Genetic characteristics and phylogenetic analysis of Leishmania infecting sandflies and humans based on the kDNA and ITS2 gene dataset in the new focus of Cutaneous Leishmaniasis in Shiraz, south of Iran

Abstract

Abstract Objective Cutaneous leishmaniasis is a neglected tropical disease with an incidence of one million people annually worldwide. Iran has many important fuci for this disease which Leishmania major and L. tropica cause. However, this study aimed to investigate Zoonotic and environmental factors in the spread of leishmaniasis in the city of Shiraz, southern Iran with a phylogenetic inference of Leishmania by examining the infection of patients and vectors with Leishmania parasite based on molecular datasets of kDNA and ITS2 genes. Methods Sand flies were caught using sticky paper traps indoors (bedrooms, bathrooms, storage rooms, etc.) and outdoors. Twenty samples of human lesions were also examined. Leishmania infection was evaluated by PCR of kinetoplast (k) DNA. Subsequently, parasite species identification was confirmed by PCR and DNA sequencing of the 18S ribosomal RNA internal transcribed spacer two (ITS2) gene. Results All Phylogenetic reconstruction based on Bayesian, UPGMA, and Neighbor-Joining methods revealed sequences of Leishmania parasites infecting patients and sand-fly species were clustered into two main monophyletic clades of L. major and L. tropica. The overall infection rate of sand flies (pools) with Leishmania DNA was 72.5%, and the Mean diversity within L. major clade was 0.01. Conclusions A new focus on ACL and ZCL is reported in the tourist city of Shiraz. Human interference in the environment, such as habitat destruction, housing in orchards and foothills, and other health factors, have caused the emergence of both diseases in this area which need to be considered for a survival health program.