Anopheles Populations Research Articles

Exposure of malaria vector larval habitats to domestic pollutants escalate insecticides resistance: experimental proof

Poor domestic waste management in unplanned urban areas have resulted into formation of polluted potential malaria vector larval habitats. Most of these larval habitats are contaminated with domestic pollutants such as hydrogen peroxide and detergents. The present study was designed to evaluate the susceptibility status of Anopheles gambie and Anopheles arabiensis to commonly used insecticides after being exposed to hydrogen peroxide and soap detergents in northern Tanzania. Anopheles gambiae s.s larvae were collected from the laboratory colony whereas those for An. arabiensis were sampled from the rice field in lower Moshi. The larvae were reared in two arms, larval habitats in arm one had water containing soaps while in arm two, the larval habitats contained hydrogen peroxide at different concentrations (0.001%, 0.0001%,0.00001% and 0.000001%). Emerging adult’s female of the two Anopheles species from the two larval habitats were exposed to 0.75% and 0.05% of permethrin and deltamethrin WHO Insecticides impregnated standard papers. Knockdown rates were observed for 1 h and mortality rates within 24 h after exposure. Regression analysis was performed using probit analysis to establish knockdown rates and lethal concentration for 50% and 95%. The wild and laboratory anopheles populations reared in detergent or hydrogen peroxide in different concentrations showed reduced susceptibility to permethrin and deltamethrin. The median knockdown time for 50% of the population (KDT50) ranged between 13.52 min (10.98–19.74) to 30.67 min (28.53–34.34) while the KDT95 raised between 32.01 min (29.47–38.23) to 111.84 min (109.70–115.54) for those reared in detergent and exposed to deltamethrin and permethrin. For those reared in hydrogen peroxide and exposed to deltamethrin and permethrin, KDT50 ranged between 8.24 min (6.92–13.51) to 24.28 min(22.70–27.46) while KDT95 ranged between 25.94 min(24.62–31.21) to 118.68 min(117.10–121.86). Mortality of wild population exposed to standard insectides treated papers after reared in detergent ranged between 40 to 95.3% while for those reared in hydrogen peroxide ranged between 67 to 95.3%. The findings of this study demonstrate that domestic waste pollutants such as hydrogen peroxide and soaps are partly associated with reduced susceptibility to insecticides among potential malaria vectors.

A sensory appendage protein protects malaria vectors from pyrethroids.

SummaryPyrethroid-impregnated bednets have driven significant reductions in malaria morbidity and mortality in Africa since the beginning of the century 1. The intense selection pressure exerted by bednets has precipitated widespread and escalating pyrethroid resistance in African Anopheles populations, threatening to reverse gains made by malaria control 2. Here we show that a leg-enriched sensory appendage protein, SAP2, confers pyrethroid resistance in Anopheles gambiae. SAP2 expression is elevated in insecticide-resistant populations and is further induced upon mosquito contact with pyrethroids. SAP2 silencing fully restores mosquito mortality, whilst its overexpression results in increased resistance, likely due to the high-affinity SAP2 binding to pyrethroid insecticides. Mining of genome sequence data reveals a selective sweep near the SAP2 locus in three West African countries, with the observed increase in haplotype associated SNPs mirroring increasing resistance reported in Burkina Faso. Our study identifies a new insecticide resistance mechanism that is likely highly relevant to malaria control efforts.

Bio-efficacy and wash resistance of MAGNet long-lasting insecticidal net against wild populations of Anopheles funestus in experimental huts in Muheza, Tanzania

BackgroundThe decline in malaria cases and vectors is major milestone in fighting against malaria. The efficacy of MAGNet long-lasting insecticidal nets (MAGNet LLIN), an alpha-cypermethrin incorporated long-lasting net, with the target dose ± 25% of 5.8 g active ingredient (AI)/kg (4.35–7.25 g AI/kg) was evaluated in six veranda-trap experimental huts in Muheza, Tanzania against freely flying wild population of Anopheles funestus.MethodsMAGNet LLINs were tested against wild, free-flying, host-seeking An. funestus mosquitoes over a period of 6 weeks (total of 36 nights in the huts). MAGNet LLIN efficacy was determined in terms of mosquito mortality, blood-feeding inhibition, deterrence, induced exiting, personal protection, and insecticidal killing over 20 washes according to WHO standardized procedures. Efficacy was compared with reference to a WHOPES recommended approved LLINs (DuraNet) and to a net conventionally treated (CTN) treated with alpha-cypermethrin at WHO-recommended dose and washed to just before cut-off point. The efficacy of MAGNet was evaluated in experimental huts against wild, free-flying, pyrethroid-resistant An. funestus. The WHO-susceptibility method was used to detect resistance in wild Anopheles exposed to 0.75% permethrin. Mosquito mortality, blood-feeding inhibition and personal protection were compared between untreated nets and standard LLINs. Blood-feeding rates were recorded and compared between the 20 times washed; blood-feeding rates between 20 times washed MAGNet LLIN and 20 times washed WHOPES-approved piperonyl butoxide (PBO)/pyrethroid were not statistically different (p > 0.05).ResultsThe results have evidently shown that MAGNet LLIN provides similar blood-feeding inhibition, exophily, mortality, and deterrence to the standard approved LLIN, thus meeting the WHOPES criteria for blood feeding. The significantly high feeding inhibition and personal protection over pyrethroid-resistant An. funestus recorded by both unwashed and 20 times washed MAGNet compared to the unwashed DuraNet, the WHOPES-approved standard pyrethroid-only LLIN provides proof of MAGNet meeting Phase II WHOPES criteria for a LLIN.ConclusionBased on this study, MAGNet has been shown to have a promising impact on protection when 20 times washed against a highly resistant population of An. funestus.

Natural Wolbachia infections are common in the major malaria vectors in Central Africa.

During the last decade, the endosymbiont bacterium Wolbachia has emerged as a biological tool for vector disease control. However, for long time, it was believed that Wolbachia was absent in natural populations of Anopheles. The recent discovery that species within the Anopheles gambiae complex host Wolbachia in natural conditions has opened new opportunities for malaria control research in Africa. Here, we investigated the prevalence and diversity of Wolbachia infection in 25 African Anopheles species in Gabon (Central Africa). Our results revealed the presence of Wolbachia in 16 of these species, including the major malaria vectors in this area. The infection prevalence varied greatly among species, confirming that sample size is a key factor to detect the infection. Moreover, our sequencing and phylogenetic analyses showed the important diversity of Wolbachia strains that infect Anopheles. Co‐evolutionary analysis unveiled patterns of Wolbachia transmission within some Anopheles species, suggesting that past independent acquisition events were followed by co‐cladogenesis. The large diversity of Wolbachia strains that infect natural populations of Anopheles offers a promising opportunity to select suitable phenotypes for suppressing Plasmodium transmission and/or manipulating Anopheles reproduction, which in turn could be used to reduce the malaria burden in Africa.

Is Anopheles gambiae a Natural Host of Wolbachia?

Wolbachia (Alphaproteobacteria, Rickettsiales) is an intraovarially transmitted symbiont of insects able to exert striking phenotypes, including reproductive manipulations and pathogen blocking. These phenotypes make Wolbachia a promising tool to combat mosquito-borne diseases. Although Wolbachia is present in the majority of terrestrial arthropods, including many disease vectors, it was considered absent from Anopheles gambiae mosquitos, the main vectors of malaria in sub-Saharan Africa. In 2014, Wolbachia sequences were detected in A. gambiae samples collected in Burkina Faso. Subsequently, similar evidence came from collections all over Africa, revealing a high Wolbachia 16S rRNA sequence diversity, low abundance, and a lack of congruence between host and symbiont phylogenies. Here, we reanalyze and discuss recent evidence on the presence of Wolbachia sequences in A. gambiae. We find that although detected at increasing frequencies, the unusual properties of these Wolbachia sequences render them insufficient to diagnose natural infections in A. gambiae Future studies should focus on uncovering the origin of Wolbachia sequence variants in Anopheles and seeking sequence-independent evidence for this new symbiosis. Understanding the ecology of Anopheles mosquitos and their interactions with Wolbachia will be key in designing successful, integrative approaches to limit malaria spread. Although the prospect of using Wolbachia to fight malaria is intriguing, the newly discovered strains do not bring it closer to realization.IMPORTANCEAnopheles gambiae mosquitos are the main vectors of malaria, threatening around half of the world's population. The bacterial symbiont Wolbachia can interfere with disease transmission by other important insect vectors, but until recently, it was thought to be absent from natural A. gambiae populations. Here, we critically analyze the genomic, metagenomic, PCR, imaging, and phenotypic data presented in support of the presence of natural Wolbachia infections in A. gambiae We find that they are insufficient to diagnose Wolbachia infections and argue for the need of obtaining robust data confirming basic Wolbachia characteristics in this system. Determining the Wolbachia infection status of Anopheles is critical due to its potential to influence Anopheles population structure and Plasmodium transmission.

High Plasmodium infection and multiple insecticide resistance in a major malaria vector Anopheles coluzzii from Sahel of Niger Republic

BackgroundInformation on insecticide resistance and the mechanisms driving it in the major malaria vectors is grossly lacking in Niger Republic, thus hindering control efforts. To facilitate evidence-based malaria control, the role of Anopheles coluzzii population from southern Niger, in malaria transmission, its insecticides resistance profile and the molecular mechanisms driving the resistance were characterized.MethodsBlood fed female Anopheles gambiae sensu lato resting indoor were collected at Tessaoua, Niger. Source of blood was established using PCR and infection with Plasmodium determined using TaqMan assay. Resistance profile was established with the major public health insecticides, and resistance intensity determined with deltamethrin. Synergist assays were conducted with piperonyl butoxide and diethyl maleate. Presence of L1014F and L1014S knockdown resistance (kdr) mutations in the voltage-gated sodium channel (VGSC) was investigated using TaqMan genotyping, and strength of selection pressure acting on the Anopheles populations determined by assessing the genetic diversity of a fragment spanning exon-20 of the VGSC from alive and dead females.ResultsHigh human blood index (96%) and high Plasmodium falciparum infection (~ 13%) was observed in the An. coluzzii population. Also, a single mosquito was found infected with Plasmodium vivax. High pyrethroid and organochloride resistance was observed with mortalities of less than 20% for deltamethrin, permethrin, α-cypermethrin, and DDT. A high LD50 (156.65 min) was obtained for deltamethrin, with a resistance ratio of ~ 47.18 compared to the susceptible Ngoussou colony. Moderate carbamate resistance was observed, and a full susceptibility to organophosphates recorded. Synergist bioassays with piperonyl butoxide and diethyl maleate significantly recovered deltamethrin and DDT susceptibility, respectively implicating CYP450 s (mortality = 82%, χ2 = 84.51, p < 0.0001) and glutathione S-transferases (mortality = 58%, χ2 = 33.96, p < 0.001) in resistance. A high frequency of 1014F kdr mutation (82%) was established, with significant difference in genotype distribution associated with permethrin resistance [odds ratio = 7.71 (95% CI 2.43–14.53, χ2 = 13.67, p = 0.001]. Sequencing of intron-1 of the voltage-gated sodium channel (VGSC) revealed a low genetic diversity.ConclusionHigh pyrethroid resistance highlight the challenges to the effectiveness of the pyrethroids-based ITNs and indoor residual spraying (IRS) against An. coluzzii in Niger. The pyrethroids-synergists LLINs and organophosphate-based IRS maybe the alternatives for malaria control in southern Niger.

Surveillance of Anopheles populations in different periods in Suizhou City

To evaluate the effect of malaria vector control measures so as to provide the evidence for consolidating the control effectiveness of malaria and carrying out the vector surveillance in Suizhou City. The distributions of Anopheles sinensis and An. anthropophagus were investigated by the combination of retrospective review and field survey. The changes of density and population of mosquito vectors were investigated and compared among various years. From 1985 to 1996, both An. sinensis and An. anthropophagus were found in 18 towns, and An. anthropophagus mosquitoes accounted for 52.3% of the total Anopheles mosquitoes. Only An. sinensis mosquitoes were found in other 26 towns. In 2003 and 2004, An. anthropophagus mosquitoes were found in the original 18 towns, and they accounted for 47.0% and 38.1% respectively, but in 2005, An. anthropophagus mosquitoes were not found in this city. However, the density of An. sinensis presented an upward trend. An. anthropophagus population has gradually disappeared in Suizhou City, and currently, An. sinensis is the main Anopheles population, which might be the malaria vector in suitable conditions. Therefore, the surveillance and control of Anophe les vector should be strengthened in order to consolidate the achievements of malaria elimination.

Detection of alleles associated with resistance to chemical insecticide in the malaria vector Anopheles arabiensis in Santiago, Cabo Verde

BackgroundMosquitoes of the Anopheles gambiae complex are the main malaria vectors worldwide. Due to the lack of a vaccine to prevent malaria, the principal way to reduce the impact of this disease relies on the use of chemical insecticides to control its vectors. However, the intensive use of such compounds has led to the emergence of insecticide resistance in several Anopheles populations in Africa. This study aimed to investigate the presence of resistance alleles in an Anopheles arabiensis population from the City of Praia, capital of the Archipelago Cabo Verde, one of the countries on the World Health Organization list of countries that are on a path to eliminate local transmission of malaria.MethodsLarvae from the Anopheles genus were collected using a one-pint dipper in three areas of City of Praia. Larvae were fed and maintained until the emergence of adult mosquitoes, and these were morphologically identified. In addition, molecular identification was performed using IGS markers and all An. arabiensis samples were subjected to PCR to screen for mutations associated to resistance in the Ace-1, Nav and GSTE2 genes.ResultsFrom a total of 440 mosquitoes collected, 52.3% were morphologically identified as An. gambiae sensu lato (s.l.) and 46.7% as Anopheles pretoriensis. The molecular identification showed that 100% of the An. gambiae s.l. were An. arabiensis. The mutations G119S in the Ace-1 gene and L119F in the GSTE2 gene were screened but not found in any sample. However, sequencing analysis for GSTE2 revealed the presence of 37 haplotypes, 16 polymorphic sites and a high genetic diversity (π = 2.67). The L1014S mutation in the Nav (voltage-gated sodium channel gene) was detected at a frequency of 7.3%.ConclusionThis is the first study to investigate the circulation of insecticide resistance alleles in An. arabiensis from Cabo Verde. The circulation of the L1014S allele in the population of An. arabiensis in the city of Praia suggests that pyrethroid resistance may arise, be quickly selected, and may affect the process of malaria elimination in Cabo Verde. Molecular monitoring of resistance should continue in order to guide the development of strategies to be used in vector control in the study region.

Species abundance, composition, and nocturnal activity of female Anopheles (Diptera: Culicidae) in malaria-endemic villages of Papua New Guinea: assessment with barrier screen sampling

BackgroundCommunity composition of Anopheles mosquitoes, and their host-seeking and peridomestic behaviour, are important factors affecting malaria transmission. In this study, barrier screen sampling was used to investigate species composition, abundance, and nocturnal activity of Anopheles populations in villages of Papua New Guinea.MethodsMosquitoes were sampled from 6 pm to 6 am in five villages from 2012 to 2016. The barrier screens were positioned between the village houses and the perimeter of villages where cultivated and wild vegetation (“the bush”) grew thickly. Female Anopheles that rested on either village or bush side of the barrier screens, as they commuted into and out of the villages, were captured. Similarity in species composition among villages was assessed. Mosquitoes captured on village and bush sides of the barrier screens were sorted by feeding status and by hour of collection, and their numbers were compared using negative binomial generalized linear models.ResultsFemales of seven Anopheles species were present in the sample. Species richness ranged from four to six species per village, but relative abundance was highly uneven within and between villages, and community composition was similar for two pairs of villages and highly dissimilar in a fifth. For most Anopheles populations, more unfed than blood-fed mosquitoes were collected from the barrier screens. More blood-fed mosquitoes were found on the side of the barrier screens facing the village and relatively more unfed ones on the bush side, suggesting commuting behaviour of unfed host-seeking females into the villages from nearby bush and commuting of blood-fed females away from villages towards the bush. For most populations, the majority of host-seeking mosquitoes arrived in the village before midnight when people were active and unprotected from the mosquitoes by bed nets.ConclusionThe uneven distribution of Anopheles species among villages, with each site dominated by different species, even among nearby villages, emphasizes the importance of vector heterogeneity in local malaria transmission and control. Yet, for most species, nocturnal activity patterns of village entry and host seeking predominantly occurred before midnight indicating common behaviours across species and populations relative to human risk of exposure to Anopheles bites.

Small-scale field evaluation of the efficacy and residual effect of Fludora\xae Fusion (mixture of clothianidin and deltamethrin) against susceptible and resistant Anopheles gambiae populations from Benin, West Africa

BackgroundIn recognition of the threat of insecticide resistance in vectors of malaria, the WHO Global Malaria Programme recommends the development of an appropriate and comprehensive response to insecticide resistance. In principle, good resistance management practice requires the application of multiple insecticides of different modes of action, for example, in rotations and mixtures. Insecticides recommended by the World Health Organization for indoor residual spraying and long-lasting insecticide nets are limited. It is, therefore, judicious to prevent the rapid spread of insecticide resistance by evaluating new insecticides formulations with different modes of action and long residual effect.MethodsFludora® Fusion, a new neonicotinoid IRS formulation (a mixture of 500 g/kg clothianidin and 62.5 g/kg deltamethrin applied 200 mg ai/sqm + 25 mg ai/sqm, respectively) was tested. Small scale field evaluation of this product was conducted in the district of Dangbo in Benin, to compare its efficacy and residual effect on cement and mud walls against those of clothianidin 200 mg ai/sqm (WG 70) alone, and of deltamethrin 25 mg ai/sqm (WG 250) alone. WHO wall cone bioassays were conducted monthly with laboratory susceptible Anopheles “Kisumu” and wild Anopheles gambiae sensu stricto (s.s.) population from Dangbo. The induced mortality by each treatment per wall substrate for 24 h, 48 h, and 72 h post exposure were recorded every month and analysed.ResultsFludora® Fusion and clothianidin WG 70 showed mortality rates over 80% WHO bio-efficacy threshold on cement walls either with susceptible or resistant An. gambiae s.s. over a period of 10 and 9 months, respectively. Treatment with Fludora® Fusion and clothianidin WG 70 on the mud walls showed residual effect for 6 months and 5 months respectively against both susceptible and resistant mosquitoes. During the whole evaluation period, deltamethrin WG 250 showed mortality rates below 80% against resistant Anopheles population. Furthermore, the knock down rates observed with the Fludora® Fusion combination were significantly higher (p < 5%) than those induced by Clothiandin WG 70.ConclusionBoth the Fludora® Fusion combination and clothianidin alone showed very good and lasting efficacy for IRS against resistant Anopheles with some residual benefit provided by the combination. The residual efficacy of the Fludora® Fusion combination evaluated at 10 months shows this product is a good candidate for IRS interventions.

Transcriptomic meta-signatures identified in Anopheles gambiae populations reveal previously undetected insecticide resistance mechanisms

Increasing insecticide resistance in malaria-transmitting vectors represents a public health threat, but underlying mechanisms are poorly understood. Here, a data integration approach is used to analyse transcriptomic data from comparisons of insecticide resistant and susceptible Anopheles populations from disparate geographical regions across the African continent. An unbiased, integrated analysis of this data confirms previously described resistance candidates but also identifies multiple novel genes involving alternative resistance mechanisms, including sequestration, and transcription factors regulating multiple downstream effector genes, which are validated by gene silencing. The integrated datasets can be interrogated with a bespoke Shiny R script, deployed as an interactive web-based application, that maps the expression of resistance candidates and identifies co-regulated transcripts that may give clues to the function of novel resistance-associated genes.

A trade-off between dry season survival longevity and wet season high net reproduction can explain the persistence of Anopheles mosquitoes

BackgroundPlasmodium falciparum malaria remains a leading cause of death in tropical regions of the world. Despite efforts to reduce transmission, rebounds associated with the persistence of malaria vectors have remained a major impediment to local elimination. One area that remains poorly understood is how Anopheles populations survive long dry seasons to re-emerge following the onset of the rains.MethodsWe developed a suite of mathematical models to explore the impact of different dry-season mosquito survival strategies on the dynamics of vector populations. We fitted these models to an Anopheles population data set from Mali to estimate the model parameters and evaluate whether incorporating aestivation improved the fit of the model to the observed seasonal dynamics. We used the fitted models to explore the impact of intervention strategies that target aestivating mosquitoes in addition to targeting active mosquitoes and larvae.ResultsIncluding aestivation in the model significantly improved our ability to reproduce the observed seasonal dynamics of vector populations as judged by the deviance information criterion (DIC). Furthermore, such a model resulted in more biologically plausible active mosquito survival times (for A. coluzzii median wet season survival time of 10.9 days, 95% credible interval (CrI): 10.0–14.5 days in a model with aestivation versus 38.1 days, 95% CrI: 35.8–42.5 days in a model without aestivation; similar patterns were observed for A. arabiensis). Aestivation also generated enhanced persistence of the vector population over a wider range of both survival times and fecundity levels. Adding vector control interventions that target the aestivating mosquito population is shown to have the potential to enhance the impact of existing vector control.ConclusionsDry season survival attributes appear to drive vector population persistence and therefore have implications for vector control. Further research is therefore needed to better understand these mechanisms and to evaluate the additional benefit of vector control strategies that specifically target dormant mosquitoes.

Prevention of malaria transmission around reservoirs: an observational and modelling study on the effect of wind direction and village location

Many large dams are constructed annually in Africa, with associated reservoirs that might exacerbate the risk of malaria in new villages built nearby. We aimed to investigate the heterogeneous risk of malaria around reservoirs related to the impact of wind direction on malaria transmission. Between June 15, 2012, and April 22, 2015, we obtained field data on climate and hydrological conditions, and monitored Anopheles mosquito populations around the Koka reservoir in Ethiopia using in-situ weather stations, mosquito light traps, and larval dipping. The field data were used to calibrate a field-tested, spatially explicit mechanistic malaria transmission model, the Hydrology, Entomology, and Malaria Transmission Simulator (HYDREMATS), to investigate the effect of relative wind direction on malaria transmission and associated mechanisms. We combined our simulation results and observational data to assess the association between village location around a reservoir and risk of malaria. HYDREMATS simulations showed that wind blowing from a village towards a reservoir increases the size of malaria vector populations, whereas wind blowing from a reservoir towards a village decreases the size of malaria vector populations, which was consistent with our field data. Larval mortality is low in locations with village-to-reservoir wind due to weak surface waves, and this wind direction creates conditions that enable mosquitoes to identify village locations more easily than in conditions caused by reservoir-to-village wind, which increases the size of malaria vector populations, and thus malaria transmission. Among the wind conditions investigated (0·5-5 m/s), the effect of CO2 attraction on the size of the Anopheles population was largest at wind speeds of 0·5 m/s and 1 m/s, decreasing with higher wind speed. At a wind speed of 5 m/s, the effect of CO2 attraction was negligible, whereas the effect of waves was strongest. The effect of advection on Anopheles population size was negligible at all wind speeds and wind directions. The effect of wind on malaria transmission around reservoirs can be substantial. The transmission of malaria can be minimised if the location of villages situated near a reservoir is carefully considered. For areas in which the environmental conditions surrounding a resevoir are equal, villages should be located downwind of reservoirs to reduce the incidence of malaria, although further research will be required in locations where wind direction changes in different seasons. US National Science Foundation, and Cooperative Agreement between the Masdar Institute of Science and Technology, Abu Dhabi, UAE, and the Massachusetts Institute of Technology, Cambridge, MA, USA.

Opening the toolkit for genetic analysis and control of Anopheles mosquito vectors.

Anopheles is the only genus of mosquitoes that transmit human malaria and consequently the focus of large scale genome and transcriptome-wide association studies. Genetic tools to define the function of the candidate genes arising from these analyses are vital. Moreover, genome editing offers the potential to modify Anopheles population structure at local and global scale to provide complementary tools towards the ultimate goal of malaria elimination. Major breakthroughs in Anopheles genetic analysis came with the development of germline transformation and RNA interference technology. Yet, the field has been revolutionised again by precise genome editing now possible through site-specific nucleases. Here we review the components of the current genetic toolkit available to study Anopheles, focusing particularly on how these technical advances are used to gain insight into malaria transmission and the design of genetic methods to control Anopheles vectors.

Experimental huts trial of the efficacy of pyrethroids/piperonyl butoxide (PBO) net treatments for controlling multi-resistant populations of Anopheles funestus s.s. in Kpomè, Southern Benin

Background: Insecticides resistance in Anopheles mosquitoes limits Long-Lasting Insecticidal Nets (LLIN) used for malaria control in Africa, especially Benin. This study aimed to evaluate the bio-efficacy of current LLINs in an area where An. funestus s.l. and An. gambiae have developed multi-resistance to insecticides, and to assess in experimental huts the performance of a mixed combination of pyrethroids and piperonyl butoxide (PBO) treated nets on these resistant mosquitoes. Methods: The study was conducted at Kpomè, Southern Benin. The bio-efficacy of LLINs against An. funestus and An. gambiae was assessed using the World Health Organization (WHO) cone and tunnel tests. A released/recapture experiment following WHO procedures was conducted to compare the efficacy of conventional LLINs treated with pyrethroids only and LLINs with combinations of pyrethroids and PBO. Prior to huts trials, we confirmed the level of insecticide and PBO residues in tested nets using high performance liquid chromatography (HPLC). Results: Conventional LLINs (Type 2 and Type 4) have the lowest effect against local multi-resistant An. funestus s.s. and An. coluzzii populations from Kpomè. Conversely, when LLINs containing mixtures of pyrethroids and PBO (Type 1 and Type 3) were introduced in trial huts, we recorded a greater effect against the two mosquito populations (P < 0.0001). Tunnel test with An. funestus s.s. revealed mortalities of over 80% with this new generation of LLINs (Type 1 and Type 3),while conventional LLINs produced 65.53 ± 8.33% mortalities for Type 2 and 71.25 ±7.92% mortalities for Type 4. Similarly, mortalities ranging from 77 to 87% were recorded with the local populations of An. coluzzii. Conclusion: This study suggests the reduced efficacy of conventional LLINs (Pyrethroids alone) currently distributed in Benin communities where Anopheles populations have developed multi-insecticide resistance. The new generation nets (pyrethroids+PBO) proved to be more effective on multi-resistant populations of mosquitoes.

An assessment of mosquito collection techniques for xenomonitoring of anopheline-transmitted Lymphatic Filariasis in Ghana.

Monitoring vectors is relevant to ascertain transmission of lymphatic filariasis (LF). This may require the best sampling method that can capture high numbers of specific species to give indication of transmission. Gravid anophelines are good indicators for assessing transmission due to close contact with humans through blood meals. This study compared the efficiency of an Anopheles gravid trap (AGT) with other mosquito collection methods including the box and the Centres for Disease Control and Prevention gravid, light, exit and BioGent-sentinel traps, indoor resting collection (IRC) and pyrethrum spray catches across two endemic regions of Ghana. The AGT showed high trapping efficiency by collecting the highest mean number of anophelines per night in the Western (4.6) and Northern (7.3) regions compared with the outdoor collection methods. Additionally, IRC was similarly efficient in the Northern region (8.9) where vectors exhibit a high degree of endophily. AGT also showed good trapping potential for collecting Anopheles melas which is usually difficult to catch with existing methods. Screening of mosquitoes for infection showed a 0.80-3.01% Wuchereria bancrofti and 2.15-3.27% Plasmodium spp. in Anopheles gambiae. The AGT has shown to be appropriate for surveying Anopheles populations and can be useful for xenomonitoring for both LF and malaria.

Efficacy of a novel mode of action of an indoor residual spraying product, SumiShield\xae 50WG against susceptible and resistant populations of Anopheles gambiae (s.l.) in Benin, West Africa

BackgroundScale-up of the distribution of long-lasting insecticide-treated bed nets and indoor residual spraying with insecticides over the last decade have contributed to the considerable decrease of malaria morbidity and mortality in sub-Saharan Africa. Due to the increasing pyrethroid resistance intensity and the spread of carbamate resistance in Anopheles gambiae (s.s.) mosquitoes and the limited number of insecticides recommended by the WHO for vector control, alternative insecticide formulations for IRS with long-lasting residual activity are required to sustain the gains obtained in most malaria-endemic countries.MethodsSumiShield 50WG (clothianidin 300 mg ai/m2) developed by Sumitomo Chemical was evaluated alongside deltamethrin 25 mg ai/m2 (K-Othrine 250 WG) against a pyrethroid resistant Anopheles gambiae (s.l.) population in experimental huts in Covè, Benin. Residual activity was also tested in cone bioassays with the susceptible An. gambiae “Kisumu” strain and the local wild resistant population.ResultsThe results showed very low toxicity from deltamethrin (mortality rates ranged between 1–40%) against host-seeking resistant Anopheles populations. SumiShield in contrast gave an overall mean mortality of 91.7% at the 120 h observation across the eight- month observation period following spraying. The residual activity measured using cone tests was over the 80% WHO threshold for 24 weeks for resistant wild Anopheles population and 32 weeks for the susceptible strain “Kisumu” after the spraying.ConclusionsSumiShield is a good candidate for IRS in areas of permanent malaria transmission and where Anopheles populations are resistant to other conventional insecticides such as pyrethroids. It would be interesting to complete experimental huts studies by assessing the efficacy and residual effect of SumiShield 50WG at community level (small-scale field testing) in an area where vectors are highly resistant to insecticides.

Development of an autodissemination strategy for the deployment of novel control agents targeting the common malaria mosquito, Anopheles quadrimaculatus say (Diptera: Culicidae)

BackgroundThe reduced efficacy of current Anopheline mosquito control methods underscores the need to develop new methods of control that exploit unique target sites and/or utilizes novel deployment methods. Autodissemination methodologies using insect growth regulators (IGRs) is growing in interest and has been shown to be effective at controlling Aedes mosquitoes in semi-field and field environments, yet little information exists for Anopheline mosquitoes. Therefore, we tested the hypothesis that female-driven autodissemination of an IGR combined with a new mechanism of action insecticide (Kir channel inhibitor) could be employed to reduce Anopheline populations.MethodologyWe studied the ability of three IGRs to be transferred to the larval habitat during oviposition in laboratory and semi-field environments. Adult mosquitoes were exposed to the chemicals for 4 hours immediately after blood feeding and efficacy was tested using classical methodologies, including adult emergence inhibition and High Performance Liquid Chromatography (HPLC). A complete autodissemination design was tested in a semi-field environment.Principal findingsLarval survivability and adult emergence were significantly reduced in habitats that were visited by novaluron treated adults, but no statistical differences were observed with pyriproxyfen or triflumuron. These data suggested novaluron, but not pyriproxyfen or triflumuron, was horizontally transferred from the adult mosquito to the larval habitat during oviposition. HPLC studies supported the toxicity data and showed that novaluron was present in the majority of larval habitats, suggesting that novaluron can be horizontally transferred by Anopheles quadrimaculatus. Importantly, the combination of novaluron and the Kir channel inhibitor, VU041, was capable of reducing adult and larval populations in semi-field environments.ConclusionsNovaluron can be transferred to the adult at a greater efficacy and/or is not degraded as quickly during the gonotropic cycle when compared to pyriproxyfen or triflumuron. Pending field confirmation, autodissemination approaches with novaluron may be a suitable tool to manage Anopheles populations.

Insecticide resistance in Anopheles arabiensis populations from Dakar and its suburbs: role of target site and metabolic resistance mechanisms

BackgroundUrban malaria is an increasing concern in most of the sub-Saharan Africa countries. In Dakar, the capital city of Senegal, the malaria epidemiology has been complicated by recurrent flooding since 2005. The main vector control measure for malaria prevention in Dakar is the community use of long-lasting insecticide-treated nets. However, the increase of insecticide resistance reported in this area needs to be better understood for suitable resistance management. This study reports the situation of insecticide resistance and underlying mechanisms in Anopheles arabiensis populations from Dakar and its suburbs.ResultsAll the populations tested showed resistance to almost all insecticides except organophosphates families, which remain the only lethal molecules. Piperonil butoxide (PBO) and ethacrinic acid (EA) the two synergists used, have respectively and significantly restored the susceptibility to DDT and permethrin of Anopheles population. Molecular identification of specimens revealed the presence of An. arabiensis only. Kdr genotyping showed the presence of the L1014F mutation (kdr-West) as well as L1014S (kdr-East). This L1014S mutation was found at very high frequencies (89.53%) in almost all districts surveyed, and in association with the L1014F (10.24%).ConclusionResults showed the contribution of both target-site and metabolic mechanisms in conferring pyrethroid resistance to An. arabiensis from the flooded areas of Dakar suburbs. These data, although preliminary, stress the need for close monitoring of the urban An. arabiensis populations for a suitable insecticide resistance management system to preserve core insecticide-based vector control tools in this flooded area.

Environmental Determinants of Malaria Transmission Around the Koka Reservoir in Ethiopia.

New dam construction is known to exacerbate malaria transmission in Africa as the vectors of malaria—Anopheles mosquitoes—use bodies of water as breeding sites. Precise environmental mechanisms of how reservoirs exacerbate malaria transmission are yet to be identified. Understanding of these mechanisms should lead to a better assessment of the impacts of dam construction and to new prevention strategies. Combining extensive multiyear field surveys around the Koka Reservoir in Ethiopia and rigorous model development and simulation studies, environmental mechanisms of malaria transmission around the reservoir were examined. Most comprehensive and detailed malaria transmission model, Hydrology, Entomology, and Malaria Transmission Simulator, was applied to a village adjacent to the reservoir. Significant contributions to the dynamics of malaria transmission are shaped by wind profile, marginal pools, temperature, and shoreline locations. Wind speed and wind direction influence Anopheles populations and malaria transmission during the major and secondary mosquito seasons. During the secondary mosquito season, a noticeable influence was also attributed to marginal pools. Temperature was found to play an important role, not so much in Anopheles population dynamics, but in malaria transmission dynamics. Change in shoreline locations drives malaria transmission dynamics, with closer shoreline locations to the village making malaria transmission more likely. Identified environmental mechanisms help in predicting malaria transmission seasons and in developing village relocation strategies upon dam construction to minimize the risk of malaria.