The sustainable expansion of aquaculture relies on a sufficient supply of eggs and larvae, which are the first step of life cycle management. However, marine fish larval rearing generally depends on live feed production, which needs additional facilities and labor. The flathead grey mullet (Mugil cephalus), a promising species for aquaculture diversification, has a precocious digestive system development, supporting the feasibility of early weaning strategies. For these reasons, this study evaluated survival, growth, proximate and fatty acid composition, and gene expression of Mugil cephalus larvae reared under three different weaning protocols. Three co-feeding treatments, two with different Artemia sp. concentrations (A100 and A50, 2 and 1 Artemia sp. mL⁻¹ day⁻¹, respectively) and one with only rotifers administered as live feed along the feeding trial (A0), were assessed from 22 to 36 days post-hatching (dph). The A0 treatment performed better in survival (64.79 ± 7.40%) than the A100 protocol (32.46 ± 12.82%). In contrast, the larvae of the A100 treatment presented significantly higher final length (15.51 ± 0.86 mm) than those of the A0 treatment (12.19 ± 1.45 mm) and higher final weight (41.28 ± 1.48 mg) than those of the A50 and A0 treatments (31.23 ± 3.65 mg and 24.03 ± 7.99 mg, respectively). On the other hand, the expression of digestive enzyme- and somatotropic factor-related genes did not show differences between treatments. The present results support the convenience of treatment A0 in maximizing survival, as rotifers should be maintained until 30–32 dph (until a total larval length of at least 10 mm). However, to improve growth and minimize size dispersion, Artemia sp. addition is recommended from day 26 to day 29 post-hatching (total larval length of 8 to 9 mm).

Subadult flathead grey mullets (Mugil cephalus) were fed three experimental diets containing increasing percentages of partially defatted black soldier fly (Hermetia illucens, BSF) that proportionally replaced the protein sources in the reference diet. At the end of the feeding trial, fish growth and gut, liver and spleen histology were evaluated. BSF inclusion did not significantly affect growth performances in any dietary groups. However, Fulton’s condition factor was lower in fish who were fed diets with the highest replacement levels (15% and 20%; BSF15 and BSF20) compared to those fed the 10% replacement (BSF10) and the control diet (BSF0). Histological analyses revealed increased villi thickness and mucous cell proliferation in the intestine of fish from BSF10 and BSF15 groups. A significant worsening of intestinal condition was observed in fish from the BSF20 group. Liver histology was not affected, while a dose-dependent effect on spleen was observed in fish fed BSF inclusions. Despite the absence of enteritis signs, an increase in macrophages/like TNF-α+ cells in the intestine mucosa indicated immune stimulation in the BSF10 group. Results indicate, for the first time, a threshold for BSF meal inclusion in diets specifically formulated for flathead grey mullets. The BSF10 diet was optimal, as growth performance was not affected and intestine health improved at both morphological and innate immune system levels.