not permit plaque formation. The phage resistance systems in the 29 strain were investigated and it was found that restriction modification (r-m) was evident in 24, an adsorption inhibition mechanism was evident in three, while two were resistant by an unidentified mechanism. Seventeen modified derivatives of phage K were developed which could circumvent all the r-m systems. Nevertheless, six of the modified phage were considered superior in terms of their individual host ranges. These six were pooled as a cocktail with phage K, which then lysed 24 of the 29 resistant strains (97.2% of the entire staphylococcal bank). In conclusion, phage resistant systems affecting phage K are common in Staph. aureus but it is possible to significantly broaden the host-range of this phage for biocontrol applications.