Systemic Spread and Propagation of a Plant-Pathogenic Virus in European Honeybees, Apis mellifera

Li, Ji Lian; Cornman, R. Scott; Evans, Jay D.; Pettis, Jeffery S.; Zhao, Yan; Murphy, Charles; Peng, Wen Jun; Wu, Jie; Hamilton, Michele; Boncristiani, Humberto F.; Zhou, Liang; Hammond, John; Chen, Yan Ping

Honeybee colonies and sample collection. Honeybee colonies used for this study were maintained in the research apiaries of the USDA-ARS Bee Research Laboratory in Beltsville, MD. For viral particle purification and tissue dissection, fifty adult worker bees were collected by removing a central frame filled with brood and covered with adult bees from a hive and gently scraping worker bees into a 50-ml conical tube. In addition, bee pollen that was processed by bees and stored in combs around the brood was collected using a spatula and transferred into 15-ml conical tubes. Individual Varroa mites that had crawled from brood cells onto the tops of brood frames were collected with forceps and transferred into 1.5-ml microcentrifuge tubes.