In order to study the effects of different feeding frequencies and feeding levels on the growth, muscle nutrients and immune function of Procambarus clarkii, 3×3 two-factor design was used. The daily feeding frequencies were one (F1), two (F2) and three (F3) times, and the feeding levels were 3 % (L3), 4.5 % (L4.5) and 6 % (L6) of the experimental shrimp fresh weight. A total of 9 experimental groups were fed with (4.97±0.11) g crayfish for 56 days. The results showed that the weight gain rate and specific growth rate of shrimp in L4.5 and L6 feeding levels were significantly higher than those in L3 experimental group (P<0.05). The weight gain rate and specific growth rate of shrimp in L6 group were the highest, and the feed ratio of shrimp in L6 level was significantly higher than that in L3 and L4 (P<0.05). In terms of conventional nutritional composition of whole shrimp and conventional nutritional composition of muscle, the crude protein content of L4.5 and L6 experimental groups was significantly higher than that of L3 group (P<0.05), and the crude fat content of L6 was significantly higher than that of L4.5 and L3 experimental groups. In terms of serum biochemical indexes, the serum AKP, ACP, SOD, CAT and MDA levels of shrimp at L6 and L4.5 feeding levels were significantly higher than those of L3 group (P<0.05), and these indexes in F1 experimental group were significantly lower than those in F2 and F3 experimental groups, and there was a significant interaction between the two experimental factors (P<0.05). Combined with weight gain rate, survival rate and feed cost, the breeding benefit of L4.5 experimental group was significantly higher than that of other experimental groups (P<0.05) about 23.45 yuan /kg, followed by L6 and L3, which was the lowest about 13.65 yuan /kg. The feed cost of L4.5F2 group was the lowest and the breeding benefit was the highest. Considering the growth and health of the experimental shrimp, as well as the cost and benefit of breeding, the optimal feeding level of 4.5% and feeding frequency of 2 times per day were considered.