Long-term effects of single and combined introductions of antibiotics and bacteriophages on populations of Pseudomonas aeruginosa

With escalating resistance to antibiotics, there is an urgent need to develop alternative therapies against bacterial pathogens and pests. One of the most promising is the employment of bacteriophages (phages), which may be highly specific and evolve to counter antiphage resistance. Despite an increased understanding of how phages interact with bacteria, we know very little about how their interactions may be modified in antibiotic environments and, reciprocally, how phage may affect the evolution of antibiotic resistance. We experimentally evaluated the impacts of single and combined applications of antibiotics (different doses and different types) and phages on invitro evolving populations of the opportunistic pathogen Pseudomonas aeruginosa PAO1. We also assessed the effects of past treatments on bacterial virulence invivo, employing larvae of Galleria mellonella to survey the treatment consequences for the pathogen. We find a strong synergistic effect of combining antibiotics and phages on bacterial population density and in limiting their recovery rate. Our long-term study establishes that antibiotic dose is important, but that effects are relatively insensitive to antibiotic type. From an applied perspective, our results indicate that phages can contribute to managing antibiotic resistance levels, with limited consequences for the evolution of bacterial virulence.