Protective immunity against viral nervous necrosis (VNN) in brown-marbled grouper (Epinephelus fuscogutattus) following vaccination with inactivated betanodavirus
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Viral nervous necrosis (VNN) caused by betanodaviruses has been recently implicated in serious mortalities of groupers in the grow-out culture system. A safe and effective vaccine against this disease is urgently needed. This study demonstrates that a single intramuscular vaccination with formalin-inactivated Philippine strain of piscine betanodavirus (genotype: redspotted grouper nervous necrosis virus; RGNNV) induces potent immune responses and substantial protective immunity against an intramuscular challenge with the homologous virus in brown-marbled grouper, Epinephelus fuscogutattus, a highly susceptible marine fish species to VNN. Seroneutralization assay conducted on sera of vaccinated fish revealed the occurrence of substantial neutralizing-antibody titers from Days 15 (mean titer 1:800) to 190 (1:400) with the highest titer observed at Day 60 post-vaccination (1:5120). When vaccinated fish were challenged with the homologous virus at Days 15, 30 and 75 post-vaccination, significantly higher survival rates were obtained in these fish compared with their corresponding controls (L-15 injected fish). Abrogation of virus multiplication in all vaccinated survivors was indicated by undetectable virus titers in the brains and kidneys paralleled by significantly high levels of neutralizing antibodies in the sera of these fish. Consecutively, replicates of vaccinated fish that survived betanodavirus challenge at Days 15 and 75 post-vaccination were maintained in flow-through aquaria and rechallenged with the homologous virus 3 and 5 months later, respectively. A significant drop in neutralizing-antibody titers of 3 and 8 folds, respectively, were observed in the sera of Days 15 and 75 post-vaccinated fish assayed before the virus rechallenge. Interestingly, reversion in the levels of neutralizing antibodies to significantly high levels (8–15 folds) were noted in these fish after the virus rechallenge. Taken together, our current data clearly demonstrate that a single administration of the inactivated Philippine strain of betanodavirus vaccine can effectively mount a specific anamnestic response and concomitant long-term protection against VNN in grouper at the grow-out culture system.
CitationPakingking Jr., R., Bautista, N. B., de Jesus-Ayson, E. G., & Reyes, O. (2010). Protective immunity against viral nervous necrosis (VNN) in brown-marbled grouper (Epinephelus fuscogutattus) following vaccination with inactivated betanodavirus.
This study was funded by SEAFDEC AQD (study code: 5203-T-RDFH0507) and in part by the Government of Japan Trust Fund through the Regional Fish Disease Project (study code: 8001-T-FD-FH0507). We would like to thank Dr. Felix G. Ayson, Chief Technical Adviser, Food and Agriculture Organization of the UN (FAO), Fish Farming Center, Jeddah, Saudi Arabia for the critical reviewof the manuscript.
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Susceptibility of hatchery-reared snubnose pompano Trachinotus blochii to natural betanodavirus infection and their immune responses to the inactivated causative virus Mass mortality of snubnose pompano Trachinotus blochii fry exhibiting dark coloration, anorexia, and abnormal swimming behavior was recently documented at the hatchery of the Aquaculture Department of the Southeast Asian Fisheries Development Center, Philippines. Samples of brain tissues were collected from affected fish and processed for RT-PCR amplification and virus isolation in cell culture. Infected E-11 cells exhibited cytopathic effect characteristic of betanodavirus. Histopathology of moribund fish showed pronounced vacuolations in the brain, spinal cord, and retina. An RT-PCR product of approximately 430 bp was amplified from the culture supernatant of betanodavirus-infected E-11 cells and sequenced. Sequencing of the T4 region of the coat protein gene (RNA 2) revealed clustering of the isolated virus within the red-spotted grouper nervous necrosis virus type. The pathogenicity of the isolated betanodavirus in healthy pompano juveniles and fry was determined via intramuscular injection and immersion challenges, respectively. Higher mortality rates were obtained in challenged fish compared with the controls. An inactivated vaccine was subsequently prepared by treating the clarified betanodavirus with formalin. Pompano juveniles intraperitoneally injected with the inactivated vaccine exhibited neutralizing antibodies from days 15 (mean titer 1:240) to 125 (1:560) with the highest titer noted at day 64 (1:2240) post-vaccination. Additionally, pompano fry bath-vaccinated and consequently bath-challenged with betanodavirus at day 35 post-vaccination showed higher survival rate compared with the control, indicating the potential of the inactivated betanodavirus vaccine against VNN in pompano fry and juveniles.
Immune responses of Asian sea bass, Lates calcarifer Bloch, against an inactivated betanodavirus vaccine RV Pakingking Jr., R Seron, LD de la Peña, K Mori, H Yamashita & T Nakai -
Journal of Fish Diseases, 2009 - Blackwell PublishingAsian sea bass, Lates calcarifer (Bloch), exhibited strong immune responses against a single injection of the formalin-inactivated red-spotted grouper nervous necrosis virus (RGNNV), a betanodavirus originally isolated in Japan. Fish produced neutralizing antibodies at high titre levels from days 10 (mean titre 1:480) to 116 (1:1280), with the highest titre at day 60 post-vaccination (1:4480). When fish were challenged with the homologous RGNNV at day 54 post-vaccination, there were no mortalities in both the vaccinated and unvaccinated control fish. However, a rapid clearance of the virus was observed in the brains and kidneys of vaccinated fish, followed by a significant increase in neutralizing-antibody titres. Furthermore, the vaccine-induced antibodies potently neutralized Philippine betanodavirus isolates (RGNNV) in a cross-neutralization assay. The present results indicate the potential of the formalin-inactivated RGNNV vaccine against viral nervous necrosis (VNN) of Asian seabass.
Immuno-response in tilapia Sarotherodon niloticus vaccinated with Edwardsiella tarda by hyperosmotic infiltration method Sarotherodon niloticus with average weight of 28.42 ± 1.87g were immunized with formalin-killed Edwardsiella tarda using the hyperosmotic infiltration method. Test fish maintained in 30 l aquaria were grouped into four treatments. Group 1 and 2 were exposed to a single hyperosmotic treatment on day 0. Group 1 was bled on day 14 and group 2 was bled on day 28. Group 3 was given hyperosmotic treatments twice: on day 0 and day 14 and bled on day 28. Group 4 was an untreated control bled on day 28. All sera were analyzed for agglutinating antibody titer against E. tarda flagellar and somatic antigens. Results showed that flagellar and somatic agglutinin titers in all treatments were not statistically significant. Likewise, infection experiments where test fish were challenged with intraperitoneal injection of the test bacterium showed that the vaccination experiment did not effectively protect the test fish from infection by Edwardsiella tarda.