| Summary: | 碩士 === 國立中山大學 === 海洋生物科技暨資源學系研究所 === 95 === The grouper is a high-value fish in Taiwan. However, nervous
necrosis virus, which contains two single strand RNAs and doesn’t have an
envelope, causes groupers die in early stages of development. It
economically impacts on the aquaculture of this marine fish. Vaccination is
one of the best methods to prevent viral diseases. Virus-like particles
(VLPs) were used for studying the ability and efficiency of producing
antibodies against dragon grouper nervous necrosis virus. Different
injection dosages and injection frequency of VLPs were performed on
Epinephelus lanceolatus. The anti-sera of vaccinated fish were acquired by
microvoluminal phlebotomization method and analyzed by antigen-capture
dot blotting with ECL detections, which are the best choices for qualitative
and quantitative assays. The signal of antibodies in the vaccinated fish was
detected every week after primary immunization. The antibody signal
reached 8.4x106 U in one month when the dragon groupers were injected
with 10 µg of VLPs, but giving additional injection of VLPs didn’t
increase production of antibodies after one month. Moreover, dragon
groupers that were injected with 50 µg and 100 µg of VLPs generated
antibody signals up to 7.2x106 U and 6.7x106 U, respectively. The antibody
signal can remain at a higher level in the presence of high dosage injection
for at least three weeks. Furthermore, a polyvalent vaccine that included
killed bacteria was applied in field to test dragon grouper immune response.
The results not only support the data that dragon grouper boots antibody
production within a short period after immunization, but also demostrates
that the production of antibody by dragon grouper against virus and
bacteria can be monitored by the microvoluminal phlebotomization
method.
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