By evaluating the changes of antioxidant enzyme activity, antioxidant capacity, metabolites, glycogen and lactic acid content in tissues, the effects of different packaging oxygenation modes and transport road conditions on oxidative stress of grouper during waterborne transport were studied. The results showed that the antioxidant enzyme activity, antioxidant capacity and metabolites of grouper under continuous aeration and low oxygen vibration changed non-significantly, which indicated that the body could regulate and maintain the balance of antioxidant system. However, the activities of antioxidant enzymes, antioxidant capacity, lipid metabolites and anaerobic metabolic material production in grouper under very low dissolved oxygen and continuous aeration， and the strong vibration environment changed significantly with the prolongation of transport time. It indicated that very low dissolved oxygen content and the severe vibration could lead to a sharp increase in the content of free radicals in the body, which couldn’t be cleared in time, and the oxidative stress reaction was intense, destroying the balance of antioxidant system. It was concluded that different packaging oxygenation modes needed to be designed to keep the suitable alive rate, according to the distance and conditions of the transportation routing.