12.6.1. Example 1: Nosema infection for life-long monitoring

To study the effects of Nosema infections,

1. Mark 300 -500 newly emerged bees at the thorax (see section 2.3.).

2. Introduce the young bees into a nucleus bee hive which serves as the test hive.

3. After four days, collect 140 of the colour-marked bees from the combs of the registration hive.

4. Divide the bees in the following way:

4.1. Label 60 bees with RFID-tags.

4.2. Divide these bees into two subgroups; each group of 30 bees is confined to a cage.

5. Bulk-feed both groups with honey solution (33 % weight/volume). The control group (Nosema-) receives pure sucrose solution, while the Nosema group (Nosema+) receives honey solution contaminated with Nosema spores (5 x 106 spores/millilitre, for experimental infestation, see the BEEBOOK paper on Nosema; Fries et al., 2013). 6. The remaining 80 bees are needed for subsequent determination of a successful Nosema infection. These bees are only required for infection control and therefore are not RFID-tagged. Instead, label these bees with an additional, different colour-marking: e.g. blue for Nosema- and red for Nosema+ group.

7. Distribute these bees for infection control evenly over the cages of the RFID-tagged bees, so that each cage now contains 70 bees.

8. When all of the bees in each cage have consumed the honey solution offered completely, re-introduce them into the registration colony with a delay of 15-30 minutes between the cages to avoid trophallaxis between inoculated and non-inoculated bees.

9. Twelve days later, collect the colour-marked positive and negative control bees from the hive and examine them for Nosema infection (see the BEEBOOK paper on Nosema; Fries et al., 2013).

10. Leave the RFID-labelled bees in the colony and do not disturb them again. Their foraging behaviour can be studied throughout their entire live as described above.