A part of data used in the analysis represented in Figure 2. We studied behavioral change associated with orphaning. Each raw represents each specific individual. The same individual appears both before (control) and after orphaning. Data of Colony M1 are presented. We used a scanning technique (Altmann 1974) to observe the behavior of all individually marked adults (n = 35 to 77) in each of five colonies. The interval between scans was at least 10 min, and each individual was scanned at least 100 times during 4 days to allow estimation of the time budget for each behavioral act. The gamergates were then removed and the scanning was repeated for another 4 days. We compared the frequencies of resting, selfish behavior, and social behavior before and after removal of the gamergates. The selfish behavior consisted of dominance behavior and self-grooming. The social behavior included brood care, allo-grooming (donating), food handling, nest construction, and outside-nest activities. We regarded walking within the nest as non-specific behavior, because its function was difficult to identify. We treated motionless ants as resting, even when they were being groomed by other workers. We examined the difference in the frequency of each of the four major behavioral categories (resting, selfish behavior, social behavior, and the other non-specific behavior) between before and after orphaning. The change in the relative frequency of each behavioral category was examined by the Wilcoxon matched pair signed rank test using individual workers as blocks for each colony. Then we combined the results of colonies using unweighted Stoufferfs z-score method (Whitlock 2005). We repeated these analyses focusing on dominant and subordinate workers, which were distinguished by observation under the orphaned condition, as defined previously.