Toxic temperatures: bee behaviours exhibit divergent pesticide toxicity relationships with warming Kenna, Graystock & Gill Article doi: 10.1111/gcb.16671
Data files representing quantitative data on bee behavioural responses to temperature and pesticide combination treatments for all three experiments. Includes: csv files for statistical analyses; Excel files as further supporting data; R scripts showing the code for data wrangling, visualisation, and stat model fitting; MKV file showing bee movement.
Experiment 1 files
Controlled Environment (CE) room conditions File names(s): ‘CE_Room_C_exp1’; ‘CE_Room_E_exp1’; ‘CE_Room_G_exp1’ CSV files providing humidity and temperature logger data evet minute (each row) over the period the experiment was run, showing high consistency. Continuous response variable. Variables Humidity = Relative humidity in % Temp = Room temperarure in degrees Celsius
Body size File names(s): ‘Size_Data_exp1’ CSV file providing the intertegula distance for each tested bee. Continuous response variable. Variables Temp = temperarure treatment (21[low]; 27[mid]; 30[high] Pesticide = C=control; I= imidacloprid; S=sulfoxaflor Colony = Number of the natal donor colony ITS = InterTegula Span in millimeters
Responsiveness File names(s): ‘Incapacitated12hr’; ‘Death12hr’ CSV files showing if bees responded to the stimulus or were found dead, respectively. Binary response variable. Variables Pot - Number of holding cage Colony = Number of the natal donor colony Temperature = temperarure treatment (21[low]; 27[mid]; 30[high] Pesticide = control; imidacloprid; sulfoxaflor ID = Indivdiual identification code for each worker Columns G [0] - S [144) is the number of hours since onset of treatment exposure = A=Active; I=Incapacitated; hyphen=Dead
Feeding File names(s): ‘Food_Consumption_exp1’ CSV file providing the mass of feeders at each 48hr time point. Continuous response variable. Variables Temp = temperature treatment (21[low]; 27[mid]; 30[high] Pesticide = control; imidacloprid; sulfoxaflor Time = the number of hours since onset of treatment exposure (0,48,96,144) Start - cumulative mass (grams) of feeders when first presented to the holding cages End - cumulative mass (grams) of the same feeders when being replaced Consumed - Start minus end
Experiment 2 files
Controlled Environment (CE) room conditions File names(s): ‘CE_Room_C_exp2’; ‘CE_Room_E_exp2’; ‘CE_Room_G_exp2’ CSV files providing humidity and temperature logger data evet minute (each row) over the period the experiment was run, showing high consistency. Continuous response variable. Humidity = Relative humidity in % Temp = Room temperarure in degrees Celsius
Feeding / consumption File names(s): ‘Food_Consumption_exp2’ CSV file providing the mass of sucrose solution consumed from feeders every 24hr time point. Continuous response variable. Variables Block = experimental replicate '1' or '2' Temperature = temperature treatment (21[low]; 27[mid]; 30[high] Pesticide = control; imidacloprid; sulfoxaflor Pot number = the ID for each holding cage per treatment Day - the day of the experiment Consumption - mass (grams) of sucrose solution consumed from the feeders per holding cage by the bees calculated by mass at start of provision minus the mass when replacing the feeders Bees alive = the number of bees still alive in each holding cage when replacing the feeders Fed per bee = mass consumed divided by the number of living bees
Movement and walking behaviour File names(s): ‘Master_results_NoStim_exp2’ CSV file providing data on dry body mass of tested bees and the suite of measured behaviours, including likelihood of movement and walking rates. Binary and continuous response variables. Variables Block = experimental replicate '1' or '2' Temperature = temperature treatment (21[low]; 27[mid]; 30[high] Pesticide = control; imidacloprid; sulfoxaflor Pot number = the ID for each holding cage per treatment Colony = parental colony the bees were taken from and placed into the holding cage Tag = the tag ID used to distinguish between workers in each holding pot Dry Mass = the dry mass of each bee in milligrams Time = number of hours since onset of treatment exposure. '0' represents baseline measure prior to treatment exposure State = active (A) or dead (D) Moved = yes (1) or no (0) Flew = yes (1) or no (0) Groomed = yes (1) or no (0) Fed = yes (1) or no (0) Time visible = amount of time (secs) during the video footage the bee was visible and not under the filter paper Visible = visible at any point during the recording - yes (1) or no (0) Time Flight = amount of time (secs) the bee was flying in the holding cage during the video footage Time Feed = amount of time (secs) the bee was feeding from a feederin the holding cage during the video footage Walk Time Available = amount of time (secs) remaining once the other behaviours had been accounted for that the bees had the potential to 'walk' Walked = yes (1) or no (0) Kinovea Walking Distance = the tracked distance (centimetres) that the bee walked Sdraw Walking Distance = corrected walking distance (see Methods in Kenna et al. 2023 GCB) Walking velocity overall = 'Sdraw Walking Distance' divided by the 'walk time available' Frames_moved = Number of video frames the bee walked during Time walked = total amount of time (secs) the beee actually walked Walking Velocity Active = 'Sdraw Walking Distance' divided by the 'time walked'
Walking video footage File names(s): ‘Worker5E_21S40_72hr_tracking video’ MKV file showing an example snap-shot recording of the workers in their holding cages with walking path tracked overlaid. The file name represents the worker ID (Worker5E), at the low temperature (_21), exposed to sulfoxaflor (S) at 40ug/L (40) and 72 hours after the onset of exposure (_72hr).
Experiment 3 files
Controlled Environment (CE) room conditions File names(s): ‘Room_C_Holding_exp3’; ‘Room_E_Flight_exp3’ CSV files providing humidity and temperature logger data evet minute (each row) over the period the experiment was run, showing high consistency. Continuous response variable. Variables Humidity = Relative humidity in % Temp = Room temperarure in degrees Celsius
Flight distance, duration, and velocity final measures File names(s): ‘Temp_Pest_Master_Spreadsheet_exp3’ CSV file providing flight performance data for each individual bee from the flight mill assay. Binary, count, and continuous response variables. Block = experimental replicate '1' or '2' Days since tag = the number of days the bee was tested since being tagged in the parental colony Start time = time of day the assay was run Bout = the bout the bee was flown in Temp = temperature treatment (21[low]; 27[mid]; 30[high] Food = control (C); imidacloprid (I); sulfoxaflor (S) Colony = parental colony the bees were taken from Bee weight nest = wet mass (grams) weight of the bee ITS = InterTegula Span (millimetres) Dry Mass = the dry mass of each bee in milligrams Fed = yes (1) or no (0) whilst in the holding tube and prior to the flight First feed = time (seconds) the bee fed during the first observed feeding event Second feed = time (seconds) the bee fed during the second observed feeding event Third feed = time (seconds) the bee fed during the third observed feeding event Fed Multiple = yes (1) or no (0) the bee performed multiple feeding events Feed Time = total time (seconds) feed for across first, second and third feeding events Slip weight = mass of the microscope cover slip (grams) Slip Droplet Weight Start = mass (grams) of the sucrose solution droplet when first provisoned Slip Droplet Weight End = mass (grams) of the sucrose solution droplet at end of being held in feeding tube Sucrose Consumed = 'Slip Droplet Weight Start' minus 'Slip Droplet Weight End' Relative Intake = 'Solution consumed' divided by 'ITS' (mg/mm) Consumption Rate = 'Sucrose Consumed' divided by 'Feed Time' (g/sec) Mass Specific Consumption = 'Sucrose Consumed' divided by 'Dry Mass' and multilied by 1000 to convert to milligrams (mg/mg) Bee Weight Fed = mass of bee after it fed Mill = the number ID of the flight mill used for that respective bee flight assay Initiated = yes (1) or no (0) the bee initiated flight on the fligh mill Bee Weight Flight = bee mass after the flight assay had ended Pre Clean Circuits = number of mill circuits the bee flew without data cleaning Strikes - Number of times the bee stopped flying during the 90 mninute test with 3 strikes terminating the trial Fly 0m = yes (1) or no (0) the bee flew more than zero metres after circuits had been cleaned Fly 90min = yes (1) or no (0) the bee flew for the full 90 mins before receiving three strikes Cleaned Circuits = number of mill circuits the bee flew once the data had been cleaned Distance = the distance (metres) the bee flew during the trial Duration = the duration (seconds) the bee flew during the trial Mean velocity = 'Distance' divided by the 'duration' Maximum velocity = the bout with the highest velocity calculated as the circumference of the rotated mill arm divided by the duration of that respective circuit
The following files were produced from the project, but due to the number of files and total size we have not uploaded to Dryad. However, we can provide these files on request by emailing the corresponding author Richard Gill (r.gill[at]imperial.ac.uk).
Experiment 2
The separate snap-shot videos of the holding cages that were taken for each cage every 24 hours over the experiment. Each video follows the format described above.
Walking path coordinate text files generated through the Kinovea software for each video, and downstream R script for analysis.
Matlab script used to convert the 25 FPS videos to 6.25 FPS.
Experiment 3
The flight mill raw data output files (.DAT), and associated sata cleaning R script.
Fight trial metric files using the cleaned data for each flight bout containing number of circuits flown, distance, duration, mean velocity, and maximum velocity.
Experiment 1 - Analysis of responsiveness
File name: 'Experiment_1_Analysis.R' (R file) Description: Full R script for all data analysis for Experiment 1
Experiment 2 - Analysis of movement, walking, and consumption behaviours.
File name: 'Experiment_2_Analysis.R' (R file) Description: Full R script for all data analysis for Experiment 2
File name: 'Track_Path_Plotting.R' (R file) Description: Script to calculate the distance walked by each bee from the Kinovea generated coordinate text file. There is a script with loop to load in all coordinate files, remove the frames the bee didn’t move, calculate a distance and walking speed (over the frames that the bee moved), and export a master data file containing this info for every worker that moved.
File name: 'Video_Frame_Rate_Script.m' (M File) Description: Matlab script used to convert the 25 frame per second tracking videos into 6.25 frame per second.
Experiment 3 - Analysis of flight performance
File name: 'Experiment_3_Analysis.R' (R File) Description: Full R script for all data analysis for Experiment 3
File name: 'Data_Cleaning.R' (R File) Description: Script used to clean raw data files and produce the flight trial metric files and the flight trial velocity files