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Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies

The number of live bacterial cells is the most used parameter to assess the quality of finished probiotic products. Plate counting (PC) is the standard method in industry to enumerate cells. Application of PC implies critical aspects related to the selection of optimal nutrient media and growth cond...

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Autores principales: Visciglia, Annalisa, Allesina, Serena, Amoruso, Angela, De Prisco, Annachiara, Dhir, Raja, Bron, Peter A., Pane, Marco
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9672074/
https://www.ncbi.nlm.nih.gov/pubmed/36406457
http://dx.doi.org/10.3389/fmicb.2022.989563
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author Visciglia, Annalisa
Allesina, Serena
Amoruso, Angela
De Prisco, Annachiara
Dhir, Raja
Bron, Peter A.
Pane, Marco
author_facet Visciglia, Annalisa
Allesina, Serena
Amoruso, Angela
De Prisco, Annachiara
Dhir, Raja
Bron, Peter A.
Pane, Marco
author_sort Visciglia, Annalisa
collection PubMed
description The number of live bacterial cells is the most used parameter to assess the quality of finished probiotic products. Plate counting (PC) is the standard method in industry to enumerate cells. Application of PC implies critical aspects related to the selection of optimal nutrient media and growth conditions and underestimation of viable but not cultivable (VBNC) cells. Flow-cytometry (FC) is a culture-independent methodology having the potential to selectively enumerate live, damaged, and dead cells representing a powerful tool for in-depth monitoring of probiotic products. We monitored the shelf life of a clinical batch of a synbiotic composition PDS-08 targeting the pediatric population by means of PC and FC according to International Conference on Harmonization (ICH) pharma guidelines testing the Arrhenius model as predictive tool; PC enumeration revealed higher destruction rate than FC suggesting a faster reduction in cultivability than membrane integrity and thus a possible shift of the bacteria into a VBNC status. PDS-08 maintained acidification capability over time, when re-suspended in nutrient medium, even in samples tested sub-optimally for CFU detection (below 1 billion cells/dose). Due to similar kinetics described by the study of metabolic activity and membrane integrity, FC might be suggested as a valid tool for the study of functional stability of a probiotic product.
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spelling pubmed-96720742022-11-19 Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies Visciglia, Annalisa Allesina, Serena Amoruso, Angela De Prisco, Annachiara Dhir, Raja Bron, Peter A. Pane, Marco Front Microbiol Microbiology The number of live bacterial cells is the most used parameter to assess the quality of finished probiotic products. Plate counting (PC) is the standard method in industry to enumerate cells. Application of PC implies critical aspects related to the selection of optimal nutrient media and growth conditions and underestimation of viable but not cultivable (VBNC) cells. Flow-cytometry (FC) is a culture-independent methodology having the potential to selectively enumerate live, damaged, and dead cells representing a powerful tool for in-depth monitoring of probiotic products. We monitored the shelf life of a clinical batch of a synbiotic composition PDS-08 targeting the pediatric population by means of PC and FC according to International Conference on Harmonization (ICH) pharma guidelines testing the Arrhenius model as predictive tool; PC enumeration revealed higher destruction rate than FC suggesting a faster reduction in cultivability than membrane integrity and thus a possible shift of the bacteria into a VBNC status. PDS-08 maintained acidification capability over time, when re-suspended in nutrient medium, even in samples tested sub-optimally for CFU detection (below 1 billion cells/dose). Due to similar kinetics described by the study of metabolic activity and membrane integrity, FC might be suggested as a valid tool for the study of functional stability of a probiotic product. Frontiers Media S.A. 2022-11-04 /pmc/articles/PMC9672074/ /pubmed/36406457 http://dx.doi.org/10.3389/fmicb.2022.989563 Text en Copyright © 2022 Visciglia, Allesina, Amoruso, De Prisco, Dhir, Bron and Pane. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Microbiology
Visciglia, Annalisa
Allesina, Serena
Amoruso, Angela
De Prisco, Annachiara
Dhir, Raja
Bron, Peter A.
Pane, Marco
Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
title Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
title_full Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
title_fullStr Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
title_full_unstemmed Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
title_short Assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
title_sort assessment of shelf-life and metabolic viability of a multi-strain synbiotic using standard and innovative enumeration technologies
topic Microbiology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9672074/
https://www.ncbi.nlm.nih.gov/pubmed/36406457
http://dx.doi.org/10.3389/fmicb.2022.989563
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