A 3 × 3 factorial study was conducted to evaluate the effect of three fixed levels of docosahexaenoic acid meal (DHAM) and arachidonic acid meal (ARAM), produced by using a meal that had high levels of the desired fatty acid (0.23% DHAM–0.05% ARAM, 0.50% DHAM–0.10% ARAM, and 0.75% DHAM–0.15% ARAM), and three n‐3/n‐6 dietary ratios (0.3, 0.8, and 1.8), as well as their potential interaction on growth, survival, and fatty acid composition of hepatopancreas and muscle tissue of juvenile Pacific white shrimp, Litopenaeus vannamei, cultured in low salinity. Two additional reference diets with menhaden fish oil or soy and flax oils (n‐3/n‐6 ratios of 1.8 and 1.7, respectively) were tested. No significant differences (at P < 0.05) and no significant interactions were observed among treatments for final weight, weight gain, or survival after 6‐wk feeding. This study confirmed that supplementation of DHA and ARA from alternative sources to fish oil is effective in promoting growth and survival of juvenile L. vannamei. The fatty acid profile and n‐3/n‐6 ratio of shrimp tissue reflected that of dietary lipids, although more studies are required to elucidate how the n‐3 and n‐6 fatty acid balance in the diet relates to shrimp growth.