A 60-day feeding trial was carried out to determine the effect of dietary lipid levels on growth and physiometabolic responses to optimize the dietary lipid requirement for maximizing the growth of Genetically Improved FarmedTilapia (GIFT) juveniles reared in inland ground saline water (IGSW) of medium salinity (15 ppt). Formulation and preparation of seven heterocaloric (389.56-449.02 Kcal digestible energy/100 g), heterolipidic (40-160 g/kg), and isonitrogenous (410 g/kg crude protein) purified diets were done for conducting the feeding trial. Random distribution of 315 acclimatized fish (mean weight
1.90
±
0.01
g) was made in seven experimental groups such as CL4 (40 g/kg lipid), CL6 (60 g/kg lipid), CL8 (80 g/kg lipid), CL10 (100 g/kg lipid), CL12 (120 g/kg lipid), CP14 (140 g/kg lipid), and CL16 (160 g/kg lipid) with 15 fish per triplicate tank (fish density, 0.21 kg/m3). Respective diets were used for feeding the fish at satiation level three times daily. Results indicated that weight gain percentage (WG%), specific growth rate (SGR), protein efficiency ratio, and protease activity significantly increased up to 100 g lipid/kg fed group, and then the values significantly decreased. Muscle ribonucleic acid (RNA) content and lipase activity were highest in 120 g/kg lipid-fed group. RNA/DNA (deoxyribonucleic acid) and serum high-density lipoproteins levels of 100 g/kg lipid-fed group were significantly higher than 140, and 160 g/kg lipid-fed groups. The lowest feed conversion ratio was found in the 100 g/kg lipid-fed group. The amylase activity was significantly higher in 40 and 60 g lipid/kg fed groups. The whole-body lipid level was increased with increasing the dietary lipid levels, whereas, there was no significant difference in whole-body moisture, crude protein, and crude ash contents of all groups. Highest serum glucose, total protein and albumin, and albumin to globulin ratio and lowest low-density lipoproteins level were found in 140 and 160 g/kg lipid-fed groups. Serum osmolality and osmoregulatory capacity did not vary significantly, whereas carnitine palmitoyltransferase-I and glucose-6-phosphate dehydrogenase showed an increased and decreased trend, respectively, with the increasing dietary lipid levels. According to second-order polynomial regression analysis based on WG% and SGR, the optimum dietary lipid for GIFT juveniles in IGSW of 15 ppt salinity was found to be 99.1 and 100.1 g/kg, respectively.