The World Health Organization considers leishmaniasis to be one of the most important zoonotic diseases with approximately 350 million people in 98 countries at risk of contracting the disease. The leishmaniasis also remains a severe public health problem in Turkey. It is important to evaluate the prevalence of Canine leishmaniasis (CanL) for designing control policies. In the present study, we report for the first time the occurrence of Leishmania infection in dogs living in the Middle Black Sea Region of Turkey. For the diagnosis of the infection parasitological (direct microscopic examination of lymph nodes), serological (ELISA) and molecular (kDNA-PCR) methods were performed using blood samples obtained from 240 dogs. Sand flies were also collected using CDC miniature light traps from the same locations for testing Leishmania presence by kDNA-PCR. Only one (0.41%) dog out of 240 was found to be positive by both tests. Amastigotes of Leishmania spp. were detected in the same animal lymph node aspirate sample. A total of 18 sand flies were collected and no Leishmania was detected Phlebotomus spp. sample pool. Our results showed that CanL could have public health implications and future screening studies are needed in this region of Turkey.