Malaria Vector Mosquito Research Articles

Larvicidal potential of carvacrol and terpinen-4-ol from the essential oil of Origanum vulgare (Lamiaceae) against Anopheles stephensi, Anopheles subpictus, Culex quinquefasciatus and Culex tritaeniorhynchus (Diptera: Culicidae)

Mosquito-borne diseases represent a deadly threat for millions of people worldwide. However, the use of synthetic insecticides to control Culicidae may lead to resistance, high operational costs and adverse non-target effects. Nowadays, plant-borne mosquitocides may serve as suitable alternative in the fight against mosquito vectors. In this study, the mosquito larvicidal activity of Origanum vulgare (Lamiaceae) leaf essential oil (EO) and its major chemical constituents was evaluated against the malaria vectors Anopheles stephensi and An. subpictus, the filariasis vector Culex quinquefasciatus and the Japanese encephalitis vector Cx. tritaeniorhynchus. The chemical composition of the EO was analyzed by gas chromatography–mass spectroscopy. GC–MS revealed that the essential oil of O. vulgare contained 17 compounds. The major chemical components were carvacrol (38.30%) and terpinen-4-ol (28.70%). EO had a significant toxic effect against early third-stage larvae of An. stephensi, An. subpictus, Cx. quinquefasciatus and Cx. tritaeniorhynchus, with LC50 values of 67.00, 74.14, 80.35 and 84.93μg/ml. The two major constituents extracted from the O. vulgare EO were tested individually for acute toxicity against larvae of the four mosquito vectors. Carvacrol and terpinen-4-ol appeared to be most effective against An. stephensi (LC50=21.15 and 43.27μg/ml, respectively) followed by An. subpictus (LC50=24.06 and 47.73μg/ml), Cx. quinquefasciatus (LC50=26.08 and 52.19μg/ml) and Cx. tritaeniorhynchus (LC50=27.95 and 54.87μg/ml). Overall, this research adds knowledge to develop newer and safer natural larvicides against malaria, filariasis and Japanese encephalitis mosquito vectors.

Highly efficient Cas9-mediated gene drive for population modification of the malaria vector mosquito Anopheles stephensi

Genetic engineering technologies can be used both to create transgenic mosquitoes carrying antipathogen effector genes targeting human malaria parasites and to generate gene-drive systems capable of introgressing the genes throughout wild vector populations. We developed a highly effective autonomous Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-associated protein 9 (Cas9)-mediated gene-drive system in the Asian malaria vector Anopheles stephensi, adapted from the mutagenic chain reaction (MCR). This specific system results in progeny of males and females derived from transgenic males exhibiting a high frequency of germ-line gene conversion consistent with homology-directed repair (HDR). This system copies an ∼ 17-kb construct from its site of insertion to its homologous chromosome in a faithful, site-specific manner. Dual anti-Plasmodium falciparum effector genes, a marker gene, and the autonomous gene-drive components are introgressed into ∼ 99.5% of the progeny following outcrosses of transgenic lines to wild-type mosquitoes. The effector genes remain transcriptionally inducible upon blood feeding. In contrast to the efficient conversion in individuals expressing Cas9 only in the germ line, males and females derived from transgenic females, which are expected to have drive component molecules in the egg, produce progeny with a high frequency of mutations in the targeted genome sequence, resulting in near-Mendelian inheritance ratios of the transgene. Such mutant alleles result presumably from nonhomologous end-joining (NHEJ) events before the segregation of somatic and germ-line lineages early in development. These data support the design of this system to be active strictly within the germ line. Strains based on this technology could sustain control and elimination as part of the malaria eradication agenda.

Insecticide resistance in the malaria vector Anopheles arabiensis in Mamfene, KwaZulu-Natal

The control of malaria vector mosquitoes in South Africa’s affected provinces is primarily based on indoor spraying of long-lasting residual insecticides. The primary vectors in South Africa are Anopheles arabiensis and An. funestus. South Africa’s National Malaria Control Programme has adopted a malaria elimination agenda and has scaled up vector control activities accordingly. However, despite these plans, local transmission continues and is most likely because of outdoor feeding by populations of An. arabiensis. An outdoor Anopheles surveillance system has been set up in three sections of the Mamfene district in northern KwaZulu- Natal in order to assess the extent of outdoor resting An. arabiensis in Mamfene and to assess the current insecticide susceptibility status of this population. According to WHO criteria, the An. arabiensis samples tested showed evidence of resistance to deltamethrin (pyrethroid), DDT (organochlorine) and bendiocarb (carbamate), and full susceptibility to the organophosphates pirimiphos-methyl and fenitrothion. Pre-exposure to piperonyl butoxide completely nullified the deltamethrin resistance otherwise evident in these samples, supporting previous studies implicating monooxygenase-based detoxification as the primary mechanism of pyrethroid resistance. The data presented here affirm the presence of pyrethroid and DDT resistance previously detected in this population and also indicate the comparatively recent emergence of resistance to the carbamate insecticide bendiocarb. These data show that special attention and commitment needs to be given to the principles of insecticide resistance management as well as to investigations into alternative control techniques designed to target outdoor-resting An. arabiensis in northern KwaZulu-Natal.

The fucomic potential of mosquitoes: Fucosylated N-glycan epitopes and their cognate fucosyltransferases

Fucoconjugates are key mediators of protein-glycan interactions in prokaryotes and eukaryotes. As examples, N-glycans modified with the non-mammalian core α1,3-linked fucose have been detected in various organisms ranging from plants to insects and are immunogenic in mammals. The rabbit polyclonal antibody raised against plant horseradish peroxidase (anti-HRP) is able to recognize the α1,3-linked fucose epitope and is also known to specifically stain neural tissues in the fruit fly Drosophila melanogaster. In this study, we have detected and localized the anti-HRP cross-reactivity in another insect species, the malaria mosquito vector Anopheles gambiae. We were able to identify and structurally elucidate fucosylated N-glycans including core mono- and difucosylated structures (responsible for anti-HRP cross reactivity) as well as a Lewis-type antennal modification on mosquito anionic N-glycans by applying enzymatic and chemical treatments. The three mosquito fucosyltransferase open reading frames (FucT6, FucTA and FucTC) required for the in vivo biosynthesis of the fucosylated N-glycan epitopes were identified in the Anopheles gambiae genome, cloned and recombinantly expressed in Pichia pastoris. Using a robust MALDI-TOF MS approach, we characterised the activity of the three recombinant fucosyltransferases in vitro and demonstrate that they share similar enzymatic properties as compared to their homologues from D. melanogaster and Apis mellifera. Thus, not only do we confirm the neural reactivity of anti-HRP in a mosquito species, but also demonstrate enzymatic activity for all its α1,3- and α1,6-fucosyltransferase homologues, whose specificity matches the results of glycomic analyses.

Eugenol, α-pinene and β-caryophyllene from Plectranthus barbatus essential oil as eco-friendly larvicides against malaria, dengue and Japanese encephalitis mosquito vectors.

Mosquito-borne diseases represent a deadly threat for millions of people worldwide. Eco-friendly mosquitocides are a priority. In Ayurvedic medicine, Plectranthus species have been used to treat heart disease, convulsions, spasmodic pain and painful urination. In this research, we evaluated the acute toxicity of essential oil from Plectranthus barbatus and its major constituents, against larvae of the malaria vector Anopheles subpictus, the dengue vector Aedes albopictus and the Japanese encephalitis vector Culex tritaeniorhynchus. The chemical composition of P. barbatus essential oil was analyzed by gas chromatography-mass spectroscopy. Nineteen components were identified. Major constituents were eugenol (31.12%), α-pinene (19.38%) and β-caryophyllene (18.42%). Acute toxicity against early third-instar larvae of An. subpictus, Ae. albopictus and Cx. tritaeniorhynchus was investigated. The essential oil had a significant toxic effect against larvae of An. subpictus, Ae. albopictus and Cx. tritaeniorhynchus, with 50% lethal concentration (LC50) values of 84.20, 87.25 and 94.34 μg/ml and 90% lethal concentration (LC90) values of 165.25, 170.56 and 179.58 μg/ml, respectively. Concerning major constituents, eugenol, α-pinene and β-caryophyllene appeared to be most effective against An. subpictus (LC50 = 25.45, 32.09 and 41.66 μg/ml, respectively), followed by Ae. albopictus (LC50 = 28.14, 34.09 and 44.77 μg/ml, respectively) and Cx. tritaeniorhynchus (LC50 = 30.80, 36.75 and 48.17 μg/ml, respectively). Overall, the chance to use metabolites from P. barbatus essential oil against mosquito vectors seems promising, since they are effective at low doses and could be an advantageous alternative to build newer and safer mosquito control tools.

A new class of insecticide for malaria vector control: evaluation of mosquito nets treated singly with indoxacarb (oxadiazine) or with a pyrethroid mixture against Anopheles gambiae and Culex quinquefasciatus.

BackgroundUniversal coverage with long-lasting insecticidal mosquito nets (LLIN) or indoor residual spraying (IRS) of houses remain the primary strategies for the control of mosquito vectors of malaria. Pyrethroid resistant malaria vectors are widespread throughout sub-Saharan Africa and new insecticides with different modes of action are urgently needed if malaria vector control is to remain effective. Indoxacarb is an oxadiazine insecticide that is effective as an oral and contact insecticide against a broad spectrum of agricultural pests and, due to its unique site of action, no cross-resistance has been detected through mechanisms associated with resistance to insecticides currently used in public health.MethodsWHO tunnel tests of host seeking mosquitoes were carried out as a forerunner to experimental hut trials, to provide information on dosage-dependent mortality, repellency, and blood-feeding inhibition. A dosage range of indoxacarb treated netting (100–1000 mg/m2) was tested against a pyrethroid susceptible strain of Anopheles gambiae. In addition, efficacy of indoxacarb 500 mg/m2 was compared with a standard pyrethroid formulation against pyrethroid susceptible and resistant Culex quinquefasciatus. Dosages between 25 and 300 mg/m2 indoxacarb were tested in tunnel tests and in ball-frame bioassays as mixtures with alphacypermethrin 25 mg/m2 and were compared with singly applied treatments against an insectary reared pyrethroid resistant strain of Cx. quinquefasciatus originally collected in Cotonou, Benin.ResultsThere was a dosage-dependent response in terms of indoxacarb induced mortality, with dosages >100 mg/m2 producing the best mortality response. In tunnel tests indoxacarb 500 mg/m2 exceeded WHOPES thresholds with >80 % mortality of adult An. gambiae and blood-feeding inhibition of 75 %. No cross-resistance to indoxacarb was detected through mechanisms associated with resistance to pyrethroid insecticides and was equally effective against susceptible and resistant strains of Cx. quinquefasciatus. Indoxacarb 500 mg/m2 killed 75 % of pyrethroid resistant Cx. quinquefasciatus compared with only 21 % mortality with alphacypermethrin 40 mg/m2. Mixtures of indoxacarb with pyrethroid produced an additive response for both mortality and blood-feeding inhibition. The best performing mixture (indoxacarb 200 mg/m2 + alphacypermethrin 25 mg/m2) killed 83 % of pyrethroid resistant Cx. quinquefasciatus and reduced blood-feeding by 88 %, while alphacypermethrin only killed 36 % and inhibited blood-feeding by 50 %.ConclusionsNew insecticides with different modes of action to those currently used in mosquito vector control are urgently needed. Indoxacarb shows great promise as a mixture with a pyrethroid and should be evaluated in experimental hut trials to determine performance against wild free-flying, pyrethroid resistant An. gambiae and wash-resistant formulations developed.

Seasonality of Plasmodium falciparum transmission: a systematic review.

BackgroundAlthough Plasmodium falciparum transmission frequently exhibits seasonal patterns, the drivers of malaria seasonality are often unclear. Given the massive variation in the landscape upon which transmission acts, intra-annual fluctuations are likely influenced by different factors in different settings. Further, the presence of potentially substantial inter-annual variation can mask seasonal patterns; it may be that a location has “strongly seasonal” transmission and yet no single season ever matches the mean, or synoptic, curve. Accurate accounting of seasonality can inform efficient malaria control and treatment strategies. In spite of the demonstrable importance of accurately capturing the seasonality of malaria, data required to describe these patterns is not universally accessible and as such localized and regional efforts at quantifying malaria seasonality are disjointed and not easily generalized.Methods The purpose of this review was to audit the literature on seasonality of P. falciparum and quantitatively summarize the collective findings. Six search terms were selected to systematically compile a list of papers relevant to the seasonality of P. falciparum transmission, and a questionnaire was developed to catalogue the manuscripts.Results and discussion152 manuscripts were identified as relating to the seasonality of malaria transmission, deaths due to malaria or the population dynamics of mosquito vectors of malaria. Among these, there were 126 statistical analyses and 31 mechanistic analyses (some manuscripts did both).DiscussionIdentified relationships between temporal patterns in malaria and climatological drivers of malaria varied greatly across the globe, with different drivers appearing important in different locations. Although commonly studied drivers of malaria such as temperature and rainfall were often found to significantly influence transmission, the lags between a weather event and a resulting change in malaria transmission also varied greatly by location.ConclusionsThe contradicting results of studies using similar data and modelling approaches from similar locations as well as the confounding nature of climatological covariates underlines the importance of a multi-faceted modelling approach that attempts to capture seasonal patterns at both small and large spatial scales.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0849-2) contains supplementary material, which is available to authorized users.

Pilot study on the combination of an organophosphate-based insecticide paint and pyrethroid-treated long lasting nets against pyrethroid resistant malaria vectors in Burkina Faso

A pilot study to test the efficacy of combining an organophosphate-based insecticide paint and pyrethroid-treated Long Lasting Insecticide Treated Nets (LLINs) against pyrethroid-resistant malaria vector mosquitoes was performed in a real village setting in Burkina Faso. Paint Inesfly 5A IGR™, comprised of two organophosphates (OPs) and an Insect Growth Regulator (IGR), was tested in combination with pyrethroid-treated LLINs. Efficacy was assessed in terms of mortality for 12 months using Early Morning Collections of malaria vectors and 30-minute WHO bioassays. Resistance to pyrethroids and OPs was assessed by detecting the frequency of L1014F and L1014S kdr mutations and Ace-1RG119S mutation, respectively. Blood meal origin was identified using a direct enzyme-linked immunosorbent assay (ELISA). The combination of Inesfly 5A IGR™ and LLINs was effective in killing 99.9–100% of malaria vector populations for 6 months regardless of the dose and volume treated. After 12 months, mortality rates decreased to 69.5–82.2%. The highest mortality rates observed in houses treated with 2 layers of insecticide paint and a larger volume. WHO bioassays supported these results: mortalities were 98.8–100% for 6 months and decreased after 12 months to 81.7–97.0%. Mortality rates in control houses with LLINs were low. Collected malaria vectors consisted exclusively of Anopheles coluzzii and were resistant to pyrethroids, with a L1014 kdr mutation frequency ranging from 60 to 98% through the study. About 58% of An. coluzzii collected inside houses had bloodfed on non-human animals. Combining Inesfly 5A IGR™ and LLINs yielded a one year killing efficacy against An. coluzzii highly resistant to pyrethroids but susceptible to OPs that exhibited an anthropo-zoophilic behaviour in the study area. The results obtained in a real setting supported previous work performed in experimental huts and underscore the need to study the impact that this novel strategy may have on clinical malaria and malaria exposure in children in a similar area of high pyrethroid resistance in South-Western Burkina Faso.

Genome-wide QTL mapping of saltwater tolerance in sibling species of Anopheles (malaria vector) mosquitoes

Although freshwater (FW) is the ancestral habitat for larval mosquitoes, multiple species independently evolved the ability to survive in saltwater (SW). Here, we use quantitative trait locus (QTL) mapping to investigate the genetic architecture of osmoregulation in Anopheles mosquitoes, vectors of human malaria. We analyzed 1134 backcross progeny from a cross between the obligate FW species An. coluzzii, and its closely related euryhaline sibling species An. merus. Tests of 2387 markers with Bayesian interval mapping and machine learning (random forests) yielded six genomic regions associated with SW tolerance. Overlap in QTL regions from both approaches enhances confidence in QTL identification. Evidence exists for synergistic as well as disruptive epistasis among loci. Intriguingly, one QTL region containing ion transporters spans the 2Rop chromosomal inversion that distinguishes these species. Rather than a simple trait controlled by one or a few loci, our data are most consistent with a complex, polygenic mode of inheritance.

Outdoor biting by Anopheles mosquitoes on Bioko Island does not currently impact on malaria control.

BackgroundThere have been many recent reports that the rate of outdoor biting by malaria vectors has increased. This study examined the impact this might have on malaria transmission by assessing the association between exposure to outdoor bites and malaria infection on Bioko Island, Equatorial Guinea.MethodsResponses to questions about time spent outside the previous night from a malaria indicator survey were combined with human landing catch measurements of hourly rates of outdoor and indoor biting for the whole island to estimate the number of outdoor and indoor bites received by each survey respondent. The association between RDT measured malaria infection status of individuals and outdoor bites received was investigated.ResultsThe average number of bites received per person per night was estimated as 3.51 in total, of which 0.69 (19.7%) would occur outdoors. Malaria infection was not significantly higher in individuals who reported spending time outside between 7 pm and 6 am the previous night compared to those not spending time outside in both adults (18.9% vs 17.4%, p = 0.20) and children (29.2% vs 27.1%, p = 0.20). Malaria infection in neither adults (p = 0.56) nor in children (p = 0.12) was associated with exposure to outdoor bites, even after adjusting for confounders.ConclusionsMalaria vector mosquitoes in Bioko do bite humans outdoors, and this has the potential to reduce the effectiveness of vector control. However, outdoor biting is currently not a major factor influencing the malaria burden, mainly because more than 95% of the population are indoors during the middle of the night, which is the peak biting period for malaria vector mosquitoes. The majority of resources should remain with control measures that target indoor biting and resting such as LLINs and IRS.

Endogenous siRNAs and piRNAs derived from transposable elements and genes in the malaria vector mosquito Anopheles gambiae.

BackgroundThe siRNA and piRNA pathways have been shown in insects to be essential for regulation of gene expression and defence against exogenous and endogenous genetic elements (viruses and transposable elements). The vast majority of endogenous small RNAs produced by the siRNA and piRNA pathways originate from repetitive or transposable elements (TE). In D. melanogaster, TE-derived endogenous siRNAs and piRNAs are involved in genome surveillance and maintenance of genome integrity. In the medically relevant malaria mosquito Anopheles gambiae TEs constitute 12-16% of the genome size. Genetic variations induced by TE activities are known to shape the genome landscape and to alter the fitness in An. gambiae.ResultsHere, using bioinformatics approaches we analyzed the small RNA data sets from 6 libraries formally reported in a previous study and examined the expression of the mixed germline/somatic siRNAs and piRNAs produced in adult An. gambiae females. We characterized a large population of TE-derived endogenous siRNAs and piRNAs, which constitutes 56-60% of the total siRNA and piRNA reads in the analysed libraries. Moreover, we identified a number of protein coding genes producing gene-specific siRNAs and piRNAs that were generally expressed at much lower levels than the TE-associated small RNAs. Detailed sequence analysis revealed that An. gambiae piRNAs were produced by both “ping-pong” dependent (TE-associated piRNAs) and independent mechanisms (genic piRNAs). Similarly to D. melanogaster, more than 90% of the detected piRNAs were produced from TE-associated clusters in An. gambiae. We also found that biotic stress as blood feeding and infection with Plasmodium parasite, the etiological agent of malaria, modulated the expression levels of the endogenous siRNAs and piRNAs in An. gambiae.ConclusionsWe identified a large and diverse set of the endogenously derived siRNAs and piRNAs that share common and distinct aspects of small RNA expression across insect species, and inferred their impact on TE and gene activity in An. gambiae. The TE-specific small RNAs produced by both the siRNA and piRNA pathways represent an important aspect of genome stability and genetic variation, which might have a strong impact on the evolution of the genome and vector competence in the malaria mosquitoes.Electronic supplementary materialThe online version of this article (doi:10.1186/s12864-015-1436-1) contains supplementary material, which is available to authorized users.

Understanding the long-lasting attraction of malaria mosquitoes to odor baits.

The use of odor baits for surveillance and control of malaria mosquitoes requires robust dispensing tools. In this study, the residual activity of a synthetic mosquito attractant blend dispensed from nylon or low density polyethylene (LDPE) sachets was evaluated at weekly intervals for one year without re-impregnation. The potential role of bacteria in modulating the attraction of mosquitoes to odor-treated nylon that had been used repeatedly over the one year study period, without re-impregnation, was also investigated. Significantly higher proportions of female Anopheles gambiae sensu stricto mosquitoes were consistently attracted to treated nylon strips than the other treatments, up to one year post-treatment. Additional volatile organic compounds and various bacterial populations were found on the treated nylon strips after one year of repeated use. The most abundant bacteria were Bacillus thuringiensis and Acinetobacter baumannii. Autoclaving of treated nylon strips prior to exposure had no effect on trap collections of laboratory-reared female An. Gambiae (P = 0.17) or wild female An. Gambiae sensu lato (P = 0.26) and Mansonia spp. (P = 0.17) mosquitoes. Trap catches of wild female An. Funestus (P < 0.001) and other anophelines (P < 0.007) were higher when treated strips had been autoclaved prior to deployment as opposed to when the treated nylon strips were not autoclaved. By contrast, wild female Culex mosquitoes were more strongly attracted to non-autoclaved compared to autoclaved treated nylon strips (P < 0.042). This study demonstrates the feasibility of using odor baits for sampling and surveillance of malaria as well as other mosquito vectors over prolonged periods of time. Preliminary evidence points towards the potential role of bacteria in sustaining prolonged use of nylon material for dispensing synthetic attractant odorants for host-seeking malaria and other mosquito vectors but further investigations are required.

Triosephosphate isomerase gene promoter variation: -5G/A and -8G/A polymorphisms in clinical malaria groups in two African populations

TPI1 promoter polymorphisms occur in high prevalence in individuals from African origin. Malaria-patients from Angola and Mozambique were screened for the TPI1 gene promoter variants rs1800200A>G, (-5G>A), rs1800201G>A, (-8G>A), rs1800202T>G, (-24T>G), and for the intron 5 polymorphism rs2071069G>A, (2262G>A). -5G>A and -8G>A variants occur in 47% and 53% in Angola and Mozambique, respectively while -24T>G was monomorphic for the wild-type T allele. Six haplotypes were identified and -8A occurred in 45% of the individuals, especially associated with the GAG haplotype and more frequent in non-severe malaria groups, although not significantly. The arising and dispersion of -5G>A and -8G>A polymorphisms is controversial. Their age was estimated by analyses of two microsatellite loci, CD4 and ATN1, adjacent to TPI1 gene. The -5G>A is older than -8G>A, with an average estimate of approximately 35,000years. The -8A variant arose in two different backgrounds, suggesting independent mutational events. The first, on the -5G background, may have occurred in East Africa around 20,800years ago; the second, on the -5A background, may have occurred in West Africa some 7500years ago. These estimates are within the period of spread of agriculture and the malaria mosquito vector in Africa, which could has been a possible reason for the selection of -8A polymorphism in malaria endemic countries.

The distinctive features of Indian malaria parasites

Malaria and factors driving malaria are heterogeneous in India, unlike in other countries, and the epidemiology of malaria therefore is considered 'highly complex'. This complexity is primarily attributed to several unique features of the malaria parasites, mosquito vectors, malaria-susceptible populations, and ecoclimatic variables in India. Recent research on the genetic epidemiology of Indian malaria parasites has been successful in partly unraveling the mysteries underlying these complexities.

Synthesis and characterization of silver nanoparticles using Gmelina asiatica leaf extract against filariasis, dengue, and malaria vector mosquitoes.

Mosquitoes are blood-feeding insects and serve as the most important vectors for spreading human diseases such as malaria, yellow fever, dengue fever, and filariasis. The continued use of synthetic insecticides has resulted in resistance in mosquitoes. Synthetic insecticides are toxic and affect the environment by contaminating soil, water, and air, and then natural products may be an alternative to synthetic insecticides because they are effective, biodegradable, eco-friendly, and safe to environment. Botanical origin may serve as suitable alternative biocontrol techniques in the future. The present study was carried out to establish the larvicidal potential of leaf extracts of Gmelina asiatica and synthesized silver nanoparticles using aqueous leaf extract against late third instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. Larvae were exposed to varying concentrations of plant extracts and synthesized AgNPs for 24 h. The results were recorded from UV-visible spectroscopy, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis support the biosynthesis and characterization of AgNPs. The maximum efficacy was observed in synthesized AgNPs against the larvae of An. stephensi (lethal dose (LC₅₀) = 22.44 μg/mL; LC₉₀ 40.65 μg/mL), Ae. aegypti (LC₅₀ = 25.77 μg/mL; LC₉₀ 45.98 μg/mL), and C. quinquefasciatus (LC₅₀ = 27.83 μg/mL; LC₉₀ 48.92 μg/mL), respectively. No mortality was observed in the control. This is the first report on mosquito larvicidal activity of plant-synthesized nanoparticles. Thus, the use of G. asiatica to synthesize silver nanoparticles is a rapid, eco-friendly, and a single-step approach and the AgNps formed can be potential mosquito larvicidal agents.

Underestimation of foraging behaviour by standard field methods in malaria vector mosquitoes in southern Africa.

BackgroundDefining the anopheline mosquito vectors and their foraging behaviour in malaria endemic areas is crucial for disease control and surveillance. The standard protocol for molecular identification of host blood meals in mosquitoes is to morphologically identify fed mosquitoes and then perform polymerase chain reaction (PCR), precipitin tests, or ELISA assays. The purpose of this study was to determine the extent to which the feeding rate and human blood indices (HBIs) of malaria vectors were underestimated when molecular confirmation by PCR was performed on both visually fed and unfed mosquitoes.MethodsIn association with the Southern Africa International Centers of Excellence in Malaria Research (ICEMR), mosquito collections were performed at three sites: Choma district in southern Zambia, Nchelenge district in northern Zambia, and Mutasa district in eastern Zimbabwe. All anophelines were classified visually as fed or unfed, and tested for blood meal species using PCR methods. The HBIs of visually fed mosquitoes were compared to the HBIs of overall PCR confirmed fed mosquitoes by Pearson’s Chi-Square Test of Independence.ResultsThe mosquito collections consisted of Anopheles arabiensis from Choma, Anopheles funestus s.s., Anopheles gambiae s.s. and Anopheles leesoni from Nchelenge, and An. funestus s.s. and An. leesoni from Mutasa. The malaria vectors at all three sites had large human blood indices (HBI) suggesting high anthropophily. When only visually fed mosquitoes tested by PCR for blood meal species were compared to testing those classified as both visually fed and unfed mosquitoes, it was found that the proportion blooded was underestimated by up to 18.7%. For most Anopheles species at each site, there was a statistically significant relationship (P < 0.05) between the HBIs of visually fed mosquitoes and that of the overall PCR confirmed fed mosquitoes.ConclusionThe impact on HBI of analysing both visually fed and unfed mosquitoes varied from site to site. This discrepancy may be due to partial blood feeding behaviour by mosquitoes, digestion of blood meals, sample condition, and/or expertise of entomology field staff. It is important to perform molecular testing on all mosquitoes to accurately characterize vector feeding behaviour and develop interventions in malaria endemic areas.

Biting behaviour of African malaria vectors: 1. where do the main vector species bite on the human body?

BackgroundMalaria control in Africa relies heavily on indoor vector management, primarily indoor residual spraying and insecticide treated bed nets. Little is known about outdoor biting behaviour or even the dynamics of indoor biting and infection risk of sleeping household occupants. In this paper we explore the preferred biting sites on the human body and some of the ramifications regarding infection risk and exposure management.MethodsWe undertook whole-night human landing catches of Anopheles arabiensis in South Africa and Anopheles gambiae s.s. and Anopheles funestus in Uganda, for seated persons wearing short sleeve shirts, short pants, and bare legs, ankles and feet. Catches were kept separate for different body regions and capture sessions. All An. gambiae s.l. and An. funestus group individuals were identified to species level by PCR.ResultsThree of the main vectors of malaria in Africa (An. arabiensis, An. gambiae s.s. and An. funestus) all have a preference for feeding close to ground level, which is manifested as a strong propensity (77.3% – 100%) for biting on lower leg, ankles and feet of people seated either indoors or outdoors, but somewhat randomly along the lower edge of the body in contact with the surface when lying down. If the lower extremities of the legs (below mid-calf level) of seated people are protected and therefore exclude access to this body region, vector mosquitoes do not move higher up the body to feed at alternate body sites, instead resulting in a high (58.5% - 68.8%) reduction in biting intensity by these three species.ConclusionsProtecting the lower limbs of people outdoors at night can achieve a major reduction in biting intensity by malaria vector mosquitoes. Persons sleeping at floor level bear a disproportionate risk of being bitten at night because this is the preferred height for feeding by the primary vector species. Therefore it is critical to protect children sleeping at floor level (bednets; repellent-impregnated blankets or sheets, etc.). Additionally, the opportunity exists for the development of inexpensive repellent-impregnated anklets and/or sandals to discourage vectors feeding on the lower legs under outdoor conditions at night.

Hemozoin activates the innate immune system and reduces Plasmodium berghei infection in Anopheles gambiae.

BackgroundMalaria is a worldwide infectious disease caused by Plasmodium parasites and transmitted by female Anopheles mosquitoes. The malaria vector mosquito Anopheles can trigger effective mechanisms to control completion of the Plasmodium lifecycle; the mosquito immune response to the parasite involves several pathways which are not yet well characterized. Plasmodium metabolite hemozoin has emerged as a potent immunostimulator of mammalian tissues. In this study, we aim to investigate the role of this parasite’s by-product as stimulator of Anopheles gambiae immunity to Plasmodium berghei.MethodsFemale mosquitoes were inoculated with hemozoin and the Plasmodium infection rate and intensity were measured. Differences between treatments were detected by Zero-inflated models. Microarray transcription analysis was performed to assess gene expression response to hemozoin. Genome-wide analysis results were confirmed by stimulation of Anopheles gambiae tissues and cells with hemozoin and silencing of REL2-F and its negative regulator Caspar.ResultsGene expression profiles revealed that hemozoin activates several immunity genes, including pattern recognition receptors (PRRs) and antimicrobial peptides (AMPs). Importantly, we found that the Immune deficiency (Imd) pathway Nuclear Factor-kappaB (NF-κB) transcription factor REL2, in its full-length form REL2-F, was induced upon hemozoin treatment.ConclusionsWe have for the first time shown the impact of hemozoin treatment in Plasmodium infection, reducing both rate and intensity of the infection. We propose that hemozoin boosts the innate immunity in Anopheles, activating key effector genes involved in mosquito resistance to Plasmodium, and this activation is REL2-mediated.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-014-0619-y) contains supplementary material, which is available to authorized users.

ケニア西部（GembeEast,Mbita）におけるマラリア媒介蚊のピレスロイド感受性に関する調査（４）行動抵抗性の有無に関する検討

ケニア西部（GembeEast,Mbita）におけるマラリア媒介蚊のピレスロイド感受性に関する調査（４）行動抵抗性の有無に関する検討

ケニア西部(Gembe East,Mbita)におけるマラリア媒介蚊のピレスロイド感受性に関する調査(5)ハマダラカのピレスロイドに対する忌避行動に関する検討

ケニア西部(Gembe East,Mbita)におけるマラリア媒介蚊のピレスロイド感受性に関する調査(5)ハマダラカのピレスロイドに対する忌避行動に関する検討