A 90-day feeding trial was conducted to investigate the effects of fish oil substitution with microalgae meals on growth performance, tissue proximate composition, and biochemical indices in juvenile starry fiounder, (initial body weight 7.35 g±0.03 g). Four groups of isonitrogenous and isolipidic diets were formulated using different primary lipid sources:fish oil (FO), meal (NO), and the mixture of these two algae meals (MO), with the remaining part filled with corn oil. Results showed that growth performance was not significantly different between FO and SO, whereas specific growth rate (SGR), protein efficiency ratio (PER), and viscerosomatic index (VSI) of NO and MO were significantly lower than FO (<0.05). Feed efficiency (FE) of MO and NO was the opposite. The body crude protein content of MO was significantly higher than that in the other groups (<0.05), while body crude lipid content of FO and MO were significantly higher than that in the other groups (<0.05). The muscle ash content of FO was significantly lower than that in the other groups (<0.05). Fatty acid composition of muscle and liver were directly influenced by dietary fatty acid composition ( and DHA in muscle were positively correlated with that of the diets (<0.05), and the percentages of C_14:0 and C_16:1n-7 were significantly positively correlated with that of the diets (18:2n-6, n-6 PUFA, and DHA:EPA in liver were also positively correlated with that of the diets (16:1n-7 showed a significantly positive correlation (<0.01). Compared with FO, levels of serum alanine aminotransferase (ALT), triglyceride (TG), cholesterol (CHO), and alkaline phosphatase (ALP) of NO and MO were significantly decreased (<0.05). Additionally, levels of serum aspartate aminotransferase (AST) and glucose (GLU) of NO were significantly lower than those in other groups ( < 0.05). In summary, there were no negative effects on growth and physiology when fish oil was replaced with P. stellatus to a certain extent.