Extracellular DNA in the Chicken Cecal Environment

The presence and biological role of extracellular DNA (eDNA) have been explored in diverse microbial environments. Nonetheless, it has not been studied in the chicken gut microbiome. This study aims to investigate eDNA in the chicken cecum, analyzing cecal samples from broiler chickens using three p...

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Main Authors: Deborah Olubanjo, Artur Zbikowski, Bishnu Adhikari, Tieshan Jiang, Pallavi Singh, Seong W. Kang, Young Min Kwon
Format: Article
Language:English
Published: MDPI AG 2025-03-01
Series:Poultry
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Online Access:https://www.mdpi.com/2674-1164/4/1/14
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author Deborah Olubanjo
Artur Zbikowski
Bishnu Adhikari
Tieshan Jiang
Pallavi Singh
Seong W. Kang
Young Min Kwon
author_facet Deborah Olubanjo
Artur Zbikowski
Bishnu Adhikari
Tieshan Jiang
Pallavi Singh
Seong W. Kang
Young Min Kwon
author_sort Deborah Olubanjo
collection DOAJ
description The presence and biological role of extracellular DNA (eDNA) have been explored in diverse microbial environments. Nonetheless, it has not been studied in the chicken gut microbiome. This study aims to investigate eDNA in the chicken cecum, analyzing cecal samples from broiler chickens using three preparation methods: Whole Cecal Suspension (WCS), Washed Cell Pellets (WCP), and Cell-Free Supernatant (CFS). The 16S rRNA gene-based microbiota analysis revealed distinct microbial communities in CFS compared to WCS and WCP (<i>p</i> = 0.001). Notably, specific taxa, including <i>Anaerofilum</i>, <i>Anaerotruncus</i>, <i>Oscillospira, Syntrophomonas,</i> and <i>Delftia</i>, were enriched in CFS. Confocal fluorescence microscopy, employing stains such as Propidium Iodide (PI), GelGreen, and SYTO 9, confirmed the presence of eDNA with filaments observed in WCS and CFS. Colocalization of PI and GelGreen™ validated the extracellular nature of eDNA, while DNase I treatment selectively degraded eDNA, further confirming its extracellular nature. Our findings in this study highlight the presence of eDNA in the chicken cecal microbiome, and the presence of eDNA associated with specific taxonomic groups suggest that it might play a specific role in the biological function of the cecal microbiome, which warrants further investigation in the future.
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spelling doaj-art-93ad3009b04844f8bb419fd5d371d92e2025-08-20T03:43:14ZengMDPI AGPoultry2674-11642025-03-01411410.3390/poultry4010014Extracellular DNA in the Chicken Cecal EnvironmentDeborah Olubanjo0Artur Zbikowski1Bishnu Adhikari2Tieshan Jiang3Pallavi Singh4Seong W. Kang5Young Min Kwon6Center of Excellence for Poultry Science, University of Arkansas System Division of Agriculture, Fayetteville, AR 72701, USADepartment of Pathology & Veterinary Diagnostics, Institute of Veterinary Medicine, Warsaw University of Life Sciences, 02-776 Warsaw, PolandCenter of Excellence for Poultry Science, University of Arkansas System Division of Agriculture, Fayetteville, AR 72701, USACenter of Excellence for Poultry Science, University of Arkansas System Division of Agriculture, Fayetteville, AR 72701, USADepartment of Biological Sciences, Northern Illinois University, DeKalb, IL 60115, USACenter of Excellence for Poultry Science, University of Arkansas System Division of Agriculture, Fayetteville, AR 72701, USACenter of Excellence for Poultry Science, University of Arkansas System Division of Agriculture, Fayetteville, AR 72701, USAThe presence and biological role of extracellular DNA (eDNA) have been explored in diverse microbial environments. Nonetheless, it has not been studied in the chicken gut microbiome. This study aims to investigate eDNA in the chicken cecum, analyzing cecal samples from broiler chickens using three preparation methods: Whole Cecal Suspension (WCS), Washed Cell Pellets (WCP), and Cell-Free Supernatant (CFS). The 16S rRNA gene-based microbiota analysis revealed distinct microbial communities in CFS compared to WCS and WCP (<i>p</i> = 0.001). Notably, specific taxa, including <i>Anaerofilum</i>, <i>Anaerotruncus</i>, <i>Oscillospira, Syntrophomonas,</i> and <i>Delftia</i>, were enriched in CFS. Confocal fluorescence microscopy, employing stains such as Propidium Iodide (PI), GelGreen, and SYTO 9, confirmed the presence of eDNA with filaments observed in WCS and CFS. Colocalization of PI and GelGreen™ validated the extracellular nature of eDNA, while DNase I treatment selectively degraded eDNA, further confirming its extracellular nature. Our findings in this study highlight the presence of eDNA in the chicken cecal microbiome, and the presence of eDNA associated with specific taxonomic groups suggest that it might play a specific role in the biological function of the cecal microbiome, which warrants further investigation in the future.https://www.mdpi.com/2674-1164/4/1/14eDNAgut microbiomechickenceca16S rRNA geneconfocal fluorescence microscopy
spellingShingle Deborah Olubanjo
Artur Zbikowski
Bishnu Adhikari
Tieshan Jiang
Pallavi Singh
Seong W. Kang
Young Min Kwon
Extracellular DNA in the Chicken Cecal Environment
Poultry
eDNA
gut microbiome
chicken
ceca
16S rRNA gene
confocal fluorescence microscopy
title Extracellular DNA in the Chicken Cecal Environment
title_full Extracellular DNA in the Chicken Cecal Environment
title_fullStr Extracellular DNA in the Chicken Cecal Environment
title_full_unstemmed Extracellular DNA in the Chicken Cecal Environment
title_short Extracellular DNA in the Chicken Cecal Environment
title_sort extracellular dna in the chicken cecal environment
topic eDNA
gut microbiome
chicken
ceca
16S rRNA gene
confocal fluorescence microscopy
url https://www.mdpi.com/2674-1164/4/1/14
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