In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity

The gut microbiota (GM) is a large and diverse microbial community that plays essential roles in host health. The in vitro fermentation model of the fecal GM serves as a valuable complement to food and health research in both humans and animals. Despite advancements in standardized protocols for cul...

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Main Authors: Qianle Ren, Yuling Li, Mingmei Duan, Jinjun Li, Fangshu Shi, Yun Zhou, Wanjing Hu, Junfu Mao, Xiaoqiong Li
Format: Article
Language:English
Published: Frontiers Media S.A. 2025-01-01
Series:Frontiers in Microbiology
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Online Access:https://www.frontiersin.org/articles/10.3389/fmicb.2025.1515865/full
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author Qianle Ren
Qianle Ren
Yuling Li
Mingmei Duan
Jinjun Li
Fangshu Shi
Yun Zhou
Wanjing Hu
Junfu Mao
Xiaoqiong Li
author_facet Qianle Ren
Qianle Ren
Yuling Li
Mingmei Duan
Jinjun Li
Fangshu Shi
Yun Zhou
Wanjing Hu
Junfu Mao
Xiaoqiong Li
author_sort Qianle Ren
collection DOAJ
description The gut microbiota (GM) is a large and diverse microbial community that plays essential roles in host health. The in vitro fermentation model of the fecal GM serves as a valuable complement to food and health research in both humans and animals. Despite advancements in standardized protocols for culturing human GM, research concerning animals—particularly companion animals—remains limited. This study aims to identify the optimal in vitro fermentation method for cat gut microbiota by comprehensively analyzing fecal microbiota and fermentation characteristics. We evaluated seven culture media previously used to simulate the gut microenvironment in humans, dogs, and cats: anaerobic medium base (AMB), Minimum medium (MM), Pet medium (PM), VI medium (VI), VL medium (VL), Yeast culture medium (JM), and yeast casitone fatty acid agar medium (YCFA). Fresh fecal samples were fermented in these media for 48 h, followed by 16S rRNA sequencing to assess bacterial community composition and targeted metabolite monitoring during fermentation. The results revealed that the substrate composition in the medium differentially impacts bacterial community structure and fermentation characteristics. High levels of carbon and nitrogen sources can substantially increase gas production, particularly CO2, while also significantly enhancing the production of short-chain fatty acids (SCFAs). Additionally, substrates with a high carbon-to-nitrogen ratio promote the production of more SCFAs and biogenic amines, and enrich the Bacteroidaceae family, even when the total substrate amount is lower. Comprehensive analysis of gut microbiota and metabolites reveals that PM medium effectively simulates a nutrient-deficient microenvironment in the cat gut during in vitro fermentation. This simulation maintains bacterial community stability and results in lower metabolite levels. Therefore, using PM medium to culture cat gut microbiota for 48 h, without focusing on specific bacterial genera, represents the most suitable in vitro model. This finding contributes to understanding the optimal conditions for simulate cat gut microbiota and may provide a new approach for investigating the food pharmaceuticals on the cat gut microbiota and related health.
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publisher Frontiers Media S.A.
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spelling doaj-art-7fc08fc22c624b62a849dd7cfc7770522025-01-29T06:45:49ZengFrontiers Media S.A.Frontiers in Microbiology1664-302X2025-01-011610.3389/fmicb.2025.15158651515865In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activityQianle Ren0Qianle Ren1Yuling Li2Mingmei Duan3Jinjun Li4Fangshu Shi5Yun Zhou6Wanjing Hu7Junfu Mao8Xiaoqiong Li9State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products & Food Sciences Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, ChinaCollege of Veterinary Medicine, Jilin University, Changchun, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products & Food Sciences Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products & Food Sciences Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products & Food Sciences Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products & Food Sciences Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, ChinaGuangzhou MYBAO Biotechnology Co., Ltd, Guangzhou, ChinaGuangzhou MYBAO Biotechnology Co., Ltd, Guangzhou, ChinaNew Ruipeng Pet Group Inc., Beijing, ChinaState Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products & Food Sciences Institute, Zhejiang Academy of Agricultural Sciences, Hangzhou, ChinaThe gut microbiota (GM) is a large and diverse microbial community that plays essential roles in host health. The in vitro fermentation model of the fecal GM serves as a valuable complement to food and health research in both humans and animals. Despite advancements in standardized protocols for culturing human GM, research concerning animals—particularly companion animals—remains limited. This study aims to identify the optimal in vitro fermentation method for cat gut microbiota by comprehensively analyzing fecal microbiota and fermentation characteristics. We evaluated seven culture media previously used to simulate the gut microenvironment in humans, dogs, and cats: anaerobic medium base (AMB), Minimum medium (MM), Pet medium (PM), VI medium (VI), VL medium (VL), Yeast culture medium (JM), and yeast casitone fatty acid agar medium (YCFA). Fresh fecal samples were fermented in these media for 48 h, followed by 16S rRNA sequencing to assess bacterial community composition and targeted metabolite monitoring during fermentation. The results revealed that the substrate composition in the medium differentially impacts bacterial community structure and fermentation characteristics. High levels of carbon and nitrogen sources can substantially increase gas production, particularly CO2, while also significantly enhancing the production of short-chain fatty acids (SCFAs). Additionally, substrates with a high carbon-to-nitrogen ratio promote the production of more SCFAs and biogenic amines, and enrich the Bacteroidaceae family, even when the total substrate amount is lower. Comprehensive analysis of gut microbiota and metabolites reveals that PM medium effectively simulates a nutrient-deficient microenvironment in the cat gut during in vitro fermentation. This simulation maintains bacterial community stability and results in lower metabolite levels. Therefore, using PM medium to culture cat gut microbiota for 48 h, without focusing on specific bacterial genera, represents the most suitable in vitro model. This finding contributes to understanding the optimal conditions for simulate cat gut microbiota and may provide a new approach for investigating the food pharmaceuticals on the cat gut microbiota and related health.https://www.frontiersin.org/articles/10.3389/fmicb.2025.1515865/fullcatgut microbiotain vitro fermentationtargeted metabolitesSCFAs
spellingShingle Qianle Ren
Qianle Ren
Yuling Li
Mingmei Duan
Jinjun Li
Fangshu Shi
Yun Zhou
Wanjing Hu
Junfu Mao
Xiaoqiong Li
In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity
Frontiers in Microbiology
cat
gut microbiota
in vitro fermentation
targeted metabolites
SCFAs
title In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity
title_full In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity
title_fullStr In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity
title_full_unstemmed In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity
title_short In vitro modeling of feline gut fermentation: a comprehensive analysis of fecal microbiota and metabolic activity
title_sort in vitro modeling of feline gut fermentation a comprehensive analysis of fecal microbiota and metabolic activity
topic cat
gut microbiota
in vitro fermentation
targeted metabolites
SCFAs
url https://www.frontiersin.org/articles/10.3389/fmicb.2025.1515865/full
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