Various Morphologic Shapes of Leishmania major Amastigotes with low Genetic Structure among People Afflicted with Zoonotic Cutaneous Leishmaniasis in Khuzestan Province

Tegumentary leishmaniasis has been implicated in many types of different phenotypes due to diverse genetic extent with polymorphic amastigote shapes. Molecular diversity and morphological changes of Leishmania major (L. major) have turned into a controversial issue among researchers. This investigation revealed the features of amastigote polymorphic shapes in clinical manifestations of suspected patients to zoonotic cutaneous leishmaniasis (ZCL) in the direction of genetic variation in Khuzestan province.
Of 315 suspected patients, sampling, smear preparation and staining were carried out during 2012-2014. After DNA extraction, the PCR products were digested with BsuRI endonuclease enzyme in the process of RFLP by targeting ITS-rDNA, Cyt b and Microsatellite genes.
L. major and L. tropica were the parasites identified in patients having regular amastigote's shapes (oval or round) with a size of 2-4 μm in each of dry, wet and classical mixed lesions. Whereas, irregular shapes (spindle, pear or cigarette) were observed in non-classical wet lesions with more than 4 μm. Sequencing of ITS-rDNA and Cyt b genes of L. major demonstrated low molecular variation (P >0.05) except for one common haplotype (GenBank access no. EF413075)
On the basis of present findings, there is no significant correlation between phenotypic and genotypic features of L. major isolates. This study is considered as the first exhaustive report to incriminate morphometric shapes of L. major amastigotes concerning their relevance with various clinical appearances and genotypic characteristics.