Land conversion and pesticide use degrade forage areas for honey bees in America's beekeeping epicenter.

A diverse range of threats have been associated with managed-bee declines globally. Recent increases of two known threats, land-use change and pesticide use, have resulted from agricultural expansion and intensification notably in the top honey-producing state in the United States: North Dakota. This study investigated the dual threat from land conversion and pesticide use surrounding ~14,000 registered apiaries in North Dakota from 2001 to 2014. We estimated the annual total insecticide use (kg) on major crops within 1.6 km of apiary sites. Of the eight insecticides quantified, six showed significant increasing trends over the time period. Specifically, applications of the newly established neonicotinoids Chlothianidin, Imidacloprid and Thiamethoxam, increased annually by 1329 kg, 686 kg, 795 kg, respectively. Also, the use of Chlorpyrifos, which was well-established in the state by 2001 and is highly toxic to honey bees, increased by ~8,800 kg annually from 6,500 kg in 2001 to 115,000 kg in 2014 on corn, soybeans and wheat. We further evaluated the relative quality changes of natural/semi-natural land covers surrounding apiaries in 2006, 2010 and 2014, a period of significant increases in cropland area. In areas surrounding apiaries, we observed changes in multiple indices of forage quality that reflect the deteriorating landscape surrounding registered apiary sites due to land-use change and pesticide-use increases. Overall, our results suggest that the application of foliar-applied insecticides, including pyrethroids and one organophosphate, increased surrounding apiaries when the use of neonicotinoid seed treatments surged and the area for producing corn and soybeans expanded. Spatially, these threats were most pronounced in southeastern North Dakota, a region hosting a high density of apiary sites that has recently experienced corn and soybean expansion. Our results highlight the value of natural and semi-natural land covers as sources of pollinator forage and refugia for bees against pesticide exposure. Our study provides insights for targeting conservation efforts to improve forage quality benefiting managed pollinators.

Do the Quality and Quantity of Honey Bee-Collected Pollen Vary Across an Agricultural Land-Use Gradient?

Pollen is the source of protein for most bee species, yet the quality and quantity of pollen is variable across landscapes and growing seasons. Understanding the role of landscapes in providing nutritious forage to bees is important for pollinator health, particularly in areas undergoing significant land-use change such as in the Northern Great Plains (NGP) region of the United States where grasslands are being converted to row crops. We investigated how the quality and quantity of pollen collected by honey bees (Apis mellifera L. [Hymenoptera: Apidae]) changed with land use and across the growing season by sampling bee-collected pollen from apiaries in North Dakota, South Dakota, and Minnesota, USA, throughout the flowering season in 2015-2016. We quantified protein content and quantity of pollen to investigate how they varied temporally and across a land-use gradient of grasslands to row crops. Neither pollen weight nor crude protein content varied linearly across the land-use gradient; however, there were significant interactions between land use and sampling date across the season, particularly in grasslands. Generally, pollen protein peaked mid-July while pollen weight had two maxima in late-June and late-August. Results suggest that while land use itself may not correlate with the quality or quantity of pollen resources collected by honey bees among our study apiaries, the nutritional landscape of the NGP is seasonally dynamic, especially in certain land covers, and may impose seasonal resource limitations for both managed and native bee species. Furthermore, results indicate periods of qualitative and quantitative pollen dearth may not coincide.

Taxonomic Characterization of Honey Bee (Apis mellifera) Pollen Foraging Based on Non-Overlapping Paired-End Sequencing of Nuclear Ribosomal Loci.

Identifying plant taxa that honey bees (Apis mellifera) forage upon is of great apicultural interest, but traditional methods are labor intensive and may lack resolution. Here we evaluate a high-throughput genetic barcoding approach to characterize trap-collected pollen from multiple North Dakota apiaries across multiple years. We used the Illumina MiSeq platform to generate sequence scaffolds from non-overlapping 300-bp paired-end sequencing reads of the ribosomal internal transcribed spacers (ITS). Full-length sequence scaffolds represented ~530 bp of ITS sequence after adapter trimming, drawn from the 5' of ITS1 and the 3' of ITS2, while skipping the uninformative 5.8S region. Operational taxonomic units (OTUs) were picked from scaffolds clustered at 97% identity, searched by BLAST against the nt database, and given taxonomic assignments using the paired-read lowest common ancestor approach. Taxonomic assignments and quantitative patterns were consistent with known plant distributions, phenology, and observational reports of pollen foraging, but revealed an unexpected contribution from non-crop graminoids and wetland plants. The mean number of plant species assignments per sample was 23.0 (+/- 5.5) and the mean species diversity (effective number of equally abundant species) was 3.3 (+/- 1.2). Bray-Curtis similarities showed good agreement among samples from the same apiary and sampling date. Rarefaction plots indicated that fewer than 50,000 reads are typically needed to characterize pollen samples of this complexity. Our results show that a pre-compiled, curated reference database is not essential for genus-level assignments, but species-level assignments are hindered by database gaps, reference length variation, and probable errors in the taxonomic assignment, requiring post-hoc evaluation. Although the effective per-sample yield achieved using custom MiSeq amplicon primers was less than the machine maximum, primarily due to lower "read2" quality, further protocol optimization and/or a modest reduction in multiplex scale should offset this difficulty. As small quantities of pollen are sufficient for amplification, our approach might be extendable to other questions or species for which large pollen samples are not available.