metagenomic seq of Homo spaiens: fecal
To achieve species-level resolution, shotgun metagenomic sequencing was also performed on fecal DNA from the same participants. Briefly, DNA was fragmented to an average size of ~400 bp for paired-end library construction using the NEXTFLE Rapid DNA-Seq kit 38. Sequencing was carried out on the Illumina NovaSeq 6000 platform (Illumina Inc., San Diego, CA, USA). Metagenomic functional annotation was performed using HUMAnN3 with the ChocoPhlAn database for human samples, and DIAMOND (v2.0.13) against the NCBI NR protein database (version 20230830) for mouse samples. Then, functional analysis of differential microorganisms in the two groups was performed. All 16S rRNA and metagenomic data were processed using the Majorbio Cloud Platform (www.majorbio.com).