The invention discloses an additive premix for stereo cultivation of Tilapia mossambica as well as a preparation method and a use method thereof. The premix comprises a material A which is prepared from vitamin A, vitamin D3, vitamin E, vitamin K3, vitamin B1, vitamin B2, vitamin B6, vitamin B16, DL-calcium pantothenate, niacin, folic acid, inositol, 2% biotin, 35% vitamin C, an antioxidant, bacillus subtillis, a mildew preventive, immunopolysaccharide, bile acid and defatted rice bran; the premix which is prepared from the following components such as vital cobalt, copper sulfate pentahydrate, ferrous sulfate, magnesium sulfate, manganese sulfate, zinc sulfate, sodium selenite, choline chloride, an acidifier, calcium dihydrogenphosphate, table salt, garlicin, kelp powder, zeolite powder and the like is added into Tilapia mossambica feed in proportion. Various special components are added to satisfy the demand of Tilapia mossambica on components of vitamins or mineral elements under the stereo cultivation condition, so that the immunity of Tilapia mossambica can be effectively improved and the incidence of Tilapia mossambica under severe environments can be reduced.