Protein pI alteration related to strain variation of infectious bronchitis virus, an avian Coronavirus.

Viral proteins of two strains of infectious bronchitis virus (IBV), which have different tissue trophism and serology, were separated on the basis of their isoelectric points (pI). The viruses have four structural proteins; the protein of greatest serological importance is found at the peplomer tip. The viral structural proteins separated by isoelectric focusing were identified by comparison to SDS-PAGE separations. Three protein bands were identical in pI and one protein band showed a difference in pI between strains. When the renatured viral proteins were Western blotted and reacted with strain-specific antiserum, antigen-antibody complexing was seen only at points corresponding to the strain-specific variant bands. For IBV strain Mass-41, antigen-antibody complexing occurred at a pI of 6.8, and, for IBV strain Ark-99, at 7.2. No cross reaction of antisera was observed for either strain. Since tissue affinities are a function of the viral peplomer-mediated attachment of virus to cells and are often directly related to pathogenicity, it appears that altered pathogenicity of strains of IBV may be detected by alteration of pI of the proteins. Classification by pI of proteins of at least the smaller viruses allows untypeable, highly pathogenic or persistent strains of these viruses to be characterized on the basis of variant proteins.

Detection of antibody to avian viruses in human populations.

The ability of three avian viruses to elicit antibody response in humans was surveyed for the purpose of identifying zoonotic diseases. Antibody levels in people associated with poultry were compared to those in people having limited poultry association. Antibody levels to three avian viruses: infectious bursal disease virus, a birnavirus; Newcastle disease virus, a paramyxovirus; and avian infectious bronchitis virus, a coronavirus were determined by enzyme-linked immunosorbent assays (ELISA). Differences between the two study groups were evident: people having a known association with poultry showed significantly higher levels of antibodies to Newcastle disease and avian infectious bronchitis virus. Antibodies detected may be due to virus exposure rather than zoonoses.

Detection of latent avian adenovirus-associated virus proteins in chicken cells.

Chicken kidney cells, derived from the eggs of white leghorn chickens that had serological evidence of prior exposure to both adenovirus and the adeno-associated virus (AAV), produced AAV antigenic proteins upon challenge with purified adenovirus. Antigen was detected by indirect immunofluorescence using monoclonal antibody to AAV. The number of fluorescent cells were few and did not increase during the course of adenovirus infection. Similar results were obtained using cells prepared from specific-pathogen-free chicks with no previous exposure to adenovirus or AAV. It is postulated that the avian AAV can exist as a latent infection in the germ line of chickens.

Morphogenesis of canary poxvirus and its entrance into inclusion bodies.

A virus isolated from a natural outbreak of canarypox was replicated on the chorioallantoic membranes of chicken embryos, and its ultrastructure and development were observed. Electron microscopy of thin sections of pocks produced on the chorioallantoic membranes revealed a variety of developmental forms which appear similar to those demonstrated in studies of vaccinia, ie, viroplasm or viral factories; immature, undifferentiated virions partially enclosed by membranes; completely enclosed nondifferentiated spherical or oval virions; immature virions with discrete nucleoids; and the more compact brick-shaped mature virions. Two types of A-type inclusions were noted: those with virions around the periphery, and those filled with virus particles. The appearance of mature viruses within the inclusion bodies and different stages of viruses outside the inclusion indicate that in a course of development, maturing poxvirus may enter the inclusion bodies as they acquire surface tubules on their envelopes. Mature virions also were seen budding out of the cell membrane, apparently enveloped in a portion of the membrane. Studies showing the entrance of poxvirus into inclusion bodies have not been reported. In this report, electron micrographs are shown of viruses entering inclusion bodies.

The effect of incubation temperature on the propagation of duck adenovirus (virus 127-like) in duck and chicken cells.

Duck adenovirus (Cornell strain) was propagated in duck and chicken embryo cells at 37.5 C and at 40 C. In duck cells, virus levels, as indicated by HA titers, peaked earlier at 40 C than at 37.5 C. High titers were eventually observed in duck cells at both temperatures. In chicken embryo fibroblasts, no titers were observed at 37.5 C, whereas low titers were observed at 40 C. Evidence of virus propagation was not detected in chicken embryo liver and kidney cells.