Location: Bee Research LaboratoryTitle: Beeporter: a high-throughput tool for non-invasive analysis of honey bee virus infection
|BANMEKE, OLUBUKOLA - Orise Fellow|
|PALMER-YOUNG, EVAN - Orise Fellow|
|Chen, Yanping - Judy|
|RIABOV, EUGENE - Volunteer|
Submitted to: Molecular Ecology Resources
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/22/2021
Publication Date: 1/5/2022
Citation: Evans, J.D., Banmeke, O., Palmer-Young, E., Chen, Y., Riabov, E. 2022. Beeporter: a high-throughput tool for non-invasive analysis of honey bee virus infection. Molecular Ecology Resources. 22:978–987. https://doi.org/10.1111/1755-0998.13526.
Interpretive Summary: Honey bee losses are unsustainably high. Disease agents, from mites to viruses and gut parasites are responsible for a large fraction of these losses So far there are no registered treatments to control bee viruses. The described protocol will speed up the search for such treatments, allowing researchers to convert treatment candidates into new bee drugs more quickly. Since virus levels can be monitored without killing bees, this method can also be used to screen bees for viral resistance as part of breeding schemes. More generally, this method will work for other insects species to study virus movement and growth in pest and beneficial insects.
Technical Abstract: Honey bees face numerous pests and pathogens but arguably none are as devastating as Deformed wing virus (DWV). Development of antiviral therapeutics and virus-resistant honey bee lines to control DWV in honey bees is slowed by the lack of a cost-effective high-throughput screening of DWV infection. Currently, analysis of virus infection and screening for antiviral treatments in bees and their colonies is tedious, requiring a well-equipped molecular biology laboratory and the use of hazardous chemicals. Here we utilize a cDNA clone of DWV tagged with green fluorescent protein (GFP) to develop the Beeporter assay, a method for detection and quantification of DWV infection in live honey bees. The assay involves infection of honey bee pupae by injecting a standardized DWV-GFP inoculum, followed by incubation for up to 44 hours. GFP fluorescence is recorded at intervals via commonly available long-wave UV light sources and a smartphone camera or a standard ultraviolet transilluminator gel imaging system. Nonlethal DWV monitoring allows high-throughput screening of antiviral candidates and a direct breeding tool for identifying honey bee parents with increased antivirus resistance. For even more rapid drug screening, we also describe a method for screening bees using 96-well trays and a spectrophotometer.