Large-scale production and plaque titration of European Chlorella viruses

Viruses of the exsymbiotic green freshwater algae Chlorella, family Phycodnaviridae, appear to be distributed worldwide but those found in North American algae have been characterized in detail. The distinct European Chlorella viruses were studied and it was necessary to adapt both large scale purification and the plaque titration assay to the host organisms' different physiology and to our specific laboratory needs. In the virus purification scheme, a precipitation step with polyethylene glycol was introduced which allows high yield recovery of infective particles from large volumes by rapid low-speed centrifugation. In the plaque assay, a standardized algal culture was introduced. The influence of other factors, e.g. circadian rhythm, on plaque growth is also described.