Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry

The use of flow cytometry to enumerate microorganisms is gaining traction over the traditional plate count technique on the basis of superior accuracy, precision and time-to-result. Here, we assessed the suitability of live/dead flow cytometry for the enumeration of mixed populations of probiotic ba...

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Bibliographic Details
Published in:BMC microbiology 2023-02, Vol.23 (1), p.48-48, Article 48
Main Authors: Tracey, Harry, Coates, Niall, Hulme, Eleri, John, Daniel, Michael, Daryn Robert, Plummer, Susan Frances
Format: Article
Language:English
Subjects:
Acids
Bacteria
Bifidobacterium bifidum
Dyes
Enumeration
Flow
Flow Cytometry
Lactobacillus acidophilus
Measurement
Microbial populations
Microorganisms
Mixtures
Morphology
Multistrain
Organisms
Plate count
Populations
Probiotic
Probiotics
Sensors
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Summary:The use of flow cytometry to enumerate microorganisms is gaining traction over the traditional plate count technique on the basis of superior accuracy, precision and time-to-result. Here, we assessed the suitability of live/dead flow cytometry for the enumeration of mixed populations of probiotic bacteria (L. acidophilus, L. paracasei, L. plantarum, L. salivarius, B. lactis and B. bifidum) whilst comparing outcomes with plate counting. Using a novel gating strategy designed specifically for the enumeration of mixed populations, the application of flow cytometry resulted in the detection of higher numbers of viable bacteria with a greater level of repeatability than plate counting (RSD of 6.82 and 13.14% respectively). Across all multi-species blends tested, viable cell input was more accurately recovered by flow cytometry (101.8 ± 6.95%) than plate counts (81.37 ± 16.03%). However, when certain probiotic mixtures contained preparations with high numbers of non-viable cells in their total population, flow cytometry had the potential for overestimation of the viable population. Nevertheless, the comparative plate counts of these mixtures were low and variable, thus supporting the use of flow cytometry for the enumeration of viable bacteria in mixed populations.
ISSN:1471-2180
1471-2180
DOI:10.1186/s12866-023-02792-2