Seahorses are increasingly regarded as a promising farming object suitable for recirculating aquaculture systems (RASs) due to their high economic value. However, reports on the large-scale farming of seahorses in RASs are rare, and some key parameters, such as stocking densities, are still unclear. In the present study, we employed the lined seahorse (Hippocampus erectus), for which large-scale farming has been achieved, to determine the suitable stocking density for three different-sized juveniles in RASs. The three different-sized juveniles had body heights of 4.0, 7.0, and 9.0 cm, and their test density gradients were 1.0, 0.8, 0.6, and 0.4 inds/L; 0.6, 0.5, 0.4, and 0.3 inds/L; and 0.4, 0.3, 0.2, and 0.1 inds/L, respectively. The juveniles were cultivated for one month, and then their survival, growth, and plasma cortisol and brain serotonin contents (two stress-related indicators) were analyzed. The results show that, regardless of the size of the juveniles, a high density can inhibit growth and trigger stress responses. In addition, for small- (4.0 cm) and medium-sized (7.0 cm) juveniles, a high density can also exacerbate size heterogeneity and cause death. Taking into account the welfare and yield of farmed seahorses, the present study suggests that the suitable stocking densities for 4.0, 7.0, and 9.0 cm juveniles in RASs are 0.6, 0.4, and 0.2 inds/L, respectively.