Control of malaria-transmitting mosquitoes using gene drives

Gene drives are selfish genetic elements that can be re-designed to invade a population and they hold tremendous potential for the control of mosquitoes that transmit disease. Much progress has been made recently in demonstrating proof of principle for gene drives able to suppress populations of malarial mosquitoes, or to make them refractory to the Plasmodium parasites they transmit. This has been achieved using CRISPR-based gene drives. In this article, I will discuss the relative merits of this type of gene drive, as well as barriers to its technical development and to its deployment in the field as malaria control.This article is part of the theme issue ‘Novel control strategies for mosquito-borne diseases''.     

The effect of permethrin-impregnated bednets on a population of Anopheles farauti in coastal Papua New Guinea

The effect of introducing bednets impregnated with 0.4 g/m2 permethrin on local populations of the malaria vector mosquitoes Anopheles farauti Laveran and An. koliensis Owen was monitored in a coastal village of Papua New Guinea. Whole-night landing collections were undertaken for 25 consecutive nights before and 21 nights after the introduction of the nets. Capture-recapture experiments and resting collections were also performed before and after the introduction of the nets. Following the introduction of treated nets, the biting population of An. farauti, the predominant vector, dropped from an average of 689 to 483 per man-night and the oviposition cycle became irregular, although survival rates (determined by time series analysis of the landing catches and log regression of recapture rates) were not significantly affected. The densities of An. farauti resting in and around houses and the human blood index of the engorged females also decreased significantly after introduction of the treated bednets. The population of An. koliensis dropped prior to the introduction of the nets. However, the number of nulliparous females in the landing catches remained more or less constant which implies that, in this species, survival rates were affected by the nets but that recruitment to the population was not.     

Effects of physico-chemical treatments on hemagglutination activity of Anopheles gambiae haemolymph and midgut extract

Abstract. Anopheles gambiae midgut extracts and haemolymph possessed agglutinins, titre 1:16 to 1:256, against human red blood cells (RBCs). Subjection of both tissues to protein precipitation reagents, organic chemical and selected protease, neuraminidase and other glycosidic hydrolase treatments revealed the haemagglutinins to be protein, most likely glycoprotein, in nature - not lipoprotein, lipid, glycolipid or nucleic acid. An.gambiae agglutinins were thermo-labile >40^oC, affected by freezing and thawing treatments, and contained disulphide and hydrogen bonds on the basis of sensitivity following exposure to dithio-threitol and urea respectively. Optimum haemagglutination depended generally on slightly acid to neutral pH conditions and agglutinin activity was Ca²⁺ ion, albeit to a lesser extent Mg²⁺ ion, dependent. The midgut extract agglutinin subunit molecule had a relative molecular weight (M_r) of 65kDa whilst that of haemolymph was 40kDa.
This study presents the first report on selected physico-chemical properties, the glycoproteinaceous nature and tentative subunit M_r of mosquito midgut extract and haemolymph anti-RBC agglutinin(s).     

Physiological changes governing the onset of sexual receptivity in male mosquitoes

The effectors which erect antennal hairs in male mosquitoes are shown to be unresponsive to the neurotransmitter that triggers antennal hair erection until 12–24 hr after emergence.The unresponsive period can be prolonged by transfusion of haemolymph from newly-emerged males, or by injection of 20-hydroxyecdysone. This suggests that the hormone persisting after the metamorphosis from pupa to adult, affects the duration of the post-emergence delay in antennal hair erection and is indirectly responsible for timing the onset of sexual receptivity in male mosquitoes.     

Malaria in macaques

Seven species of malaria naturally infect eight species of macaques in southwestern India, Sri Lanka, and Southeast Asia. Within malarious areas, the frequency of infections in infected species of macaques varies from 9 to 61%. Natural malarial infections in macaques are relatively benign. The proved or probable vectors of macaque malaria are seven species of mosquitoes that belong to the Leucosphyrus Group of the genus Anopheles. The geographic distribution of macaque malaria apparently is determined by the distribution of the Leucosphyrus Group of mosquitoes, which in turn apparently is determined by the distribution of tropical evergreen rain forest. Experimental infections with three species of macaque malaria frequently are lethal to populations or species of macaques that inhabit areas outside the geographic ranges of the parasites. In populations or species of macaques that are sympatric with experimentally virulent species of malaria, partial resistance probably evolved as a consequence of natural selection acting on favorable mutations.     

Suppression of Aedes aegypti by predatory Toxorhynchites moctezuma in an island habitat

Larval populations of the mosquito Aedes aegypti were suppressed by predatory Toxorhynchites moctezuma mosquito larvae released systematically in a village on Union Island (Saint Vincent and the Grenadines) during March-December 1988. Eggs and larvae of Tx.moctezuma were transported from Trinidad and introduced into all semi-permanent and permanent water-holding containers in the experimental village at Clifton. The semi-isolated village of Ashton served as control. Base-line Ae.aegypti indices (house, ovitrap, Breteau, cistern/tank, drum/barrel, small containers) were obtained for the two villages over a 4-month period prior to the introduction of the predatory Tx.moctezuma mosquito larvae. After sustained releases of predators for 5 months, all indices of Ae.aegypti were lower in the treated village than in the untreated village during the last 3 months of the year.     

Close encounters: contributions of carbon dioxide and human skin odour to finding and landing on a host in Aedes aegypti

In a wind‐tunnel study, the upwind flight and source location of female Aedes aegypti to plumes of carbon dioxide (CO₂) gas and odour from human feet is tested. Both odour sources are presented singly and in combination. Flight upwind along the plumes is evident for both CO₂ and odour from human feet when the odours are presented alone. Similarly, both odour sources are located by more than 70% of mosquitoes in less than 3 min. When both CO₂ and odour from human feet are presented simultaneously in two different choice tests (with plumes superimposed or with plumes separated), there is no evidence that females orientate along the plume of CO₂ and only a few mosquitoes locate its source. Rather, the foot odour plume is navigated and the source of foot odour is located by over 80% of female Ae. aegypti. When a female is presented a plume of CO₂ within a broad plume of human foot odour of relatively low concentration, the source of CO₂ is not located; instead, flight is upwind in the diffuse plume of foot odour. Although upwind flight by Ae. aegypti at long range is presumably induced by CO₂ and the threshold of response to skin odours is lowered, our findings suggest that, once females have arrived near a prospective human host, upwind orientation and landing are largely governed by the suite of human odours, whereas orientation is no longer influenced by CO₂.     

Functional characterization of PGRP‐LC1 of Anopheles gambiae through deletion and RNA interference

Abstract Peptidoglycan recognition proteins (PGRP) play an important role in innate immunity in insects through the activation of the Imd pathway, which has been shown to be required in the antibacterial response in insects and in the limitation of the number of Plasmodium berghei oocysts developing in mosquito midgut. The LC1 gene of the PRGP family in Anopheles gambiae produces many products through alternative splicing. In this work, we demonstrate that PGRP‐LC1a alone is sufficient to activate the Imd pathway in the A. gambiae L3–5 cell line through a combination of terminal or internal deletions, and RNA interference against endogenous PGRP‐LC products. In the absence of endogenous PGRP‐LC proteins, the integrity of the cytoplasmic domain is necessary for LC1a function, while that of the extracellular domain is not. Moreover, the shorter the extracellular domain, the higher the activity for LC1a. However, the removal of either the cytoplasmic or the extracellular PGRP‐binding domain has little impact on the activity of LC1a in the presence of endogenous PGRP‐LC proteins.     

Wolbachia infection does not alter attraction of the mosquito Aedes (Stegomyia) aegypti to human odours

The insect endosymbiont Wolbachia pipientis (Rickettsiales: Rickettsiaceae) is undergoing field trials around the world to determine if it can reduce transmission of dengue virus from the mosquito Stegomyia aegypti to humans. Two different Wolbachia strains have been released to date. The primary effect of the wMel strain is pathogen protection whereby infection with the symbiont limits replication of dengue virus inside the mosquito. A second strain, wMelPop, induces pathogen protection, reduces the adult mosquito lifespan and decreases blood feeding success in mosquitoes after 15 days of age. Here we test whether Wolbachia infection affects mosquito attraction to host odours in adults aged 5 and 15 days. We found no evidence of reduced odour attraction of mosquitoes, even for those infected with the more virulent wMelPop. This bodes well for fitness and competitiveness in the field given that the mosquitoes must find hosts to reproduce for the biocontrol method to succeed.     

Avian defensive behavior and blood-feeding success of the West Nile vector mosquito, Culex pipiens

Avian defensive behavior against host-seeking arthropods influencestransmission of vector-borne pathogens by affecting mosquitobiting rate, either by preventing vector–host contactor by increasing the rate of multiple host feeding. We exposedhouse sparrows (Passer domesticus L.) and chickens (Gallus gallusdomesticus L.) to Culex pipiens pipiens L. overnight in a largeobservation cage and measured avian defensive behavior ratesand mosquito blood-feeding success. Both bird species exhibiteda range of defensive behaviors, 90% of which were foot stomps,head movements, and wing shakes. Total behavior rates increasedproportionately with mosquito density in both species, increasedafter the first hour of mosquito exposure, and decreased asindividual birds were exposed to mosquitoes multiple times.Mosquito blood-feeding success on house sparrows was high overall(82 ± 5%) and independent of behavior rates. Blood-feedingsuccess on chicks was lower (58 ± 5%) and negativelycorrelated with defensive behavior rate after the first hourof mosquito exposure. Results revealed a higher percentage ofpartial blood meals on chicks (18 ± 3% of all blood mealson chicks) than on house sparrows (4.9 ± 3%). Birds ofboth species ate an average of 9.4 ± 1.2% of mosquitoes,and this percentage was positively correlated with defensivebehavior. High mosquito feeding success on house sparrows supportsits role as a potential amplifying host of West Nile virus.