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Consumption of pollen contaminated with field-realistic concentrations of fungicide cause sub-lethal effects in common eastern bumble bee (Bombus impatiens [Hymenoptera]: [Apidae]) microcolonies

Runnion, Emily1; Strage, James1; Sivakoff, Frances2

Published May 06, 2024 on Dryad. https://doi.org/10.5061/dryad.rv15dv4f4

Cite this dataset

Runnion, Emily; Strage, James; Sivakoff, Frances (2024). Consumption of pollen contaminated with field-realistic concentrations of fungicide cause sub-lethal effects in common eastern bumble bee (Bombus impatiens [Hymenoptera]: [Apidae]) microcolonies [Dataset]. Dryad. https://doi.org/10.5061/dryad.rv15dv4f4

Abstract

Bumble bees are declining across the globe. Causes of this decline have been attributed to a variety of stressors, including pesticides. Fungicides are a type of pesticide that have been understudied in the context of bumble bee health. As a result, fungicides are often applied to flowering plants without consideration of pollinator exposure. Recent work demonstrates that fungicides have sublethal effects in bumble bees, but little is known about how much fungicide it takes to cause these sublethal effects. To address this gap in the literature, we fed microcolonies of the common eastern bumble bee (Bombus impatiens) pollen contaminated with a range of fungicide concentrations. We chose these concentrations based on the range of fungicide concentrations in pollen and nectar that were reported in the literature. Results revealed that later-stage pupae and newly emerged males are potentially sensitive to fungicide exposure, showing smaller size and reduced fat reserves at intermediate levels of contamination. Compared to the control, intermediated levels of fungicide-contaminated pollen led to increased pupal mortality and delayed male emergence. Contrary to expectations, higher fungicide levels did not exhibit a linear relationship with negative impacts, suggesting nuanced effects. Because body size and emergence timing are important aspects of bumble bee reproductive behavior, results have implications for mating success, potentially disrupting colony development.

README

This README file was generated on 2024-05-02 by Emily Runnion.

GENERAL INFORMATION

  1. Title of Dataset: Consumption of pollen contaminated with field-realistic concentrations of fungicide cause sub-lethal effects in common eastern bumble bee (Bombus impatiens [Hymenoptera]: [Apidae]) microcolonies

  2. Author Information
    A. Principal Investigator Contact Information
    Name: Emily Runnion
    Institution: The Ohio State University
    Address: Columbus, OH USA
    Email: Runnion.4@osu.edu

    B. Associate or Co-investigator Contact Information
    Name: Jamie Strange
    Institution: The Ohio State university
    Address: Columbus, OH USA
    Name: Frances Sivakoff
    Institution: The Ohio State university
    Address: Columbus, OH USA

  3. Date of data collection (single date, range, approximate date): 2022

  4. Geographic location of data collection: Columbus, Ohio, USA

  5. Information about funding sources that supported the collection of the data: This work was funded by Sustainable Agriculture Research and Education Projects, North-Central Region, Grant Agreement GNC21-335 (to ENR) and an OSU CFAES Research & Graduate Education Internal Grant (to FSS and JPS).

  6. Licenses/restrictions placed on the data: CC0 1.0 Universal (CC0 1.0) Public Domain

  7. Links to publications that cite or use the data:

  8. Links to other publicly accessible locations of the data: None

  9. Links/relationships to ancillary data sets: None

  10. Was data derived from another source? No
    A. If yes, list source(s): NA

DATA & FILE OVERVIEW

  1. File List:

A) brood.csv
C) drone.health.csv
D) pollen.csv
E) qro.csv
F) workers.csv

  1. Relationship between files, if important: None
  2. Additional related data collected that was not included in the current data package: None
  3. Are there multiple versions of the dataset? No A. If yes, name of file(s) that was updated: NA i. Why was the file updated? NA ii. When was the file updated? NA

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DATA-SPECIFIC INFORMATION FOR: brood.csv

  1. Number of variables: 24
  2. Number of cases/rows: 145
  3. Variable List:
  • colony: unique identifier for each of the 45 microcolonies
  • whole.mean: the average amount of pollen eaten, per unique microcolony, every 24 hours from beginning to end of the experiment
  • mean.dose - the average amount of Pristine (fungicide) consumed per unique microcolony, every 24 hours from beginning to end of the experiment
  • dose: the amount of Pristine (fungicide) added in ppb to each 1 gram of pollen fed to each microcolony every 24 hours
  • treatment: Number 1-5, signifying the concentration of Pristine (fungicide) fed to each microcolony. Treatment 1 = Control, no fungicide. Treatment 2 = 150ppb, Treatment 3 = 1500ppb, Treatment 4 = 15000ppb, Treatment 5 = 150000ppb
  • replicate: blocking variable
  • brood_cells = the total count of pupae, larvae, egg sacs, and honey pots per microcolony at the conclusion of the experiment, following microcolony dissection
  • honey_pot = total count of honey pots per microcolony. Honey pot determined as a open pula cell containing nectar.
  • eggs = total count of eggs per microcolony at the end of the experiment following microcolony dissection
  • dead_larvae: total count of dead larvae per microcolony at the end of the experiment following microcolony dissection
  • live_larvae: total count of live larvae per microcolony at the end of the experiment following microcolony dissection
  • dead_pupae: total count of dead pupae per microcolony at the end of the experiment following microcolony dissection
  • live_pupae: total count of live pupae per microcolony at the end of the experiment following microcolony dissection
  • dead_drones: total count of dead drones per microcolony at the end of the experiment following microcolony dissection. dead_drones were counted as those males that did not successfully emerge, but surpassed the pupal stage, as seen by removal of the brood cell capping, but due to deformed wings, unknown illness, etc., were not able to exit the brood cell.
  • live_drones: total count of males found outside of brood cells within post-frozen microcolonies and subsequent dissection.
  • drones: total count of successfully emerged new male bees per microcolony over the course of the experiment.
  • qro: unique identifier given to the original queenright colony which the workers in the matching microcolony were sourced from.
  • duration: amount of time the microcolony had living workers, from the day callow workers were placed from the original queenright colony until the day the microcolony was frozen. Microcolonies were frozen either 1) when all workers died, 2) seven days after the first male emerged, or 3) within two standard deviations of the average microcolony duration (this was used for microcolonies that did not produce males)
  • alive: the number of workers alive inside the colony when the microcolony was frozen
  • dead: the number of workers out of 5 who were dead when the microcolony was frozen
  1. Missing data codes: None
  2. Specialized formats or other abbreviations used: None

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DATA-SPECIFIC INFORMATION FOR: drone.health.csv

  1. Number of variables: 21
  2. Number of cases/rows: 421
  3. Variable List:
  • colony: unique identifier for each of the 45 microcolonies
  • whole.mean: the average amount of pollen eaten, per unique microcolony, every 24 hours from beginning to end of the experiment
  • mean.dose - the average amount of Pristine (fungicide) consumed per unique microcolony, every 24 hours from beginning to end of the experiment
  • dose: the amount of Pristine (fungicide) added in ppb to each 1 gram of pollen fed to each microcolony every 24 hours
  • treatment: Number 1-5, signifying the concentration of Pristine (fungicide) fed to each microcolony. Treatment 1 = Control, no fungicide. Treatment 2 = 150ppb, Treatment 3 = 1500ppb, Treatment 4 = 15000ppb, Treatment 5 = 150000ppb
  • replicate: blocking variable
  • id - unique identifier given to each emerged male
  • emerge_time: time since the initiation of the microcolony until this row's male emerged
  • wet_weight: the weight of the newly emerged drone, in grams, while alive
  • dry_weight: the weight of the whole body of the male after freezing, and then being dried in an oven at 60 degrees Celsius for 72 hours
  • radial: the length of the male's right radial wing vein, in micrometers
  • abdomen_dry: the weight, in grams, of the dried abdomen of the male. Abdomens removed immediately after recording whole-body dry weight.
  • order_on_sheet: order in which the males emerged during the experiment
  • abdomen_post_ethyl: the weight of the male's abdomen, in grams, after being removed following the initial 72 hour drying, soaked in ethyl ether for 24 hours, and dried again. See Methods for particulars.
  • fat_content: the fat content of the male's abdomen, calculated as the difference between the "abdomen_dry" value and the "abdomen_post_ethyl" value
  • relative_fat: the relative fat content of the male's abdomen, calculated as the value of "abdomen_post_ethyl" divided by "radial"
  • workers_alive: the number of workers alive inside the colony when the male emerged
  • qro: unique identifier given to the original queenright colony which the workers in the matching microcolony were sourced from.
  • duration: amount of time the microcolony had living workers, from the day callow workers were placed from the original queenright colony until the day the microcolony was frozen. Microcolonies were frozen either 1) when all workers died, 2) seven days after the first male emerged, or 3) within two standard deviations of the average microcolony duration (this was used for microcolonies that did not produce males)
  1. Missing data codes: NA (data not available)

Missing data described based on male "id"

1.11R2.1
3.7R2.3
3.9R2.2
3.7R2.2
4.11R2.1
4.4R2.7
1.4R2.2
4.4R2.8
1.4R2.3
These males had deformed wings and we could not measure the radial cell length

4.12R2.2
2.1R2.7
4.3R2.2
1.5R2.1
3.3R2.3
4.4R2.5
1.7R2.1
1.4R2.1
4.3R2.3
4.4R2.9
4.4R2.3
4.4R2.1
4.3R2.5
3.3R2.2
4.4R2.4
4.3R2.1
4.3R2.4
5.5R2.1
3.3R2.1
4.4R2.6
1.7R2.2
4.4R2.2
These males' abdomens were damaged during the ethyl ether process

5.12R2.1
4.11R2.2
We forgot to record these male's wet weight

4.4R2.13
5.7R2.5
1.5R2.6
These males' relative fat values were negative and therefore not included

  1. Specialized formats or other abbreviations used: None

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DATA-SPECIFIC INFORMATION FOR: pollen.csv

These are the data used to calculate relative body mass, then analyze change in relative body mass for recaptures (i.e., birds captured in the pre and post periods of the experiment).

  1. Number of variables: 18
  2. Number of cases/rows: 933
  3. Variable List:
  • colony: unique identifier for each of the 45 microcolonies
  • treatment: Number 1-5, signifying the concentration of Pristine (fungicide) fed to each microcolony. Treatment 1 = Control, no fungicide. Treatment 2 = 150ppb, Treatment 3 = 1500ppb, Treatment 4 = 15000ppb, Treatment 5 = 150000ppb
  • dose: the amount of Pristine (fungicide) added in ppb to each 1 gram of pollen fed to each microcolony every 24 hours \
  • pristine/g: the amount of Pristine (Fungicide) per gram in each pollen ball
  • replicate: blocking variable
  • count: the pollen ball count per microcolony. For example, "4" would mean this was the 4th pollen ball supplied to the microcolony.
  • start_date: the day the pollen ball was placed in the microcolony
  • start_time: the time the pollen ball was placed in the microcolony
  • start_weight: the weight, in grams, of the pollen ball when placed in the microcolony
  • end_date: the day the pollen ball was removed from the microcolony
  • end_time: the time the pollen ball was removed from the microcolony
  • end_weight: the weight of the pollen ball, in grams, after being removed from the microcolony following a period of 48 hours of allowing the workers free access to eat the pollen
  • whole_dif: the raw difference between "start_weight" and "end_weight" in grams
  • difference: the weight difference between the "start_weight" and "end_weight" values after adding back in the weight calculated lost from moisture using the amount of moisture lost from the evaporative control pollen ball assigned to that microcolony that week
  • percent_consumed: the percent of the pollen ball consumed by the microcolony ("end_weight" divided by "start_weight")
  • dose_consumed: the calculated amount of Pristine (fungicide) consumed by the microcolony from that pollen ball, in Pristine/g
  • pid: unique identifier for each individual pollen ball
  • bees_alive: the amount of workers alive in the colony at the time the "end_weight" was recorded
  1. Missing data codes: NA (data not applicable)

Missing data described based on "pid"

R2.3.7-21
This pollen ball was not removed from the box because there were eggs in it

R2.1.9-17
R2.3.11-5
R2.4.12-4
R2.5.12-12
These pollen balls were not included in analysis because they contained too much frass to be adequately measured

  1. Specialized formats or other abbreviations used: None

#########################################################################

DATA-SPECIFIC INFORMATION FOR: qro.csv

  1. Number of variables: 2
  2. Number of cases/rows: 45
  3. Variable List:
  • colony: unique identifier for each of the 45 microcolonies
  • qro: unique identifier given to the original queenright colony which the workers in the matching microcolony were sourced from.
  1. Missing data codes: None
  2. Specialized formats or other abbreviations used: None

#########################################################################

DATA-SPECIFIC INFORMATION FOR: workers.csv

  1. Number of variables: 17
  2. Number of cases/rows: 224
  3. Variable List:
  • colony: unique identifier for each of the 45 microcolonies
  • whole.mean: the average amount of pollen eaten, per unique microcolony, every 24 hours from beginning to end of the experiment
  • mean.dose - the average amount of Pristine (fungicide) consumed per unique microcolony, every 24 hours from beginning to end of the experiment
  • treatment: Number 1-5, signifying the concentration of Pristine (fungicide) fed to each microcolony. Treatment 1 = Control, no fungicide. Treatment 2 = 150ppb, Treatment 3 = 1500ppb, Treatment 4 = 15000ppb, Treatment 5 = 150000ppb
  • replicate: blocking variable
  • start_wet_weight: the weight of the live worker in grams on the day they were first placed in the microcolony
  • alive_at_end: a logical variable, coded "1" for yes/true and "0" for no/false, to signify if the worker bee was alive when the microcolony was shut down
  • shut_down_dt: the day the microcolony was placed in the freezer
  • dry_weight: the whole body dry weight of the worker bee in grams after being frozen and dried for 72 hours at 60 degrees Celsius
  • days_alive: the number of the days the worker bee was alive during the duration of the experiment
  • colony_duration: amount of time the microcolony had living workers, from the day callow workers were placed from the original queenright colony until the day the microcolony was frozen. Microcolonies were frozen either 1) when all workers died, 2) seven days after the first male emerged, or 3) within two standard deviations of the average microcolony duration (this was used for microcolonies that did not produce males)
  • qro: unique identifier given to the original queenright colony which the workers in the matching microcolony were sourced from.
  • alive: the number of workers alive inside the colony when the microcolony was frozen
  • dead: the number of workers out of 5 who were dead when the microcolony was frozen
  1. Missing data codes: none
  2. Specialized formats or other abbreviations used: None

#########################################################################

Methods

To understand how fungicides affect bumble bee microcolony health and reproduction, we established five fungicide treatments, which varied by the concentration of Pristine® in their pollen diet. We selected experimental values of Pristine® of 0 ppb, 150 ppb, 1,500 ppb, 15,000 ppb and 150,000 ppb.We exposed bumble bees to fungicide by incorporating it into pollen balls, which we made in batches per treatment by mixing 90g of honey bee-collected pollen with 35ml of a 50/50 sucrose:water solution containing the appropriate concentration of fungicide. We made pollen balls for each treatment in separate batches, from a single supply of homogenized honey bee-collected pollen. Using callow worker bees from four purchased B. impatiens queen-right colonies (Plant Products USA, Westland, MI), we established microcolonies consisting of five individuals, supplied with a 50/50 sucrose:water solution, a 1 g (±0.5 g) fungicide-free pollen ball, and 1-2 g of wax material from their natal colony to stimulate oviposition and brood production behavior. We replicated each of the five fungicide treatments nine times, for a total of 45 microcolonies. We placed microcolonies in a rearing room under red light at The Ohio State University’s Rothenbuhler Honey Bee Research Laboratory. Rearing room conditions were kept between 30°C-32°C, and 50%-70% relative humidity. We blocked by location within the rearing room, where each block contained microcolonies from each fungicide treatment and ensured that all workers within a block originated from the same original queenright colony to control for potential confounding genetic differences. Our experiment ran from August to October 2022, during which time we monitored the pollen consumption, brood production, and mortality in the 45 microcolonies. We stored data in Microsoft Excel, and ran all analyses in R. 

Funding

Sustainable Agriculture Research and Education Projects, Award: GNC21-335

OSU CFAES Research & Graduate Education Internal Grant, Award: GR120070