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Effective management of insect disease vectors requires a detailed understanding of their ecology and behavior. In Anopheles gambiae sensu lato (s.l.) (Diptera: Culicidae) mating occurs during swarming, but knowledge of their mating behavior under natural conditions is limited. Mosquitoes mate in flight over specific landmarks, known as swarm markers, at particular locations. Swarms consist of males; the females usually approach the swarm and depart following copulation. The number of mating pairs per swarm is closely associated with swarm size. The shape and height of swarm markers vary and may depend on the environmental conditions at the swarm’s location. Male–male interactions in mosquito swarms with similar levels of attractive flight activity can offer a mating advantage to some individuals. Flight tone is used by mosquitoes to recognize the other sex and choose a desirable mate. Clarifying these and other aspects of mosquito reproductive behavior can facilitate the development of population control measures that target swarming sites. This review describes what is currently known about swarming behavior in Anopheles gambiae s.l., including swarm characteristics; mating within and outside of swarms, insemination in females, and factors affecting and stimulating swarming.

Culex quinquefasciatus, a member of the Culex pipiens complex, is widespread in Saudi Arabia and other parts of the world. It is a vector for lymphatic filariasis, Rift Valley fever, and West Nile virus. Studies have shown the deleterious effect of RNA interference (RNAi)-mediated knockdown of various lethal genes in model and agricultural pest insects. RNAi was proposed as a tool for mosquito control with a focus on Aedes aegypti and Anopheles gambiae. In this study, we examined the effect of RNAi of selected target genes on both larval mortality and adult emergence of Cx. quinquefasciatus through two delivery methods: soaking and nanoparticles. Ten candidate genes were selected for RNAi based on their known lethal effect in other insects. Disruption of three genes, chitin synthase-1, inhibitor of apoptosis 1, and vacuolar adenosine triphosphatase, resulted in the highest mortality among the selected genes using the two treatment methods. Silencing the other seven genes resulted in a medium to low mortality in both assays. These three genes are also active against a wide range of insects and could be used for RNAi-based mosquito control in the future.

Anopheles mosquitoes are the main vectors of malaria. There is little information on the current entomological aspects of Anopheles mosquitoes in Amhara region of northwestern Ethiopia. Therefore, the aim of this study was to assess the prevailing species composition, parous rate, and infection rate of Anopheles mosquitoes in the Bahir Dar city administration. A community-based cross-sectional study was conducted from January through July 2020. For this, six Centers for Disease Control and Prevention light traps (three traps indoor and three traps outdoor) were used to collect adult female Anopheles mosquitoes. The species were morphologically identified, and the parous and infection rates were determined via dissection of ovaries and salivary gland, respectively. A total of 378 adult female Anopheles mosquitoes comprised of three species (Anopheles d’thali, Anopheles rhodesiensis, and Anopheles gambiae complex) were collected and identified at the study sites. Anopheles rhodesiensis was the predominant species accounting for 90% of all collections at the Zenzelima site, followed by An. gambiae complex (6.5%). In contrast, An. gambiae complex was the predominant species at the Tis Abay site, comprising 94% of captures. The overall parous and infection rates were 35 (62.5%) and 1 (2.9%), respectively.

Vector control strategies recommended by the World Health Organization are threatened by resistance of Anopheles mosquitoes to insecticides. Information on the distribution of resistant genotypes of malaria vectors is increasingly needed to address the problem. Ten years of published and unpublished data on malaria vector susceptibility/resistance and resistance genes have been collected across Togo. Relationships between the spatial distribution of resistance status and environmental, socio-economic, and landscape features were tested using randomization tests, and calculating Spearman rank and Pearson correlation coefficients between mosquito mortality and different gridded values. Anopheles gambiae sensu lato was resistant to DDT, pyrethroids, and the majority of carbamates and organophosphates. Three sibling species were found (i.e., An. gambiae, Anopheles coluzzii, and Anopheles arabiensis) with four resistance genes, including kdr (L1014F, L1014S, and N1575Y) and ace1 (G119S). The most frequent resistance gene was L1014F. Overall, no association was found between the susceptibility/resistance status and environmental features, suggesting that evolution of resistance may be most closely related to extreme selection from local insecticide use. Nevertheless, further research is necessary for firm conclusions about this lack of association, and the potential role of landscape characteristics such as presence of crops and percentage of tree cover.

The carboxamide N,N-di-ethyl-meta-toluamide (DEET) is the most effective and widely used insect repellent today. However, drawbacks concerning the efficacy and the safety of the repellent have led to efforts to design new classes of insect repellents. Through quantitative structure–activity relationships, chemists have discovered two chemical groups of novel repellents: the acylpiperidines and the carboxamides, with the acylpiperidines generally more potent in biological assays. Although the exact mechanism of action of DEET and other repellents has not yet been thoroughly elucidated, previous research shows that the activity of insect odorant receptors are inhibited in the presence of repellents. The present electrophysiological study employs two-electrode voltage clamp with Xenopus laevis oocytes expressing AgOR2/AgOrco and AgOR8/AgOrco receptors to assess the effects of the novel repellents on Anopheles gambiae Giles (Insecta: Diptera: Culicidae) mosquito odorant receptors. The novel acylpiperidines and carboxamides reversibly inhibited (12–91%) odorant-evoked currents from both AgOR2/AgOrco and AgOR8/AgOrco receptors in a dose-dependent manner at all tested concentrations (30 μM to 1 mM). Furthermore, all the novel agents were more potent inhibitors of the receptors than DEET, with the acylpiperidines producing on average greater inhibition than the carboxamides. Interestingly, there was a correlation (r2 = 0.72) between the percentage inhibition of AgOR2/AgOrco receptor currents and protection times of the acylpiperidines. Our results add to existing evidence that the repellency of a compound is linked to its ability to disrupt the insect olfactory system and that the acylpiperidines could represent a class of more effective alternatives to the current gold standard, DEET.

Studies done in Bouaké (Côte d’Ivoire) about 20-yr ago reported that Anopheles gambiae s.l. Giles was the major malaria vector. The present study aimed to update these data and to identify the main vectors. Mosquitoes were collected in Allokokro and Petessou villages between June 2014 and December 2015 using the human landing catching method. Potential breeding sites of An. gambiae s.l. were identified in August and October 2014 and mapped using GPS. Anopheles species were morphologically and molecularly [polymerase chain reaction (PCR)] identified. Ovaries of female were dissected to determine the parity and infection with Plasmodium was detected in head and thorax by quantitative PCR. In Allokokro, the biting rate of An. gambiae s.s was significantly greater than Anopheles coluzzii, whereas, in Petessou, biting rates of both species were comparable. Plasmodium falciparum (Haemosporida: Plasmodiidae), Plasmodium malariae (Haemosporida: Plasmodiidae), and Plasmodium ovale (Haemosporida: Plasmodiidae) identified in both villages. The infection rates of An. gambiae s.s. and An. coluzzii were not significantly different. The entomological inoculation rate (EIR) of An. gambiae s.s. for P. falciparum was 9-fold greater than that of An. coluzzii in Allokokro; however, in Petessou, the EIRs of both species were comparable. In both village, An. gambiae s.s was responsible for P. falciparum and P. ovale transmission whereas An. coluzzii transmitted all three Plasmodium species.

A model is developed of malaria (Plasmodium falciparum) transmission in vector (Anopheles gambiae) and human populations that include the capacity for both clinical and parasite suppressing immunity. This model is coupled with a population model for Anopheles gambiae that varies seasonal with temperature and larval habitat availability. At steady state, the model clearly distinguishes uns hypoendemic transmission patterns from stable hyperendemic and holoendemic patterns of transmission. The model further distinguishes hyperendemic from holoendemic disease based on seasonality of infection. For hyperendemic and holoendemic transmission, the model produces the relationship between entomological inoculation rate and disease prevalence observed in the field. It further produces expected rates of immunity and prevalence across all three endemic patterns. The model does not produce mesoendemic transmission patterns at steady state for any parameter choices, leading to the conclusion that mesoendemic patterns occur during transient states or as a result of factors not included in this study. The model shows that coupling the effect of varying larval habitat availability with the effects of clinical and parasite-suppressing immunity is enough to produce known patterns of malaria transmission.

BackgroundLarge distribution of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) contributed to a significant decrease in malarial mortality. Unfortunately, large insecticide resistance in malaria vectors occurred and is a threat to the future use of these control approaches. The purpose of this study was to explore a new approach for vector control. Patches containing Bacillus thuringiensis var. israelensis (Bti) solubilized Cry toxins mixed with sugar were developed and tested in the laboratory with pyrethroid-resistant Anopheles gambiae s.l. using tunnel tests.MethodsMosquitoes were released at 6:00 p.m. into a large tunnel separated by a bed net, perforated with nine holes, from a smaller chamber with a guinea pig. Nine Bti sugar patches (BSPs) were attached to the bed net between the nine holes. Fourteen hours later (8:00 a.m.), mosquitoes were collected from the tunnel and the guinea pig chamber. Live females were kept in cups and were fed a sugar solution (5%) for 72 h and delayed mortality was followed. The results were reported as passing, blood fed and mortality rates.ResultsMosquito populations that are resistant to the insecticides in the bed net, exhibited high mortality (60%) in the presence of the BSPs. Untreated bed nets with patches in the tunnel test killed 66–95% of the mosquitoes that landed and untreated bed nets were superior to treated bed nets.ConclusionBSPs efficiently kill resistant mosquitoes that land on treated and untreated bed nets and thus could ultimately reduce the number of vector-borne malarial mosquitoes.

Pyrethroids and DDT are key insecticides in the control of malaria, yellow fever, and lymphatic filariasis vectors. Knockdown and metabolic resistance mechanisms have been proven to be important in determining the efficacy of insecticides. Here we investigated cytochrome P450 as a resistance mechanism in Anopheles gambiae Giles and Culex quinquefasciatus Say exposed to deltamethrin and DDT. Two- to three-days-old adult female mosquitoes were used for insecticide exposures and PBO synergistic assays using WHO standard guidelines, kits and test papers (DDT 4%, deltamethrin 0.05%, and PBO 4%). Polymerase chain reaction (PCR) assays were used for the identification of the species and for characterization of the kdr allele. Mortality at 24 h post-exposure was 18 and 17% in An. gambiae s.s. exposed to DDT and deltamethrin, respectively; 1 and 5% in Cx. quinquefasciatus exposed to DDT and deltamethrin respectively. Significant (P < 0.01) levels of susceptibility was recorded in mosquitoes pre-exposed to PBO, as KDT50 and 24 h of exposure ranged from 37.6 min to 663.4 min and 27 to 80%, respectively. Presence of a knockdown resistance allele was recorded in An. gambiae s.s., 22.5% for homozygote resistance and 7.5% for heterozygotes, while Cx. quinquefasciatus populations showed no kdr allele despite the high level of resistance to DDT and deltamethrin. Findings from this study indicated that cytochrome P450 mono-oxygenase expression is highly implicated in the resistance phenotype to DDT and pyrethroids in An. gambiae and Cx. quinquefasciatus in the study area.

Recent results of high-altitude windborne mosquito migration raised questions about the viability of these mosquitoes despite ample evidence that many insect species, including other dipterans, have been known to migrate regularly over tens or hundreds of kilometers on high-altitude winds and retain their viability. To address these concerns, we subjected wild Anopheles gambiae s.l. Giles mosquitoes to a high-altitude survival assay, followed by oviposition (egg laying) and blood feeding assays. Despite carrying out the survival assay under exceptionally harsh conditions that probably provide the lowest survival potential following high altitude flight, a high proportion of the mosquitoes survived for 6- and even 11-h assay durations at 120- to 250-m altitudes. Minimal differences in egg laying success were noted between mosquitoes exposed to high altitude survival assay and those kept near the ground. Similarly, minimal differences were found in the female’s ability to take an additional bloodmeal after oviposition between these groups. We conclude that similar to other high-altitude migrating insects, mosquitoes are able to withstand extended high-altitude flight and subsequently reproduce and transmit pathogens by blood feeding on new hosts.

An effective control of malaria vectors requires an extensive knowledge of mechanisms underlying the resistance-phenotypes developed by these vectors against insecticides. We investigated Anopheles gambiae mosquitoes from Benin and Togo for their intensity of insecticide resistance and we discussed the involvement of genotyped mechanisms in the resistance-phenotypes observed. Three- to five-day-old adult mosquitoes emerged from field and laboratory An. gambiae larvae were assayed using WHO tube intensity tests against various doses of deltamethrin: 1× (0.05%); 2× (0.1%); 5× (0.25%); 7.5× (0.375%) and those of pirimiphos-methyl: 0.5× (0.125%); 1× (0.25%). Members of An. gambiae complex were screened in field populations using polymerase chain reaction (PCR) assays. The presence of kdrR(1014F/1014S) and ace-1R(119S) mutations was also investigated using TaqMan and PCR-RFLP techniques, respectively. Anopheles gambiae from field were very resistant to deltamethrin, whereas KisKdr and AcerKdrKis strains displayed 100% mortality rates at 2× the diagnostic dose. In contrast, the field mosquitoes displayed a low resistance-intensity against 1× the diagnostic dose of pirimiphos-methyl, whereas AcerKis and AcerKdrKis strains showed susceptibility at 0.5× the diagnostic dose. Anopheles gambiae s.s., Anopheles coluzzii, and Anopheles arabiensis were identified. Allelic frequencies of kdrR (1014F) and ace-1R (119S) mutations in the field populations varied from 0.65 to 1 and 0 to 0.84, respectively. The field An. gambiae displayed high-resistance levels against deltamethrin and pirimiphos-methyl when compared with those of the laboratory An. gambiae-resistant strains. These results exhibit the complexity of underlying insecticide resistance mechanisms in these field malaria vectors.

The biophysical environment plays an important role in the spatio-temporal abundance and distribution of mosquitoes. This has implications for the spread of vectors and diseases they cause across diverse landscapes. Here, we assessed vector mosquito abundances in relation to large water bodies, from three malaria districts in a semi-arid environment. Furthermore, we explored thermal limits to activity of the dominant and most medically important malaria vector across malaria-endemic areas. Mosquitoes were trapped near permanent water bodies across different districts. Critical thermal limits (critical thermal-maxima and -minima) to activity of wild adults and 4th instar larvae Anopheles arabiensis (Diptera: Culicidae) were assessed. Our results showed that Anopheles spp. dominate mosquito communities across all three districts, but that their numbers were far greater in Okavango than in other regions. At the Okavango sites, the numbers of Anopheles spp. decreased with distance from main water source. Anopheles spp. sampled in this region comprised Anopheles gambiae (Giles,1902) and Anopheles funestus (Giles, 1900) species complexes, with the former dominating in numbers. Thermal activity assays showed An. arabiensis females had wider thermal tolerance windows than males while larval thermal activity limits differed significantly across space. These results confirm that the Okavango district should be prioritized for vector control measures. Moreover, intervention strategies should consider recommendations for proximity effects to large water bodies, given the differential risk associated with distance from water. The wider thermal window on female vectors has implications for possible future malaria transmission and diverse habitat utilization under changing environments.

Indoor residual spraying (IRS) was applied in addition to the use of long-lasting insecticidal nets in the South West in Burkina Faso, where Anopheles gambiae s.l. the major malaria vector was resistant to pyrethroids. This study was designed to evaluate the efficacy and residual life of bendiocarb (active ingredient) used for spraying on different wall surfaces (mud and cement). Cone bioassays were done monthly with the susceptible An. gambiae ‘Kisumu’ strain and the local wild populations to determine the duration for which insecticide was effective in killing mosquitoes. Cone bioassay data showed low efficacy and short persistence of bendiocarb applied on mud and cement walls, lasting 2 mo with the susceptible insectary strain and less than 1 mo with An. gambiae wild populations. In addition, WHO tube assays confirmed resistance of An. gambiae wild populations to 0.1% bendiocarb with mortality rates less than 80% in both study sites (sprayed and unsprayed sites). The pilot study of IRS with bendiocarb showed that the residual efficacy of bendiocarb was very short, and resistance to bendiocarb was confirmed in wild populations of An. gambiae s.l. Therefore, Ficam 80 WP was not suitable for IRS in this area due to the short residual duration related mainly to vectors resistance to bendiocarb. While waiting for innovative malaria control tool, alternative insecticide (organophosphate or neonicotinoid classes) or combinations of insecticides have to be used for insecticide resistance management in Burkina Faso.

Malaria, dengue, yellow fever, and the Zika and West Nile Viruses are major vector-borne diseases of humans transmitted by mosquitoes. According to the World Health Organization, over 80% of the world’s population is at risk of contacting these diseases. Insecticides are critical for mosquito control and disease prevention, and insect insecticide resistance is on the increase; new alternatives with potentially different modes of action from current chemistry are needed. During laboratory screening of industrial minerals for insecticide activity against Anopheles gambiae (Giles) (Diptera: Culicidae) we discovered a novel mechanical insecticide derived from volcanic rock (MIVR) with potential use as a residual spray. In modified WHO cone tests, the time to 50% mortality was 5 h under high-humidity conditions. MIVR treated surfaces demonstrated no mosquito repellency. In field studies where the mechanical insecticide was applied to wood using standard spray equipment and then placed under stilt homes in New Orleans, LA, the residual activity was >80% after 9 wk against Aedes aegypti (L.) (Diptera: Culicidae), Aedes albopictus (Skuse) (Diptera: Culicidae) and Culex quinquefasciatus (Say) (Diptera: Culicidae) (with similar efficacy to a positive chemical insecticide control). In scanning electron microcopy studies, the MIVR was transferred as particles mostly to the legs of the mosquito. This wettable powder made from volcanic rock is a mechanical insecticide representing a potential new mode of action different from current chemistry for mosquito control and is in commercial development under the trade name Imergard™WP as an indoor and outdoor residual spray.

Laboratory experiments intended to test the effects of a purported ovipositional deterrent pheromone of Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) led instead to the finding that egg cannibalism by fourth-instar larvae explained the 'deterrent effect' rather than chemical signals from larvae. Virtually no eggs were counted from cups containing fourth-instar larvae, after oviposition. Numbers of eggs from cups containing younger, conspecific larvae, or pupae did not differ significantly from water only control. Video recording of larval behavior revealed that fourth-instar larvae consumed entire eggs in one continuous ingestive bout. Egg output in the presence of fourth-instar larvae with their mouthparts sealed was similar to the larvae-absent treatment, but nearly 100-fold greater than that counted in the presence of fourth-instar larvae that could eat freely. Thus, the huge egg reduction in eggs counted in the presence of fourth-instar larvae was due to egg cannibalism, not because presence of these larvae deterred oviposition as has been reported in previous studies. Third and especially fourth-instar larvae also cannibalized newly hatched first-instar larvae. Artificial food availability did not eliminate but did reduce cannibalism during first several hours of exposure. The rate at which fourth-instar larvae consumed conspecifics rose significantly and linearly with prey density, generating a non-plateauing, type I functional response curve. These results clearly show that care must be taken such that mosquito ovipositional experiments are not confounded by presence of cannibalizing larvae.

The biting behavior of anophelines is an important determinant of malaria transmission. Understanding the local vector host-seeking behavior, its outdoor/ indoor biting preference, and nocturnal biting periods is essential for effectively applying and improving vector control methods, such as Long Lasting Insecticidal Nets (LLINs) and personal protective measures. To better understand the biting and host-seeking patterns of Anopheles mosquitoes in Northwestern Burkina Faso, we performed biweekly Human Landing Catches (HLC) in six villages during the period of highest mosquito abundance and malaria transmission. We applied a negative binomial regression framework to statistically analyze the host-seeking activities of Anopheles species and test for differences across hours, months, and villages, as well as for differences between indoor and outdoor capture points. Anopheles gambiae s.l. was identified as the main malaria vector in this region, representing about 90% of the total anopheline population. Biting activity was significantly different across hours and showed a peaked plateau between 2000 and 0200 hours. Differences in the pattern of biting cycles were observed between the early and late rainy season. This study shows that anopheline biting activity in Northwest Burkina Faso is high throughout the night, at indoor and outdoor posts alike. Consequently, bed nets alone may not provide sufficient protection against early biting anophelines and should be complemented with additional strategies such as indoor residual spraying (IRS) and larval source management (LSM) to meet the WHO’s ambitious goals that are reflected in the global technical malaria strategy for 2030.

Insecticidal indoor residual wall treatment is a major tool for the control of malaria, with the goals of reducing indoor vector density and vector life span, in addition to reducing transmission rates of disease. Dynamics of the malaria vector, Anopheles gambiae, in the Emutete region in the Western Kenya highlands are based on an already existing model in the literature. In this paper, the framework is used to predict vector reduction due to four types of indoor wall treatments: two cases of indoor residual spraying of DDT and two types of pyrethrin-based INESFLY insecticidal paint. These treatments differ primarily in the duration of their persistence on walls. The model shows the extent of suppression of vector abundance over time due to each of the four treatments. It predicts that indoor residual spraying may have no noticeable effect at all if the percent coverage is not high enough or the persistence of the mortality effect is low, but will have a substantial effect at higher coverage rates and/or higher persistence. For treatments with longer persistence of mortality, the model predicts a coverage threshold above which extra treatment has little to no effect. For treatments of short persistence of mortality, the seasonal timing of treatment has a noticeable effect on the duration of vector suppression. Overall, the model supports claims in the literature that wall treatments have the capacity to reduce the vector burden.

The population sizes of Anopheles gambiae Giles (Diptera: Culicidae) and Anopheles arabiensis Patton (Diptera: Culicidae) increase dramatically with the onset of the rainy season in sub-Saharan Africa, but the ecological mechanisms underlying the increases are not well understood. As a first step toward to understand, we investigated the proliferation of algae, the major food of mosquito larvae, in artificial fresh water bodies exposed to sunlight for a short period, and old water bodies exposed to sunlight for a long period, and the effects thereof on the development of these anopheline larvae. We found that an epizoic green algal species of the genus Rhopalosolen (Chlorophyta: Chlorophyceae) proliferated immediately after water freshly taken from a spring was placed in sunlight. This alga proliferated only briefly (for ~10 d) even if the water was repeatedly exposed to sunlight. However, various algal species were observed in water that remained under sunlight for 40 d or longer (i.e., in old water bodies). The growth performance of larvae was higher in sunlight-exposed (alga-rich) water than in shade-stored (alga-poor) water. Stable isotope analysis suggested that these two anopheline species fed on Rhopalosolen algae in fresh water bodies but hardly at all on other algae occurring in the old water bodies. We concluded that freshly formed ground water pools facilitate high production of anopheline species because of the proliferation of Rhopalosolen algae therein, and the increase in the number of such pools in the rainy season, followed by rapid increases in A. gambiae and A. arabiensis numbers.

The main malaria vectors in sub-Saharan Africa, the Anopheles gambiae (Giles; Diptera: Culicidae), normally breed in clean water sources. However, evidence suggests an on-going adaptation of Anopheline species to polluted breeding habitats in urban settings. This study aimed at understanding the adaptation to breeding in water bodies with different qualities, in five selected mosquito breeding sites in urban Accra, Ghana. The study sites were also evaluated for the WHO water-quality parameters as a measure of pollution, and insecticide residues. Field mosquitoes were evaluated for five genes; CYP6P3, CYP4H19, CYP4H24, GSTD1-4, and ABCC11-associated with insecticide detoxification-using quantitative RT-PCR, as well as Mono-oxygenase, Alpha Esterase, Glutathione S-transferase, and insensitive acetylcholinesterase (AChE) using biochemical enzyme assays. The lab-reared, insecticide susceptible An. gambiae Kisumu strain was bred in the most polluted water source for 10 generations and evaluated for the same genes and enzymes. The results revealed that the fold expression of the genes was higher in the larvae compared with the adults. The results also suggest that detoxification enzymes could be involved in the adaptation of An. gambiae to polluted breeding sites. Correlation analysis revealed a highly positive significant correlation between calcium levels and all five genes (P < 0.05). Stepwise linear regression to understand which of the variables predicted the expression of the genes revealed that sulphate was responsible for ABCC11 and CYP4H24, alkalinity for GSTD1-4, and calcium for CYP4H19 and CYP6P3. The detailed genetic basis of this adaptation need to be further investigated. A further understanding of this adaptation may provide outlooks for controlling malaria and other disease vectors adapted to polluted breeding water sources.

Insecticide decay rate on different wall surfaces is of importance to indoor residual spray (IRS) programs used as a malaria control intervention. Past IRS operations showed increasing populations of endophilic malaria vectors resting on indoor surfaces from various sites in Uganda following use of Ficam VC (bendiocarb) insecticide; variability of insecticide life was believed to be primarily due to wall surface type. Bendiocarb longevity was tested in the northern Uganda districts of Amuru, Apac, and Pader to assess its residual efficacy on three commonly encountered wall surfaces. Wall types included mud and wattle, plain brick, and painted plaster. A susceptible mosquito strain (Anopheles gambiae Kisumu) was used in all trials. Nine houses in each of the three districts were set with three test cones and one control cone per house, divided evenly among the three wall surfaces. Bioassays were run monthly through 6 mo. Painted plastered surfaces produced 100% mortality (at 24 h) through 6 mo. Plain brick surfaces killed 100% of test mosquitoes through 4 mo, while mud and wattle wall surfaces produced a 98% mortality rate at 3 mo post spray. The KD60 (knockdown at 60 min) for painted plastered surfaces was 100% for 6 mo, plain brick surface KD60 was 80% at 6 mo, and the mud and wattle surface KD60 was >80% at 3 mo. There was a significant effect on Ficam VC longevity by wall type and evidence of a relationship between test period and wall type on the KD60.

Insecticide resistance is one of the primary threats to the recent gains in malaria control. This is especially true in Guinea, where long-lasting insecticidal nets are currently the primary vector control intervention. To better inform the national malaria control program on the current status of insecticide resistance in Guinea, resistance bioassays were conducted, using Anopheles gambiae s.l. Giles, in three sites. Molecular analyses were also done on An. gambiae s.l. to determine the species and find whether the target-site mutations kdr and Ace1R were present. Susceptibility tests revealed resistance to DDT and pyrethroids, although mosquitoes were susceptible to deltamethrin in two of the three sites tested. Mosquitoes were susceptible to bendiocarb, except in Kissidougou, Guinea. The kdr-west mutation was widespread and the frequency was 60% or more in all sites. However, the Ace1R mutation was present in low levels. Insecticide susceptibility should continue to be monitored in Guinea to ensure insecticide-based vector control methods remain effective.

The scaling-up of malaria control interventions in northern Tanzania has resulted in a decline in malaria prevalence and vector species composition. Despite this achievement, residual malaria transmission remains a concern in the area. The main aim of this study was to investigate malaria vector species composition, parasite infectivity rates, and the presence of insecticide knockdown resistance (kdr) mutations in three sites that have experienced a significant decline in malaria in northern Tanzania. Adult mosquitoes were sampled using light traps in houses and hand-aspirators in cowsheds, whereas the standard dipping method was used for sampling mosquito larvae. Adult mosquitoes identified as Anopheles gambiae s.l. and An. funestus s.l. and larval stages III and IV of An. gambiae s.l. were stored in absolute ethanol for further laboratory molecular identification. The identified species in the An. gambiae complex were An. gambiae s.s., An. merus, An. quadriannulatus, and An. arabiensis, whereas the An. funestus group comprised An. funestus s.s., An. rivulorum, and An. leesoni. For An. gambiae s.s. analyzed from Zeneth, 47.6% were kdr-East homozygous susceptible, 35.7% kdr-East heterozygous resistant, 9.6% kdr-East homozygous resistant, and 7.1% undefined, whereas specimens from Kwakibuyu were 45.5% kdr-East homozygous susceptible, 32.7% kdr-East heterozygous resistant, 16.3% kdr-East homozygous resistant, and 5.5% undefined. There were no kdr-West alleles identified from any specimen. The overall malaria parasite infectivity rate was 0.75%. No infections were found in Moshi. The findings indicate that populations of the major malaria vector mosquitoes are still present in the study area, with An. funestus taking a lead in malaria transmission.

A survey of susceptibility to DDT, deltamethrin, bendiocarb, and chlorpyrifos-methyl was conducted in five localities in 2011 in Togo, West Africa, to assess the insecticide resistance status of Anopheles gambiae s.l. (Diptera: Culicidae). Female populations of An. gambaie s.l. emerged from collected larvae (F0) were exposed to insecticide-impregnated papers using World Health Organization test kits for adult mosquitoes; the susceptible reference strain Kisumu was used as a control. Resistance to DDT and deltamethrin was observed within the mosquito populations tested. Anopheles gambiae s.s. and Anopheles coluzzii represented the only species recorded in the study sites. The frequency of knockdown resistance (kdr L1014F) mutation determined using polymerase chain reaction diagnostic tests was lower in An. gambiae than in An. coluzzii in all of the localities except Kolokopé. Further investigations of An. gambiae s.l. resistance are needed in Togo to help the National Malaria Control Programme in vector control decision making and implementation of resistance management strategy.

Background: An insecticide susceptibility study was carried out on Anopheles gambiae sensu lato vectors in 11 districts in Ghana between October 2012 and January 2013. Methods: An. gambiae s.l. larvae were collected, bred under standard conditions, and 3–5 d postemerged females were used for bioassay. Between 22 and 25 female An. gambiae s.l. fed only 10% sugar were used for testing. Exposure was for 1 h (2 h for fenitrothion). An . gambiae that were knocked down were recorded every 10 min and mortalities recorded 24 h posttest. Eleven insecticides from four chemical classes were used: organochlorines, organophosphates, carbamates, and pyrethroids. Subsamples of An . gambiae were analyzed by polymerase chain reaction for species and knockdown resistance ( kdr ) allele determination. Results: Malathion was effective in killing An. gambiae in seven districts, fenitrothion in three districts, and propoxur in one district. The organophosphate and carbamate insecticides were effective in killing An. gambiae compared to pyrethroids and organochlorines. Of the limited samples analyzed, An. gambiae sensu stricto (39/110), An. coluzzii (66/110), and An. arabiensis (5/110) were identified. Few kdr (11/110) susceptible mosquitoes were detected. Homozygous kdr RR (65/110) and heterozygous kdr RS (8/110) genotypes were identified. Conclusions: An organophosphate insecticide is considered appropriate for indoor residual spraying (IRS) in the 11 districts earmarked for the IRS program in Ghana.

Mosquitoes derive energy from plant sugar, thereby promoting survival and reproduction. Its survival value to females plays a key role in the vectorial capacity of mosquito populations. Previous olfactometry assays of responsiveness demonstrated that Senna didymobotrya Fresenius, Parthenium hysterophorus, L. Senna occidentalis, (L) and Lantana camara L were among the most attractive plants for the Mbita strain of Anopheles gambiae s.s. Giles in eastern Africa. Here, we provide experimental evidence that three of these four species also provide varying but substantial amounts of sugar for mosquito survival, whereas a fourth does not. Rank order of survival of both sexes of mosquitoes housed with these plants was as follows: S. didymobotrya was highest, followed by S. occidentalis and L. camara, whereas survival on P. hysterophorus was only slightly better than on only water. A positive control group, housed with 10% sucrose, survived well but fell significantly short of those with S. didymobotrya. A causal connection between survival and sugar availability was established by exposing mosquitoes to plants overnight, and then testing them for the presence and amount of undigested fructose. Fructose positivity was most frequent in those exposed to L. camara, whereas greatest amounts of fructose were obtained from S. occidentalis and S. didymobotrya. Parthenium hysterophorus scored lowest in both categories. We conclude that attractiveness and sugar availability are often, but not always, concordant. It remains unclear why P. hysterophorus should be attractive if it offers little sugar and does not prolong survival. Furthermore, the cause behind the superior survival benefit of S. didymobotrya, compared with 10% sucrose, is unknown.

Comprehensive knowledge on vector dynamics is lacking in Botswana and yet essential for effective indoor residual spraying. This study assessed some of the entomological indices that contribute to malaria transmission by an indoor-resting population of Anopheles arabiensis Patton (Diptera: Culicidae) in Tubu village, Okavango subdistrict. The pyrethroid space-spray technique and hut searches were used to sample mosquitoes. Species and bloodmeal source identification were done using the polymerase chain reaction techniques. The infective status was determined by the enzyme-linked immuno-sorbent assay test. The human blood indices (HBI), human-biting rates (HBR), and vector densities were computed. Anopheles arabiensis was the sole vector and member of the Anopheles gambiae Giles complex identified. Significant changes in vector densities were observed over seasons, while nonsignificant differences were observed among the huts (P > 0.05). The main source of bloodmeal was cattle (46.8% [65]). There were no significant differences in HBI (P > 0.05) and HBR (P > 0.05) between the seasons. All the 271 mosquitoes tested for sporozoite infection were negative. The results showed that seasonal variations in vector densities corresponded to the traditional malaria and nonmalaria seasons. The vector population was relatively more zoophagic. The HBI and HBR were not influenced by the seasons. None of the endophilic mosquitoes tested (N = 271) was positive for sporozoites. Our study provided some relevant basic information to the local malaria control program that may be used to strategize their operations if malaria elimination has to be achieved by 2017.

Aedes aegypti and Anopheles gambiae are two mosquito species that represent significant threats to global public health as vectors of Dengue virus and malaria parasites, respectively. Although mosquito populations have been effectively controlled through the use of synthetic insecticides, the emergence of widespread insecticide-resistance in wild mosquito populations is a strong motivation to explore new insecticidal chemistries. For these studies, Ae. aegypti and An. gambiae were treated with commercially available plant essential oils via topical application. The relative toxicity of each essential oil was determined, as measured by the 24-h LD(50) and percentage knockdown at 1 h, as compared with a variety of synthetic pyrethroids. For Ae. aegypti, the most toxic essential oil (patchouli oil) was ∼1,700-times less toxic than the least toxic synthetic pyrethroid, bifenthrin. For An. gambiae, the most toxic essential oil (patchouli oil) was ∼685-times less toxic than the least toxic synthetic pyrethroid. A wide variety of toxicities were observed among the essential oils screened. Also, plant essential oils were analyzed via gas chromatography/mass spectrometry (GC/MS) to identify the major components in each of the samples screened in this study. While the toxicities of these plant essential oils were demonstrated to be lower than those of the synthetic pyrethroids tested, the large amount of GC/MS data and bioactivity data for each essential oil presented in this study will serve as a valuable resource for future studies exploring the insecticidal quality of plant essential oils.

Members of the Anopheles gambiae sensu lato (Giles) complex define a group of seven morphologically indistinguishable species, including the principal malaria vectors in Sub-Saharan Africa. Members of this complex differ in behavior and ability to transmit malaria; hence, precise identification of member species is critical to monitoring and evaluating malaria threat levels. We collected mosquitoes from five counties in Liberia every other month from May 2011 until May 2012, using various trapping techniques. A. gambiae complex members were identified using molecular techniques based on differences in the ribosomal DNA (rDNA) region between species and the molecular forms (S and M) of A. gambiae sensu stricto (s.s) specimens. In total, 1,696 A. gambiae mosquitoes were collected and identified. DNA was extracted from legs of each specimen with species identification determined by multiplex polymerase chain reaction using specific primers. The molecular forms (M or S) of A. gambiae s.s were determined by restriction fragment length polymorphism. Bivariate and multivariate logistic regression models identified environmental variables associated with genomic differentiation. Our results indicate widespread occurrence of A. gambiae s.s., the principal malaria vector in the complex, although two Anopheles melas Theobald/A. merus Donitz mosquitoes were detected. We found 72.6, 25.5, and 1.9% of A. gambiae s.s specimens were S, M, and hybrid forms, respectively. Statistical analysis indicates that the S form was more likely to be found in rural areas during rainy seasons and indoor catchments. This information will enhance vector control efforts in Liberia.

We understand little about photo-preference and the molecular mechanisms governing vision-dependent behavior in vector mosquitoes. Investigations of the influence of photo-preference on adult mosquito behaviors such as endophagy and exophagy and endophily and exophily will enhance our ability to develop and deploy vector-targeted interventions and monitoring techniques. Our laboratory-based analyses have revealed that crepuscular period photo-preference differs between An. gambiae and An. stephensi. We employed qRT-PCR to assess crepuscular transcriptional expression patterns of long wavelength-, short wavelength-, and ultraviolet wavelength-sensing opsins (i.e., rhodopsin-class G-protein coupled receptors) in An. gambiae and in An. stephensi. Transcript levels do not exhibit consistent differences between species across diurnal cycles, indicating that differences in transcript abundances within this gene set are not correlated with these behavioral differences. Using developmentally staged and gender-specific RNAseq data sets in An. gambiae, we show that long wavelength-sensing opsins are expressed in two different patterns (one set expressed during larval stages, and one set expressed during adult stages), while short wavelength- and ultraviolet wavelength-sensing opsins exhibit increased expression during adult stages. Genomic organization of An. gambiae opsins suggests paralogous gene expansion of long wavelength-sensing opsins in comparison with An. stephensi. We speculate that this difference in gene number may contribute to variation between these species in photo-preference behavior (e.g., visual sensitivity).

Surveillance of malaria vectors in Africa is most often accomplished using CDC-type light traps or human landing catches (HLCs). Over the past 30 yr, a variety of commercial and experimental mosquito traps have been developed for residential mosquito control or for improved surveillance of disease vector species, including the BG Sentinel (BGS) trap. To optimize collection of Anopheles gambiae Giles using this trap, BGS traps were modified with an opening (vent) added to the trap base to decrease exhaust airflow. Four traps configurations were tested with colony-reared host-seeking female An. gambiae in free-flying laboratory enclosures. Six attractant treatments (three attractants: BG-Lure, Limburger cheese, and a blank, with and without CO2) were tested concurrently. Across all trap-attractant combinations, significantly more mosquitoes (P < 0.05) were collected in standard, unmodified traps set upright (mean: 10.0) or upside down (9.8) than vented traps, whether upright (5.9) or upside down (7.0). The BG-Lure + CO2 and Limburger Cheese + CO2 bait combinations were more attractive than the other attractants tested alone. All attractant combinations collected significantly more mosquitoes than unbaited traps. Field studies are needed to determine if BG-Lure + CO2- or Limburger cheese + CO2-baited BGS traps are comparable with HLCs in collecting host-seeking An. gambiae.

Impoundments formed by microdams in rural areas of Africa are important sources of water for people, but they provide potential larval habitats for Anopheles (Diptera: Culicidae) mosquitoes that are vectors of malaria. To study this association, the perimeters of 31 microdam impoundments in western Kenya were sampled for Anopheles larvae in three zones (patches of floating and emergent vegetation, shorelines of open water, and aggregations of cattle hoofprints) across dry and rainy seasons. Of 3,169 larvae collected, most (86.8%) were collected in the rainy season. Of 2,403 larvae successfully reared to fourth instar or adult, nine species were identified; most (80.2%) were Anopheles arabiensis Patton, sampled from hoofprint zones in the rainy season. Other species collected were Anopheles coustani Laveran, Anopheles gambiae s.s. Giles, Anopheles funestus Giles, and Anopheles rivulorum Leeson, Anopheles pharoensis Theobald, Anopheles squamosus Theobald, Anopheles rufipes (Gough), and Anopheles ardensis (Theobald). Larvae of An. funestus were uncommon (1.5%) in both dry and rainy seasons and were confined to vegetated zones, suggesting that microdam impoundments are not primary habitats for this important vector species, although microdams may provide a dry season refuge habitat for malaria vectors, contributing to population persistence through the dry season. In this study, microdam impoundments clearly provided habitat for the malaria vector An. arabiensis in the rainy season, most of which was within the shallow apron side of the impoundments where people brought cattle for watering, resulting in compacted soil with aggregations of water-filled hoofprints. This observation suggests a potential conflict between public health concerns about malaria and people’s need for stable and reliable sources of water.

Changes in spatial distribution of mosquitoes over time in a Sahelian village were studied to understand the sources of the mosquitoes during the dry season when no larval sites are found. At that time, the sources of Anopheles gambiae Giles may be local shelters used by aestivating mosquitoes or migrants from distant populations. The mosquito distribution was more aggregated during the dry season, when few houses had densities 7- to 24-fold higher than expected. The high-density houses during the dry season differed from those of the wet season. Most high-density houses during the dry season changed between years, yet their vicinity was rather stable. Scan statistics confirmed the presence of one or two adjacent hotspots in the dry season, usually found on one edge of the village. These hotspots shifted between the early and late dry season. During the wet season, the hotspots were relatively stable near the main larval site. The locations of the hotspots in the wet season and early and late dry season were similar between years. Season-specific, stable, and focal hotspots are inconsistent with the predictions based on the arrival of migrants from distant localities during the dry season, but are consistent with the predictions based on local shelters used by aestivating mosquitoes. Targeting hotspots in Sahelian villages for vector control may not be effective because the degree of aggregation is moderate, the hotspots are not easily predicted, and they are not the sources of the population. However, targeting the dry-season shelters may be highly cost-effective, once they can be identified and predicted.

Alphaviruses are arthropod-borne pathogens that infect a range of hosts. In humans and other mammals, alphavirus infection can cause severe disease. In mosquito hosts, however, there are generally few symptoms. Little is known about the cellular responses of mosquitoes that allow them to cope with infection. In this investigation, a six-plex tandem mass tagging proteomic approach was used to study protein accumulation changes in the midgut of Anopheles gambiae (Giles) (Diptera: Culicidae) mosquitoes infected with o'nyong-nyong virus (Togaviridae, Alphavirus). Five hundred thirty-six nonredundant proteins were identified. Twenty-two were found in significantly different quantities in infected midguts compared with controls. Of interest, analysis revealed molecular pathways possibly targeted by virus proteins, such as those involving TAF4 and DNA polymerase phi proteins. Also identified was an FK506-binding protein. FK506-binding protein orthologs have been described as conserved host resistance factors, which suppress dengue and West Nile virus infection in human HeLa cells. This investigation constitutes the first study of the midgut-specific proteome of An. gambiae in relation to alphavirus infection. Our findings offer insight into mosquito immunity, including factors that possibly contribute to the different pathological outcomes observed in vertebrate and insect hosts.

The primary malaria vector in sub-Saharan Africa, Anopheles gambiae Giles (Diptera: Culicidae), is an anthropophagic and endophilic mosquito targeted for control with insecticides applied to interior resting surfaces and impregnated onto bed net materials. Effective malaria vector management involves monitoring the insecticide susceptibility of mosquito populations. Contemporary bioassays are based on mosquito contact with insecticide residues. We developed an innovative insecticide bioassay system that involves mosquito ingestion of a sugar-insecticide solution. The sucrose—permethrin solution in our bioassay system contained Trypan blue dye, creating a visual marker of insecticide ingestion in the mosquito's abdomen. Blue fecal spots deposited in the bioassay container provided further evidence of mosquito feeding. We used our bioassay to characterize the permethrin susceptibility of adult females of two strains of A. gambiae, one of which was susceptible and the other exhibited reduced suscept...

Before the release of genetically-modified or sterile male mosquitoes in an attempt to control local populations of malaria vectors, it is crucial to determine male traits involved in mating success. The effects of male size and age as determinants of male mating success in Anopheles gambiae s.s. were measured in the field and under laboratory conditions in Burkina Faso. First, the body sizes (estimated by wing length) of mating, swarming, and indoor-resting male mosquitoes were compared over a 3-yr period (2006–2009) from July to October in Soumousso and Vallee du Kou, two villages in western Burkina Faso. Second, the age structure of swarming and resting male mosquitoes were characterized based on the number of spermatocysts and the proportion of sperm in the reservoir of wild-caught male testis. Third, male age effects on the insemination rate of female An. gambiae were investigated in the laboratory. The mean size of males collected in copula was significantly larger than the mean for swarming males and indoor-resting males. The optimum male age for successful insemination of females was 4–8 d. These results suggest that male size is an important trait in determining male mating competitiveness in the field. Although age was not found to be a significant factor in mating competitiveness, it was significantly correlated with swarming behaviors in the field and insemination success in the laboratory. The implications of these results in terms of sexual selection in An. gambiae and vector control programs are further discussed.

Larval management of the malaria vector, Anopheles gambiae Giles s.s., has been successful in reducing disease transmission. However, pesticides are not affordable to farmers in remote villages in Mali, and in other material resource poor countries. Insect resistance to insecticides and nontarget toxicity pose additional problems. Neem (Azadirachta indica A. Juss) is a tree with many beneficial, insect bioactive compounds, such as azadirachtin. We tested the hypothesis that neem leaf slurry is a sustainable, natural product, anopheline larvicide. A field study conducted in Sanambele (Mali) in 2010 demonstrated neem leaf slurry can work with only the available tools and resources in the village. Laboratory bioassays were conducted with third instar An. gambiae and village methods were used to prepare the leaf slurry. Experimental concentration ranges were 1,061-21,224 mg/L pulverized neem leaves in distilled water. The 50 and 90% lethal concentrations at 72 h were 8,825 mg/L and 15,212 mg/L, respectively. LC concentrations were higher than for other parts of the neem tree when compared with previous published studies because leaf slurry preparation was simplified by omitting removal of fibrous plant tissue. Using storytelling as a medium of knowledge transfer, villagers combined available resources to manage anopheline larvae. Preparation of neem leaf slurries is a sustainable approach which allows villagers to proactively reduce mosquito larval density within their community as part of an integrated management system.

A study was conducted to investigate the current resting behavior of malaria vectors in Mutare and Mutasa districts, Zimbabwe. Mosquitoes were captured using pyrethrum spray collection, prokopac aspirator, pit shelter, and exit trap methods. Mosquitoes were sorted and identified using morphological key and polymerase chain reaction (PCR) techniques. The Anopheles funestus group constituted 97%, whereas Anopheles gambiae complex mosquitoes were few (3%). Endophilic collections in both species were five times greater than exophilic catches. The endophilic trait was further demonstrated by gravid to fed index (gravid/fed) of constantly more than 1. Nearly 90% endophilic An. funestus populations were collected on sprayable and 10% collected on unsprayable surfaces. Of the sprayable surfaces, 56% were collected on the roofs, with 44% on the walls. Of those on the walls, 44, 22, and 34% were caught on wall heights >1, 1.0–1.5

Pyrethroid-resistant malaria vectors have become a serious threat for malaria control, and bed nets that reduce the development of resistance are urgently needed. Here, we tested the effects of bed nets treated with the insect growth regulator pyriproxyfen against adult female Anopheles gambiae Giles (Diptera: Culicidae) under laboratory conditions. Noninsecticidal nets made of 195 denier monofilament polyethylene with a mesh size of 75 holes per square inch (equivalent to the Olyset Net) were dipped in a 0.1, 0.01, or 0.001% (wt:vol) alcohol solution of pyriproxyfen and dried overnight. Adult females of an insecticide-susceptible An. gambiae strain were exposed to treated and untreated nets before and after a bloodmeal. Bioassays showed that females were completely sterilized after exposure to 0.1% (35 mg [AI]/m2) and 0.01% pyriproxyfen-treated nets both before and after a bloodmeal. In addition, adult longevity decreased after exposure to the pyriproxyfen-treated nets in a concentration-dependent manner. The sterilizing and life-shortening effects of pyriproxyfen on the vector mosquito indicate that the combined use of pyriproxyfen and pyrethroids on bed nets has the potential to provide better malaria control and prevent the further development of pyrethroid resistance in malaria vectors.

A dynamical model of Anopheles gambiae larval and adult populations is constructed that matches temperature-dependent maturation times and mortality measured experimentally as well as larval instar and adult mosquito emergence data from field studies in the Kenya Highlands. Spectral classification of high-resolution satellite imagery is used to estimate household density. Indoor resting densities collected over a period of one year combined with predictions of the dynamical model give estimates of both aquatic habitat and total adult mosquito densities. Temperature and precipitation patterns are derived from monthly records. Precipitation patterns are compared with average and extreme habitat estimates to estimate available aquatic habitat in an annual cycle. These estimates are coupled with the original model to produce estimates of adult and larval populations dependent on changing aquatic carrying capacity for larvae and changing maturation and mortality dependent on temperature. This paper offers a general method for estimating the total area of aquatic habitat in a given region, based on larval counts, emergence rates, indoor resting density data, and number of households.Altering the average daily temperature and the average daily rainfall simulates the effect of climate change on annual cycles of prevalence of An. gambiae adults. We show that small increases in average annual temperature have a large impact on adult mosquito density, whether measured at model equilibrium values for a single square meter of habitat or tracked over the course of a year of varying habitat availability and temperature.

One strategy suggested for the management of mosquito insecticide resistance consists of combining a pyrethroid with an insecticide that has a different mode of action. To restore the efficacy of deltamethrin (pyrethroid) against pyrethroid-resistant strain of Anopheles gambiae Giles (VKPR: homozygous Kdr), deltamethrin was combined with the neonicotinoid insecticide dinotefuran and piperonyl butoxide (PBO). Bednets impregnated with deltamethrin, dinotefuran, and PBO alone and in combination were tested in the laboratory. Knockdown (KD) and mortality were measured using WHO cone tests on susceptible and pyrethroid-resistant adult mosquitoes. The combination of deltamethrin and PBO was synergistic against resistant female An. gambiae (58.2% mortality). Both mortality and knockdown time (KDt(50/95) values) of the tricomponent mixture on the VKPR strain were similar to the insecticidal activity of deltamethrin on the pyrethroid-susceptible KIS strain (98.8 and 100% mortality, respectively). The three-compound mixture of deltamethrin + PBO + dinotefuran showed an insecticidal efficacy greater than the deltamethrin + PBO mixture to the extent of completely restoring the efficacy of deltamethrin on pyrethroid-resistant An. gambiae.

Insecticide resistance in the main malaria vectors in Africa is a major concern for malaria vector control program managers. The most common insecticides used for indoor residual spraying (IRS) and treating bed nets are becoming increasingly ineffective. The quest for safer and more effective insecticides for malaria vector control is urgent. This study sought to evaluate the efficacy of ACTELLIC 50 EC (pirimiphos methyl), an organophosphate, for IRS in Ghana, where there is high vector resistance to pyrethroids and organochlorines. Before the commencement of the study, standard World Health Organization (WHO) vector susceptibility tests against a common malaria vector, Anopheles gambiae s.l, were conducted using preparations of pyrethroids, organochlorines, carbamates, and organophosphates. The vector was found to be resistant to the pyrethroids, the organochlorines, and the carbamates, but susceptible to the organophosphates. The emulsifiable organophosphate concentrate formulation, ACTELLIC 50 EC, was then evaluated to determine the efficacy and the length of its residual effect. The wall bioassay test, using recommended cones from WHO, was conducted on sprayed surfaces with ACTELLIC 50 EC from 27 July 2009 to 16 October 2009. After 15 wk of trials on painted cement surface, it was found out that the main malaria vector, An. gambiae s.l, was susceptible to the insecticide even though the WHO Pesticide Evaluation recommends 2- to 3-mo duration of effective action. Therefore, it is recommended for use in IRS programs in this part of Ghana, where there is high vector resistance to most of the insecticides.

Mosquito resistance to chemical insecticides is considered a serious threat for the sustainable use of contemporary malaria vector control methods. Fungal entomopathogens show potential as alternative biological control agents against (insecticide-resistant) anophelines. This study was designed to test whether the fungus, Beauveria bassiana, could be delivered to mosquitoes on netting materials that might be used in house screens, such as eave curtains. Tests were conducted to determine effects of formulation, application method, netting material, and nature of mosquito contact. Beauveria had a twice as high impact on Anopheles gambiae s.s. longevity when suspended in Shellsol solvent compared with Ondina oil (HR = 2.12, 95% confidence interval = 1.83-2.60, P < 0.001), and was significantly more infective when applied through spraying than dipping. Polyester and cotton bednets were the most effective substrates for mosquito infections, with highest spore viability on cotton nets. Whereas fungal impact was highest in mosquitoes that had passed through large-meshed impregnated nets, overall efficacy was equal between small- and large-meshed nets, with < or = 30-min spore contact killing >90% of mosquitoes within 10 d. Results indicate that the use of fungal spores dissolved in Shellsol and sprayed on small-meshed cotton eave curtain nets would be the most promising option for field implementation. Biological control with fungus-impregnated eave curtains could provide a means to target host-seeking mosquitoes upon house entry, and has potential for use in integrated vector management strategies, in combination with chemical vector control measures, to supplement malaria control in areas with high levels of insecticide resistance

Insecticidal properties of natural products may present alternatives to the use of synthetic molecule pesticides that are of diminishing effectiveness due to resistance. Three compounds, thymoquinone, nootkatone, and carvacrol, components of Alaska yellow cedar, Chamaecyyparis nootkatensis (D. Don) Spach, and incense cedar, Calocedrus decurrens (Torr.), essential oils, have been shown to have biological activity against a variety of mosquito and tick species. Although these components act as both repellents and insecticides, how they function in either capacity is unknown. Their use as mosquito control insecticides would be greatly increased if their mode of action is not the same as that of currently used commercial products. This study compared the lethal dosages for nootkatone, carvacrol, and thymoquinone by using colony strains of Anopheles gambiae Giles with known mutations at three different target sites. The altered target sites evaluated were the sodium channel para-locus mutation (L1014 F KDR) that confers permethrin resistance, the ACE-1 gene that confers organophosphate and carbamate resistance, and a gamma-aminobutyric acid receptor mutation of the Rdl locus conferring dieldrin resistance. Significant increases in lethal dose were not observed in any of the mosquito strains for any of the compounds tested compared with the doses required of chemicals with known modes of action at the mutated sites. Although the mode of action was not determined, this screening study indicates that none of these compounds interact at the target sites represented in the test mosquito strains. These compounds represent a different mode of action than existing chemicals currently used in mosquito control.

Reduced survival and future reproduction due to of current reproduction is a trade-off known as the cost of reproduction. Surprisingly, only a few studies have assessed the cost of reproduction in arthropod disease vectors, despite its effect on longevity, and thus on vectorial capacity. We evaluated the cost of reproduction on survival of Anopheles gambiae Giles by comparing mosquitoes that were denied exposure to the other sex, hereafter named virgins, and those that were allowed exposure to the other sex and mating, hereafter named mated. Merely 6 d of exposure to females with mating activity reduced male survival from a median of 17 d in virgins to 15 d in mated, indicating that male mating cost is substantial. The increase in mortality of mated males began several days after the exposure to females ended, indicating that mating is not associated with immediate mortality risk. Notably, body size was negatively correlated with male mortality in mated males, but not in virgins. The rate of insemination declined after 4 d of exposure to females, indicating that male mating capacity is limited and further supporting the hypothesis that mating is costly for males. Consistent with previous studies, female survival on sugar alone (median = 16 d) was shorter than on blood and sugar (median = 19 d), regardless if she was mated or virgin. Overall, survival of mated females was lower than that of virgins on a diet of blood and sugar, but no difference was found on a diet of sugar only. However, the cost of reproduction in females remains ambiguous because the difference in survival between virgin and mated females was driven by the difference between virgin (median = 19 d) and uninseminated females exposed to males (median = 17 d), rather than between virgin and inseminated females (median = 19 d). Accordingly, sperm and seminal fluid, egg development, and oviposition have negligible cost in terms of female survival. Only exposure to males without insemination decreased female survival. Nonetheless, if exposure to males under natural conditions is also associated with reduced survival, it might explain why females remain monogamous.