3.6.2.5.2. Experimental setup

This bioassay is described by Ziegelmann et al. (2012).

1. Queen cell cups (e.g. Nicot system®) can be used as test arena.

It is recommended to embed the cell cups in a glass Petri dish with wax.

2. Ensure a temperature of 28-30 °C in the cell cup by placing the setup on a hotplate.

3. Transfer the relevant mite stages into the cell cup.

When extracts or single substances are tested, follow steps 4 and 6, if only behaviour is observed, go to 6.

4. Apply volatile test substances to a piece of filter paper (size: 1.5 mm x 15 mm).

5. Place volatile substances in the vicinity of the female; apply non-volatile substances directly to the female mite.

For the application of test substance, chose a solvent which does not harm or repel the female.

6. Cover with a glass plate to prevent mites from escaping.

7. Record the male responses with e.g. the Observer software (Noldus Information Technology) for 5 or 10 minutes.

8. Categorise male responses as follows: 1. movement around female; 2. mounting the female’s dorsum; and 3. copulation attempt on the female’s venter.