Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies

The gut microbiome is vital for health and affects gastrointestinal, systemic, and neurological functions. In dogs, fecal samples provide an effective mechanism to assess the gut microbiota as they are non-invasive, easily obtained, and representative of the gut microbiome. However, traditional meth...

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Main Authors: David Atuahene, Luke Wolfe, Oona Elisabet Vanhatalo, Benjamin T. Veenstra, Joseph H. Skarlupka, Katie L. Anderson, Garret Suen, Giorgia Meineri, Jessica C. Pritchard
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-07-01
Series:Frontiers in Veterinary Science
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Online Access:https://www.frontiersin.org/articles/10.3389/fvets.2025.1632686/full
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author David Atuahene
David Atuahene
Luke Wolfe
Oona Elisabet Vanhatalo
Benjamin T. Veenstra
Joseph H. Skarlupka
Katie L. Anderson
Garret Suen
Giorgia Meineri
Jessica C. Pritchard
author_facet David Atuahene
David Atuahene
Luke Wolfe
Oona Elisabet Vanhatalo
Benjamin T. Veenstra
Joseph H. Skarlupka
Katie L. Anderson
Garret Suen
Giorgia Meineri
Jessica C. Pritchard
author_sort David Atuahene
collection DOAJ
description The gut microbiome is vital for health and affects gastrointestinal, systemic, and neurological functions. In dogs, fecal samples provide an effective mechanism to assess the gut microbiota as they are non-invasive, easily obtained, and representative of the gut microbiome. However, traditional methods usually require sampling across three consecutive days per time point to minimize the presumed variation in the gut microbiome. Here, we sought to investigate whether the gut microbiome obtained from a single-day fecal sampling is reflective of the microbiome obtained from three-day collections. To accomplish this, we collected fecal samples from 12 dogs over 3 days and compared each single-day microbiome against the combined microbiotas of the three-day samples. We found no significant daily variation in the gut microbiota, as determined by two one-sided tests of equivalence (TOST) analysis, indicating that there are little to no day-to-day changes in the microbiota. Further microbial comparisons using PERMANOVA (p = 0.98) and non-metric multidimensional scaling also showed no significant differences in the microbial composition across the sampled days. Taken together, our findings suggest that a single sample can represent the gut microbiome as accurately as samples obtained across three consecutive days. As such, a single-day sampling approach can be used in dog microbiome studies, which would reduce both labor and costs while preserving overall data quality.
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spelling doaj-art-81ee0e8593ee4f42b2fa4922fe9b10f32025-08-20T03:09:39ZengFrontiers Media S.A.Frontiers in Veterinary Science2297-17692025-07-011210.3389/fvets.2025.16326861632686Limited day-to-day variation in the canine gut microbiota: implications for microbiome studiesDavid Atuahene0David Atuahene1Luke Wolfe2Oona Elisabet Vanhatalo3Benjamin T. Veenstra4Joseph H. Skarlupka5Katie L. Anderson6Garret Suen7Giorgia Meineri8Jessica C. Pritchard9Department of Veterinary Sciences, School of Agriculture and Veterinary Medicine, University of Turin, Turin, ItalyDepartment of Bacteriology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Bacteriology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Medical Sciences, University of Wisconsin-Madison School of Veterinary Medicine, Madison, WI, United StatesDepartment of Medical Sciences, University of Wisconsin-Madison School of Veterinary Medicine, Madison, WI, United StatesDepartment of Bacteriology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Medical Sciences, University of Wisconsin-Madison School of Veterinary Medicine, Madison, WI, United StatesDepartment of Bacteriology, University of Wisconsin-Madison, Madison, WI, United StatesDepartment of Veterinary Sciences, School of Agriculture and Veterinary Medicine, University of Turin, Turin, ItalyDepartment of Medical Sciences, University of Wisconsin-Madison School of Veterinary Medicine, Madison, WI, United StatesThe gut microbiome is vital for health and affects gastrointestinal, systemic, and neurological functions. In dogs, fecal samples provide an effective mechanism to assess the gut microbiota as they are non-invasive, easily obtained, and representative of the gut microbiome. However, traditional methods usually require sampling across three consecutive days per time point to minimize the presumed variation in the gut microbiome. Here, we sought to investigate whether the gut microbiome obtained from a single-day fecal sampling is reflective of the microbiome obtained from three-day collections. To accomplish this, we collected fecal samples from 12 dogs over 3 days and compared each single-day microbiome against the combined microbiotas of the three-day samples. We found no significant daily variation in the gut microbiota, as determined by two one-sided tests of equivalence (TOST) analysis, indicating that there are little to no day-to-day changes in the microbiota. Further microbial comparisons using PERMANOVA (p = 0.98) and non-metric multidimensional scaling also showed no significant differences in the microbial composition across the sampled days. Taken together, our findings suggest that a single sample can represent the gut microbiome as accurately as samples obtained across three consecutive days. As such, a single-day sampling approach can be used in dog microbiome studies, which would reduce both labor and costs while preserving overall data quality.https://www.frontiersin.org/articles/10.3389/fvets.2025.1632686/fullday-to-day variationdogs gut microbiomefecal samplingmicrobiome assessmentnon-invasive sampling
spellingShingle David Atuahene
David Atuahene
Luke Wolfe
Oona Elisabet Vanhatalo
Benjamin T. Veenstra
Joseph H. Skarlupka
Katie L. Anderson
Garret Suen
Giorgia Meineri
Jessica C. Pritchard
Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies
Frontiers in Veterinary Science
day-to-day variation
dogs gut microbiome
fecal sampling
microbiome assessment
non-invasive sampling
title Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies
title_full Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies
title_fullStr Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies
title_full_unstemmed Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies
title_short Limited day-to-day variation in the canine gut microbiota: implications for microbiome studies
title_sort limited day to day variation in the canine gut microbiota implications for microbiome studies
topic day-to-day variation
dogs gut microbiome
fecal sampling
microbiome assessment
non-invasive sampling
url https://www.frontiersin.org/articles/10.3389/fvets.2025.1632686/full
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