The invention relates to a controllable temperature water body total dissolved gas supersaturation generation and an experimental device for studying the influence thereof on fish. The device comprises an autoclave, an air inlet, an adjusting air valve, a safe air valve, an air gauge, a water inlet, a water outlet, a water pool provided with a temperature controller, a water pump, a first ball valve, a first flowmeter and a water inlet, wherein, the autoclave, the air inlet, the adjusting air valve, the safe air valve, the air gauge, the water inlet and the water outlet are arranged on the autoclave, and the water pump, the first ball valve, the first flowmeter and the water inlet are sequentially connected with the water pool; an air compressor is connected with the air inlet; the water outlet is sequentially connected with a second ball valve, a second flowmeter and a water channel; a total dissolved gas tester and a water quality monitor are arranged outside the water channel; and the water channel is communicated with the water pool via a latex tube. The invention generates water body featuring total dissolved gas supersaturation by autoclave sealing pressurization, and the whole device adopts circulating water and controls water temperature thereof to be constant. The invention can realize the control on measurement of major physical parameters, such as water temperature,water body pressure, airation content, flow and the like, which affects total dissolved gas supersaturation generation so as to quantificationally analyze the generation principle of total dissolved gas supersaturation and study the influence thereof on fish.