Location: Honey Bee Research
Project Number: 5342-21000-017-00
Start Date: Feb 07, 2014
End Date: Feb 06, 2019
Sub-objective 1A: Determine whether beebread contains a core microbial community. Beebread will be sampled from multiple colonies, apiaries, and commercial operations across a variety of locations and seasons. rRNA will be used to characterize the active microbiota of beebread. Fungal and bacterial groups identified at different levels of taxonomic certainty will be examined for significant co-occurrence using a variety of available metrics (including options for degenerate matrices) and a null hypothesis of random community assembly. Sub-objective 2A: Determine whether the active microbial (bacterial and fungal) community remains constant as corbicular pollen becomes beebread and as beebread ages. We will detail the active fungal and bacterial communities in multiple colonies, controlling for the source of corbicular pollen and season. Multiple replicates of beebread at 0.5, 1, 3, 7, 14, and 30 days of age will be sampled and processed for microbial composition. Dependent upon the predictability of a "core" functional set of successional genes in beebread, we will develop metagenomic profiling methods for a more efficient characterization of microbial function. Sub-objective 2B: Determine whether the microbial communities of overwintered beebread sampled from old wax comb differ from those of new wax comb. Beebread will be collected in RNA later from both old and new wax comb from commercial beekeeping operations. RNA extracted from beebread will be subject to 454 amplicon sequencing and compared according to overwintering status and comb age. If differences in the microbial communities are discovered, we will determine the nutrients, preservatives and metabolites associated with these changes. Sub-objective 3A: Determine whether supplemental feed affects the active honey bee gut microbial community. Bees will be fed commonly used brewer's yeast/soy/sucrose-based nutritional supplements containing thymol alone, citric acid alone, thymol and citric acid, honey bee healthy, no additives, and fresh beebread/honey as a control. RNA will be extracted from these pooled samples and community composition of the gut will be assessed using qRT-PCR targeted to the core gut bacteria. Sub-objective 3B: Determine whether the active microbial communities in beebread differ by pollination environment. We will sample both corbicular pollen and beebread microbial communities of colonies actively pollinating two distantly located monocultures and two distantly located plant polycultures. Samples will be subject to qRT-PCR targeting specific genera, and also pooled by colony and subject to 454 amplicon sequencing for comparative purposes. If species-specific qRT-PCR primers prove overly time consuming or inefficient, we will rely on the sequencing of major functional COGs. Sub-objective 3C: Determine whether the microbes and nutrition in bee bread are affected by fungicide. Endura fungicide will be applied at field concentrations to a Brassica mix grown in greenhouses. Bees will be allowed to forage on fungicide sprayed and non-fungicide controls. Beebread will be examined for microbial communities, fungicide concentrations and nutritional analysis.