The potential immunostimulatory effects of small peptides on sea cucumber, Apostichopus japonicus, were investigated in vivo and in vitro. Small peptides were injected into sea cucumbers and a control group was injected with equal volume of sterile filtered seawater. The nonspecific humoral and cellular responses were determined, respectively, and sea cucumbers were challenged by Vibrio splendidus. The results showed that test groups significantly enhanced most of the immune parameters tested (P < 0.05). Both phagocytic capacity and respiratory burst activity were up to maximum value on the fourth day and the values were 2.3 and 1.4 times as high as the control group, respectively. However, small peptides had no significant effect on total coelomocytes counts. Among the humoral responses, the activities of acid phosphatase, alkaline phosphatase, and superoxide dismutase were up to maximum value on the fourth, fourthth and seventh day, respectively. The values were 2.3, 2.2, and 2.0 times as high as the control group, respectively. At the end of the challenge, the cumulative mortality rate of sea cucumber injected with 0.5 mg small peptides (43.3%) was significantly lower than the control group (66.7%). In conclusion, small peptides as an additive could improve the nonspecific immunity level of Apostichopus japonicus.