Using bacterial population dynamics to count phages and their lysogens
Yuncong Geng, Thu Vu Phuc Nguyen, Ehsan Homaee, Ido Golding
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. Traditional assays for counting bacteriophages and their lysogens are labor-intensive and perturbative to the host cells. Here, the authors present a high-throughput method that can be used to estimate the number of phages in a sample, by measuring growth dynamics of the infected culture in a microplate reader.