Variation of cacophony and period gene regions in sand fly species (diptera: Psychodidae) in Turkey

Sand flies are important in transporting many pathogens which are closely related to human and animal health. Specimen identification is the most important step in establishing the vector-pathogen relationship correctly. While specimen identification of sand flies basically relies on traditional approaches, it is difficult to make a robust discrimination between the members of species complexes and between closely related species using only morphological characters. Genes related to sand fly behavior have been reported to be successful in distinguishing between the closely related species and species complexes. The aim of this study was to reveal the variations within and between the sand fly species collected from a wide range of geographical origins in Turkey using cacophony and period genes that are related to courtship behavior of sand flies. The analyses were conducted using a total of 884 specimens belonging to 16 species/species complexes that are the representatives of Turkish sand fly fauna. Intraspecific and interspecific distances were estimated, phylogenetic trees were constructed and genetic analyses of populations were performed after the amplification of above mentioned genes. The discrimination power of these genes between the different species and members of species complexes were also discussed. Results obtained for cacophony gene showed that the interspecific distance values range between 1.34% - 14.36% in Paraphlebotomus subgenus, 4.23% - 10.34% in Larroussius subgenus, 0.93% - 31.86% in Adlerius subgenus, 7.55% - 5.15% in Transphlebotomus subgenus and 18.00% in Sergentomyia subgenus. For the period gene, the interspesific distance values were ranged between 1.94% -12.77% in Paraphlebotomus subgenus and estimated as 6% and 7% in Larroussius and in Adlerius subgenera, respectively. Both cacophony and period genes evaluated in this study were found to be successful in discrimination of all sand fly species in Turkey, and helpful for the delineation of the members of the species complexes. Moreover, using the cacophony gene a possible hybridization event was identified between the members of Phlebotomus major complex for the first time. Evaluation of all results together showed that both cacophony and period genes are important molecular markers which can be used in sand fly taxonomy.