In recent years, there has been a growing focus on using herbal extracts as immune enhancers for aquatic species, replacing antibiotics. In the present study, the effects of dietary supplementation of Hericium erinaceus extract (HE) on growth, feed utilization, hematology, expression of immunity-related genes, and immune responses in Nile tilapia infected by Streptococcus agalactiae were examined. A total of 240 Nile tilapia with an average body weight of 17.28 ± 0.01 g were fed diets enriched with different levels of HE: 0 (HE0), 0.1 (HE0.1), 1.0 (HE1.0), and 5.0 (HE5.0) g/kg. The results showed that growth parameters, feed conversion ratio, and organosomatic indexes were not linearly or quadratically affected by HE supplementation. Fish fed HE0.1 and HE1.0 increased protein efficiency ratio and protein productive values with significant linear and quadratic effects of HE enrichment. In addition, dietary supplementation of HE quadratically increased whole-body protein content. Red blood cell, white blood cell, and hematocrit were linearly and quadratically increased by HE supplementation. HE also linearly and quadratically decreased LDL cholesterol and linearly decreased the total cholesterol levels. Stress markers, serum glucose, and cortisol levels were linearly and/or quadratically decreased in HE-fed fish. The relative mRNA expression of tnf-α, il-1β, il-6, and il-10 were upregulated in the HE0.1 and HE1.0 groups, while dietary supplementation of HE significantly decreased hsp70cb1 mRNA expression in all groups. After feeding dietary HE supplementation for 10 weeks, fish were intraperitoneally injected with pathogenic S. agalactiae. A high survival after challenge was found in all HE supplementation groups with the highest percent survival observed in the HE1.0 and HE5.0 groups. Our findings represent that supplementation of 1 g/kg of HE (HE1.0) could obtain the greatest effects on immunity and survival of Nile tilapia. In addition, the present study also showed that dietary supplementation of HE can improve protein utilization, hematology, expression of genes related to immunity, stress markers, and resistance of Nile tilapia against pathogenic bacterial infection.