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.

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