Genetic Determination of Phlebotomus Papatasi, Main Vector of Leishmania Major, Using Genetic Markers and Variety of DNA Haplotypes in Different Regions of Iran

The phlebotomine sandfly, Phlebotomus papatasi, is the main vector of Leishmania major, the causative agent of rural zoonotic cutaneous leishmaniasis in Iran. Knowledge of the genetic and population differentiation of sandflies will be useful for planning interventions in leishmaniasis transmission. Cytochrome b gene, Elongation factor–1 a gene and a surface protein gene of Wolbachia bacteria were used as markers. Sandflies were sampled from different regions of Iran using sticky papers, funnel traps, CDC miniature light traps and aspirator. Each sandfly was dissected in a drop of autoclaved 1 x TE on a clean microscope slide, to pull off the head and posterior abdomen to identify of sandflies and rest of body for extraction of DNA. Both primers were used for sequencing each sandfly DNA. Three fragments of Cytochrome b were amplified and sequenced. Thirty three CB1 haplotypes among the 189 sequences, Seven CB3 haplotypes among the 44 sequences and 49 Cyt b Long haplotypes among the 149 sequences were identified. The prevalence of the surface protein gene of Wolbachia was high. Elongation factor-1 a gene implified by PCR. However, some part of fragment often gave sequences that were not readable. Phylogenetics analysis of all the three fragments of Cyt b identified a single haplotype network. Unlike Cyt b, the wsp gene of Wolbachia did not show any polymorphism which are consistent with the absence of cryptic sibling species of P. papatasi. No well-differentiated lineage was found, and no lineage predominated in different habitats of Gerbil burrows and peridomestic animal shelters or geographical isolation by distance.