Chloramphenicol reduces phage resistance evolution by suppressing bacterial cell surface mutants

Abstract

Bacteriophages infect Gram-negative bacteria by attaching to molecules present on the bacterial outer membrane, often lipopolysaccharides (LPS). Modification of LPS can lead to resistance to phage infection. In addition, LPS modifications can impact antibiotic susceptibility, allowing for phage-antibiotic synergism. The evolutionary mechanism(s) behind such synergistic interactions remain largely unclear. Here, we show that the presence of antibiotics can affect the evolution of resistance to phage infection, using phage {Phi}X174 and Escherichia coli C. We use a collection of 34 E. coli C LPS strains, each of which is resistant to {Phi}X174, and has either a "rough" or "deep rough" LPS phenotype. Growth of the bacterial strains with the deep rough phenotype is inhibited at low concentrations of chloramphenicol (and, to a much lesser degree, gentamicin). Treating E. coli C wildtype with {Phi}X174 and chloramphenicol eliminates the emergence of mutants with the deep rough phenotype, and thereby slows the evolution of resistance to phage infection. At slightly lower chloramphenicol concentrations, phage resistance rates are similar to those observed at high concentrations; yet, we show that the diversity of possible mutants is much larger than at higher chloramphenicol concentrations. These data suggest that specific antibiotic concentrations can lead to synergistic phage-antibiotic interactions that disappear at higher antibiotic concentrations. Overall, we show that the change in survival of various {Phi}X174-resistant E. coli C mutants in the presence of antibiotics can explain the observed phage-antibiotic synergism. Classification: Biological Sciences, Evolution