Objective To study the impaired-welfare induced by routine experimental procedures on mice and the effect of music intervention.Methods 3week male ICR mice were divided into 9 groups. The experimental groups were treated by several experimental operations including restraint(E-r), intragastric injection(E-g), intraperitoneal injection(E-p) and orbital venous sampling(E-o). The music intervention groups were given 1 hour music intervention after the same experimental operation(M-r,M-g,M-p,M-o).After 7 days of continuous treatment, the main neuroendocrine, immune, biochemical parameters, body weight and food efficiency ratio(FER) were measured.Results ①Compared with the untreated group, the body weight gain and FER were significantly decreased in such four experimental groups; TG,HDL-C, WBC,IL-2, β-EP, spleen index and thymus index were significantly decreased in the E-r group, in addition, CORT、ACTH and EPI were significantly increased;TG, WBC, IL-2 and β-EP were significantly decreased in the E-g group, and CORT, ACTH, EPI were significantly increased; IL-2was significantly decreased in the E-p group, and CORT, ACTH, EPI were significantly increased; The spleen index and WBC were significantly decreased in E-o group, and CORT was significantly increased. ②Compared with the experimental groups, the body weight gain, FER, spleen index, IL-2 were significantly increased in the M-r group, and CHOL, CORT, ACTH, β-EP were significantly decreased; WBC was significantly increased in the M-g group, and CORT was significantly decreased; WBC was significantly decreased in the M-p group; FER was significantly increased in the M-o group. Conclusion The four routine experimental procedures cause strong stress response in mice, and lead to health and welfare impaired.However, music intervention played an important role in stress injury improvement.