Intensively housed dairy cattle are commonly regrouped (mixed into a new social group) as part of routine farm procedures. This stressful procedure triggers heightened levels of agonistic behaviors and disrupts animals' time budgets. However, little is known regarding the effects of regrouping on cattle's affective states. The aim of this study was to explore whether regrouping (involving a change in both the social and physical environment) triggers anhedonia (i.e., the reduced ability to experience pleasure) in 6-mo-old dairy heifers, a phenomenon associated with negative mood. In this study, we assessed anhedonia using changes in the use of a mechanical brush. Holstein heifers (n = 16) were trained to use a mechanical brush and then given the opportunity to individually brush for 10 min every 2 d. Time spent brushing (during a 10-min brush test) was collected before, during, and after regrouping (2-d interval) with the assumption that heifers would reduce their use of the brush during regrouping. Each heifer was individually regrouped into a new social group composed of 12 older and unfamiliar heifers and allowed access to the brush at 8 and 56 h after the onset of regrouping. Immediately after the last test, each heifer was brought back to her original pen and allowed to mingle with familiar pen-mates before being tested again 2 and 4 d later. When tested 8 h after regrouping, heifers reduced time spent brushing by 44 ± 27% (95% confidence interval: −96.18 to −41.8) compared with before regrouping; however, no differences were detected 56 h after regrouping. There was no relationship between the intensity of the decrease in brush use and any behaviors (number of agonistic interactions received, time heifers spent resting, or synchronization at the feed bunk) recorded for the 8 h immediately before testing (i.e., between 0 and 8 h and between 48 and 56 h after regrouping). These results indicate that regrouping induces anhedonia-like responses in dairy heifers on the day of regrouping. This routine procedure may thus induce negative mood in dairy heifers. This response was not related to behaviors typically collected to assess the negative effects of regrouping. Maintaining dairy cattle in stable social groups should be favored.