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RT-PCR analysis of Deformed wing virus in honeybees (Apis mellifera) and mites (Varroa destructor)
Constanze Yue1, Elke Genersch1
1Institute for Bee Research, Friedrich-Engels-Str. 32, D-16540 Hohen Neuendorf, Germany.
Abstract:
Deformed wing virus (DWV) is a honeybee viral pathogen either persisting as an inapparent infection or resulting in wing deformity. The occurrence of deformity is associated with the transmission of DWV through Varroa destructor during pupal stages. Such infections with DWV add to the pathology of V. destructor and play a major role in colony collapse in the course of varroosis. Using a recently developed RT-PCR protocol for the detection of DWV, individual bees and mites originating from hives differing in Varroa infestation levels and the occurrence of crippled bees were analysed. It was found that 100 % of both crippled and asymptomatic bees were positive for DWV. However, a significant difference in the spatial distribution of DWV between asymptomatic and crippled bees could be demonstrated: when analysing head, thorax and abdomen of crippled bees, all body parts were always strongly positive for viral sequences. In contrast, for asymptomatic bees viral sequences could be detected in RNA extracted from the thorax and/or abdomen but never in RNA extracted from the head. DWV replication was demonstrated in almost all DWV-positive body parts of infected bees. Analysing individual mites for the presence of DWV revealed that the percentage of DWV-positive mites differed between mite populations. In addition, it was demonstrated that DWV was able to replicate in some but not all mites. Interestingly, virus replication in mites was correlated with wing deformity. DWV was also detected in the larval food, implicating that in addition to transmission by V. destructor DWV is also transmitted by feeding.
Insights
Deformed wing virus (DWV) infects all honeybees, but crippled bees show higher viral loads in all body parts. Virus replication in Varroa mites correlates with wing deformity, suggesting a key role in honeybee colony collapse.
Area of Science:
- Apiculture
- Virology
- Entomology
Background:
- Deformed wing virus (DWV) is a significant honeybee pathogen.
- DWV infection can be inapparent or cause wing deformities, particularly when transmitted by Varroa destructor mites during pupal stages.
- DWV contributes to Varroa mite pathology and honeybee colony losses.
Purpose of the Study:
- To investigate the spatial distribution and replication of DWV in honeybees and Varroa mites.
- To determine the correlation between DWV presence in mites and honeybee wing deformity.
- To explore alternative DWV transmission routes beyond Varroa mites.
Main Methods:
- Utilized a novel RT-PCR protocol for DWV detection in individual bees and mites.
- Analyzed DWV presence and replication in different body parts (head, thorax, abdomen) of crippled and asymptomatic bees.
- Quantified DWV-positive mites from hives with varying infestation levels and deformity incidence.
- Detected DWV in larval food samples.
Main Results:
- All tested bees, both crippled and asymptomatic, were DWV-positive.
- Crippled bees showed strong DWV presence in all body parts, while asymptomatic bees had it only in the thorax and/or abdomen, never the head.
- DWV replicated in most infected bee body parts and in some Varroa mites.
- DWV presence and replication in mites correlated with honeybee wing deformity.
- DWV was also found in larval food, indicating transmission through feeding.
Conclusions:
- DWV distribution and replication patterns differ between symptomatic and asymptomatic honeybees.
- Varroa mite infestation and DWV replication within mites are strongly linked to honeybee wing deformity.
- Larval food represents an additional route for DWV transmission in honeybee colonies.
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