Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times

The aim of this study was to investigate the effects of storage temperature and preservation time on the microbial diversity and community composition of rumen fluid. Rumen fluid samples were collected from six Hu sheep fed on a high-forage diet and stored at −80 °C and −20 °C for intervals of 0, 7,...

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Main Authors: Chang Liu, Jin Cheng, Yunong Xie, Kehui Ouyang, Mingren Qu, Ke Pan, Qinghua Qiu
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Veterinary Sciences
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Online Access:https://www.mdpi.com/2306-7381/12/3/234
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author Chang Liu
Jin Cheng
Yunong Xie
Kehui Ouyang
Mingren Qu
Ke Pan
Qinghua Qiu
author_facet Chang Liu
Jin Cheng
Yunong Xie
Kehui Ouyang
Mingren Qu
Ke Pan
Qinghua Qiu
author_sort Chang Liu
collection DOAJ
description The aim of this study was to investigate the effects of storage temperature and preservation time on the microbial diversity and community composition of rumen fluid. Rumen fluid samples were collected from six Hu sheep fed on a high-forage diet and stored at −80 °C and −20 °C for intervals of 0, 7, 14, 30, 60, 120, and 240 days. DNA was extracted at each time point for 16S rRNA gene sequencing to evaluate the rumen microbial diversity and community composition. The results showed that storage temperature affected only the relative abundance of Proteobacteria, with no substantial impact on alpha-diversity or other microbial groups (<i>p</i> > 0.05), and no significant interaction effects were observed between storage temperature and preservation time (<i>p</i> > 0.05). Alpha-diversity indices such as Chao1, observed species, and PD whole tree showed dynamic changes after 7 days of storage, while the relative abundances of Verrucomicrobiota and <i>Christensenellaceae R-7 group</i>, as well as the energy metabolism metabolic pathway, exhibited significant alterations after 14 days of storage (<i>p</i> < 0.05). Notably, Patescibacteria, <i>Rikenellaceae RC9 gut group</i>, and <i>Veillonellaceae UCG-001</i> abundances demonstrated significant changes after 240 days of storage (<i>p</i> < 0.05). Both principal coordinates analysis (PCoA) and non-metric multidimensional scaling (NMDS) showed distinct overlaps. This study suggests that storing rumen fluid at −80 °C and −20 °C does not influence rumen microbial diversity and community composition, whereas the storage time significantly impacts these factors, with most differences emerging after 14 days of preservation. Consequently, it is advised that the analysis of microbial diversity and community composition in rumen fluid samples be conducted within 14 days post-collection.
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publishDate 2025-03-01
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series Veterinary Sciences
spelling doaj-art-2f8beeb6a3be462597c1a78babe68c452025-08-20T01:49:53ZengMDPI AGVeterinary Sciences2306-73812025-03-0112323410.3390/vetsci12030234Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation TimesChang Liu0Jin Cheng1Yunong Xie2Kehui Ouyang3Mingren Qu4Ke Pan5Qinghua Qiu6Jiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaJiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaJiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaJiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaJiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaJiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaJiangxi Province Key Laboratory of Animal Nutrition and Feed, College of Animal Science and Technology, Jiangxi Agricultural University, Nanchang 330045, ChinaThe aim of this study was to investigate the effects of storage temperature and preservation time on the microbial diversity and community composition of rumen fluid. Rumen fluid samples were collected from six Hu sheep fed on a high-forage diet and stored at −80 °C and −20 °C for intervals of 0, 7, 14, 30, 60, 120, and 240 days. DNA was extracted at each time point for 16S rRNA gene sequencing to evaluate the rumen microbial diversity and community composition. The results showed that storage temperature affected only the relative abundance of Proteobacteria, with no substantial impact on alpha-diversity or other microbial groups (<i>p</i> > 0.05), and no significant interaction effects were observed between storage temperature and preservation time (<i>p</i> > 0.05). Alpha-diversity indices such as Chao1, observed species, and PD whole tree showed dynamic changes after 7 days of storage, while the relative abundances of Verrucomicrobiota and <i>Christensenellaceae R-7 group</i>, as well as the energy metabolism metabolic pathway, exhibited significant alterations after 14 days of storage (<i>p</i> < 0.05). Notably, Patescibacteria, <i>Rikenellaceae RC9 gut group</i>, and <i>Veillonellaceae UCG-001</i> abundances demonstrated significant changes after 240 days of storage (<i>p</i> < 0.05). Both principal coordinates analysis (PCoA) and non-metric multidimensional scaling (NMDS) showed distinct overlaps. This study suggests that storing rumen fluid at −80 °C and −20 °C does not influence rumen microbial diversity and community composition, whereas the storage time significantly impacts these factors, with most differences emerging after 14 days of preservation. Consequently, it is advised that the analysis of microbial diversity and community composition in rumen fluid samples be conducted within 14 days post-collection.https://www.mdpi.com/2306-7381/12/3/234bacterial community compositionbacterial diversitypreservation timequorum sensingrumen fluidstorage temperature
spellingShingle Chang Liu
Jin Cheng
Yunong Xie
Kehui Ouyang
Mingren Qu
Ke Pan
Qinghua Qiu
Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times
Veterinary Sciences
bacterial community composition
bacterial diversity
preservation time
quorum sensing
rumen fluid
storage temperature
title Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times
title_full Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times
title_fullStr Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times
title_full_unstemmed Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times
title_short Dynamic Changes in Rumen Microbial Diversity and Community Composition Within Rumen Fluid in Response to Various Storage Temperatures and Preservation Times
title_sort dynamic changes in rumen microbial diversity and community composition within rumen fluid in response to various storage temperatures and preservation times
topic bacterial community composition
bacterial diversity
preservation time
quorum sensing
rumen fluid
storage temperature
url https://www.mdpi.com/2306-7381/12/3/234
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