Abstract:[Background] Pingdingshan is rich in hot spring resources. The study of microbial diversity and community structure in the spring water will help us to better exploit these resources. [Objectives] The bacterial diversity of Five Hot Spring (SHANGT, ZHONGT, XIAT, WENT and SHENT) in Henan Lushan was investigated and analyzed. [Methods] The bacterial 16S rRNA V3?V4 genes were sequenced by Illumina Hiseq2500 PE250 technology. The OTUs was sorted and counted using Uparse, Mothur, Silva, MUSCLE and Qiime software. Finally, the bacterial abundance and diversity in Five Hot Spring was analyzed. [Results] It was obtained 56 017, 68 319, 65 247, 59 340, 68 825 effective tags, and acquired 670, 287, 337, 598, 381 OTUs from the SHANGT, ZHONGT, XIAT WENT, and SHENT respectively. The analysis of bacterial classification showed that the bacteria from Five Hot Springs could be classified as more than 40 phyla. Proteobacteria accounted for 84.12% was the dominant bacteria. In the genera level, bacteria from Five Hot Springs could be classified into more than 239 genera. The dominant bacteria of Five Hot Springs (SHANGT, ZHONGT, XIAT, WENT and SHENT) respectively is Acinetobacter, Tepidimonas, Vogesella and Acinetobacter, Sphingopyxis and Novosphingobium, Aquabacterium. [Conclusion] Five Hot Springs in Lushan are rich in microbial resources and may have the potential for the development of such resources.