Antimicrobial peptides are small, cationic, amphipathic and evolutionarily conserved molecules of the innate immune system of eukaryotes, which they use to wade off a wide range of microbes. The present study was carried out to evaluate longfin snake eel, pisodonophis cancrivorus as a potential source of antimicrobial peptides. The crude peptide extract was obtained by modified acetic acid-acetone precipitation method and its activity was detected by disc diffusion assay. The crude peptide extract was subjected to solid phase extraction using Sep-pak®C-18 cartridges (Waters, USA). The 40% Sep-pak® fractions eluted was further purified by cation exchange chromatography which yielded 5 fractions. Among the five fractions, Pc40-4 and Pc40-5 exhibited the highest antimicrobial activity and showed inhibition against all tested bacterial strains namely Edwardsiella tarda , Bacillus cereus , Vibrio fluvialis , Vibrio cholera , Aeromonas hydrophila and Staphylococcus aureus . Pc40-3 exhibited remarkable antibacterial activity against the Gram-negative bacteria Aeromonas hydrophila (98%). This is the first report on AMPs from Pisodonophis cancrivorus 

 Antimicrobial peptides, Pisodonophis cancrivorus , antimicrobial activity, peptide extract, teleost fish.