When a pathogen infects a number of different hosts, the process of determining the relative importance of each host species to the persistence of the pathogen is often complex. Removal of a host species is a potential but rarely possible way of discovering the importance of that species to the dynamics of the disease. This study presents the results of a 12-year programme aimed at controlling brucellosis in cattle, sheep and goats and the cascading impacts on brucellosis in a sympatric population of red deer (Cervus elaphus) in the Boumort National Game Reserve (BNGR; NE Spain). From February 1998 to December 2009, local veterinary agencies tested over 36 180 individual blood samples from cattle, 296 482 from sheep and goats and 1047 from red deer in the study area. All seropositive livestock were removed annually. From 2006 to 2009 brucellosis was not detected in cattle and in 2009 only one of 97 red deer tested was found to be positive. The surveillance and removal of positive domestic animals coincided with a significant decrease in the prevalence of brucellosis in red deer. Our results suggest that red deer may not be able to maintain brucellosis in this region independently of cattle, sheep or goats, and that continued efforts to control disease in livestock may lead to the eventual eradication of brucellosis in red deer in the area.