Project description:This dataset represents RNA-Seq data that was later found to have widespread contamination with Deformed Wing virus (DWV). It has been incorporated in a meta-analysis of DWV's impact on brain gene expression
Project description:This dataset represents RNA-Seq data that was later found to have widespread contamination with Deformed Wing virus (DWV). It has been incorporated in a meta-analysis of DWV's impact on brain gene expression
Project description:Bees from 3 unrelated colonies were injected with 1ul PBS extract containing 10^9 genome equivalent of Deformed wIng virus (DWV) and/or fed 10µl sucrose solution containing 10^5 fresh Nosema ceranae spores. Control bees were injected and fed with an equivalent DWV- and Nosema-free extract respectively). Bees were kept in cages of 21 bees (7 from each colony), and each cage was replicated 5 times per each of the 4 treatments). Bess were kept in an incubator at 30°C/50%RH. At day 12 p.i., bees were flash frozen in liquid nitrogen, and stored at -80°C. Bee abdomen RNA was sent to Christina Grozinger lab (Penn State, USA). RNA was pooled for 3 abdomens per replicate for 5 replicates per treatment. Arrays were hybridized in a dye-swap loop design.
Project description:Experiment was designed (i) to analyse the strain composition of Deformed wing virus (DWV) populations in covertly and overtly infected honeybees (Apis mellifera) from Varroa-free and Varroa-infested colonies, and (ii) to determine abundance of the DWV strains following direct injection of the DWV preparations from covertly and overtly infected bees to the bee pupae haemolymph in the absence of Varroa destructor mites. Experiment included isolation of DWV preparations from the following bees: covertly-infected bees from Varroa-free colony, covertly infected bees exposed orally to the Varroa-selected DWV strains, and the overtly infected Varroa-exposed bees. Honeybee pupae were experimentally injected with those DWV preparations and sampled 4 days post injection following development of overt DWV infection. A series of the DWV cDNA fragment covering complete DWV genomic RNA sequences were amplified by RT-PCR using RNA extracted from virus preparations and the injected pupae. The cDNA preparations were sequenced using next generation(Illumina HighSeq 2000) paired-end sequencing to obtain data on the DWV strain composition.
Project description:Plant pollination by the western honey bee Apis mellifera is an irreplaceable agroecological and economic cornerstone currently under threat. Recent colony loss has been consistently linked to the increased prevalence of deformed wing virus (DWV), an Iflavirus transmitted from the ecoparasitic mite Varros destructor. While DWV has been detected in the honey bee brain and causally linked to behavioral impairment, the molecular impact of infection on brain gene expression is largely unknown. Recently, we discovered that two published and two new brain transcriptomic studies conducted in our lab contained DWV contamination in over 99% of sequenced honey bee samples. This unanticipated finding sharply contrasted with the experimental paradigms of these four studies, as no physical or behavioral signs of DWV were detected in any of the 335 individual honey bees sampled. We took this opportunity to perform a meta-analysis and test the hypothesis that DWV influences brain gene expression, a relationship which could be linked to the massive depopulation events observed around the world. Results from our study support commonalities in the molecular consequences of DWV in the honey bee brain and implicate specific genes and biological processes associated with infection. Next, we used single-cell RNA-Sequencing to implicate glia as active responders to viral infection. Finally, we performed viral gene expression analysis on a subset of samples and found DWV type A as well as a previously unreported A-B recombinant in the brain. We present this meta-analysis as a first step toward addressing a potential missing link between viral infection and behavior in honey bees.
Project description:Experiment was designed to study the effect of Deformed wing virus (DWV) and the mite Varroa destructor on global gene expression using microarray transcriptional profiling in developing worker honeybee (Apis mellifera). Newly hatched bee larvae (day 3 of bee development) were transferred from a Varroa-free colony with low DWV levels to a Varroa-infested colony with high levels of DWV in bees and Varroa mites. All transferred larvae were receiving the DWV strains present in this Varroa-infested colony with the food delivered by the nurse bees until their capping (day 8). About half of these larvae were capped with Varroa mite and were subjected to the mite piercing and feeding on their haemolymph during pupal development until sampling at purple eye stage (day 14). Exposure to the mite piercing and feeding resulted in about 1000-fold increase of the DWV levels in the majority of the mite-exposed pupae compared to the control pupae and the pupae not exposed to Varroa mites.
Project description:Experiment was designed to study the effect of Deformed wing virus (DWV) and the mite Varroa destructor on on siRNA and miRNA composition using high-throughput sequencing of small RNA in developing worker honeybee (Apis mellifera). Newly hatched bee larvae (day 3 of bee development) were transferred from a Varroa-free colony with low DWV levels to a Varroa-infested colony with high levels of DWV in bees and Varroa mites. All transferred larvae were receiving the DWV strains present in this Varroa-infested colony with the food delivered by the nurse bees until their capping (day 8). About half of these larvae were capped with Varroa mite and were subjected to the mite piercing and feeding on their haemolymph during pupal development until sampling at purple eye stage (day 14). Exposure to the mite piercing and feeding resulted in about 1000-fold increase of the DWV levels in the majority of the mite-exposed pupae compared to the control pupae and the pupae not exposed to Varroa mites.
