Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions

The objective of this study was to assess the effects of gallic acid (GA) on nutrient degradability, gas production, rumen fermentation, and the microbial community and its functions using in vitro fermentation methods. An in vitro experiment was conducted to test GA dose levels (0, 5, 10, 20, and 4...

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Main Authors: Wei Zhu, Jianjun Guo, Xin Li, Yan Li, Lianjie Song, Yunfei Li, Baoshan Feng, Xingnan Bao, Jianguo Li, Yanxia Gao, Hongjian Xu
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Language:English
Published: MDPI AG 2025-07-01
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Online Access:https://www.mdpi.com/2076-2615/15/13/1959
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author Wei Zhu
Jianjun Guo
Xin Li
Yan Li
Lianjie Song
Yunfei Li
Baoshan Feng
Xingnan Bao
Jianguo Li
Yanxia Gao
Hongjian Xu
author_facet Wei Zhu
Jianjun Guo
Xin Li
Yan Li
Lianjie Song
Yunfei Li
Baoshan Feng
Xingnan Bao
Jianguo Li
Yanxia Gao
Hongjian Xu
author_sort Wei Zhu
collection DOAJ
description The objective of this study was to assess the effects of gallic acid (GA) on nutrient degradability, gas production, rumen fermentation, and the microbial community and its functions using in vitro fermentation methods. An in vitro experiment was conducted to test GA dose levels (0, 5, 10, 20, and 40 mg/g DM) in the cow’s diet. Based on the results of nutrient degradability, gas production, and rumen fermentation, the control group (0 mg/g DM, CON) and the GA group (10 mg/g DM, GA) were selected for metagenomic analysis to further explore the microbial community and its functions. The degradability of dry matter and crude protein, as well as total gas production, CH<sub>4</sub> production, CH<sub>4</sub>/total gas, CO<sub>2</sub> production, and CO<sub>2</sub>/total gas, decreased quadratically (<i>p</i> < 0.05) with increasing GA doses, reaching their lowest levels at the 10 mg/g DM dose. Total volatile fatty acid (VFA) (<i>p</i> = 0.004), acetate (<i>p</i> = 0.03), and valerate (<i>p</i> = 0.03) exhibited quadratic decreases, while butyrate (<i>p</i> = 0.0006) showed a quadratic increase with increasing GA doses. The 10 mg/g DM dose group had the lowest levels of total VFA, acetate, and valerate, and the highest butyrate level compared to the other groups. The propionate (<i>p</i> = 0.03) and acetate-to-propionate ratio (<i>p</i> = 0.03) linearly decreased with increasing gallic acid inclusion. At the bacterial species level, GA supplementation significantly affected (<i>p</i> < 0.05) a total of 38 bacterial species. Among these, 29 species, such as <i>Prevotellasp.E15-22</i>, <i>bacteriumP3</i>, and <i>Alistipessp.CAG:435</i>, were less abundant in the GA group, while 9 species, including <i>Aristaeella_lactis</i> and <i>Aristaeella_hokkaidonensis</i>, were significantly more abundant in the GA group. At the archaeal species level, the relative abundances of <i>Methanobrevibacter_thaueri</i>, <i>Methanobrevibacter_boviskoreani</i>, and <i>Methanobrevibacter</i>sp.<i>AbM4</i> were significantly reduced (<i>p</i> < 0.05) by GA supplementation. Amino sugar and nucleotide sugar metabolism, Starch and sucrose metabolism, Glycolysis/Gluconeogenesis, and Pyruvate metabolismwere significantly enriched in the GA group (<i>p</i> < 0.05). Additionally, Alanine, aspartate and glutamate metabolism was also significantly enriched in the GA group (<i>p</i> < 0.05). GA use could potentially be an effective strategy for methane mitigation; however, further research is needed to assess its in vivo effects in dairy cows over a longer period.
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spelling doaj-art-ae87c42bd4654976861b171ef3c2a2d42025-08-20T03:28:34ZengMDPI AGAnimals2076-26152025-07-011513195910.3390/ani15131959Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic FunctionsWei Zhu0Jianjun Guo1Xin Li2Yan Li3Lianjie Song4Yunfei Li5Baoshan Feng6Xingnan Bao7Jianguo Li8Yanxia Gao9Hongjian Xu10College of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaChengde Academy of Agriculture and Forestry Sciences, Chengde 067000, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaCollege of Veterinary Medicine, Hebei Agricultural University, Baoding 071001, ChinaChengde Academy of Agriculture and Forestry Sciences, Chengde 067000, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaCollege of Animal Science and Technology, Hebei Agricultural University, Baoding 071001, ChinaThe objective of this study was to assess the effects of gallic acid (GA) on nutrient degradability, gas production, rumen fermentation, and the microbial community and its functions using in vitro fermentation methods. An in vitro experiment was conducted to test GA dose levels (0, 5, 10, 20, and 40 mg/g DM) in the cow’s diet. Based on the results of nutrient degradability, gas production, and rumen fermentation, the control group (0 mg/g DM, CON) and the GA group (10 mg/g DM, GA) were selected for metagenomic analysis to further explore the microbial community and its functions. The degradability of dry matter and crude protein, as well as total gas production, CH<sub>4</sub> production, CH<sub>4</sub>/total gas, CO<sub>2</sub> production, and CO<sub>2</sub>/total gas, decreased quadratically (<i>p</i> < 0.05) with increasing GA doses, reaching their lowest levels at the 10 mg/g DM dose. Total volatile fatty acid (VFA) (<i>p</i> = 0.004), acetate (<i>p</i> = 0.03), and valerate (<i>p</i> = 0.03) exhibited quadratic decreases, while butyrate (<i>p</i> = 0.0006) showed a quadratic increase with increasing GA doses. The 10 mg/g DM dose group had the lowest levels of total VFA, acetate, and valerate, and the highest butyrate level compared to the other groups. The propionate (<i>p</i> = 0.03) and acetate-to-propionate ratio (<i>p</i> = 0.03) linearly decreased with increasing gallic acid inclusion. At the bacterial species level, GA supplementation significantly affected (<i>p</i> < 0.05) a total of 38 bacterial species. Among these, 29 species, such as <i>Prevotellasp.E15-22</i>, <i>bacteriumP3</i>, and <i>Alistipessp.CAG:435</i>, were less abundant in the GA group, while 9 species, including <i>Aristaeella_lactis</i> and <i>Aristaeella_hokkaidonensis</i>, were significantly more abundant in the GA group. At the archaeal species level, the relative abundances of <i>Methanobrevibacter_thaueri</i>, <i>Methanobrevibacter_boviskoreani</i>, and <i>Methanobrevibacter</i>sp.<i>AbM4</i> were significantly reduced (<i>p</i> < 0.05) by GA supplementation. Amino sugar and nucleotide sugar metabolism, Starch and sucrose metabolism, Glycolysis/Gluconeogenesis, and Pyruvate metabolismwere significantly enriched in the GA group (<i>p</i> < 0.05). Additionally, Alanine, aspartate and glutamate metabolism was also significantly enriched in the GA group (<i>p</i> < 0.05). GA use could potentially be an effective strategy for methane mitigation; however, further research is needed to assess its in vivo effects in dairy cows over a longer period.https://www.mdpi.com/2076-2615/15/13/1959gallic acidmethane emissionrumen fermentationmetagenomics
spellingShingle Wei Zhu
Jianjun Guo
Xin Li
Yan Li
Lianjie Song
Yunfei Li
Baoshan Feng
Xingnan Bao
Jianguo Li
Yanxia Gao
Hongjian Xu
Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions
Animals
gallic acid
methane emission
rumen fermentation
metagenomics
title Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions
title_full Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions
title_fullStr Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions
title_full_unstemmed Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions
title_short Effects of Gallic Acid on In Vitro Ruminal Fermentation, Methane Emission, Microbial Composition, and Metabolic Functions
title_sort effects of gallic acid on in vitro ruminal fermentation methane emission microbial composition and metabolic functions
topic gallic acid
methane emission
rumen fermentation
metagenomics
url https://www.mdpi.com/2076-2615/15/13/1959
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