The oriental fruit fly, Bactrocera dorsalis (Hendel), is one of the most destructive agricultural pests and hosts diverse intestinal bacterial communities. We used 16S rRNA gene sequencing to investigate the microbial communities associated with the intestines of females and males from two B. dorsalis populations collected from Hainan and Guizhou Provinces of China. A total of 260,615 high-quality 16S rRNA gene reads with an average length of 253 bp were obtained. Highly diverse bacterial communities were observed across individuals, with communities containing between 691 and 1,262 bacterial operational taxonomic units. In addition, 37 bacterial phyla, 65 classes, 130 orders, 198 families, 201 genera, and 70 annotated species were identified, wherein the Proteobacteria were the most dominant phylum among all of the libraries, followed by the Firmicutes and Bacteroidetes. Bacterial community composition varied by host sex and geographic location of the populations. Female gut communities comprised fewer bacterial families than males, and females had lower relative abundances of 33 of the 35 most abundant families. In addition, female gut communities harbored greater abundances of Enterobacteriaceae than males. The Hainan population gut communities contained fewer bacterial families than those of the Guizhou populations. However, 5 of the 35 most abundant families were more abundant in communities from the Hainan population than those of the Guizhou population. The results of this study help us better understand the importance of symbiotic bacteria in B. dorsalis and provide a foundation for assessing the use of gut microorganisms as bio-control agents for these serious pests.