Abstract: Endogenous spoilage bacteria of Litopenaeus vannamei are the bacterial floras living in Litopenaeus vannamei body which cause the Litopenaeus vannamei spoilage.It is one of the most important factors which cause spoilage while the Litopenaeus vannamei are stored up and in transportation process.In order to find out the endogenous dominant spoilage bacteria of Litopenaeus vannamei under refrigeration,the bacteria separation is applied in experiment and therefore the endogenous spoilage bacteria are gained,and then the bacterial strains of high capability of decomposing protein were obtained after protein comparison experiment.Once more,auto-biochemical identification system and 16S rRNA sequencing are applied in experiment process.The result shows that the endogenous dominant spoilage bacteria of Litopenaeus vannamei are of Brevibacterium, and the semblance of auto-biochemical identification and 16S rRNA sequencing is 98.833% and 99.065% respectively.Researches show that the endogenous dominant spoilage bacteria of Litopenaeus vannamei under refrigeration are of Brevibacterium,and it is possible to research on endogenous freshness preservation technology of Litopenaeus vannamei by controlling Brevibacterium bacteria.