Morphological and Molecular Identification of Aedes Aegypti through Genetic Marker Mitochondrial Encoded Cytochrome C oxidase I (MT-COI) Gene in Pakistan

Abstract

Aedes aegypti are slender, fragile, long-legged mosquito, which are distributed worldwide. In general, it is a terrestrial and nocturnal species that act as pollinator for few plant species as well as it is a hematophagous insect and act as main vector for many arboviruses (dengue, yellow fever, etc.). In many cities of Pakistan, dengue fever is becoming more prevalent as an infectious disease. as there is currently no vaccine for the dengue virus and control of the disease depends on vector control. The present study aims to investigate morphological and molecular identification of Aedes aegypti in district Buner. A total of 365 samples of mosquitoes were randomly collected from six tehsils of the district Buner. Before examination, all the collected samples were preserved in labelled Eppendorf tubes containing silica gel and transported to Veterinary Research & Disease Investigation Center (VR&DIC), Balogram Swat, Khyber Pakhtunkhwa, Pakistan for further laboratory analysis. Morphological identification was performed with the help of stereomicroscope using taxonomical keys. To increase identification accuracy, each sample was reconfirmed by PCR using mitochondrial Cytochrome oxidase first gene (COI). The amplified PCR product was sequenced, and the obtained sequences were trimmed by Bio Edit and BLAST in NCBI.  After BLAST, phylogenetic tree was constructed by using MEGA-X. In phylogenetic tree, the sequence (COI) genes of Aedes aegypti species as previously reported from Sri Lanka and India. Furthermore, the results of this study will generate basic information about mosquitos spp; and it will help to understand the epidemiology of Aedes aegypti disease in Khyber Pakhtunkhwa, especially in district Buner, Khyber Pakhtunkhwa, Pakistan.