Tehran University of Medical Sciences Journal of Arthropod-Borne Diseases 2322-1984 18 3 2024 09 30 253 263 Narges Marvi Moghadam Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mehdi Mohebali Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Yavar Rassi Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Ali Reza Zahraei-Ramazani Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mohammad Ali Oshaghi Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Reza Jafari Isfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mahboubeh Fatemi Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Mohammad Hossein Arandian Isfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Hamid Abdoli Isfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Nilufar Shareghi Isfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Maryam Ghanei Isfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Nilufar Jalali-Zand Isfahan Health Research Station, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Arshad Veysi Zoonoses Research Center, Research Institute for Health Development, Kurdistan University of Medical Sciences, Sanandaj, Iran Javad Ramazanpoor Isfahan Health Center, Isfahan University of Medical Sciences, Isfahan, Iran Kurosh Aminian Isfahan Health Center, Isfahan University of Medical Sciences, Isfahan, Iran Ali Salehi Isfahan Health Center, Isfahan University of Medical Sciences, Isfahan, Iran Ali Khamesipour Center for Research and Training in Skin Diseases and Leprosy, Tehran University of Medical Sciences, Tehran, Iran Amir Ahmad Akhavan Department of Vector Biology and Control of Diseases, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2023 06 22 2023 07 16 Background: Zoonotic cutaneous leishmaniasis (ZCL) is an endemic disease in Varzaneh City where Leishmania ma­jor is the causative agent and the great gerbil, Rhombomys opimus, is the main reservoir host of the disease. Despite control efforts, ZCL outbreaks recur every few years. This study was conducted to revive information on the parasite/s species circulating between humans and the reservoirs in the region. Methods: Leishmania infection in patients and R. opimus was studied using direct parasitological and molecular methods during 2019–2021. Nested-PCR and DNA sequencing were used for Leishmania parasite identification. Inter and intra-species variations in the Leishmania parasites were investigated using BLAST and MEGA7 software. Results: All suspected patients (N=34) and 14 out of 36 great gerbils tested positive for Leishmania parasites via direct parasitological method. Nested-PCR method revealed all the patients were infected with L. major (94.1%) and mixed infection of L. major and Leishmania turanica (5.9%), and great gerbil specimens were infected with either L. major (44.4%), L. turanica (5.6%), or Leishmania gerbilli (5.6%) and also with mixed infection of L. major and L. turanica (30.5%), L. major and L. gerbilli (8.3%) and mix of all the three Leishmania species (5.6%).   Conclusion: The identical sequences of L. major in both human patients and rodents indicate that the great gerbils are the main reservoirs of L. major in Varzaneh City. The presence of L. turanica in patients would be of interest to carry out further studies to determine the role of this species in the persistence, signs, and treatment of ZCL in humans. https://jad.tums.ac.ir/index.php/jad/article/view/1656 https://jad.tums.ac.ir/index.php/jad/article/download/1656/668