We examined the sequence, order or steps of hygienic behavior (HB) from pin-killed pupae until the removal of them by the bees. We conducted our study with four colonies of Apis mellifera carnica in Germany and made four repetitions. The pin-killing method was used for evaluation of the HB of bees. The data were collected every 2 h after perforation, totaling 13 observations. Additionally, for one hygienic colony and another non-hygienic colony, individual analyses of each dead pupa were made at every observation, including all details, steps or sequences of HB.
Most research on hygienic behavior has recorded the time taken by the colony to remove an experimental amount of dead brood, usually after one or two days. We evaluated the time that hygienic (H) and non-hygienic (NH) honey bees take to uncap and remove dead brood in observation hives after the brood was killed using the pin-killing assay. Four experimental colonies were selected as the extreme cases among 108 original colonies. Thirty brood cells were perforated with a pin in two H and two NH colonies and observations were made after 1, 2, 3, 4, 5, 6, and 24 h.