Natural Bred epsilon(2)-Phages Have an Improved Host Range and Virulence against Uropathogenic Escherichia coli over Their Ancestor Phages

Alternative treatments for Escherichia coli infections are urgently needed, and phage therapy is a promising option where antibiotics fail, especially for urinary tract infections (UTI). We used wastewater-isolated phages to test their lytic activity against a panel of 47 E. coli strains reflecting the diversity of strains found in UTI, including sequence type 131, 73 and 69. The plaquing host range (PHR) was between 13 and 63%. In contrast, the kinetic host range (KHR), describing the percentage of strains for which growth in suspension was suppressed for 24 h, was between 0% and 19%, substantially lower than the PHR. To improve the phage host range and their efficacy, we bred the phages by mixing and propagating cocktails on a subset of E. coli strains. The bred phages, which we termed evolution-squared epsilon(2)-phages, of a mixture of Myoviridae have KHRs up to 23% broader compared to their ancestors. Furthermore, using constant phage concentrations, Myoviridae epsilon(2)-phages suppressed the growth of higher bacterial inocula than their ancestors did. Thus, the epsilon(2)-phages were more virulent compared to their ancestors. Analysis of the genetic sequences of the epsilon(2)-phages with the broadest host range reveals that they are mosaic intercrossings of 2-3 ancestor phages. The recombination sites are distributed over the whole length of the genome. All epsilon(2)-phages are devoid of genes conferring lysogeny, antibiotic resistance, or virulence. Overall, this study shows that epsilon(2)-phages are remarkably more suitable than the wild-type phages for phage therapy.