Outline of the test

1) The AI or formulation of a PPP is tested.

2) The PPP is dissolved in acetone if possible. Other solvents should be used only in instances where the compound is insolvable in acetone and these alternative solvents are known to be harmless to bees.

3) When formulations are tested (rather than AIs), they should be water and if needed, an appropriate wetting agent added. If a wetting agent is applied, it should be applied in the positive and negative reference as well.

4) The test substances are administered to anaesthetized bees (Human et al., 2013) in a 1µl droplet on the dorsal thorax of individual bees.

5) After treatment, the bees are provided ad libitum with freshly made sucrose-solution 50% (w/v) and checked daily for mortality and behaviour (see Tab 2).