We evaluated the effects of substrate type [no substrate (control), concrete, sand, sand and mud, and gravel] on survival, growth, and behavior of Fenneropenaeus chinensis under laboratory conditions in a semi-intensive culture mode (SICM) and intensive culture mode (ICM). In the SICM, substrate type had no effect on survival (S) and molting frequency (MF) (P>0.05), though S and MF were lowest in the control and highest in the sand and mud substrate treatment (100% and 12.9 %/d, respectively) in the ICM. Substrate type significantly affected the specific growth rate (SGR) of F. chinensis reared in SICM and ICM (P<0.05). In the SICM, the SGR was highest in the sand and mud substrate treatment, and was 1.39 times higher than the SGR in the control. In the ICM, the SGR (1.94 %/d) was highest in the control, but there was no significant difference between the control, sand, and sand and mud substrate treatments (P>0.05). Substrate type had a significant effect on the behavior of F. chinensis (P<0.05). In the SICM, the feeding ratio (FR) and locomotion rate (LR) were all highest in the sand and mud substrate treatment before and after feeding, however, attacking frequency (AF) was highest in the sand substrate treatment before feeding and in the control treatment after feeding, peaking at 4.53 times/(shrimpּh−1). There was no significant difference in AF between the control, sand and mud substrate, and gravel substrate treatments (P>0.05). In the ICM, the FR and AF were highest in the sand substrate treatment before feeding and there was no difference in LR among the substrate treatment groups (P>0.05). The FR was highest in the control and lowest in the gravel substrate treatment after feeding. LR and AF were highest in the sand and mud substrate treatment. Our results suggest that the addition of substrate enhanced the growth and survival of F. chinensis in culture. F. chinensis exhibited behavioral differences when reared over different substrate types and preferred the sand and mud substrate.