The Efficacy of Nile Tilapia Vaccination by Formalin killed Streptococcus agalactiae Encapsulated in Alginate Microcapsules

Abstract

Nile tilapia is one of the major farmed fish that is often infected with Streptococcus. Oral vaccination is the most preferable technique, but the antigen is usually degraded by gastric juice. Encapsulation by alginate could be effective in protecting the antigen. The aim of this study was to assess the effectiveness of vaccinating Nile tilapia using formalin-killed Streptococcus agalactiae encapsulated in alginate. The average diameter of fabricated microcapsules was 500±20 μm, with 86% encapsulation efficacy. Fish were then divided into four groups (30 fish per group): group A: fish vaccinated by alginate microcapsules covered by formalin-killed cells (FKC), group B: fish vaccinated by non-covered FKC, group C: fish fed with empty alginate microcapsules and group D: fish feed with commercial pellets as control group. All groups were treated for 14 days and then fed with commercial pellets. Blood samples were collected on days 7, 14, 21, and 28 days post-vaccination. Bactericidal activity, lysozyme activity and serum antibody titer were significantly elevated in group A (p<0.001) compared to other treated groups. Then, gene expression analysis was performed at 29 days post-vaccination to evaluate the expression of main immune contributors such IgM, IgT, TCR β, CD4, CD8α, IL 1β, IL 8, IFN 1, and TNFα. The analysis showed significant upregulation of all tested genes in group A compared to other treated groups (p<0.001). Lastly, a challenge test was done on day 30 post-vaccination by injecting 4.6×106 CFU mL−1 of virulent S. agalactiae. The relative percent of survival (RPS) were 92± 2%, 48± 5% and 4± 3% for groups A, B and C respectively. The obtained result indicated that alginate encapsulation provided antigen protection due to its higher immunogenicity compared to non-covered vaccine. Hence, the fabricated vaccine could be incorporated with food and orally administered to Nile tilapia to prevent S. agalactiae infection.