The attenuation of bluetongue virus by serial passage through fertile eggs

Abstract

1. A strain of virus (University Farm strain) was adapted to propagation in the developing chick embryo by incubation of infected eggs containing 8 day embryos at 33.6°C. but not at 32.1°C., 35.0°C. or 38.2°C. 2. After 3 serial passages at 33.6°C. it was possible to continue propagation at 32.1°C. and 35.0°C. but not at 38.2°C. 3. Using death of the embryos as an index of multiplication of egg-adapted virus there was little difference in the results obtained from incubation at 32.1°C. or 33.6°C. except that multiplication was slightly retarded at the lower temperature. At 35.0°C. the number of survivors beyond the 4th day of incubation was significantly increased. 4. There was little variation in the titre of emulsions produced from dead embryos at either of the temperatures after adaptation to eggs by serial passage. 5. The highest titre emulsions (not less than 10⁻⁵) together with the highest death rate on the 3rd day were produced from eggs incubated for 24 hours at 35.0°C. and then transferred to 32.1°C. 6. The virulent strain of virus was attenuated by serial egg to egg passage. At 32.1°C. attenuation took place rapidly after approximately 20 passages, at 33.6°C. at approximately the same rate, but at 35.0°C. it was delayed until about the 100th subculture. 7. Whether the attenuated virus produces a clinical reaction or not a solid immunity is produced against the homologous strain of virus. 8. The application of the results to the production of large quantities of vaccine for the mass immunization of sheep in the field is discussed.