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Sequential Phage Delivery Can Outperform Cocktails by Delaying Cross-Resistance Evolution.

Elizabeth C Stuart1, Justin R Meyer1

  • 1Department of Ecology, Behavior and Evolution, University of California San Diego, La Jolla, CA 92093, USA.

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Summary

Sequential bacteriophage therapy, using T2 followed by λtrn phages, proved more effective than simultaneous cocktails against Escherichia coli. This strategy better controlled bacterial evolution and resistance, optimizing phage therapy outcomes.

Keywords:
Escherichia colibacteriophage therapycross-resistanceexperimental evolutionphage cocktailsresistance evolutionsequential delivery

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Area of Science:

  • Microbiology
  • Evolutionary Biology
  • Biotechnology

Background:

  • Antimicrobial resistance necessitates novel therapeutic strategies.
  • Bacteriophage therapy is a promising alternative, but bacterial evolution often limits efficacy.
  • The optimal administration strategy for combination phage therapy remains unclear.

Purpose of the Study:

  • To compare the efficacy of simultaneous versus sequential administration of two bacteriophages (λtrn and T2) against Escherichia coli.
  • To investigate how delivery strategy influences bacterial resistance evolution during phage therapy.
  • To identify genetic mechanisms underlying phage resistance and cross-resistance.

Main Methods:

  • Experimental comparison of simultaneous and sequential phage administration in laboratory conditions.
  • Phenotypic assays to assess bacterial suppression and resistance development.
  • Whole genome sequencing to identify genetic mutations conferring resistance.

Main Results:

  • Sequential T2-first phage delivery achieved superior and more sustained bacterial suppression compared to simultaneous cocktails.
  • Treatment outcomes were highly dependent on phage administration order and timing.
  • Distinct genetic mutations, including in envZ, were identified as drivers of phage cross-resistance.

Conclusions:

  • The temporal ordering of phage administration significantly impacts therapeutic efficacy.
  • Sequential phage delivery can be a powerful strategy to manage bacterial evolution and overcome resistance.
  • Evolutionary steering principles can optimize the design of combination phage therapies.