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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).

Mullets (Mugilidae) present significant potential for sustainable aquaculture diversification due to their eurythermal, euryhaline, and low-trophic nature. However, the physiological differences and optimal cultured conditions among the diverse mullet species are quite unknown. For these reasons, the present study aimed to address two main objectives: (1) to characterize the differences in digestive biochemistry, somatic indexes, and body composition between two mullet species (Liza aurata and Chelon labrosus); and (2) to evaluate the interactions of two different feeding frequencies (one against three meals per day) on the above-mentioned parameters, and also on the potential bioavailability of nutrients determined using in vitro assays. The results evidenced higher protease and amylase activities for Chelon labrosus than for Liza aurata, while the latter species presented a higher percentage of eviscerated weight and muscle lipids. Furthermore, the results from in vitro assays supported the higher enzyme activity of Chelon labrosus by an observed increase in the release of amino acids and reducing sugars measured for this species. Regarding feeding patterns, the results of the in vitro assays simulating enzyme: substrate (E:S) ratios corresponding to one or three meals per day point to a clear increase of nutrient bioavailability when the daily ration is split into several meals. The present results improve the physiological knowledge of mullet species and define criteria to develop better management protocols by producers.