Abstract: In order to investigate the effects of underwater noise on behavior (swimming rate, feeding rate, spatial distribution) and gut microorganisms of sturgeon, hybridized sturgeon were exposed to noise (145±5) dB, 400 Hz for 0, 48h, 7d, 7d, and 14d, followed by 48h of recovery in an environment without stressful noise. The results showed that the swimming and feeding rates of hybrid sturgeon were significantly reduced and the spatial distribution was changed after noise stimulation. At the beginning of the noise stimulation, the hybrid sturgeon gathered on the side away from the noise source, but gradually approached the noise source after 3min. The microbial composition was significantly different from the other groups at 7d of noise stress. There was no significant effect of noise on the abundance of gut microorganisms in hybrid sturgeon. Differences in the dominant populations of gut microorganisms existed in each group at different time periods and the dominant populations changed over time. The results of the above microbiological analysis showed that “cell signaling processes” “carbohydrate transport” and “amino acid transport and metabolic functions” were significantly lower than those of the other groups at 48h and 7d of noise stress as predicted by COG. The results showed that noise had significant effects on the feeding rate, swimming rate, and spatial distribution of hybrid sturgeon, changing the composition and proportion of its gut microorganisms and affecting various vital life pathways, such as amino acid metabolism. This experiment simulated the mixed noise of various underwater noise sources and explored their effects on the behavior and gut microorganisms of hybrid sturgeon for the first time, which can provide basic information for the in-depth exploration of healthy ecological breeding and physiological response mechanisms to adversity of hybrid sturgeon.