Lumpfish (Cyclopterus lumpus) has become the predominant cleaner fish species used in North American salmon aquaculture for sea lice (e.g., Lepeophtheirus salmonis) biocontrol. Lumpfish utilization has contributed significantly towards eliminating the utilization of chemotherapeutants by effectively controlling the abundance of this damaging pest of Atlantic salmon (Salmo salar) aquaculture. Vibrio anguillarum is a frequent pathogen of lumpfish in Atlantic Canada. Here, several vaccine trials against V. anguillarum were conducted. Fish health, including effective vaccine design, and vaccination programs have been identified as a high priority. In this study, the safety and efficiency of a V. anguillarum autogenous vaccine were evaluated. Five treatments with three replicates were used in this study. They were PBS (negative control), in-house vaccine (positive control), autogenous IP (intraperitoneally), autogenous deep and autogenous deep, and IP boost. Length and weight were taken, gross pathology was conducted. These treatments were bath challenged with V. anguillarum J360 (serotype O2) after 10 weeks post-vaccination, and the survival was bath re-challenged after 9 weeks of the first challenge with V. anguillarum J360 (serotype O2) and V. anguillarum J382 (serotype O1). Tissue samples were collected for tissue colonization count. No mortality after vaccination signified that the vaccine is safe. The gross pathology score indicated a normal immune response. The in-house IP, autogenous IP, and autogenous deep and IP boost had a relative percentage survival (RPS) of 76.12%, 72.12%, and 68.75% respectively in the first challenge. The vaccine only protects against V. anguillarum J360 (serotype O2) but does not confer protection against V. anguillarum J382 (serotype O1).