title: Different Morphologies of Leishmania major Amastigotes with No Molecular Diversity in a Neglected Endemic Area of Zoonotic Cutaneous Leishmaniasis in Iran.
abstract: Molecular diversity of Leishmania major and its morphological changes have become a controversial issue among researchers. Some aspects of polymorphic shapes of amastigotes in clinical manifestations along with molecular variation were evaluated among suspected patients of some exceptional zoonotic cutaneous leishmaniasis locations in Northern Khuzestan, Southwestern Iran.
  
  
  
  Suspected patients (n = 165) were sampled in zoonotic cutaneous leishmaniasis foci over two consecutive years during 2012-2014. Prepared smears were stained, scaled and measured by ocular micrometer. DNA was extracted from smears; ITS-rDNA and Cytochrome b (Cyt b) markers were amplified, and PCR products were digested by BsuR1 restriction enzyme. Then the RFLP and sequencing were employed.
  
  
  
  Only L. major was identified in patients containing regular amastigotes' shapes (oval or round) with a size of 2-4 µm in each of classical wet, dry, mixed lesions. Meanwhile, irregular shapes (spindle, pear, or cigarette) were observed separately in non-classical wet lesions with more than 4 µm. Interestingly, a few amastigotes with an external flagellum were observed in some lesions. All sequenced ITS-rDNA and Cyt b genes of L. major did not show any molecular variation (χ 2 P > 0.05), including only one common haplotype (GenBank access no. EF413075).
  
  
  
  Findings proved that unlike other endemic foci, there is not a meaningful correlation between phenotypic and genotypic features of L. major isolates. This study is considered as the first comprehensive report to incriminate morphometric shapes of L. major amastigotes, which enhances our knowledge concerning their relevance with various clinical appearances and genotypic traits.
authors.affiliation: Molecular Systematics Laboratory, Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran.|Dept. of Parasitology, Medical Faculty, Shahid Beheshti University of Medical Sciences, Tehran, Iran.|Dept. of Parasitology, Medical Faculty, Tabriz University of Medical Sciences, Tabriz, Iran.
authors.forename: Adel
authors.initials: A
authors.lastname: Spotin
authors.affiliation: Dept. of Parasitology, Medical Faculty, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Soheila
authors.initials: S
authors.lastname: Rouhani
authors.affiliation: Molecular Systematics Laboratory, Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran.|Dept. of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.
authors.forename: Parnazsadat
authors.initials: P
authors.lastname: Ghaemmaghami
authors.affiliation: Dept. of Parasitology, Medical Faculty, Shahid Beheshti University of Medical Sciences, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Haghighi
authors.affiliation: Dept. of Microbiology, Qom Branch, Islamic Azad University, Qom, Iran.
authors.forename: Mohammad Reza
authors.initials: MR
authors.lastname: Zolfaghari
authors.affiliation: 5Dept. of Epidemiology, Medical 
  Sciences of Tehran branch, Islamic Azad University, Tehran, Iran.
authors.forename: Aref
authors.initials: A
authors.lastname: Amirkhani
authors.affiliation: Molecular Systematics Laboratory, Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Mahin
authors.initials: M
authors.lastname: Farahmand
authors.affiliation: Molecular Systematics Laboratory, Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Ali
authors.initials: A
authors.lastname: Bordbar
authors.affiliation: Molecular Systematics Laboratory, Dept. of Parasitology, Pasteur Institute of Iran, Tehran, Iran.
authors.forename: Parviz
authors.initials: P
authors.lastname: Parvizi
mesh_terms: D000818:Animals; D019353:Endemic Diseases; D006801:Humans; D007492:Iran / Q000453:epidemiology; D018320:Leishmania major / Q000235:genetics*; D016773:Leishmaniasis, Cutaneous / Q000453:epidemiology* / Q000469:parasitology
keywords: Amastigote shapes; Iran; Leishmania major; Mitochondrial gene; Nuclear gene