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Using bacterial population dynamics to count phages and their lysogens
Yuncong Geng1,2, Thu Vu Phuc Nguyen1,3,4, Ehsan Homaee1,2
1Department of Physics, University of Illinois Urbana-Champaign, Urbana, IL, 61801, USA.
Nature Communications
|September 6, 2024
Summary
This study introduces a high-throughput microplate reader assay to quantify bacteriophages and their lysogens. The method uses optical density to measure phage concentration and host lysogenization rates, offering high sensitivity and efficiency.
Area of Science:
- Microbiology
- Virology
- Biophysics
Background:
- Traditional bacteriophage and lysogen counting methods are inefficient and disruptive.
- There is a need for high-throughput, sensitive, and non-perturbative methods for quantifying viral populations and host-pathogen interactions.
Purpose of the Study:
- To develop and validate a microplate reader-based assay for high-throughput bacteriophage quantification.
- To enable the measurement of host lysogenization rates.
- To investigate the influence of host physiology on viral developmental programs.
Main Methods:
- Utilizing a microplate reader to measure the optical density (OD) dynamics of phage-infected bacterial cultures.
- Correlating the OD at lysis with initial phage concentration across a wide dynamic range.
- Employing mathematical modeling to infer phage growth parameters (growth rate, latent period, burst size).
- Incorporating antibiotic selection to quantify host lysogenization rates.
Main Results:
- The OD at which cultures lyse exhibits a linear relationship with the logarithm of the initial phage concentration, enabling quantification over nine orders of magnitude.
- The assay demonstrates single-phage sensitivity.
- Phage growth rate was successfully inferred and shown to be dependent on encounter rate, latent period, and burst size.
- Antibiotic selection allowed for the measurement of host lysogenization rates.
Conclusions:
- The developed microplate assay is a sensitive, high-throughput method for quantifying bacteriophages and their lysogens.
- Host physiology, specifically growth rate, impacts bacteriophage lytic growth and lysogeny propensity, as demonstrated by decreased lytic growth and increased lysogeny in slower-growing E. coli.
- This method provides a powerful tool for studying phage-host interactions and viral dynamics.
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