Abstract: The effects of holding time,temperature,density,salinity and cooling model on the survival time of grouper(♀Epinephelus fuscoguttatus×♂Epinephelus lanceolatus)were studied by single factor experiment and the orthogonal experiment. The water quality index and serum biochemical index were analyzed in order to explore the optimum conditions for the conservation of grouper(♀E. fuscoguttatus×♂E.lanceolatus)with water. Results showed that the survival time of the grouper was(168±8)h after holding for 48 hours,then inducing to dormant by a gradient cooling model at the average reducing ratio of 1~2℃/h and keeping alive under the condition of 15℃,25‰ salinity and 100 g/L of fish density. The total ammonia nitrogen content in water after keeping alive for 72 h was(7.28±0.06) mg/L. Blood glucose,globulin concentration and serum alanine aminotransferase activity were increased significantly(p<0.05),while cortisol,triglyceride,urea nitrogen concentration and aspartate aminotransferase and lactate dehydrogenase activities were not changed significantly. The results showed that this survival condition is suitable for the long distance transportation of grouper(♀E. fuscoguttatus×♂E. lanceolatus).