In Ecuador, as in other Latin American countries, bovine tuberculosis (BTB) is a major concern in the dairy cattle industry, with economic and public health importance. The increase of peri-urban dairy production associated with the absence of BTB control program in Ecuador are major risk factors in the context of tuberculosis. Furthermore, official data related to the disease are scarce and there is no national veterinary reference laboratory for mycobacteria in Ecuador. The present work was undertaken in this context. The main objectives were to provide data on the epidemiologic situation of BTB among dairy cattle in northern Ecuador, and to compare the performances of the standard diagnostic tools used to identify M. bovis. The data presented confirm the importance of BTB in dairy cattle in northern Ecuador and highlight the need for a national BTB control program. Ideally, this program should include (1) a reference laboratory for BTB and the training of qualified veterinarians; (2) mandatory skin testing and culling of reactors; (3) detailed post-mortem inspection with laboratory-based diagnosis with a trace-back system and (4) information campaigns in rural areas.