Although Phlebotomus argentipes as the only known vector of visceral leishmaniasis (VL) is zoophilic in nature, VL is considered to be anthroponotic in the Indian subcontinent. Peripheral blood samples from 85 stray dogs were examined for any molecular evidence of Leishmania infection in VL endemic areas of Bangladesh. Parasite DNA was detected in a blood sample from 1 of 85 (1.2%) stray dogs using ITS1-PCR, and PCR sequencing of the rRNA-ITS and cytochrome b gene confirmed that the parasitic DNA was Leishmania donovani. The results support the assumption that dogs are a probable animal reservoir for the Leishmania parasite in Bangladesh. It will be important to investigate the possible epidemiological role of dogs in domestic foci of VL endemic areas in Bangladesh.