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

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.

The seasonal dynamics and biting behavior of potential Anopheles vectors of Plasmodium knowlesi in Palawan, Philippines.

BACKGROUND: A small number of human cases of the zoonotic malaria Plasmodium knowlesi have been reported in Palawan Island, the Philippines. Identification of potential vector species and their bionomics is crucial for understanding human exposure risk in this setting. Here, we combined longitudinal surveillance with a trap-evaluation study to address knowledge gaps about the ecology and potential for zoonotic spillover of this macaque malaria in Palawan Island. METHODS: The abundance, diversity and biting behavior of human-biting Anopheles mosquitoes were assessed through monthly outdoor human landing catches (HLC) in three ecotypes representing different land use (forest edge, forest and agricultural area) across 8 months. Additionally, the host preference and biting activity of potential Anopheles vectors were assessed through comparison of their abundance and capture time in traps baited with humans (HLC, human-baited electrocuting net-HEN) or macaques (monkey-baited trap-MBT, monkey-baited electrocuting net-MEN). All female Anopheles mosquitoes were tested for the presence of Plasmodium parasites by PCR. RESULTS: Previously incriminated vectors Anopheles balabacensis and An. flavirostris accounted for > 95% of anophelines caught in longitudinal surveillance. However, human biting densities were relatively low (An. balabacensis: 0.34-1.20 per night, An. flavirostris: 0-2 bites per night). Biting densities of An. balabacensis were highest in the forest edge, while An. flavirostris was most abundant in the agricultural area. The abundance of An. balabacensis and An. flavirostris was significantly higher in HLC than in MBT. None of the 357 female Anopheles mosquitoes tested for Plasmodium infection were positive. CONCLUSIONS: The relatively low density and lack of malaria infection in Anopheles mosquitoes sampled here indicates that exposure to P. knowlesi in this setting is considerably lower than in neighboring countries (i.e. Malaysia), where it is now the primary cause of malaria in humans. Although anophelines had lower abundance in MBTs than in HLCs, An. balabacensis and An. flavirostris were caught by both methods, suggesting they could act as bridge vectors between humans and macaques. These species bite primarily outdoors during the early evening, confirming that insecticide-treated nets are unlikely to provide protection against P. knowlesi vectors.

INVESTIGATION OF MOSQUITOES WITH EMPHASIS ON AEDES (FINLAYA) POICILIUS, PUTATIVE VECTOR OF BANCROFTIAN FILARIASIS ON PANAY ISLAND, THE PHILIPPINES.

Entomological investigations were conducted in four remaining lymphatic filariasis endemic provinces of Panay Island, The Philippines to determine mosquito species present in these areas and to identify host preference and biting activity of Aedes (Finlaya) poicilius Theobald, primary vector of nocturnal periodic Wuchereria bancrofti in The Philippines. Sampling techniques targeted nocturnally active mosquito species using a carabao-baited trap (CBT) and human-landing collection (HLC), the latter taking place from 06:00 to 12:00 pm. A total of 25,536 mosquitoes comprising 42 species and 7 genera were collected from CBT, whilst HLC acquired 6,486 mosquitoes comprising 28 species and 5 genera. Three known or potential vectors of human filarial were collected, namely, Aedes poicilius, Culex quinquefasciatus and Mansonia uniformis. The peak landing (biting) activity for Ae. poicilius was between 09:00 and 11:00 pm. Comparisons between CBT and HLC yields showed this species to be more zoophilic. Based on observed mosquito behavior and interviews with residents, vector-host contact was promoted by the local practice of staying overnight in makeshift shelters in high risk areas without adequate protection against mosquito bites. Results of this survey will augment information for integrating vector control and mass drug administration into an island-wide lymphatic filariasis elimination program.

BIONOMICS AND ECOLOGY OF ANOPHELES LITORALIS ON BONGAO ISLAND, TAWI-TAWI PROVINCE, PHILIPPINES: IMPLICATIONS FOR VECTOR CONTROL.

Entomological surveys were conducted to identify Anopheles malaria vector species, their feeding and resting behaviors, and characterization of larval habitats on Bongao Island, Tawi-tawi Province, in July and November, 2007. Survey parameters included all-evening human-landing collections (HLC), evening buffalo-baited trap (BBT) collections, daytime indoor and outdoor adult resting collections, adult female age-grading, identification of natural Plasmodium infections in mosquitoes, larval habitat identification and physical/biological characterization, and adult insecticide susceptibility assays. Both surveys revealed the predominant and putative malaria vector species on Bongao Island is Anopheles litoralis. Anophelesflavirostris was collected on only one occasion. The HLC during the July survey produced approximately 4 mosquitoes/human/night (mhn). The November survey yielded 1.27 mhn due, in part, to inclement weather conditions during time of sampling. Anopheles litoralis host seeking behavior occurred throughout the evening (06:00 PM - 06:00 AM) with peak biting between 10:00 PM and 04:00 AM. This species exhibited stronger zoophilic behavior based on comparison of HLC and BBT data. HLC showed a slightly greater exophagic (outdoor) behavior (1.4:1 ratio). During the July collection, an older adult population was present (75% parous) compared to the lower numbers of An. litoralis dissected in November (25% parous). Albeit a small sample size (n=19), 10.5% of An. litoralis dissected contained midgut oocysts of Plasmodium. Daytime adult resting harborages included biotic and abiotic sites in and around partially shaded, brackish water habitats where immature stages were common. Anopheles litoralis was found susceptible to pirimiphos-methyl and four different synthetic pyrethroids. This survey provides further epidemiological evidence of the importance of An. litoralis in malaria transmission on Bongao Island, and presumably throughout much of the Sulu Archipelago in the southern Philippines. Published observations of this species remain very limited and further investigations on the bionomics and epidemiological importance of this species are needed. Both ecological and human factors in malaria transmission are presented with implications for improved control of An. litoralis and prevention of infection.