Multi-level selection for hygienic behaviour in honeybees.

Disease is one of the main factors driving both natural and artificial selection. It is a particularly important and increasing threat to the managed honeybee colonies, which are vital in crop pollination. Artificial selection for disease-resistant honeybee genotypes has previously only been carried out at the colony-level, that is, by using queens or males reared from colonies that show resistance. However, honeybee queens mate with many males and so each colony consists of multiple patrilines that will vary in heritable traits, such as disease resistance. Here, we investigate whether response to artificial selection for a key resistance mechanism, hygienic behaviour, can be improved using multi-level selection, that is, by selecting not only among colonies as normal but also among patrilines within colonies. Highly hygienic colonies were identified (between-colony selection), and the specific patrilines within them responsible for most hygienic behaviour were determined using observation hives. Queens reared from these hygienic patrilines (within-colony selection) were identified using DNA microsatellite analysis of a wing-tip tissue sample and then mated to drones from a third highly hygienic colony. The resulting colonies headed by queens from hygienic patrilines showed approximately double the level of hygienic behaviour of colonies headed by sister queens from non-hygienic patrilines. The results show that multi-level selection can significantly improve the success of honeybee breeding programs.