Exposure to psychosocial stress is a risk factor for human diseases such as depression. Social defeat stress (SDS) is a well-known rodent model of human psychosocial stress, and animals exposed to SDS show social avoidance behavior. Fish oil, which is rich in docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), is expected to decrease the risk of depressive disorders. In this study, we determined whether fish oil affects the social behavior of SDS-exposed mice and measured serotonin levels and expression of genes related to tryptophan (TRP) metabolism in the hippocampus. The experimental animals were fed a diet containing fish oil during SDS exposure. For the fish oil treatment, experimental mice were fed a diet containing fish oil at low (L-FO), middle (M-FO), and high (H-FO) concentrations. The control group was supplemented with an equivalent amount of canola oil (no fish oil: N-FO). After the SDS protocol, we performed a social interaction test and assessed the sociality of experimental mice. In the N-FO group, SDS-exposed mice showed negative social interactions compared with non-stressed mice. The L-FO and H-FO groups showed negative social interactions after SDS exposure; however, the M-FO group did not exhibit negative social behavior. The serotonin levels of SDS-exposed mice were lower than those of non-stressed mice in the N-FO group. In contrast with these results in the N-FO group, there was no difference in serotonin levels between SDS-exposed and non-stressed mice in the FO groups. In addition, the expression of genes related to TRP metabolism in SDS-exposed mice increased in the N-FO group, but not in the FO group. These results suggest that fish oil improves the psychosocial behavioral disorders caused by SDS. This improvement could be explained by the increase in serotonin synthesis in the hippocampus.