Molecular Identification and Phylogenetic Analysis of Fleas Ctenocephalides Canis and Pulex Irritans in West and Northwest of Iran Based on Cytochrome Oxidase I

The fleas of the Pulicidae family are one of the most important external parasites of humans and domestic animals in the world. In addition to eating blood, these insects are carriers of some pathogens such as Yersinia pestis, Rickettsia typhi and Bartonella hensle to humans and animals. The aim of this study was to determine and evaluate the fauna of Ketonecefalides canis and Polex Irritants in western and northwestern Iran based on mitochondrial marker of cytochrome oxidase I.

The present study was a descriptive cross-sectional and phonetic study conducted in a period of 13 months from May 2018 to June 2019 in five provinces of Kermanshah, Kurdistan, Azerbaijan Western, Lorestan and Hamedan considering the prevalence of 10% and 95% confidence level with 5% error rate. In the present study, samples were collected by optical trap, human prey and direct isolation from the host. The samples were kept in 70% alcohol and identified in the parasitology laboratory using valid diagnostic keys. After DNA extraction and a Polymerase chain reaction (PCR), 20 samples were sent to Takapouzist Company for sequencing. The sequences were sequenced and compared using Emboos Needle and Omega software.

The collected samples included 918(47/39%) C.canis and 1019 (52/60%) P.irritant. The results of morphological studies revealed that there is no intraspecific difference between two species of fleas isolated from different hosts (p <0.05). However, the intraspecific difference based on the molecular marker of cytochrome oxidase I for ten studied populations was 0.15% in P.irritans and zero in C.canis. The difference between two genus of C. canis and P. irritans based on the molecular marker of cytochrome oxidase I was 14%.

No significant difference was observed in morphological characteristics of samples collected from different hosts. However, there were slight differences based on mitochondrial markers in the study populations. The results from the phylogenetic tree based on mitochondrial markers showed that despite the slight differences in this sequence of different hosts and cities, all samples from different regions are in the same phylogeny. The results of mitochondrial genome analysis indicated that these pieces are useful for demonstrating intraspecific similarity, and differentiation at species level and genus.