Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments

The gut microbiota has a profound effect on human health and is modulated by food and bioactive compounds. To study such interaction, in vitro batch fermentations are performed with fecal material, and some experimental designs may require that such fermentations be performed with previously frozen...

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Autores: Perez-Burillo, S, Hinojosa-Nogueira, D, Navajas-Porras, B, Blasco, T, Balzerani, F, Lerma-Aguilera, A, Leon, D, Pastoriza, S, Apaolaza, I, Planes, FJ, Francino, MP, Rufian-Henares, JA
Tipo de documento: artigo
Estado:Versão publicada
Data de publicação:2021
País:España
Recursos:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
Repositório:r-FISABIO. Repositorio Institucional de Producción Científica
OAI Identifier:oai:fisabio.fundanetsuite.com:p12924
Acesso em linha:https://fisabio.portalinvestigacion.com/publicaciones/12924
Access Level:Acceso aberto
Palavra-chave:gut microbiota
freezing
storage
foods
bioactive compounds
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spelling Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation ExperimentsPerez-Burillo, SHinojosa-Nogueira, DNavajas-Porras, BBlasco, TBalzerani, FLerma-Aguilera, ALeon, DPastoriza, SApaolaza, IPlanes, FJFrancino, MPRufian-Henares, JAgut microbiotafreezingstoragefoodsbioactive compoundsThe gut microbiota has a profound effect on human health and is modulated by food and bioactive compounds. To study such interaction, in vitro batch fermentations are performed with fecal material, and some experimental designs may require that such fermentations be performed with previously frozen stools. Although it is known that freezing fecal material does not alter the composition of the microbial community in 16S rRNA gene amplicon and metagenomic sequencing studies, it is not known whether the microbial community in frozen samples could still be used for in vitro fermentations. To explore this, we undertook a pilot study in which in vitro fermentations were performed with fecal material from celiac, cow's milk allergic, obese, or lean children that was frozen (or not) with 20% glycerol. Before fermentation, the fecal material was incubated in a nutritious medium for 6 days, with the aim of giving the microbial community time to recover from the effects of freezing. An aliquot was taken daily from the stabilization vessel and used for the in vitro batch fermentation of lentils. The microbial community structure was significantly different between fresh and frozen samples, but the variation introduced by freezing a sample was always smaller than the variation among individuals, both before and after fermentation. Moreover, the potential functionality (as determined in silico by a genome-scaled metabolic reconstruction) did not differ significantly, possibly due to functional redundancy. The most affected genus was Bacteroides, a fiber degrader. In conclusion, if frozen fecal material is to be used for in vitro fermentation purposes, our preliminary analyses indicate that the functionality of microbial communities can be preserved after stabilization.MDPI2021info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttps://fisabio.portalinvestigacion.com/publicaciones/12924NutrientsISSN: 20726643reponame:r-FISABIO. Repositorio Institucional de Producción Científicainstname:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)Inglésinfo:eu-repo/semantics/openAccessoai:fisabio.fundanetsuite.com:p129242026-06-11T12:45:17Z
dc.title.none.fl_str_mv Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
title Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
spellingShingle Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
Perez-Burillo, S
gut microbiota
freezing
storage
foods
bioactive compounds
title_short Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
title_full Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
title_fullStr Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
title_full_unstemmed Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
title_sort Effect of Freezing on Gut Microbiota Composition and Functionality for In Vitro Fermentation Experiments
dc.creator.none.fl_str_mv Perez-Burillo, S
Hinojosa-Nogueira, D
Navajas-Porras, B
Blasco, T
Balzerani, F
Lerma-Aguilera, A
Leon, D
Pastoriza, S
Apaolaza, I
Planes, FJ
Francino, MP
Rufian-Henares, JA
author Perez-Burillo, S
author_facet Perez-Burillo, S
Hinojosa-Nogueira, D
Navajas-Porras, B
Blasco, T
Balzerani, F
Lerma-Aguilera, A
Leon, D
Pastoriza, S
Apaolaza, I
Planes, FJ
Francino, MP
Rufian-Henares, JA
author_role author
author2 Hinojosa-Nogueira, D
Navajas-Porras, B
Blasco, T
Balzerani, F
Lerma-Aguilera, A
Leon, D
Pastoriza, S
Apaolaza, I
Planes, FJ
Francino, MP
Rufian-Henares, JA
author2_role author
author
author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv gut microbiota
freezing
storage
foods
bioactive compounds
topic gut microbiota
freezing
storage
foods
bioactive compounds
description The gut microbiota has a profound effect on human health and is modulated by food and bioactive compounds. To study such interaction, in vitro batch fermentations are performed with fecal material, and some experimental designs may require that such fermentations be performed with previously frozen stools. Although it is known that freezing fecal material does not alter the composition of the microbial community in 16S rRNA gene amplicon and metagenomic sequencing studies, it is not known whether the microbial community in frozen samples could still be used for in vitro fermentations. To explore this, we undertook a pilot study in which in vitro fermentations were performed with fecal material from celiac, cow's milk allergic, obese, or lean children that was frozen (or not) with 20% glycerol. Before fermentation, the fecal material was incubated in a nutritious medium for 6 days, with the aim of giving the microbial community time to recover from the effects of freezing. An aliquot was taken daily from the stabilization vessel and used for the in vitro batch fermentation of lentils. The microbial community structure was significantly different between fresh and frozen samples, but the variation introduced by freezing a sample was always smaller than the variation among individuals, both before and after fermentation. Moreover, the potential functionality (as determined in silico by a genome-scaled metabolic reconstruction) did not differ significantly, possibly due to functional redundancy. The most affected genus was Bacteroides, a fiber degrader. In conclusion, if frozen fecal material is to be used for in vitro fermentation purposes, our preliminary analyses indicate that the functionality of microbial communities can be preserved after stabilization.
publishDate 2021
dc.date.none.fl_str_mv 2021
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv https://fisabio.portalinvestigacion.com/publicaciones/12924
url https://fisabio.portalinvestigacion.com/publicaciones/12924
dc.language.none.fl_str_mv Inglés
language_invalid_str_mv Inglés
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
eu_rights_str_mv openAccess
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv Nutrients
ISSN: 20726643
reponame:r-FISABIO. Repositorio Institucional de Producción Científica
instname:Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
instname_str Fundación para el Fomento de la Investigación Sanitaria y Biomédica de la Comunitat Valenciana (FISABIO)
reponame_str r-FISABIO. Repositorio Institucional de Producción Científica
collection r-FISABIO. Repositorio Institucional de Producción Científica
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repository.mail.fl_str_mv
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