['To test the hypothesis that Mycobacterium bovis can persist in the environment within protozoa.', 'In this study, we used a novel approach to detect internalized mycobacteria in environmental protozoa from badger latrines. Acid-fast micro-organisms were visualized in isolated amoebae, although we were unable to identify them to species level as no mycobacteria were grown from these samples nor was M. bovis detected by IS6110 PCR. Co-incubation of Acanthamoeba castellanii with virulent M. bovis substantially reduced levels of bacilli, indicating that the amoebae have a negative effect on the persistence of M. bovis.'] Do environmental amoebae support the long-term survival of virulent mycobacteria?