Aeromonas hydrophila (Ah) is an opportunistic Gram-negative bacterium and a serious global pathogen causing Motile Aeromonas Septicaemia (MAS) in fish and many other vertebrates. This study aimed at identifying the cellular proteins and processes altered by Ah infection in gut tissue of Labeo rohita, an important aquaculture species. Discovery based proteomics data could identify a panel of differentially expressed proteins (DEPs) between Control and AH condition (Ah infected group). A panel of proteins from important pathways were selected for validation of differential abundances using Selected reaction monitoring (SRM) approach.