Vibrio parahaemolyticus is a zoonotic pathogen that poses a serious threat to animal, fish and human health. In order to study the dynamic process of host neutrophils and bacteria infection, we established a model of local infection of the 48 hours post fertilization (hpf) zebrafish larvae otic vesicle infected with the V. parahaemolyticus strain labelled with red florescent protein (Vp57^RFP) in this study, and the Escherichia coli Ec01 strain was used as a control to systematically compare the median lethal dose (LD₅₀) and survival curves, infection process and dynamic process of interaction with neutrophils of V.parahaemolyticus and E. coli, respectively. RT-qPCR verified the expression levels of the inflammation-related genes il1b and il10. The results showed that the LD₅₀ of E. coli and V.parahaemolyticus infected zebrafish larvae were 1.14×10¹¹ CFU/mL and 7.90×10⁷ CFU/mL, respectively. Compared to E. coli, V. parahaemolyticus recruited faster and more numerous neutrophils during dynamic infection imaging, and induced more il1b and il10 genes expression level that caused strong inflammatory response in zebra fish larvae at the same time point. Based on the above research, we successfully established the model of V.parahaemolyticus-otic vesicle zebrafish larvae infection, which provides a basis for further study of the dynamic process of neutrophil clearance of Vibrio parahaemolyticus infection and the in-depth revealing of the innate immune response mechanism.