Cargando…

Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri

Optimisation of cultivation conditions in the industrial production of probiotics is crucial to reach a high-quality product with retained probiotic functionality. Flow cytometry-based descriptors of bacterial morphology may be used as markers to estimate physiological fitness during cultivation, an...

Descripción completa

Detalles Bibliográficos
Autores principales: Rao, Nikhil Seshagiri, Lundberg, Ludwig, Palmkron, Shuai, Håkansson, Sebastian, Bergenståhl, Björn, Carlquist, Magnus
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8651721/
https://www.ncbi.nlm.nih.gov/pubmed/34876641
http://dx.doi.org/10.1038/s41598-021-02919-3
_version_ 1784611460046913536
author Rao, Nikhil Seshagiri
Lundberg, Ludwig
Palmkron, Shuai
Håkansson, Sebastian
Bergenståhl, Björn
Carlquist, Magnus
author_facet Rao, Nikhil Seshagiri
Lundberg, Ludwig
Palmkron, Shuai
Håkansson, Sebastian
Bergenståhl, Björn
Carlquist, Magnus
author_sort Rao, Nikhil Seshagiri
collection PubMed
description Optimisation of cultivation conditions in the industrial production of probiotics is crucial to reach a high-quality product with retained probiotic functionality. Flow cytometry-based descriptors of bacterial morphology may be used as markers to estimate physiological fitness during cultivation, and can be applied for online monitoring to avoid suboptimal growth. In the current study, the effects of temperature, initial pH and oxygen levels on cell growth and cell size distributions of Limosilactobacillus reuteri DSM 17938 were measured using multivariate flow cytometry. A pleomorphic behaviour was evident from the measurements of light scatter and pulse width distributions. A pattern of high growth yielding smaller cells and less heterogeneous populations could be observed. Analysis of pulse width distributions revealed significant morphological heterogeneities within the bacterial cell population under non-optimal growth conditions, and pointed towards low temperature, high initial pH, and high oxygen levels all being triggers for changes in morphology towards cell chain formation. However, cell size did not correlate to survivability after freeze-thaw or freeze-drying stress, indicating that it is not a key determinant for physical stress tolerance. The fact that L. reuteri morphology varies depending on cultivation conditions suggests that it can be used as marker for estimating physiological fitness and responses to its environment.
format Online
Article
Text
id pubmed-8651721
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-86517212021-12-08 Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri Rao, Nikhil Seshagiri Lundberg, Ludwig Palmkron, Shuai Håkansson, Sebastian Bergenståhl, Björn Carlquist, Magnus Sci Rep Article Optimisation of cultivation conditions in the industrial production of probiotics is crucial to reach a high-quality product with retained probiotic functionality. Flow cytometry-based descriptors of bacterial morphology may be used as markers to estimate physiological fitness during cultivation, and can be applied for online monitoring to avoid suboptimal growth. In the current study, the effects of temperature, initial pH and oxygen levels on cell growth and cell size distributions of Limosilactobacillus reuteri DSM 17938 were measured using multivariate flow cytometry. A pleomorphic behaviour was evident from the measurements of light scatter and pulse width distributions. A pattern of high growth yielding smaller cells and less heterogeneous populations could be observed. Analysis of pulse width distributions revealed significant morphological heterogeneities within the bacterial cell population under non-optimal growth conditions, and pointed towards low temperature, high initial pH, and high oxygen levels all being triggers for changes in morphology towards cell chain formation. However, cell size did not correlate to survivability after freeze-thaw or freeze-drying stress, indicating that it is not a key determinant for physical stress tolerance. The fact that L. reuteri morphology varies depending on cultivation conditions suggests that it can be used as marker for estimating physiological fitness and responses to its environment. Nature Publishing Group UK 2021-12-07 /pmc/articles/PMC8651721/ /pubmed/34876641 http://dx.doi.org/10.1038/s41598-021-02919-3 Text en © The Author(s) 2021 https://creativecommons.org/licenses/by/4.0/Open Access This 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/) .
spellingShingle Article
Rao, Nikhil Seshagiri
Lundberg, Ludwig
Palmkron, Shuai
Håkansson, Sebastian
Bergenståhl, Björn
Carlquist, Magnus
Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri
title Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri
title_full Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri
title_fullStr Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri
title_full_unstemmed Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri
title_short Flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of Limosilactobacillus reuteri
title_sort flow cytometric analysis reveals culture condition dependent variations in phenotypic heterogeneity of limosilactobacillus reuteri
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8651721/
https://www.ncbi.nlm.nih.gov/pubmed/34876641
http://dx.doi.org/10.1038/s41598-021-02919-3
work_keys_str_mv AT raonikhilseshagiri flowcytometricanalysisrevealscultureconditiondependentvariationsinphenotypicheterogeneityoflimosilactobacillusreuteri
AT lundbergludwig flowcytometricanalysisrevealscultureconditiondependentvariationsinphenotypicheterogeneityoflimosilactobacillusreuteri
AT palmkronshuai flowcytometricanalysisrevealscultureconditiondependentvariationsinphenotypicheterogeneityoflimosilactobacillusreuteri
AT hakanssonsebastian flowcytometricanalysisrevealscultureconditiondependentvariationsinphenotypicheterogeneityoflimosilactobacillusreuteri
AT bergenstahlbjorn flowcytometricanalysisrevealscultureconditiondependentvariationsinphenotypicheterogeneityoflimosilactobacillusreuteri
AT carlquistmagnus flowcytometricanalysisrevealscultureconditiondependentvariationsinphenotypicheterogeneityoflimosilactobacillusreuteri