Using bacterial population dynamics to count phages and their lysogens

Affiliations
  • 1Department of Physics, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA.
  • 2Center for Biophysics and Quantitative Biology, University of Illinois Urbana-Champaign, Urbana, IL 61801, USA.
  • 3Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA.
  • 4Department of Molecular Biology, Princeton University, Princeton, NJ 08544, USA.
  • 5Department of Physics, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA. igolding@illinois.edu.
  • 6Center for Biophysics and Quantitative Biology, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA. igolding@illinois.edu.
  • 7Verna and Marrs McLean Department of Biochemistry and Molecular Biology, Baylor College of Medicine, Houston, TX, 77030, USA. igolding@illinois.edu.
  • 8Department of Microbiology, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA. igolding@illinois.edu.

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Abstract

Traditional assays for counting bacteriophages and their lysogens are labor-intensive and perturbative to the host cells. Here, we present a high-throughput infection method in a microplate reader, where the growth dynamics of the infected culture is measured using the optical density (OD). We find that the OD at which the culture lyses scales linearly with the logarithm of the initial phage concentration, providing a way of measuring phage numbers over nine orders of magnitude and down to single-phage sensitivity. Interpreting the measured dynamics using a mathematical model allows us to infer the phage growth rate, which is a function of the phage-cell encounter rate, latent period, and burst size. Adding antibiotic selection provides the ability to measure the rate of host lysogenization. Using this method, we found that when E. coli growth slows down, the lytic growth rate of lambda phages decreases, and the propensity for lysogeny increases, demonstrating how host physiology influences the viral developmental program.