Tehran University of Medical Sciences Journal of Arthropod-Borne Diseases 2322-1984 16 4 2023 02 27 1465 1465 Azad Absavaran Department of Medical Entomology & Vector Control, 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 Vahideh Moin-Vaziri Department of Medical Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran Alireza Zahraei-Ramazani Department of Medical Entomology & Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Amir Ahmad Akhavan Department of Medical Entomology & Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran Sayena Rafizadeh Ministry of Health and Medical Education, Tehran, Iran Amirhossein Rassi Faulty of Medicine, Tehran University of Medical Sciences, Tehran, Iran Alireza Barmaki - Yavar Rassi Department of Medical Entomology & Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran 2021 07 06 2023 02 22 Background: The primary aim of this study is to determine infection to Leishmania parasites in the wild population of Phlebotomus caucasicus and Phlebotomus mongolensis using molecular methods in some important zoonotic cutaneous leishmaniasis foci in Iran. Methods: Sand flies were collected from active colonies of rodent burrows from 16 trapping sites using sticky trap pa­per. In order to detect and identify of Leishmania parasites in females Ph. caucasicus and Ph. mongolensis, the Nested–PCR amplification of ITS2-rDNA region was performed to generate amplicon with 245bp for Leishmania major, 206bp for L. gerbilli and 141bp for L. turanica. Results: In the current study we found DNA of different gerbil parasites such as L. major and L. turanica, and mixed infection of L. major/L. turanica in Ph. caucasicus and Ph. mongolensis. It should be noted that, in Iran, natural infec­tion with Leishmania parasites is recorded for the first time in this study in Ph. mongolensis. Conclusion: Both species of Ph. caucasicus and Ph. mongolensis not only may participate in the ZCL transmission cycle between reservoir hosts, but also results of this study support the  role of these species as secondary vectors in the transmission of leishmaniasis to humans. https://jad.tums.ac.ir/index.php/jad/article/view/1465 https://jad.tums.ac.ir/index.php/jad/article/download/1465/609