Astaxanthin is a valuable pigment source for many aquacultured species, including salmonoids, shrimp, sea bream, and ornamental species. Conjugated linoleic acid (CLA) and ascidian tunic extracts were mixed with the basal diet of rainbow trout to investigate their pigmentation effects. Synthetic Carophyll Pink and natural carotenoids that came from the tunic extracts were incorporated into muscle and skin tissues. The main carotenoids found in muscle after 8 weeks were canthaxanthin in CP12 (13.4%), and CP52 (17.2%), and astaxanthin in CP12 (58.5%), and CP52 (59.2%) in the Carophyll Pink group, while those in skin were canthaxanthin in CP14 (34.5%), and CP54 (29.2%), and astaxanthin in CP14 (32.0%), and CP54 (36.5%) in the ascidian tunic extract group. The total carotenoid content in skin (53.0-69.3 mg/kg) was greater than that in muscle (9.5-13.8 mg/kg).