Abstract:[Objective] Intestinal bacteria play an important role in host metabolism and health. It has been reported that the intestinal flora of Gansu zokor is related to regional, gender and seasonal factors, however, it is not clear whether there is a difference in the intestinal bacterial of Gansu zokor feeding with different feeds under wild and artificial feeding conditions. [Methods] In this study, we selected alfalfa, the roots of Mongolian, Chinese pine and spruce for feeding groups, wild Gansu zokors captured in the same habitat as controls. Bacterial diversity of digestive tract contents in Gansu zokor was analyzed by 16S rRNA V3-V4 high-throughput sequencing technology. [Results] Under the conditions of wild and artificial feeding, the dominant bacteria were both Firmicutes and Bacteroidetes, but the content was significantly different. Under artificial feeding conditions, the intestinal bacterial community structure of Mongolian pine, Chinese pine and spruce was similar, they all had significant difference with alfalfa and wild groups. The intestinal bacterial diversity of Mongolian pine, Chinese pine, spruce were higher than alfalfa and wild groups. [Conclusion] The results showed that there were significant differences in the composition and structure of intestinal bacterial of Gansu zokor under wild and artificial feeding conditions.