Development of a Bacteriophage Cocktail to Constrain the Emergence of Phage-Resistant Pseudomonas aeruginosa

Abstract

With the emergence of multidrug-resistant and extensively drug-resistant bacterial pathogens, phage therapy and other alternative or additional therapeutic modalities are receiving resurgent attention. One of the major obstacles in developing effective phage therapies is the evolution of phage resistance in the bacterial host. When Pseudomonas aeruginosa was infected with a phage that uses O-antigen as receptor, phage resistances typically achieved through changing or loss of O-antigen structure. In this study, we showed that dsRNA phage phiYY uses core lipopolysaccharide as receptor and therefore efficiently kills the O-antigen deletion mutants. Furthermore, by phage training, we obtained PaoP5-m1, a derivative of dsDNA phage PaoP5, which is able to infect mutants with truncated O-antigen. We then generated a cocktail by mixing phiYY and PaoP5-m1 with additional three wide host range P. aeruginosa phages. The phage cocktail was effective against a diverse selection of clinical isolates of P. aeruginosa, and in the short-term constrained the appearance of the phage-resistant mutants that had beleaguered the effectiveness of single phage. Resistance to the 5phage cocktail emerges after several days, and requires mutations in both wzy and migA Thus, this study provides an alternative strategy for designing phage cocktail and phage therapy.

Coevolutionary phage training leads to greater bacterial suppression and delays the evolution of phage resistance

“…Although resistance to phages is as or more common than to antibiotics, advantages of using phages as therapeutics have been proposed time and again (7,8,(19)(20)(21)(22). Notably, unlike antibiotics, phages are biological entities that evolve.…”

Transcriptomic Analysis Reveals the Dependency of Pseudomonas aeruginosa Genes for Double-Stranded RNA Bacteriophage phiYY Infection Cycle

“…Phage phiYY is effective in infecting the rough strains and has been included in a phage cocktail to limit the emergence of phage-resistance mutant ( Yang et al., 2020 ). PAO1r_8 is a phage-resistant mutant selected after dsDNA phage PaoP5 infection ( Shen et al., 2018 ).…”

An in‐vitro study on a novel six‐phage cocktail against multi‐drug resistant‐ESBL Shigella in aquatic environment

“…Besides that, not only the diversity of phages in the cocktail significantly lowers possible development of phage-resistant host but also even leading to severe, fast and stable reduction of pathogens. In fact, the more phages types that target particular host, the more significantly constrained mutants should take place for generating a new phage-resistant bacteria (Yang et al 2020) According to the results, this phage cocktail could be a worthy alternative for removing multi-drug-resistant ESBL-positive Shigella contamination or preventing contamination of water resources.…”