Arachidonic acid (ARA, 20:4n-6) fed to Seriola dorsalis juveniles at different levels was evaluated. After a seven-week feeding trial, growth performance, hepatopancreas and muscle fatty acid (FA) composition, expression of lipid-relevant genes, and blood parameters were evaluated. Four isoproteic and isolipidic experimental diets were formulated to contain 44% crude protein and 11% lipids with graded inclusion levels of ARA, 0% (Control), 0.4, 0.9, and 1.4% of the total diet. S. dorsalis juveniles (14.54 ± 0.18 g) were randomly divided into twelve tanks with fifteen animals each. The animals were hand fed three times per day to apparent satiation. Dietary treatments did not significantly affect the growth performance, SGR, FCR, and feed intake of fish. Different levels of ARA in the experimental diets directly influenced liver and muscle FA profiles, with significant changes in ARA and EPA deposition between Control treatment and 1.4%, in both tissues. The expression of arachidonate 5-lipoxygenase (alox5), acyl-CoA dehydrogenase very long chain (acadvl), carnitine O-palmitoyltransferase 1(cpt1a) was significantly affected by dietary treatments, with an expression increasing accordingly to the increasing ARA levels. In contrast, a reduction of fatty acid synthase (fas) and proliferator-activated receptor alpha (ppara) expression was significantly reduced as ARA increased in the diet. In addition, a significant reduction in blood cortisol and glucose was found at a 0.9% ARA level compared to the other treatments. Based on the performance, cortisol levels, the gene expression for eicosanoids synthesis, and lipid metabolic pathways, the present study suggests a maximum ARA inclusion of 0.9% in diets for California yellowtail juveniles, S. dorsalis.