Control of residual phage in the evaluation of phage-based food safety applications

Bacteriophage ("Phage") products are gaining interest in controlling foodborne pathogens as they are natural, specific, and can replicate at the site of contamination. One challenge in determining the efficacy of phage biocontrol is accounting for residual phages that may impact the recovery and the enumeration of surviving bacteria downstream from the treatment on food surface (FS) or food contact surface (FCS). Typically, the efficacy of a phage formulation is tested by applying it to a FS or FCS that has been pre-inoculated with the target pathogen and incubating the treatment for a set period of time. The food sample is transferred into a liquid buffer and stomached to release the surviving bacteria for their enumeration. During these final steps, there is a potential for residual phage to interact with bacterial survivors, which could affect the calculated efficacy of the phage. Limited studies demonstrated that bacterial reductions in these experiments occur specifically during treatment and not during sample recovery. This review highlights the importance of including appropriate controls to determine if residual phages are impacting bacterial recovery and the methods used to mitigate those impacts, which involves either neutralizing residual phages or separating residual phages from the surviving bacteria.