Tehran University of Medical Sciences Journal of Arthropod-Borne Diseases 2322-1984 9 1 2015 10 11 84 97 EN Alireza Zahraei-Ramazani Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Dinesh Kumar Department of Zoology, Centre of Advanced Study, Banaras Hindu University, Varanasi 221 005, India. Hossein Mirhendi Department of Medical Parasitology and Mycology, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Shyam Sundar Department of Medicine, Institute of Medical Sciences, Banaras Hindu University, Varanasi 221 005, India. Rajnikan Mishra Department of Zoology, Centre of Advanced Study, Banaras Hindu University, Varanasi 221 005, India. Vahideh Moin-Vaziri Department of Parasitology and Mycology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. Hassan Soleimani Yazd Health Research Station, National Institute of Health Research, Yazd, Iran. Mohammad Reza Shirzadi Communicable Disease Management Centre, Ministry of Health and Medical Education, Tehran, Iran. Reza Jafari Esfahan Health Research Station, National Institute of Health Research, Esfahan, Iran. Ahmad Ali Hanafi-Bojd Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. Sodabe Hamedi-Shahraky School of Health, Zabol University of Medical Sciences, Zabol, Iran. Mohammad Reza Yaghoobi-Ershadi Department of Medical Entomology and Vector Control, School of Public Health, Tehran University of Medical Sciences, Tehran, Iran. 2015 10 10 2015 10 10 Background: Female sand flies of subgenus Adlerius are considered as probable vectors of visceral leishmaniasis in Iran. The objective of this study was to determine the morphological and genotypic variations in the populations of this subgenus in the country. Methods: Sand flies collected using sticky traps from 17 provinces during 2008–2010. The morphometric measurements were conducted with an Ocular Micrometer. Data was analyzed by SPSS. The Cytb gene was used to estimate population genetic diversity and identify the female specimens. UPGMA phenetic tree was used for DNA haplotypes of Cytb gene. Results: Six species of subgenus Adlerius identified from which one species, P. (Adlerius) kabulensis, is new record. The identification key is provided for males. Results revealed the molecular systematic in the species of subgenus Adlerius and determine the relationship of three females of P. comatus, P. balcanicus and P. halepensis. Conclusion: The positions of three females and the males in the UPGMA tree are correct and the similarities among them confirm our results. The branches of each species are not genetically distinct which justify the overlapping morphological characters among them. Molecular sequencing of Cytb-mtDNA haplotypes can be used for female identification for different species of subgenus Adlerius in Iran. https://jad.tums.ac.ir/index.php/jad/article/view/120 https://jad.tums.ac.ir/index.php/jad/article/download/120/108