5.2.2.2.5.4.1. Condition of the colonies

1) The condition of the colonies is assessed once before the application and several times after the application according to the following time schedule:

   - BFD (brood area fixing day), first assessment
   - Application at +2 days (±1 day) after BFD
   - + 5 days (±1 day) after BFD
   - + 10 days (±1 day) after BFD
   - + 16 days (±1 day) after BFD
   - + 22 days (±1 day) after BFD
   -+ 28 days (±1 day) after BFD.

2) For the condition of the colonies the following parameters are assessed in order to record effects of the test chemical:

   - Colony strength (number of bees per Delaplane et al., 2013b)
   - Presence of a healthy queen (e.g. presence of eggs)
   - Pollen storage area and area with nectar or honey (per Delaplane et al., 2013b)
   - Area containing cells with eggs, larvae and capped cells (per Delaplane et al., 2013b).

The coverage of a comb can be estimated assuming that a comb is covered by 120 bees per 100 cm² if bees are sitting very close to each other (Imdorf and Gerig, 1999; Imdorf et al., 1987). The estimations will be done for all combs (both sides) in each hive. The assessment of the areas containing brood and food can be done by estimating subareas of 100 cm². Afterwards the number of cells per brood stage/food stock is calculated assuming that 100 cm² of the comb comprise 400 cells (Imdorf and Gerig, 1999; Imdorf et al., 1987). These estimations will be done for all combs (both sides) in each hive.

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