D3482: The mighty mitogenome: Advancing barcoding strategies for identifying cryptic mosquito species

Mosquito species diversity can have a significant impact on vector-borne disease incidence. Many mosquito species that have been implicated in driving disease transmission, belong to cryptic species complexes. Accurate identification of these medically important vectors is necessary for successful mitigation strategies, with current molecular tools proving to be problematic for resolving members belonging to species complexes. Sequencing strategies have revealed that the full mitochondrial genome (mitogenome) is useful in rectifying species classification in comparison to traditionally reference molecular barcodes using fragments of the cytochrome oxidase unit I (COI) and the ribosomal DNA internal transcribed spacer 2 (ITS2). Here we describe a genome skimming strategy to capture mitogenomes for rectifying mosquito species identification and taxonomic classification. The main objective of this study was to develop a new source of reference genetic barcodes for multiple mosquito species. Single mosquito specimens were sequenced with raw reads obtained via Illumina Miseq V2 chemistry and Oxford Nanopore technologies MinION platforms. Full mitogenomes were generated for Anopheles gibbinsi, Aedes albopictus, Ae. aegypti, Culex quinquefasciatus and Haemagogus janthinomys. This study demonstrates that skimming for the mitogenome is a relatively inexpensive and efficient approach for species identification and concurrent taxonomic characterization for multiple mosquito species, as well as generating reference sequence data for the community to build upon.