This study investigated the colonization site and duration of L. plantarum CCFM8610 in the host intestine using in vivo and ex vivo fluorescence imaging. The results indicated that orally administered CCFM8610 primarily remained in the cecum and colon and could transiently survive 6 days in the intestine after cessation of administration. The impact of CCFM8610 on gut microbiota was described by 16s rRNA sequencing comparison of mouse and human fecal samples. The results showed that the intake of CCFM8610 would change the gut microbes at the phylum level, including an increase in the abundance of Firmicutes and a decrease in Fusobacteria, and some changes were persistent. Furthermore, based on functional predictions and metabolic results, ingestion of CCFM8610 primarily affects lipid metabolism pathways in healthy hosts especially bile acid metabolism. Overall, this study investigated the colonization ability of CCFM8610 in healthy hosts after ingestion and its effects on the host, providing an effective reference for the study of daily intake regimens of probiotics.