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Validating Flow Cytometry as a Method for Quantifying Bdellovibrio Predatory Bacteria and Its Prey for Microbial
Ayo Ogundero1, Marta Vignola1, Stephanie Connelly1
1Infrastructure and Environment, School of Engineering, University of Glasgowgrid.8756.c, Glasgow, UK.
Microbiology Spectrum
|February 24, 2022
Summary
Flow cytometry (FCM) offers a rapid and accurate method for counting predatory bacteria, Bdellovibrio bacteriovorus. This validated protocol aids ecological studies of Bdellovibrio predation dynamics, potentially leading to new biofilm control strategies.
Area of Science:
- Microbiology
- Biotechnology
- Analytical Chemistry
Background:
- Antibiotic resistance and bacterial biofilms present growing global challenges.
- Predatory bacteria, like Bdellovibrio bacteriovorus, are explored as biological alternatives to antibiotics.
- Accurate enumeration methods are crucial for ecological studies of predator-prey dynamics.
Purpose of the Study:
- To develop and validate a flow cytometry (FCM) protocol for enumerating Bdellovibrio bacteriovorus.
- To compare FCM's accuracy and speed against traditional methods like optical density, PFU assay, and qPCR.
- To assess FCM's utility in distinguishing mixed populations of Bdellovibrio and prey bacteria.
Main Methods:
- Protocol development for Bdellovibrio cell enumeration using flow cytometry (FCM).
- Comparative analysis of FCM with optical density (OD), plaque-forming unit (PFU) assay, and quantitative PCR (qPCR).
- Validation of FCM by counting mixed populations of Bdellovibrio and Pseudomonas.
Main Results:
- Strong correlations were observed between FCM and PFU (ρ = 0.923) and FCM and qPCR (ρ = 0.987).
- A weaker correlation was found between FCM and optical density (ρ = 0.784).
- FCM successfully distinguished and counted mixed populations of Bdellovibrio and Pseudomonas.
Conclusions:
- Flow cytometry provides a rapid, accurate, and reproducible method for enumerating Bdellovibrio bacteriovorus.
- FCM is a suitable alternative to traditional methods, especially for high-throughput analysis of predator-prey interactions.
- Validated FCM protocols can advance research into Bdellovibrio applications for controlling pathogenic bacteria and biofilms.
