The invention discloses a method for evaluating the influence of to-be-tested objects (an anti-fatigue drug, an anti-fatigue and a pollutant) on the exercise tolerance of an organic body and special equipment of the method. The method comprises the following steps: (1) correcting the flow speed of a system water body; (2) screening an experimental fish shoal; (3) testing the swimming resistance; and (4) collecting experimental data, and analyzing a result. The special equipment comprises a water tank, a water pump, a container for swimming of fishes, a flow regulation valve, a flow meter, a nitrogen bottle, an oxygen bottle, a refrigerator and a high-speed camera. The method can be used for carrying out pre-warning judgment on an abnormal environmental water body, testing a drug, evaluating the exercise tolerance of different species of small-sized fishes in different water body environments, evaluating and testing the a nutritious supplementary and the like. The equipment can be automatically assembled and occupies a small area, parameters such as the temperature, the water flow speed and the dissolved oxygen amount can be controlled, and the equipment is easy and convenient to operate, relatively low in manufacturing cost and convenient for the application of relevant biological experiments.