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This study explores the influence of small dams on the exposure to malaria vectors during the dry season in Kasungu district, Malawi, an area recently identified as high priority for malaria interventions by the National Malaria Control Programme. Small dam impoundments provide communities with a continuous supply of water for domestic and agricultural activities across sub‐Saharan Africa and are considered vital to food security and climate change resilience. However, these permanent water bodies also create ideal breeding sites for mosquitoes in typically arid landscapes. The study focuses on a specific dam impoundment and its vicinity, aiming to assess its spatial and temporal influence on indoor vector densities. From May to August 2021, CDC light traps were used to measure indoor mosquito densities for two consecutive nights per month in three communities located at increasing distances from the dam (0, ~1 and ~2 km). Simultaneously, drone imagery was captured for each community, enabling the identification of additional standing water within approximately 400 m of selected households. Larval sampling was carried out within the impoundment periphery and in additional water bodies identified in the drone imagery. Generalised linear mixed models (GLMMs) were employed to analyse the indoor Anopheles abundance data, estimating the effects of household structure (open/closed eaves), month, temperature and water proximity on malaria vector exposure. Throughout 685 trapping nights, a total of 1256 mosquitoes were captured, with 33% (412) being female Anopheles. Among these, 91% were morphologically identified as Anopheles funestus s.l., and 5% as Anopheles gambiae s.l. Catches progressively decline in each consecutive trapping month as the environment became drier. This decline was much slower in Malangano, the community next to the dam, with abundance being notably higher in June and July. Further, the majority of An. gambiae s.l. were caught in May, with none identified in July and August. Anopheles larvae were found both in the impoundment and other smaller water bodies such as irrigation wells in each survey month; however, the presence of these smaller water bodies did not have a significant impact on adult female mosquito catches in the GLMM. The study concludes that proximity to the dam impoundment was the primary driver of differences between survey communities with the abundance in Chikhombwe (~1 km away) and Chiponde (~2 km away) being 0.35 (95% confidence interval [CI], 0.19–0.66) and 0.28 (95% CI, 0.16–0.47) lower than Malangano, respectively, after adjusting for other factors. These findings underscore the importance of targeted interventions, such as larval source management or housing improvements, near small dams to mitigate malaria transmission risks during the dry season. Further research is needed to develop cost‐effective strategies for vector control within and around these impoundments. [ABSTRACT FROM AUTHOR]

There has been significant progress in malaria control in the last 2 decades, with a decline in mortality and morbidity. However, these gains are jeopardised by insecticide resistance, which negatively impacts the core interventions, such as insecticide‐treated nets (ITN) and indoor residual spraying (IRS). While most malaria control and research efforts are still focused on Anopheles gambiae complex mosquitoes, Anopheles funestus remains an important vector in many countries and, in some cases, contributes to most of the local transmission. As countries move towards malaria elimination, it is important to ensure that all dominant vector species, including An. funestus, an important vector in some countries, are targeted. The objective of this review is to compile and discuss information related to A. funestus populations' resistance to insecticides and the mechanisms involved across Africa, emphasising the sibling species and their resistance profiles in relation to malaria elimination goals. Data on insecticide resistance in An. funestus malaria vectors in Africa were extracted from published studies. Online bibliographic databases, including Google Scholar and PubMed, were used to search for relevant studies. Articles published between 2000 and May 2023 reporting resistance of An. funestus to insecticides and associated mechanisms were included. Those reporting only bionomics were excluded. Spatial variation in species distribution and resistance to insecticides was recorded from 174 articles that met the selection criteria. It was found that An. funestus was increasingly resistant to the four classes of insecticides recommended by the World Health Organisation for malaria vector control; however, this varied by country. Insecticide resistance appears to reduce the effectiveness of vector control methods, particularly IRS and ITN. Biochemical resistance due to detoxification enzymes (P450s and glutathione‐S‐transferases [GSTs]) in An. funestus was widely recorded. However, An. funestus in Africa remains susceptible to other insecticide classes, such as organophosphates and neonicotinoids. This review highlights the increasing insecticide resistance of An. funestus mosquitoes, which are important malaria vectors in Africa, posing a significant challenge to malaria control efforts. While An. funestus has shown resistance to the recommended insecticide classes, notably pyrethroids and, in some cases, organochlorides and carbamates, it remains susceptible to other classes of insecticides such as organophosphates and neonicotinoids, providing potential alternative options for vector control strategies. The study underscores the need for targeted interventions that consider the population structure and geographical distribution of An. funestus, including its sibling species and their insecticide resistance profiles, to effectively achieve malaria elimination goals. [ABSTRACT FROM AUTHOR]

In vertebrates, enzymes responsible for DNA methylation, one of the epigenetic mechanisms, are encoded by genes falling into the cytosine methyltransferases genes family (Dnmt1, Dnmt3a,b and Dnmt3L). However, in Diptera, only the methyltransferase Dnmt2 was found, suggesting that DNA methylation might act differently for species in this order. Moreover, genes involved in epigenetic dynamics, such as Ten‐eleven Translocation dioxygenases (TET) and Methyl‐CpG‐binding domain (MBDs), present in vertebrates, might play a role in insects. This work aimed at investigating nucleic acids methylation in the malaria vector Anopheles gambiae (Diptera: Culicidae) by analysing the expression of Dnmt2, TET2 and MBDs genes using quantitative real‐time polymerase chain reaction (qRT‐PCR) at pre‐immature stages and in reproductive tissues of adult mosquitoes. In addition, the effect of two DNA methylation inhibitors on larval survival was evaluated. The qPCR results showed an overall low expression of Dnmt2 at all developmental stages and in adult reproductive tissues. In contrast, MBD and TET2 showed an overall higher expression. In adult mosquito reproductive tissues, the expression level of the three genes in males' testes was significantly higher than that in females' ovaries. The chemical treatments did not affect larval survival. The findings suggest that mechanisms other than DNA methylation underlie epigenetic regulation in An. gambiae. [ABSTRACT FROM AUTHOR]

Understanding the environmental factors affecting the microbiota in malaria vectors may help in the development of novel vector control interventions, similar to paratransgenesis. This study evaluated seasonal and geographical variations in the microbial community of the two major malaria vectors. Adult Anopheles mosquitoes were collected across two different eco‐geographical settings in Cameroon, during the dry and wet seasons. DNA was extracted from the whole individual mosquitoes from each group and processed for microbial analysis using Illumina Miseq sequencing of the V3‐V4 region of the 16S rRNA gene. Data analysis was performed using QIIME2 and R software programs. A total of 1985 mosquitoes were collected and among them, 120 were selected randomly corresponding to 30 mosquitoes per season and locality. Overall, 97 bacterial taxa were detected across all mosquito samples, with 86 of these shared between dry and wet seasons in both localities and species. There were significant differences in bacterial composition between both seasons, with a clear separation observed between the dry and wet seasons (PERMANOVA comparisons of beta diversity, Pseudo‐F = 10.45; q‐value = 0.01). This study highlights the influence of seasonal variation on microbial communities and this variation's impact on mosquito biology and vectorial capacity should be further investigated. [ABSTRACT FROM AUTHOR]

The endosymbiont Wolbachia can have major effects on the reproductive fitness, and vectorial capacity of host insects and may provide new avenues to control mosquito‐borne pathogens. Anopheles gambiae s.l is the major vector of malaria in Africa but the use of Wolbachia in this species has been limited by challenges in establishing stable transinfected lines and uncertainty around native infections. High frequencies of infection of Wolbachia have been previously reported in An. gambiae collected from the Valle du Kou region of Burkina Faso in 2011 and 2014. Here, we re‐evaluated the occurrence of Wolbachia in natural samples, collected from Valle du Kou over a 12‐year time span, and in addition, expanded sampling to other sites in Burkina Faso. Our results showed that, in contrast to earlier reports, Wolbachia is present at an extremely low prevalence in natural population of An. gambiae. From 5341 samples analysed, only 29 were positive for Wolbachia by nested PCR representing 0.54% of prevalence. No positive samples were found with regular PCR. Phylogenetic analysis of 16S rRNA gene amplicons clustered across supergroup B, with some having similarity to sequences previously found in Anopheles from Burkina Faso. However, we cannot discount the possibility that the amplicon positive samples we detected were due to environmental contamination or were false positives. Regardless, the lack of a prominent native infection in An. gambiae s.l. is encouraging for applications utilizing Wolbachia transinfected mosquitoes for malaria control. [ABSTRACT FROM AUTHOR]

Constant assessment of insecticide resistance levels is mandatory to implement adequate malaria control tools, but little information is available on the annual dynamics of resistance. We, therefore, monitored variations in resistance in Anopheles gambiae s.l. over four seasons during 2 years in two localities of Yaoundé: urban Etoa‐Meki and peri‐urban Nkolondom. Mosquitoes were collected seasonally at larval stage and reared to adults for insecticide susceptibility tests and molecular analysis of resistance mechanisms. Anopheles coluzzii was found in Etoa‐Meki and An. gambiae in Nkolondom. Low mortalities to pyrethroids were observed (permethrin <10%, deltamethrin <21%), and resistance extended to 5× and 10× diagnostic doses, revealing a marked increase compared to previous studies. A seasonal variation in resistance was observed with the highest levels within dry seasons in Etoa‐Meki and rainy seasons in Nkolondom. The 1014F kdr allele shows a high frequency (0.9), associated with overexpression of metabolic genes (Cyp6M2, Cyp6P4, Cyp9K1, Cyp6Z1, and Cyp6Z2) varying significantly seasonally. This study reveals an escalation in resistance to pyrethroids in Yaoundé's malaria vectors with seasonal variations. An adequate choice of the implementation period of punctual vector control actions according to the resistance profile will help to potentiate the desired effect and thus improve its efficiency. [ABSTRACT FROM AUTHOR]

Malaria is among the leading causes of death in Uganda, and Anopheles gambiae sensu stricto (s.s.) is the predominant vector. Although current vector control interventions have greatly reduced the malaria burden, the disease persists. New interventions are needed in order to eradicate them. Evaluation of new tools will require the availability of well‐characterized test vector populations. Juvenile An. gambiae s.s. from Kibbuye and Kayonjo‐derived populations were characterized under semi‐field and laboratory conditions, given that various vector traits, including abundance and fitness are dependent on development profiles at this life stage. Ten replicates comprising 30 first instar larvae each were profiled for various life‐history attributes (egg hatching, larval development time, larval survivorship, pupal weight and pupation rate). All parameters were similar for the two sites under laboratory conditions. However, the similarities or differences between field and laboratory development were parameter‐specific. Whereas, larval survivorship and pupal weight were similar across seasons and laboratory in colonies from both sites, in the semi‐field settings, pupation rate and larval survivorship differed between seasons in both sites. In addition, the average larval development time during the wet season was longer than that of the laboratory for both sites. Availability of mirror field sites is important for future tool evaluations. [ABSTRACT FROM AUTHOR]

Species of the genus Anopheles vary with regard to their vector capacity for Plasmodium spp., the causative agent of malaria, and their accurate identification is often required. Loop‐mediated isothermal amplification (LAMP) is a rapid, simple and low‐cost method for specific DNA amplification. Primers for LAMP assays specific for the Anopheles funestus group and Anopheles gambiae complex species as well as for the species Anopheles arabiensis, An. funestus, An. gambiae s.s/Anopheles coluzzii (major vectors) and Anopheles rivulorum (minor vector) were designed targeting specific genome or rDNA internal transcribed spacer regions. Reaction conditions (buffer composition, primer concentrations, incubation time) were evaluated and the specificities of the assays confirmed with DNA from non‐target Anopheles species. DNA release from the mosquitoes is achieved simply by heating them for 5 min in water. An aliquot of the DNA solutions is transferred to the reaction tube using disposable inoculation loops. The outcome of the LAMP amplifications after 1 h incubation at 65 °C can easily be visualized by a colour change visible to the naked eye. The assays are operable under field conditions requiring only basic equipment (portable heat block programmable at 65 and 80 °C, cooler for master mixes). [ABSTRACT FROM AUTHOR]

The microbial larvicides Bacillus thuringiensis var. israelensis (Bti) and Bacillus sphaericus (Bs) (Bacillales: Bacillaceae) are well known for their efficacy and safety in mosquito control. In order to assess their potential value in future mosquito control strategies in western Kenya, the current study tested the susceptibility of five populations of Anopheles gambiae complex mosquitoes (Diptera: Culicidae), collected from five diverse ecological sites in this area, to Bti and Bs under laboratory conditions. In each population, bioassays were conducted with eight concentrations of larvicide (Bti/Bs) in four replicates and were repeated on three separate days. Larval mortality was recorded at 24 h or 48 h after the application of larvicide and subjected to probit analysis. A total of 2400 An. gambiae complex larvae from each population were tested for their susceptibility to Bti and Bs. The mean (± standard error of the mean, SEM) lethal concentration values of Bti required to achieve 50% and 95% larval mortality (LC50 and LC95) across the five populations were 0.062  (± 0.005) mg/L and 0.797 (± 0.087) mg/L, respectively. Corresponding mean  (± SEM) values for Bs were 0.058 (± 0.005) mg/L and 0.451 (± 0.053) mg/L, respectively. Statistical analysis indicated that the five populations of An. gambiae complex mosquitoes tested were fully susceptible to Bti and Bs, and there was no significant variation in susceptibility among the tested populations. [ABSTRACT FROM AUTHOR]

New genetic control methods for mosquitoes may reduce vector species without direct effects on other species or the physical environment common with insecticides or drainage. Effects on predators and competitors could, however, be a concern as Anopheles gambiae s.l. is preyed upon in all life stages. We overview the literature and assess the strength of the ecological interactions identified. Most predators identified consume many other insect species and there is no evidence that any species preys exclusively on any anopheline mosquito. There is one predatory species with a specialisation on blood‐fed mosquitoes including An. gambiae s.l.. Evarcha culicivora is a jumping spider, known as the vampire spider, found around Lake Victoria. There is no evidence that these salticids require Anopheles mosquitoes and will readily consume blood‐fed Culex. Interspecific competition studies focus on other mosquitoes of larval habitats. Many of these take place in artificial cosms and give contrasting results to semi‐field studies. This may limit their extrapolation regarding the potential impact of reduced An. gambiae numbers. Previous mosquito control interventions are informative and identify competitive release and niche opportunism; so while the identity and relative abundance of the species present may change, the biomass available to predators may not. Genetic control methods for anopheline mosquitoes may reduce their populations without direct effects on other species or the physical environment; indirect effects on predators or competitors may be of concern.The vast majority of Anopheles gambiae predators are broad generalists and consume many other insect species; there is no indication of a tightly linked predator of adults or larvae.Interspecific competition studies indicate potential for the competitive release of other species and it is unlikely that the total mosquito biomass available to predators would alter substantially. [ABSTRACT FROM AUTHOR]

Malaria control is dependent on the use of longlasting insecticidal nets (LLINs) containing pyrethroids. A new generation of LLINs containing both pyrethroids and the synergist piperonyl butoxide (PBO) has been developed in response to increasing pyrethroid resistance in African malaria vectors, but questions remain about the performance of these nets in areas where levels of pyrethroid resistance are very high. This study was conducted in two settings in southwest Burkina Faso, Vallée du Kou 5 and Tengrela, where Anopheles gambiae s.l. (Diptera: Culicidae) mortality rates in World Health Organization (WHO) discriminating dose assays were < 14% for permethrin and < 33% for deltamethrin. When mosquitoes were pre‐exposed to PBO in WHO tube assays, mortality rates increased substantially but full susceptibility was not restored. Molecular characterization revealed high levels of kdr alleles and elevated levels of P450s previously implicated in pyrethroid resistance. In cone bioassays and experimental huts, PBO LLINs outperformed the pyrethroid‐only equivalents from the same manufacturers. Blood feeding rates were 1.6–2.2‐fold lower and mortality rates were 1.69–1.78‐fold greater in huts with PBO LLINs vs. non‐PBO LLINs. This study indicates that PBO LLINs provide greater personal and community‐level protection than standard LLINs against highly pyrethroid‐resistant mosquito populations. Pyrethroid‐treated nets are widely used to prevent malaria transmission, but resistance to these insecticides is now widespread in African malaria vectors.Nets containing the pyrethroid synergist piperonyl butoxide (PBO) are available, but the relative performance of standard vs. PBO‐impregnated nets has not been assessed in different settings.This study shows that PBO‐treated nets increase mosquito mortality and reduce blood feeding success in an area in which the mosquito population is extremely resistant to pyrethroids. [ABSTRACT FROM AUTHOR]

Abstract: The relative efficacy of a mechanical (Prokopack) collection method vs. manual aspiration in the collection of resting mosquitoes was evaluated in northern Tanzania before and after an intervention using indoor residual spraying and longlasting insecticide‐treated nets. In smoke‐free houses mosquitoes were collected from the roof and walls, but in smoky houses mosquitoes were found predominantly on the walls. Anopheles gambiae (Diptera: Culicidae) constituted 97.7% of the 312 An. gambiae complex specimens identified before but only 19.3% of the 183 identified after the intervention. A single sampling with the Prokopack collected a third of the available insects. Anopheles gambiae completed its gonotrophic development indoors, whereas Anopheles arabiensis did so outdoors. In both species gonotrophic development took 2 days. Most unfed resting An. arabiensis collected outdoors were virgins, whereas the majority of engorged insects were parous (with well‐contracted sacs). Daily survival was estimated to be 80.0%. Only 9.4% of the engorged An. arabiensis collected outdoors and 47.1% of those collected indoors had fed on humans. Using the Prokopack sampler is more efficient than manual methods for the collection of resting mosquitoes. Malaria transmission may have been affected by a change in vector composition resulting from a change in feeding, rather than reduced survival. Monitoring the proportions of members of the An. gambiae complex may provide signals of an impending breakdown in control. [ABSTRACT FROM AUTHOR]

The host preferences of the anthropophilic mosquito species in the Anopheles gambiae complex (Diptera: Culicidae) are mediated by skin bacterial volatiles. However, it is not known whether these mosquitoes respond differentially to skin bacterial volatiles from non-human host species. In this study, the responses of two malaria mosquito species in the An. gambiae complex, Anopheles gambiae s.s. (hereafter, An. gambiae) and An opheles arabiensis, with different host preferences, to volatiles released from skin bacteria were tested. Skin bacteria collected from human, cow and chicken skin significantly increased trap catches; traps containing bacteria collected from human skin caught the highest proportions of An. gambiae and An. arabiensis. Traps with bacteria of human origin caught a significantly higher proportion of An. gambiae than of An. arabiensis, whereas bacterial volatiles from the chicken attracted significantly higher numbers of An. arabiensis than of An. gambiae. Additionally, An. gambiae showed a specialized response to volatiles from four specific bacteria, whereas An. arabiensis responded equally to all species of bacteria tested. Skin bacterial volatiles may therefore play important roles in guiding mosquitoes with different host preferences. The identification of these bacterial volatiles can contribute to the development of an odour blend that attracts mosquitoes with different host preferences. [ABSTRACT FROM AUTHOR]

Insecticide resistance and growing public concern over the safety and environmental impacts of some conventional insecticides have resulted in the need to discover alternative control tools. Naturally occurring botanically-based compounds are of increased interest to aid in the management of mosquitoes. Susceptible strains of Aedes aegypti (Linnaeus) ( Diptera: Culicidae) and Anopheles gambiae (Meigen) ( Diptera: Culicidae) were treated with permethrin, a common type- I synthetic pyrethroid, using a discriminate dose that resulted in less than 50% mortality. Piperonyl butoxide ( PBO) and 35 essential oils were co-delivered with permethrin at two doses (2 and 10 µg) to determine if they could enhance the 1-h knockdown and the 24-h mortality of permethrin. Several of the tested essential oils enhanced the efficacy of permethrin equally and more effectively than piperonyl butoxide PBO, which is the commercial standard to synergize chemical insecticide like pyrethroids. PBO had a strikingly negative effect on the 1-h knockdown of permethrin against Ae. aegypti, which was not observed in An. gambiae. Botanical essential oils have the capability of increasing the efficacy of permethrin allowing for a natural alternative to classic chemical synergists, like PBO. [ABSTRACT FROM AUTHOR]

Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is a vector for Plasmodium, the causative agent of malaria. Current control strategies to reduce the impact of malaria focus on reducing the frequency of mosquito attacks on humans, thereby decreasing Plasmodium transmission. A need for new repellents effective against Anopheles mosquitoes has arisen because of changes in vector behaviour as a result of control strategies and concern over the health impacts of current repellents. The response of A. gambiae to potential repellents was investigated through an electroantennogram screen and the most promising of these candidates (1-allyloxy-4-propoxybenzene, 3c{ 3,6}) chosen for behavioural testing. An assay to evaluate the blood-host seeking behaviour of A. gambiae towards a simulated host protected with this repellent was then performed. The compound 3c{ 3,6} was shown to be an effective repellent, causing mosquitoes to reduce their contact with a simulated blood-host and probe less at the host odour. Thus, 3c{ 3,6} may be an effective repellent for the control of A. gambiae. [ABSTRACT FROM AUTHOR]

Long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) are the cornerstones of malaria vector control. However, the effectiveness of these control tools depends on vector ecology and behaviour, which also largely determine the efficacy of certain Anopheles mosquitoes (Diptera: Culicidae) as vectors. Malaria vectors in sub-Saharan Africa are primarily species of the Anopheles gambiae complex, which present intraspecific differences in behaviour that affect how they respond to vector control tools. The focus of this study is the change in species composition in the An. gambiae complex after the implementation of LLINs in Dielmo, Senegal. The main findings referred to dramatic decreases in the proportions of Anopheles coluzzii and An. gambiae after the introduction of LLINs, and an increase in the proportion of Anopheles arabiensis. Two years after LLINs were first introduced, An. arabiensis remained the most prevalent species and An. gambiae had begun to rebound. This indicated a need to develop additional vector control tools that can target the full range of malaria vectors. [ABSTRACT FROM AUTHOR]

Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is the major Afro-tropical vector of malaria. Novel strategies proposed for the elimination and eradication of this mosquito vector are based on the use of genetic approaches, such as the sterile insect technique ( SIT). These approaches rely on the ability of released males to mate with wild females, and depend on the application of effective protocols to assess the swarming and mating behaviours of laboratory-reared insects prior to their release. The present study evaluated whether large semi-field enclosures can be utilized to study the ability of males from a laboratory colony to respond to natural environmental stimuli and initiate normal mating behaviour. Laboratory-reared males exhibited spatiotemporally consistent swarming behaviour within the study enclosures. Swarm initiation, peak and termination time closely tracked sunset. Comparable insemination rates were observed in females captured in copula in the semi-field cages relative to females in small laboratory cages. Oviposition rates after blood feeding were also similar to those observed in laboratory settings. The data suggest that outdoor enclosures are suitable for studying swarming and mating in laboratory-bred males in field-like settings, providing an important reference for future studies aimed at assessing the comparative mating ability of strains for SIT and other vector control strategies. [ABSTRACT FROM AUTHOR]

A large-scale mesocosm was constructed and tested for its effectiveness for use in experiments on behaviour, reproduction and adult survivorship in the Afrotropical malaria vector Anopheles gambiae s.s. Giles ( Diptera: Culicidae) in temperate climates. The large space (82.69 m3) allowed for semi-natural experiments that increased demand on a mosquito's energetic reserves in an environment of widely distributed resources. A one-piece prefabricated enclosure, made with white netting and vinyl, prevented the ingress of predators and the egress of mosquitoes. Daylight and white materials prompted the mosquitoes to seclude themselves in restricted daytime resting sites and allowed the easy collection of dead bodies so that daily mortality could be assessed accurately using a method that accounts for the loss of a proportion of bodies. Here, daily, age-dependent mortality rates of males and females were estimated using Bayesian Markov chain Monte Carlo simulation. In overnight experiments, mosquitoes successfully located plants and took sugar meals. A 3-week survival trial with a single cohort demonstrated successful mating, blood feeding, oviposition and long life. The relatively low cost of the mesocosm and the performance of the mosquitoes in it make it a viable option for any behavioural or ecological study of tropical mosquitoes in which space and seasonal cold are constraining factors. [ABSTRACT FROM AUTHOR]

Comparisons were undertaken to investigate cost-effective methods of implementing the enzyme-linked immunosorbent assay ( ELISA) for sporozoite determination in anophelines when large numbers require processing. Comparisons between ELISA plate reader and visual assessments were performed with Anopheles funestus and Anopheles gambiae s.l. ( Diptera: Culicidae), as were comparisons between whole-body mosquito samples, heads and thoraces, and abdomens alone. Rates obtained from pools of five or 10 mosquitoes were compared with those for individual mosquitoes, as were rates obtained using different sampling methods. A total of 41 792 An. funestus and 9431 An. gambiae s.l. collected in light traps, and 22 323 An. funestus and 6860 An. gambiae s.l . from exit collections were analysed. Visual assessments gave results similar to those of machine readings. Sporozoite rates were similar in both species, as were rates by collection method. The use of whole mosquitoes increased estimates of infection rate by 0.6%. Pool size did not affect infection rates of An. gambiae s.l., but rates were higher among individually tested An. funestus than among those tested in pools. For large-scale surveys, the use of whole mosquitoes in pools of 10 mosquitoes, with correction for overestimation, and the noting of results according to a simple three-stage visual assessment of positivity is the most cost-effective approach and is sufficient to obtain reliable data for comparative purposes. [ABSTRACT FROM AUTHOR]

The emergence of insecticide resistance in Anopheles ( Diptera: Culicidae) mosquitoes has great implications for malaria control in Nigeria. This study aimed to determine the dynamics of insecticide susceptibility levels and the frequency of knock-down resistance ( kdr) mutations ( L1014F) in wild Anopheles coluzzii Coetzee & Wilkerson sp. n. and Anopheles gambiae Giles from the Ojoo and Bodija areas of Ibadan, in southwest Nigeria. Insecticide susceptibility to pyrethroids, organophosphates, carbamates and organochlorines was assessed using World Health Organization ( WHO) bioassays. A subset of the mosquitoes exposed to pyrethroids and DDT was used for species and molecular form identification; kdr genotyping was determined using the Taq Man real-time polymerase chain reaction assay. The mosquitoes were resistant to pyrethroids and DDT but completely susceptible to organophosphates and carbamates. Bodija samples ( n = 186) consisted of An. gambiae (91.4%) and An. coluzzii (8.1%) and included one An. coluzzii/ An. gambiae hybrid specimen. All mosquitoes screened in Ojoo ( n = 26) were An. gambiae. The 1014F kdr mutation was detected at frequencies of 24.5 and 5.8% in Bodija and Ojoo, respectively. No correlation was observed between kdr genotypes and resistance phenotypes. The results indicate that metabolic resistance probably plays an important role in the development of resistance and highlight the need to implement insecticide resistance management strategies. [ABSTRACT FROM AUTHOR]

Anopheles gambiae s.l. ( Diptera: Culicidae) in Muleba, Tanzania has developed high levels of resistance to most insecticides currently advocated for malaria control. The kdr mutation has almost reached fixation in An. gambiae s.s. in Muleba. This change has the potential to jeopardize malaria control interventions carried out in the region. Trends in insecticide resistance were monitored in two intervention villages using World Health Organization ( WHO) susceptibility test kits. Additional mechanisms contributing to observed phenotypic resistance were investigated using Centers for Disease Control ( CDC) bottle bioassays with piperonylbutoxide ( PBO) and S,S,S-tributyl phosphorotrithioate ( DEF) synergists. Resistance genotyping for kdr and Ace-1 alleles was conducted using quantitative polymerase chain reaction ( qPCR). In both study villages, high phenotypic resistance to several pyrethroids and DDT was observed, with mortality in the range of 12-23%. There was a sharp decrease in mortality in An. gambiae s.l. exposed to bendiocarb (carbamate) from 84% in November 2011 to 31% in December 2012 after two rounds of bendiocarb-based indoor residual spraying ( IRS). Anopheles gambiae s.l. remained susceptible to pirimiphos-methyl (organophosphate). Bendiocarb-based IRS did not lead to the reversion of pyrethroid resistance. There was no evidence for selection for Ace-1 resistance alleles. The need to investigate the operational impact of the observed resistance selection on the effectiveness of longlasting insecticidal nets and IRS for malaria control is urgent. [ABSTRACT FROM AUTHOR]

Mosquito repellents are used around the globe to protect against nuisance biting and disease-transmitting mosquitoes. Recently, there has been renewed interest in the development of repellents as tools to control the transmission of mosquito-borne diseases. We present a new bioassay for the accurate assessment of candidate repellent compounds, using a synthetic odour that mimics the odour blend released by human skin. Using DEET ( N, N-diethyl-meta-toluamide) and PMD ( p-menthane-3,8-diol) as reference compounds, nine candidate repellents were tested, of which five showed significant repellency to the malaria mosquito Anopheles gambiae sensu stricto (Diptera: Culicidae). These included: 2-nonanone; 6-methyl-5-hepten-2-one; linalool; δ-decalactone, and δ-undecalactone. The lactones were also tested on the yellow fever mosquito Aedes aegypti ( Stegomyia aegypti) (Diptera: Culicidae), against which they showed similar degrees of repellency. We conclude that the lactones are highly promising repellents, particularly because these compounds are pleasant-smelling, natural products that are also present in human food sources. [ABSTRACT FROM AUTHOR]

The development of insecticide resistance is a threat to the control of malaria in Africa. We report the findings of a national survey carried out in Tanzania in 2011 to monitor the susceptibility of malaria vectors to pyrethroid, organophosphate, carbamate and DDT insecticides, and compare these findings with those identified in 2004 and 2010. Standard World Health Organization ( WHO) methods were used to detect knock-down and mortality rates in wild female Anopheles gambiae s.l. (Diptera: Culicidae) collected from 14 sentinel districts. Diagnostic doses of the pyrethroids deltamethrin, lambdacyhalothrin and permethrin, the carbamate propoxur, the organophosphate fenitrothion and the organochlorine DDT were used. Anopheles gambiae s.l. was resistant to permethrin in Muleba, where a mortality rate of 11% [95% confidence interval (CI) 6-19%] was recorded, Muheza (mortality rate of 75%, 95% CI 66-83%), Moshi and Arumeru (mortality rates of 74% in both). Similarly, resistance was reported to lambdacyhalothrin in Muleba, Muheza, Moshi and Arumeru (mortality rates of 31-82%), and to deltamethrin in Muleba, Moshi and Muheza (mortality rates of 28-75%). Resistance to DDT was reported in Muleba. No resistance to the carbamate propoxur or the organophosphate fenitrothion was observed. Anopheles gambiae s.l. is becoming resistant to pyrethoids and DDT in several parts of Tanzania. This has coincided with the scaling up of vector control measures. Resistance may impair the effectiveness of these interventions and therefore demands close monitoring and the adoption of a resistance management strategy. [ABSTRACT FROM AUTHOR]

Anopheles moucheti Evans (Diptera: Culicidae) is a major vector of malaria in forested areas of Central Africa. However, few genetic tools are available for this species. The present study represents the first attempt to characterize chromosomes in An. moucheti females collected in Cameroon. Ovarian nurse cells contained polytene chromosomes, which were suitable for standard cytogenetic applications. The presence of three polymorphic chromosomal inversions in An. moucheti was revealed. Two of these inversions were located on the 2R chromosome arm. The homology between the 2R chromosome arms of An. moucheti and Anopheles gambiae Giles was established by fluorescent in situ hybridization of six An. gambiae genic sequences. Mapping of the probes on chromosomes of An. moucheti detected substantial gene order reshuffling between the two species. The presence of polytene chromosomes and polymorphic inversions in An. moucheti provides a new basis for further population genetic, taxonomic and ecological studies of this neglected malaria vector. [ABSTRACT FROM AUTHOR]

Previously, essential oil of Conyza newii (Asterale: Asteracea, Oliv. & Hiern) growing in the northern part of West Pokot (35°E, 1°N) of Kenya was shown to be highly repellent [ RD50 = 8.9 × 10−5 mg/cm2, 95% confidence interval ( CL)] to Anopheles gambiae s.s. Fumigant toxicity of the oil to the mosquito was also demonstrated. The major constituents of the oil were found to be monoterpenoids, including ( S)-(-)-perillyl alcohol, ( S)-(-)-perillaldehyde, geraniol, ( R)-(+)-limonene, trans-β-ocimene and 1,8-cineol. In this study, the chemical composition and repellency of essential oils of the plant seedlings collected from West Pokot (35°E, 1°N) and propagated in seven different geographical regions of Kenya [West Pokot (35°E, 1°N), Kilome (37°E, 1°S), Naivasha (36°E, 0°), Webuye (34°E, 1°N), Nyakach (34°E, 0°), Kericho (35°E, 0°) and Nairobi (36°E, 1°S)] were compared. There were significant variations ( P < 0.01, 95% CL) in the relative proportions of the six constituents and this was reflected in the repellency of the essential oils ( P < 0.01, 95% CL). Higher repellency of the oil was associated with greater proportions of ( S)-(-) perillyl alcohol, ( S)-(-)-perillaldehyde and geraniol, and lower repellency was associated with an increased proportion of ( R)-(+)-limonene. The results suggest significant epigenetic (chemotypic) variations in the repellency and composition of C. newii essential oils growing in different regions of Kenya. [ABSTRACT FROM AUTHOR]

A strategy to decrease the vector competence of Anopheles gambiae sensu stricto (Diptera: Culicidae), the most efficient malaria vector in Africa, may consist of exploiting the genes involved in zoophily. Crossing and backcrossing experiments were performed between An. gambiae s.s. and the zoophilic sibling species Anopheles quadriannulatus. Mosquito strains were tested in a dual-choice olfactometer to investigate their responses to cow odour. Totals of 12% of An. gambiae s.s. and 59% of An. quadriannulatus selected the port with the cow odour. Crosses and backcrosses did not show a significant preference for the cow-baited port. The results indicated that anthropophilic behaviour in An. gambiae s.s. is a dominant or partially dominant trait, which, in conjunction with the unstable zoophilic behaviour observed in An. quadriannulatus, poses a serious obstacle to plans to decrease vector competence by modifying the anthropophilic trait. [ABSTRACT FROM AUTHOR]

A DNA-DNA hybridization method, reverse dot blot analysis (RDBA), was used to identify Anopheles gambiae s.s. and Anopheles arabiensis (Diptera: Culicidae) hosts. Of 299 blood-fed and semi-gravid An. gambiae s.l. collected from Kisian, Kenya, 244 individuals were identifiable to species; of these, 69.5% were An. arabiensis and 29.5% were An. gambiae s.s. Host identifications with RDBA were comparable with those of conventional polymerase chain reaction (PCR) followed by direct sequencing of amplicons of the vertebrate mitochondrial cytochrome b gene. Of the 174 amplicon-producing samples used to compare these two methods, 147 were identifiable by direct sequencing and 139 of these were identifiable by RDBA. Anopheles arabiensis bloodmeals were mostly (94.6%) bovine in origin, whereas An. gambiae s.s. fed upon humans more than 91.8% of the time. Tests by RDBA detected that two of 112 An. arabiensis contained blood from more than one host species, whereas PCR and direct sequencing did not. Recent use of insecticide-treated bednets in Kisian is likely to have caused the shift in the dominant vector species from An. gambiae s.s. to An. arabiensis. Reverse dot blot analysis provides an opportunity to study changes in host-feeding by members of the An. gambiae complex in response to the broadening distribution of vector control measures targeting host-selection behaviours. [ABSTRACT FROM AUTHOR]

The molecular form composition of Anopheles gambiae Giles s.s. (Diptera: Culicidae) mating swarms and the associated mating pairs ( copulae) were investigated during two rainy seasons (July to October, 2005 and July to November, 2006) in the villages of Soumousso and Vallée du Kou (VK7). Although the habitats of these villages differ markedly, sympatric populations of M and S molecular forms of An. gambiae s.s. occur in both places periodically. The main aim was to assess the degree to which these molecular forms mate assortatively. In Soumousso, a wooded savannah habitat, the majority of swarm samples consisted of only S-form males (21/28), although a few M-form males were found in mixed M- and S-form swarms. In VK7, a rice growing area, the majority of swarm samples consisted of only M-form males (38/62), until October and November 2006, when there were nearly as many mixed-form as single-form swarms. Overall, ∼60% of M- and S-form swarms were temporally or spatially segregated; the two forms were effectively prevented from encountering each other. Of the remaining 40% of swarms, however, only about half were single-form and the rest were mixed-form. Of the 33 copulae collected from mixed-form swarms, only four were mixed-form pairs, significantly fewer than expected by random pairing between forms ( χ2 = 10.34, d.f. = 2, P < 0.01). Finally, all specimens of inseminated females were of the same form as the sperm contained within their spermatheca ( n = 91), even for the four mixed-form copulae. These findings indicate that assortative mating occurs within mixed-form swarms, mediated most probably by close-range mate recognition cues. [ABSTRACT FROM AUTHOR]

Insecticide resistance in the malaria vector Anopheles gambiae s.l. (Diptera: Culicidae) threatens insecticide-based control efforts, necessitating regular monitoring. We assessed resistance in field-collected An. gambiae s.l. from Jinja, Uganda using World Health Organization (WHO) biosassays. Only An. gambiae s.s. and An. arabiensis (≅70%) were present. Female An. gambiae exhibited extremely high pyrethroid resistance (permethrin LT50 > 2 h; deltamethrin LT50 > 5 h). Female An. arabiensis were resistant to permethrin and exhibited reduced susceptibility to deltamethrin. However, while An. gambiae were DDT resistant, An. arabiensis were fully susceptible. Both species were fully susceptible to bendiocarb and fenitrothion. Kdr 1014S has increased rapidly in the Jinja population of An. gambiae s.s. and now approaches fixation (≅95%), consistent with insecticide-mediated selection, but is currently at a low frequency in An. arabiensis (0.07%). Kdr 1014F was also at a low frequency in An. gambiae. These frequencies preclude adequately-powered tests for an association with phenotypic resistance. PBO synergist bioassays resulted in near complete recovery of pyrethroid susceptibility suggesting involvement of CYP450s in resistance. A small number (0.22%) of An. gambiae s.s. × An. arabiensis hybrids were found, suggesting the possibility of introgression of resistance alleles between species. The high levels of pyrethroid resistance encountered in Jinja threaten to reduce the efficacy of vector control programmes which rely on pyrethroid-impregnated bednets or indoor spraying of pyrethroids. [ABSTRACT FROM AUTHOR]

Field and laboratory investigations revealed phenotypic, target site and metabolic resistance to permethrin in an Anopheles gambiae s.s. (Diptera: Culicidae) population in Bungoma District, a region in western Kenya in which malaria is endemic and rates of ownership of insecticide-treated bednets are high. The sensitivity of individual An. gambiae s.l. females as indicated in assays using World Health Organization (WHO) test kits demonstrated reduced mortality in response to permethrin, deltamethrin and bendiocarb. Estimated time to knock-down of 50% (KDT50) of the test population in Centers for Disease Control (CDC) bottle bioassays was significantly lengthened for the three insecticides compared with that in a susceptible control strain. Anopheles arabiensis from all three sites showed higher mortality to all three insecticides in the WHO susceptibility assays compared with the CDC bottle assays, in which they showed less sensitivity and longer KDT50 than the reference strain for permethrin and deltamethrin. Microplate assays revealed elevated activity of β-esterases and oxidases, but not glutathione-S-transferase, in An. gambiae s.s. survivors exposed to permethrin in bottle bioassays compared with knocked down and unexposed individuals. No An. arabiensis showed elevated enzyme activity. The 1014S kdr allele was fixed in the Bungoma An. gambiae s.s. population and absent from An. arabiensis, whereas the 1014F kdr allele was absent from all samples of both species. Insecticide resistance could compromise vector control in Bungoma and could spread to other areas as coverage with longlasting insecticide-treated bednets increases. [ABSTRACT FROM AUTHOR]

In West Africa, M and S molecular forms of Anopheles gambiae sensu stricto (Diptera: Culicidae) Giles, frequently occur together, although with different population bionomics. The S form typically breeds in rain-dependant water collections and is present during the rainy season only whereas the M form can thrive all year long in areas with permanent breeding opportunities. In the present study, we explored physiological and developmental trade-offs at play in laboratory colonies and field populations of the M and S forms that originated from an area of sympatry in Burkina Faso, where M and S larvae exhibit such habitat segregation. In the laboratory, larvae of the M form developed slower than the S form (mean values 9.51 and 8.85 days, respectively, Wilcoxon's test, P < 0.001). Although wing length and dry weight at emergence showed large variations, M females were on average 8% heavier than S females of similar wing length. Higher nutritional reserves (proteins and lipids) in teneral adults explained part of this weight difference, reflecting a better ability of the M form to garner resources at the larval stage. Furthermore, a higher rate of ovarian maturation was observed in the M form after a single bloodmeal. The relevance of these findings for parasite transmission is discussed. [ABSTRACT FROM AUTHOR]

Malaria transmission was monitored in two villages in the Sahel zone of Niger over 4 years. During this period, a nationwide vector control programme was carried out in which insecticide-treated bednets were distributed free to mothers of children aged <5 years. Anopheles gambiae and Anopheles arabiensis (Diptera: Culicidae) were found to be the major malaria vectors. The dynamics of An. gambiae s.l. did not vary dramatically over the study period although the proportion of female mosquitoes found resting indoors decreased in both villages and, in one village, the parity rate and sporozoite index were significantly reduced after bednet distribution. By contrast with An. gambiae, the dynamics of Anopheles funestus altered greatly after the bednet distribution period, when adult density, endophagous rate and sporozoite rates decreased dramatically. Our observations highlight the importance of quantifying and monitoring the dynamics and infections of malaria vectors during large-scale vector control interventions. [ABSTRACT FROM AUTHOR]

Mortality rates, determined by dissection, of predominantly M form female Anopheles gambiae (Diptera: Culicidae) were estimated. Mosquitoes were collected in tent traps and light traps in an irrigation project village in Ghana in June and July 2010, when much of the area was flooded. Both M and S form larvae were collected from rice fields (74 of 80 specimens were M form). Adults were collected in equal proportions from the two traps (90 of 107 specimens from the light trap and 106 of 116 specimens from the tent trap were M form). During the study, collection numbers rose from 105 to 972 per night. A total of 1787 of the 15 431 An. gambiae collected were dissected. Of these, 953 (53%) were found to have taken their first bloodmeal, either as virgins or following mating. The age profiles of mosquitoes collected alive and dead, respectively, were similar. Eighteen of 2933 (0.61 ± 0.49%) specimens were found to be positive for sporozoites in an enzyme-linked immunosorbent assay (ELISA). Lagged cross correlations among the different age groups implied that the mosquitoes fed on days 2 and 4 following emergence prior to oviposition and every 2.65 ± 0.17 days thereafter. The best model to describe the observed population patterns implied a daily mortality of 84%. The results are discussed in relation to possible mosquito control measures for the village. [ABSTRACT FROM AUTHOR]

The M and S molecular forms of the African malaria vector Anopheles gambiae (Diptera: Culicidae) are morphologically identical incipient species in which reproductive isolation is incomplete, enabling low-level gene flow between forms. In an attempt to find differences between the M and S forms, sequence variation was studied at loci along the X chromosome in adult female An. gambiae from Angola. A high proportion of M form specimens from Angola (79% of the 456 X chromosomes sampled) were found to contain a 16-bp insertion in intron 4 of the X-linked GPRCCK1 locus, relative to the AgamP3 release of the An. gambiae PEST genome sequence. The insertion was in Hardy-Weinberg equilibrium in Angolan M form populations. The same insertion was found in all S form specimens examined, regardless of where in Africa they were sampled, but was absent from a sample of M form specimens collected in Ghana, Bioko and Mali. In M form specimens from Angola, there was an association between alleles at the GPRCCK1 locus and those at a microsatellite locus, AGXH678, close to the centromere of the X chromosome, with significant linkage disequilibrium between loci separated by 0.472 Mbp ( P < 0.033). We show that the insertion results from introgression from the S form into the M form, rather than from the retention of an ancestral character. Gene flow from the S to M form could allow genes of adaptive value to be transferred, including those conferring insecticide resistance and others influencing ecology and behaviour, and thus malaria transmission and control. We discuss factors that may have led to this introgression event. [ABSTRACT FROM AUTHOR]

In West Africa, lineage splitting between the M and S molecular forms of the major Afro-tropical malaria mosquito, Anopheles gambiae (Diptera: Culicidae), is thought to be driven by ecological divergence, occurring mainly at the larval stage. Here, we present evidence for habitat segregation between the two molecular forms in and around irrigated rice fields located within the humid savannahs of western Burkina Faso. Longitudinal sampling of adult mosquitoes emerging from a range of breeding sites distributed along a transect extending from the heart of the rice field area into the surrounding savannah was conducted from June to November 2009. Analysis revealed that the two molecular forms and their sibling species Anopheles arabiensis are not randomly distributed in the area. A major ecological gradient was extracted in relation to the perimeter of the rice fields. The M form was associated with larger breeding sites mostly consisting of rice paddies, whereas the S form and An. arabiensis were found to depend upon temporary, rain-filled breeding sites. These results support hypotheses about larval habitat segregation and confirm the suggestion that the forms have different larval habitat requirements. Segregation appears to be clearly linked to anthropogenic permanent habitats and the community structure they support. [ABSTRACT FROM AUTHOR]

Host-seeking behaviour of the anthropophilic malaria vector Anopheles gambiae sensu stricto (Diptera: Culicidae) is mediated predominantly by olfactory cues. Several hundreds of odour components have been identified from human emanations, but only a few have been proven to act as attractants or synergists in the host-seeking behaviour of female An. gambiae. In previous work, aromatics, alcohols and ketones in human odours were found to elicit electrophysiological activity in antennal olfactory neurons of female An. gambiae. However, the behavioural effects of these compounds have not been investigated. In this study, behavioural responses of female An. gambiae to components of human breath, urine and sweat at a series of concentrations, or a single concentration in the case of acetone, were examined in combination with ammonia and L-lactic acid in a dual-choice olfactometer. The results showed that at specific concentrations 4-ethylphenol, indole, 3-methyl-1-butanol and two ketones inhibited the attractive effect of a mixture of ammonia and lactic acid. Acetone on its own was not attractive; however, when combined with lactic acid, the binary mixture was attractive. When combined with ammonia, acetone inhibited the attractiveness exerted by ammonia alone. Dodecanol and dimethyldisulphide did not affect the attraction exerted by ammonia and lactic acid at any of the concentrations tested. By contrast, a human-specific armpit odour, 7-octenoic acid, augmented the attraction exerted by the combination of ammonia and lactic acid at a specific dosage. [ABSTRACT FROM AUTHOR]

Benin has embraced World Health Organization-recommended preventive strategies to control malaria. Its National Malaria Control Programme is implementing and/or coordinating various actions and conducting evaluation trials of mosquito control strategies. Mosquito control is based on the use of insecticide-treated nets and indoor residual spraying, but the efficacy of these strategies to control malaria vectors is endangered by insecticide resistance. Here, we present the results of a nationwide survey on the status of insecticide susceptibility and resistance in Anopheles gambiae s.l. (Diptera: Culicidae) carried out in Benin in 2006-2007 (i.e. before extensive vector control was undertaken). Overall, our study showed that the S molecular form of An. gambiae s.s. predominates and is widely distributed across the country, whereas the frequency of the M form shows a strong decline with increasing latitude. Susceptibility to DDT, permethrin, carbosulfan and chlorpyrifos-methyl was assessed; individual mosquitoes were identified for species and molecular forms, and genotyped for the kdr and ace-1 loci. Full susceptibility to chlorpyrifos-methyl was recorded and very few samples displayed resistance to carbosulfan. High resistance levels to permethrin were detected in most samples and almost all samples displayed resistance to DDT. The kdr-Leu-Phe mutation was present in all localities and in both molecular forms of An. gambiae s.s. Furthermore, the ace-1 R mutation was predominant in the S form, but absent from the M form. By contrast, no target modification was observed in Anopheles arabiensis. Resistance in the An. gambiae S molecular form in this study seemed to be associated with agricultural practices. Our study showed important geographic variations which must be taken into account in the vector control strategies that will be applied in different regions of Benin. It also emphasizes the need to regularly monitor insecticide resistance across the country and to adapt measures to manage resistance. [ABSTRACT FROM AUTHOR]

The global rate of heavy metal pollution is rapidly increasing in various habitats. Anopheles malaria vector species (Diptera: Culicidae) appear to tolerate many aquatic habitats with metal pollutants, despite their normal proclivity for ‘clean’ water (i.e. low levels of organic matter). Investigations were conducted to establish whether there are biological costs for tolerance to heavy metals in Anopheles gambiae Giles sensu stricto and to assess the potential impact of heavy metal pollution on mosquito ecology. Anopheles gambiae s.s. were selected for cadmium, copper or lead tolerance through chronic exposure of immature stages to solutions of the metals for three successive generations. Biological costs were assessed in the fourth generation by horizontal life table analysis. Tolerance in larvae to cadmium (as cadmium chloride, CdCl2), copper [as copper II nitrate hydrate, Cu(NO3)2 2.5 H2O] and lead [as lead II nitrate, Pb(NO3)2], monitored by changes in LC50 concentrations of the metals, changed from 6.07 µg/L, 12.42 µg/L and 493.32 µg/L to 4.45 µg/L, 25.02 µg/L and 516.69 µg/L, respectively, after three generations of exposure. The metal-selected strains had a significantly lower magnitude of egg viability, larval and pupal survivorship, adult emergence, fecundity and net reproductive rate than the control strain. The population doubling times were significantly longer and the instantaneous birth rates lower in most metal-selected strains relative to the control strain. Our results suggest that although An. gambiae s.s. displays the potential to develop tolerance to heavy metals, particularly copper, this may occur at a significant biological cost, which can adversely affect its ecological fitness. [ABSTRACT FROM AUTHOR]

This study provides data on malaria vector species composition and insecticide susceptibility status from three localities in Guinea Conakry. A total of 497 mosquitoes were collected resting indoors and morphologically identified as belonging to the Anopheles gambiae complex. The majority of these were An. gambiae s.s. (99.6%), but a small percentage (0.4%) were identified as Anopheles arabiensis. Thirty-four Anopheles funestus s.s. were also collected. The molecular S form of An. gambiae s.s. was predominant over the M form in Siguiri (95%) and Boffa (97.4%), whereas at Mt Nimba the M form was more abundant (61.4%) than the S form (38.1%). One hybrid M/S specimen was recorded from Mt Nimba. Siguiri populations showed high levels of resistance to DDT, dieldrin and bendiocarb. Anopheles gambiae from Boffa were largely susceptible to the insecticides tested. At Mt Nimba, resistance to DDT and bendicocarb was detected. Biochemical enzyme analysis showed that an altered acetylcholinesterase is operating in the field at low levels. The frequency of the 1014F kdr allele in the An. gambiae S form was 0.24 at Siguiri and 0.14 at Mt Nimba. A single RR specimen was found in the M form. The heterogeneity in species composition and resistance profiles between sites requires vector control interventions to be tailored to each site based on the data collected from ongoing monitoring and surveillance. [ABSTRACT FROM AUTHOR]

Anopheles gambiae Giles s.s. and Anopheles arabiensis Patton (Diptera: Culicidae) are major vectors of malaria in Nigeria. We used 1115 bp of the mitochondrial COI gene to assess their population genetic structures based on samples from across Nigeria ( n = 199). The mtDNA neighbour-joining tree, based on FST estimates, separated An. gambiae M and S forms, except that samples of An. gambiae M from Calabar clustered with all the An. gambiae S form. Anopheles arabiensis and An. gambiae could be combined into a single star-shaped, parsimonious haplotype network, and shared three haplotypes. Haplotype diversity values were high in An. arabiensis and An. gambiae S, and intermediate in An. gambiae M; all nucleotide diversities were relatively low. Taken together, patterns of haplotype diversity, the star-like genealogy of haplotypes, five of seven significant neutrality tests, and the violation of the isolation-by-distance model indicate population expansion in An. arabiensis and An. gambiae S, but the signal was weak in An. gambiae M. Selection is supported as an important factor shaping genetic structure in An. gambiae in Nigeria. There were two geographical subdivisions in An. arabiensis: one included all southern localities and all but two central localities; the other included all northern and two central localities. Re-analysing an earlier microsatellite dataset of An. arabiensis using a Bayesian method determined that there were two distinctive clusters, northern and southern, that were fairly congruent with the mtDNA subdivisions. There was a trend towards decreasing genetic diversity in An. arabiensis from the northern savannah to the southern rainforest that corroborated previous data from microsatellites and polytene chromosomes. [ABSTRACT FROM AUTHOR]

Anopheles gambiae Giles s.s. (Diptera: Culicidae) is known to feed on plant sugars, but this is the first experimental study to consider whether it discriminates between plant species. Thirteen perennial plant species were selected on the basis of their local availability within the vicinity of human dwellings and larval habitats of An. gambiae s.s. in western Kenya. Groups of 100 or 200 mosquitoes were released into cages either with a cutting of one plant type at a time (single-plant assay) or with cuttings of all 13 plants simultaneously (choice assay), respectively, and left overnight. In the choice assay, direct observations of the percentages of mosquitoes perching or feeding on each plant were recorded over four 1-h periods each night. For both types of assay, mosquitoes were recaptured and the percentage that had fed on plants was assessed by testing them individually for the presence of fructose. To identify which plants the choice-assay mosquitoes had fed on, gas chromatography (GC) profiles of samples of mosquito homogenates were compared with GC profiles of extracts from relevant parts of each plant. Four of the plants that were observed to have been fed on most frequently in the choice assay ( Parthenium hysterophorus L., Tecoma stans L., Ricinus communis L., and Senna didymobotrya Fresen) were also shown to have been ingested most often by mosquitoes in both types of assay, suggesting that An. gambiae is differentially responsive to this range of plants, regardless of whether the plants were presented singly or mixed together. Significantly more females than males fed on plants, with the exception of P. hysterophorus L., one of the plants most frequently fed on. For most plant species (ten of 13), GC profiles indicated that An. gambiae obtained sugars primarily from flowers. The exceptions were P. hysterophorus L., Lantana camara L. and R. communis L., on which An. gambiae fed more often from leaves and stems than from flowers. [ABSTRACT FROM AUTHOR]

Differences between human individuals in their attractiveness to female mosquitoes have been reported repeatedly, but the underlying mechanisms are not well understood. Skin emanations from 27 human individuals, collected on glass marbles, were tested against ammonia in a dual-choice olfactometer to establish their degrees of attractiveness to anthropophilic Anopheles gambiae s.s. Giles (Diptera: Culicidae) mosquitoes. Ammonia was used as a standard odour source because of its proven attractiveness to An. gambiae s.s. Skin emanations from most volunteers attracted significantly more mosquitoes than ammonia. There were clear differences in the attractiveness of skin emanations from different volunteers relative to that of ammonia, as well as in the strength of the trap entry response. Consistent differences were observed when emanations from the three most and the three least attractive volunteers were tested pairwise. No gender or age effect was found for relative attractiveness or trap entry response. Emanations from volunteers with higher behavioural attractiveness elicited higher electroantennogram response amplitudes in two pairs, but in a third pair a higher electroantennogram response was found for the less attractive volunteer. These results confirm that odour contributes to the differences in attractiveness of humans to mosquitoes. [ABSTRACT FROM AUTHOR]

Little is known about the sugar-feeding behaviour of equatorial Africa's principal vector of malaria, Anopheles gambiae Giles (Diptera: Culicidae). It is suspected to feed on plant sugar infrequently, but possibly the timing depends on environmental circumstances, and males may differ markedly from females. These points of uncertainty were clarified in the laboratory, by monitoring both diel and longterm sugar-feeding activity in both sexes. Males fed on sugar in a nocturnal diel rhythm closely approximating non-specific flight activity. Female diel sugar-feeding patterns resembled published rhythms and cycles of host seeking. Males sugar fed nightly at an average frequency of about twice per night, sustained over 17 days. This was substantially higher than the sugar-feeding frequency of females that were allowed both blood and oviposition sites every night: they averaged about one sugar feed in every 4 nights. These females fed on sugar between gonotrophic cycles, after eggs were mature but before the next bloodmeal. They did not sugar feed during the 2 days after blood feeding, while blood was being digested and the eggs developed. A slight delay in the availability of either the oviposition site or blood led to an increase in female sugar-feeding frequency: they averaged more than once per night until the delayed resource was made available. These observations support the conclusion that sugar feeding is a normal part of the biology of both sexes of An. gambiae. [ABSTRACT FROM AUTHOR]

Anopheles gambiae Giles s.s. (Diptera: Culicidae) is one of the principal vectors of malaria in the Ashanti region of central Ghana. High levels of resistance to dieldrin were recorded in a wild-caught sample from Obuasi (south of Kumasi) as well as a laboratory colony established using material from the wild population. Cytogenetic analysis of wild-caught and laboratory samples revealed chromosomal polymorphism for inversions 2La and 2Rb. Although inversion 2La has previously been shown to be associated with dieldrin resistance in certain other laboratory strains originating from West Africa, there was no obvious association between inversion karyotype assortment and the resistance phenotype in the Obuasi population. In addition, polymerase chain reaction analysis indicated the presence of the alanine296 to glycine mutation in the GABA (γ amino-butyric acid) receptor (which has been mapped to a chromosomal position within inversion 2La). This mutation has previously been shown to be associated with dieldrin resistance in the same An. gambiae laboratory strains of West African origin. Our data show only a weak association between the dieldrin resistance phenotype and the presence of this mutation, suggesting that another dieldrin resistance mechanism is operational in the Obuasi population. Biochemical and synergist exposure assays suggest a metabolic component, probably mediated by monooxygenase P450 enzymes. We conclude that dieldrin resistance in the An. gambiae population of the Obuasi region occurs at a high level - most likely in the absence of selection - and that control of the resistance phenotype is polyfactorial and must include components other than mutations in the GABA receptor locus. [ABSTRACT FROM AUTHOR]

Point mutations in the voltage-gated sodium channel gene involved in knockdown resistance to DDT and pyrethroid insecticides have been described in several insect species. In the malaria vector Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) two mutations have been identified. The first, consisting of a leucine–phenylalanine substitution at amino acid position 1014, is widespread in West Africa. The second, a leucine–serine substitution at the same position, has to date only been detected in western Kenya. Analysis of the kdr polymorphism in a sample of 106 An. gambiae s.s. of the rDNA S-form/Type I collected in Libreville (Gabon) surprisingly revealed the presence of both East and West African kdr mutations with frequencies of 63% and 37%, respectively. No wild-type alleles were detected and there was an excess of heterozygous genotypes ( P = 0.04). In addition, an inconsistency was found during the kdr genotyping procedures by polymerase chain reaction, which could have lead to an underestimation of resistance alleles. The implications of these findings are discussed. [ABSTRACT FROM AUTHOR]

The influence of substrate moisture (hydration) and grain size (texture) on oviposition was quantified in choice tests using Anopheles gambiae sensu stricto Giles (Diptera: Culicidae) laboratory strains and gravid An. gambiae sensu lato from a natural population in Western Kenya. A strong, positive correlation was found between moisture content and the degree of egg-laying, which peaked at saturation with standing water. Soil moisture quantified as surface conductivity, was measured with an electronic leaf-wetness sensor slightly modified from a unit available commercially. Although An. gambiae females were sensitive to measurable differences in substrate moisture, they distributed eggs on both fully hydrated and less hydrated substrates. In contrast, An. gambiae females showed little response to substrate texture: they oviposited with equal frequency on all silica substrates of eight particle size classes, ranging from small pebbles (850 µm diameter) to very fine grains (< 38 µm diameter), when all were moist. Female An. gambiae laid more eggs on dark than white substrates against a light background, but did not discriminate between moist, pulverized black soapstone and moist black Kenyan soil taken from typical An. gambiae larval habitats. We conclude that hydration and visual contrast are critical ovipositional site qualities for An. gambiae, but substrate texture is not. [ABSTRACT FROM AUTHOR]

Mating by male Anopheles mosquitoes (Diptera: Culicidae) was evaluated in the laboratory to assess fitness effects of radio-sterilization applied during different life stages of the malaria vectors An. stephensi Liston and An. gambiae Giles sensu stricto. After reproductive sterilization by gamma-irradiation (120 Gy), equal proportions of sterile and fertile (unirradiated) male adults were released into cages with virgin females and allowed to compete for matings. Radio-sterilization was applied when the males were pupae aged 0–7 h or 24–32 h, or adults aged <24 h or 24–55 h. After being radio-sterilized in the adult stage, males of both species competed effectively with unirradiated males, whereas those sterilized in the pupal stage obtained significantly fewer matings than unirradiated males from the same cohort. There was no evidence of females obtaining multiple inseminations. These findings emphasize the need to radio-sterilize males as adults in order to minimize the fitness cost. Such males may be intended for sterile insect technique population suppression or for trial releases of transgenic anophelines. [ABSTRACT FROM AUTHOR]

The Ghanaian National Malaria Control Programme has prioritized insecticide-treated materials as a key strategy for malaria control. We report on a survey of the distribution of the molecular forms ofAnopheles gambiaeGiles (Diptera: Culicidae) and insecticide resistance (thekdrmutation), carried out by sampling mosquitoes from 11 locations in Ghana and one additional site in Burkina Faso. The molecular M and S forms ofAn. gambiaewere found to occur in sympatry in southern Ghana. The S form predominated throughout its distribution in the coastal savannah, except at one location in the strand and mangrove zone where rice was cultivated. The M form was the only form collected in northern Ghana and was the predominant form (97.5%) in Burkina Faso. No M/S hybrids were detected. Thekdrmutation was observed at very high frequencies (98–100%) within the S form but reached a maximum of only 3.38% in the M form in one population at an irrigation scheme in the Ghanaian coastal savannah zone. [ABSTRACT FROM AUTHOR]

Anopheline larval habitats associated with a swamp, were examined in a highland area (1910 m elevation) of western Kenya. A significant association was found between occurrence of Anopheles gambiae Giles s.s. (Diptera: Culicidae) larvae and two factors, habitat size and vegetation type. Over 80% of An. gambiae s.s. larvae were found in small isolated pools, characterized by short plants, occurring in both swamp margins and roadside ditches. However, Anopheles gambiae s.s. was not found in habitats marked by papyrus and floating plants. The larval habitat of An. gambiae s.s. was characterized by warmer daytime temperatures of water, which were significantly affected by habitat size and plant size. The density of indoor resting An. gambiae s.s. was 0.22 per house and negatively associated with distance from the swamp. These results indicate that the practice of swamp cultivation, in populated areas of the African highlands, increases availability and enhances habitat conditions for the malaria vector. [ABSTRACT FROM AUTHOR]

Vector abundance is an important factor governing disease risk and is often employed when modelling disease transmission. The longevity of the aquatic stages of mosquitoes (Diptera: Culicidae) dictates the rate of production of adults and hence the intensity of disease transmission. We examined how temperature influences the survival of larval stages (larvae and pupae) of Anopheles gambiae Giles sensu stricto and subsequent adult production of this most efficient malaria vector. Groups of 30 mosquitoes were reared at constant temperatures (from 10 to 40 °C) from the first instar and observed until death or metamorphosis of the last individual. Larvae developed into adults at temperatures ranging from 16 to 34 °C. Larval survival was shortest (< 7 days) at 10–12 °C and 38–40 °C, and longest (> 30 days) at 14–20 °C. Within the temperature range at which adults were produced, larval mortality was highest at the upper range 30–32 °C, with death (rather than adult emergence) representing over 70% of the terminal events. The optimal survival temperatures were lower than the temperatures at which development was quickest, suggesting a critical relationship between temperature and the life cycle of the insect. These data provide fundamental information about An. gambiae s.s. adult productivity at different temperatures, which may facilitate the construction of process-based models of malaria risk in Africa and the development of early warning systems for epidemics. [ABSTRACT FROM AUTHOR]