Browsing by Subject "Immunofluorescence"
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Bulletin of the European Association of Fish Pathologists, 2008 - European Association of Fish PathologistsViral nervous necrosis caused by betanodaviruses is one of the most devastating diseases in cultured marine finfish. In the Philippines, mass mortalities occurred in sea bass, Lates calcarifer larvae and grouper, Epinephelus coioides broodstock. The virus was isolated using SSN-1 fish cell line and confirmed by PCR. Cytopathic effect started to develop in the cell line 2 days post infection (p.i) with tissue filtrates until the cells completely disintegrated and detached from the flask at 5 days p.i. and the viral protein was detected by immunofluorescence. Sequence analysis revealed that VNN isolated from the brain of grouper broodstock and sea bass larvae were 98.6% similar. Sequence analysis between the Philippine isolates and red-spotted grouper nervous necrosis virus (RGNNV) genotype is 96.9% similar as compared to 72.0% and 64.0% similar with the barfin flounder nervous necrosis virus (BFNNV) and tiger puffer nervous necrosis virus (TPNNV) genotypes, respectively. These results confirm that the Philippine isolates belong to RGNNV genotype.
Conference paper- In Y Inui & ER Cruz-Lacierda (Eds.), Disease Control in Fish and Shrimp Aquaculture in Southeast Asia - Diagnosis and Husbandry Techniques: Proceedings … Aquaculture in Southeast Asia - Diagnosis and Husbandry Techniques, 4-6 December 2001, Iloilo City, Philippines, 2002 - Aquaculture Department, Southeast Asian Fisheries Development CenterThe first outbreak of red sea bream iridoviral disease (RSIVD) caused by red sea bream iridovirus (RSIV) was recorded among cultured red sea bream (Pagrus major) in 1990 in Ehime, Shikoku, Japan. Since then, the disease has caused mass mortalities of many cultured marine fishes. From 1990-2000, RSIVD was detected in 31 cultured marine fish species, including 28 Perciformes, 2 Pleuronectiformes and 1 Teteraodontiformes, in 18 prefectures in the southwestern part of Japan. The infected fish are lethargic and show severe anemia, petechiae of the gills, and enlargement of the spleen. Histopathologically, the disease is characterized by the presence of enlarged cells in the spleen, heart, kidney, liver and gills that are deeply stained with Giemsa solution. Diagnostic methods for RSIV, such as the observation of stained imprints or tissue sections, an immunofluorescent (IF) test with a monoclonal antibody (MAb) and a polymerase chain reaction (PCR) technique have been developed. The IF test with MAb is commonly used in the rapid diagnosis of RSIV-infected fish. For an effective control measure against RSIVD, a formalin-killed vaccine has been developed and this showed a significant effect in red sea bream under both experimental and field conditions.
Immunolocalisation of nervous necrosis virus indicates vertical transmission in hatchery produced Asian sea bass (Lates calcarifer Bloch)—A case study -
Aquaculture, 2006 - ElsevierA probable vertical mode of piscine nodavirus transmission is reported in the present investigation based on a case of nodavirus associated larval mortalities in hatchery produced Asian sea bass. Polyclonal rabbit anti-SJNNV antibodies (SGWak97) detected the viral antigens in the tissue sections from the eggs and the larvae at different time intervals from − 1 to 42 days post hatch (dph). Immunopositive ovarian connective tissue associated with the oocytes along with the progressive localization of the viral antigens in the brain, spinal cord, liver, stomach and dermal musculature during the larval development indicates a probable vertical transmission of nodavirus in the Asian sea bass. The surviving larger larvae, from the batch suffering mass mortalities, produced very intense immunofluorescent positivity in the liver, stomach and dermal musculature. Results of this investigation demonstrating a possibility of vertical transmission of the nodavirus emphasize the need for screening of eggs and larvae for evolving suitable preventive and prophylactic health management strategies.