94% of researchers rate our articles as excellent or good
Learn more about the work of our research integrity team to safeguard the quality of each article we publish.
Find out more
ORIGINAL RESEARCH article
Front. Microbiomes
Sec. Host and Microbe Associations
Volume 4 - 2025 | doi: 10.3389/frmbi.2025.1334775
This article is part of the Research TopicGut Microbiota and Its Importance on Human Health - the Need for Reliable Measurements to Assess the Microbial Gut Function and Its Correlated PathologiesView all 7 articles
Stability of Oral and Fecal Microbiome at Room Temperature: Impact on Diversity
Provisionally accepted
Blanca Rius-Sansalvador1,2,3
David Bars-Cortina1,2
Olfat Khannous4,5
Ainhoa Garcia-Serrano1,2Elisabet Guinó1,2,6
Ester Saus4,5
Toni Gabaldón4,5,7,8
Victor Moreno1,2,3,6*
Mireia Obon-Santacana1,2,6*- 1Institut d'Investigacio Biomedica de Bellvitge (IDIBELL), Barcelona, Catalonia, Spain
- 2Catalan Institute of Oncology, Barcelona, Catalonia, Spain
- 3University of Barcelona, Barcelona, Catalonia, Spain
- 4Barcelona Supercomputing Center, Barcelona, Catalonia, Spain
- 5Institute for Research in Biomedicine, Barcelona, Catalonia, Spain
- 6Center for Biomedical Research in Epidemiology and Public Health Network, Carlos III Health Institute (ISCIII), Madrid, Madrid, Spain
- 7Catalan Institution for Research and Advanced Studies (ICREA), Barcelona, Catalonia, Spain
- 8Networked Biomedical Research Center for Infectious Diseases (CIBERINFEC), Madrid, Asturias, Spain
Select one of your emails
You have multiple emails registered with Frontiers:
Notify me on publication
Please enter your email address:
If you already have an account, please login
You don't have a Frontiers account ? You can register here
When collecting oral and fecal samples for large epidemiological microbiome studies, optimal storage conditions such as immediate freezing, are not always feasible. It is fundamental to study the impact of temporary room temperature (RT) storage and shipping on the microbiome diversity obtained in different types of samples. We performed a pilot study aimed at validating the sampling protocol based on the viability of the 16S rRNA gene sequencing in microbiome samples.Fecal and oral samples from five participants were collected and preserved in different conditions: a) 70% ethanol; b) in a FIT tube for stool samples; and c) in a chlorhexidine solution for oral wash samples. Four aliquots were prepared per sample, which were stored at RT, and frozen at days 0, 5, 10 and 15, respectively. In terms of alpha diversity, the maximum average decrease in 5 days was 0.3%, 1.6% and 1.7% for oral, stool in ethanol and stool in FIT, respectively. Furthermore, the relative abundances of the most important phyla and orders remained stable over the two weeks.Our findings suggest that microbiome diversity is remarkably resilient, with fecal and oral samples stored at RT in 70% ethanol, chlorhexidine, and FIT tubes showing no substantial changes over 15 days. These results support the feasibility of large-scale microbiome studies relying on delayed sample processing.
Keywords: oral microbiome, fecal microbiome, Stability study, GCAT, 16S, Pilot Study
Received: 07 Nov 2023; Accepted: 09 Apr 2025.
Copyright: © 2025 Rius-Sansalvador, Bars-Cortina, Khannous, Garcia-Serrano, Guinó, Saus, Gabaldón, Moreno and Obon-Santacana. 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) or licensor 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.
* Correspondence:
Victor Moreno, Catalan Institute of Oncology, Barcelona, 08908, Catalonia, Spain
Mireia Obon-Santacana, Catalan Institute of Oncology, Barcelona, 08908, Catalonia, Spain
Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.