AbstractAniline (C6H5NH2) an important intermediate in the organic and fine chemical industry, is ubiquitously used worldwide. It is one of the important building block for manufacturing of 4,4-methylene diphenyl diisocyanate (MDI), accelerators in rubber processing, dyes, tattoo inks, photographic chemicals, antioxidants, corrosion inhibitors, pharmaceuticals and antiseptics. The current study evaluated 96 h LC50 of aniline and based on this, two sublethal concentrations (4.19 mg/l and 8.39 mg/l) were selected for acute exposure studies in freshwater food fish Channa punctatus. Erythrocytes of fish are nucleated hence they play an important role in physiology, immune system, protein signalling and haemostatic condition along with respiration. Blood samples were collected after 24, 48, 72, and 96 h of exposure to study haematological, cytotoxic and genotoxic effects of sublethal concentrations of aniline in C. punctatus. Symbolic elevation in time and dose dependent DNA damage was observed by comet assay as well as micronuclei assay revealing maximum damage after 96 h of exposure. After aniline exposure, scanning electron microscopy and ATR-FTIR studies showed anomalies in structure and alterations in biomolecules of RBCs of aniline exposed group as compared to control group respectively. Semi prep HPLC studies revealed bioaccumulation potential of aniline in higher concentration exposed group.