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Title: QUANTITATIVE REAL-TIME REVERSE TRANSCRIPTION-PCR ANALYSIS OF DEFORMED WING VIRUS INFECTION IN THE HONEYBEE (APIS MELLIFERA L.)

Author
item Chen, Yanping - Judy
item Higgins, James
item Feldlaufer, Mark

Submitted to: Applied and Environmental Microbiology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/4/2004
Publication Date: 8/4/2004
Citation: Chen, Y.P., Higgins, J.A., Feldlaufer, M.F. 2004. Quantitative real-time reverse transcription-pcr analysis of deformed wing virus infection in the honeybee (apis mellifera l.). Applied and Environmental Microbiology.71(1):436-441.

Interpretive Summary: Deformed wing virus (DWV) is a honey bee virus that has recently been found in the United States. When we tested honey bees for DWV, both bees with deformed wings and those with apparently normal wings contained virus. Using molecular techniques we determined the amounts of virus in individual bees representing these two groups. Not surprisingly, honey bees with deformed wings contained higher levels of virus than bees with normal wings. We also determined virus levels in immature bees to determine if levels in immature bees could be used to predict whether or not adult bees would have symptoms. The methodologies described in this research will be used as a diagnostic tool by other scientists to determine levels of virus necessary to adversely affect bees.

Technical Abstract: Deformed wing virus (DWV) is a honey bee virus that can cause wing deformity and premature death in adults, though like many other bee viruses, DWV generally persists as a latent infection with no apparent symptoms. Using RT-PCR and Southern hybridization, we detected DWV in all life stages of honey bees including adults, with and without deformed wings. We also found DWV in the parasitic mite, Varroa destructor, suggesting this mite may be involved in the transmission of DWV. However, the detection of virus in life stages not normally associated with mite parasitism, i.e. eggs and larvae, suggests other modes of transmission. Levels of DWV in different life stages of bees were investigated by TaqMan real time quantitative RT-PCR. Amounts of virus varied significantly in these different stages, with the highest levels occurring in pupae and adult worker bees with deformed wings. Variability in virus titer may reflect the different abilities of bees to resist DWV infection and replication. The epidemiology of DWV is discussed and factors such as mite infestation, malnutrition, and climate are considered.