Cargando…

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Tracey, Harry, Coates, Niall, Hulme, Eleri, John, Daniel, Michael, Daryn Robert, Plummer, Susan Frances
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9969615/
https://www.ncbi.nlm.nih.gov/pubmed/36849905
http://dx.doi.org/10.1186/s12866-023-02792-2
_version_ 1784897769207496704
author Tracey, Harry
Coates, Niall
Hulme, Eleri
John, Daniel
Michael, Daryn Robert
Plummer, Susan Frances
author_facet Tracey, Harry
Coates, Niall
Hulme, Eleri
John, Daniel
Michael, Daryn Robert
Plummer, Susan Frances
author_sort Tracey, Harry
collection PubMed
description 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-023-02792-2.
format Online
Article
Text
id pubmed-9969615
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-99696152023-02-28 Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry Tracey, Harry Coates, Niall Hulme, Eleri John, Daniel Michael, Daryn Robert Plummer, Susan Frances BMC Microbiol Research 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. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12866-023-02792-2. BioMed Central 2023-02-27 /pmc/articles/PMC9969615/ /pubmed/36849905 http://dx.doi.org/10.1186/s12866-023-02792-2 Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Tracey, Harry
Coates, Niall
Hulme, Eleri
John, Daniel
Michael, Daryn Robert
Plummer, Susan Frances
Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
title Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
title_full Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
title_fullStr Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
title_full_unstemmed Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
title_short Insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
title_sort insights into the enumeration of mixtures of probiotic bacteria by flow cytometry
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9969615/
https://www.ncbi.nlm.nih.gov/pubmed/36849905
http://dx.doi.org/10.1186/s12866-023-02792-2
work_keys_str_mv AT traceyharry insightsintotheenumerationofmixturesofprobioticbacteriabyflowcytometry
AT coatesniall insightsintotheenumerationofmixturesofprobioticbacteriabyflowcytometry
AT hulmeeleri insightsintotheenumerationofmixturesofprobioticbacteriabyflowcytometry
AT johndaniel insightsintotheenumerationofmixturesofprobioticbacteriabyflowcytometry
AT michaeldarynrobert insightsintotheenumerationofmixturesofprobioticbacteriabyflowcytometry
AT plummersusanfrances insightsintotheenumerationofmixturesofprobioticbacteriabyflowcytometry