Salmon farming in Chile faces the dual challenge of maintaining growth while improving sustainability. One approach to advance sustainability involves the feeds without marine-fishery inputs. This study aimed to evaluate the effect of completely replacing fish oil (FO) on the performance of Salmo salar throghout marine grow-out. The investigation involved fish ranging from 400 g to 4000 g and explored three diets without fish meal, each which included 2.2% EPA + DHA from different sources: FO (control), transgenic canola oil (Can n-3) and Schizochytrium sp. oil (AOV) as unique type of oils. Each diet was randomly distributed in triplicate among nine tanks with a recirculation system maintained at 12 °C, with automatic feeding and a 24-h light photoperiod. Fish density was 29 fish m−3 at day 133 of the experiment and seven fish m−3 at day 319. The diets were isoproteic, isolipidic and isoenergetic. Fish were evaluated at 133 and 319 days of experiment in terms of productive performance, liver health, EPA + DHA and lipid retention efficiency, fatty acid metabolism, response to acute stress, and final product quality. The results showed similar productive performance and liver health among the three diets during both sampling periods. No differences were observed in lipid or EPA + DHA retention efficiency in the whole body or fillet. Eicosanoids and receptors for anti-inflammatory mediators derived from EPA and DHA showed similar or higher capacities in fish fed with Can n-3 and AOV than those fed FO. Responses to induced stress, measured through lactate, cortisol, sodium, potassium, chloride, glucose, and the liver profile were similar between the three diets. Similarly in fillet quality between the three diets. However, sensory analysis revealed a lower intensity of “global aroma” in Can n-3-fed fish fillets and lower evaluations in “typical salmon aroma”, “oily aroma” and “fresh flavor” in AOV-fed fish fillets when they are compared with FO. The nutritional indicators of the final product were similar between the three diets, except for PUFA/SFA, the thrombogenicity index, hypercholesterolemia/hypocholesterolemia, and the health promotion index. In these specific indicators, fish fed with Can n-3 and AOV demonstrated more favorable results for consumer health than the FO-fed fish.