A protein of 34 kDa (Nx) was induced in vitro by the infection of fish cell cultures with the rhabdovirus agent of viral haemorrhagic septicaemia (VHS) of the trout. This protein only appeared as a major component in concentrated or intracellular labeled VHS virus but not in purified VHS or in the related infectious haematopoietic necrosis virus. That Nx protein is antigenically related to the nucleoprotein of purified virus was shown by its reaction with four anti-nucleoprotein monoclonal antibodies (at least 3 of them reacting non competitively against different epitopes) and by immunoprecipitation with polyvalent international reference sera. The Nx protein was shown to be specifically associated with free non-infective particles isolated by ultracentrifugation which were confirmed to be nucleocapsids by electron microscopy.
At least 28 polypeptides have been identified in intracellular virus, with molecular weights ranging from 11,500 to 243,000 daltons. By treatment with Nonidet P-40 and 2-mercaptoethanol it is possible to obtain subviral particles that have lost some proteins and have a density in CsCl of 1.31 g/cm3 which is higher than that of the complete virus (1.23 g/cm3). After addition of NaCL the virus loses its major protein VP73 which indicates that it is localized in the viral envelope. Cores obtained after this treatment are made up of at least 14 proteins. Incorporation of 3H-fucose and 3H-glucosamine in intracellular virus occurs in three minor components. The protein VP42 is possibly the cell actin and appears to be strongly associated with the virus. It is not possible to eliminate it under conditions where the viral envelopes disappear morphologically. At least the proteins VP172, VP162, VP146 and VP73 act as antigens in the natural infection.
Summary
In this paper the characterization of a canine‐coronavirus (CCV) strain isolated in Spain is reported. The CCV cellular‐infection cycle on A‐72 cells was studied using electron‐microscopy techniques. The isolate was found to have similar electrophoretic profile and antigenic properties to an American reference strain. Sera samples were obtained from several canine populations and tested for viral antibodies using two immunoenzymatic methods. Sera specificity was confirmed using immunoblot analysis. The agreement between both ELISA tests produced adequate results (kappa value = 0.64).
The ultrastructure of natural myxospores and glycerol-induced myxospores has been studied in four Archangium gephyra strains (AG3, AG5, AG9, and AG10). The induced myxospores show an oval shape, characteristic of the genus Archangium, and an electron-dense coat surrounds the cell wall. The cytoplasm contains abundant electron-lucent inclusions. Sometimes, a membranous structure appears between the coat and the outer membrane. The fruiting-body myxospores show an oval shape similar to that observed in induced myxospores, but with a slightly thicker outer coat. In many of the natural myxospores, some membranous, lamellar or vesicular structures appear. They are located between the coat and the outer membrane, resembling those that appeared in induced myxospores. These have not been previously mentioned in papers on fruiting-body myxospores. These structures occasionally seem to be fused, giving rise to membranous sacs filled with an amorphous material of medium electron density. We propose that these vesicles are formed from the waved outer membrane and from the fruiting-body matrix. Large, dense, fusiform inclusions with an internal crystalline array are frequently found in the cytoplasm of natural myxospores, and cells during differentiation, in strains AG3, AG9, and AG10. Key words: Archangium gephyra, electron microscopy, myxospores, ultrastructure.
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