Experimental diets were formulated containing six levels of linolenic acid (0%, 0.5%, 1.0%, 1.5%, 2.0% and 2.5%) and each diet was fed to darkbarbel catfish Pelteobagrus vachelli in triplicate for 84 days. Weight gain, specific growth rate and feed efficiency increased with the increasing levels of dietary linolenic acid, but were not significantly different among diets containing 1.0%, 1.5%, 2.0% and 2.5% linolenic acid. Red blood cell count and haemoglobin were significantly higher in fish fed the 1.0% linolenic acid than other diets. Lysozyme activity and immunoglobulin M content in fish fed 1.0% of linolenic acid were significantly higher compared with the other diets. Antibody titre in fish fed 1.0-2.5% linolenic acids was significantly higher than in fish fed 0 or 0.5% linolenic acid 14 days after Aeromonas hydrophila challenge. This study indicates that linolenic acid is essential to darkbarbel catfish. Two-straight broken-line analysis with SGR showed that a dietary level of 1.29% linolenic acid is the minimum required for adequate growth of darkbarbel catfish. Considering other measured indicators of fish performance, a level of 1.0-1.5% linolenic acid in the diet is recommended for the darkbarbel catfish.