This study aimed to investigate the effects of feeding frequency and feeding levels on the physiological responses of Procambarus clarkii，with the goal of optimizing feeding strategies to enhance feed utilization and reduce environmental waste discharge.A total of 810 crayfish，with an average body weight of （4.97±0.11） g were randomly divided into 9 treatment groups，each with 3 replicates，following a 3×3 two-factor experimental design.The daily feeding frequencies were once （F1），twice （F2），and three times （F3），while the feeding levels were set at 3% （L3），4.5% （L4.5），and 6% （L6） of the experimental shrimp’s body weight.The experiment lasted for 56 days.The results showed that the weight gain rate （WGR） and specific growth rate （SGR） of shrimp in the L4.5 and L6 groups were significantly higher than those in the L3 group （P<0.05）.The L6 group exhibited significantly higher WGR，SGR，and feed conversion ratio （FCR） compared to the L3 and L4.5 groups （P<0.05）.In terms of whole-body and muscle nutrient composition，the crude protein content in the L4.5 and L6 groups was significantly higher than that in the L3 group （P<0.05），while the crude fat content in the whole body of the L6 group was significantly higher than in that in the L4.5 and L3 groups.Serum biochemical analysis revealed that levels of alkaline phosphatase （AKP），acid phosphatase （ACP），superoxide dismutase （SOD），catalase （CAT），and malondialdehyde （MDA） in the L6 and L4.5 groups were significantly higher than those in the L3 group （P<0.05）.The MDA level in the F1 group was significantly lower than in the F2 and F3 groups （P<0.05），with significant interaction effects observed between feeding frequency and level （P<0.05）.Based on WGR，survival rate，and feed cost，the L4.5 group achieved the highest economic benefit （approximately 23.45 yuan/kg），followed by the L6 group，while the L3 group yielded the lowest profit （13.65 yuan/kg）.The L4.5F2 group exhibited the lowest feed cost and the highest economic return.Considering growth performance，health status，and cost-effectiveness，a feeding level of 4.5% of body weight，administered twice daily，is recommended for rice-crayfish integrated farming systems.