Using bacterial population dynamics to count phages and their lysogens

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.

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Publication Details

Journal
Nature Communications
Published
2024-09-06
DOI
https://doi.org/10.1038/s41467-024-51913-6
Citations
31
Primary Topic
Bacteriophages and microbial interactions
Type
article
Field-Weighted Citation Impact
13.77

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article

Using bacterial population dynamics to count phages and their lysogens

Thu Vu Phuc Nguyen, Yuncong Geng, Ehsan Homaee, Ido Golding
31 citations
Nature Communications
Bacteriophages and microbial interactions
13.77
article

Using bacterial population dynamics to count phages and their lysogens

Thu Vu Phuc Nguyen, Yuncong Geng, Ehsan Homaee, Ido Golding
article en
31 citations

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.

Nature CommunicationsVol. 15(1)
University of Illinois Urbana-Champaign (US), Baylor College of Medicine (US), Urbana University (US), Princeton University (US)
National Science Foundation, Alfred P. Sloan Foundation, National Institutes of Health, National Institute of General Medical Sciences
Zero hunger
Openalex Percentile: Top 1%
Bacteriophages and microbial interactions
13.77
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