Hydrogen peroxidase-inactivated vaccines have recently developed in controlling infectious diseases in aquaculture. The present study aimed to compare the efficacy of the hydrogen peroxidase-inactivated Streptococcus agalactiae vaccine via different delivery routes (injection, immersion, and oral administration) in red tilapia (Oreochromis sp.) without using adjuvant. Fish were randomly divided into 4 groups including G1: Control treatment (without vaccine), G2: vaccine-based diet (oral administration), G3: vaccinated by immersion and G4: vaccinated by injection. After 6 weeks of the experiment, fish were intraperitoneally injected with S. agalactiae and the mortality was recorded in 14 days. The results showed that lysozyme activity was differentially increased according to the delivery routes of vaccine, organs and time of sampling. However, the specific antibody levels in all vaccinated groups were only increased in week 6 post-vaccination. After the challenge test with S. agalactiae, the serum lysozyme levels in G3 and G4 were significantly higher than the control group (G1), while the total white blood cells and specific antibody levels were significantly increased in G2 and G4 compared to the control (G1). Similarly, the hydrogen peroxidase-inactivated vaccine statistically reduced the cumulative mortality in G2 (35.29%) and G4 (28.95%) compared to G1 (44.12%) after injected with the S. agalactiae. These results showed that the vaccine delivery routes by oral administration or injection may decrease the pathogen and show better protection for red tilapia than the immersion method. Further studies will be investigated to improve the efficacy of hydrogen peroxidase- inactivated vaccine via using different adjuvants.