Gaotian, one typical conservative village in rural area of South China, is differentiated from other adjacent village for its longevity and health situation of residents. To ascertain the difference of intestinal microbial community between Gaotian and other region, high-throughput sequencing and systematical bioinformation analyses was adopted to compare 21 samples in long life group with 28 in control group. The α diversity showed that the diversity of species of intestinal flora of Gaotian villagers was higher than that of control group, while the β diversity showed that the similarity of intestinal flora for Gaotian residents was also much higher than that of control group. OTU cluster analysis and Venn diagram showed that the intestinal microbial community of Gaotian villagers is different from that of control group. To quantitatively compare the main flora constitution in all samples, real-time PCR was performed, and the results showed that the biomass of Enterococcus, Lactobacillus, Enterobacteriaceae, Clostridium perfringens, and Bacteroides of Gaotian villages is generally significantly higher than that of control group. Remarkably, some special species, i.e., Methanobacterium, Butyricimonas, Deinococcus, and Streptococcaceae, have been found in Gaotian villagers. Overall, this study lays a preparatory basis for exploration of the resources of special species from healthy and long-living elderly Gaotian villagers and for proposal of a hypothesis, namely, the diversity in intestinal flora of Gaotian might contribute to the longevity and health of local residents. Further study should be focused on screening and functional evaluation of the special species in the long-life residents.