In this study, used grouper GF-1 cells to proliferate grouper nervous necrosis virus and prepare inactivated vaccine . These NNV inactivated vaccine had been immunized the SPF hens., after multiple boost, the ELISA analysis of the titers of anti-NNV IgY could up to 104 ~ 105 dilutions. The crude egg yolk and freeze-dried egg yolk powder were conducted neutraliation test , the antibody titers get up to 219 and 216 dilutions, respectively. In the challenge model, the larvae of grouper were pre-fed the feed containing anti-NNV IgY 4 days before challenged 105 PFU/ml NNV. The survival rate of the larvae of grouper was 60%. These data showed that the passive immunity feed containing anti-NNV IgY could provide the protective immunity of the larvae of grouper against NNV infection.