Immunization regimen in Asian sea bass (Lates calcarifer) broodfish: A practical strategy to control vertical transmission of nervous necrosis virus during seed production

Abstract Outbreaks of viral nervous necrosis (VNN) in Asian sea bass ( Lates calcarifer ) at the larval stages via vertical transmission of nervous necrosis virus (NNV) from asymptomatic broodfish remain as a major deterrent during seed production. A five-year study was conducted to produce NNV-specific-free sea bass broodfish reared in land-based tanks through an annual immunization regimen with the formalin-inactivated NNV. We primarily immunized (intraperitoneal injection) sea bass juveniles (5 g) and monitored the neutralizing antibody (Nab) titers in the sera of these fish at scheduled intervals post-immunization. Nab titers in the sera of immunized fish peaked at Month 2 (titer: 1:4480 ± 1185) but thereafter gradually declined and significantly dropped (1:260 ± 83) at Month 12 post-primary immunization. Booster immunization of these fish at Month 12 post-immunization led to abrupt increases in Nab titers in booster immunized (B-Im) fish at Month 1 (1:12800 ± 6704) but thereafter declined and dropped at Month 12 (1:480 ± 165) post-booster immunization. The annual booster injections with the inactivated vaccine or L-15 (Unimmunized [U-Im]) were consecutively conducted for 4 years until the fish became sexually mature. Mature fish from both groups were successively induced to spawn twice (1-month interval) via intramuscular injection with luteinizing hormone-releasing hormone analogue (LHRH-a; 100 µg/kg BW). NNV was not detected by RT-PCR in oocytes and milts, and spawned eggs of B-Im fish. In contrast, oocytes and milts, and spawned eggs of U-Im fish were NNV positive. Spawned eggs of B-Im broodfish exhibited Nab titers ranging from 1:192 ± 34 to 1:240 while such was not detected (