Datesets corresponding to the below methods: Landing_tent_Ae. aegypti, Landing_tent_Ae.notoscriptus, Landing_tent_Ae.vigilax

Data was collected as follows:

We conducted experiments on mosquito attraction to humans using three species: Ae. aegypti, Ae. notoscriptus and Ae. vigilax. The experiments were conducted in a 3 Å~ 3 m tent under constant light levels and at room temperature. Each trial involved releasing 100 males, aged between one and two weeks, that had previously been allowed to mate, into the tent. The males were given 30 min to acclimatize before the experiment began. The experiments were filmed using GoPro Hero 10 cameras placed at either end of the tent, with white panels (84.1 Å~ 118.9 cm) as a background. In each trial, one side was baited with a human subject, while the other side was left unbaited as a control. Subjects stood facing the camera with their bare feet and shins in the field of view, with this position remaining consistent across trials. Subjects did not wear any perfume. The side of the baited and unbaited treatment was alternated for each trial. The number of trials, human subjects and number of days of the experiments are summarized in electronic supplementary material, table S1. The same batch of males was used for multiple trials on the same day but replaced daily. Treatments were recorded for 30 min using the time-lapse function immediately after the human subject assumed their position inside the tent. The number of mosquitoes in view of the camera was scored every 20 s, distinguishing between males that were in flight and males that landed on the human subject. For data analysis, we calculated the average number of male mosquitoes in each category over the entire trial period.

Interpretation of the datasets:

Columns:

• Species: the species tested
• Person: which human subject was tested at the time
• Treatment: either human or control. Control here means blank; human means subject was tested
• Observation: 1- 90 counts (every second frame was counted)
• Landed: number of mosquitoes that were landed on the human subject at the given observation
• Flying: number of mosquitoes that were flying in camera view at the given observation
• Total: the number of landed and flying mosquitoes combined at the given observation

Datesets corresponding to the below methods: Female_preferences_Ae.aegypti, Male_preferences_Ae.aegypti, Female_preferences_Ae.notoscriptus

Mosquito preferences for different human subjects

In our experiments, we found that Ae. aegypti males exhibit preferences towards certain human subjects (see electronic supplementary material, figure S3). While previous research has demonstrated differential attraction of female Ae. aegypti mosquitoes to different human hosts [16,37,38], this has not yet been quantitatively reported in males. We conducted additional experiments in which we used a consistent set of five human subjects (coded A–E) who stood in pairs in opposite positions in the tent set-up described in §2.3.2. The subjects were filmed for 5 min on each side before the sides were swapped and the procedure was repeated. This was done for each pairwise combination of the five subjects (20 combinations in total), with a fresh batch of males being used for each day of four separate days. The footage was scored as described in §2.3.2. For data analysis, we calculated the average number of male mosquitoes in view (combining flight and landed) over the 5 min of each trial for each human subject. We then tested all pairwise combinations of the same five human subjects for their attraction to female Ae. aegypti and Ae. notoscriptus. We used a two-port olfactometer (30 Å~ 30 Å~ 30 cm) similar to those used in previous studies by Ross et al. [39] and Amos et al. [34]. The mosquitoes used in this experiment were 6–7 days post-emergence and had been allowed to mate prior to the experiment. We released approximately 50 females into the set-up and allowed them to acclimatize for approximately 1 min. A box fan placed at the opposite end of the cage drew air through two traps into the cage. Pairs of subjects placed one hand each in front of one of the traps. After 5 min, we closed the entrance to the traps and counted the number of females in each trap and individuals remaining in the cage. The combinations of subjects and sides were randomized until all 20 pairwise comparisons between subjects and sides were completed. We repeated the experiment using the same five subjects for another 4 days using a fresh batch of females each day for a total of 10 replicates (five per side). Aedes vigilax females were not assessed in this experiment due to relatively low rates of attraction to humans observed in a pilot trial using this olfactometer design.

Prior to data analysis, we calculated a preference index for each person to reflect the relative attraction of each subject by dividing the number of mosquitoes attracted to one human subject by the number of mosquitoes attracted to both subjects. We determined the average preference index for all replicates of each human subject. Statistical analyses were performed using the preference index averaged across replicates.

Effect of mosquito repellent on male mosquitoes that show attraction to humans After we confirmed that male Ae. aegypti show attraction to humans in our tent experiments, we tested whether they are repelled by a commercial mosquito repellent (Aerogard tropical strength insect repellent, Reckitt Benckiser, NSW, Australia) containing 191 g kg−1 diethyltoluamide and 40 g kg−1 N-octyl bicycloheptene dicarboximide. We used the same tent set-up as described in §2.3.2. The repellent was applied to the knees downwards to one of the two human subjects positioned on either site of the tent within 5 min before the trial began. The number of males in view was recorded every 20 s for 10 min. The person wearing the repellent and the sides of the treatment and control were randomized. We ran 20 trials over 5 days with a rotation of nine human subjects, with the batch of 100 Ae. aegypti males replaced each day. The footage was scored as described in §2.3.2. For data analysis, we calculated the average number of male mosquitoes in view (combining flight and landed) over the 5 min of each trial.

Interpretation of the columns of the tables:

• Species: which species was tested
• Left: human subject that was tested on the left hand site of the tent
• Right: human subject that was tested on the right hand site of the tent
• Observation: 1- 15 counts (every second frame was counted)
• Landed: number of mosquitoes that were landed on the human subject at the given observation
• Flying: number of mosquitoes that were flying in camera view at the given observation
• Total: the number of landed and flying mosquitoes combined at the given observation
• Percent: the precent of attracted mosqutioes for the human subject compared to the human subject they were tested against. This was then used to calculate a preference index for each subject to test whether certain human subjects were overall more attractive than others.

If you have any questions about the data presented here, please email me at: veronique_paris@hotmail.de
I am very happy to provide more detail or explain any analysis.