Efficacy of three lytic bacteriophages for eradicating biofilms of multidrug-resistant Proteus mirabilis

Abstract

Proteus mirabilis is one of the most frequent causes of catheter-associated urinary tract infections (CAUTIs) owing to its capability to colonize and develop crystalline multidrug-resistant (MDR) biofilms. Here, we report the isolation and partial characterization of three novel bacteriophages, vB_PmiM-ES1a, vB_PmiM-ES1b, and vB_PmiM-ES1c, which were active against the planktonic form and biofilms of the MDR P. mirabilis strain ES01, isolated from CAUTIs in Egypt. The antibiotic susceptibility profile of the P. mirabilis isolates showed resistance to most of the antibiotics tested. The isolated phages were identified morphologically using TEM, and each appeared to have myovirus-like morphology. The three phages displayed strong lytic activity and a narrow host range, and they were stable at different ranges of temperatures and pH values. One-step growth kinetics showed a lysis time of 180 min with a burst size of 99.6, 95, and 86 PFU/cell for phage vB_PmiM-ES1a, vB_PmiM-ES1b, and vB_PmiM-ES1c, respectively. The three phages exhibited different digestion patterns using different restriction enzymes. The genome size was estimated to be 59.39 kb, 62.19 kb, and 52.07 kb for phage vB_PmiM-ES1a, vB_PmiM-ES1b, and vB_PmiM-ES1c, respectively. A phage cocktail including the three phages showed a potential ability to reduce and eradicate a biofilm formed by the MDR Proteus mirabilis EG-ES1. Accordingly, a phage cocktail of vB_PmiM-ES1a, vB_PmiM-ES1b, and vB_PmiM-ES1c is considered a promising candidate for use as a biocontrol agent against MDR Proteus mirabilis bacteria.