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A 90-day study was performed under three different stocking densities, including high density (10,000 fishes/cage), medium density (8000 fishes/cage), and low density (6000 fishes/cage), in a deep-sea net cage for yellowtail kingfish (Seriola lalandi). The physiological characteristics and growth performance were tested, and structural characteristics of the gastrointestinal microbiota were systematically analyzed. The results show that fishes with high density had a lower weight gain rate and a specific growth rate, as well as higher serum cortisol content. The diversity, types and numbers of dominant microbiota with significant differences, and the numbers of shared genera among the different groups all changed. Core genera in the gastrointestinal tract were obtained according to the principles of dominance, commonality, and difference. The changes in the relative abundance of the core genera might be related to the growth and physiological characteristics of the host. The ratio of Firmicutes to Bacteroidetes in the stomach and pyloric caecum, which favors the accumulation of energy by the host from the diet, was higher in the medium-density group than in the other groups. This indicates that the higher density could cause physiological stress and affect growth performance. In order to reduce the resulting growth differences, gastrointestinal microbiota might assist the host in accumulating energy, participating in the energy distribution by adjusting its structure. Based on the growth, physiology, and production practices, the medium density was the appropriate density in this study. This study provides a reference for the improvement of deep-sea culture technology and the promotion of healthy growth through the gastrointestinal microecological regulation of yellowtail kingfish. [ABSTRACT FROM AUTHOR]