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A six-week feeding trial was carried out to evaluate the effects of different dietary Myo -inositol (MI) levels on growth, feed efficiency, immunity, and antioxidant properties of juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) fed low-fishmeal diets. Seven experimental diets were made to contain 50, 100, 169, 241, 309, 386, and 459 mg/kg of MI (analyzed values, dry matter basis), being abbreviated as MI-50, MI-100, MI-169, MI-241, MI-309, MI-386, and MI-459, and each MI treatment had three replicates. Twelve fish (average initial body weight: 18.7 ± 0.1 g) were allocated to each replicate, being hand-fed their diets to apparent satiation on 8:00 am and 16:30 pm. The results indicated that within the range of 50–241 mg/kg dietary inositol, weight gain percentage (WG%), feed efficiency (FE) and protein efficiency ratio (PER) were increased with the increasing of MI, and above the 241 mg/kg, they were dropped, especially at 386 and 459 mg/kg. According to the broken-line regression analysis conducted to assess the relationship between WG% and dietary MI levels, the optimum MI dose was 267.9 mg/kg (dry weight) in the low-fishmeal diet of hybrid grouper. Condition factor (CF), viscerosomatic index (VSI), hepatosomatic index (HSI), and intestinal index (ISI) were significantly (P < 0.05) affected by dietary MI levels, varying with a similar trend as WG%. Fish fed MI-241 had the highest values of serum lysozyme (LZM), hepatic total antioxidant capacity (T-AOC) as well as the relative mRNA level of hepatic insulin-like growth factor-Ⅰ (IGF-Ⅰ) among all treatments. In conclusion, optimum dietary MI requirement for the maximal growth of hybrid grouper was 267.9 mg/kg, and suitable dietary MI supplementations improve growth, immunity, and antioxidant properties of juvenile hybrid grouper fed low-fishmeal diets. • An optimal MI content of 267.9 mg/kg is recommended in the low-fishmeal diets of hybrid grouper. • Suitable MI supplementations improve growth, immunity and antioxidant properties of fish. • Dietary MI levels exceeded 368 mg/kg have a negative impact on fish. [ABSTRACT FROM AUTHOR]