DOI: 10.5061/dryad.j6q573nr3
title: Data for "Visual Threat Avoidance While Host Seeking by Aedes Aegypti Mosquitoes"

description: The mosquito Aedes aegypti infects hundreds of millions of people annually
 with disease-causing viruses. When a mosquito approaches a host, the host
 often swats defensively. Here, we reveal the mosquito’s escape behavior
 during host seeking in response to a threatening visual cue – a newly
 appearing shadow. Reactions to a shadow were far more aversive when it
 appeared quickly, versus slowly. Remarkably, mosquitoes evaded shadows
 under very dim light conditions. We knocked out the gene encoding the TRP
 channel, which compromised the ability to avoid threatening shadows, but
 only under relatively high light conditions. Conversely, removing two of
 the five rhodopsins expressed in the compound eyes, Op1 and Op2,
 diminished shadow aversion, but only under low light. Following the
 removal of a threatening visual cue, mosquitoes quickly re-initiate host
 seeking. Thus, female Aedes balance their need to host seek with visual
 threat avoidance by rapidly transitioning between these two behavioral
 states.
description: To cast a moving shadow on the mosquitoes, we placed a 22-cm-long
 belt-driven moveable stage 33 cm in front of the behavioral cage. Attached
 to the stage was a custom-fabricated mount that secured a 13.5 cm x 6 cm x
 1.9 mm clear plexiglass window. On top of this window, we placed a
 rectangular strip of white, opaque electrical tape (3 cm x 6 cm), which
 when moved in front of our white LED light source (Figure S1C; Outerbanks
 Provisions) blocked visible light (Figures 1B and 1C). Movement of the
 stage was achieved by a Nema-17 stepper motor, which was wired to an A4988
 motor driver (StepperOnline), set to 4V using the builtin potentiometer.
 The motor driver received movement commands from an Arduino Uno (Egloo),
 and the speed of the stage was modified by changing the pause length
 between each motor step. The mosquitoes were recorded with a varifocal
 webcam (ELP), affixed with an IR-pass filter (Heliopan), at 30 frames per
 second, and at a resolution of 640 x 480 pixels. To provide constant
 illumination for the experiments, we placed near IR (850 nm) LED lights
 (Waveform) behind our behavioral cage. The IR lights were shown through a
 translucent white acrylic panel (30.5 cm x 30.5 cm x 3 mm), which served
 as a light diffuser.
description: # Data for "Visual threat avoidance while host seeking by Aedes
 aegypti mosquitoes"
 [https://doi.org/10.5061/dryad.j6q573nr3](https://doi.org/10.5061/dryad.j6q573nr3) ## Description of the data and file structure These are the data for **"Visual threat avoidance while host seeking by *Aedes aegypti* mosquitoes".** Each figure has a corresponding Excel file containing information on which videos were used and the raw data from each video. The Excel file labeled 'BehaviorVideoList' has a list of the videos used for each figure panel with the video name represented as a date such as 031124. These videos are then broken down into zip files by date so that the video would be within the 030124-032924 zip. Each dated folder contains a video along with the files generated by our code to analyze the videos.  Below is the data explanation of each source and extended data Excel file. All time data is in seconds - SourceData_Figure2: ID refers to each track with time being the first column. Each track in a column has values relevant to the figure panel (Figure 2E sheet has the percentage of landed mosquitoes on the back wall over time).  - SourceData_Figure3: The raw data is contained in `MarkovChainData_noShadow.mat`.  - SourceData_Figure4: Tab 4A-4D - Spontaneous takeoffs is a percentage of mosquitoes taking off. Tab 4E - Video is the name of the video used, the OnBackWall column is the whole number of mosquitoes on the back wall, Spontaneous takeoffs is the percentage of mosquitoes taking off randomly, Raw takeoffs is a percentage of mosquitoes taking off in response to the shadow. Condition is related to what type of experimental condition was used for that video. Cage is which cage A-F was used. Replicate is a number 1-3 related to the first second or third time the cage was run. Genotype is the type of mosquito used, Normalized takeoffs is the percentage takeoff rate of raw takeoffs minus the spontaneous take offs. Tab 4F - speed is in cm/s, the best fit line was made using the values underneath that column. Tab 4H- ExperimentID is the name of video folder (which is a date), UpAvgTO is the orientation of UP mosquito's average take off percentage rate, DownAvgTO is the orientation of down mosquito's average take off percentage, UpNumberMosquito is the whole number of mosquitos in the up orientation, DownNumberMosquito is the whole number of mosquitoes in the down orientation, AvgSpontTO is the average spontaneous take off percentage rate, NormalizedTakeoffsUp is the normalized take off rate in the up orientation, NormalizedTakeoffsDown is the normalized take off rate in the down orientation T-test is the output of the t-test on the data. Tab 4I follows the same naming scheme as 4H but in left or right instead of up and down. Tab 4J - The data is the normalized take off rate percentage, the columns are the light levels in lux and the rows are the speed of the shadow in cm/s. - SourceData_Figure5: 5A - The data within this tab is all the percentage of landed mosquitoes (who many mosquitoes out of 30 are on the back wall). The first column is time in seconds, with increasing time. Each column is a condition followed by the replicate number. Male human* *odor1 is the male mosquitoes presented with human odor cues on the back wall and the first replicate. Male sucrose 1 is male mosquitoes presented with sucrose on the back wall and the first replicate. Female unstarved 1 is female mosquitoes fed on sucrose presented with host cues on the back wall and the first replicate. 5B - The data within this tab is the percentage of searching mosquitoes (the amount of mosquitoes walking around on the back wall searching). The first column is time. The following columns follow the same naming scheme as 5A, with the addition of female_starved, which are female mosquitoes starved of sucrose for 24 hours then presented host seeking cues on the back wall. male starved are male mosquitoes starved of sucrose then presented sucrose. 5C - The video is the name of the video, OnBackWall is the whole number of mosquitoes landed on the back wall, Spontaneous takeoffs is the percentage of mosquitoes taking off randomly, Raw takeoffs is a percentage of mosquitoes taking off in response to the shadow, Lux is the light level condition in lux, Cage is the cage A-F that was used for the biological replicate, sex is male or female mosquito, Fed is either starved of sucrose or fed on sucrose, replicate is number 1-3 related to first second or third time the cage was run. normResp_mean is the mean value of the takeoffs, the condition is related to the sex and feeding status like male starved.  - SourceData_Figure6: 6B - genotype is wildtype or trp transheterozygous, relative trp expression is the relative expression amount and bio rep is the biological replicate. 6D-6O - The first column is time and then each next column is the mosquito number. The data values within each column related to the mv position on the erg plot. 6P-6Q - age is the day age of the mosquito day 1, 5 or 10. genotype is trp transheterozygote or wildtype liverpool. Lux is the light level in lux, trace is the mosquito number, stimNumber is 1 meaning it is related to the first erg light stimulus, final_amplitude is the erg amplitude in response to the stimulus.  - SourceData_Figure7: 7B - video is the video name, OnBackWall is the whole number of mosquitoes landed on the backwall, Spontaneous Takeoffs is the percentage of takeoffs randomly happening, RawPropTO is the raw takeoff percentage in response to the shadow, lux is the light level in lux, genotype is the genotype 9trp transheterozygote or liverpool wildtype, cage is A-F cage of mosquitoes, off edge is the first or second off edge shadow pass, replicate is the first second or third replicate of that cage, normalized takeoffs is the raw takeoffs minus the spontaneous takeoffs. 7D - the naming scheme is the same but the shadow number column is related to the first, second, third or fourth shadow pass. 7E - the naming scheme is the same there are just more genotypes tested here included in the genotype column (op1, op2 and op1op2). - ExtendedData_Figure1: Wavelength is in nm or nanometers, relative_int is the relative intensity value of our light source. - ExtendedData_Figure2: S2A - vidID is the video name, lux is the light level in lux and totalTrackLength is the length of time in seconds that a mosquito was on the back wall. S2B - avg searching bout length is the average amount of time in seconds that a mosquito searches on the backwall. S2C - avg stationary bout length is the average amount of time in seconds that a mosquito sits on the back wall without taking off or starting searching. S2D the raw data is in compiled TOTable.mat and these are the time steps. S2E - video is the name of the video, light is the Light to dark transition (LD) or dark to light to transition (DL), StageMoveFrame is the frame number when the stage is moving, OnOnBackWall is the whole number of mosquitoes on the back wall, BSProp TO mean is the mean percentage of take offs spontaneously happening, RawPropTO is the percentage of takeoffs happening in response to the shadow, Date is the date of the video, cage is the cage A-F labeled, Tech_rep is the replicate on the cage (first second or third), normalized takeoffs is the rawPropTO minus the spontaneous takeoffs. S2H - has a similar naming scheme and freezing is the percentage of mosquitoes responding to the shadow by freezing movement. Off-Edge shadow speed is in cm/s of the speed of the shadow. - ExtendedData_Figure3: A: First column is time in seconds, the rest of the data is percentage of mosquitoes searching. The two column conditions are females responding to host cues or females responding to sucrose. B: Cage is the cage label A-F, condition is host seeking cues on the back wall or sucrose on the back wall, mean p searching is the percentage of mosquitoes searching. C: video is video name, replicate is the replicate on the cage, cage is the label on the cage A-F, sex is female or male mosquitoes, light is LD (light to dark shadow) or DL (dark to light shadow), OnBackWall is the whole number of mosquitoes on the back wall, BSPropTO mean is the percentage of spontaneous take offs averaged over the condition, RawPropTO is the raw percentage of takeoffs in response to the shadow, Fed is whether or not the mosquitoes were starved of sucrose, normResp mean is the normalized percentage of take offs averaged over the condition, condition is the host seeking cues or sucrose on the back wall.  - ExtendedData_Figure5: S4A-S4D - Time is in seconds, voltage is the erg value in mv, genotype is control for wildtype liverpool line or trp transheterozygotes. S4E - file name is the name of the file, block the number of shadows passed, genotype is wildtype liverpool line or trp transheterozygotes, dark time is the amount of time they are in the dark in seconds, base line is the first value in the erg in response to no stimulus, depol is how far the voltage moves from the stimulus (depolarization), amplitude is the change in mv from baseline to depol.  - ExtendedData_Figure6: S6A - genotype is wildtype liverpool line or trp transheterozygotes, light is the light level in lux, cage is the cage label A-F, mean on wall is the mean number of mosquitoes landed on the back wall. S6B - similar naming scheme to S6A, meanNormResp is the percentage of take offs normalized to the spontaneous take off rate. S6C - S6D are the same scheme.  Acronyms: TO = Take Off The code used for collecting raw data from each video is linked below in our GitHub repository.  ## Code/software [https://github.com/Craig-Montell-Lab/Meyerhof-et-al.-2025/tree/main](https://github.com/Craig-Montell-Lab/Meyerhof-et-al.-2025/tree/main)