Through the single factor test and orthogonal test on the three factors of keeping alive temperature(12, 15, 17, 20, 23, 25 ℃), shrimp to water mass ratio(1∶6, 1∶8, 1∶10, 1∶12, 1∶15) and holding time(0, 12, 24, 36, 48 h), find the optimal conditions for the low temperature keeping alive of Macrobrachium rosenbergii, and detect changes in the water quality indicators and shrimp tissue indicators including muscle, blood, liver, etc, to explore the mechanism of affecting the survival of Macrobrachium rosenbergii. The results showed that the critical temperature of Macrobrachium rosenbergii was 9 ℃; in the single factor experiment, the best keeping alive temperature was 20 ℃, the best shrimp to water mass ratio was 1∶10, and the best holding time was 24 h; in the orthogonal test, the optimal process was keeping alive temperature of 20 ℃, shrimp to water mass ratio 1∶12, holding time 24 h, and the survival rate within 24 h of keeping alive could reach 100%. The optimal process conditions after orthogonal optimization were used for keeping alive test in the low-temperature of Macrobrachium rosenbergii, the concentration of ammonia nitrogen and the total number of colonies in the water samples increased significantly(P＜0.05), but the dissolved oxygen mass concentration and pH were still changing within the suitable range. The mass fraction of muscle glycogen, pH and ATP mass fraction decreased, lactic acid concentration increased significantly(P＜0.05), blood ALT and AST activities increased significantly(P＜0.05), liver glycogen mass fraction decreased significantly(P＜0.05), T-SOD activity first decreased and then increased, while the content of MDA first increased and then decreased. It shows that changes in water quality indicators would affect the normal physiological metabolism of shrimp muscles, blood and liver tissues, thereby affecting their survival time and survival rate.