Vibrio parahaemolyticus is a foodborne pathogen commonly associated with seafood worldwide. Ethanol is widely used as a disinfectant and preservative in the food industry. Low concentrations of ethanol can inhibit the growth of V. parahaemolyticus and alter its protein expression profile and stress resistance. However, it remains unknown how ethanol exposure affects the behavior and gene expression of V. parahaemolyticus. In this study, RNA sequencing revealed differential expression of 1020 genes in response to 1.5 % (v/v) ethanol, including biofilm-associated genes, c-di-GMP metabolism-related genes, major virulence genes, putative regulatory genes, and Hsp-like protein encoding genes. Specifically, genes involved in exopolysaccharides, type VI secretion system 1, thermostable direct hemolysin, and type III secretion system 2 were upregulated upon exposure to 1.5 % ethanol. Additionally, the data also suggest a significant decrease in swimming and swarming motility, as well as a notable increase in biofilm formation, under 1.5 % ethanol stress. This study enhances our understanding of how V. parahaemolyticus adapts its behavior and gene expression to low concentrations of ethanol.