Isolation process of two genes of wsp and 16S rRNA in the intracellular bacterium; Wolbachia pipientis in Phlebotomus papatasi sandfly vector of Zoonotic Cutaneous leishmaniasis in Iran

The intracellular Wolbachia pipientis is a Rickettsia-like bacterium. The gene of W. pipientis is usually congenitally-inherited in insects and can cause cytoplasmic incompatibility (CI),which is potentially useful for driving genes through populations.In present study, W. pipientis has been detected in Phlebotomus papatasi, the vector of rural cutaneous leishmaniosis in Iran by using PCR to amplify fragments either of the 16S ribosomal RNA gene (16S rRNA) or of the major Wolbachia surface protein gene wsp. sandflies were screened for the presence of W. pipientis by PCR using the nonstrain specific primers wsp 81F with 691R and 16S 99F with 994R. The sequences obtained were edited and aligned using SequencherTM v. 3.1 to identify W. pipientis haplotypes, which were analysed phylogenetically using PAUP* software. from 165 Individual wild-caught P. papatasi from Iran, the gene fragment of wsp was found in 124 cases (75.2%), and 16s RNA gene fragment was identified in 123 cases (74.5%). Only one haplotype was obtained for each gene, from which it is inferred that only one A-group genetic strain of W. pipientis occurs in P. papatasi throughout much of this sandfly's range. based on the results of this study there is possibility of using just one genetically modified strain of W. pipientis to drive through wild sandfly populations transgenes for intervening in the transmission of L. major by P. papatasi. This genetically modified strain would have to show a CI phenotype in P.papatasi, in order to be spread quickly through wild sandfly populations. Due to natural infection of Wolbachia in sandflies and for their behavior of Cytoplasmic Incompatibility(CI), Wolbachia can be used as a transferring gene in populations of sandflies to control Leishmaniasis.