Background: Trypanosoma cruzi is a parasite protozoa that infects mammalians and in the human cause Chagas' disease, which represent a major health problem in Colombia where an estimated of 436,000 individuals are infected, with 11% of the population at risk for contracting the disease. Moreover, the potential epidemiological significance of bats as possible reservoir hosts for T. cruzi, has been previously remarked. Different neotropical bats species have been reported to be susceptible to T. cruzi infection. They participate in important ecological processes and because of its ability to fly can spread infectious diseases from the natural environment to the homes of people. In Colombia, few studies on bats in endemic areas for Chagas' disease have been performed. Thus, we evaluated the presence of T. cruzi in heart tissue taken from bats Cordoba department (northern Colombia), considered an endemic area for this infection. Methods & Materials: 30 hearts of bats were collected in four rural localities from Cordoba department. The DNA was purified using a commercial high-purity PCR template preparation kit (Roche, Mannheim, Germany). The integrity of the purified DNA was analyzed through PCR amplification of the bat cyt b gene. PCR tests based on the TcH2AF-R and S35-S36 primers which amplify a fragment of SIRE element and a conserved region of minicircles from T. cruzi respectively, were evaluated for the detection of parasite in batś heart tissue Results: A total of 11 samples (36.6%) of three localities were positives for both PCR. Three species were positive for the presence of T. cruzi: Carolia perspicillata and Dermanura phaeotis (frugivorous) and Molossus molossus (insectivore). Conclusion: This is the first report of T. cruzi in the heart of naturally infected bats in Colombia. These findings imply that there is an active transmission of parasite among bats populations from Cordoba. Therefore, it is important to continue assessing how bats natural infection can be acquire and spread the parasite, since these species are highly distributed in the region and human intervention in their natural ecosystems is contributing the migration to urban areas, which increase parasite circulation in the disease domestic transmission cycle