Adverse social and welfare implications of mixing dairy cows or separating calves from their mothers have been documented previously. Here we investigated the behavioral and physiological responses of individuals remaining after conspecifics were removed. We conducted a series of 4 experiments incorporating a range of types of different dairy cattle groupings [experiment 1 (E1), 126 outdoor lactating dairy cows; experiment 2 (E2), 120 housed lactating dairy cows; experiment 3 (E3), 18 housed dairy calves; and experiment 4 (E4), 22 housed dairy bulls] from which a subset of individuals were permanently removed (E1, n = 7; E2, n = 5; E3, n = 9; E4, n = 18). Associations between individuals were established using near-neighbor scores (based upon identities and distances between animals recorded before removal) in El, E2, and E3. Behavioral recordings were taken for 3 to 5 d, before and after removal on a sample of cattle in all 4 experiments (El, n = 20; E2, n = 20; E3, n = 9; E4, n = 4). In 2 experiments with relatively large groups of dairy cows, El and E2, the responses of cows that did and did not associate with the removed cows were compared. An increase in time that both nonassociates and associates spent eating was observed after conspecific removal in E1. In E2, this increase was restricted to cows that had not associated with the removed cows. A reduction in ruminating in remaining cattle was observed in E3 and eating in E4. Immunoglobulin A concentrations increased after separation in both E3 and E4 cattle, but did not differ significantly between associates and nonassociates in E2. Blood and milk cortisol concentrations were not affected by conspecific removal. These findings suggest that some animals had affected feeding behavior and IgA concentrations after removal of conspecifics.