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The main challenge of the aquaculture relies on the shift from fishmeal to more sustainable ingredients. Interest on carbohydrates has growing rapidly as they are considered as promising resources, providing energy and preserving proteins for growth. However, rainbow trout is considered as a poor user of carbohydrates displaying postprandial hyperglycaemia and impaired growth performances when fed with diets containing more than 20% of carbohydrates. Nonetheless, recent evidence points that broodstocks could be better user of high carbohydrate diet compared to juveniles. However, no study investigates how sex-reversed females – neomales – eat, grow and reproduce under a high carbohydrate diet. Our objective was thus to assess growth and reproductive performances of neomales fed with a high carbohydrate diet during an entire reproductive cycle. Our results demonstrate that neomales display specific metabolic and physiological changes when fed with a high carbohydrate diet compared to both females and males broodstocks as well as compared to juveniles. Altogether, our data demonstrate the critical relevance to formulate specific diets in accordance with specificities of each type of broodstocks (i.e. females, males and neomales).

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Simple Summary Overall, this study has reported that lower stocking density significantly altered survival with several fish dying during the experiment and an alteration of growth and feed efficiency for the remaining fish. Concomitantly, our results showed that low stocking density induced a chronic stress altering the physiological responses of trout (parameters related to welfare, immune and inflammatory systems). Our results supported the hypothesis that a minimum number of fish is essential for culturing fish farmed in order to maintain healthy physiological responses allowing them an optimal growth. Abstract The present study investigated the effect of low stocking density on growth, survival, feed parameters and physiological responses (blood metabolites, welfare indicators, immune biomarkers, and transcriptomic responses of stress and immune-related genes) on juvenile rainbow trout (Oncorhynchus mykiss) reared under a recirculating aquaculture system during 12 weeks. Fish (average weight 29.64 g) were reared in triplicate under four initial densities: nine fish per tank (D9, 3.76 ± 0.06 kg/m3), 18 fish per tank (D18, 7.66 ± 0.18 kg/m3), 27 fish per tank (D27, 9.67 ± 0.01 kg/m3) and 36 fish per tank (D36, 12.94 ± 0.14 kg/m3). Results showed that lower stocking density D9 significantly altered survival with several fish dying during the experiment and an alteration of growth and feed efficiency for the remaining fish. In parallel, the study revealed that low stocking density induced a chronic stress altering the physiological responses of trout by dysregulation of the inflammatory, immune system, and indolamine/catecholamine brain levels. In conclusion, regarding all the variables observed, low stocking density (D9) alters survival, growth and feed efficiency of rainbow trout with alteration of their physiological responses. Selecting appropriate fish density relating to rearing conditions proved to be an essential concern to improve welfare in an aquaculture context.