DOI: 10.5061/dryad.wpzgmsc08
title: A nutritionally complete pollen-replacing diet protects honey bee colonies during stressful commercial pollination: Requirement for isofucosterol

description: A steady supply of nutritionally adequate pollen from diverse flower
 sources is crucial for honeybee colonies. However, climate instability,
 large-scale agriculture, and the loss of flower-rich landscapes have made
 this supply scarce and unpredictable, threatening both apiculture and
 sustainable crop pollination. We developed a nutritionally
 complete pollen-replacing diet that supports continuous brood production
 from May to October in colonies without access to pollen. Omitting
 isofucosterol, the third most abundant sterol in honeybees, causes
 significant reductions in brood production and neuromuscular dysfunction
 in adults, identifying isofucosterol as a critical micronutrient. In
 contrast, omitting 24-methylene cholesterol, the most abundant honeybee
 sterol, does not significantly affect brood production, and surprisingly,
 bees remain viable without it. Colonies fed a commercial diet severely
 declined in brood production after 36 days and died out. In a season-long
 experiment in the commercial pollination of blueberry and sunflower
 fields, a treatment group fed the complete diet overcame the detrimental
 effects of nutritional stress, unlike colonies in "No Diet" and
 "Commercial Diet" groups. This study suggests that
 feeding a complete, pollen-replacing diet to nutritionally stressed
 colonies can address the root causes of honeybees' growing
 nutritional deficiencies, supporting their health and their vital
 pollination services.
description: # A nutritionally complete pollen-replacing diet protects honey bee
 colonies during stressful commercial pollination: Requirement for
 isofucosterol Dataset DOI:
 [https://doi.org/10.5061/dryad.wpzgmsc08](https://doi.org/10.5061/dryad.wpzgmsc08) ## Description of the data and file structure ### Data Files: #### 1. "Tent_Wingene_Manuscript.csv" **Description:** This file contains experimental data on the tent study (Experiment 1) with no access to external food. Data was collected in Wingene, Belgium. Capped brood data was collected from Day 24, being 16/06/202,3 until Day 108, being 08/09/2023. **Variables:** - `ID` (integer): Unique identifier for the hive. - `Tent` (integer): Number of the tent. - `TRT` (string): Name of the diet. - `Day` (string): Day of assessment - `Brood` (integer, count): Capped brood cells produced in a colony. - `Brood_CumD24` (integer, cumulative count): Cumulative count of capped brood cells. #### 2. "Pollination_trial.csv" **Description:** This file contains experimental data on the impact of Diet A on nutritionally stressed colonies under field conditions. Data was collected in North-Western Washington State near the town of Mount Vernon, WA, USA, and Eastern Washington State near the town of Othello, USA. The experiment started on May 12th, 2023 (Day 0). **Variables:** - `ID` (integer): Unique identifier for the hive. - `TRT` (string): Name of the diet. - `Assessment` (string): Day of assessment, A1 = Day 11, A2 = Day 25, A3 = Day 41, A4 = Day 68, A5 = Day 106, A6 = Day 133 - `Bees` (numeric): frames of bees - `Brood` (numeric): Number of 5 cm^2 grid squares covered with capped brood ## R Script Outputs: All analyses were performed in the R version 4.3.1 [R Core Team. R: A language and environment for statistical computing. [Online] 2023. [https://www.R-project.org](https://www.R-project.org)] using the lme4 package [Bates D. 2015 Fitting linear mixed-effects models using lme4. J. Stat. Softw. 67, 48]. The code is included in the submission, including detailed comments on the loaded packages used and a step-by-step description. #### 1. "Tent_Trial_Manuscript.R" **Description:** This script processes and analyzes the experimental data (`Tent_Wingene_Manuscript.csv`). It performs statistical tests to compare treatment effects on counts and cumulative counts of capped brood cells throughout the experiment. **Outputs:** - Visualisations on the changes in count of capped brood cells (Supplemental Material Figure 4) and changes in cumulative capped brood cells (Supplemental Material Figure 5). - Linear Mixed Effects Model (LMEM) for the change in count of capper brood cells from day 24 to day 108 (Supplemental Material Table 5) and for the change in cumulative count of capped brood cells from day 24 to day 108 (Supplemental Material Table 6). #### 2. "Pollination_trial_Manuscript.R" **Description:** This script processes and analyzes the experimental data (`Pollination_trial_Manuscript.csv`). It performs statistical tests to compare treatment effects on frames of bees and counts of capped brood cells over the course of the experiment. **Outputs:** - Visualisations on the changes in frames of bees (Supplemental Material Figure 9) and changes in the count of capped brood cells (Supplemental Material Figure 11). - Linear Mixed Effects Model (LMEM) for the change in for change in count of frames of bees (Supplemental Material Table 9) and for the change in number of 5 cm^2 squares with capped brood (Supplemental Material Table 10). #### "Supplemental material Final.pdf" **Description:** Contains all supplemental materials, including figures, tables, and statistics. ## Access information NA