Fecal microbes play an important role in the survival and health of wild animals. Spotted hyena (Crocuta crocuta) is one of the representative carnivores in Africa. In this study, we examined the fecal microflora of spotted hyena by conducting high-throughput sequencing of the fecal microbial 16S rRNA gene V3-V4 high mutation region. The effects of age, sex, and feeding environment on the fecal microbiota of spotted hyenas were determined. The results showed that the core bacteria phyla of spotted hyenas fecal microbiota include Firmicutes (at an average relative abundance of 53.93%), Fusobacteria (19.56%), Bacteroidetes (11.40%), Actinobacteria (5.78%), and Proteobacteria (3.26%), etc. Age, gender, and feeding environment all had important effects on the fecal microbiota of spotted hyenas, among which feeding environment might be the most significant. The abundance of the Firmicutes in the adult group was significantly higher than that in the juvenile group, whereas the abundance of Fusobacteria, Bacteroidetes, and Proteobacteria were significantly lower than that in the juvenile group. The abundance of Lachnospiraceae and Ruminococcaceae in the female group was significantly higher than that in the male group. There were significant differences between the fecal microbial communities of Jinan group and Weihai group, and microbes from the phyla Firmicutes and Synergistetes were representative species associated with the difference.