A 20-week feeding trial was conducted to determine whether increasing linolenic acid (18:3n-3) in vegetable oil (VO) based diets would lead to increased tissue deposition of 22:6n-3 in Nile tilapia (Oreochromis niloticus). Five isonitrogenous and isoenergetic diets were supplemented with 3% of either linseed oil (LO), a mixture of linseed oil with refined palm olein oil (PO) (LO-PO 2:1) and a mixture of refined palm olein oil with linseed oil (PO-LO 3:2) or with fish oil (FO) or corn oil (CO) as controls. The PO-LO, LO-PO and LO diets supplied a similar amount of 18:2n-6 (0.5% of diet by dry weight) and 0.5, 0.7 and 1.1% of 18:3n-3, respectively. Increased dietary 18:3n-3 caused commensurate increases in longer-chain n-3 PUFA and decreases in longer-chain n-6 PUFA in the muscle lipids of tilapia. However, the biosynthetic activities of fish fed the LO-based diets were not sufficient to raise the tissue concentrations of 20:5n-3, 22:5n-3 and 22:6n-3 to those of fish fed FO. The study suggests that tilapia (O. niloticus) has a limited capacity to synthesise 20:5n-3 and 22:6n-3 from dietary 18:3n-3. The replacement of FO in the diet of farmed tilapia with vegetable oils could therefore lower tissue concentrations of 20:5n-3 and 22:6n-3, and consequently produce an aquaculture product of lower lipid nutritional value for the consumer.