[Show abstract][Hide abstract] ABSTRACT: The malaria parasite-resistance island (PRI) of the African mosquito vector, Anopheles gambiae, was mapped to five genomic regions containing 80 genes, using coexpression patterns of genomic blocks. High-throughput sequencing identified 347 nonsynonymous single-nucleotide polymorphisms within these genes in mosquitoes from malaria-endemic areas in Kenya. Direct association studies between nonsynonymous single-nucleotide polymorphisms and Plasmodium falciparum infection identified three naturally occurring genetic variations in each of three genes (An. gambiae adenosine deaminase, fibrinogen-related protein 30, and fibrinogen-related protein 1) that were associated significantly with parasite infection. A role for these genes in the resistance phenotype was confirmed by RNA interference knockdown assays. Silencing fibrinogen-related protein 30 increased parasite infection significantly, whereas ablation of fibrinogen-related protein 1 transcripts resulted in mosquitoes nearly free of parasites. The discovered genes and single-nucleotide polymorphisms are anticipated to be useful in the development of tools for malaria control in endemic areas in Africa.
Full-text · Article · Dec 2013 · Proceedings of the National Academy of Sciences
[Show abstract][Hide abstract] ABSTRACT: Information on population age structure of mosquitoes under natural conditions is fundamental to the understanding of vectorial capacity and crucial for assessing the impact of vector control measures on malaria transmission. Transcriptional profiling has been proposed as a method for predicting mosquito age for Aedes and Anopheles mosquitoes, however, whether this new method is adequate for natural conditions is unknown. This study tests the applicability of transcriptional profiling for age-grading of Anopheles gambiae, the most important malaria vector in Africa. The transcript abundance of two An. gambiae genes, AGAP009551 and AGAP011615, was measured during aging under laboratory and field conditions in three mosquito strains. Age-dependent monotonic changes in transcript levels were observed in all strains evaluated. These genes were validated as age-grading biomarkers using the mark, release and recapture (MRR) method. The MRR method determined a good correspondence between actual and predicted age, and thus demonstrated the value of age classifications derived from the transcriptional profiling of these two genes. The technique was used to establish the age structure of mosquito populations from two malaria-endemic areas in western Kenya. The population age structure determined by the transcriptional profiling method was consistent with that based on mosquito parity. This study demonstrates that the transcription profiling method based on two genes is valuable for age determination of natural mosquitoes, providing a new approach for determining a key life history trait of malaria vectors.
[Show abstract][Hide abstract] ABSTRACT: Background & objectives:
The use of insecticides to eliminate mosquito larvae from ground pools may disrupt atural predator-induced control of mosquito larvae. Detrimental effects on predators may be directly from toxicity or by eliminating prey organisms. Identifying the principal predators responsible for mosquito suppression is needed to select non-target indicator species for insecticide studies. In this study, we sought to determine trophic level interactions between predators and immature stages of Anopheles gambiae Giles mosquitoes under experimental conditions in the coastal region of Kenya.
To identify effective predation pattern, a series of prey choice experiments was conducted. The relative abilities of five common species of aquatic insects found in the malaria-endemic coastal region of Kenya were assessed in a series of experiments. Experiments were conducted in semi-field conditions at Jaribuni, near the sites of insect collection.
In single predator experiments, notonectids consumed most of the mosquito larvae; hydrometrids consumed about half of the mosquito larvae in treatments. Veliids and gerrids had significant, but small effects on larval survivorship. Dytiscids did not have a significant effect on mosquito larvae survivorship. In a two-predator experiment, notonectids significantly decreased survivorship of dytiscids without a change in suppressive effects on mosquito larvae. Of the five common predators evaluated, notonectids were clearly the most voracious consumers of mosquito larvae. The predation pressure on mosquito larvae was not affected by the addition of additional prey items, consisting of small dytiscid beetles. The importance of this notonectid species in coastal Kenya suggests that it would be a valuable non-target indicator species for insecticide studies. Hydrometrids were also efficient at consuming mosquito larvae.
Interpretation & conclusion:
Of the five common predators from the Kenyan coast evaluated in this study, notonectids were the most voracious consumers of immature mosquitoes. Their predation pressure on mosquito larvae was not affected by the addition of additional prey items, consisting of small dytiscid beetles.
Full-text · Article · May 2013 · Journal of vector borne diseases
[Show abstract][Hide abstract] ABSTRACT: The mosquito midgut is the organ into which the blood meal passes and in which, within a peritrophic membrane secreted by the epithelium, the blood is retained during digestion and absorption. The mosquito midgut is lined with an actin filled microvilli that are exposed to the harsh environment of the gut lumen such as food particle abrasion, digestive hydrolases and attack by pathogens and parasites that are taken in by the blood meal. These microvilli are protected them these effects by the peritrophic matrix, the glycocalyx and the mucin proteins that line their epithelial surfaces. Immunization of BALB/c mice with AgMUC1/IL-12 cDNA has been shown to kill mosquitoes when fed on these mice. Mucin is one of the proteins produced in the mosquito midgut after a blood meal. The fine structure of the mosquito midgut epithelium interacting with immune factors such as antibodies or immune cells is of special significance for interpreting early events in the interaction between the mosquito midgut lining and the specific immune components present in the blood of AgMUC1/IL-12 cDNA immunized BALB/c mice. Following bright light microscopy, scanning electron and transmission electron microscopic observations of the features seen in mosquito midgut sections from An. gambiae mosquitoes fed on BALB/c mice immunized with AgMUC1/IL-12 cDNA, the most likely immune mechanisms responsible for mosquito killing could be cell mediated, most likely antibody dependent cellular cytotoxicity. Both necrotic and apoptotic processes that could be the cause of mosquito death were seen to take place in the cells lining the midgut epithelium.
No preview · Article · Mar 2013 · International Journal of Morphology
[Show abstract][Hide abstract] ABSTRACT: Background
Over the past 20 years, numerous studies have investigated the ecology and behaviour of malaria vectors and Plasmodium falciparum malaria transmission on the coast of Kenya. Substantial progress has been made to control vector populations and reduce high malaria prevalence and severe disease. The goal of this paper was to examine trends over the past 20 years in Anopheles species composition, density, blood-feeding behaviour, and P. falciparum sporozoite transmission along the coast of Kenya.
Using data collected from 1990 to 2010, vector density, species composition, blood-feeding patterns, and malaria transmission intensity was examined along the Kenyan coast. Mosquitoes were identified to species, based on morphological characteristics and DNA extracted from Anopheles gambiae for amplification. Using negative binomial generalized estimating equations, mosquito abundance over the period were modelled while adjusting for season. A multiple logistic regression model was used to analyse the sporozoite rates.
Results show that in some areas along the Kenyan coast, Anopheles arabiensis and Anopheles merus have replaced An. gambiae sensu stricto (s.s.) and Anopheles funestus as the major mosquito species. Further, there has been a shift from human to animal feeding for both An. gambiae sensu lato (s.l.) (99% to 16%) and An. funestus (100% to 3%), and P. falciparum sporozoite rates have significantly declined over the last 20 years, with the lowest sporozoite rates being observed in 2007 (0.19%) and 2008 (0.34%). There has been, on average, a significant reduction in the abundance of An. gambiae s.l. over the years (IRR = 0.94, 95% CI 0.90–0.98), with the density standing at low levels of an average 0.006 mosquitoes/house in the year 2010.
Reductions in the densities of the major malaria vectors and a shift from human to animal feeding have contributed to the decreased burden of malaria along the Kenyan coast. Vector species composition remains heterogeneous but in many areas An. arabiensis has replaced An. gambiae as the major malaria vector. This has important implications for malaria epidemiology and control given that this vector predominately rests and feeds on humans outdoors. Strategies for vector control need to continue focusing on tools for protecting residents inside houses but additionally employ outdoor control tools because these are essential for further reducing the levels of malaria transmission.
[Show abstract][Hide abstract] ABSTRACT: Carbon dioxide (CO(2)) present in exhaled air is the most important sensory cue for female blood-feeding mosquitoes, causing activation of long-distance host-seeking flight, navigation towards the vertebrate host and, in the case of Aedes aegypti, increased sensitivity to skin odours. The CO(2) detection machinery is therefore an ideal target to disrupt host seeking. Here we use electrophysiological assays to identify a volatile odorant that causes an unusual, ultra-prolonged activation of CO(2)-detecting neurons in three major disease-transmitting mosquitoes: Anopheles gambiae, Culex quinquefasciatus and A. aegypti. Importantly, ultra-prolonged activation of these neurons severely compromises their ability subsequently to detect CO(2) for several minutes. We also identify odours that strongly inhibit CO(2)-sensitive neurons as candidates for use in disruption of host-seeking behaviour, as well as an odour that evokes CO(2)-like activity and thus has potential use as a lure in trapping devices. Analysis of responses to panels of structurally related odours across the three mosquitoes and Drosophila, which have related CO(2)-receptor proteins, reveals a pattern of inhibition that is often conserved. We use video tracking in wind-tunnel experiments to demonstrate that the novel ultra-prolonged activators can completely disrupt CO(2)-mediated activation as well as source-finding behaviour in Aedes mosquitoes, even after the odour is no longer present. Lastly, semi-field studies demonstrate that use of ultra-prolonged activators disrupts CO(2)-mediated hut entry behaviour of Culex mosquitoes. The three classes of CO(2)-response-modifying odours offer powerful instruments for developing new generations of insect repellents and lures, which even in small quantities can interfere with the ability of mosquitoes to seek humans.
[Show abstract][Hide abstract] ABSTRACT: Supplemental figure 1: CO2 receptors are highly conserved in insects.
a, Alignment of the amino acid sequences of CO2 receptor orthologs using ClustalW from D. melanogaster (Dmel), A. gambiae (Agam), A.aegypti (Aaeg) and C. quinquefasciatus (Cpip). Recreated using sequences from Robertson et. al. 2009. b Mean spikes per second of cpA neuron for 3-sec stimulus of 0.15% CO2 overlayed with 0.5-sec odour, counted in 100-msec bins, n=5, error bars=s.e.m.
Supplemental figure 2: Volatility and solubility of CO2 response inhibitors
Percentage inhibition of the CO2 response as a function of a, vapor pressure, or b, solubility. Information obtained using Scifinder Scholar. Differing shapes indicate same compounds across different insects; A. gambiae (□), A. aegypti (▲), C. quinquefascitus (—), D. melanogaster (◊). Differing colors indicate chemical class; alcohol (red), aldehyde (orange), ester (pink), alkane (blue), ketone (green), acid (purple). n=3.
Supplemental figure 3: Butanone activates cpA neuron in mosquito and mimics CO2.
a, Representative trace from A. gambiae peg sensillum to a 1-sec stimulus of solvent (PO) or 2-butanone (10-1 dilution). b, Mean responses of the cpA neuron to 1-sec 2-butanone at indicated dilutions on A. gambiae (Ag), A. aegypti (Ae), and C. quinquefasciatus (Cx). n=5, error bars=s.e.m.
Supplemental Figure 4: Effect of individual odor mixture components on ultra-prolonged activation.
a, Mean responses of the cpA neuron to 1-sec 2,3-butanedione at indicated dilutions on A. gambiae (Ag), A. aegypti (Ae), and C. quinquefasciatus (Cx). n=5. error bars=s.e.m. b, Schematic of odour exposure sequence. c, Mean response of A. aegypti cpA neuron to a 1-sec CO2 stimulus after pre-exposure as indicated on X-axis with odorants at t=0 and t=15 sec. d, Representative traces (left) from peg sensillum and mean responses (right) from the cpA neurons to 0.15% and 0.33% CO2 after a pretreatment to 3-min of paraffin oil, or individual odor mixture components indicated. n=5, error bars=s.e.m.
Supplemental figure 5: Long-term Inhibition of CO 2 response in A. aegypti after pre-exposure to combinations of odorants.
Mean increase in frequency of response of the cpA neuron to stimulus of 1-sec 0.15% CO2 applied approx. every 30-sec, following a 3-sec pre-exposure to odor mixtures (10-2 dilution) or paraffin oil (PO). d4on=2,3-butanedione, 4al=butanal, 5al=pentanal, 6ol=hexanol. n=5, error bars=s.e.m. Spontaneous activity subtracted.
Supplemental figure 6: Behavior disruption in wind-tunnel
a, Scatterplot of time elapsed from start of assay required to find CO2 source for individual female Aedes mosquitoes according to the various ultra-prolonged blend treatments as indicated. Mosquitoes that did not find source are not included. Mean times and s.e.m. are indicated with line and error bars. b, Mean flight-time (after takeoff from holding cage) to the half-way point, and c, mean flight-time (after takeoff from holding cage) to the CO2 source. Error bars =s.e.m. Note the data for the 10-1 pre-exposure for 60-sec and 180-sec are severely skewed since very few individuals performed the behaviour, only 1 in the last condition. N=26 individuals for each condition were tested.
Supplemental Figure 7:
a,b Schematic diagram of the components of the stainless steel repellency chamber and use for the DEET avoidance assay.
Supplemental Figure 8:
Schematic and pictures of the a, experimental hut inside MalariaSphere with close-up pictures of the one-way entry traps installed in the eaves and TimeMist odour; b, remote mosquito release system; c, counter-flow geometry trap.
Supplemental Figure 9:
Pharmacophores of a, antagonists (1-hexanol, 1-pentanol, 1-butanal, 1-butanoic acid), and b, agonists (2-butanone and 2,3-butanedione). Pharmacophores are shown in two orientations at right-angles. Grey=hydrophobic, green=acceptors. Pharmacophores generated using PharmaGist program as described in Inbar, Y., et. al.. Deterministic Pharmacophore Detection via Multiple Flexible Alignment of Drug-Like Molecules. In Proc. of RECOMB (2007), vol. 3692 of Lecture Notes in Computer Science, pp. 423-434. Springer Verlag.
Supplemental Table 1:
a, Percentage of female C. quinquifasciatus mosquitoes trapped overnight in a counter-flow geometry trap using the indicated lures. N=5 trials per lure treatment, 50 mosquitoes/trial. b, Percentage of female C. quinquifasciatus mosquitoes flying through the odor emitting ring upwind in wind-tunnel assays. N= 20 per trial.
Supplemental Video 1:
Representative video of a mock-treated female A. aegypti mosquito navigating upwind and through a CO2 turbulent-plume releasing ring in real time.
Supplemental Video 2:
Representative video of a pretreated (ultra-prolonged odour blend, 10-2) female A. aegypti mosquito, unable to find upwind CO2 source in real time. For the remaining duration of the 300 sec assay the mosquito does not move after coming to rest on the glass wall.
Supplemental Video 3:
Representative video of a pretreated (ultra-prolonged odour blend, 10-1) female A. aegypti mosquito, unable to activate upwind flight from the cage and find upwind CO2 source during the entire duration of the 300 sec assay (video speeded up).
[Show abstract][Hide abstract] ABSTRACT: Larval control is a major component in mosquito control programs. This study evaluated the wide-scale application of Bti/Bs biolarvicide (Bacillus thuringiensis var. israelensis [Bti] and Bacillus sphaericus [Bs]) in different aquatic habitats in urban and peri-urban Malindi, Kenya. This study was done from June 2006 to December 2007. The urban and peri-urban area of Malindi town was mapped and categorized in grid cells of 1 km(2). A total of 16 1-km(2) cells were selected based on presence Community Based Organization dealing with malaria control within the cells. Each of the 16 1-km(2) cells was thoroughly searched for the presence of potential larval habitats. All habitats, whether positive or negative for larvae, were treated and rechecked 24 h (1 day), 6 days, and 10 days later for the efficacy of Bti/Bs. Weekly larval sampling was done to determine the mosquito larval dynamics in the aquatic habitats during the study period. Morphological identification of the mosquito larvae showed that Anopheles gambiae s.l. Giles was the most predominant species of the Anopheles and while in the culicines, Cx. quinquefasciatus Say was the predominant species. Anopheles larvae were all eliminated in habitats within a day post-application. For culicine larvae, 38.1% (n=8) of the habitat types responded within day 1 post-treatment and all the larvae were killed, they turned negative during the days of follow-up. Another 38.1% (n=8) of the habitat types had culicine larvae but turned negative by day 6, while three habitats (14.3%) had larvae by 6th day but turned negative by 10th day. However during this Bti/Bs application studies, two habitat types, house drainage and cesspits (9.5%), remained positive during the follow-up although the mosquito larvae were significantly reduced. Both early and late instars of Anopheles larvae immediately responded to Bti/Bs application and reached 100% mortality. The early and late instars of culicine responded to the Bti/Bs application but not as fast as the Anopheles larval instars. The early instars Culex, responded with 90.8% mortality at day 1 post-treatment, and the mortality was 99.9% at day 10. Similarly, the late instars Culex followed the same trend and exhibited same mortalities. The weekly sampling in the aquatic habitats showed that there was a 36.3% mosquito larval reduction in the aquatic habitats over the 18-months study period. In conclusion, Bti/Bs biolarvicide are useful in reducing the mosquito larval densities in a wide range of habitats which have a direct impact of adult mosquito populations.
Full-text · Article · Jun 2011 · Parasitology Research
[Show abstract][Hide abstract] ABSTRACT: Mosquitoes (Diptera: Culicidae) are important vectors of human disease-causing pathogens. Mosquitoes are found both in rural and urban areas. Deteriorating infrastructure, poor access to health, water and sanitation services, increasing population density, and widespread poverty contribute to conditions that modify the environment, which directly influences the risk of disease within the urban and peri-urban ecosystem. The objective of this study was to evaluate the mosquito vector abundance and diversity in urban, peri-urban, and rural strata in Malindi along the Kenya coast. The study was conducted in the coastal district of Malindi between January and December 2005. Three strata were selected which were described as urban, peri-urban, and rural. Sampling was done during the wet and dry seasons. Sampling in the wet season was done in the months of April and June to cover the long rainy season and in November and December to cover the short rainy season, while the dry season was between January and March and September and October. Adult mosquito collection was done using Pyrethrum Spray Collection (PSC) and Centers for Disease Control and Prevention (CDC) light traps inside houses and specimens were identified morphologically. In the three strata (urban, peri-urban, and rural), 78.5% of the total mosquito (n = 7,775) were collected using PSC while 18.1% (n = 1,795) were collected using the CDC light traps. Using oviposition traps, mosquito eggs were collected and reared in the insectary which yielded 329 adults of which 83.8% (n = 276) were Aedes aegypti and 16.2% (n = 53) were Culex quinquefasciatus. The mosquito distribution in the three sites varied significantly in each collection site. Anopheles gambiae, Anopheles funestus and Anopheles coustani were predominant in the rural stratum while C. quinquefasciatus was mostly found in urban and peri-urban strata. However, using PSC and CDC light trap collection techniques, A. aegypti was only found in urban strata. In the three strata, mosquitoes were mainly found in high numbers during the wet season. Further, A. gambiae, C. quinquefasciatus, and A. aegypti mosquitoes were found occurring together inside the houses. This in turn exposes the inhabitants to an array of mosquito-borne diseases including malaria, bancroftian filariasis, and arboviruses (dengue fever, Yellow fever, Rift Valley fever, Chikungunya fever, and West Nile Virus). In conclusion, our findings provide useful information for the design of integrated mosquito and disease control programs in East African environments.
Full-text · Article · May 2011 · Parasitology Research
[Show abstract][Hide abstract] ABSTRACT: The ecology of malaria vectors particularly in semi-arid areas of Africa is poorly understood. Accurate knowledge on this subject will boost current efforts to reduce the burden of malaria in sub-Saharan Africa. The objective of this study was to describe the dynamics of malaria transmission in two model semi-arid sites (Kamarimar and Tirion) in Baringo in Kenya.
Adult mosquitoes were collected indoors by pyrethrum spray collections (PSC) and outdoors by Centers for Disease Control (CDC) light traps and identified to species by morphological characteristics. Sibling species of Anopheles gambiae complex were further characterized by rDNA. PCR and enzyme-linked immuno-sorbent assays (ELISA) were used to test for Plasmodium falciparum circumsporozoite proteins and host blood meal sources respectively.
Anopheles arabiensis was not only the most dominant mosquito species in both study sites but also the only sibling species of An. gambiae s.l. present in the area. Other species identified in the study area were Anopheles funestus, Anopheles pharoensis and Anopheles coustani. For Kamarimar but not Tirion, the human blood index (HBI) for light trap samples was significantly higher than for PSC samples (Kamarimar, 0.63 and 0.11, Tirion, 0.48 and 0.43). The HBI for light trap samples was significantly higher in Kamarimar than in Tirion while that of PSC samples was significantly higher in Tirion than in Kamarimar. Entomological inoculation rates (EIR) were only detected for one month in Kamarimar and 3 months in Tirion. The number of houses in a homestead, number of people sleeping in the house, quality of the house, presence or absence of domestic animals, and distance to the animal shelter and the nearest larval habitat were significant predictors of An. arabiensis occurrence.
Malaria transmission in the study area is seasonal with An. arabiensis as the dominant vector. The fact this species feeds readily on humans and domestic animals suggest that zooprophylaxis may be a plausible malaria control strategy in semi-arid areas of Africa. The results also suggest that certain household characteristics may increase the risk of malaria transmission.
[Show abstract][Hide abstract] ABSTRACT: More focus is given to mosquito larval control due to the necessity to use several control techniques together in integrated vector management programmes. Botanical products are thought to be able to provide effective, sustainable and cheap mosquito larval control tools. However, bio-larvicides like Azadirachta indica (neem) could repel adult mosquitoes from laying their eggs in the treated larval habitats. In this study the response of Anopheles gambiae s.s. mosquitoes towards varying doses of crude aqueous neem extracts was examined. Non-choice oviposition tests were used to measure the proportion of mosquitoes laying on the first or second night, or not laying at all, when compared to the control. For each individual mosquito, the number of eggs laid and/or retained in the ovary was counted to determine the relationship between wing length and egg production. Larger female mosquitoes produced larger egg batches. The results show that at a dose of 0.1 g/l, a concentration previously found to be effective at controlling mosquito larvae, the oviposition behaviour of adult female mosquitoes was not significantly affected. The results indicate that the mosquitoes would expose progeny to this neem control tool, making the use of these simple neem wood extracts effective and potentially sustainable.
Full-text · Article · Apr 2011 · Journal of medicinal plant research
[Show abstract][Hide abstract] ABSTRACT: Pre-adult stages of malaria vectors in semi-arid areas are confronted with highly variable and challenging climatic conditions. The objective of this study was to determine which larval habitat types are most productive in terms of larval densities in the dry and wet seasons within semi-arid environments, and how vector species productivity is partitioned over time.
Larval habitats were mapped and larvae sampled longitudinally using standard dipping techniques. Larvae were identified to species level morphologically using taxonomic keys and to sub-species by polymerase chain reaction (PCR) methods. Physical characteristics of larval habitats, including water depth, turbidity, and presence of floating and emergent vegetation were recorded. Water depth was measured using a metal ruler. Turbidity, pH, conductivity, dissolved oxygen, temperatures salinity and total dissolved solids (TDS) were measured in the field using the hand-held water chemistry meters.
Mean larval densities were higher in the dry season than during the wet season but the differences in density were not statistically significant (F = 0.04, df = 1, p = 0.8501). Significantly higher densities of larvae were collected in habitats that were shaded and holding turbid, temporary and still water. Presence of emergent or floating vegetation, habitat depth, habitat size and habitat distance to the nearest house did not significantly affect larval density in both villages. There was a weakly positive relationship between larval density and salinity (r = 0.19, p < 0.05), conductivity (r = 0.05, p = 0.45) and total dissolved solids (r = 0.17, p < 0.05). However, the relationship between water temperature and larval density was weakly negative (r = 0.15, p = 0.35). All statistical tests were significant at alpha = 0.05.
Breeding of malaria vector mosquitoes in Baringo is driven by predominantly human-made and permanent breeding sites in which Anopheles arabiensis and Anopheles funestus breed at a low level throughout the year. Permanent water sources available during the dry season serve as inocula by providing "larval seed" to freshly formed rain-fed habitats during the rainy season. The highly localized and focal nature of breeding sites in these semi-desert environments provides a good opportunity for targeted larval control since the habitats are few, well-defined and easily traceable.
Full-text · Article · Feb 2011 · Parasites & Vectors