184.108.40.206.2. Experimental setup
1. Clean a small glass Petri dish (60 mm diameter) with acetone and hexane or pentane.
2. Place 2 dead adult bees at 2 diametrically opposite sides of the Petri dish, close to the walls (Fig. 10).
3. Treat one bee with the substance tested, treat the other (control) bee with the solvent used to transfer the tested substance on the first bee.
Use a volume of solvent as small as possible to avoid perturbing the layer of cuticular hydrocarbons. In case of a removal / restoration bioassay the bees' cuticle need be washed with a solvent to remove the hydrocarbons before the tested profile is applied.
4. Place the Petri dishes in a thermostatic cabinet, in darkness, at 34.5 °C and 70 % RH.
5. Place one adult female mite in the centre of the Petri dish.
6. Note mite position every 10 min for 60 min.
Three positions are considered: mite on the treated bee, mite on the control bee, mite not on bees.
7. Test 10 mites in different Petri dishes simultaneously and replicate 6 times.
8. Alternate side of treated and control bees for each replicate to control for the influence of external factors on mite locomotion.
Fig. 10. Test arena for phoretic mite attraction cues.