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The double-row and double-chamfered aquaculture tank is a special tank structure of the aquaculture ship. The tank sloshing of this structure is coupled with the hull motion, which has an important impact on the safety of the hull motion. In the present study, research on the tank sloshing and hull motion response of aquaculture ships was conducted based on the model seakeeping and tank sloshing tests in regular waves. The test results were compared with the numerical simulation results of solid loading without sloshing. The results showed that the numerical simulation of the pitch motion was consistent with the amplitude-frequency response curve of the experimental results. Under certain transverse wave conditions, a large discrepancy existed between the amplitude-frequency response curve of the heave motion by the numerical simulation and the test results, and the roll motion differed most from the experimental result. Severe roll resonance occurred when the wave length-ship length ratio was 0.6. The roll motion amplitude was increased by 183.2%. Therefore, compared with aquaculture ships without sloshing, the sloshing of the tank has little effect on the pitch but has a great impact on the roll and heave motions, with the most significant effect on the roll motion.