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To assess the effect of addition of probiotics and carbon source on the microbial community structure within the shrimp culture, an enclosure system simulating a high density, intensive aquaculture of Litopenaeus vannamei was set up in Dianbai, Maoming, Guangdong province, China. The phylogenetic profiles of microbial communities in the enclosure system after addition of Bacillus and molasses were studied in the middle and final culture season (on the 45th, 75th and 95th day) using polymerase chain reaction-denaturing gradient gel electrophoresis. A total of four groups, each containing a triplicate were set up: Group A (control), Group B (64% of daily feed was provided as molasses with Bacillus sp.), Group C (50% of daily feed amount provided as molasses with Bacillus sp.) and Group D (64% of daily feed amount as molasses alone). The results showed that a-Proteobacteria, d-Proteobacteria, Bacteroidetes and Actinobacteria were the dominant bacteria. Moreover, the relative proportion of a-Proteobacteria increased and that of Bacteroidetes decreased gradually with time. Pathogenic Flexibacter was the dominant genus in Group A on the 45th day, while the functional strains (Sphingomonas sp., Erythrobacter sp., Roseobacter sp., and Rhodobacter sp.) were detected multiple times in Groups B and C. Bacillus sp. was the cultivable, dominant bacterium in Groups B and C on the 95th day. Therefore, the combined use of Bacillus and molasses helped increase diversity of the microbial community, effectively inhibit pathogens, and promote the formation and development of a beneficial microbial community structure in the high density, intensive aquaculture of L. vannamei.