DNA Barcoding of Potential Mosquito Disease Vectors (Diptera, Culicidae) in Jazan Region, Saudi Arabia

Abstract

The conventional morphological characterization of mosquito species remains heavily used for species identification in Jazan, Saudi Arabia. It requires substantial expertise and time, as well as having difficulty in confirming identity of morphologically similar species. Therefore, to establish a reliable and accurate identification system that can be applied to understanding spatial distribution of local mosquito species from the Jazan region, DNA barcoding was explored as an integrated tool for mosquito species identification. In this study, 44 adult mosquito specimens were analyzed, which contain 16 species belong to three genera of potential mosquito disease vectors (Aedes, Anopheles, and Culex). The specimens were collected from the Jazan region located in southwest Saudi Arabia. These included old and preserved mosquito voucher specimens. In addition, we assessed the genetic distance based on the generated mitochondrial partial COI DNA barcodes to detect cryptic diversity across these taxa. Nine mosquito species belonging to three genera were successfully barcoded and submitted to GenBank, namely: Aedes aegypti, Aedes caspius, Aedes vexans, Aedes vittatus, Anopheles arabiensis, Culex pipiens, Culex quinquefasciatus, Culex sitiens, and Culex tritaeniorhynchus. Of these nine species, Aedes vexans, Aedes vittatus, Culex sitiens, and Culex tritaeniorhynchus were registered in GenBank for the first time from Saudi Arabia. The DNA barcodes generated a 100% match to known barcodes of these mosquito species, that also matched with the morphological identification. Ae. vexans was found to be either a case of cryptic species (subspecies) or a new species from the region. However, more research has to be conducted to prove the latter. This study directly contributes to the development of a molecular reference library of mosquito species from the Jazan region and Saudi Arabia. The library is essential for confirmation of species in support of existing mosquito surveillance and control programmes.

Modification of the Folmer primers for the cytochrome c oxidase gene facilitates identification of mosquitoes

“…The FCOS universal primers have also been used for mosquito identification by many researchers [15,[21][22][23][24][25][26][27][28][29][30][31][32]. However, in our studies and those of others [5,8,29,[33][34][35], it has been noted that FCOS sometimes fails to adequately identify mosquito species. We thus re-examined the FCOS and developed primers better suited for mosquito identification.…”

“…Our finding that not all mosquito DNA could be amplified with AUCOS has been reported previously with FCOS. The age of the specimens prior to extraction has been implicated, with museum and voucher specimens proving particularly difficult to amplify [ 29 , 35 ]. This might have played a role in our study, as both Saint Kitts and Auburn have warm tropical climates that might lead to rapid desiccation, death, and DNA damage in mosquitoes trapped early in the 24-h trapping periods we used.…”

“…This might have played a role in our study, as both Saint Kitts and Auburn have warm tropical climates that might lead to rapid desiccation, death, and DNA damage in mosquitoes trapped early in the 24-h trapping periods we used. It is of note that Noureldin et al [ 35 ] used damp cotton pads in their traps and had very high success rates for amplifications with their PCRs using shortened and modified Folmer primers. Differences in DNA extraction methods were suggested to account for FCOS success rates varying from 43 to 85% by other workers [ 5 ].…”

Toxicity of plants as insecticides against human pathogenic mosquito vectors of Saudi Arabian strains—A review

“…Among them, mosquitoes (Diptera: Culicidae) speaks to be a key threat for millions of organisms worldwide. Culicidae family relays larger species of 3300 with 41 genera (Kalaivani et al 2012), later researcher reveals 112 genera and 3547 species (Harbach 2018, Noureldin et al 2022. They act as a vectors for significant parasites and pathogens causing malaria, dengue, filariasis, West Nile, Zika virus (Benelli 2015;Murugan et al 2017), tick-borne Lyme disease and rickettsioses (Paules et al 2018), chikungunya (Puntasecca et al 2021), yellow fever (Bicca-Marques et al 2022), Japanese encephalitis (Bhatt & Sharma 2014), lymphatic filariasis, Rift Valley fever, St. Louis encephalitis (Mayer et al 2017;Benelli et al 2018).…”

“…Culicidae family relays larger species of 3300 with 41 genera (Kalaivani et al . 2012), later researcher reveals 112 genera and 3547 species (Harbach 2018, Noureldin et al 2022). They act as a vectors for significant parasites and pathogens causing malaria, dengue, filariasis, West Nile, Zika virus (Benelli 2015; Murugan et al .…”