Validating flow cytometry as a method for quantifying Bdellovibrio predatory bacteria and Its prey for microbial ecology

Ogundero, A., Vignola, M. , Connelly, S. and Sloan, W. T. (2022) Validating flow cytometry as a method for quantifying Bdellovibrio predatory bacteria and Its prey for microbial ecology. Microbiology Spectrum, 10(1), e01033-21. (doi: 10.1128/spectrum.01033-21) (PMID:35196816) (PMCID:PMC8865432)

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Abstract

Bdellovibrio bacteriovorus is a predatory, Gram-negative bacteria that feeds on many pathogenic bacteria and has been investigated as a possible solution for mitigating biofilms in different fields. The application depends on more fundamental ecological studies into the dynamics between Bdellovibrio and their prey. To do so requires an accurate, reliable, and, preferably rapid, way of enumerating the cells. Flow cytometry (FCM) is potentially a rapid, accurate, and inexpensive tool for this, but it has yet to be validated in the enumeration of Bdellovibrio. In this study, we developed a protocol to measure the number of Bdellovibrio in samples of various densities using FCM and compared the results with those of other methods: optical density (OD), PFU assay (PFU), and quantitative PCR (qPCR). We observed a strong correlation between values obtained using FCM and PFU (ρ = 0.923) and FCM and qPCR (ρ = 0.987). Compared to optical density there was a much weaker correlation (ρ = 0.784), which was to be expected given the well-documented uncertainty in converting optical density (OD) to cell numbers. The FCM protocol was further validated by demonstrating its ability to distinguish and count mixed populations of Bdellovibrio and the prey Pseudomonas. Thus, the accuracy of FCM as well as its speed and reproducibility make it a suitable alternative for measuring Bdellovibrio cell numbers, especially where many samples are required to capture the dynamics of predator-prey interactions.

Item Type:Articles
Additional Information:This work was supported by Scottish Water, IBioIC, and the Scottish Funding Council (SFC) and by Engineering and Physical Research Council, UK grants (EP/K038885/1 & EP/ V030515/1). M.V. is supported by the Royal Academy of Engineering under the Research Fellowship scheme (RF\201819\18\198).
Keywords:Infectious Diseases, Cell Biology, Microbiology (medical), Genetics, General Immunology and Microbiology, Ecology, Physiology
Status:Published
Refereed:Yes
Glasgow Author(s) Enlighten ID:Connelly, Dr Stephanie and Ogundero, Mr Ayo and Vignola, Dr Marta and Sloan, Professor William
Authors: Ogundero, A., Vignola, M., Connelly, S., and Sloan, W. T.
College/School:College of Science and Engineering > School of Engineering > Infrastructure and Environment
Journal Name:Microbiology Spectrum
Publisher:American Society for Microbiology
ISSN:2165-0497
ISSN (Online):2165-0497
Published Online:23 February 2022
Copyright Holders:Copyright © 2022 Ogundero et al
First Published:First published in Microbiology Spectrum 10(1): e01033-21
Publisher Policy:Reproduced under a Creative Commons License

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Project CodeAward NoProject NamePrincipal InvestigatorFunder's NameFunder RefLead Dept
190641Synthetic Biology applications to Water Supply and RemediationSteven BeaumontEngineering and Physical Sciences Research Council (EPSRC)EP/K038885/1Research and Innovation Services
309846Decentralised water technologiesWilliam SloanEngineering and Physical Sciences Research Council (EPSRC)EP/V030515/1ENG - Infrastructure & Environment
305420Eco-Engineered Biofilters For Sustainable Removal Of Pesticides In Drinking WaterMarta VignolaRoyal Academy of Engineering (RAE)RF\201819\18\198ENG - Infrastructure & Environment