A total of 565 mongrel dogs from rural localities of Venezuela were examined by serological (DAT, IFAT and ELISA) and parasitological tests to address the status of Trypanosoma cruzi infection and to evaluate their role in the transmission of the infection to human population. The overall percentage of sero-positive infected dogs shown to be 67.6% (382/565):253 (61.7%) from 47 villages belonging to 8 states located at 4 different geographical regions of western Venezuela and 129 (33.5%) dogs from 48 households located in areas where Chagas disease is endemic. From 101 sampled dogs living in close proximity to 30 acute chagasic patients, 84% expressed specific anti-T. cruzi antibodies (Ab) with 12 of them (14%) showing blood circulating parasites (BCP). In these houses a high proportion of sero-positive people (20%) and frequent indoor infestation by triatomine-bugs (70%) was also recorded. The analysis revealed that from the 47 rural villages sampled during the study, 91.5% had the presence of T. cruzi sero-positive dogs, ranging from 62% positive localities at the states of Falcon and Cojedes to 100% in the other six studied Venezuelan states. This demonstrates that T. cruzi-infected dogs are found throughout all the geographical regions of western Venezuela irrespective of their ecological differences. Molecular typing of T. cruzi isolates from infected dogs using ribosomal and mini-exon gene markers, revealed the presence of both T. cruzi I and T. cruzi II lineages. The coincidence in the circulation of T. cruzi II in dog and human populations at the same locality and at the same time is reported and its significance is discussed. The combined serological, parasitological, epidemiological and molecular data is gathered here to call the attention on the presence of infected dogs as a risk factor in the maintenance of T. cruzi as a source for infection to humans.