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As a marine carnivore exhibiting exceptionally high growth rates, cobia are considered a species for which fish oil (FO) replacement may be difficult. However, partial, if not complete, FO replacement is necessary to ensure sustainability. We evaluated the effects of graded substitution of dietary FO with soybean oil (SO) in cobia culture. Feeds contained FO (100% FO), SO (0% FO) or blends of the two (67% FO, 33% FO) as the supplemental lipid source. Production performance was largely unaffected by partial replacement of FO with SO: feed intake and final weight were reduced only in the 0% FO dietary treatment. Fillet total lipid fatty acid (FA) composition differed among the dietary treatments, closely approximating dietary FA profile. As increasing amounts of FO were replaced, SO‐associated FA became enriched within the fillet lipid at the expense of FO‐associated FA. Fillet lipid classes were associated with a particular FA signature, regardless of dietary FA profile. SO can replace a substantial amount of dietary FO; however, juvenile cobia appear to exhibit a nominal requirement for intact long‐chain polyunsaturated FA. Therefore, aggressive FO replacement may result in essential fatty acid deficiencies unless the feeds can be amended with alternative sources of these essential nutrients.

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