The feeding trials were carried out with freshwater crayfish (Procambarus clarkii) to evaluate the three feed stimulants betaine (Bet), squid extract (SE) and squid meal (SM) upon fish meal (FM) diets or biofeed (BF) diets in which FM were partially replaced by BF. Results showed the three feed stimulants had no significant effect on growth. By the relative concentration of markers in the faeces and diets, the preference for each diet was estimated, and the best stimulant was the mixture of Bet, SE and SM. As FM replaced by BF increased, added with the mixture, the relative weight gain showed an increasing trend, but decreased significantly by 30% replacement. With the substitution increasing, trypsin activities increased, whereas decreased too upon 30% substitution. Less than 30% replacement, there was no significant effect on muscle proximate composition, intestine amylase activities and serum haemocyanin contents, except the drop of lipid. The results indicated that the mixture of Bet, SE and SM could perform well as a feed stimulant. And with the addition of the mixture, FM in the diet could make about no more than 20% saving without any adverse effects. 3 Biofeed (g kg À1 ): crude protein 437, crude lipid 58, calcium 17, phosphorus 16.7, moisture 120 and ash 170. 4 Other components (g kg À1 ): rapeseed meal 100, soybean meal 93, vinasse 80, cottonseed meal 40, corn protein 60, flour 180, soybean oil 18, fish oil 18, mineral mixture 5, vitamin mixture 5, calcium dihydrogen phosphate 15, compound phosphatide 20, moult hormone 1, choline 1, dextrin 10, gelatine 40 and markers: diets of the same group (fish meal or biofeed) were marked with 100 mg kg À1 of the oxides of rare earth element, respectively (