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A laboratory test system specific for Marek's disease was developed by using the pathological response of the chicken embryo. Chicken epidermal scales (dander) and feather calami from infected chickens contain an agent(s) which after a 3- to 4-day incubation period caused gross or microscopic pathological changes (or both) in the embryo. A cell-free inoculum was obtained from infectious dander by 5-min sonic treatment, differential centrifugation, and membrane filtering (0.45 mum). Evidence for the cell-free existence of this agent(s) was obtained when membrane filtrates of dander preparations were shown to cause Marek's disease in 10-day-old chickens and in chickens inoculated at 1 day of age.

Marek's disease virus recovered from the feather follicle of infected chickens was found to be infectious for chickens in cell-free preparations. The virus replicated in epithelial cells of the germinative layer of the feather follicle epidermis, producing both intranuclear and round or diffuse cytoplasmic inclusion bodies in the infected cells. It was found at this site 2 weeks postinoculation and prior to the development of tumor or other gross lesions. In the nucleus, many naked and a few enveloped herpesvirions were found, whereas the cytoplasm contained predominantly enveloped herpesvirions, which were usually within the cytoplasmic inclusion bodies. Approximately 80% of the extracellular virions were enveloped. Studies with both virulent and avirulent strains of the virus revealed a relationship between virulence, contagiousness, and replication of the virus in the feather follicle.