Abstract
Many pollinator species are declining due to a variety of interacting stressors including pathogens, sparking interest in understanding factors that could mitigate these outcomes. Diet can affect host-pathogen interactions by changing nutritional reserves or providing bioactive secondary chemicals. Recent work found that sunflower pollen (Helianthus annuus) dramatically reduced cell counts of the gut pathogen Crithidia bombi in bumble bee workers (Bombus impatiens), but the mechanism underlying this effect is unknown. Here we analyzed methanolic extracts of sunflower pollen by LC-MS and identified triscoumaroyl spermidines as the major secondary metabolite components, along with a flavonoid quercetin-3-O-hexoside and a quercetin-3-O-(6-O-malonyl)-hexoside. We then tested the effect of triscoumaroyl spermidine and rutin (as a proxy for quercetin glycosides) on Crithidia infection in B. impatiens, compared to buckwheat pollen (Fagopyrum esculentum) as a negative control and sunflower pollen as a positive control. In addition, we tested the effect of nine fatty acids from sunflower pollen individually and in combination using similar methods. Although sunflower pollen consistently reduced Crithidia relative to control pollen, none of the compounds we tested had significant effects. In addition, diet treatments did not affect mortality, or sucrose or pollen consumption. Thus, the mechanisms underlying the medicinal effect of sunflower are still unknown; future work could use bioactivity-guided fractionation to more efficiently target compounds of interest, and explore non-chemical mechanisms. Ultimately, identifying the mechanism underlying the effect of sunflower pollen on pathogens will open up new avenues for managing bee health.
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Acknowledgements
We thank E. Amponsah, L. Cleary, J. Cook, L. Gagnon, Z. Henry, K. Michaud, J. Roch, and C. Sylvia for assistance with experiments, and two anonymous reviewers for feedback on the manuscript. Funding came from USDA-AFRI 2013-02536 (LSA, REI and PCS), USDA-NIFA-2016-07962 and USDA-NIFA-2018-08591 (both to LSA and REI), the USDA/CSREES (Multi-state Hatch) MAS00497 (LSA), and the Peter Sowerby Foundation in the UK (PCS).
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LSA, REI and PCS conceived of and designed the study. PCS and IWF conducted chemical analysis of pollen and synthesized spermidines. AEF, RLM, PRA, LMC, PMD, and SL carried out bioassay experiments. AEF analyzed data and prepared figures. LSA wrote the manuscript with substantial contributions from PCS. All co-authors read and provided feedback on the manuscript.
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Adler, L.S., Fowler, A.E., Malfi, R.L. et al. Assessing Chemical Mechanisms Underlying the Effects of Sunflower Pollen on a Gut Pathogen in Bumble Bees. J Chem Ecol 46, 649–658 (2020). https://doi.org/10.1007/s10886-020-01168-4
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DOI: https://doi.org/10.1007/s10886-020-01168-4