Applied Microbiology and Biotechnology

, Volume 85, Issue 5, pp 1533–1540

Intercrossing of phage genomes in a phage cocktail and stable coexistence with Escherichia coli O157:H7 in anaerobic continuous culture

Applied Genetics and Molecular Biotechnology

DOI: 10.1007/s00253-009-2230-2

Cite this article as:
Kunisaki, H. & Tanji, Y. Appl Microbiol Biotechnol (2010) 85: 1533. doi:10.1007/s00253-009-2230-2

Abstract

The emergence of phage-resistant cells is the most serious problem for realizing phage therapy and is observed frequently if only one phage strain is used against a particular bacterium. By contrast, using multiple phages (phage cocktail) can delay or control the appearance of phage-resistant cells. Anaerobic continuous culturing of Escherichia coli O157:H7 and a cocktail of EP16, PP17, and SP22 phages were conducted. Comparison of the restriction fragment length polymorphism (RFLP) pattern of each phage genome showed a pattern different from wild type. Furthermore, the RFLP pattern of mutant phages consisted of fragments of PP17 and SP22 genome, suggesting both phages had infected the same host simultaneously (superinfection) and exchanged genomic DNA. Through observation of the binding of SYBR Gold-stained mutant phage to individual phage-resistant cells (RC), we found that clonal RC cultures were heterogeneous in their ability to bind mutant phage. The ratio of susceptibility was a few percent, which suggested that a minority of the RC population was susceptible to phage, and this heterogeneity contributes to the stable coexistence of RC and chimeric phages. The ratio of susceptible cells did not change appreciably from bacterial generation to generation.

Keywords

BacteriophagePhage therapyPhage resistanceE. coli O157:H7

Copyright information

© Springer-Verlag 2009

Authors and Affiliations

  1. 1.Department of BioengineeringTokyo Institute of TechnologyYokohamaJapan