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 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 (p = 0.001). Notably, specific taxa, including Anaerofilum, Anaerotruncus, Oscillospira, Syntrophomonas, and Delftia, 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.