A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker

Frequent eructation in ruminant animals results in an exhaled blend of ruminal eructed and breath volatile organic compounds (VOC). The physiological distinction between the gas sources can limit the applicability of breath metabolomics (or breathomics) in describing the metabolic phenotype of cows....

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Main Authors: M.A. Barrientos-Blanco, U. Arshad, S. Giannoukos, M.Z. Islam, C. Kunz, R. Peng, S.E. Räisänen, R. Zenobi, M. Niu
Format: Article
Language:English
Published: Elsevier 2025-05-01
Series:JDS Communications
Online Access:http://www.sciencedirect.com/science/article/pii/S2666910225000432
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author M.A. Barrientos-Blanco
U. Arshad
S. Giannoukos
M.Z. Islam
C. Kunz
R. Peng
S.E. Räisänen
R. Zenobi
M. Niu
author_facet M.A. Barrientos-Blanco
U. Arshad
S. Giannoukos
M.Z. Islam
C. Kunz
R. Peng
S.E. Räisänen
R. Zenobi
M. Niu
author_sort M.A. Barrientos-Blanco
collection DOAJ
description Frequent eructation in ruminant animals results in an exhaled blend of ruminal eructed and breath volatile organic compounds (VOC). The physiological distinction between the gas sources can limit the applicability of breath metabolomics (or breathomics) in describing the metabolic phenotype of cows. The objective of this study was to establish a benchmark sampling method for collecting breath samples in dairy cows while they were not eructating. Twelve multiparous mid-lactation Holstein cows were enrolled to collect (1) breath (BR; bloodborne VOC exchanged at the lungs) and (2) ruminal exhaled (RE; a mixture of VOC from ruminal eructation and breaths during eructations) samples. Gas samples were collected using a head chamber (GreenFeed system) with real-time CH4 readings. By monitoring eructation events, a threshold of <150 mV CH4 was set to sample breath, and >250 mV was used to collect BR and RE. Both samples were analyzed using secondary electrospray ionization-high resolution MS (SESI-MS) and GC. Implementing CH4 as a marker resulted in 80% lower CH4 concentrations in BR compared with RE. Analysis using SESI-MS revealed a total of 324 and 242 features consistently identified across all periods of the study in [M-H]− and [M+H]+ MS ion mode, respectively, for BR and RE. In BR, 18 features exhibited greater concentrations, whereas 8 had a tendency to have greater concentrations compared with RE. In contrast, RE revealed 51 features with greater concentrations, and 13 with a tendency for greater concentrations compared with BR. Ruminal VFA acetate, propionate, and butyrate were 20.9%, 27.4%, and 32.7% greater in RE compared with BR, respectively. Lower CH4 levels in BR and the greater VFA concentrations in the RE validated the ability of the method to differentiate breath from ruminal eructed VOC. Our study established a method to distinguish and separately collect BR and RE samples in dairy cows. This advance shows the potential to use breathomics as a reliable and noninvasive tool for metabolic assessments in ruminant research.
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spelling doaj-art-9d79ac821dcd4115956ef85ceb649fdc2025-08-20T02:13:54ZengElsevierJDS Communications2666-91022025-05-016343844310.3168/jdsc.2024-0732A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a markerM.A. Barrientos-Blanco0U. Arshad1S. Giannoukos2M.Z. Islam3C. Kunz4R. Peng5S.E. Räisänen6R. Zenobi7M. Niu8Animal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandAnimal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandAnalytical Chemistry, Department of Chemistry and Applied Biosciences, ETH Zürich, 8093 Zürich, SwitzerlandAnimal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandAnimal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandAnimal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandAnimal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandAnalytical Chemistry, Department of Chemistry and Applied Biosciences, ETH Zürich, 8093 Zürich, SwitzerlandCorresponding author; Animal Nutrition, Institute of Agricultural Sciences, Department of Environmental Systems Science, ETH Zürich, 8092 Zürich, SwitzerlandFrequent eructation in ruminant animals results in an exhaled blend of ruminal eructed and breath volatile organic compounds (VOC). The physiological distinction between the gas sources can limit the applicability of breath metabolomics (or breathomics) in describing the metabolic phenotype of cows. The objective of this study was to establish a benchmark sampling method for collecting breath samples in dairy cows while they were not eructating. Twelve multiparous mid-lactation Holstein cows were enrolled to collect (1) breath (BR; bloodborne VOC exchanged at the lungs) and (2) ruminal exhaled (RE; a mixture of VOC from ruminal eructation and breaths during eructations) samples. Gas samples were collected using a head chamber (GreenFeed system) with real-time CH4 readings. By monitoring eructation events, a threshold of <150 mV CH4 was set to sample breath, and >250 mV was used to collect BR and RE. Both samples were analyzed using secondary electrospray ionization-high resolution MS (SESI-MS) and GC. Implementing CH4 as a marker resulted in 80% lower CH4 concentrations in BR compared with RE. Analysis using SESI-MS revealed a total of 324 and 242 features consistently identified across all periods of the study in [M-H]− and [M+H]+ MS ion mode, respectively, for BR and RE. In BR, 18 features exhibited greater concentrations, whereas 8 had a tendency to have greater concentrations compared with RE. In contrast, RE revealed 51 features with greater concentrations, and 13 with a tendency for greater concentrations compared with BR. Ruminal VFA acetate, propionate, and butyrate were 20.9%, 27.4%, and 32.7% greater in RE compared with BR, respectively. Lower CH4 levels in BR and the greater VFA concentrations in the RE validated the ability of the method to differentiate breath from ruminal eructed VOC. Our study established a method to distinguish and separately collect BR and RE samples in dairy cows. This advance shows the potential to use breathomics as a reliable and noninvasive tool for metabolic assessments in ruminant research.http://www.sciencedirect.com/science/article/pii/S2666910225000432
spellingShingle M.A. Barrientos-Blanco
U. Arshad
S. Giannoukos
M.Z. Islam
C. Kunz
R. Peng
S.E. Räisänen
R. Zenobi
M. Niu
A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
JDS Communications
title A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
title_full A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
title_fullStr A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
title_full_unstemmed A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
title_short A sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
title_sort sampling method for differentiating breath and ruminal exhaled volatile organic compounds in dairy cows using methane as a marker
url http://www.sciencedirect.com/science/article/pii/S2666910225000432
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