A survey was performed on ornamental fish imported into the EU to detect viral agents belonging to the genus Ranavirus. The objective was to gain knowledge of the potential for these systemic iridoviruses to gain entry into the EU via international trade in ornamental fish. A total of 208 pooled samples, representing 753 individual fish, were tested. The samples included 13 orders and 37 families, originating from different countries and continents. Tissues from fish that died during or just after transport were collected and examined by standard virological techniques in epithelioma papulosum cyprini cells, by transmission electron microscopy and by PCR for the detection of the major capsid protein and DNA polymerase gene sequences of ranaviruses. Virus was isolated from nine fish species but ranavirus was not identified in those samples. The results suggest that ranaviruses are not highly prevalent in ornamental fish imported into the EU.
- MeSH
- buněčné linie virologie MeSH
- DNA-dependentní DNA-polymerasy analýza genetika MeSH
- Evropská unie MeSH
- fylogeneze MeSH
- infekce DNA virem genetika veterinární MeSH
- karcinom virologie MeSH
- nemoci ryb virologie MeSH
- polymerázová řetězová reakce MeSH
- Ranavirus klasifikace enzymologie genetika ultrastruktura MeSH
- ryby virologie MeSH
- transmisní elektronová mikroskopie MeSH
- virové plášťové proteiny analýza genetika MeSH
- virové proteiny analýza genetika MeSH
- zvířata MeSH
- Check Tag
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
A rabbit polyclonal antibody (PAb) raised against European catfish virus (ECV; isolated from black bullhead Ameiurus melas in France) was produced and then evaluated using a panel of 9 ranavirus isolates collected from different lower vertebrate species originating from Australia, North and South America, Southeast Asia, and Europe. Using ranavirus-infected epithelioma papillosum cyprini (EPC) cell cultures, the specificity of the PAb was determined by Western blot, immunogold electron microscopy, and direct enzyme-linked immunosorbent assay (ELISA). Western blot analysis demonstrated that the PAb reacted strongly with a protein with a molecular weight corresponding to approximately 49 kDa. Immunogold electron microscopy provided direct evidence that the epitopes recognized by this PAb were located on the outer surface of virions. The PAb was used for the preparation of a peroxidase-labeled conjugate for the direct ELISA detection of ranaviruses in infected EPC cell cultures. The specificity of the conjugated PAb was tested using ranaviruses, some representative fish viruses of the genera Rhabdovirus and Birnavirus, and samples from various non-infected fish species. The PAb detected all tested ranaviruses except for 2 Santee-Cooper ranaviruses. The direct ELISA enabled the detection of ranavirus from a concentration of 10(3.5) to 10(3.8) TCID50 ml(-1) cell culture. The results of this study revealed that the rabbit PAb raised against ECV could be useful for the development of specific and standardized diagnostic assays for the detection of ranaviruses from freshwater fish and amphibians.
