Two seroepidemiological surveys on canine leishmaniosis in stray dogs were performed annually in the Madrid region for 10 years (November 1996-April 2006). The presence of anti-Leishmania antibodies was detected by immunofluorescence antibodies test (cut off 1:100). The overall seroprevalence found in the 1,803 dogs studied was 7.8% (141 positive dogs). Seropositivity was not associated with either breed or sex. Statistical analysis revealed greater seroprevalence in groups of older dogs, indicating that the probability of exposure to the bite of sand flies infected with Leishmania infantum increased with age. The most important result was a high proportion of seropositivity for leishmaniosis (79.5%) among dogs without clinical signs of canine leishmaniosis. These data are very important because stray dogs can play an important role in the epidemiology of this zoonotic disease. Furthermore, the stray population could be useful sentinels to follow the progress of the disease in endemic areas.