An 8‐week feeding experiment was conducted to determine the effect of dietary betaine levels on the growth performance and hepatic intermediary metabolism of genetically improved farmed tilapia (GIFT) strain of Nile tilapia Oreochromis niloticus (mean initial body weight: 78.3 ± 1.3 g, means ± SD). Six practical diets were formulated with the incorporation of betaine at the levels of 0 (control), 5, 10, 15, 20 and 25 g kg−1. Survival showed no significant differences among the treatments (P > 0.05). The highest and lowest weight gain (WG) and specific growth rate (SGR) were observed for fish fed the diets containing 5 and 0 g kg−1 (control) betaine, respectively. Feed intake showed similar trend with WG and SGR. In contrast, feed conversion ratio was the lowest when dietary betaine level was 5 g kg−1. In general, dietary betaine supplementation showed no significant effect on hepatic composition of tilapia. Condition factor and viscerosomatic index tended to increase with increasing dietary betaine levels from 0 to 5 g kg−1 and then decline when dietary betaine levels further increased from 5 to 25 g kg−1. In contrast, hepatosomatic index declined with increasing dietary betaine levels (P < 0.05). Dietary betaine levels significantly influenced several hepatic enzymatic activities, including succinate dehydrogenase, lactate dehydrogenase, malic dehydrogenase, lipoprotein lipase and hepatic lipase, suggesting that dietary betaine addition had significant effects on nutrient metabolism in the liver. Based on the second‐order polynomial regression analysis of WG, 12.5 g kg−1 of dietary betaine level seemed optimal for genetically improved farmed tilapia strain of O. niloticus.