The shelf life and survival bacterial species of ready-to-eat Penaeus vannamei during ambient temperature storage were studied. The quality changes and shelf life were observed by the sensory, physico-chemical, microbiological evaluation and electronic nose analysis, the bacteria species were identified by 16S rRNA sequencing. The results showed that initial sensory quality, physical and chemical indicators was pretty good. In the first 4 months, the water content, water activity, pH, sanility and color (red degree and yellow degree) had no significant differences, and then, the water content and Aw decreased, but pH and sanility increased, while the red degree and yellow degree fluctuated; The chewiness, hardness and elasticity had no siginficant differece in the first 3 months, and then increased firstly and decreased secondly, which kept fluctuating in cohesive force in the storage. TVBN and TBA did not exceed the limited value during the 9 months ambient temperature storage, with the total colony lower than 1 000 CFU/g; Bacillus licheniformis and Bacillus subtilis were found to be residue in individual expansion bags. In conclusion, through previous criteria of sensory, physicochemical and microbiological evaluation, the shelf life of Penaeus vannamei was 9 months.