At the end of the shelf life of crayfish stored at 4 ℃, the dominant spoilage organisms in crayfish from Xiaogan, Wuhan were isolated and identified by high-throughput sequencing, and the changes in the bacterial community structure and diversity in crayfish during storage were analyzed. The results showed that 10 spoilage bacterial strains were obtained by the traditional culture-dependent method, including Shewanella baltica, Aeromonas veronii and Proteus vulgaris, etc. Aeromonas sp. and Shewanella sp. were confirmed as the dominant spoilage organisms, with the largest relative abundance at the end of crayfish spoilage. At the same time, the results of high-throughput sequencing showed that the abundance, diversity and evenness of the microbial community decreased with storage time, which was related to the rapid growth of the dominant spoilage organisms and the inhibition of other kinds of microorganisms at the end of the spoilage stage.