DOI: 10.5061/dryad.kwh70rzcm
title: Preventative medicine? Examining prophylactic effects of a sunflower pollen diet in Bombus impatiens

description: The widespread decline of pollinator populations is of concern for both
 natural and agricultural ecosystems. Pathogens have been identified as a
 major contributor to the decline of some bee species, making understanding
 host-pathogen dynamics a crucial area of research. Sunflower pollen
 (Helianthus annuus) dramatically and consistently reduces infection by a
 prevalent gut pathogen, Crithidia bombi, in the common eastern bumble bee
 (Bombus impatiens), when consumed by bees post-infection, but we do not
 know if sunflower can confer protection when consumed before exposure. We
 asked whether feeding bumble bees sunflower pollen diets before pathogen
 exposure decreases Crithidia infection compared to buckwheat pollen
 (Fagopyrum esculentum). Buckwheat pollen was used as a comparison since it
 has a similar protein concentration as sunflower pollen, but results in
 high Crithidia counts when consumed post-infection. Bumble bees were fed
 sunflower or buckwheat pollen for seven days, inoculated with Crithidia,
 and then fed a wildflower pollen control diet for seven more days before
 assessing infection. We found that consuming a sunflower pollen diet
 before inoculation did not reduce Crithidia cell counts compared to
 buckwheat pollen. Further, bumble bee survival and consumption of sucrose
 solution and pollen did not differ between these diets. The results show
 no evidence of sunflower pollen providing prophylactic resistance against
 Crithidia bombi infection, indicating that the timing at which sunflower
 pollen is consumed relative to exposure has important consequences for
 infection.
description: These data are from an experimental manipulative study performed
 in a lab setting to ask whether exposure to specific diets provides
 prophylactic protection against infection by a gut pathogen. Commercial
 bumble bees (<em>Bombus impatiens</em>) were assigned to be
 fed sunflower (<em>Helianthus annuus</em>) or buckwheat
 (<em>Fagopyrum esculentum</em>) pollen for one week and were
 then inoculated with a standard dose of <em>Crithidia
 bombi</em>, a gut pathogen. After inoculation, all bees were fed a
 standard wildflower pollen diet for one more week and then assessed to
 determine infection intensity (<em>C. bombi</em> cells per
 0.02 uL).
description: # Preventative medicine? Examining prophylactic effects of a sunflower
 pollen diet in *Bombus impatiens*
 [https://doi.org/10.5061/dryad.kwh70rzcm](https://doi.org/10.5061/dryad.kwh70rzcm) This file describes the archived R script and CSV files associated with this manuscript. ## Code/Software: R script: ProphylacticScript-CL\_Final.R All the analyses and graphs in the manuscript are produced by this R script, which includes a "set working directory" command within the code for each question, which you will need to modify. The script is written so that it works if all CSV data files are put into one folder, and the working directory is set to that folder. This has only been tested under Windows but should also be true in macOS and Linux. ## Description of the data and file structure These CSV files were extracted from original data files in XLS or XLSX format. Missing data are indicated with 'NA'. The CSV files are included in the archive so that the R script will run "out of the box" to replicate analyses in the paper. All dates are in month/day/year format. ### **Files**: #### **MasterDataSunflowerProphylaxis\_CL\_ECM\_CountData.csv** Sample\_ ID: Bee individual identification number Treatment: Diet treatment pollen the bee was given (either sunflower or buckwheat) Colony_ID: Source colony name Inoc_Date: Date the bee was inoculated with Crithidia Dissect_Date: Date the bee was dissected Count: Crithidia cell count in 0.02 uL bee gut solution (hindgut macerated in 300 uL Ringer's solution) Death: 1 = died before dissection, 0 = did not die before dissection Death_Date: If the bee died before dissection, the date of death Survival_days: Days survived for bees who died before dissection wing_cm: Marginal cell length in cm of the right wing wing_mm: Marginal cell length in mm of the right wing #### **MasterSunflowerConsumption\_CL\_Final.csv** ID: Bee identification number treatment: Diet treatment pollen the bee was given (SF = sunflower, BW = buckwheat) pol_bef_date: Date that treatment pollen weight was measured before the bee was allowed to consume treatment pollen pol_bef_time: Time that treatment pollen weight was measured before the bee was allowed to consume treatment pollen pol_bef_weight_g: Weight in grams of treatment pollen before the bee was allowed to consume treatment pollen pol_aft_date: Date that treatment pollen weight was measured after the bee was allowed to consume treatment pollen pol_aft_time: The time that treatment pollen weight was measured after the bee was allowed to consume treatment pollen pol_aft_weight_g: Weight in grams of treatment pollen after the bee was allowed to consume treatment pollen suc_bef_date: Date that sucrose weight was measured before the bee was allowed to consume treatment pollen suc_bef_time: Time that sucrose weight was measured before the bee was allowed to consume treatment pollen suc_bef_weight_g: Weight in grams of sucrose before the bee was allowed to consume treatment pollen suc_aft_date: Date that sucrose weight was measured bee was allowed to consume treatment pollen suc_aft_Time: Time that sucrose weight was measured and the bee was allowed to consume treatment pollen suc_aft_weight_g: Weight in grams of sucrose after the bee was allowed to consume treatment pollen Net_pollen: Change in pollen weight between pol_bef_weight_g and pol_aft_weight_g Net_sucrose: Change in sucrose weight between suc_bef_weight_g and suc_aft_weight_g #### **MasterSunflowerEvaporation_CL_Final.csv** ID: Evaporative control identification. C indicates evaporation control, the number differentiates between evaporation controls treatment: Diet treatment pollen used in the evaporation control (SF = sunflower, BW = buckwheat) pol_bef_date: Date that treatment pollen weight was measured before the evaporation control period pol_bef_time: Time that treatment pollen weight was measured before the evaporation control period pol_bef_weight_g: Weight in grams of treatment pollen before the evaporation control period pol_aft_date: Date that treatment pollen weight was measured after the evaporation control period pol_aft_time: The time that treatment pollen weight was measured after the evaporation control period pol_aft_weight_g: Weight in grams of treatment pollen after the evaporation control period suc_bef_date: Date that sucrose weight was measured before the evaporation control period suc_bef_time: Time that sucrose weight was measured before the evaporation control period suc_bef_weight_g: Weight in grams of sucrose before the evaporation control period suc_aft_date: Date that sucrose weight was measured after the evaporation control period suc_aft_Time: The time that sucrose weight was measured after the evaporation control period suc_aft_weight_g: Weight in grams of sucrose after the evaporation control period Net_pollen: Change in pollen weight between pol_bef_weight_g and pol_aft_weight_g Net_sucrose: Change in sucrose weight between suc_bef_weight_g and suc_aft_weight_g