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Sample records for anopheles gambiae diptera

  1. The spatial distribution of Anopheles gambiae sensu stricto and An. arabiensis (Diptera: Culicidae in Mali

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    N. Sogoba

    2007-05-01

    Full Text Available Variations in the biology and ecology and the high level of genetic polymorphism of malaria vectors in Africa highlight the value of mapping their spatial distribution to enhance successful implementation of integrated vector management. The objective of this study was to collate data on the relative frequencies of Anopheles gambiae s.s. and An. arabiensis mosquitoes in Mali, to assess their association with climate and environmental covariates, and to produce maps of their spatial distribution. Bayesian geostatistical logistic regression models were fitted to identify environmental determinants of the relative frequencies of An. gambiae s.s. and An. arabiensis species and to produce smooth maps of their geographical distribution. The frequency of An. arabiensis was positively associated with the normalized difference vegetation index, the soil water storage index, the maximum temperature and the distance to water bodies. It was negatively associated with the minimum temperature and rainfall. The predicted map suggests that, in West Africa, An. arabiensis is concentrated in the drier savannah areas, while An. gambiae s.s. prefers the southern savannah and land along the rivers, particularly the inner delta of Niger. Because the insecticide knockdown resistance (kdr gene is reported only in An. gambiae s.s. in Mali, the maps provide valuable information for vector control. They may also be useful for planning future implementation of malaria control by genetically manipulated mosquitoes.

  2. Unassisted isolated-pair mating of Anopheles gambiae (Diptera: Culicidae) mosquitoes.

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    Benedict, Mark Q; Rafferty, Cristina S

    2002-11-01

    Female Anopheles mosquitoes usually mate only once, but mating is seldom seen in small containers containing only one female and male. Therefore, matings are often performed among many adults in large cages or by forced copulation. Isolated-pair mating of Anopheles gambiae G3 strain-derived mosquitoes without forced copulation in small vials is described. We observed that the experimental variables eye color and male number were significant factors in the mating frequency. Females mated more frequently when three males were present over only one male. White-eyed females were more likely to be mated than wild-eyed females, but wild males mated more frequently than did white-eyed males. Experiments were also conducted to determine when mating was occurring by using wild-eye-color mosquitoes in isolated pairs. Almost no matings were observed before day 6 rather than the frequencies typically observed after 1-2 d in standard large-cage matings among large numbers of adults.

  3. First report of Metarhizium anisopliae IP 46 pathogenicity in adult Anopheles gambiae s.s. and An. arabiensis (Diptera; Culicidae)

    NARCIS (Netherlands)

    Mnyone, L.L.; Russell, T.L.; Lyimo, I.N.; Lwetoijera, D.W.; Kirby, M.J.; Luz, C.

    2009-01-01

    The entomopathogenic fungus Metarhizium anisopliae isolate IP 46, originating from a soil sample collected in 2001 in the Cerrado of Central Brazil, was tested for its ability to reduce the survival of adult male and female Anopheles gambiae s.s. and An. arabiensis mosquitoes. A 6-h exposure to the

  4. Extent of digestion affects the success of amplifying human DNA from blood meals of Anopheles gambiae (Diptera: Culicidae)

    NARCIS (Netherlands)

    Mukabana, W.R.; Takken, W.; Seda, P.; Killeen, G.F.; Hawley, W.A.; Knols, B.G.J.

    2002-01-01

    The success of distinguishing blood meal sources of Anopheles gambiae Giles through deoxyribonucleic acid (DNA) profiling was investigated by polymerase chain reaction (PCR) amplification at the TC-11 and VWA human short tandem repeats (STR) loci. Blood meal size and locus had no significant effect

  5. Predators of Anopheles gambiae sensu lato (Diptera: Culicidae) larvae in wetlands, western Kenya: confirmation by polymerase chain reaction method.

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    Ohba, Shin-Ya; Kawada, Hitoshi; Dida, Gabriel O; Juma, Duncan; Sonye, Gorge; Minakawa, Noboru; Takagi, Masahiro

    2010-09-01

    Polymerase chain reaction analysis was performed to determine whether mosquito predators in wetland habitats feed on Anopheles gambiae sensu lato (s.l.) larvae. Aquatic mosquito predators were collected from six wetlands near Lake Victoria in Mbita, Western Kenya. This study revealed that the whole positive rate of An. gambiae s.l. from 330 predators was 54.2%. The order of positive rate was the highest in Odonata (70.2%), followed by Hemiptera (62.8%), Amphibia (41.7%), and Coleoptera (18%). This study demonstrates that the polymerase chain reaction method can determine whether aquatic mosquito predators feed on An. gambiae s.l. larvae if the predators have undigested An. gambiae s.l. in their midgut or stomach. PMID:20939371

  6. Cannibalism and predation among larvae of the Anopheles gambiae complex

    NARCIS (Netherlands)

    Koenraadt, C.J.M.; Takken, W.

    2003-01-01

    Among the aquatic developmental stages of the Anopheles gambiae complex (Diptera: Culicidae), both inter- and intra-specific interactions influence the resulting densities of adult mosquito populations. For three members of the complex, An. arabiensis Patton, An. quadriannulatus (Theobald) and An. g

  7. Predators of Anopheles gambiae sensu lato (Diptera: Culicidae) Larvae in Wetlands, Western Kenya: Confirmation by Polymerase Chain Reaction Method

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    Ohba, Shin-ya; Kawada, Hitoshi; Dida, Gabriel O; Juma, Duncan; SONYE, GORGE; Minakawa, Noboru; Takagi, Masahiro

    2010-01-01

    Polymerase chain reaction analysis was performed to determine whether mosquito predators in wetland habitats feed on Anopheles gambiae sensu lato (s.l.) larvae. Aquatic mosquito predators were collected from six wetlands near Lake Victoria in Mbita, Western Kenya. This study revealed that the whole positive rate of An. gambiae s.l. from 330 predators was 54.2%. The order of positive rate was the highest in Odonata (70.2%), followed by Hemiptera (62.8%), Amphibia (41.7%), and Coleoptera (18%)....

  8. Predators of Anopheles gambiae sensu lato (Diptera: Culicidae) larvae in Wetlands, Western Kenya: Confirmation by polymerase chain reaction method

    OpenAIRE

    Ohba, Shin-ya; Kawada, Hitoshi; Dida, Gabriel O; Juma, Duncan; SONYE, GORGE; Minakawa, Noboru; Takagi, Masahiro

    2010-01-01

    Polymerase chain reaction analysis was performed to determine whether mosquito predators in wetland habitats feed on Anopheles gambiae sensu lato (s.l.) larvae. Aquatic mosquito predators were collected from six wetlands near Lake Victoria in Mbita, Western Kenya. This study revealed that the whole positive rate of An. gambiae s.l. from 330 predators was 54.2%. The order of positive rate was the highest in Odonata (70.2%), followed by Hemiptera (62.8%), Amphibia (41.7%), and Coleoptera (18%)....

  9. Effect of larval environment on some life history parameters in anopheles gambiae s.s. (diptera:culicidae))

    OpenAIRE

    Jannat, Khandaker Noore

    2010-01-01

    The effects of larval density, nutrition and cannibalism risk on some life history parameters of Anopheles gambiae larvae were evaluated in the laboratory. Adult body size was inversely correlated with larval density whereas larval mortality and mean age at pupation varied across experiments. When density increased, the secondary sex ratio shifted toward female bias. Effects of different types of nutrition on larval life were compared by providing larvae with algae Chaetophora sp., fish food ...

  10. Identification, characterisation, and function of adipokinetic hormones and receptor in the African malaria mosquito, "Anopheles Gambiae" (Diptera)

    OpenAIRE

    Kaufmann, Christian; Betschart, Bruno

    2007-01-01

    En utilisant la bioinformatique et la biologie moléculaire, nous avons pu identifier chez le principal vecteur africain de la malaria, le moustique, Anopheles gambiae deux hormones adipokinétiques (AKHs): l'octapeptide, Anoga-AKH-I (pQLTFTPAWa) et le décapeptide, Anoga-AKH-II, (pQVTFSRDWNAa). La fonction principale des AKHs est d’induire une hyperlipémie (effet d’adipokinétique), ainsi qu’une hypertrehalosémie et une hyperprolinémie. En tant que membres de la famille des AKH, les deux neurope...

  11. Repellent activities of stereoisomers of p-menthane-3,8-diols against Anopheles gambiae (Diptera: Culicidae).

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    Barasa, Stephen S; Ndiege, Isaiah O; Lwande, Wilber; Hassanali, Ahmed

    2002-09-01

    Four stereoisomers of p-menthane-3,8-diol, which make up the natural product obtained from Eucalyptus citriodora, were synthesized through stereoselective procedures. Repellency assays showed that all the four were equally active against Anopheles gambiae s.s. Racemic blends and the diastereoisomeric mixture of all the four isomers were also equally repellent. 1-alpha-terpeneol, with a single hydroxyl function at C-8 and unsaturation at C-8, and menthol, with a single hydroxyl function at C-3, were not repellent. The practical implication of these results is discussed. PMID:12349856

  12. Comparison of the Insecticidal Characteristics of Commercially Available Plant Essential Oils Against Aedes aegypti and Anopheles gambiae (Diptera: Culicidae).

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    Norris, Edmund J; Gross, Aaron D; Dunphy, Brendan M; Bessette, Steven; Bartholomay, Lyric; Coats, Joel R

    2015-09-01

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

  13. Orientation of Anopheles gambiae (Diptera: Culicidae) to Plant-Host Volatiles in a Novel Diffusion-Cage Olfactometer.

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    Otienoburu, Philip E; Nikbakhtzadeh, Mahmood R; Foster, Woodbridge A

    2016-01-01

    A novel diffusion-cage olfactometer tested the responses of Anopheles gambiae Giles to plant volatiles. Green-leaf volatiles are often released from cut or injured plant tissue and may alter the headspace of plants used in olfactometer assays. The diffusion-cage olfactometer is designed for use with whole, intact plants, hence giving a more realistic behavioral assay. Its simple plastic construction, ease of assembly, and accommodation to whole plants makes it a useful tool for measuring mosquito orientation to plant volatiles within large enclosures. We compared its performance to that of the more commonly used T-tube wind-tunnel olfactometer, by testing the orientation of mosquitoes to volatiles of a few prevalent plants of eastern Africa reportedly utilized by An. gambiae for sugar: Parthenium hysterophorus (Asteraceae), Ricinus communis (Euphorbiaceae), Lantana camara (Verbenaceae), and Senna occidentalis (Fabaceae). Results indicate that the diffusion-cage olfactometer is an effective alternative to conventional wind-tunnel olfactometers, to test mosquito orientation to plant volatiles under seminatural conditions. PMID:26502752

  14. Toxicity of pirimiphos methyl (Actellic 25EC) on Anopheles gambiae s.s., Culex quinquefasciatus (Diptera: Culicidae), and potential biocontrol agent, Poecilia reticulata (Pisces: Poeciliidae).

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    Anogwih, Joy A

    2014-08-01

    The toxicity of an emulsifiable formulation of pirimiphos methyl (Actellic 25EC) on Anopheles gambiae s.s. Giles, Culex quinquefasciatus Say (Diptera: Culicidae), and predator fish Poecilia reticulata Peters (Pisces: Poeciliidae) was investigated. Acute toxicity tests were carried out to determine the effect of the larvicide on mosquito larvae and fish species. To investigate the nontarget effects on P. reticulata, fish of similar size (3.5 +/- 0.2 cm) were randomly selected and exposed for 28 d, under static renewal bioassay, to sublethal concentrations of the larvicide capable of killing 30 and 70% of Cx. quinquefasciatus. The 24 h LC50 value of pirimiphos methyl on the test organisms ranged between 20.44 and 697.30 microg liter(-1). The ultrastructural changes observed in the intestinal cells of P. reticulata were characterized by degenerating cell membranes with gradual loss of gray area in pycnotic nucleus at lower concentration. Marked damage was found at higher concentration including distinct loss of gray areas in cytosol, absence of cristae, numerous ruptures, and several dead cells. Pirimiphos methyl was toxic to a predatory fish species, and for its relevance in vector control and crop protection, warrants cumulative assessment to establish its comprehensive ecological risk, and the dosage required for field larviciding.

  15. Synergism between ammonia, lactic acid and carboxylic acids as kairomones in the host-seeking behaviour of the malaria mosquito Anopheles gambiae sensu stricto (Diptera: Culicidae)

    NARCIS (Netherlands)

    Smallegange, R.C.; Qiu, Y.T.; Loon, van J.J.A.; Takken, W.

    2005-01-01

    Host odours play a major role in the orientation and host location of blood-feeding mosquitoes. Anopheles gambiae Giles sensu stricto, which is the most important malaria vector in Africa, is a highly anthropophilic mosquito species, and the host-seeking behaviour of the females of this mosquito is

  16. Neuropeptides and Peptide Hormones in Anopheles gambiae

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    Riehle, Michael A.; Garczynski, Stephen F.; Crim, Joe W.; Hill, Catherine A.; Brown, Mark R.

    2002-10-01

    The African malaria mosquito, Anopheles gambiae, is specialized for rapid completion of development and reproduction. A vertebrate blood meal is required for egg production, and multiple feedings subsequently allow transmission of malaria parasites, Plasmodium spp. Regulatory peptides from 35 genes annotated from the A. gambiae genome likely coordinate these and other physiological processes. Plasmodium parasites may affect actions of newly identified insulin-like peptides, which coordinate growth and reproduction of its vector, A. gambiae, as in Drosophila melanogaster, Caenorhabditis elegans, and mammals. This genomic information provides a basis to expand understanding of hematophagy and pathogen transmission in this mosquito.

  17. Male motion coordination in swarming Anopheles gambiae and Anopheles coluzzii

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    The Anopheles gambiae species complex comprises the primary vectors of malaria in much of sub-Saharan Africa; most of the mating in these species occurs in swarms composed almost entirely of males. Intermittent, parallel flight patterns in such swarms have been observed, but a detailed description o...

  18. Preliminary field testing of a long-lasting insecticide-treated hammock against Anopheles gambiae and Mansonia spp. (Diptera : Culicidae) in West Africa

    OpenAIRE

    Hougard, Jean-Marc; Martin, Thibaud; Guillet, Pierre; Coosemans, M.; ITOH, T.; Akogbéto, M.; Chandre, Fabrice

    2007-01-01

    The efficacy of an experimental long-lasting insecticide-treated ham mock (LLIH) with a long-lasting treated net used as a blanket and made of the same fabric (polyethylene) was tested in a concrete block experimental hut, against the malaria vector Anopheles gambiae s.l. and the arbovirus vectors and nuisance mosquitoes Mansonia africana (Theobald) and Alansonia uniformis (Theobald). The LLIH was treated with the pyrethroid insecticide permethrin. It was evaluated concurrently with ignited m...

  19. Energy-state dependent responses of Anopheles gambiae to an unobtainable host

    OpenAIRE

    Zappia, Simon Pierre William

    2011-01-01

    Understanding how blood-seeking behavior changes with different energy levels in the malaria mosquito Anopheles gambiae (Diptera: Culicidae), when confronted with an unobtainable blood-host, is of interest for vector control strategies. I used a straight-tube olfactometer to mimic a domicile containing (i) a simulated blood-host (human foot smell) protected by either a plain bednet or a DEET impregnated net and (ii) a sugar source (honey scent) some distance away. I manipulated the mosquito’s...

  20. Transstadial and horizontal transfer of bacteria within a colony of Anopheles gambiae (Diptera: Culicidae) and oviposition response to bacteria-containing water.

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    Lindh, J M; Borg-Karlson, A-K; Faye, I

    2008-09-01

    In a paratransgenic approach, genetically modified bacteria are utilized to kill the parasite in the vector gut. A critical component for paratransgenics against malaria is how transgenic bacteria can be introduced and then kept in a mosquito population. Here, we investigated transstadial and horizontal transfer of bacteria within an Anopheles gambiae mosquito colony with the focus on spiked breeding sites as a possible means of introducing bacteria to mosquitoes. A Pantoea stewartii strain, previously isolated from An. gambiae, marked with a green fluorescent protein (GFP), was introduced to mosquitoes in different life stages. The following life stages or older mosquitoes in the case of adults were screened for bacteria in their guts. In addition to P. stewartii other bacteria were isolated from the guts: these were identified by 16S rRNA sequence analysis and temporal temperature gradient gel electrophoresis (TTGE). Bacteria were transferred from larvae to pupae but not from pupae to adults. The mosquitoes were able to take up bacteria from the water they emerged from and transfer the same bacteria to the water they laid eggs in. Elizabethkingia meningoseptica was more often isolated from adult mosquitoes than P. stewartii. A bioassay was used to examine An. gambiae oviposition responses towards bacteria-containing solutions. The volatiles emitted from the solutions were sampled by headspace-solid phase microextraction (SPME) and identified by gas chromatography and mass spectrometry (GC-MS) analysis. P. stewartii but not E. meningoseptica mediated a positive oviposition response. The volatiles emitted by P. stewartii include indole and 3-methyl-1-butanol, which previously have been shown to affect An. gambiae mosquito behaviour. E. meningoseptica emitted indole but not 3-methyl-1-butanol, when suspended in saline. Taken together, this indicates that it may be possible to create attractive breeding sites for distribution of genetically modified bacteria in the

  1. Comparative biology and reproductive behaviour of a laboratory-adapted Redco strain of Anopheles Gambiae Giles (Diptera; culicidae and wild populations of the same species

    International Nuclear Information System (INIS)

    The sterile insect technique involves mass rearing of male insects for sterility purpose. This heavily relies on male fitness and genetic compatibility of laboratory-adapted male insects and the wild to ensure successful competition with their male counterpart in the wild. Uniform environment in the laboratory as compared to the wild conditions might lead to genetic drift which might lead to reduced sexual competitiveness, fitness, morphological changes or changes in the sexual behaviour of mosquitoes. This work investigated the sexual compatibility, morphometry and sexual behaviour of laboratory-adapted strain and wild strain of Anopheles gambiae under laboratory conditions. These measurements were done by observing swarm formation, genitalia rotation, percentage insemination, fecundity, fertility, wing length, wing width, thoracic width, body length, body size index and wing size index. Morphometric studies of laboratory-adapted and wild strain of Anopheles gambiae were carried out by observing the wing length, body length and thoracic length under Lecia 4D stereoscope in order to find out variations in the body size between the two strains. The results showed significant difference between thoracic width and wing length between the laboratory-adapted strain and wild strain. Indices such as body size index and wing length index also showed significant difference between the two strains; laboratory-adapted REDCO strain (BSI 4.45 ± 0.10, p = 0.010 ; WSI 1.92 ± 0.07, p = 0.026) and wild REDCO strain ( 4.08 ± 0.10, p = 0.010 ; WSI 1.73 ± 0.04, p = 0.026 ). Body length of laboratory-adapted male mosquitoes (4.24 ± 0.05, p = 0.462) was not significantly different from its thoracic width, wing length, and wing width. The wild strain on the other hand had significant difference between its body length (4.19 ± 0.04, p = 0.462), thoracic width (0.096 ± 0.02, p = 0.002 ) and wing length (2.99 ± 0.03, p = 0.050 ). In the mating experiment, egg production in each of

  2. Genomic islands of speciation in Anopheles gambiae.

    Directory of Open Access Journals (Sweden)

    2005-09-01

    Full Text Available The African malaria mosquito, Anopheles gambiae sensu stricto (A. gambiae, provides a unique opportunity to study the evolution of reproductive isolation because it is divided into two sympatric, partially isolated subtaxa known as M form and S form. With the annotated genome of this species now available, high-throughput techniques can be applied to locate and characterize the genomic regions contributing to reproductive isolation. In order to quantify patterns of differentiation within A. gambiae, we hybridized population samples of genomic DNA from each form to Affymetrix GeneChip microarrays. We found that three regions, together encompassing less than 2.8 Mb, are the only locations where the M and S forms are significantly differentiated. Two of these regions are adjacent to centromeres, on Chromosomes 2L and X, and contain 50 and 12 predicted genes, respectively. Sequenced loci in these regions contain fixed differences between forms and no shared polymorphisms, while no fixed differences were found at nearby control loci. The third region, on Chromosome 2R, contains only five predicted genes; fixed differences in this region were also verified by direct sequencing. These "speciation islands" remain differentiated despite considerable gene flow, and are therefore expected to contain the genes responsible for reproductive isolation. Much effort has recently been applied to locating the genes and genetic changes responsible for reproductive isolation between species. Though much can be inferred about speciation by studying taxa that have diverged for millions of years, studying differentiation between taxa that are in the early stages of isolation will lead to a clearer view of the number and size of regions involved in the genetics of speciation. Despite appreciable levels of gene flow between the M and S forms of A. gambiae, we were able to isolate three small regions of differentiation where genes responsible for ecological and behavioral

  3. Physiology and development of the M and S molecular forms of Anopheles gambiae in Burkina Faso (West Africa)

    OpenAIRE

    Mouline, Karine; Mamai, W.; Agnew, P.; Tchonfienet, M.; Brengues, Cécile; Dabiré, R.; Robert, Vincent; Simard, Frédéric

    2012-01-01

    In West Africa, M and S molecular forms of Anopheles gambiae sensu stricto (Diptera: Culicidae) Giles, frequently occur together, although with different population bionomics. The S form typically breeds in rain-dependant water collections and is present during the rainy season only whereas the M form can thrive all year long in areas with permanent breeding opportunities. In the present study, we explored physiological and developmental trade-offs at play in laboratory colonies and field pop...

  4. CLIP proteases and Plasmodium melanization in Anopheles gambiae.

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    Barillas-Mury, Carolina

    2007-07-01

    Melanization is a potent immune response mediated by phenoloxidase (PO). Multiple Clip-domain serine proteases (CLIP) regulate PO activation as part of a complex cascade of proteases that are cleaved sequentially. The role of several CLIP as key activators or suppressors of the melanization responses of Anopheles gambiae to Plasmodium berghei (murine malaria) has been established recently using a genome-wide reverse genetics approach. Important differences in regulation of PO activation between An. gambiae strains were also identified. This review summarizes these findings and discusses our current understanding of the An. gambiae melanization responses to Plasmodium. PMID:17512801

  5. Identification of candidate volatiles that affect the behavioural response of the malaria mosquito Anopheles gambiae sensu stricto to an active kairomone blend: laboratory and semi-field assays

    NARCIS (Netherlands)

    Smallegange, R.C.; Bukovinszkine Kiss, G.; Otieno, B.; Mbadi, P.A.; Takken, W.; Mukabana, W.R.; Loon, van J.J.A.

    2012-01-01

    Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is the most important vector of human malaria in sub-Saharan Africa, affecting the lives of millions of people. Existing tools such as insecticide-treated nets and indoor-residual sprays are not only effective, but also have limitations as a

  6. Structure of host-odour plumes influences catch of Anopheles gambiae s.s. and Aedes aegypti in a dual-choice olfactometer

    NARCIS (Netherlands)

    Dekker, T.; Takken, W.; Cardé, R.T.

    2001-01-01

    The effects of structure, concentration and composition of host-odour plumes on catch of female Anopheles gambiae Giles sensu stricto and Aedes aegypti (L.) (Diptera: Culicidae) were investigated in a dual-choice olfactometer. We demonstrate that the fine-scale structure of host-odour plumes modulat

  7. Insecticide resistance status in Anopheles gambiae in southern Benin

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    Corbel Vincent

    2010-03-01

    Full Text Available Abstract Background The emergence of pyrethroid resistance in Anopheles gambiae has become a serious concern to the future success of malaria control. In Benin, the National Malaria Control Programme has recently planned to scaling up long-lasting insecticidal nets (LLINs and indoor residual spraying (IRS for malaria prevention. It is, therefore, crucial to monitor the level and type of insecticide resistance in An. gambiae, particularly in southern Benin where reduced efficacy of insecticide-treated nets (ITNs and IRS has previously been reported. Methods The protocol was based on mosquito collection during both dry and rainy seasons across forty districts selected in southern Benin. Bioassay were performed on adults collected from the field to assess the susceptibility of malaria vectors to insecticide-impregnated papers (permethrin 0.75%, delthamethrin 0.05%, DDT 4%, and bendiocarb 0.1% following WHOPES guidelines. The species within An. gambiae complex, molecular form and presence of kdr and ace-1 mutations were determined by PCR. Results Strong resistance to permethrin and DDT was found in An. gambiae populations from southern Benin, except in Aglangandan where mosquitoes were fully susceptible (mortality 100% to all insecticides tested. PCR showed the presence of two sub-species of An. gambiae, namely An. gambiae s.s, and Anopheles melas, with a predominance for An. gambiae s.s (98%. The molecular M form of An. gambiae was predominant in southern Benin (97%. The kdr mutation was detected in all districts at various frequency (1% to 95% whereas the Ace-1 mutation was found at a very low frequency (≤ 5%. Conclusion This study showed a widespread resistance to permethrin in An. gambiae populations from southern Benin, with a significant increase of kdr frequency compared to what was observed previously in Benin. The low frequency of Ace-1 recorded in all populations is encouraging for the use of bendiocarb as an alternative insecticide to

  8. Shift in species composition in the Anopheles gambiae complex after implementation of long-lasting insecticidal nets in Dielmo, Senegal.

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    Sougoufara, S; Harry, M; Doucouré, S; Sembène, P M; Sokhna, C

    2016-09-01

    Long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) are the cornerstones of malaria vector control. However, the effectiveness of these control tools depends on vector ecology and behaviour, which also largely determine the efficacy of certain Anopheles mosquitoes (Diptera: Culicidae) as vectors. Malaria vectors in sub-Saharan Africa are primarily species of the Anopheles gambiae complex, which present intraspecific differences in behaviour that affect how they respond to vector control tools. The focus of this study is the change in species composition in the An. gambiae complex after the implementation of LLINs in Dielmo, Senegal. The main findings referred to dramatic decreases in the proportions of Anopheles coluzzii and An. gambiae after the introduction of LLINs, and an increase in the proportion of Anopheles arabiensis. Two years after LLINs were first introduced, An. arabiensis remained the most prevalent species and An. gambiae had begun to rebound. This indicated a need to develop additional vector control tools that can target the full range of malaria vectors. PMID:27058993

  9. Studies of Anopheles gambiae s.l (Diptera: Culicidae exhibiting different vectorial capacities in lymphatic filariasis transmission in the Gomoa district, Ghana

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    Amuzu Hilaria

    2010-09-01

    Full Text Available Abstract Background Two lymphatic filariasis endemic communities Mampong and Hwida in Ghana have been regularly monitored for impact on transmission after annual mass drug administration (MDA with albendazole and ivermectin. After six MDAs even though the ABR for Mampong was 55883/person/year and that of Hwida was 2494/person/year, they both had ATPs of 15.21 infective larvae/person/year. Interestingly the human microfilaraemia levels had reduced significantly from 14% to 0% at Mampong and 12% to 3% at Hwida. In an attempt to understand this anomaly, we collected mosquitoes over a 5-month period using human landing catches to determine the species composition, the number of cibarial teeth, the lengths and widths of the cibarium and the cibarial dome of the vector populations. Results Out of 2553 mosquitoes caught at Mampong, 42.6% were An. gambiae s.l. All 280 identified further by PCR were An. gambiae s.s (275 M and 5 S molecular forms. At Hwida, 112 mosquitoes were obtained; 67 (59.8% were An. gambiae s.l, comprised of 40 (59.7% An. melas, 24 (35.8% An. gambiae s.s (17 and 5 M and S molecular forms respectively and 3 (4.5% unidentified. The mean number of teeth for An. melas was 14.1 (median = 14, range = 12-15, An. gambiae s.s., 15.7 (median = 15, range = 13-19 M form 15.5 (median = 15 range = 13-19 and S form 16 (median = 16, range 15-17. The observed differences in teeth numbers were significantly different between An. melas and An. gambiae s.s (p = 0.004, and the M form (p = 0.032 and the S form (p = 0.002. Conclusions In this study, An. gambiae s.s was the main vector at Mampong and was found to possess significantly more cibarial teeth than An. melas, the principal vector at Hwida. We postulate that the different impact observed after 6 MDAs may be due to An. gambiae s.s exhibiting 'facilitation' at Mampong and at Hwida An. melas the main vector exhibits 'limitation'. Thus it may be necessary to compliment MDA with vector control to

  10. The nicotinic acetylcholine receptor gene family of the malaria mosquito, Anopheles gambiae.

    Science.gov (United States)

    Jones, Andrew K; Grauso, Marta; Sattelle, David B

    2005-02-01

    Nicotinic acetylcholine receptors (nAChRs) mediate fast cholinergic synaptic transmission in the insect nervous system and are targets of widely selling insecticides. We have identified the nAChR gene family from the genome of the malaria mosquito vector, Anopheles gambiae, to be the second complete insect nAChR gene family described following that of Drosophila melanogaster. Like Drosophila, Anopheles possesses 10 nAChR subunits with orthologous relationships evident between the two insects. Interestingly, the Anopheles orthologues of Dbeta2 and Dbeta3 possess the vicinal cysteines that define alpha subunits. As with Dalpha4 and Dalpha6, the Anopheles orthologues are alternatively spliced at equivalent exons. Reverse transcription-polymerase chain reaction analysis shows that RNA A-to-I editing sites conserved between Dalpha6 of Drosophila and alpha7-2 of the tobacco budworm, Heliothis virescens, are not shared with the equivalent nAChR subunit of Anopheles. Indeed, RNA-editing sites identified in functionally significant regions of Dbeta1, Dalpha5, and Dalpha6 are not conserved in the mosquito orthologues, indicating considerable divergence of RNA molecules targeted for editing within the insect order Diptera. These findings shed further light on the diversity of nAChR subunits and may present a useful basis for the development of improved malaria control agents by enhancing our understanding of a validated mosquito insecticide target.

  11. In depth annotation of the Anopheles gambiae mosquito midgut transcriptome

    OpenAIRE

    Padrón, Alejandro; Molina-Cruz, Alvaro; Quinones, Mariam; Ribeiro, José MC; Ramphul, Urvashi; Rodrigues, Janneth; Shen, Kui; Haile, Ashley; Ramirez, José Luis; Barillas-Mury, Carolina

    2014-01-01

    Background Genome sequencing of Anopheles gambiae was completed more than ten years ago and has accelerated research on malaria transmission. However, annotation needs to be refined and verified experimentally, as most predicted transcripts have been identified by comparative analysis with genomes from other species. The mosquito midgut—the first organ to interact with Plasmodium parasites—mounts effective antiplasmodial responses that limit parasite survival and disease transmission. High-th...

  12. Comparative susceptibility to Plasmodium falciparum of the molecular forms M and S of Anopheles gambiae and Anopheles arabiensis

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    Boudin Christian

    2011-09-01

    Full Text Available Abstract Background The different taxa belonging to Anopheles gambiae complex display phenotypic differences that may impact their contribution to malaria transmission. More specifically, their susceptibility to infection, resulting from a co-evolution between parasite and vector, might be different. The aim of this study was to compare the susceptibility of M and S molecular forms of Anopheles gambiae and Anopheles arabiensis to infection by Plasmodium falciparum. Methods F3 progenies of Anopheles gambiae s.l. collected in Senegal were infected, using direct membrane feeding, with P. falciparum gametocyte-containing blood sampled on volunteer patients. The presence of oocysts was determined by light microscopy after 7 days, and the presence of sporozoite by ELISA after 14 days. Mosquito species and molecular forms were identified by PCR. Results The oocyst rate was significantly higher in the molecular S form (79.07% than in the M form (57.81%, Fisher's exact test p Anopheles arabiensis (55.38%, Fisher's exact test vs. S group p An. gambiae S form (1.72 ± 0.26 than in the An. gambiae M form (0.64 ± 0.04, p An. arabiensis group (0.58 ± 0.04, vs. S group, p Anopheles arabiensis 50.85%, Fisher's exact test vs. S group p Conclusion Infected in the same experimental conditions, the molecular form S of An. gambiae is more susceptible to infection by P. falciparum than the molecular form M of An. gambiae and An. arabiensis.

  13. Frequent blood-feeding and restrictive sugar-feeding behavior enhance the malaria vector potential of Anopheles gambiae s.l. and An. funestus (Diptera:Culicidae) in western Kenya.

    Science.gov (United States)

    Beier, J C

    1996-07-01

    Natural blood-feeding and sugar-feeding behaviors were investigated for populations of Anopheles gambiae s.l. and An. funestus Giles at 2 sites in western Kenya. During peak levels of malaria parasite transmission, > 85% of 1,569 indoor-resting females contained fresh blood meals. Findings that up to 55.4% of blood-fed resting females and 72.0% of host-seeking females had either stage IV or V oocytes provided strong evidence that females were refeeding before oviposition. Such gonotrophic discordance was common throughout the year for both An. gambiae s.l. and An. funestus. Determinations of gonotrophic cycles for freshly blood-fed mosquitoes collected inside houses indicated that only 60.0% of 1,287 An. gambiae s.l. and 60.0% of 974 An. funestus oviposited eggs after a single blood meal. The timing of oviposition was irregular as indicated by relatively high coefficients of variation for An. gambiae s.l. (44.0%) and An. funestus (35.9%). Associated with frequent blood feeding was a surprisingly low rate of sugar feeding; only 6.3% of 1,183 indoor-resting and only 14.4% of 236 host-seeking anophelines were positive for fructose. Natural patterns of frequent blood feeding, year-round gonotrophic discordance, irregular oviposition cycles, and limited sugar feeding illustrate that anopheline mosquitoes have complex behavioral and physiologic means for adapting to their environment. In western Kenya, for example, adaptations for frequent blood feeding by An. gambiae s.l. and An. funestus potentiates their ability to transmit malaria parasites, well beyond that predicted by standard measures of vectorial capacity. PMID:8699456

  14. Immunity-related genes and gene families in Anopheles gambiae.

    Science.gov (United States)

    Christophides, George K; Zdobnov, Evgeny; Barillas-Mury, Carolina; Birney, Ewan; Blandin, Stephanie; Blass, Claudia; Brey, Paul T; Collins, Frank H; Danielli, Alberto; Dimopoulos, George; Hetru, Charles; Hoa, Ngo T; Hoffmann, Jules A; Kanzok, Stefan M; Letunic, Ivica; Levashina, Elena A; Loukeris, Thanasis G; Lycett, Gareth; Meister, Stephan; Michel, Kristin; Moita, Luis F; Müller, Hans-Michael; Osta, Mike A; Paskewitz, Susan M; Reichhart, Jean-Marc; Rzhetsky, Andrey; Troxler, Laurent; Vernick, Kenneth D; Vlachou, Dina; Volz, Jennifer; von Mering, Christian; Xu, Jiannong; Zheng, Liangbiao; Bork, Peer; Kafatos, Fotis C

    2002-10-01

    We have identified 242 Anopheles gambiae genes from 18 gene families implicated in innate immunity and have detected marked diversification relative to Drosophila melanogaster. Immune-related gene families involved in recognition, signal modulation, and effector systems show a marked deficit of orthologs and excessive gene expansions, possibly reflecting selection pressures from different pathogens encountered in these insects' very different life-styles. In contrast, the multifunctional Toll signal transduction pathway is substantially conserved, presumably because of counterselection for developmental stability. Representative expression profiles confirm that sequence diversification is accompanied by specific responses to different immune challenges. Alternative RNA splicing may also contribute to expansion of the immune repertoire. PMID:12364793

  15. Mosaic genome architecture of the Anopheles gambiae species complex.

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    Rui Wang-Sattler

    Full Text Available BACKGROUND: Attempts over the last three decades to reconstruct the phylogenetic history of the Anopheles gambiae species complex have been important for developing better strategies to control malaria transmission. METHODOLOGY: We used fingerprint genotyping data from 414 field-collected female mosquitoes at 42 microsatellite loci to infer the evolutionary relationships of four species in the A. gambiae complex, the two major malaria vectors A. gambiae sensu stricto (A. gambiae s.s. and A. arabiensis, as well as two minor vectors, A. merus and A. melas. PRINCIPAL FINDINGS: We identify six taxonomic units, including a clear separation of West and East Africa A. gambiae s.s. S molecular forms. We show that the phylogenetic relationships vary widely between different genomic regions, thus demonstrating the mosaic nature of the genome of these species. The two major malaria vectors are closely related and closer to A. merus than to A. melas at the genome-wide level, which is also true if only autosomes are considered. However, within the Xag inversion region of the X chromosome, the M and two S molecular forms are most similar to A. merus. Near the X centromere, outside the Xag region, the two S forms are highly dissimilar to the other taxa. Furthermore, our data suggest that the centromeric region of chromosome 3 is a strong discriminator between the major and minor malaria vectors. CONCLUSIONS: Although further studies are needed to elucidate the basis of the phylogenetic variation among the different regions of the genome, the preponderance of sympatric admixtures among taxa strongly favor introgression of different genomic regions between species, rather than lineage sorting of ancestral polymorphism, as a possible mechanism.

  16. Population genetic structure of Anopheles arabiensis and Anopheles gambiae in a malaria endemic region of southern Tanzania

    NARCIS (Netherlands)

    K.R. Ng'habi; B.G.J. Knols; Y. Lee; H.M. Ferguson; G.C. Lanzaro

    2011-01-01

    Background: Genetic diversity is a key factor that enables adaptation and persistence of natural populations towards environmental conditions. It is influenced by the interaction of a natural population's dynamics and the environment it inhabits. Anopheles gambiae s.s. and Anopheles arabiensis are t

  17. A proteomic investigation of soluble olfactory proteins in Anopheles gambiae.

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    Guido Mastrobuoni

    Full Text Available Odorant-binding proteins (OBPs and chemosensory proteins (CSPs are small soluble polypeptides that bind semiochemicals in the lymph of insect chemosensilla. In the genome of Anopheles gambiae, 66 genes encode OBPs and 8 encode CSPs. Here we monitored their expression through classical proteomics (2D gel-MS analysis and a shotgun approach. The latter method proved much more sensitive and therefore more suitable for tiny biological samples as mosquitoes antennae and eggs. Females express a larger number and higher quantities of OBPs in their antennae than males (24 vs 19. OBP9 is the most abundant in the antennae of both sexes, as well as in larvae, pupae and eggs. Of the 8 CSPs, 4 were detected in antennae, while SAP3 was the only one expressed in larvae. Our proteomic results are in fairly good agreement with data of RNA expression reported in the literature, except for OBP4 and OBP5, that we could not identify in our analysis, nor could we detect in Western Blot experiments. The relatively limited number of soluble olfactory proteins expressed at relatively high levels in mosquitoes makes further studies on the coding of chemical messages at the OBP level more accessible, providing for few specific targets. Identification of such proteins in Anopheles gambiae might facilitate future studies on host finding behavior in this important disease vector.

  18. Factors affecting fungus-induced larval mortality in Anopheles gambiae and Anopheles stephensi

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    Takken Willem

    2010-01-01

    Full Text Available Abstract Background Entomopathogenic fungi have shown great potential for the control of adult malaria vectors. However, their ability to control aquatic stages of anopheline vectors remains largely unexplored. Therefore, how larval characteristics (Anopheles species, age and larval density, fungus (species and concentration and environmental effects (exposure duration and food availability influence larval mortality caused by fungus, was studied. Methods Laboratory bioassays were performed on the larval stages of Anopheles gambiae and Anopheles stephensi with spores of two fungus species, Metarhizium anisopliae and Beauveria bassiana. For various larval and fungal characteristics and environmental effects the time to death was determined and survival curves established. These curves were compared by Kaplan Meier and Cox regression analyses. Results Beauveria bassiana and Metarhizium anisopliae caused high mortality of An. gambiae and An. stephensi larvae. However, Beauveria bassiana was less effective (Hazard ratio (HR Metarhizium anisopliae. Anopheles stephensi and An. gambiae were equally susceptible to each fungus. Older larvae were less likely to die than young larvae (HR Conclusions This study shows that both fungus species have potential to kill mosquitoes in the larval stage, and that mortality rate depends on fungus species itself, larval stage targeted, larval density and amount of nutrients available to the larvae. Increasing the concentration of fungal spores or reducing the exposure time to spores did not show a proportional increase and decrease in mortality rate, respectively, because the spores clumped together. As a result spores did not provide uniform coverage over space and time. It is, therefore, necessary to develop a formulation that allows the spores to spread over the water surface. Apart from formulation appropriate delivery methods are also necessary to avoid exposing non-target organisms to fungus.

  19. Short report : Development of a molecular assay to detect predation on Anopheles gambiae complex larval stages

    OpenAIRE

    Schielke, E.; Costantini, Carlo; Carchini, G.; Sagnon, N.; J. Powell; Caccone, A

    2007-01-01

    We developed a molecular assay to detect predation on Anopheles gambiae sensu lato (s.l.) mosquitoes. This intergenic spacer ribosomal DNA polymerase chain reaction assay and restriction enzyme analysis uses An. gambiae-specific primers to detect mosquito DNA in the DNA extracts from whole invertebrate predators, which enables identification of species (An. gambiae s.s. versus An. arabiensis) and molecular forms (M versus S in An. gambiae s.s.). We show that An. gambiae s.l. DNA can be detect...

  20. Entomological indices of Anopheles gambiae sensu lato at a rural community in south-west Nigeria

    OpenAIRE

    M.A.E. Noutcha; C.I. Anumdu

    2009-01-01

    Background & objectives: Investigations were conducted to obtain key entomological indices of Anopheles gambiae s.l. at Igbo-Ora, a rural community in south-west Nigeria. Methods: Mosquitoes were caught daily for a week from rooms where tenants had slept the previous night in each of the four months June, July (2001), and August, September (2002). Anopheles gambiae s.l. sibling species were PCR-identified, the blood meal origin was determined by direct ELISA, and the circumsporozoite antigen ...

  1. Improvement of a synthetic lure for Anopheles gambiae using compounds produced by human skin microbiota

    NARCIS (Netherlands)

    Verhulst, N.O.; Mbadi, P.A.; Bukovinszkine-Kiss, G.; Mukabana, W.R.; Loon, van J.J.A.; Takken, W.; Smallegange, R.C.

    2011-01-01

    Background - Anopheles gambiae sensu stricto is considered to be highly anthropophilic and volatiles of human origin provide essential cues during its host-seeking behaviour. A synthetic blend of three human-derived volatiles, ammonia, lactic acid and tetradecanoic acid, attracts A. gambiae. In addi

  2. Behavioural response of the malaria vector Anopheles gambiae to host plant volatiles and synthetic blends

    Science.gov (United States)

    Sugar feeding is critical for survival of malaria vectors and, although discriminative plant feeding previously has been shown to occur in Anopheles gambiae s.s., little is known about the cues mediating attraction to these plants. In this study, we investigated the role of olfaction in An. gambiae ...

  3. Inhibition of Anopheles gambiae odorant receptor function by mosquito repellents.

    Science.gov (United States)

    Tsitoura, Panagiota; Koussis, Konstantinos; Iatrou, Kostas

    2015-03-20

    The identification of molecular targets of insect repellents has been a challenging task, with their effects on odorant receptors (ORs) remaining a debatable issue. Here, we describe a study on the effects of selected mosquito repellents, including the widely used repellent N,N-diethyl-meta-toluamide (DEET), on the function of specific ORs of the African malaria vector Anopheles gambiae. This study, which has been based on quantitative measurements of a Ca(2+)-activated photoprotein biosensor of recombinant OR function in an insect cell-based expression platform and a sequential compound addition protocol, revealed that heteromeric OR (ORx/Orco) function was susceptible to strong inhibition by all tested mosquito repellents except DEET. Moreover, our results demonstrated that the observed inhibition was due to efficient blocking of Orco (olfactory receptor coreceptor) function. This mechanism of repellent action, which is reported for the first time, is distinct from the mode of action of other characterized insect repellents including DEET. PMID:25657000

  4. Organization of olfactory centres in the malaria mosquito Anopheles gambiae

    Science.gov (United States)

    Riabinina, Olena; Task, Darya; Marr, Elizabeth; Lin, Chun-Chieh; Alford, Robert; O'Brochta, David A.; Potter, Christopher J.

    2016-01-01

    Mosquitoes are vectors for multiple infectious human diseases and use a variety of sensory cues (olfactory, temperature, humidity and visual) to locate a human host. A comprehensive understanding of the circuitry underlying sensory signalling in the mosquito brain is lacking. Here we used the Q-system of binary gene expression to develop transgenic lines of Anopheles gambiae in which olfactory receptor neurons expressing the odorant receptor co-receptor (Orco) gene are labelled with GFP. These neurons project from the antennae and maxillary palps to the antennal lobe (AL) and from the labella on the proboscis to the suboesophageal zone (SEZ), suggesting integration of olfactory and gustatory signals occurs in this brain region. We present detailed anatomical maps of olfactory innervations in the AL and the SEZ, identifying glomeruli that may respond to human body odours or carbon dioxide. Our results pave the way for anatomical and functional neurogenetic studies of sensory processing in mosquitoes. PMID:27694947

  5. The role of hemocytes in Anopheles gambiae antiplasmodial immunity.

    Science.gov (United States)

    Ramirez, Jose Luis; Garver, Lindsey S; Brayner, Fábio André; Alves, Luiz Carlos; Rodrigues, Janneth; Molina-Cruz, Alvaro; Barillas-Mury, Carolina

    2014-01-01

    Hemocytes synthesize key components of the mosquito complement-like system, but their role in the activation of antiplasmodial responses has not been established. The effect of activating Toll signaling in hemocytes on Plasmodium survival was investigated by transferring hemocytes or cell-free hemolymph from donor mosquitoes in which the suppressor cactus was silenced. These transfers greatly enhanced antiplasmodial immunity, indicating that hemocytes are active players in the activation of the complement-like system, through an effector/effectors regulated by the Toll pathway. A comparative analysis of hemocyte populations between susceptible G3 and the refractory L3-5 Anopheles gambiae mosquito strains did not reveal significant differences under basal conditions or in response to Plasmodium berghei infection. The response of susceptible mosquitoes to different Plasmodium species revealed similar kinetics following infection with P. berghei,P. yoelii or P. falciparum, but the strength of the priming response was stronger in less compatible mosquito-parasite pairs. The Toll, Imd,STAT or JNK signaling cascades were not essential for the production of the hemocyte differentiation factor (HDF) in response to P. berghei infection, but disruption of Toll, STAT or JNK abolished hemocyte differentiation in response to HDF. We conclude that hemocytes are key mediators of A. gambiae antiplasmodial responses. PMID:23886925

  6. The role of reactive oxygen species on Plasmodium melanotic encapsulation in Anopheles gambiae

    OpenAIRE

    Kumar, Sanjeev; Christophides, George K.; Cantera, Rafael; Charles, Bradley; Han, Yeon Soo; Meister, Stephan; Dimopoulos, George; Kafatos, Fotis C.; Barillas-Mury, Carolina

    2003-01-01

    Malaria transmission depends on the competence of some Anopheles mosquitoes to sustain Plasmodium development (susceptibility). A genetically selected refractory strain of Anopheles gambiae blocks Plasmodium development, melanizing, and encapsulating the parasite in a reaction that begins with tyrosine oxidation, and involves three quantitative trait loci. Morphological and microarray mRNA expression analysis suggest that the refractory and susceptible strains have broad physiological differe...

  7. Bioinformatics-Based Identification of Chemosensory Proteins in African Malaria Mosquito, Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    Zhengxi Li; Zuorui Shen; Jingjiang Zhou; Lin Field

    2003-01-01

    Chemosensory proteins (CSPs) are identifiable by four spatially conserved Cysteine residues in their primary structure or by two disulfide bridges in their tertiary structure according to the previously identified olfactory specific-D related proteins. A genomics- and bioinformatics-based approach is taken in the present study to identify the putative CSPs in the malaria-carrying mosquito, Anopheles gambiae. The results show that five out of the nine annotated candidates are the most possible Anopheles CSPs of A. gambiae. This study lays the foundation for further functional identification of Anopheles CSPs, though all of these candidates need additional experimental verification.

  8. Chloroquine mediated modulation of Anopheles gambiae gene expression.

    Directory of Open Access Journals (Sweden)

    Patrícia Abrantes

    Full Text Available BACKGROUND: Plasmodium development in the mosquito is crucial for malaria transmission and depends on the parasite's interaction with a variety of cell types and specific mosquito factors that have both positive and negative effects on infection. Whereas the defensive response of the mosquito contributes to a decrease in parasite numbers during these stages, some components of the blood meal are known to favor infection, potentiating the risk of increased transmission. The presence of the antimalarial drug chloroquine in the mosquito's blood meal has been associated with an increase in Plasmodium infectivity for the mosquito, which is possibly caused by chloroquine interfering with the capacity of the mosquito to defend against the infection. METHODOLOGY/PRINCIPAL FINDINGS: In this study, we report a detailed survey of the Anopheles gambiae genes that are differentially regulated by the presence of chloroquine in the blood meal, using an A. gambiae cDNA microarray. The effect of chloroquine on transcript abundance was evaluated separately for non-infected and Plasmodium berghei-infected mosquitoes. Chloroquine was found to affect the abundance of transcripts that encode proteins involved in a variety of processes, including immunity, apoptosis, cytoskeleton and the response to oxidative stress. This pattern of differential gene expression may explain the weakened mosquito defense response which accounts for the increased infectivity observed in chloroquine-treated mosquitoes. CONCLUSIONS/SIGNIFICANCE: The results of the present study suggest that chloroquine can interfere with several putative mosquito mechanisms of defense against Plasmodium at the level of gene expression and highlight the need for a better understanding of the impacts of antimalarial agents on parasite transmission.

  9. Anopheles gambiae Purine Nucleoside Phosphorylase: Catalysis, Structure, and Inhibition

    Energy Technology Data Exchange (ETDEWEB)

    Taylor,E.; Rinaldo-Matthis, A.; Li, L.; Ghanem, M.; Hazleton, K.; Cassera, M.; Almo, S.; Schramm, V.

    2007-01-01

    The purine salvage pathway of Anopheles gambiae, a mosquito that transmits malaria, has been identified in genome searches on the basis of sequence homology with characterized enzymes. Purine nucleoside phosphorylase (PNP) is a target for the development of therapeutic agents in humans and purine auxotrophs, including malarial parasites. The PNP from Anopheles gambiae (AgPNP) was expressed in Escherichia coli and compared to the PNPs from Homo sapiens (HsPNP) and Plasmodium falciparum (PfPNP). AgPNP has kcat values of 54 and 41 s-1 for 2'-deoxyinosine and inosine, its preferred substrates, and 1.0 s-1 for guanosine. However, the chemical step is fast for AgPNP at 226 s-1 for guanosine in pre-steady-state studies. 5'-Deaza-1'-aza-2'-deoxy-1'-(9-methylene)-Immucillin-H (DADMe-ImmH) is a transition-state mimic for a 2'-deoxyinosine ribocation with a fully dissociated N-ribosidic bond and is a slow-onset, tight-binding inhibitor with a dissociation constant of 3.5 pM. This is the tightest-binding inhibitor known for any PNP, with a remarkable Km/Ki* of 5.4 x 107, and is consistent with enzymatic transition state predictions of enhanced transition-state analogue binding in enzymes with enhanced catalytic efficiency. Deoxyguanosine is a weaker substrate than deoxyinosine, and DADMe-Immucillin-G is less tightly bound than DADMe-ImmH, with a dissociation constant of 23 pM for AgPNP as compared to 7 pM for HsPNP. The crystal structure of AgPNP was determined in complex with DADMe-ImmH and phosphate to a resolution of 2.2 Angstroms to reveal the differences in substrate and inhibitor specificity. The distance from the N1' cation to the phosphate O4 anion is shorter in the AgPNP{center_dot}DADMe-ImmH{center_dot}PO4 complex than in HsPNP{center_dot}DADMe-ImmH{center_dot}SO4, offering one explanation for the stronger inhibitory effect of DADMe-ImmH for AgPNP.

  10. Comparative genome and proteome analysis of Anopheles gambiae and Drosophila melanogaster.

    Science.gov (United States)

    Zdobnov, Evgeny M; von Mering, Christian; Letunic, Ivica; Torrents, David; Suyama, Mikita; Copley, Richard R; Christophides, George K; Thomasova, Dana; Holt, Robert A; Subramanian, G Mani; Mueller, Hans-Michael; Dimopoulos, George; Law, John H; Wells, Michael A; Birney, Ewan; Charlab, Rosane; Halpern, Aaron L; Kokoza, Elena; Kraft, Cheryl L; Lai, Zhongwu; Lewis, Suzanna; Louis, Christos; Barillas-Mury, Carolina; Nusskern, Deborah; Rubin, Gerald M; Salzberg, Steven L; Sutton, Granger G; Topalis, Pantelis; Wides, Ron; Wincker, Patrick; Yandell, Mark; Collins, Frank H; Ribeiro, Jose; Gelbart, William M; Kafatos, Fotis C; Bork, Peer

    2002-10-01

    Comparison of the genomes and proteomes of the two diptera Anopheles gambiae and Drosophila melanogaster, which diverged about 250 million years ago, reveals considerable similarities. However, numerous differences are also observed; some of these must reflect the selection and subsequent adaptation associated with different ecologies and life strategies. Almost half of the genes in both genomes are interpreted as orthologs and show an average sequence identity of about 56%, which is slightly lower than that observed between the orthologs of the pufferfish and human (diverged about 450 million years ago). This indicates that these two insects diverged considerably faster than vertebrates. Aligned sequences reveal that orthologous genes have retained only half of their intron/exon structure, indicating that intron gains or losses have occurred at a rate of about one per gene per 125 million years. Chromosomal arms exhibit significant remnants of homology between the two species, although only 34% of the genes colocalize in small "microsyntenic" clusters, and major interarm transfers as well as intra-arm shuffling of gene order are detected. PMID:12364792

  11. Swarming and mating activity of Anopheles gambiae mosquitoes in semi-field enclosures.

    Science.gov (United States)

    Achinko, D; Thailayil, J; Paton, D; Mireji, P O; Talesa, V; Masiga, D; Catteruccia, F

    2016-03-01

    Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is the major Afro-tropical vector of malaria. Novel strategies proposed for the elimination and eradication of this mosquito vector are based on the use of genetic approaches, such as the sterile insect technique (SIT). These approaches rely on the ability of released males to mate with wild females, and depend on the application of effective protocols to assess the swarming and mating behaviours of laboratory-reared insects prior to their release. The present study evaluated whether large semi-field enclosures can be utilized to study the ability of males from a laboratory colony to respond to natural environmental stimuli and initiate normal mating behaviour. Laboratory-reared males exhibited spatiotemporally consistent swarming behaviour within the study enclosures. Swarm initiation, peak and termination time closely tracked sunset. Comparable insemination rates were observed in females captured in copula in the semi-field cages relative to females in small laboratory cages. Oviposition rates after blood feeding were also similar to those observed in laboratory settings. The data suggest that outdoor enclosures are suitable for studying swarming and mating in laboratory-bred males in field-like settings, providing an important reference for future studies aimed at assessing the comparative mating ability of strains for SIT and other vector control strategies. PMID:26508420

  12. Inactivation of Anopheles gambiae Glutathione Transferase ε2 by Epiphyllocoumarin

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    Patience Marimo

    2016-01-01

    Full Text Available Glutathione transferases (GSTs are part of a major family of detoxifying enzymes that can catalyze the reductive dehydrochlorination of dichlorodiphenyltrichloroethane (DDT. The delta and epsilon classes of insect GSTs have been implicated in conferring resistance to this insecticide. In this study, the inactivation of Anopheles gambiae GSTε2 by epiphyllocoumarin (Tral 1 was investigated. Recombinant AgGSTε2 was expressed in Escherichia coli cells containing a pET3a-AGSTε2 plasmid and purified by affinity chromatography. Tral 1 was shown to inactivate GSTε2 both in a time-dependent manner and in a concentration-dependent manner. The half-life of GSTε2 in the presence of 25 μM ethacrynic acid (ETA was 22 minutes and with Tral 1 was 30 minutes, indicating that Tral 1 was not as efficient as ETA as an inactivator. The inactivation parameters kinact and KI were found to be 0.020 ± 0.001 min−1 and 7.5 ± 2.1 μM, respectively, after 90 minutes of incubation. Inactivation of GSTε2 by Tral 1 implies that Tral 1 covalently binds to this enzyme in vitro and would be expected to exhibit time-dependent effects on the enzyme in vivo. Tral 1, therefore, would produce irreversible effects when used together with dichlorodiphenyltrichloroethane (DDT in malaria control programmes where resistance is mediated by GSTs.

  13. Innate immunity against malaria parasites in Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    Yang Chenand; Zhi-Hui Weng; Liangbiao Zheng

    2008-01-01

    Malaria continues to exert a huge toll in the world today, causing approximately 400 million cases and killing between 1-2 million people annually. Most of the malaria burden is borne by countries in Africa. For this reason, the major vector for malaria in this continent, Anopheles gambiae, is under intense study. With the completion of the draft sequence of this important vector, efforts are underway to develop novel control strategies.One promising area is to harness the power of the innate immunity of this mosquito species to block the transmission of the malaria parasites. Recent studies have demonstrated that Toll and Imd signaling pathways and other immunity-related genes (encoding proteins possibly function in recognition or as effector molecules) play significant roles in two different arms of innate immunity: level of infection intensity and melanization of Plasmodium oocysts.The challenges in the future are to understand how the functions of these different genes are coordinated in defense against malaria parasites, and if different arms of innate immunity are cross-regulated or coordinated.

  14. Larvicidal Effects of a Neem (Azadirachta indica) Oil Formulation on the Malaria Vector Anopheles Gambiae.

    OpenAIRE

    Knols Bart GJ; Okumu Fredros O; Fillinger Ulrike

    2007-01-01

    Abstract Background Larviciding is a key strategy used in many vector control programmes around the world. Costs could be reduced if larvicides could be manufactured locally. The potential of natural products as larvicides against the main African malaria vector, Anopheles gambiae s.s was evaluated. Methods To assess the larvicidal efficacy of a neem (Azadirachta indica) oil formulation (azadirachtin content of 0.03% w/v) on An. gambiae s.s., larvae were exposed as third and fourth instars to...

  15. The Population Genomics of Trans-Specific Inversion Polymorphisms in Anopheles gambiae

    OpenAIRE

    White, Bradley J.; Cheng, Changde; Sangaré, Djibril; Lobo, Neil F.; Collins, Frank H.; Besansky, Nora J

    2009-01-01

    In the malaria mosquito Anopheles gambiae polymorphic chromosomal inversions may play an important role in adaptation to environmental variation. Recently, we used microarray-based divergence mapping combined with targeted resequencing to map nucleotide differentiation between alternative arrangements of the 2La inversion. Here, we applied the same technique to four different polymorphic inversions on the 2R chromosome of An. gambiae. Surprisingly, divergence was much lower between alternativ...

  16. Evidence of natural Wolbachia infections in field populations of Anopheles gambiae

    Science.gov (United States)

    Baldini, Francesco; Segata, Nicola; Pompon, Julien; Marcenac, Perrine; Robert Shaw, W.; Dabiré, Roch K.; Diabaté, Abdoulaye; Levashina, Elena A.; Catteruccia, Flaminia

    2014-01-01

    Wolbachia are maternally transmitted intracellular bacteria that invade insect populations by manipulating their reproduction and immunity and thus limiting the spread of numerous human pathogens. Experimental Wolbachia infections can reduce Plasmodium numbers in Anopheles mosquitoes in the laboratory, however, natural Wolbachia infections in field anophelines have never been reported. Here we show evidence of Wolbachia infections in Anopheles gambiae in Burkina Faso, West Africa. Sequencing of the 16S rRNA gene identified Wolbachia sequences in both female and male germlines across two seasons, and determined that these sequences are vertically transmitted from mother to offspring. Whole-genome sequencing of positive samples suggests that the genetic material identified in An. gambiae belongs to a novel Wolbachia strain, related to but distinct from strains infecting other arthropods. The evidence of Wolbachia infections in natural Anopheles populations promotes further investigations on the possible use of natural Wolbachia–Anopheles associations to limit malaria transmission. PMID:24905191

  17. Immune factor Gambif1, a new rel family member from the human malaria vector, Anopheles gambiae.

    OpenAIRE

    Barillas-Mury, C; Charlesworth, A.; Gross, I; Richman, A; Hoffmann, J A; Kafatos, F C

    1996-01-01

    A novel rel family member, Gambif1 (gambiae immune factor 1), has been cloned from the human malaria vector, Anopheles gambiae, and shown to be most similar to Drosophila Dorsal and Dif. Gambif1 protein is translocated to the nucleus in fat body cells in response to bacterial challenge, although the mRNA is present at low levels at all developmental stages and is not induced by infection. DNA binding activity to the kappaB-like sites in the A.gambiae Defensin and the Drosophila Diptericin and...

  18. Development of a molecular assay to detect predation on Anopheles gambiae complex larval stages.

    Science.gov (United States)

    Schielke, Erika; Costantini, Carlo; Carchini, Gianmaria; Sagnon, N'falé; Powell, Jeffrey; Caccone, Adalgisa

    2007-09-01

    We developed a molecular assay to detect predation on Anopheles gambiae sensu lato (s.l.) mosquitoes. This intergenic spacer ribosomal DNA polymerase chain reaction assay and restriction enzyme analysis uses An. gambiae-specific primers to detect mosquito DNA in the DNA extracts from whole invertebrate predators, which enables identification of species (An. gambiae s.s. versus An. arabiensis) and molecular forms (M versus S in An. gambiae s.s.). We show that An. gambiae s.l. DNA can be detected after ingestion by members of the families Lestidae (order Odonata) after four hours, Libellulidae (order Odonata) after six hours, and Notonectidae (order Hemiptera) after 24 hours. This method is an improvement over previously published methods because of ease of execution and increased time of detection after ingestion. PMID:17827361

  19. A High-Affinity Adenosine Kinase from Anopheles Gambiae

    Energy Technology Data Exchange (ETDEWEB)

    M Cassera; M Ho; E Merino; E Burgos; A Rinaldo-Matthis; S Almo; V Schramm

    2011-12-31

    Genome analysis revealed a mosquito orthologue of adenosine kinase in Anopheles gambiae (AgAK; the most important vector for the transmission of Plasmodium falciparum in Africa). P. falciparum are purine auxotrophs and do not express an adenosine kinase but rely on their hosts for purines. AgAK was kinetically characterized and found to have the highest affinity for adenosine (K{sub m} = 8.1 nM) of any known adenosine kinase. AgAK is specific for adenosine at the nucleoside site, but several nucleotide triphosphate phosphoryl donors are tolerated. The AgAK crystal structure with a bound bisubstrate analogue Ap{sub 4}A (2.0 {angstrom} resolution) reveals interactions for adenosine and ATP and the geometry for phosphoryl transfer. The polyphosphate charge is partly neutralized by a bound Mg{sup 2+} ion and an ion pair to a catalytic site Arg. The AgAK structure consists of a large catalytic core in a three-layer {alpha}/{beta}/{alpha} sandwich, and a small cap domain in contact with adenosine. The specificity and tight binding for adenosine arise from hydrogen bond interactions of Asn14, Leu16, Leu40, Leu133, Leu168, Phe168, and Thr171 and the backbone of Ile39 and Phe168 with the adenine ring as well as through hydrogen bond interactions between Asp18, Gly64, and Asn68 and the ribosyl 2'- and 3'-hydroxyl groups. The structure is more similar to that of human adenosine kinase (48% identical) than to that of AK from Toxoplasma gondii (31% identical). With this extraordinary affinity for AgAK, adenosine is efficiently captured and converted to AMP at near the diffusion limit, suggesting an important role for this enzyme in the maintenance of the adenine nucleotide pool. mRNA analysis verifies that AgAK transcripts are produced in the adult insects.

  20. Unexpected high losses of Anopheles gambiae larvae due to rainfall.

    Directory of Open Access Journals (Sweden)

    Krijn P Paaijmans

    Full Text Available BACKGROUND: Immature stages of the malaria mosquito Anopheles gambiae experience high mortality, but its cause is poorly understood. Here we study the impact of rainfall, one of the abiotic factors to which the immatures are frequently exposed, on their mortality. METHODOLOGY/PRINCIPAL FINDINGS: We show that rainfall significantly affected larval mosquitoes by flushing them out of their aquatic habitat and killing them. Outdoor experiments under natural conditions in Kenya revealed that the additional nightly loss of larvae caused by rainfall was on average 17.5% for the youngest (L1 larvae and 4.8% for the oldest (L4 larvae; an additional 10.5% (increase from 0.9 to 11.4% of the L1 larvae and 3.3% (from 0.1 to 3.4% of the L4 larvae were flushed away and larval mortality increased by 6.9% (from 4.6 to 11.5% and 1.5% (from 4.1 to 5.6% for L1 and L4 larvae, respectively, compared to nights without rain. On rainy nights, 1.3% and 0.7% of L1 and L4 larvae, respectively, were lost due to ejection from the breeding site. CONCLUSIONS/SIGNIFICANCE: This study demonstrates that immature populations of malaria mosquitoes suffer high losses during rainfall events. As these populations are likely to experience several rain showers during their lifespan, rainfall will have a profound effect on the productivity of mosquito breeding sites and, as a result, on the transmission of malaria. These findings are discussed in the light of malaria risk and changing rainfall patterns in response to climate change.

  1. Larvicidal effects of a neem (Azadirachta indica) oil formulation on the malaria vector Anopheles gambiae

    NARCIS (Netherlands)

    Okumu, F.O.; Knols, B.G.J.; Fillinger, U.

    2007-01-01

    Background - Larviciding is a key strategy used in many vector control programmes around the world. Costs could be reduced if larvicides could be manufactured locally. The potential of natural products as larvicides against the main African malaria vector, Anopheles gambiae s.s was evaluated. Method

  2. Control of pyrethroid-resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes with chlorfenapyr in Benin

    NARCIS (Netherlands)

    N'Guessan, R.; Boko, P.; Odjo, A.; Knols, B.G.J.; Akogbeto, M.; Rowland, M.

    2009-01-01

    Objective To compare the efficacy of chlorfenapyr applied on mosquito nets and as an indoor residual spray against populations of Anopheles gambiae and Culex quinquefasciatus in an area of Benin that shows problematic levels of pyrethroid resistance. Method Eight-week trial conducted in experimental

  3. Olfaction in the malaria mosquito Anopheles gambiae : Electrophysiology and identification of kairomones

    NARCIS (Netherlands)

    Meijerink, J.

    1999-01-01

    Female mosquitoes of the species Anopheles gambiae Giles sensu stricto are important vectors of human malaria in Africa. It is generally assumed that they locate their human host by odours. These odours are detected by olfactory receptor neurons situated within cuticular extensions on the antenna. T

  4. Inhibition of host-seeking response and olfactory responsiveness in Anopheles gambiae following blood feeding

    NARCIS (Netherlands)

    Takken, W.; Loon, van J.J.A.; Adam, W.

    2001-01-01

    The effect of a single blood meal on the host-seeking response of Anopheles gambiae was investigated in the laboratory using a behavioural bioassay, whereas possible changes at the chemosensory level were monitored using electroantennogram recording (EAG). To avoid the possible confounding effect of

  5. Species-specific chemosensory gene expression in the olfactory organs of the malaria vector Anopheles gambiae

    NARCIS (Netherlands)

    Hodges, Theresa K.; Cosme, Luciano V.; Athrey, Giridhar; Pathikonda, Sharmila; Takken, Willem; Slotman, Michel A.

    2014-01-01

    Background: The malaria mosquito Anopheles gambiae has a high preference for human hosts, a characteristic that contributes greatly to its capacity for transmitting human malaria. A sibling species, An. quadriannulatus, has a quite different host preference and feeds mostly on bovids. For this re

  6. Identification of field caught Anopheles gambiae s.s. and Anopheles arabiensis by TaqMan single nucleotide polymorphism genotyping

    Directory of Open Access Journals (Sweden)

    Bayoh Nabie M

    2007-02-01

    Full Text Available Abstract Background Identification of Anopheles gambiae s.s. and Anopheles arabiensis from field-collected Anopheles gambiae s.l. is often necessary in basic and applied research, and in operational control programmes. The currently accepted method involves use of standard polymerase chain reaction amplification of ribosomal DNA (rDNA from the 3' 28S to 5' intergenic spacer region of the genome, and visual confirmation of amplicons of predicted size on agarose gels, after electrophoresis. This report describes development and evaluation of an automated, quantitative PCR method based upon TaqMan™ single nucleotide polymorphism (SNP genotyping. Methods Standard PCR, and TaqMan SNP genotyping with newly designed primers and fluorophore-labeled probes hybridizing to sequences of complementary rDNA specific for either An. gambiae s.s. or An. arabiensis, were conducted in three experiments involving field-collected An. gambiae s.l. from western Kenya, and defined laboratory strains. DNA extraction was from a single leg, sonicated for five minutes in buffer in wells of 96-well PCR plates. Results TaqMan SNP genotyping showed a reaction success rate, sensitivity, and species specificity comparable to that of standard PCR. In an extensive field study, only 29 of 3,041 (0.95% were determined to be hybrids by TaqMan (i.e., having rDNA sequences from both species, however, all but one were An. arabiensis by standard PCR, suggesting an acceptably low (ca. 1% error rate for TaqMan genotyping in mistakenly identifying species hybrids. Conclusion TaqMan SNP genotyping proved to be a sensitive and rapid method for identification of An. gambiae s.l. and An. arabiensis, with a high success rate, specific results, and congruence with the standard PCR method.

  7. African water storage pots for the delivery of the Entomopathogenic fungus Metarhizium anisopliae to the Malaria vectors Anopheles gambiae s.s. and Anopheles funestus

    NARCIS (Netherlands)

    Farenhorst, M.; Farina, D.; Scholte, E.J.; Takken, W.; Hunt, R.H.; Coetzee, M.; Knols, B.G.J.

    2008-01-01

    We studied the use of African water storage pots for point source application of Metarhizium anisopliae against the malaria vectors Anopheles gambiae s.s. and An. funestus. Clay pots were shown to be attractive resting sites for male and female An. gambiae s.s. and were not repellent after impregnat

  8. Microsporidium Infecting Anopheles supepictus (Diptera: Culicidae Larvae

    Directory of Open Access Journals (Sweden)

    Seyed-Mohammad Omrani

    2016-01-01

    Full Text Available Background: Microsporidia are known to infect a wide variety of animals including mosquitoes (Diptera: Cu­licidae. In a recent study on the mosquito fauna of Chahar Mahal and Bakhtiari Province, at the central western part of Iran, a few larvae of Anopheles superpictus were infected with a microsporidium-resembled microorganism. Cur­rent investigation deals with the identification of the responsible microorganism at the genus level.Methods: Fresh infected larvae were collected from the field. After determining the species identity they were dis­sected to extract their infective contents. Wet preparations were checked for general appearance and the size of the pathogenic microorganism. Fixed preparations were stained with Geimsa and Ryan-Blue modified Trichrome tech­niques to visualize further morphological characters. The obtained light microscopy data were used in the identifica­tion process.Results: The infected larvae were bulged by a whitish material filling the involved segments corresponding to a microsporidium infection. Bottle-shaped semioval spores ranged 4.33±0.19×2.67±0.12 and 4.18±0.43×2.45±0.33 micron in wet and fixed preparations, respectively. They were mostly arranged in globular structures comprised of 8 spores. These data was in favor of a species from the genus Parathelohania in the family Ambliosporidae.Conclusion: This is the first report of a microsporidium infection in An. superpictus. The causative agent is diag­nosed as a member of the genus Parathelohania. Further identification down to the species level needs to determine its ultrastructural characteristics and the comparative analysis of ss rRNA sequence data. It is also necessary to un­derstand the detail of the components of the transmission cycle.

  9. Seasonality and Locality Affect the Diversity of Anopheles gambiae and Anopheles coluzzii Midgut Microbiota from Ghana.

    Science.gov (United States)

    Akorli, Jewelna; Gendrin, Mathilde; Pels, Nana Adjoa P; Yeboah-Manu, Dorothy; Christophides, George K; Wilson, Michael D

    2016-01-01

    Symbiotic bacteria can have important implications in the development and competence of disease vectors. In Anopheles mosquitoes, the composition of the midgut microbiota is largely influenced by the larval breeding site, but the exact factors shaping this composition are currently unknown. Here, we examined whether the proximity to urban areas and seasons have an impact on the midgut microbial community of the two major malaria vectors in Africa, An. coluzzii and An. gambiae. Larvae and pupae were collected from selected habitats in two districts of Ghana during the dry and rainy season periods. The midgut microbiota of adults that emerged from these collections was determined by 454-pyrosequencing of the 16S ribosomal DNA. We show that in both mosquito species, Shewanellaceae constituted on average of 54% and 73% of the midgut microbiota from each site in the dry and rainy season, respectively. Enterobacteriaceae was found in comparatively low abundance below 1% in 22/30 samples in the dry season, and in 25/38 samples in the rainy season. Our data indicate that seasonality and locality significantly affect both the diversity of microbiota and the relative abundance of bacterial families with a positive impact of dry season and peri-urban settings.

  10. Seasonality and Locality Affect the Diversity of Anopheles gambiae and Anopheles coluzzii Midgut Microbiota from Ghana

    Science.gov (United States)

    Gendrin, Mathilde; Pels, Nana Adjoa P.; Yeboah-Manu, Dorothy; Christophides, George K.; Wilson, Michael D.

    2016-01-01

    Symbiotic bacteria can have important implications in the development and competence of disease vectors. In Anopheles mosquitoes, the composition of the midgut microbiota is largely influenced by the larval breeding site, but the exact factors shaping this composition are currently unknown. Here, we examined whether the proximity to urban areas and seasons have an impact on the midgut microbial community of the two major malaria vectors in Africa, An. coluzzii and An. gambiae. Larvae and pupae were collected from selected habitats in two districts of Ghana during the dry and rainy season periods. The midgut microbiota of adults that emerged from these collections was determined by 454-pyrosequencing of the 16S ribosomal DNA. We show that in both mosquito species, Shewanellaceae constituted on average of 54% and 73% of the midgut microbiota from each site in the dry and rainy season, respectively. Enterobacteriaceae was found in comparatively low abundance below 1% in 22/30 samples in the dry season, and in 25/38 samples in the rainy season. Our data indicate that seasonality and locality significantly affect both the diversity of microbiota and the relative abundance of bacterial families with a positive impact of dry season and peri-urban settings. PMID:27322614

  11. Identification of four evolutionarily related G protein-coupled receptors from the malaria mosquito Anopheles gambiae

    DEFF Research Database (Denmark)

    Belmont, Martin; Cazzamali, Giuseppe; Williamson, Michael;

    2006-01-01

    The mosquito Anopheles gambiae is an important vector for malaria, which is one of the most serious human parasitic diseases in the world, causing up to 2.7 million deaths yearly. To contribute to our understanding of A. gambiae and to the transmission of malaria, we have now cloned four evolutio......The mosquito Anopheles gambiae is an important vector for malaria, which is one of the most serious human parasitic diseases in the world, causing up to 2.7 million deaths yearly. To contribute to our understanding of A. gambiae and to the transmission of malaria, we have now cloned four...... evolutionarily related G protein-coupled receptors (GPCRs) from this mosquito and expressed them in Chinese hamster ovary cells. After screening of a library of thirty-three insect or other invertebrate neuropeptides and eight biogenic amines, we could identify (de-orphanize) three of these GPCRs as...... relationship to the A. gambiae and other insect AKH receptors suggested that it is a receptor for an AKH-like peptide. This is the first published report on evolutionarily related AKH, corazonin, and CCAP receptors in mosquitoes....

  12. Entomological indices of Anopheles gambiae sensu lato at a rural community in south-west Nigeria

    Directory of Open Access Journals (Sweden)

    M.A.E. Noutcha

    2009-02-01

    Full Text Available Background & objectives: Investigations were conducted to obtain key entomological indices of Anopheles gambiae s.l. at Igbo-Ora, a rural community in south-west Nigeria. Methods: Mosquitoes were caught daily for a week from rooms where tenants had slept the previous night in each of the four months June, July (2001, and August, September (2002. Anopheles gambiae s.l. sibling species were PCR-identified, the blood meal origin was determined by direct ELISA, and the circumsporozoite antigen by sandwich ELISA. Mean weekly rates were calculated. Results: The mean human biting rates were 0.90 and 1.6 in 2001 and 2002 respectively. The mean weekly anthropophilic rates for An. gambiae s.l. were 82 and 86% in 2001 and 2002 respectively; they were high in An. gambiae s.s., An. arabiensis and non-identified species in the complex. The mean weekly circumsporozoite rates were 6.70% in 2001 and 6.30% in 2002. The mean weekly entomological inoculation rates (EIR were 4.95 and 5.05 in 2001 and 2002 respectively; the seasonal (6-month rates were high: 128.7 in 2001 and 131.3 in 2002, compared to data from other rural communities on the continent. Interpretation & conclusion: The implications of these findings on the role of An. gambiae s.l. in the holoendemicity of malaria at Igbo-Ora are discussed.

  13. Identification of one capa and two pyrokinin receptors from the malaria mosquito Anopheles gambiae

    DEFF Research Database (Denmark)

    Olsen, Stine S; Cazzamali, Giuseppe; Williamson, Michael;

    2007-01-01

    We cloned the cDNA of three evolutionarily related G protein-coupled receptors from the malaria mosquito Anopheles gambiae and functionally expressed them in Chinese hamster ovary cells. One receptor, Ang-Capa-R, was only activated by the two Anopheles capa neuropeptides Ang-capa-1 (GPTVGLFAFPRVa......We cloned the cDNA of three evolutionarily related G protein-coupled receptors from the malaria mosquito Anopheles gambiae and functionally expressed them in Chinese hamster ovary cells. One receptor, Ang-Capa-R, was only activated by the two Anopheles capa neuropeptides Ang-capa-1...... (GPTVGLFAFPRVamide) and Ang-capa-2 (pQGLVPFPRVamide) with EC(50) values of 8.6x10(-9)M and 3.3x10(-9)M, respectively, but not by any other known mosquito neuropeptide. The second receptor, Ang-PK-1-R, was selectively activated by the Anopheles pyrokinin-1 peptides Ang-PK-1-1 (AGGTGANSAMWFGPRLamide) and Ang-PK-1......-2 (AAAMWFGPRLamide) with EC(50) values of 3.3x10(-8)M and 2.5x10(-8)M, respectively, but not by mosquito capa or pyrokinin-2 peptides. For the third receptor, Ang-PK-2-R, the most potent ligands were the pyrokinin-2 peptides Ang-PK-2-1 (DSVGENHQRPPFAPRLamide) and Ang-PK-2-2 (NLPFSPRLamide) with EC(50) values of 5.2x...

  14. Molecular identification of a myosuppressin receptor from the malaria mosquito Anopheles gambiae

    DEFF Research Database (Denmark)

    Schöller, Susanne; Belmont, Martin; Cazzamali, Giuseppe;

    2005-01-01

    The insect myosuppressins (X1DVX2HX3FLRFamide) are neuropeptides that generally block insect muscle activities. We have used the genomic sequence information from the malaria mosquito Anopheles gambiae Genome Project to clone a G protein-coupled receptor that was closely related to the two...... previously cloned and characterized myosuppressin receptors from Drosophila [Proc. Natl. Acad. Sci. USA 100 (2003) 9808]. The mosquito receptor cDNA was expressed in Chinese hamster ovary cells and was found to be activated by low concentrations of Anopheles myosuppressin (TDVDHVFLRFamide; EC50, 1.6 x 10...... identification of a mosquito neuropeptide receptor....

  15. Limited usefulness of microsatellite markers from the malaria vector Anopheles gambiae when applied to the closely related species Anopheles melas.

    Science.gov (United States)

    Deitz, Kevin C; Reddy, Vamsi P; Reddy, Michael R; Satyanarayanah, Neha; Lindsey, Michael W; Overgaard, Hans J; Jawara, Musa; Caccone, Adalgisa; Slotman, Michel A

    2012-07-01

    Anopheles melas is a brackish water mosquito found in coastal West Africa where it is a dominant malaria vector locally. In order to facilitate genetic studies of this species, 45 microsatellite loci originally developed for Anopheles gambiae were sequenced in An. melas. Those that were suitable based on repeat number and flanking regions were examined in 2 natural populations from Equatorial Guinea. Only 15 loci were eventually deemed suitable as polymorphic markers in An. melas populations. These loci were screened in 4 populations from a wider geographic range. Heterozygosity estimates ranged from 0.18 to 0.79, and 2.5-15 average alleles were observed per locus, yielding 13 highly polymorphic markers and 2 loci with lower variability. To examine the usefulness of microsatellite markers when applied in a sibling species, the original An. gambiae specific markers were used to amplify 5 loci in An. melas. Null alleles were found for 1 An. gambiae marker. We discuss the pitfalls of using microsatellite loci across closely related species and conclude that in addition to the problem of null alleles associated with this practice, many loci may prove to be of very limited use as polymorphic markers even when used in a sibling species. PMID:22593601

  16. Integrated genetic map of Anopheles gambiae: use of RAPD polymorphisms for genetic, cytogenetic and STS landmarks.

    Science.gov (United States)

    Dimopoulos, G; Zheng, L; Kumar, V; della Torre, A; Kafatos, F C; Louis, C

    1996-06-01

    Randomly amplified polymorphic DNA (RAPD) markers have been integrated in the genetic and cytogenetic maps of the malaria vector mosquito, Anopheles gambiae. Fifteen of these markers were mapped by recombination, relative to microsatellite markers that had been mapped previously. Thirty-four gel-purified RAPD bands were cloned and sequenced, generating sequence tagged sites (STSs) that can be used as entry points to the A. gambiae genome. Thirty one of these STSs were localized on nurse cell polytene chromosomes through their unique hybridization signal in in situ hybridization experiments. Five STSs map close to the breakpoints of polymorphic inversions, which are notable features of the Anopheles genome. The usefulness and limitations of this integrated mosquito map are discussed. PMID:8725241

  17. Olfaction in the malaria mosquito Anopheles gambiae : Electrophysiology and identification of kairomones

    OpenAIRE

    Meijerink, J

    1999-01-01

    Female mosquitoes of the species Anopheles gambiae Giles sensu stricto are important vectors of human malaria in Africa. It is generally assumed that they locate their human host by odours. These odours are detected by olfactory receptor neurons situated within cuticular extensions on the antenna. These cuticular extensions, called sensilla, contain numerous pores through which the odours can enter the sensillum and reach the olfactory receptor neuron membrane. Despite the fact that these mos...

  18. Rhodopsin coexpression in UV photoreceptors of Aedes aegypti and Anopheles gambiae mosquitoes

    OpenAIRE

    Hu, Xiaobang; Leming, Matthew T; Whaley, Michelle A.; O'Tousa, Joseph E.

    2014-01-01

    Differential rhodopsin gene expression within specialized R7 photoreceptor cells divides the retinas of Aedes aegypti and Anopheles gambiae mosquitoes into distinct domains. The two species express the rhodopsin orthologs Aaop8 and Agop8, respectively, in a large subset of these R7 photoreceptors that function as ultraviolet receptors. We show here that a divergent subfamily of mosquito rhodopsins, Aaop10 and Agop10, is coexpressed in these R7 photoreceptors. The properties of the A. aegypti ...

  19. Anopheles gambiae mosquito isolated neurons : a new biological model for optimizing insecticide/repellent efficacy

    OpenAIRE

    Lavialle-Defaix, C.; Apaire-Marchais, V; Legros, C.; Pennetier, Cédric; Mohamed, A; P. Licznar; Corbel, Vincent; Lapied, B

    2011-01-01

    To understand better the mode of action of insecticides and repellents used in vector-borne diseases control, we developed a new biological model based on mosquito neurons isolated from adults Anopheles gambiae heads. This cellular model is well adapted to multidisciplinary approaches: electrophysiology, pharmacology, molecular biology and biochemical assays. Using RT-PCR, we demonstrated that isolated neurons express the nicotinic acetylcholine receptor subunit alpha 1 (Ag alpha 1 nAchR), tw...

  20. Extensive circadian and light regulation of the transcriptome in the malaria mosquito Anopheles gambiae

    OpenAIRE

    Rund, Samuel SC; James E. Gentile; Duffield, Giles E.

    2013-01-01

    Background Mosquitoes exhibit 24 hr rhythms in flight activity, feeding, reproduction and development. To better understand the molecular basis for these rhythms in the nocturnal malaria vector Anopheles gambiae, we have utilized microarray analysis on time-of-day specific collections of mosquitoes over 48 hr to explore the coregulation of gene expression rhythms by the circadian clock and light, and compare these with the 24 hr rhythmic gene expression in the diurnal Aedes aegypti dengue vec...

  1. Ecological Genomics of Anopheles gambiae Along a Latitudinal Cline: A Population-Resequencing Approach

    OpenAIRE

    Cheng, Changde; White, Bradley J.; Kamdem, Colince; Mockaitis, Keithanne; Costantini, Carlo; Matthew W Hahn; Besansky, Nora J

    2012-01-01

    The association between fitness-related phenotypic traits and an environmental gradient offers one of the best opportunities to study the interplay between natural selection and migration. In cases in which specific genetic variants also show such clinal patterns, it may be possible to uncover the mutations responsible for local adaptation. The malaria vector, Anopheles gambiae, is associated with a latitudinal cline in aridity in Cameroon; a large inversion on chromosome 2L of this mosquito ...

  2. Biochemical Characterization of Anopheles gambiae SRPN6, a Malaria Parasite Invasion Marker in Mosquitoes

    OpenAIRE

    Chunju An; Yasuaki Hiromasa; Xin Zhang; Scott Lovell; Michal Zolkiewski; John M Tomich; Kristin Michel

    2012-01-01

    Serine proteinase inhibitors of the serpin family are well known as negative regulators of hemostasis, thrombolysis and innate immune responses. Additionally, non-inhibitory serpins serve functions as chaperones, hormone transporters, or anti-angiogenic factors. In the African malaria mosquito, Anopheles gambiae s.s., at least three serpins (SRPNs) are implicated in the innate immune response against malaria parasites. Based on reverse genetic and cell biological analyses, AgSRPN6 limits para...

  3. The JNK Pathway Is a Key Mediator of Anopheles gambiae Antiplasmodial Immunity

    OpenAIRE

    Garver, Lindsey S.; de Almeida Oliveira, Giselle; Barillas-Mury, Carolina

    2013-01-01

    The innate immune system of Anopheles gambiae mosquitoes limits Plasmodium infection through multiple molecular mechanisms. For example, midgut invasion by the parasite triggers an epithelial nitration response that promotes activation of the complement-like system. We found that suppression of the JNK pathway, by silencing either Hep, JNK, Jun or Fos expression, greatly enhanced Plasmodium infection; while overactivating this cascade, by silencing the suppressor Puckered, had the opposite ef...

  4. A mosquito lipoxin/lipocalin complex mediates innate immune priming in Anopheles gambiae

    OpenAIRE

    Ramirez, Jose Luis; de Almeida Oliveira, Giselle; Calvo, Eric; Dalli, Jesmond; Colas, Romain A.; Serhan, Charles N.; Ribeiro, Jose M.; Barillas-Mury, Carolina

    2015-01-01

    Exposure of Anopheles gambiae mosquitoes to Plasmodium infection enhances the ability of their immune system to respond to subsequent infections. However, the molecular mechanism that allows the insect innate immune system to ‘remember' a previous encounter with a pathogen has not been established. Challenged mosquitoes constitutively release a soluble haemocyte differentiation factor into their haemolymph that, when transferred into Naive mosquitoes, also induces priming. Here we show that t...

  5. A Peroxidase/Dual Oxidase System Modulates Midgut Epithelial Immunity in Anopheles gambiae

    OpenAIRE

    Kumar, Sanjeev; Molina-Cruz, Alvaro; Gupta, Lalita; Rodrigues, Janneth; Barillas-Mury, Carolina

    2010-01-01

    Extracellular matrices in diverse biological systems are crosslinked by dityrosine covalent bonds catalyzed by the peroxidase/oxidase system. We show that the Immunomodulatory Peroxidase (IMPer), an enzyme secreted by the mosquito Anopheles gambiae midgut, and dual oxidase (Duox) form a dityrosine network that decreases gut permeability to immune elicitors and protects the microbiota by preventing activation of epithelial immunity. It also provides a suitable environment for malaria parasites...

  6. The STAT pathway mediates late phase immunity against Plasmodium in the mosquito Anopheles gambiae

    OpenAIRE

    Gupta, Lalita; Molina-Cruz, Alvaro; Kumar, Sanjeev; Rodrigues, Janneth; Dixit, Rajnikant; Zamora, Rodolfo E.; Barillas-Mury, Carolina

    2009-01-01

    The STAT family of transcription factors activate expression of immune system genes in vertebrates. The ancestral STAT gene (AgSTAT-A) appears to have duplicated in the mosquito Anopheles gambiae, giving rise to a second intronless STAT gene (AgSTAT-B), which we show regulates AgSTAT-A expression in adult females. AgSTAT-A participates in the transcriptional activation of nitric oxide synthase (NOS) in response to bacterial and plasmodial infection. Activation of this pathway, however, is not...

  7. Genome expression analysis of Anopheles gambiae: Responses to injury, bacterial challenge, and malaria infection

    OpenAIRE

    Dimopoulos, George; Christophides, George K.; Meister, Stephan; SCHULTZ, JÖRG; White, Kevin P.; Barillas-Mury, Carolina; Kafatos, Fotis C.

    2002-01-01

    The complex gene expression responses of Anopheles gambiae to microbial and malaria challenges, injury, and oxidative stress (in the mosquito and/or a cultured cell line) were surveyed by using cDNA microarrays constructed from an EST-clone collection. The expression profiles were broadly subdivided into induced and down-regulated gene clusters. Gram+ and Gram− bacteria and microbial elicitors up-regulated a diverse set of genes, many belonging to the immunity class, and the response to malar...

  8. Cloning and molecular characterization of two invertebrate-type lysozymes from Anopheles gambiae

    OpenAIRE

    Paskewitz, S M; Li, B.; Kajla, M. K.

    2008-01-01

    We sequenced and characterized two novel invertebrate-type lysozymes from the mosquito Anopheles gambiae. Alignment and phylogenetic analysis of these and a number of related insect proteins identified through bioinformatics strategies showed a high degree of conservation of this protein family throughout the Class Insecta. Expression profiles were examined for the two mosquito genes through semiquantitative and real-time PCR analysis. Lys i-1 transcripts were found in adult females in the fa...

  9. The genetics of inviability and male sterility in hybrids between Anopheles gambiae and An. arabiensis.

    Science.gov (United States)

    Slotman, M; Della Torre, A; Powell, J R

    2004-05-01

    Male hybrids between Anopheles gambiae and An. arabiensis suffer from hybrid sterility, and inviability effects are sometimes present as well. We examined the genetic basis of these reproductive barriers between the two species, using 21 microsatellite markers. Generally, recessive inviability effects were found on the X chromosome of gambiae that are incompatible with at least one factor on each arabiensis autosome. Inviability is complete when the gambiae and arabiensis inviability factors are hemi- or homozygous. Using a QTL mapping approach, regions that contribute to male hybrid sterility were also identified. The X chromosome has a disproportionately large effect on male hybrid sterility. Additionally, several moderate-to-large autosomal QTL were found in both species. The effect of these autosomal QTL is contingent upon the presence of an X chromosome from the other species. Substantial regions of the autosomes do not contribute markedly to male hybrid sterility. Finally, no evidence for epistatic interactions between conspecific sterility loci was found.

  10. New selenoproteins identified in silico from the genome of Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    JIANG Liang; LIU Qiong; CHEN Ping; GAO ZhongHong; XU HuiBi

    2007-01-01

    Selenoprotein is biosynthesized by the incorporation of selenocysteine into proteins, where the TGA codon in the open reading frame does not act as a stop signal but is translated into selenocysteine. The dual functions of TGA result in mis-annotation or lack of selenoproteins in the sequenced genomes of many species. Available computational tools fail to correctly predict selenoproteins. Thus, we developed a new method to identify selenoproteins from the genome of Anopheles gambiae computationally.Based on released genomic information, several programs were edited with PERL language to identify selenocysteine insertion sequence (SECIS) element, the coding potential of TGA codons, and cysteine-containing homologs of selenoprotein genes. Our results showed that 11365 genes were terminated with TGA codons, 918 of which contained SECIS elements. Similarity search revealed that 58genes contained Sec/Cys pairs and similar flanking regions around in-frame TGA codons. Finally, 7genes were found to fully meet requirements for selenoproteins, although they have not been annotated as selenoproteins in NCBI databases. Deduced from their basic properties, the newly found selenoproteins in the genome of Anopheles gambiae are possibly related to in vivo oxidation tolerance and protein regulation in order to interfere with anopheles' vectorial capacity of Plasmodium. This study may also provide theoretical bases for the prevention of malaria from anopheles transmission.

  11. Laboratory studies on the olfactory behaviour of Anopheles quadriannulatus

    NARCIS (Netherlands)

    Pates, H.V.; Takken, W.; Curtis, C.F.

    2005-01-01

    The host preference of Anopheles quadriannulatus Theobald (Diptera: Culicidae), the zoophilic member of the malaria mosquito complex Anopheles gambiae Giles, was investigated in a dual-choice olfactometer. Naïve female mosquitoes were exposed to CO2, acetone, 1-octen-3-ol, and skin emanations from c

  12. Islands and Stepping-Stones: Comparative Population Structure of Anopheles gambiae sensu stricto and Anopheles arabiensis in Tanzania and Implications for the Spread of Insecticide Resistance.

    OpenAIRE

    Deodatus Maliti; Hilary Ranson; Stephen Magesa; William Kisinza; Juma Mcha; Khamis Haji; Gerald Killeen; David Weetman

    2014-01-01

    Population genetic structures of the two major malaria vectors Anopheles gambiae s.s. and An. arabiensis, differ markedly across Sub-Saharan Africa, which could reflect differences in historical demographies or in contemporary gene flow. Elucidation of the degree and cause of population structure is important for predicting the spread of genetic traits such as insecticide resistance genes or artificially engineered genes. Here the population genetics of An. gambiae s.s. and An. arabiensis in ...

  13. Kdr-based insecticide resistance in Anopheles gambiae s.s populations in

    Directory of Open Access Journals (Sweden)

    Nwane Philippe

    2011-10-01

    Full Text Available Abstract Background The spread of insecticide resistance in the malaria mosquito, Anopheles gambiae is a serious threat for current vector control strategies which rely on the use of insecticides. Two mutations at position 1014 of the S6 transmembrane segment of domain II in the voltage gated sodium channel, known as kdr (knockdown resistance mutations leading to a change of a Leucine to a Phenylalanine (L1014F or to a Serine (L1014S confer resistance to DDT and pyrethroid insecticides in the insect. This paper presents the current distribution of the kdr alleles in wild Anopheles gambiae populations in Cameroon. Results A total of 1,405 anopheline mosquitoes were collected from 21 localities throughout Cameroon and identified as An. gambiae (N = 1,248; 88.8%, An. arabiensis (N = 120; 8.5% and An. melas (N = 37; 2.6%. Both kdr alleles 1014F and 1014S were identified in the M and S molecular forms of An. gambiae s.s. The frequency of the 1014F allele ranged from 1.7 to 18% in the M-form, and from 2 to 90% in the S-form. The 1014S allele ranged from 3-15% in the S-form and in the M-form its value was below 3%. Some specimens were found to carry both resistant kdr alleles. Conclusion This study provides an updated distribution map of the kdr alleles in wild An. gambiae populations in Cameroon. The co-occurrence of both alleles in malaria mosquito vectors in diverse ecological zones of the country may be critical for the planning and implementation of malaria vector control interventions based on IRS and ITNs, as currently ongoing in Cameroon.

  14. Bendiocarb, a potential alternative against pyrethroid resistant Anopheles gambiae in Benin, West Africa

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    Irish Seth

    2010-07-01

    Full Text Available Abstract Background Anopheles gambiae, the main malaria vector in Benin has developed high level of resistance to pyrethroid insecticides, which is a serious concern to the future use of long-lasting insecticidal nets (LLIN and indoor residual spraying (IRS. In this context, one of the pathways available for malaria vector control would be to investigate alternative classes of insecticides with different mode of action than that of pyrethroids. The goal of this study was to evaluate under field conditions the efficacy of a carbamate (bendiocarb and an organophosphate (fenitrothion against pyrethroid-resistant An. gambiae s.s. Methods Wild populations and females from laboratory colonies of five days old An. gambiae were bio-assayed during this study. Two pyrethroids (deltamethrin and alphacypermethrin, an organophosphate (fenitrothion, a carbamate (bendiocarb and a mixture of an organophosphate (chlorpyriphos + a pyrethroid deltamethrin were compared in experimental huts as IRS treatments. Insecticides were applied in the huts using a hand-operated compression sprayer. The deterrency, exophily, blood feeding rate and mortality induced by these insecticides against An. gambiae were compared to the untreated control huts. Results Deltamethrin, alphacypermethrin and bendiocarb treatment significantly reduced mosquito entry into the huts (p An. gambiae (in the first month and 77.8% (in the fourth month. Bendiocarb and the mixture chlorpyriphos/deltamethrin mortality rates ranged from 97.9 to 100% the first month and 77.7-88% the third month respectively. Conclusion After four months, fenitrothion, bendiocarb and the mixture chlorpyriphos/deltamethrin performed effectively against pyrethroid-resistant Anopheles. These results showed that bendiocarb could be recommended as an effective insecticide for use in IRS operations in Benin, particularly as the mixture chlorpyriphos/deltamethrin does not have WHOPES authorization and complaints were mentioned

  15. Physiology and development of the M and S molecular forms of Anopheles gambiae in Burkina Faso (West Africa).

    Science.gov (United States)

    Mouline, K; Mamai, W; Agnew, P; Tchonfienet, M; Brengues, C; Dabire, R; Robert, V; Simard, F

    2012-12-01

    In West Africa, M and S molecular forms of Anopheles gambiae sensu stricto (Diptera: Culicidae) Giles, frequently occur together, although with different population bionomics. The S form typically breeds in rain-dependant water collections and is present during the rainy season only whereas the M form can thrive all year long in areas with permanent breeding opportunities. In the present study, we explored physiological and developmental trade-offs at play in laboratory colonies and field populations of the M and S forms that originated from an area of sympatry in Burkina Faso, where M and S larvae exhibit such habitat segregation. In the laboratory, larvae of the M form developed slower than the S form (mean values 9.51 and 8.85 days, respectively, Wilcoxon's test, P < 0.001). Although wing length and dry weight at emergence showed large variations, M females were on average 8% heavier than S females of similar wing length. Higher nutritional reserves (proteins and lipids) in teneral adults explained part of this weight difference, reflecting a better ability of the M form to garner resources at the larval stage. Furthermore, a higher rate of ovarian maturation was observed in the M form after a single bloodmeal. The relevance of these findings for parasite transmission is discussed. PMID:22681446

  16. A comprehensive gene expression atlas of sex- and tissue-specificity in the malaria vector, Anopheles gambiae

    Directory of Open Access Journals (Sweden)

    Crisanti Andrea

    2011-06-01

    Full Text Available Abstract Background The mosquito, Anopheles gambiae, is the primary vector of human malaria, a disease responsible for millions of deaths each year. To improve strategies for controlling transmission of the causative parasite, Plasmodium falciparum, we require a thorough understanding of the developmental mechanisms, physiological processes and evolutionary pressures affecting life-history traits in the mosquito. Identifying genes expressed in particular tissues or involved in specific biological processes is an essential part of this process. Results In this study, we present transcription profiles for ~82% of annotated Anopheles genes in dissected adult male and female tissues. The sensitivity afforded by examining dissected tissues found gene activity in an additional 20% of the genome that is undetected when using whole-animal samples. The somatic and reproductive tissues we examined each displayed patterns of sexually dimorphic and tissue-specific expression. By comparing expression profiles with Drosophila melanogaster we also assessed which genes are well conserved within the Diptera versus those that are more recently evolved. Conclusions Our expression atlas and associated publicly available database, the MozAtlas (http://www.tissue-atlas.org, provides information on the relative strength and specificity of gene expression in several somatic and reproductive tissues, isolated from a single strain grown under uniform conditions. The data will serve as a reference for other mosquito researchers by providing a simple method for identifying where genes are expressed in the adult, however, in addition our resource will also provide insights into the evolutionary diversity associated with gene expression levels among species.

  17. Spermless males elicit large-scale female responses to mating in the malaria mosquito Anopheles gambiae

    Science.gov (United States)

    Thailayil, Janis; Magnusson, Kalle; Godfray, H. Charles J.; Crisanti, Andrea; Catteruccia, Flaminia

    2011-01-01

    Anopheles gambiae sensu stricto is the major vector of malaria, a disease with devastating consequences for human health. Given the constant spread of the disease, alternative approaches to the use of insecticides are urgently needed to control vector populations. Females of this species undergo large behavioral changes after mating, which include a life-long refractoriness to further insemination and the induction of egg laying in blood-fed individuals. Genetic control strategies aimed at impacting Anopheles fertility through the release of sterile males are being advocated to reduce the size of mosquito field populations. Such strategies depend on the ability of the released sterile males to mate successfully with wild females and to switch off the female receptivity to further copulation. Here we evaluate the role of sperm in regulating female behavioral responses after mating in An. gambiae. We developed spermless males by RNAi silencing of a germ cell differentiation gene. These males mated successfully and preserved standard accessory gland functions. Females mated to spermless males exhibited normal postcopulatory responses, which included laying large numbers of eggs upon blood feeding and becoming refractory to subsequent insemination. Moreover, spermless males induced transcriptional changes in female reproductive genes comparable to those elicited by fertile males. Our data demonstrate that, in contrast to Drosophila, targeting sperm in An. gambiae preserves normal male and female reproductive behavior for the traits and time frame analyzed and validate the use of approaches based on incapacitation or elimination of sperm for genetic control of vector populations to block malaria transmission. PMID:21825136

  18. New selenoproteins identified in silico from the genome of Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    2007-01-01

    Selenoprotein is biosynthesized by the incorporation of selenocysteine into proteins,where the TGA codon in the open reading frame does not act as a stop signal but is translated into selenocysteine.The dual functions of TGA result in mis-annotation or lack of selenoproteins in the sequenced genomes of many species.Available computational tools fail to correctly predict selenoproteins.Thus,we devel-oped a new method to identify selenoproteins from the genome of Anopheles gambiae computationally.Based on released genomic information,several programs were edited with PERL language to identify selenocysteine insertion sequence(SECIS)element,the coding potential of TGA codons,and cys-teine-containing homologs of selenoprotein genes.Our results showed that 11365 genes were termi-nated with TGA codons,918 of which contained SECIS elements.Similarity search revealed that 58 genes contained Sec/Cys pairs and similar flanking regions around in-frame TGA codons.Finally,7 genes were found to fully meet requirements for selenoproteins,although they have not been anno-tated as selenoproteins in NCBI databases.Deduced from their basic properties,the newly found se-lenoproteins in the genome of Anopheles gambiae are possibly related to in vivo oxidation tolerance and protein regulation in order to interfere with anopheles’ vectorial capacity of Plasmodium.This study may also provide theoretical bases for the prevention of malaria from anopheles transmission.

  19. Insecticide resistance in Anopheles gambiae from south-western Chad, Central Africa

    Directory of Open Access Journals (Sweden)

    Etang Josiane

    2008-09-01

    Full Text Available Abstract Background Indoor residual spraying and insecticide-treated nets (ITN are essential components of malaria vector control in Africa. Pyrethroids are the only recommended compounds for nets treatment because they are fast-acting insecticides with low mammalian toxicity. However, there is growing concern that pyrethroid resistance may threaten the sustainability of ITN scaling-up programmes. Here, insecticide susceptibility was investigated in Anopheles gambiae sensu lato from an area of large scale ITN distribution programme in south-western Chad. Methods Susceptibility to 4% DDT, 0.05% deltamethrin, 0.75% permethrin, 0.1% bendiocarb and 5% malathion was assessed using the WHO standard procedures for adult mosquitoes. Tests were carried out with two to four days-old, non-engorged female mosquitoes. The An. gambiae Kisumu strain was used as a reference. Knockdown effect was recorded every 5 min and mortality scored 24 h after exposure. Mosquitoes were identified to species and molecular form by PCR-RFLP and genotypes at the kdr locus were determined in surviving specimens by Hot Oligonucleotide Ligation Assay (HOLA. Results During this survey, full susceptibility to malathion was recorded in all samples. Reduced susceptibility to bendiocarb (mortality rate of 96.1% was found in one sample out of nine assayed. Increased tolerance to pyrethroids was detected in most samples (8/9 with mortality rates ranging from 70.2 to 96.6% for deltamethrin and from 26.7 to 96.3% for permethrin. Pyrethroid tolerance was not associated with a significant increase of knock-down times. Anopheles arabiensis was the predominant species of the An. gambiae complex in the study area, representing 75 to 100% of the samples. Screening for kdr mutations detected the L1014F mutation in 88.6% (N = 35 of surviving An. gambiae sensu stricto S form mosquitoes. All surviving An. arabiensis (N = 49 and M form An. gambiae s.s. (N = 1 carried the susceptible allele

  20. Life on the edge: African malaria mosquito (Anopheles gambiae s. l.) larvae are amphibious

    Science.gov (United States)

    Miller, James R.; Huang, Juan; Vulule, John; Walker, Edward D.

    2007-03-01

    Anopheles gambiae s.l. is the main vector of malaria in Sub-Saharan Africa. Here, an estimated 1 million people die every year from this disease. Despite considerable research on An. gambiae that increasingly explores sub-organismal phenomena, important facets of the field biology of this deadly insect are yet being discovered. In the current study, we used simple observational tools to reveal that the habitat of larval An. gambiae is not limited within the boundaries of temporary mud puddles, as has been the accepted generalization. Thus, control tactics aimed at immatures must consider zones larger than puddles per se. In fact, eggs are more likely to be found outside than inside puddles. Eggs can develop and larvae can emerge on mud. Larvae are then capable of three distinct modes of terrestrial displacement (two active and one passive), whereby, they can reach standing water. On mud bearing a film of water, larvae actively displace backwards by sinusoidal undulations shown to be only a slight variation of the swimming motor program. On drying mud, larvae switch to a slower and forward form of active locomotion resembling that of a crawling caterpillar. During rains, small larvae may be passively displaced by flowing rainwater so as to be deposited into puddles. These capabilities for being amphibious, along with very rapid growth and development, help explain how An. gambiae thrives in a highly uncertain and often hostile larval environment.

  1. Anopheles gambiae eicosanoids modulate Plasmodium berghei survival from oocyst to salivary gland invasion

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    Susana Ramos

    2014-08-01

    Full Text Available Eicosanoids affect the immunity of several pathogen/insect models, but their role on the Anopheles gambiae response to Plasmodium is still unknown. Plasmodium berghei-infected mosquitoes were injected with an eicosanoid biosynthesis inhibitor, indomethacin (IN, or a substrate, arachidonic acid (AA, at day 7 or day 12 post-infection (p.i.. Salivary gland invasion was evaluated by sporozoite counts at day 21 p.i. IN promoted infection upon sporozoite release from oocysts, but inhibited infection when sporozoites were still maturing within the oocysts, as observed by a reduction in the number of sporozoites reaching the salivary glands. AA treatment had the opposite effect. We show for the first time that An. gambiae can modulate parasite survival through eicosanoids by exerting an antagonistic or agonistic effect on the parasite, depending on its stage of development.

  2. Energy metabolism affects susceptibility of Anopheles gambiae mosquitoes to Plasmodium infection.

    Science.gov (United States)

    Oliveira, Jose Henrique M; Gonçalves, Renata L S; Oliveira, Giselle A; Oliveira, Pedro L; Oliveira, Marcus F; Barillas-Mury, Carolina

    2011-06-01

    Previous studies showed that Anopheles gambiae L3-5 females, which are refractory (R) to Plasmodium infection, express higher levels of genes involved in redox-metabolism and mitochondrial respiration than susceptible (S) G3 females. Our studies revealed that R females have reduced longevity, faster utilization of lipid reserves, impaired mitochondrial state-3 respiration, increased rate of mitochondrial electron leak and higher expression levels of several glycolytic enzyme genes. Furthermore, when state-3 respiration was reduced in S females by silencing expression of the adenine nucleotide translocator (ANT), hydrogen peroxide generation was higher and the mRNA levels of lactate dehydrogenase increased in the midgut, while the prevalence and intensity of Plasmodium berghei infection were significantly reduced. We conclude that there are broad metabolic differences between R and S An. gambiae mosquitoes that influence their susceptibility to Plasmodium infection. PMID:21320598

  3. Polymorphisms in Anopheles gambiae immune genes associated with natural resistance to Plasmodium falciparum.

    Directory of Open Access Journals (Sweden)

    Caroline Harris

    Full Text Available Many genes involved in the immune response of Anopheles gambiae, the main malaria vector in Africa, have been identified, but whether naturally occurring polymorphisms in these genes underlie variation in resistance to the human malaria parasite, Plasmodium falciparum, is currently unknown. Here we carried out a candidate gene association study to identify single nucleotide polymorphisms (SNPs associated with natural resistance to P. falciparum. A. gambiae M form mosquitoes from Cameroon were experimentally challenged with three local wild P. falciparum isolates. Statistical associations were assessed between 157 SNPs selected from a set of 67 A. gambiae immune-related genes and the level of infection. Isolate-specific associations were accounted for by including the effect of the isolate in the analysis. Five SNPs were significantly associated to the infection phenotype, located within or upstream of AgMDL1, CEC1, Sp PPO activate, Sp SNAKElike, and TOLL6. Low overall and local linkage disequilibrium indicated high specificity in the loci found. Association between infection phenotype and two SNPs was isolate-specific, providing the first evidence of vector genotype by parasite isolate interactions at the molecular level. Four SNPs were associated to either oocyst presence or load, indicating that the genetic basis of infection prevalence and intensity may differ. The validity of the approach was verified by confirming the functional role of Sp SNAKElike in gene silencing assays. These results strongly support the role of genetic variation within or near these five A. gambiae immune genes, in concert with other genes, in natural resistance to P. falciparum. They emphasize the need to distinguish between infection prevalence and intensity and to account for the genetic specificity of vector-parasite interactions in dissecting the genetic basis of Anopheles resistance to human malaria.

  4. A simplified high-throughput method for pyrethroid knock-down resistance (kdr) detection in Anopheles gambiae

    OpenAIRE

    Walker Edward D; Black William C; Randle Nadine P; McCall P J; Ranson Hilary; Lynd Amy; Donnelly Martin J

    2005-01-01

    Abstract Background A single base pair mutation in the sodium channel confers knock-down resistance to pyrethroids in many insect species. Its occurrence in Anopheles mosquitoes may have important implications for malaria vector control especially considering the current trend for large scale pyrethroid-treated bednet programmes. Screening Anopheles gambiae populations for the kdr mutation has become one of the mainstays of programmes that monitor the development of insecticide resistance. Th...

  5. Larvicidal activity of extracts from three Plumbago spp against Anopheles gambiae

    OpenAIRE

    Barasa M Maniafu; Lwande Wilber; Ndiege, Isaiah O.; Cornelius C Wanjala; Teresa Ayuko Akenga

    2009-01-01

    Three Plumbago spp have been tested for mosquito larvicidal activity. The crude extracts exhibiting the highest larvicidal activity against Anopheles gambiae were hexane (LC50 = 6.4 μg/mL) and chloroform (LC50 = 6.7 μg/mL) extracts from Plumbago zeylanica Linn, chloroform (LC50 = 6.7 ug/mL) extract from Plumbago stenophylla Bull and ethyl acetate (LC50 = 4.1 μg/mL) extract from Plumbago dawei Rolfe. These LC50 values were within 95% confidence limits. 5-hydroxy-2-methyl-1,4-nap...

  6. Effect of ivermectin on the larvae of Anopheles gambiae and Culex quinquefasciatus

    DEFF Research Database (Denmark)

    Derua, Yahya A.; Malongo, Bernard B.; Simonsen, Paul E.

    2016-01-01

    Background: Ivermectin is used extensively globally for treatment of helminthic and ectoparasitic infections in animals and humans. The effect of excreted ivermectin on non-target organisms in aquatic and terrestrial environments has been increasingly reported. Due to its low water solubility...... and adsorption to sediments, the ivermectin exposure-risk to aquatic organisms dwelling in different strata of water bodies varies. This study assessed the survival of larvae of Anopheles gambiae Giles and Culex quinquefasciatus Say, when exposed to low concentrations of ivermectin under laboratory conditions...

  7. A peroxidase/dual oxidase system modulates midgut epithelial immunity in Anopheles gambiae.

    Science.gov (United States)

    Kumar, Sanjeev; Molina-Cruz, Alvaro; Gupta, Lalita; Rodrigues, Janneth; Barillas-Mury, Carolina

    2010-03-26

    Extracellular matrices in diverse biological systems are cross-linked by dityrosine covalent bonds catalyzed by the peroxidase/oxidase system. We show that a peroxidase, secreted by the Anopheles gambiae midgut, and dual oxidase form a dityrosine network that decreases gut permeability to immune elicitors. This network protects the microbiota by preventing activation of epithelial immunity. It also provides a suitable environment for malaria parasites to develop within the midgut lumen without inducing nitric oxide synthase expression. Disruption of this barrier results in strong and effective pathogen-specific immune responses. PMID:20223948

  8. A Peroxidase/Dual Oxidase System Modulates Midgut Epithelial Immunity in Anopheles gambiae

    Science.gov (United States)

    Kumar, Sanjeev; Molina-Cruz, Alvaro; Gupta, Lalita; Rodrigues, Janneth; Barillas-Mury, Carolina

    2012-01-01

    Extracellular matrices in diverse biological systems are crosslinked by dityrosine covalent bonds catalyzed by the peroxidase/oxidase system. We show that the Immunomodulatory Peroxidase (IMPer), an enzyme secreted by the mosquito Anopheles gambiae midgut, and dual oxidase (Duox) form a dityrosine network that decreases gut permeability to immune elicitors and protects the microbiota by preventing activation of epithelial immunity. It also provides a suitable environment for malaria parasites to develop within the midgut lumen without inducing nitric oxide synthase expression. Disruption of this barrier results in strong and effective pathogen-specific immune responses. PMID:20223948

  9. Gene expression patterns associated with blood-feeding in the malaria mosquito Anopheles gambiae

    OpenAIRE

    Hogan James R; Lobo Neil F; Harker Brent W; Hillenmeyer Maureen E; Kern Marcia K; Hong Young S; Dana Ali N; Romans Patricia; Collins Frank H

    2005-01-01

    Abstract Background Blood feeding, or hematophagy, is a behavior exhibited by female mosquitoes required both for reproduction and for transmission of pathogens. We determined the expression patterns of 3,068 ESTs, representing ~2,000 unique gene transcripts using cDNA microarrays in adult female Anopheles gambiae at selected times during the first two days following blood ingestion, at 5 and 30 min during a 40 minute blood meal and at 0, 1, 3, 5, 12, 16, 24 and 48 hours after completion of t...

  10. The role of reactive oxygen species on Plasmodium melanotic encapsulation in Anopheles gambiae.

    Science.gov (United States)

    Kumar, Sanjeev; Christophides, George K; Cantera, Rafael; Charles, Bradley; Han, Yeon Soo; Meister, Stephan; Dimopoulos, George; Kafatos, Fotis C; Barillas-Mury, Carolina

    2003-11-25

    Malaria transmission depends on the competence of some Anopheles mosquitoes to sustain Plasmodium development (susceptibility). A genetically selected refractory strain of Anopheles gambiae blocks Plasmodium development, melanizing, and encapsulating the parasite in a reaction that begins with tyrosine oxidation, and involves three quantitative trait loci. Morphological and microarray mRNA expression analysis suggest that the refractory and susceptible strains have broad physiological differences, which are related to the production and detoxification of reactive oxygen species. Physiological studies corroborate that the refractory strain is in a chronic state of oxidative stress, which is exacerbated by blood feeding, resulting in increased steady-state levels of reactive oxygen species, which favor melanization of parasites as well as Sephadex beads. PMID:14623973

  11. Three years of insecticide resistance monitoring in Anopheles gambiae in Burkina Faso: resistance on the rise?

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    Badolo Athanase

    2012-07-01

    Full Text Available Abstract Background and methods A longitudinal Anopheles gambiae s.l. insecticide-resistance monitoring programme was established in four sentinel sites in Burkina Faso. For three years, between 2008 and 2010, WHO diagnostic dose assays were used to measure the prevalence of resistance to all the major classes of insecticides at the beginning and end of the malaria transmission season. Species identification and genotyping for target site mutations was also performed and the sporozoite rate in adults determined. Results At the onset of the study, resistance to DDT and pyrethroids was already prevalent in An. gambiae s.l. from the south-west of the country but mosquitoes from the two sites in central Burkina Faso were largely susceptible. Within three years, DDT and permethrin resistance was established in all four sites. Carbamate and organophosphate resistance remains relatively rare and largely confined to the south-western areas although a small number of bendiocarb survivors were found in all sites by the final round of monitoring. The ace-1R target site resistance allele was present in all localities and its frequency exceeded 20% in 2010 in two of the sites. The frequency of the 1014F kdr mutation increased throughout the three years and by 2010, the frequency of 1014F in all sites combined was 0.02 in Anopheles arabiensis, 0.56 in An. gambiae M form and 0.96 in An. gambiae S form. This frequency did not differ significantly between the sites. The 1014S kdr allele was only found in An. arabiensis but its frequency increased significantly throughout the study (P = 0.0003 and in 2010 the 1014S allele frequency was 0.08 in An. arabiensis. Maximum sporozoite rates (12% were observed in Soumousso in 2009 and the difference between sites is significant for each year. Conclusion Pyrethroid and DDT resistance is now established in An. gambiae s.l. throughout Burkina Faso. Results from diagnostic dose assays are highly variable within and

  12. Factors affecting fungus-induced larval mortality in Anopheles gambiae and Anopheles stephensi

    NARCIS (Netherlands)

    Bukhari, S.T.; Middelman, A.; Koenraadt, C.J.M.; Takken, W.; Knols, B.G.J.

    2010-01-01

    Background Entomopathogenic fungi have shown great potential for the control of adult malaria vectors. However, their ability to control aquatic stages of anopheline vectors remains largely unexplored. Therefore, how larval characteristics (Anopheles species, age and larval density), fungus (species

  13. Predation efficiency of Anopheles gambiae larvae by aquatic predators in western Kenya highlands

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    Nyindo Mramba

    2011-07-01

    Full Text Available Abstract Background The current status of insecticide resistance in mosquitoes and the effects of insecticides on non-target insect species have raised the need for alternative control methods for malaria vectors. Predation has been suggested as one of the important regulation mechanisms for malaria vectors in long-lasting aquatic habitats, but the predation efficiency of the potential predators is largely unknown in the highlands of western Kenya. In the current study, we examined the predation efficiency of five predators on Anopheles gambiae s.s larvae in 24 hour and semi- field evaluations. Methods Predators were collected from natural habitats and starved for 12 hours prior to starting experiments. Preliminary experiments were conducted to ascertain the larval stage most predated by each predator species. When each larval instar was subjected to predation, third instar larvae were predated at the highest rate. Third instar larvae of An. gambiae were introduced into artificial habitats with and without refugia at various larval densities. The numbers of surviving larvae were counted after 24 hours in 24. In semi-field experiments, the larvae were counted daily until they were all either consumed or had developed to the pupal stage. Polymerase chain reaction was used to confirm the presence of An. gambiae DNA in predator guts. Results Experiments found that habitat type (P P P P An. gambiae DNA was found in at least three out of ten midguts for all predator species. Gambusia affins was the most efficient, being three times more efficient than tadpoles. Conclusion These experiments provide insight into the efficiency of specific natural predators against mosquito larvae. These naturally occurring predators may be useful in biocontrol strategies for aquatic stage An. gambiae mosquitoes. Further investigations should be done in complex natural habitats for these predators.

  14. Differential attractiveness of humans to the African malaria vector Anopheles gambiae Giles - Effects of host characteristics and parasite infection

    NARCIS (Netherlands)

    Mukabana, W.R.

    2002-01-01

    The results of a series of studies designed to understand the principal factors that determine the differential attractiveness of humans to the malaria vector Anopheles gambiae are described in this thesis. Specific

  15. Toxicity of six plant extracts and two pyridine alkaloids from Ricinus communis against the malaria vector Anopheles gambiae

    Science.gov (United States)

    The African malaria vector, Anopheles gambiae s.s., is known to feed selectively on certain plants for sugar sources. However, the adaptive significance of this behavior especially on how the extracts of such plants impact on the fitness of this vector has not been explored. This study determined th...

  16. Comparative evaluation of systemic drugs for their effects against Anopheles gambiae.

    Science.gov (United States)

    Butters, Matthew P; Kobylinski, Kevin C; Deus, Kelsey M; da Silva, Ines Marques; Gray, Meg; Sylla, Massamba; Foy, Brian D

    2012-01-01

    Laboratory and field studies have shown that ivermectin, a drug that targets invertebrate ligand-gated ion channels (LGICs), is potently active against Anopheles spp. mosquitoes at concentrations present in human blood after standard drug administrations; thus ivermectin holds promise as a mass human-administered endectocide that could help suppress malaria parasite transmission. We evaluated other systemic LGIC-targeting drugs for their activities against the African malaria vector Anopheles gambiae using in vitro blood feeding assays. Eprinomectin, selamectin, moxidectin, and N-tert-butyl nodulisporamide were evaluated as potentially systemic drugs having similar modes of action to ivermectin; all primarily are agonists of invertebrate glutamate-gated chloride ion channels. Additionally, nitenpyram and spinosad were evaluated as systemic drugs that primarily work as agonists of nicotinic acetylcholine receptor channels. Only eprinomectin killed An. gambiae at concentrations that were comparable to ivermectin. At sub-lethal doses, nitenpyram and moxidectin marginally affected mosquito re-blood feeding ability. The macrocyclic lactones, particularly eprinomectin, caused significantly increased knockdown and significantly inhibited recovery in blood fed females. These data are a first step in evaluating drugs that might be eventually combined with, or substituted for ivermectin for future malaria parasite transmission control. PMID:22019935

  17. Maternal environment shapes the life history and susceptibility to malaria of Anopheles gambiae mosquitoes

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    Lorenz Lena M

    2011-12-01

    Full Text Available Abstract Background It is becoming generally recognized that an individual's phenotype can be shaped not only by its own genotype and environmental experience, but also by its mother's environment and condition. Maternal environmental factors can influence mosquitoes' population dynamics and susceptibility to malaria, and therefore directly and indirectly the epidemiology of malaria. Methods In a full factorial experiment, the effects of two environmental stressors - food availability and infection with the microsporidian parasite Vavraia culicis - of female mosquitoes (Anopheles gambiae sensu stricto on their offspring's development, survival and susceptibility to malaria were studied. Results The offspring of A. gambiae s.s. mothers infected with V. culicis developed into adults more slowly than those of uninfected mothers. This effect was exacerbated when mothers were reared on low food. Maternal food availability had no effect on the survival of their offspring up to emergence, and microsporidian infection decreased survival only slightly. Low food availability for mothers increased and V. culicis-infection of mothers decreased the likelihood that the offspring fed on malaria-infected blood harboured malaria parasites (but neither maternal treatment influenced their survival up to dissection. Conclusions Resource availability and infection with V. culicis of A. gambiae s.s. mosquitoes not only acted as direct environmental stimuli for changes in the success of one generation, but could also lead to maternal effects. Maternal V. culicis infection could make offspring more resistant and less likely to transmit malaria, thus enhancing the efficacy of the microsporidian for the biological control of malaria.

  18. Larvicidal effects of a neem (Azadirachta indica oil formulation on the malaria vector Anopheles gambiae

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    Knols Bart GJ

    2007-05-01

    Full Text Available Abstract Background Larviciding is a key strategy used in many vector control programmes around the world. Costs could be reduced if larvicides could be manufactured locally. The potential of natural products as larvicides against the main African malaria vector, Anopheles gambiae s.s was evaluated. Methods To assess the larvicidal efficacy of a neem (Azadirachta indica oil formulation (azadirachtin content of 0.03% w/v on An. gambiae s.s., larvae were exposed as third and fourth instars to a normal diet supplemented with the neem oil formulations in different concentrations. A control group of larvae was exposed to a corn oil formulation in similar concentrations. Results Neem oil had an LC50 value of 11 ppm after 8 days, which was nearly five times more toxic than the corn oil formulation. Adult emergence was inhibited by 50% at a concentration of 6 ppm. Significant reductions on growth indices and pupation, besides prolonged larval periods, were observed at neem oil concentrations above 8 ppm. The corn oil formulation, in contrast, produced no growth disruption within the tested range of concentrations. Conclusion Neem oil has good larvicidal properties for An. gambiae s.s. and suppresses successful adult emergence at very low concentrations. Considering the wide distribution and availability of this tree and its products along the East African coast, this may prove a readily available and cheap alternative to conventional larvicides.

  19. Modelling Anopheles gambiae s.s. Population Dynamics with Temperature- and Age-Dependent Survival

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    Céline Christiansen-Jucht

    2015-05-01

    Full Text Available Climate change and global warming are emerging as important threats to human health, particularly through the potential increase in vector- and water-borne diseases. Environmental variables are known to affect substantially the population dynamics and abundance of the poikilothermic vectors of disease, but the exact extent of this sensitivity is not well established. Focusing on malaria and its main vector in Africa, Anopheles gambiae sensu stricto, we present a set of novel mathematical models of climate-driven mosquito population dynamics motivated by experimental data suggesting that in An. gambiae, mortality is temperature and age dependent. We compared the performance of these models to that of a “standard” model ignoring age dependence. We used a longitudinal dataset of vector abundance over 36 months in sub-Saharan Africa for comparison between models that incorporate age dependence and one that does not, and observe that age-dependent models consistently fitted the data better than the reference model. This highlights that including age dependence in the vector component of mosquito-borne disease models may be important to predict more reliably disease transmission dynamics. Further data and studies are needed to enable improved fitting, leading to more accurate and informative model predictions for the An. gambiae malaria vector as well as for other disease vectors.

  20. Modelling Anopheles gambiae s.s. Population Dynamics with Temperature- and Age-Dependent Survival.

    Science.gov (United States)

    Christiansen-Jucht, Céline; Erguler, Kamil; Shek, Chee Yan; Basáñez, María-Gloria; Parham, Paul E

    2015-06-01

    Climate change and global warming are emerging as important threats to human health, particularly through the potential increase in vector- and water-borne diseases. Environmental variables are known to affect substantially the population dynamics and abundance of the poikilothermic vectors of disease, but the exact extent of this sensitivity is not well established. Focusing on malaria and its main vector in Africa, Anopheles gambiae sensu stricto, we present a set of novel mathematical models of climate-driven mosquito population dynamics motivated by experimental data suggesting that in An. gambiae, mortality is temperature and age dependent. We compared the performance of these models to that of a "standard" model ignoring age dependence. We used a longitudinal dataset of vector abundance over 36 months in sub-Saharan Africa for comparison between models that incorporate age dependence and one that does not, and observe that age-dependent models consistently fitted the data better than the reference model. This highlights that including age dependence in the vector component of mosquito-borne disease models may be important to predict more reliably disease transmission dynamics. Further data and studies are needed to enable improved fitting, leading to more accurate and informative model predictions for the An. gambiae malaria vector as well as for other disease vectors. PMID:26030468

  1. The population genomics of trans-specific inversion polymorphisms in Anopheles gambiae.

    Science.gov (United States)

    White, Bradley J; Cheng, Changde; Sangaré, Djibril; Lobo, Neil F; Collins, Frank H; Besansky, Nora J

    2009-09-01

    In the malaria mosquito Anopheles gambiae polymorphic chromosomal inversions may play an important role in adaptation to environmental variation. Recently, we used microarray-based divergence mapping combined with targeted resequencing to map nucleotide differentiation between alternative arrangements of the 2La inversion. Here, we applied the same technique to four different polymorphic inversions on the 2R chromosome of An. gambiae. Surprisingly, divergence was much lower between alternative arrangements for all 2R inversions when compared to the 2La inversion. For one of the rearrangements, 2Ru, we successfully mapped a very small region (approximately 100 kb) of elevated divergence. For the other three rearrangements, we did not identify any regions of significantly high divergence, despite ample independent evidence from natural populations of geographic clines and seasonal cycling, and stable heterotic polymorphisms in laboratory populations. If these inversions are the targets of selection as hypothesized, we suggest that divergence between rearrangements may have escaped detection due to retained ancestral polymorphism in the case of the youngest 2R rearrangements and to extensive gene flux in the older 2R inversion systems that segregate in both An. gambiae and its sibling species An. arabiensis. PMID:19581444

  2. Sexual transfer of the steroid hormone 20E induces the postmating switch in Anopheles gambiae

    Science.gov (United States)

    Gabrieli, Paolo; Kakani, Evdoxia G.; Mitchell, Sara N.; Mameli, Enzo; Want, Elizabeth J.; Mariezcurrena Anton, Ainhoa; Serrao, Aurelio; Baldini, Francesco; Catteruccia, Flaminia

    2014-01-01

    Female insects generally mate multiple times during their lives. A notable exception is the female malaria mosquito Anopheles gambiae, which after sex loses her susceptibility to further copulation. Sex in this species also renders females competent to lay eggs developed after blood feeding. Despite intense research efforts, the identity of the molecular triggers that cause the postmating switch in females, inducing a permanent refractoriness to further mating and triggering egg-laying, remains elusive. Here we show that the male-transferred steroid hormone 20-hydroxyecdysone (20E) is a key regulator of monandry and oviposition in An. gambiae. When sexual transfer of 20E is impaired by partial inactivation of the hormone and inhibition of its biosynthesis in males, oviposition and refractoriness to further mating in the female are strongly reduced. Conversely, mimicking sexual delivery by injecting 20E into virgin females switches them to an artificial mated status, triggering egg-laying and reducing susceptibility to copulation. Sexual transfer of 20E appears to incapacitate females physically from receiving seminal fluids by a second male. Comparative analysis of microarray data from females after mating and after 20E treatment indicates that 20E-regulated molecular pathways likely are implicated in the postmating switch, including cytoskeleton and musculature-associated genes that may render the atrium impenetrable to additional mates. By revealing signals and pathways shaping key processes in the An. gambiae reproductive biology, our data offer new opportunities for the control of natural populations of malaria vectors. PMID:25368171

  3. Immune factor Gambif1, a new rel family member from the human malaria vector, Anopheles gambiae.

    Science.gov (United States)

    Barillas-Mury, C; Charlesworth, A; Gross, I; Richman, A; Hoffmann, J A; Kafatos, F C

    1996-09-01

    A novel rel family member, Gambif1 (gambiae immune factor 1), has been cloned from the human malaria vector, Anopheles gambiae, and shown to be most similar to Drosophila Dorsal and Dif. Gambif1 protein is translocated to the nucleus in fat body cells in response to bacterial challenge, although the mRNA is present at low levels at all developmental stages and is not induced by infection. DNA binding activity to the kappaB-like sites in the A.gambiae Defensin and the Drosophila Diptericin and Cecropin promoters is also induced in larval nuclear extracts following infection. Gambif1 has the ability to bind to kappaB-like sites in vitro. Co-transfection assays in Drosophila mbn-2 cells show that Gambif1 can activate transcription by interacting with the Drosophila Diptericin regulatory elements, but is not functionally equivalent to Dorsal in this assay. Gambif1 protein translocation to the nucleus and the appearance of kappaB-like DNA binding activity can serve as molecular markers of activation of the immune system and open up the possibility of studying the role of defence reactions in determining mosquito susceptibility/refractoriness to malaria infection. PMID:8887560

  4. The JNK pathway is a key mediator of Anopheles gambiae antiplasmodial immunity.

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    Lindsey S Garver

    Full Text Available The innate immune system of Anopheles gambiae mosquitoes limits Plasmodium infection through multiple molecular mechanisms. For example, midgut invasion by the parasite triggers an epithelial nitration response that promotes activation of the complement-like system. We found that suppression of the JNK pathway, by silencing either Hep, JNK, Jun or Fos expression, greatly enhanced Plasmodium infection; while overactivating this cascade, by silencing the suppressor Puckered, had the opposite effect. The JNK pathway limits infection via two coordinated responses. It induces the expression of two enzymes (HPx2 and NOX5 that potentiate midgut epithelial nitration in response to Plasmodium infection and regulates expression of two key hemocyte-derived immune effectors (TEP1 and FBN9. Furthermore, the An. gambiae L3-5 strain that has been genetically selected to be refractory (R to Plasmodium infection exhibits constitutive overexpression of genes from the JNK pathway, as well as midgut and hemocyte effector genes. Silencing experiments confirmed that this cascade mediates, to a large extent, the drastic parasite elimination phenotype characteristic of this mosquito strain. In sum, these studies revealed the JNK pathway as a key regulator of the ability of An. gambiae mosquitoes to limit Plasmodium infection and identified several effector genes mediating these responses.

  5. The JNK pathway is a key mediator of Anopheles gambiae antiplasmodial immunity.

    Science.gov (United States)

    Garver, Lindsey S; de Almeida Oliveira, Giselle; Barillas-Mury, Carolina

    2013-01-01

    The innate immune system of Anopheles gambiae mosquitoes limits Plasmodium infection through multiple molecular mechanisms. For example, midgut invasion by the parasite triggers an epithelial nitration response that promotes activation of the complement-like system. We found that suppression of the JNK pathway, by silencing either Hep, JNK, Jun or Fos expression, greatly enhanced Plasmodium infection; while overactivating this cascade, by silencing the suppressor Puckered, had the opposite effect. The JNK pathway limits infection via two coordinated responses. It induces the expression of two enzymes (HPx2 and NOX5) that potentiate midgut epithelial nitration in response to Plasmodium infection and regulates expression of two key hemocyte-derived immune effectors (TEP1 and FBN9). Furthermore, the An. gambiae L3-5 strain that has been genetically selected to be refractory (R) to Plasmodium infection exhibits constitutive overexpression of genes from the JNK pathway, as well as midgut and hemocyte effector genes. Silencing experiments confirmed that this cascade mediates, to a large extent, the drastic parasite elimination phenotype characteristic of this mosquito strain. In sum, these studies revealed the JNK pathway as a key regulator of the ability of An. gambiae mosquitoes to limit Plasmodium infection and identified several effector genes mediating these responses. PMID:24039583

  6. The JNK Pathway Is a Key Mediator of Anopheles gambiae Antiplasmodial Immunity

    Science.gov (United States)

    Garver, Lindsey S.; de Almeida Oliveira, Giselle; Barillas-Mury, Carolina

    2013-01-01

    The innate immune system of Anopheles gambiae mosquitoes limits Plasmodium infection through multiple molecular mechanisms. For example, midgut invasion by the parasite triggers an epithelial nitration response that promotes activation of the complement-like system. We found that suppression of the JNK pathway, by silencing either Hep, JNK, Jun or Fos expression, greatly enhanced Plasmodium infection; while overactivating this cascade, by silencing the suppressor Puckered, had the opposite effect. The JNK pathway limits infection via two coordinated responses. It induces the expression of two enzymes (HPx2 and NOX5) that potentiate midgut epithelial nitration in response to Plasmodium infection and regulates expression of two key hemocyte-derived immune effectors (TEP1 and FBN9). Furthermore, the An. gambiae L3–5 strain that has been genetically selected to be refractory (R) to Plasmodium infection exhibits constitutive overexpression of genes from the JNK pathway, as well as midgut and hemocyte effector genes. Silencing experiments confirmed that this cascade mediates, to a large extent, the drastic parasite elimination phenotype characteristic of this mosquito strain. In sum, these studies revealed the JNK pathway as a key regulator of the ability of An. gambiae mosquitoes to limit Plasmodium infection and identified several effector genes mediating these responses. PMID:24039583

  7. Segmental duplication implicated in the genesis of inversion 2Rj of Anopheles gambiae.

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    Mamadou B Coulibaly

    Full Text Available The malaria vector Anopheles gambiae maintains high levels of inversion polymorphism that facilitate its exploitation of diverse ecological settings across tropical Africa. Molecular characterization of inversion breakpoints is a first step toward understanding the processes that generate and maintain inversions. Here we focused on inversion 2Rj because of its association with the assortatively mating Bamako chromosomal form of An. gambiae, whose distinctive breeding sites are rock pools beside the Niger River in Mali and Guinea. Sequence and computational analysis of 2Rj revealed the same 14.6 kb insertion between both breakpoints, which occurred near but not within predicted genes. Each insertion consists of 5.3 kb terminal inverted repeat arms separated by a 4 kb spacer. The insertions lack coding capacity, and are comprised of degraded remnants of repetitive sequences including class I and II transposable elements. Because of their large size and patchwork composition, and as no other instances of these insertions were identified in the An. gambiae genome, they do not appear to be transposable elements. The 14.6 kb modules inserted at both 2Rj breakpoint junctions represent low copy repeats (LCRs, also called segmental duplications that are strongly implicated in the recent (approximately 0.4N(e generations origin of 2Rj. The LCRs contribute to further genome instability, as demonstrated by an imprecise excision event at the proximal breakpoint of 2Rj in field isolates.

  8. Modelling Anopheles gambiae s.s. Population Dynamics with Temperature- and Age-Dependent Survival

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    Christiansen-Jucht, Céline; Erguler, Kamil; Shek, Chee Yan; Basáñez, María-Gloria; Parham, Paul E.

    2015-01-01

    Climate change and global warming are emerging as important threats to human health, particularly through the potential increase in vector- and water-borne diseases. Environmental variables are known to affect substantially the population dynamics and abundance of the poikilothermic vectors of disease, but the exact extent of this sensitivity is not well established. Focusing on malaria and its main vector in Africa, Anopheles gambiae sensu stricto, we present a set of novel mathematical models of climate-driven mosquito population dynamics motivated by experimental data suggesting that in An. gambiae, mortality is temperature and age dependent. We compared the performance of these models to that of a “standard” model ignoring age dependence. We used a longitudinal dataset of vector abundance over 36 months in sub-Saharan Africa for comparison between models that incorporate age dependence and one that does not, and observe that age-dependent models consistently fitted the data better than the reference model. This highlights that including age dependence in the vector component of mosquito-borne disease models may be important to predict more reliably disease transmission dynamics. Further data and studies are needed to enable improved fitting, leading to more accurate and informative model predictions for the An. gambiae malaria vector as well as for other disease vectors. PMID:26030468

  9. Low linkage disequilibrium in wild Anopheles gambiae s.l. populations

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    Harris Caroline

    2010-09-01

    Full Text Available Abstract Background In the malaria vector Anopheles gambiae, understanding diversity in natural populations and genetic components of important phenotypes such as resistance to malaria infection is crucial for developing new malaria transmission blocking strategies. The design and interpretation of many studies here depends critically on Linkage disequilibrium (LD. For example in association studies, LD determines the density of Single Nucleotide Polymorphisms (SNPs to be genotyped to represent the majority of the genomic information. Here, we aim to determine LD in wild An. gambiae s.l. populations in 4 genes potentially involved in mosquito immune responses against pathogens (Gambicin, NOS, REL2 and FBN9 using previously published and newly generated sequences. Results The level of LD between SNP pairs in cloned sequences of each gene was determined for 7 species (or incipient species of the An. gambiae complex. In all tested genes and species, LD between SNPs was low: even at short distances (2 2 ranged from 0.073 to 0.766. In most genes and species LD decayed very rapidly with increasing inter-marker distance. Conclusions These results are of great interest for the development of large scale polymorphism studies, as LD generally falls below any useful limit. It indicates that very fine scale SNP detection will be required to give an overall view of genome-wide polymorphism. Perhaps a more feasible approach to genome wide association studies is to use targeted approaches using candidate gene selection to detect association to phenotypes of interest.

  10. A splice variant of PGRP-LC required for expression of antimicrobial peptides in Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    HUI LIN; LINGMIN ZHANG; CORALIA LUNA; NGO T.HOA; LIANGBIAO ZHENG

    2007-01-01

    Members of the peptidoglycan recognition protein (PGRP) family play essential roles in different manifestations of immune responses in insects. PGRP-LC, one of seven members of this family in the malaria vector Anopheles gambiae produced several spliced variants. Here we show that PGRP-LC, and not other members of the PGRP family nor the six members of the Gram-negative binding protein families, is required for the expression of antimicrobial peptide genes (such as CEC1 and GAM1) under the control of the Imd-Rel2 pathway in an A. gambiae cell line, 4a3A. PGRP-LC produces many splice variants that can be classified into three sub-groups (LC1, LC2 and LC3), based on the carboxyl terminal sequences. RNA interference against one LC1 sub-group resulted in dramatic reduction of CEC1 and GAM1. Over-expression of LC1 a and to a lesser extent LC3a (a member of the LC1 and LC3 sub-group, respectively) in the 4a3A cell line enhances the expression of CEC1 and GAM1. These results demonstrate that the LC1-subgroup splice variants are essential for the expression of CEC1 and GAM1 in A. gambiae cell line.

  11. Multiple insecticide resistance mechanisms in Anopheles gambiae s.l. populations from Cameroon, Central Africa

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    Nwane Philippe

    2013-02-01

    Full Text Available Abstract Background Increasing incidence of DDT and pyrethroid resistance in Anopheles mosquitoes is seen as a limiting factor for malaria vector control. The current study aimed at an in-depth characterization of An. gambiae s.l. resistance to insecticides in Cameroon, in order to guide malaria vector control interventions. Methods Anopheles gambiae s.l. mosquitoes were collected as larvae and pupae from six localities spread throughout the four main biogeographical domains of Cameroon and reared to adults in insectaries. Standard WHO insecticide susceptibility tests were carried out with 4% DDT, 0.75% permethrin and 0.05% deltamethrin. Mortality rates and knockdown times (kdt50 and kdt95 were determined and the effect of pre-exposure to the synergists DEF, DEM and PBO was assessed. Tested mosquitoes were identified to species and molecular forms (M or S using PCR-RFLP. The hot ligation method was used to depict kdr mutations and biochemical assays were conducted to assess detoxifying enzyme activities. Results The An. arabiensis population from Pitoa was fully susceptible to DDT and permethrin (mortality rates > 98% and showed reduced susceptibility to deltamethrin. Resistance to DDT was widespread in An. gambiae s.s. populations and heterogeneous levels of susceptibility to permethrin and deltamethrin were observed. In many cases, prior exposure to synergists partially restored insecticide knockdown effect and increased mortality rates, suggesting a role of detoxifying enzymes in increasing mosquito survival upon challenge by pyrethroids and, to a lower extent DDT. The distribution of kdr alleles suggested a major role of kdr-based resistance in the S form of An. gambiae. In biochemical tests, all but one mosquito population overexpressed P450 activity, whereas baseline GST activity was low and similar in all field mosquito populations and in the control. Conclusion In Cameroon, multiple resistance mechanisms segregate in the S form of An

  12. Spatial and sex-specific dissection of the Anopheles gambiae midgut transcriptome

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    Mahairaki Vassiliki

    2007-01-01

    Full Text Available Abstract Background The midgut of hematophagous insects, such as disease transmitting mosquitoes, carries out a variety of essential functions that mostly relate to blood feeding. The midgut of the female malaria vector mosquito Anopheles gambiae is a major site of interactions between the parasite and the vector. Distinct compartments and cell types of the midgut tissue carry out specific functions and vector borne pathogens interact and infect different parts of the midgut. Results A microarray based global gene expression approach was used to compare transcript abundance in the four major female midgut compartments (cardia, anterior, anterior part of posterior and posterior part of posterior midgut and between the male and female Anopheles gambiae midgut. Major differences between the female and male midgut gene expression relate to digestive processes and immunity. Each compartment has a distinct gene function profile with the posterior midgut expressing digestive enzyme genes and the cardia and anterior midgut expressing high levels of antimicrobial peptide and other immune gene transcripts. Interestingly, the cardia expressed several known anti-Plasmodium factors. A parallel peptidomic analysis of the cardia identified known mosquito antimicrobial peptides as well as several putative short secreted peptides that are likely to represent novel antimicrobial factors. Conclusion The A. gambiae sex specific midgut and female midgut compartment specific transcriptomes correlates with their known functions. The significantly greater functional diversity of the female midgut relate to hematophagy that is associated with digestion and nutrition uptake as well as exposes it to a variety of pathogens, and promotes growth of its endogenous microbial flora. The strikingly high proportion of immunity related factors in the cardia tissue most likely serves the function to increase sterility of ingested sugar and blood. A detailed characterization of the

  13. Genome-wide transcriptional analysis of genes associated with acute desiccation stress in Anopheles gambiae.

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    Mei-Hui Wang

    Full Text Available Malaria transmission in sub-Saharan Africa varies seasonally in intensity. Outbreaks of malaria occur after the beginning of the rainy season, whereas, during the dry season, reports of the disease are less frequent. Anopheles gambiae mosquitoes, the main malaria vector, are observed all year long but their densities are low during the dry season that generally lasts several months. Aestivation, seasonal migration, and local adaptation have been suggested as mechanisms that enable mosquito populations to persist through the dry season. Studies of chromosomal inversions have shown that inversions 2La, 2Rb, 2Rc, 2Rd, and 2Ru are associated with various physiological changes that confer aridity resistance. However, little is known about how phenotypic plasticity responds to seasonally dry conditions. This study examined the effects of desiccation stress on transcriptional regulation in An. gambiae. We exposed female An. gambiae G3 mosquitoes to acute desiccation and conducted a genome-wide analysis of their transcriptomes using the Affymetrix Plasmodium/Anopheles Genome Array. The transcription of 248 genes (1.7% of all transcripts was significantly affected in all experimental conditions, including 96 with increased expression and 152 with decreased expression. In general, the data indicate a reduction in the metabolic rate of mosquitoes exposed to desiccation. Transcripts accumulated at higher levels during desiccation are associated with oxygen radical detoxification, DNA repair and stress responses. The proportion of transcripts within 2La and 2Rs (2Rb, 2Rc, 2Rd, and 2Ru (67/248, or 27% is similar to the percentage of transcripts located within these inversions (31%. These data may be useful in efforts to elucidate the role of chromosomal inversions in aridity tolerance. The scope of application of the anopheline genome demonstrates that examining transcriptional activity in relation to genotypic adaptations greatly expands the number of

  14. Functional characterization of the NF-κB transcription factor gene REL2 from Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    NGO T. HOA; LIANGBIAO ZHENG

    2007-01-01

    The REL2 gene plays an important role in innate immunity against both Gram (+) and Gram (-) bacteria and malaria parasites in Anopheles gambiae, the main vector of malaria in Africa. Through alternative splicing, REL2 produces two protein products, REL2F (with a Rel-homology domain as well as an inhibitory ankyrin repeat region) and REL2S (without the ankyrin repeats). In the immune-competent cell line Sua1B from An. Gambiae, REL2 has been shown to be a key regulator for cecropin A (or CEC1). The high level expression of CEC1 in Sua1B was postulated to be the result of constitutive activation of REL2F. Here we showed that REL2F is indeed processed, albeit at a low level, in the Sua1B cell line. The primary cleavage requires residue 678 (an aspartic acid). Proteolytic cleavage of REL2F can be enhanced by challenge with bacteria Escherichia coli and Bacillus subtilis, but not with fungus Beauveria bassiana. The inducible cleavage can be substantially reduced by RNA interference against PGRP-LC and CASPL1. Over-expression of REL2S or a constitutively active form of REL2F (REL2F380C or REL2F678) in An. Gambiae cell line can further increase expression of CEC1 and other antimicrobial peptide genes. Over-expression of these constitutive active proteins in an immune na?ve cell line, MSQ43, from Anopheles stephensi, results in even more dramatic increased expression of antimicrobial peptides.

  15. Remote sensing and environment in the study of the malaria vector Anopheles gambiae in Mali

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    Rian, Sigrid Katrine Eivindsdatter

    The malaria mosquito Anopheles gambiae is the most important vector for the most devastating form of human malaria, the parasite Plasmodium falciparum. In-depth knowledge of the vector's history and environmental preferences is essential in the pursuit of new malaria mitigation strategies. Research was conducted in Mali across a range of habitats occupied by the vector, focusing on three identified chromosomal forms in the mosquito complex. The development of a 500-m landcover classification map was carried out using MODIS satellite imagery and extensive ground survey. The resulting product has the highest resolution and is the most up-to-date and most extensively ground-surveyed among land-cover maps for the study region. The new landcover classification product is a useful tool in the mapping of the varying ecological preferences of the different An. gambiae chromosomal forms. Climate and vegetation characteristics and their relationship to chromosomal forms were investigated further along a Southwest-Northeast moisture gradient in Mali. This research demonstrates particular ecological preferences of each chromosomal form, and gives a detailed examination of particular vegetation structural and climatological patterns across the study region. A key issue in current research into the population structure of An. gambiae is speciation and evolution in the complex, as an understanding of the mechanisms of change can help in the development of new mitigation strategies. A historical review of the paleoecology, archaeology, and other historical sources intended to shed light on the evolutionary history of the vector is presented. The generally held assumption that the current breed of An. gambiae emerged in the rainforest is called into question and discussed within the framework of paleoenvironment and human expansions in sub-Saharan West Africa.

  16. MicroRNA-regulation of Anopheles gambiae immunity to Plasmodium falciparum infection and midgut microbiota.

    Science.gov (United States)

    Dennison, Nathan J; BenMarzouk-Hidalgo, Omar J; Dimopoulos, George

    2015-03-01

    Invasion of the malaria vector Anopheles gambiae midgut by Plasmodium parasites triggers transcriptional changes of immune genes that mediate the antiparasitic defense. This response is largely regulated by the Toll and Immune deficiency (IMD) pathways. To determine whether A. gambiae microRNAs (miRNAs) are involved in regulating the anti-Plasmodium defense, we showed that suppression of miRNA biogenesis results in increased resistance to Plasmodium falciparum infection. In silico analysis of A. gambiae immune effector genes identified multiple transcripts with miRNA binding sites. A comparative miRNA microarray abundance analysis of P. falciparum infected and naïve mosquito midgut tissues showed elevated abundance of miRNAs aga-miR-989 and aga-miR-305 in infected midguts. Antagomir inhibition of aga-miR-305 increased resistance to P. falciparum infection and suppressed the midgut microbiota. Conversely, treatment of mosquitoes with an artificial aga-miR-305 mimic increased susceptibility to P. falciparum infection and resulted in expansion of midgut microbiota, suggesting that aga-miR-305 acts as a P. falciparum and gut microbiota agonist by negatively regulating the mosquito immune response. In silico prediction of aga-miR-305 target genes identified several anti-Plasmodium effectors. Our study shows that A. gambiae aga-miR-305 regulates the anti-Plasmodium response and midgut microbiota, likely through post-transcriptional modification of immune effector genes.

  17. Chromosome inversions, genomic differentiation and speciation in the African malaria mosquito Anopheles gambiae.

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    Yoosook Lee

    Full Text Available The African malaria vector, Anopheles gambiae, is characterized by multiple polymorphic chromosomal inversions and has become widely studied as a system for exploring models of speciation. Near complete reproductive isolation between different inversion types, known as chromosomal forms, has led to the suggestion that A. gambiae is in early stages of speciation, with divergence evolving in the face of considerable gene flow. We compared the standard chromosomal arrangement (Savanna form with genomes homozygous for j, b, c, and u inversions (Bamako form in order to identify regions of genomic divergence with respect to inversion polymorphism. We found levels of divergence between the two sub-taxa within some of these inversions (2Rj and 2Rb, but at a level lower than expected and confined near the inversion breakpoints, consistent with a gene flux model. Unexpectedly, we found that the majority of diverged regions were located on the X chromosome, which contained half of all significantly diverged regions, with much of this divergence located within exons. This is surprising given that the Bamako and Savanna chromosomal forms are both within the S molecular form that is defined by a locus near centromere of X chromosome. Two X-linked genes (a heat shock protein and P450 encoding genes involved in reproductive isolation between the M and S molecular forms of A. gambiae were also significantly diverged between the two chromosomal forms. These results suggest that genes mediating reproductive isolation are likely located on the X chromosome, as is thought to be the case for the M and S molecular forms. We conclude that genes located on the sex chromosome may be the major force driving speciation between these chromosomal forms of A. gambiae.

  18. The interplay between tubulins and P450 cytochromes during Plasmodium berghei invasion of Anopheles gambiae midgut.

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    Rute C Félix

    Full Text Available BACKGROUND: Plasmodium infection increases the oxidative stress inside the mosquito, leading to a significant alteration on transcription of Anopheles gambiae detoxification genes. Among these detoxification genes several P450 cytochromes and tubulins were differently expressed, suggesting their involvement in the mosquito's response to parasite invasion. P450 cytochromes are usually involved in the metabolism and detoxification of several compounds, but are also regulated by several pathogens, including malaria parasite. Tubulins are extremely important as components of the cytoskeleton, which rearrangement functions as a response to malaria parasite invasion. METHODOLOGY/PRINCIPAL FINDINGS: Gene silencing methods were used to uncover the effects of cytochrome P450 reductase, tubulinA and tubulinB silencing on the A. gambiae response to Plasmodium berghei invasion. The role of tubulins in counter infection processes was also investigated by inhibiting their effect. Colchicine, vinblastine and paclitaxel, three different tubulin inhibitors were injected into A. gambiae mosquitoes. Twenty-four hours post injection these mosquitoes were infected with P. berghei through a blood meal from infected CD1 mice. Cytochrome P450 gene expression was measured using RT-qPCR to detect differences in cytochrome expression between silenced, inhibited and control mosquitoes. Results showed that cytochrome P450 reductase silencing, as well as tubulin (A and B silencing and inhibition affected the efficiency of Plasmodium infection. Silencing and inhibition also affected the expression levels of cytochromes P450. CONCLUSIONS: Our results suggest the existence of a relationship between tubulins and P450 cytochromes during A. gambiae immune response to P. berghei invasion. One of the P450 cytochromes in this study, CYP6Z2, stands out as the potential link in this association. Further work is needed to fully understand the role of tubulin genes in the response to

  19. Influence of Land-use on the Fitness of Anopheles gambiae, the Principal Vector of Malaria in Nigeria

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    Israel Kayode Olayemi

    2009-02-01

    Full Text Available Background: Urbanization often results in profound environmental alterations that may promote the transmission of malaria. Though, land-use practices in urban areas have been linked with proliferations of suitable larval breeding habitats of malaria vectors, no attempt has been made to systematically investigate the influence of land-use practices on malaria transmission in Nigeria. Objectives: To elucidate the influence of land-use practices on larval development and adult body size of Anopheles gambiae (Diptera: Culicidae mosquitoes in Minna, Nigeria. Materials and Methods: Newly-hatched larvae of An. gmbiae mosquitoes were reared in semi-natural habitats stationed in five different sites, each representing the major land-use types in the area. The larvae were monitored daily for Duration of Immature Development (DID and Immature Survival Rate (ISR; while Wing Length (WL was used as an index of adult body size. Results: DID, ISR and WL varied significantly (P < 0.05 among the land-use categories; with lager numbers of bigger mosquitoes produced at a faster rate in the artificial than natural land-use sites. Water temperature for larval development was best in the Refuse Dump (RD site (mean = 28.11 ± 2.50oC and consequently the shortest DID (mean = 9.70 ± 0.74 days, as well as, the largest mosquitoes (mean WL = 3.10 ± 0.90 mm, were recorded in this land-use category. However, while ISR was highest (mean = 96.30 ± 2.78% in Farm Land (FL, the mosquitoes that emerged from this site were the smallest (mean WL = 1.96 ± 0.51mm. The Natural Vegetation (NV land-use category was the least productive, as the larvae took the longest time (13.29 ± 1.69 days to develop, and survived least (42.94 ± 7.50% in this site. Conclusion: The land-use practices in Minna enhanced the fitness of An. gambiae, and may increase the vectorial capacity of the species for malaria transmission in the area. Targeted larviciding interventions will greatly contribute to

  20. Development of a Gravid Trap for Collecting Live Malaria Vectors Anopheles gambiae s.l.

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    Dugassa, Sisay; Lindh, Jenny M.; Oyieke, Florence; Mukabana, Wolfgang R.; Lindsay, Steven W.; Fillinger, Ulrike

    2013-01-01

    Background Effective malaria vector control targeting indoor host-seeking mosquitoes has resulted in fewer vectors entering houses in many areas of sub-Saharan Africa, with the proportion of vectors outdoors becoming more important in the transmission of this disease. This study aimed to develop a gravid trap for the outdoor collection of the malaria vector Anopheles gambiae s.l. based on evaluation and modification of commercially available gravid traps. Methods Experiments were implemented in an 80 m2 semi-field system where 200 gravid Anopheles gambiae s.s. were released nightly. The efficacy of the Box, CDC and Frommer updraft gravid traps was compared. The Box gravid trap was tested to determine if the presence of the trap over water and the trap’s sound affected catch size. Mosquitoes approaching the treatment were evaluated using electrocuting nets or detergents added to the water in the trap. Based on the results, a new gravid trap (OviART trap) that provided an open, unobstructed oviposition site was developed and evaluated. Results Box and CDC gravid traps collected similar numbers (relative rate (RR) 0.8, 95% confidence interval (CI) 0.6–1.2; p = 0.284), whereas the Frommer trap caught 70% fewer mosquitoes (RR 0.3, 95% CI 0.2–0.5; p < 0.001). The number of mosquitoes approaching the Box trap was significantly reduced when the trap was positioned over a water-filled basin compared to an open pond (RR 0.7 95% CI 0.6–0.7; p < 0.001). This effect was not due to the sound of the trap. Catch size increased by 60% (RR 1.6, 1.2–2.2; p = 0.001) with the new OviART trap. Conclusion Gravid An. Gambiae s.s. females were visually deterred by the presence of the trapping device directly over the oviposition medium. Based on these investigations, an effective gravid trap was developed that provides open landing space for egg-laying Anopheles. PMID:23861952

  1. Development of a gravid trap for collecting live malaria vectors Anopheles gambiae s.l.

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    Sisay Dugassa

    Full Text Available BACKGROUND: Effective malaria vector control targeting indoor host-seeking mosquitoes has resulted in fewer vectors entering houses in many areas of sub-Saharan Africa, with the proportion of vectors outdoors becoming more important in the transmission of this disease. This study aimed to develop a gravid trap for the outdoor collection of the malaria vector Anopheles gambiae s.l. based on evaluation and modification of commercially available gravid traps. METHODS: Experiments were implemented in an 80 m(2 semi-field system where 200 gravid Anopheles gambiae s.s. were released nightly. The efficacy of the Box, CDC and Frommer updraft gravid traps was compared. The Box gravid trap was tested to determine if the presence of the trap over water and the trap's sound affected catch size. Mosquitoes approaching the treatment were evaluated using electrocuting nets or detergents added to the water in the trap. Based on the results, a new gravid trap (OviART trap that provided an open, unobstructed oviposition site was developed and evaluated. RESULTS: Box and CDC gravid traps collected similar numbers (relative rate (RR 0.8, 95% confidence interval (CI 0.6-1.2; p = 0.284, whereas the Frommer trap caught 70% fewer mosquitoes (RR 0.3, 95% CI 0.2-0.5; p < 0.001. The number of mosquitoes approaching the Box trap was significantly reduced when the trap was positioned over a water-filled basin compared to an open pond (RR 0.7 95% CI 0.6-0.7; p < 0.001. This effect was not due to the sound of the trap. Catch size increased by 60% (RR 1.6, 1.2-2.2; p = 0.001 with the new OviART trap. CONCLUSION: Gravid An. Gambiae s.s. females were visually deterred by the presence of the trapping device directly over the oviposition medium. Based on these investigations, an effective gravid trap was developed that provides open landing space for egg-laying Anopheles.

  2. Toxicity of essential oil from Indian borage on the larvae of the African malaria vector mosquito, Anopheles gambiae

    OpenAIRE

    Kweka Eliningaya J; Senthilkumar Annadurai; Venkatesalu Venugopalan

    2012-01-01

    Abstract Background Essential oils are currently studied for the control of different disease vectors, because of their efficacy on targeted organisms. In the present investigation, the larvicidal potential of essential oil extracted from Indian borage (Plectranthus amboinicus) was studied against the African anthropophagic malaria vector mosquito, Anopheles gambiae. The larvae of An. gambiae s.s laboratory colony and An. gambiae s.l of wild populations were assayed and the larval mortality w...

  3. Chemical composition and insecticidal activity of plant essential oils from Benin against Anopheles gambiae (Giles)

    Science.gov (United States)

    2013-01-01

    Background Insecticide resistance in sub-Saharan Africa and especially in Benin is a major public health issue hindering the control of the malaria vectors. Each Anopheles species has developed a resistance to one or several classes of the insecticides currently in use in the field. Therefore, it is urgent to find alternative compounds to conquer the vector. In this study, the efficacies of essential oils of nine plant species, which are traditionally used to avoid mosquito bites in Benin, were investigated. Methods Essential oils of nine plant species were extracted by hydrodistillation, and their chemical compositions were identified by GC-MS. These oils were tested on susceptible “kisumu” and resistant “ladji-Cotonou” strains of Anopheles gambiae, following WHO test procedures for insecticide resistance monitoring in malaria vector mosquitoes. Results Different chemical compositions were obtained from the essential oils of the plant species. The major constituents identified were as follows: neral and geranial for Cymbopogon citratus, Z-carveol, E-p-mentha-1(7),8-dien-2-ol and E-p-mentha-2,8-dienol for Cymbopogon giganteus, piperitone for Cymbopogon schoenanthus, citronellal and citronellol for Eucalyptus citriodora, p-cymene, caryophyllene oxide and spathulenol for Eucalyptus tereticornis, 3-tetradecanone for Cochlospermum tinctorium and Cochlospermum planchonii, methyl salicylate for Securidaca longepedunculata and ascaridole for Chenopodium ambrosioides. The diagnostic dose was 0.77% for C. citratus, 2.80% for E. tereticornis, 3.37% for E. citriodora, 4.26% for C. ambrosioides, 5.48% for C. schoenanthus and 7.36% for C. giganteus. The highest diagnostic doses were obtained with S. longepedunculata (9.84%), C. tinctorium (11.56%) and C. planchonii (15.22%), compared to permethrin 0.75%. A. gambiae cotonou, which is resistant to pyrethroids, showed significant tolerance to essential oils from C. tinctorium and S. longepedunculata as expected but was

  4. Reactive oxygen species detoxification by catalase is a major determinant of fecundity in the mosquito Anopheles gambiae

    OpenAIRE

    DeJong, Randall J.; Miller, Lisa M; Molina-Cruz, Alvaro; Gupta, Lalita; Kumar, Sanjeev; Barillas-Mury, Carolina

    2007-01-01

    The mosquito Anopheles gambiae is a primary vector of Plasmodium parasites in Africa. The effect of aging on reproductive output in A. gambiae females from three strains that differ in their ability to melanize Plasmodium and in their systemic levels of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), was analyzed. The number of eggs oviposited after the first blood meal decreases with age in all strains; however, this decline was much more pronounced in the G3 (unselected) and R (r...

  5. Larvicidal activity of extracts from three Plumbago spp against Anopheles gambiae

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    Barasa M Maniafu

    2009-09-01

    Full Text Available Three Plumbago spp have been tested for mosquito larvicidal activity. The crude extracts exhibiting the highest larvicidal activity against Anopheles gambiae were hexane (LC50 = 6.4 μg/mL and chloroform (LC50 = 6.7 μg/mL extracts from Plumbago zeylanica Linn, chloroform (LC50 = 6.7 ug/mL extract from Plumbago stenophylla Bull and ethyl acetate (LC50 = 4.1 μg/mL extract from Plumbago dawei Rolfe. These LC50 values were within 95% confidence limits. 5-hydroxy-2-methyl-1,4-naphthoquinone (plumbagin 1 (LC50 = 1.9 μg/mL and β-sitosterol 2 were characterised from ethyl acetate extract of root bark of P. dawei, a native medicinal plant growing in Kenya, based on spectral analysis and comparisons with data in literature.

  6. A mosquito lipoxin/lipocalin complex mediates innate immune priming in Anopheles gambiae.

    Science.gov (United States)

    Ramirez, Jose Luis; de Almeida Oliveira, Giselle; Calvo, Eric; Dalli, Jesmond; Colas, Romain A; Serhan, Charles N; Ribeiro, Jose M; Barillas-Mury, Carolina

    2015-01-01

    Exposure of Anopheles gambiae mosquitoes to Plasmodium infection enhances the ability of their immune system to respond to subsequent infections. However, the molecular mechanism that allows the insect innate immune system to 'remember' a previous encounter with a pathogen has not been established. Challenged mosquitoes constitutively release a soluble haemocyte differentiation factor into their haemolymph that, when transferred into Naive mosquitoes, also induces priming. Here we show that this factor consists of a Lipoxin/Lipocalin complex. We demonstrate that innate immune priming in mosquitoes involves a persistent increase in expression of Evokin (a lipid carrier of the lipocalin family), and in their ability to convert arachidonic acid to lipoxins, predominantly Lipoxin A4. Plasmodium ookinete midgut invasion triggers immune priming by inducing the release of a mosquito lipoxin/lipocalin complex. PMID:26100162

  7. Genome expression analysis of Anopheles gambiae: responses to injury, bacterial challenge, and malaria infection.

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    Dimopoulos, George; Christophides, George K; Meister, Stephan; Schultz, Jörg; White, Kevin P; Barillas-Mury, Carolina; Kafatos, Fotis C

    2002-06-25

    The complex gene expression responses of Anopheles gambiae to microbial and malaria challenges, injury, and oxidative stress (in the mosquito and/or a cultured cell line) were surveyed by using cDNA microarrays constructed from an EST-clone collection. The expression profiles were broadly subdivided into induced and down-regulated gene clusters. Gram+ and Gram- bacteria and microbial elicitors up-regulated a diverse set of genes, many belonging to the immunity class, and the response to malaria partially overlapped with this response. Oxidative stress activated a distinctive set of genes, mainly implicated in oxidoreductive processes. Injury up- and down-regulated gene clusters also were distinctive, prominently implicating glycolysis-related genes and citric acid cycle/oxidative phosphorylation/redox-mitochondrial functions, respectively. Cross-comparison of in vivo and in vitro responses indicated the existence of tightly coregulated gene groups that may correspond to gene pathways. PMID:12077297

  8. A mosquito lipoxin/lipocalin complex mediates innate immune priming in Anopheles gambiae

    Science.gov (United States)

    Ramirez, Jose Luis; de Almeida Oliveira, Giselle; Calvo, Eric; Dalli, Jesmond; Colas, Romain A.; Serhan, Charles N.; Ribeiro, Jose M.; Barillas-Mury, Carolina

    2015-01-01

    Exposure of Anopheles gambiae mosquitoes to Plasmodium infection enhances the ability of their immune system to respond to subsequent infections. However, the molecular mechanism that allows the insect innate immune system to ‘remember' a previous encounter with a pathogen has not been established. Challenged mosquitoes constitutively release a soluble haemocyte differentiation factor into their haemolymph that, when transferred into Naive mosquitoes, also induces priming. Here we show that this factor consists of a Lipoxin/Lipocalin complex. We demonstrate that innate immune priming in mosquitoes involves a persistent increase in expression of Evokin (a lipid carrier of the lipocalin family), and in their ability to convert arachidonic acid to lipoxins, predominantly Lipoxin A4. Plasmodium ookinete midgut invasion triggers immune priming by inducing the release of a mosquito lipoxin/lipocalin complex. PMID:26100162

  9. West African Anopheles gambiae mosquitoes harbor a taxonomically diverse virome including new insect-specific flaviviruses, mononegaviruses, and totiviruses.

    Science.gov (United States)

    Fauver, Joseph R; Grubaugh, Nathan D; Krajacich, Benjamin J; Weger-Lucarelli, James; Lakin, Steven M; Fakoli, Lawrence S; Bolay, Fatorma K; Diclaro, Joseph W; Dabiré, Kounbobr Roch; Foy, Brian D; Brackney, Doug E; Ebel, Gregory D; Stenglein, Mark D

    2016-11-01

    Anopheles gambiae are a major vector of malaria in sub-Saharan Africa. Viruses that naturally infect these mosquitoes may impact their physiology and ability to transmit pathogens. We therefore used metagenomics sequencing to search for viruses in adult Anopheles mosquitoes collected from Liberia, Senegal, and Burkina Faso. We identified a number of virus and virus-like sequences from mosquito midgut contents, including 14 coding-complete genome segments and 26 partial sequences. The coding-complete sequences define new viruses in the order Mononegavirales, and the families Flaviviridae, and Totiviridae. The identification of a flavivirus infecting Anopheles mosquitoes broadens our understanding of the evolution and host range of this virus family. This study increases our understanding of virus diversity in general, begins to define the virome of a medically important vector in its natural setting, and lays groundwork for future studies examining the potential impact of these viruses on anopheles biology and disease transmission. PMID:27639161

  10. Comprehensive genetic dissection of the hemocyte immune response in the malaria mosquito Anopheles gambiae.

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    Fabrizio Lombardo

    2013-01-01

    Full Text Available Reverse genetics in the mosquito Anopheles gambiae by RNAi mediated gene silencing has led in recent years to an advanced understanding of the mosquito immune response against infections with bacteria and malaria parasites. We developed RNAi screens in An. gambiae hemocyte-like cells using a library of double-stranded RNAs targeting 109 genes expressed highly or specifically in mosquito hemocytes to identify novel regulators of the hemocyte immune response. Assays included phagocytosis of bacterial bioparticles, expression of the antimicrobial peptide CEC1, and basal and induced expression of the mosquito complement factor LRIM1. A cell viability screen was also carried out to assess dsRNA cytotoxicity and to identify genes involved in cell growth and survival. Our results identify 22 novel immune regulators, including proteins putatively involved in phagosome assembly and maturation (Ca²⁺ channel, v-ATPase and cyclin-dependent protein kinase, pattern recognition (fibrinogen-domain lectins and Nimrod, immune modulation (peptidase and serine protease homolog, immune signaling (Eiger and LPS-induced factor, cell adhesion and communication (Laminin B1 and Ninjurin and immune homeostasis (Lipophorin receptor. The development of robust functional cell-based assays paves the way for genome-wide functional screens to study the mosquito immune response to infections with human pathogens.

  11. Functional characterization of PGRP-LC1 of Anopheles gambiae through deletion and RNA interference

    Institute of Scientific and Technical Information of China (English)

    Yang Chen; Erjun Ling; Zhihui Weng

    2009-01-01

    Peptidoglycan recognition proteins (PGRP) play an important role in innate immunity in insects through the activation of the Imd pathway, which has been shown to be required in the antibacterial response in insects and in the limitation of the number of Plasmodium berghei oocysts developing in mosquito midgut. The LC1gene of the PRGP family in Anopheles gambiae produces many products through alternative splicing. In this work, we demonstrate that PGRP-LC1a alone is sufficient to activate the Imd pathway in the A. gambiae L3-5 cell line through a combination of terminal or internal deletions, and RNA interference against endogenous PGRP-LC products. In the absence of endogenous PGRP-LC proteins, the integrity of the cytoplasmic domain is necessary for LC1 a function, while that of the extracellular domain is not. Moreover, the shorter the extracellular domain, the higher the activity for LC1a. However, the removal of either the cytoplasmic or the extracellular PGRP-binding domain has little impact on the activity of LC1a in the presence of endogenous PGRP-LC proteins.

  12. The STAT pathway mediates late phase immunity against Plasmodium in the mosquito Anopheles gambiae

    Science.gov (United States)

    Gupta, Lalita; Molina-Cruz, Alvaro; Kumar, Sanjeev; Rodrigues, Janneth; Dixit, Rajnikant; Zamora, Rodolfo E.; Barillas-Mury, Carolina

    2009-01-01

    The STAT family of transcription factors activate expression of immune system genes in vertebrates. The ancestral STAT gene (AgSTAT-A) appears to have duplicated in the mosquito Anopheles gambiae, giving rise to a second intronless STAT gene (AgSTAT-B), which we show regulates AgSTAT-A expression in adult females. AgSTAT-A participates in the transcriptional activation of nitric oxide synthase (NOS) in response to bacterial and plasmodial infection. Activation of this pathway, however, is not essential for mosquitoes to survive a bacterial challenge. AgSTAT-A silencing reduces the number of early Plasmodium oocysts in the midgut, but nevertheless enhances the overall infection by increasing oocyst survival. Silencing of SOCS, a STAT suppressor, has the opposite effect, reducing Plasmodium infection by increasing NOS expression. Chemical inhibition of mosquito NOS activity after oocyte formation increases oocyte survival. Thus, the AgSTAT-A pathway mediates a late phase anti-plasmodial response that reduces oocyst survival in An. gambiae. PMID:19454353

  13. The STAT pathway mediates late-phase immunity against Plasmodium in the mosquito Anopheles gambiae.

    Science.gov (United States)

    Gupta, Lalita; Molina-Cruz, Alvaro; Kumar, Sanjeev; Rodrigues, Janneth; Dixit, Rajnikant; Zamora, Rodolfo E; Barillas-Mury, Carolina

    2009-05-01

    The STAT family of transcription factors activates expression of immune system genes in vertebrates. The ancestral STAT gene (AgSTAT-A) appears to have duplicated in the mosquito Anopheles gambiae, giving rise to a second intronless STAT gene (AgSTAT-B), which we show regulates AgSTAT-A expression in adult females. AgSTAT-A participates in the transcriptional activation of nitric oxide synthase (NOS) in response to bacterial and plasmodial infection. Activation of this pathway, however, is not essential for mosquitoes to survive a bacterial challenge. AgSTAT-A silencing reduces the number of early Plasmodium oocysts in the midgut, but nevertheless enhances the overall infection by increasing oocyst survival. Silencing of SOCS, a STAT suppressor, has the opposite effect, reducing Plasmodium infection by increasing NOS expression. Chemical inhibition of mosquito NOS activity after oocyte formation increases oocyte survival. Thus, the AgSTAT-A pathway mediates a late-phase antiplasmodial response that reduces oocyst survival in A. gambiae. PMID:19454353

  14. Insights into the epigenomic landscape of the human malaria vector Anopheles gambiae

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    Elena eGómez-Díaz

    2014-08-01

    Full Text Available The epigenome of the human malaria vector Anopheles gambiae was characterized in midgut cells by mapping the distribution and levels of two post-translational histone modifications, H3K27ac and H3K27me3. These histone profiles were then correlated with levels of gene expression obtained by RNA-seq. Analysis of the transcriptome of A. gambiae midguts and salivary glands led to the discovery of 13,898 new transcripts not present in the most recent genome assembly. A subset of these transcripts is differentially expressed between midgut and salivary glands. The enrichment profiles of H3K27ac and H3K27me3 are mutually exclusive and associate with high and low levels of transcription, respectively. This distribution agrees with previous findings in Drosophila showing association of these two histone modifications with either active or inactive transcriptional states, including Polycomb-associated domains in silenced genes. This study provides a mosquito epigenomics platform for future comparative studies in other mosquito species, opening future investigations into the role of epigenetic processes in vector-borne systems of medical and economic importance.

  15. Reactive oxygen species modulate Anopheles gambiae immunity against bacteria and Plasmodium.

    Science.gov (United States)

    Molina-Cruz, Alvaro; DeJong, Randall J; Charles, Bradley; Gupta, Lalita; Kumar, Sanjeev; Jaramillo-Gutierrez, Giovanna; Barillas-Mury, Carolina

    2008-02-01

    The involvement of reactive oxygen species (ROS) in mosquito immunity against bacteria and Plasmodium was investigated in the malaria vector Anopheles gambiae. Strains of An. gambiae with higher systemic levels of ROS survive a bacterial challenge better, whereas reduction of ROS by dietary administration of antioxidants significantly decreases survival, indicating that ROS are required to mount effective antibacterial responses. Expression of several ROS detoxification enzymes increases in the midgut and fat body after a blood meal. Furthermore, expression of several of these enzymes increases to even higher levels when mosquitoes are fed a Plasmodium berghei-infected meal, indicating that the oxidative stress after a blood meal is exacerbated by Plasmodium infection. Paradoxically, a complete lack of induction of catalase mRNA and lower catalase activity were observed in P. berghei-infected midguts. This suppression of midgut catalase expression is a specific response to ookinete midgut invasion and is expected to lead to higher local levels of hydrogen peroxide. Further reduction of catalase expression by double-stranded RNA-mediated gene silencing promoted parasite clearance by a lytic mechanism and reduced infection significantly. High mosquito mortality is often observed after P. berghei infection. Death appears to result in part from excess production of ROS, as mortality can be decreased by oral administration of uric acid, a strong antioxidant. We conclude that ROS modulate An. gambiae immunity and that the mosquito response to P. berghei involves a local reduction of detoxification of hydrogen peroxide in the midgut that contributes to limit Plasmodium infection through a lytic mechanism. PMID:18065421

  16. Transcription regulation of sex-biased genes during ontogeny in the malaria vector Anopheles gambiae.

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    Kalle Magnusson

    Full Text Available In Anopheles gambiae, sex-regulated genes are responsible for controlling gender dimorphism and are therefore crucial in determining the ability of female mosquitoes to transmit human malaria. The identification and functional characterization of these genes will shed light on the sexual development and maturation of mosquitoes and provide useful targets for genetic control measures aimed at reducing mosquito fertility and/or distorting the sex ratio.We conducted a genome wide transcriptional analysis of sex-regulated genes from early developmental stages through adulthood combined with functional screening of novel gonadal genes. Our results demonstrate that the male-biased genes undergo a major transcription turnover starting from larval stages to adulthood. The male biased genes at the adult stage include a significant high number of unique sequences compared to the rest of the genome. This is in contrast to female-biased genes that are much more conserved and are mainly activated during late developmental stages.The high frequency of unique sequences would indicate that male-biased genes evolve more rapidly than the rest of the genome. This finding is particularly intriguing because A. gambiae is a strictly female monogamous species suggesting that driving forces in addition to sperm competition must account for the rapid evolution of male-biased genes. We have also identified and functionally characterized a number of previously unknown A. gambiae testis- and ovary-specific genes. Two of these genes, zero population growth and a suppressor of defective silencing 3 domain of the histone deacetylase co-repressor complex, were shown to play a key role in gonad development.

  17. Biochemical characterization of chitin synthase activity and inhibition in the African malaria mosquito, Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    Xin Zhang; Kun Yan Zhu

    2013-01-01

    Chitin synthase (CHS) is an important enzyme catalyzing the formation of chitin polymers in all chitin containing organisms and a potential target site for insect pest control.However,our understanding of biochemical properties of insect CHSs has been very limited.We here report enzymatic and inhibitory properties of CHS prepared from the African malaria mosquito,Anopheles gambiae.Our study,which represents the first time to use a nonradioactive method to assay CHS activity in an insect species,determined the optimal conditions for measuring the enzyme activity,including pH,temperature,and concentrations of the substrate uridine diphosphate N-acetyl-D-glucosamine (UDPGlcNAc) and Mg++.The optimal pH was about 6.5-7.0,and the highest activity was detected at temperatures between 37℃ and 44℃.Dithithreitol is required to prevent melanization of the enzyme extract.CHS activity was enhanced at low concentration of GlcNAc,but inhibited at high concentrations.Proteolytic activation of the activity is significant both in the 500×g supernatant and the 40 000×g pellet.Our study revealed only slight in vitro inhibition ofA.gambiae CHS activity by diflubenzuron and nikkomycin Z at the highest concentration (2.5μmol/L) examined.There was no in vitro inhibition by polyoxin D at any concentration examined.Furthermore,we did not observe any in vivo inhibition of CHS activity by any of these chemicals at any concentration examined.Our results suggest that the inhibition of chitin synthesis by these chemicals is not due to direct inhibition of CHS in A.gambiae.

  18. P elements and MITE relatives in the whole genome sequence of Anopheles gambiae

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    Nouaud Danielle

    2006-08-01

    Full Text Available Abstract Background Miniature Inverted-repeat Terminal Elements (MITEs, which are particular class-II transposable elements (TEs, play an important role in genome evolution, because they have very high copy numbers and display recurrent bursts of transposition. The 5' and 3' subterminal regions of a given MITE family often show a high sequence similarity with the corresponding regions of an autonomous Class-II TE family. However, the sustained presence over a prolonged evolutionary time of MITEs and TE master copies able to promote their mobility has been rarely reported within the same genome, and this raises fascinating evolutionary questions. Results We report here the presence of P transposable elements with related MITE families in the Anopheles gambiae genome. Using a TE annotation pipeline we have identified and analyzed all the P sequences in the sequenced A. gambiae PEST strain genome. More than 0.49% of the genome consists of P elements and derivates. P elements can be divided into 9 different subfamilies, separated by more than 30% of nucleotide divergence. Seven of them present full length copies. Ten MITE families are associated with 6 out of the 9 Psubfamilies. Comparing their intra-element nucleotide diversities and their structures allows us to propose the putative dynamics of their emergence. In particular, one MITE family which has a hybrid structure, with ends each of which is related to a different P-subfamily, suggests a new mechanism for their emergence and their mobility. Conclusion This work contributes to a greater understanding of the relationship between full-length class-II TEs and MITEs, in this case P elements and their derivatives in the genome of A. gambiae. Moreover, it provides the most comprehensive catalogue to date of P-like transposons in this genome and provides convincing yet indirect evidence that some of the subfamilies have been recently active.

  19. Indirect evidence that agricultural pesticides select for insecticide resistance in the malaria vector Anopheles gambiae.

    Science.gov (United States)

    Luc, Djogbénou S; Benoit, Assogba; Laurette, Djossou; Michel, Makoutode

    2016-06-01

    We investigated the possible relationship between the agricultural use of insecticides and the emergence of insecticide resistance. Bioassays were conducted using simulated mosquito larval habitats and well known Anopheles gambiae strains. Soil samples were collected from vegetable production areas in Benin, including one site with insecticide use, one site where insecticides had not been used for two months, and a third where insecticides had not been used. Pupation and emergence rates were very low in pyrethroid-susceptible strains when exposed to soil that had been recently exposed to insecticides. Pupation and emergence rates in strains with the kdr mutation alone or both the kdr and Ace-1 mutations were much higher. Overall, strains with the kdr mutation survived at higher rates compared to that without kdr mutation. Although this study is observational, we provide indirect evidence indicating that soils from agricultural areas contain insecticide residues that can play a role in the emergence of insecticide resistance in Anopheles. This aspect should be taken into account to better utilize the insecticide in the context of integrated pest management programs.

  20. Population genetics of Plasmodium resistance genes in Anopheles gambiae: no evidence for strong selection.

    Science.gov (United States)

    Obbard, D J; Linton, Y-M; Jiggins, F M; Yan, G; Little, T J

    2007-08-01

    Anopheles mosquitoes are the primary vectors for malaria in Africa, transmitting the disease to more than 100 million people annually. Recent functional studies have revealed mosquito genes that are crucial for Plasmodium development, but there is presently little understanding of which genes mediate vector competence in the wild, or evolve in response to parasite-mediated selection. Here, we use population genetic approaches to study the strength and mode of natural selection on a suite of mosquito immune system genes, CTL4, CTLMA2, LRIM1, and APL2 (LRRD7), which have been shown to affect Plasmodium development in functional studies. We sampled these genes from two African populations of An. gambiae s.s., along with several closely related species, and conclude that there is no evidence for either strong directional or balancing selection on these genes. We highlight a number of challenges that need to be met in order to apply population genetic tests for selection in Anopheles mosquitoes; in particular the dearth of suitable outgroup species and the potential difficulties that arise when working within a closely-related species complex. PMID:17688548

  1. Effect of discriminative plant-sugar feeding on the survival and fecundity of Anopheles gambiae

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    Jackson Robert R

    2007-08-01

    Full Text Available Abstract Background A previous study showed for Anopheles gambiae s.s. a gradation of feeding preference on common plant species growing in a malaria holoendemic area in western Kenya. The present follow-up study determines whether there is a relationship between the mosquito's preferences and its survival and fecundity. Methods Groups of mosquitoes were separately given ad libitum opportunity to feed on five of the more preferred plant species (Hamelia patens, Parthenium hysterophorus, Ricinus communis, Senna didymobotrya, and Tecoma stans and one of the less preferred species (Lantana camara. The mosquitoes were monitored daily for survival. Sugar solution (glucose 6% and water were used as controls. In addition, the fecundity of mosquitoes on each plant after (i only one blood meal (number of eggs oviposited, and (ii after three consecutive blood meals (proportion of females ovipositing, number of eggs oviposited and hatchability of eggs, was determined. The composition and concentration of sugar in the fed-on parts of each plant species were determined using gas chromatography. Using SAS statistical package, tests for significant difference of the fitness values between mosquitoes exposed to different plant species were conducted. Results and Conclusion Anopheles gambiae that had fed on four of the five more preferred plant species (T. stans, S. didymobotrya, R. communis and H. patens, but not P. hysterophorus lived longer and laid more eggs after one blood meal, when compared with An. gambiae that had fed on the least preferred plant species L. camara. When given three consecutive blood-meals, the percentage of females that oviposited, but not the number of eggs laid, was significantly higher for mosquitoes that had previously fed on the four more preferred plant species. Total sugar concentration in the preferred plant parts was significantly correlated with survival and with the proportion of females that laid eggs. This effect was

  2. A major genetic locus controlling natural Plasmodium falciparum infection is shared by East and West African Anopheles gambiae

    OpenAIRE

    Koella Jacob C; Sharakhov Igor; Xia Ai; Lambrechts Louis; Markianos Kyriacos; Riehle Michelle M; Vernick Kenneth D

    2007-01-01

    Abstract Background Genetic linkage mapping identified a region of chromosome 2L in the Anopheles gambiae genome that exerts major control over natural infection by Plasmodium falciparum. This 2L Plasmodium-resistance interval was mapped in mosquitoes from a natural population in Mali, West Africa, and controls the numbers of P. falciparum oocysts that develop on the vector midgut. An important question is whether genetic variation with respect to Plasmodium-resistance exists across Africa, a...

  3. An Epithelial Serine Protease, AgESP, Is Required for Plasmodium Invasion in the Mosquito Anopheles gambiae

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    Rodrigues, Janneth; Oliveira, Giselle A.; Kotsyfakis, Michalis; Dixit, Rajnikant; Molina-Cruz, Alvaro; Jochim, Ryan; Barillas-Mury, Carolina

    2012-01-01

    Background Plasmodium parasites need to cross the midgut and salivary gland epithelia to complete their life cycle in the mosquito. However, our understanding of the molecular mechanism and the mosquito genes that participate in this process is still very limited. Methodology/Principal Findings We identified an Anopheles gambiae epithelial serine protease (AgESP) that is constitutively expressed in the submicrovillar region of mosquito midgut epithelial cells and in the basal side of the sali...

  4. Living at the edge: biogeographic patterns of habitat segregation conform to speciation by niche expansion in Anopheles gambiae

    OpenAIRE

    Costantini Carlo; Ayala Diego; Guelbeogo Wamdaogo M; Pombi Marco; Some Corentin Y; Bassole Imael HN; Ose Kenji; Fotsing Jean-Marie; Sagnon N'Falé; Fontenille Didier; Besansky Nora J; Simard Frédéric

    2009-01-01

    Abstract Background Ongoing lineage splitting within the African malaria mosquito Anopheles gambiae is compatible with ecological speciation, the evolution of reproductive isolation by divergent natural selection acting on two populations exploiting alternative resources. Divergence between two molecular forms (M and S) identified by fixed differences in rDNA, and characterized by marked, although incomplete, reproductive isolation is occurring in West and Central Africa. To elucidate the rol...

  5. Islands and stepping-stones: comparative population structure of Anopheles gambiae sensu stricto and Anopheles arabiensis in Tanzania and implications for the spread of insecticide resistance.

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    Deodatus Maliti

    Full Text Available Population genetic structures of the two major malaria vectors Anopheles gambiae s.s. and An. arabiensis, differ markedly across Sub-Saharan Africa, which could reflect differences in historical demographies or in contemporary gene flow. Elucidation of the degree and cause of population structure is important for predicting the spread of genetic traits such as insecticide resistance genes or artificially engineered genes. Here the population genetics of An. gambiae s.s. and An. arabiensis in the central, eastern and island regions of Tanzania were compared. Microsatellite markers were screened in 33 collections of female An. gambiae s.l., originating from 22 geographical locations, four of which were sampled in two or three years between 2008 and 2010. An. gambiae were sampled from six sites, An. arabiensis from 14 sites, and both species from two sites, with an additional colonised insectary sample of each species. Frequencies of the knock-down resistance (kdr alleles 1014S and 1014F were also determined. An. gambiae exhibited relatively high genetic differentiation (average pairwise FST = 0.131, significant even between nearby samples, but without clear geographical patterning. In contrast, An. arabiensis exhibited limited differentiation (average FST = 0.015, but strong isolation-by-distance (Mantel test r = 0.46, p = 0.0008. Most time-series samples of An. arabiensis were homogeneous, suggesting general temporal stability of the genetic structure. An. gambiae populations from Dar es Salaam and Bagamoyo were found to have high frequencies of kdr 1014S (around 70%, with almost 50% homozygote but was at much lower frequency on Unguja Island, with no. An. gambiae population genetic differentiation was consistent with an island model of genetic structuring with highly restricted gene flow, contrary to An. arabiensis which was consistent with a stepping-stone model of extensive, but geographically-restricted gene flow.

  6. Ecological niche partitioning between Anopheles gambiae molecular forms in Cameroon: the ecological side of speciation

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    Fotsing Jean-Marie

    2009-05-01

    Full Text Available Abstract Background Speciation among members of the Anopheles gambiae complex is thought to be promoted by disruptive selection and ecological divergence acting on sets of adaptation genes protected from recombination by polymorphic paracentric chromosomal inversions. However, shared chromosomal polymorphisms between the M and S molecular forms of An. gambiae and insufficient information about their relationship with ecological divergence challenge this view. We used Geographic Information Systems, Ecological Niche Factor Analysis, and Bayesian multilocus genetic clustering to explore the nature and extent of ecological and chromosomal differentiation of M and S across all the biogeographic domains of Cameroon in Central Africa, in order to understand the role of chromosomal arrangements in ecological specialisation within and among molecular forms. Results Species distribution modelling with presence-only data revealed differences in the ecological niche of both molecular forms and the sibling species, An. arabiensis. The fundamental environmental envelope of the two molecular forms, however, overlapped to a large extent in the rainforest, where they occurred in sympatry. The S form had the greatest niche breadth of all three taxa, whereas An. arabiensis and the M form had the smallest niche overlap. Correspondence analysis of M and S karyotypes confirmed that molecular forms shared similar combinations of chromosomal inversion arrangements in response to the eco-climatic gradient defining the main biogeographic domains occurring across Cameroon. Savanna karyotypes of M and S, however, segregated along the smaller-scale environmental gradient defined by the second ordination axis. Population structure analysis identified three chromosomal clusters, each containing a mixture of M and S specimens. In both M and S, alternative karyotypes were segregating in contrasted environments, in agreement with a strong ecological adaptive value of

  7. Function and composition of male accessory gland secretions in Anopheles gambiae: a comparison with other insect vectors of infectious diseases

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    Baldini, Francesco; Gabrieli, Paolo; Rogers, David W.; Catteruccia, Flaminia

    2012-01-01

    Human malaria, a major public health burden in tropical and subtropical countries, is transmitted exclusively by the bite of a female Anopheles mosquito. Malaria control strategies aimed at inducing sexual sterility in natural vector populations are an attractive alternative to the use of insecticides. However, despite their importance as disease vectors, limited information is available on the molecular mechanisms regulating fertility in Anopheles mosquitoes. In the major malaria vector, An. gambiae, the full complement of sperm and seminal fluid required for a female’s lifelong egg production is obtained from a single mating event. This single mating has important consequences for the physiology and behavior of An. gambiae females: in particular, they become refractory to further insemination, and they start laying eggs. In other insects including Drosophila, similar post-copulatory changes are induced by seminal proteins secreted by the male accessory glands and transferred to the female during mating. In this review, we analyze the current state of knowledge on the function and characterization of male seminal proteins in An. gambiae, and provide a comparative assessment of the role of these male reproductive factors in other mosquito vectors of human disease in which female post-copulatory behavior has been studied. Knowledge of the factors and mechanisms regulating fertility in An. gambiae and other vectors can help the design of novel control strategies to fight the spread of disease. PMID:22943543

  8. Water quality and immatures of the M and S forms of Anopheles gambiae s.s. and An. arabiensis in a Malian village

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    Touré Yeya T

    2006-04-01

    Full Text Available Abstract Introduction The associations between the immatures of Anopheles gambiae s.s. (Diptera: Culicidae, its M and S forms, and Anopheles arabiensis among and within larval breeding habitats in Banambani, Mali were investigated under varying conditions of water quality and rainfall. The intent was to elucidate on niche partitioning of these taxa. Methods Immatures of An. arabiensis, An. gambiae s.s., and its M and S forms were sampled every alternate day for a month in mid-rainy season from three sampling sites in each of the larval breeding habitats (rock pools, swamp, and puddles. Water quality was characterized by alkalinity, conductivity, dissolved oxygen (D.O., nitrate, orthophosphate, pH, temperature, total dissolved solids (TDS, and turbidity. A type 3 analysis of the GENMOD model was used to examine the associations between the proportional frequencies of young (first and second instar larvae and old (third and fourth instar larvae and pupae or total immatures of species or forms among sampling sites within and among larval breeding habitats during a category of rainfall as influenced by water quality. Results Of the 4,174 immatures sampled, 1,300 were molecularly identified to species and forms. Significant association between the proportional frequencies of young larvae of An. arabiensis, An. gambiae s.s., its M and S forms was found among sampling sites within habitats but not among larval breeding habitats. The proportional frequencies of young larvae of M and S forms varied daily perhaps due to recruitment, mortality, and dispersal within habitats. Conductivity and TDS had significant effects when the proportional frequencies of young larvae of M and S forms among sampling sites within habitats were significantly associated. Alkalinity, D.O., orthophosphate, pH, nitrate, temperature and turbidity had no effects on niche partitioning of species and forms among sampling sites within habitats. Rainfall did not affect the frequencies

  9. Genetic dissection of Anopheles gambiae gut epithelial responses to Serratia marcescens.

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    Stavros Stathopoulos

    2014-03-01

    Full Text Available Genetic variation in the mosquito Anopheles gambiae profoundly influences its ability to transmit malaria. Mosquito gut bacteria are shown to influence the outcome of infections with Plasmodium parasites and are also thought to exert a strong drive on genetic variation through natural selection; however, a link between antibacterial effects and genetic variation is yet to emerge. Here, we combined SNP genotyping and expression profiling with phenotypic analyses of candidate genes by RNAi-mediated silencing and 454 pyrosequencing to investigate this intricate biological system. We identified 138 An. gambiae genes to be genetically associated with the outcome of Serratia marcescens infection, including the peptidoglycan recognition receptor PGRPLC that triggers activation of the antibacterial IMD/REL2 pathway and the epidermal growth factor receptor EGFR. Silencing of three genes encoding type III fibronectin domain proteins (FN3Ds increased the Serratia load and altered the gut microbiota composition in favor of Enterobacteriaceae. These data suggest that natural genetic variation in immune-related genes can shape the bacterial population structure of the mosquito gut with high specificity. Importantly, FN3D2 encodes a homolog of the hypervariable pattern recognition receptor Dscam, suggesting that pathogen-specific recognition may involve a broader family of immune factors. Additionally, we showed that silencing the gene encoding the gustatory receptor Gr9 that is also associated with the Serratia infection phenotype drastically increased Serratia levels. The Gr9 antibacterial activity appears to be related to mosquito feeding behavior and to mostly rely on changes of neuropeptide F expression, together suggesting a behavioral immune response following Serratia infection. Our findings reveal that the mosquito response to oral Serratia infection comprises both an epithelial and a behavioral immune component.

  10. Evolutionary dynamics of the Ty3/gypsy LTR retrotransposons in the genome of Anopheles gambiae.

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    Jose Manuel C Tubio

    Full Text Available Ty3/gypsy elements represent one of the most abundant and diverse LTR-retrotransposon (LTRr groups in the Anopheles gambiae genome, but their evolutionary dynamics have not been explored in detail. Here, we conduct an in silico analysis of the distribution and abundance of the full complement of 1045 copies in the updated AgamP3 assembly. Chromosomal distribution of Ty3/gypsy elements is inversely related to arm length, with densities being greatest on the X, and greater on the short versus long arms of both autosomes. Taking into account the different heterochromatic and euchromatic compartments of the genome, our data suggest that the relative abundance of Ty3/gypsy LTRrs along each chromosome arm is determined mainly by the different proportions of heterochromatin, particularly pericentric heterochromatin, relative to total arm length. Additionally, the breakpoint regions of chromosomal inversion 2La appears to be a haven for LTRrs. These elements are underrepresented more than 7-fold in euchromatin, where 33% of the Ty3/gypsy copies are associated with genes. The euchromatin on chromosome 3R shows a faster turnover rate of Ty3/gypsy elements, characterized by a deficit of proviral sequences and the lowest average sequence divergence of any autosomal region analyzed in this study. This probably reflects a principal role of purifying selection against insertion for the preservation of longer conserved syntenyc blocks with adaptive importance located in 3R. Although some Ty3/gypsy LTRrs show evidence of recent activity, an important fraction are inactive remnants of relatively ancient insertions apparently subject to genetic drift. Consistent with these computational predictions, an analysis of the occupancy rate of putatively older insertions in natural populations suggested that the degenerate copies have been fixed across the species range in this mosquito, and also are shared with the sibling species Anopheles arabiensis.

  11. INSECTICIDAL ACTIVITIES OF ESSENTIAL OILS EXTRACTED FROM THREE SPECIES OF POACEAE ON ANOPHELES GAMBIAE SPP, MAJOR VECTOR OF MALARIA

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    Dominique C. K. Sohounhloué

    2010-12-01

    Full Text Available In this paper, the insecticidal activities on Anopheles gambiae spp of the essential oils (EO extracted from the dry leaves of some species collected in Benin were studied. The essential oil yields are 2.8, 1.7 and 1.4�0respectively for Cymbopogon schoanenthus (L. Spreng (CS, Cymbopogon citratus Stapf. (CC and Cymbopogon giganteus (Hochst. Chiov (CG. The GC/MS analysis showed that the EO of CS had a larger proportion in oxygenated monoterpenes (86.3�20whereas those of the sheets of CC and CG are relatively close proportions (85.5�0and 82.7�0respectively with. The piperitone (68.5�  2-carene (11.5� and -eudesmol (4.6�20are the major components of the EO of CS while trans para-mentha-1(7,8-dien-2-ol (31.9� trans para-mentha-2,8-dien-1-ol (19.6� cis para-mentha-2,8-dien-1-ol (7.2� trans piperitol (6.3�20and limonene (6.3�20prevailed in the EO of CG. The EO of CC revealed a rich composition in geranial (41.3� neral (33� myrcene (10.4� and geraniol (6.6� The biological tests have shown that these three EO induced 100�0mortality of Anopheles gambiae to 1.1, 586.58 and 1549 µg•cm-2 respectively for CC, CS and CG. These effects are also illustrated by weak lethal concentration for 50�0anopheles population (CC: 0.306; CS: 152.453 and CG: 568.327 µg•cm-2 in the same order of reactivity. The EO of CC appeared most active on two stocks (sensitive and resistant of Anopheles gambiae.

  12. Mating activates the heme peroxidase HPX15 in the sperm storage organ to ensure fertility in Anopheles gambiae

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    Shaw, W. Robert; Teodori, Eleonora; Mitchell, Sara N.; Baldini, Francesco; Gabrieli, Paolo; Rogers, David W.; Catteruccia, Flaminia

    2014-01-01

    Anopheles gambiae mosquitoes are major African vectors of malaria, a disease that kills more than 600,000 people every year. Given the spread of insecticide resistance in natural mosquito populations, alternative vector control strategies aimed at reducing the reproductive success of mosquitoes are being promoted. Unlike many other insects, An. gambiae females mate a single time in their lives and must use sperm stored in the sperm storage organ, the spermatheca, to fertilize a lifetime's supply of eggs. Maintenance of sperm viability during storage is therefore crucial to the reproductive capacity of these mosquitoes. However, to date, no information is available on the factors and mechanisms ensuring sperm functionality in the spermatheca. Here we identify cellular components and molecular mechanisms used by An. gambiae females to maximize their fertility. Pathways of energy metabolism, cellular transport, and oxidative stress are strongly regulated by mating in the spermatheca. We identify the mating-induced heme peroxidase (HPX) 15 as an important factor in long-term fertility, and demonstrate that its function is required during multiple gonotrophic cycles. We find that HPX15 induction is regulated by sexually transferred 20-hydroxy-ecdysone (20E), a steroid hormone that is produced by the male accessory glands and transferred during copulation, and that expression of this peroxidase is mediated via the 20E nuclear receptor. To our knowledge, our findings provide the first evidence of the mechanisms regulating fertility in Anopheles, and identify HPX15 as a target for vector control. PMID:24711401

  13. Larvicidal activity of metabolites from the endophytic Podospora sp. against the malaria vector Anopheles gambiae.

    Science.gov (United States)

    Matasyoh, Josphat C; Dittrich, Birger; Schueffler, Anja; Laatsch, Hartmut

    2011-03-01

    In a screening for natural products with mosquito larvicidal activities, the endophytic fungus Podospora sp. isolated from the plant Laggera alata (Asteraceae) was conspicuous. Two xanthones, sterigmatocystin (1) and secosterigmatocystin (2), and an anthraquinone derivative (3) 13-hydroxyversicolorin B were isolated after fermentation on M(2) medium. These compounds were characterised using spectroscopic and X-ray analysis and examined against third instar larvae of Anopheles gambiae. The results demonstrated that compound 1 was the most potent one with LC(50) and LC(90) values of 13.3 and 73.5 ppm, respectively. Over 95% mortality was observed at a concentration 100 ppm after 24 h. These results compared farvorably with the commercial larvicide pylarvex® that showed 100% mortality at the same concentration. Compound 3 was less potent and had an LC(50) of 294.5 ppm and over 95% mortality was achieved at a concentration of 1,000 ppm. Secosterigmatocystin (2) revealed relatively weak activity and therefore LC values were not determined.

  14. Authentication scheme for routine verification of genetically similar laboratory colonies: a trial with Anopheles gambiae

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    Sutcliffe Alice C

    2009-10-01

    Full Text Available Abstract Background When rearing morphologically indistinguishable laboratory strains concurrently, the threat of unintentional genetic contamination is constant. Avoidance of accidental mixing of strains is difficult due to the use of common equipment, technician error, or the possibility of self relocation by adult mosquitoes ("free fliers". In many cases, laboratory strains are difficult to distinguish because of morphological and genetic similarity, especially when laboratory colonies are isolates of certain traits from the same parental strain, such as eye color mutants, individuals with certain chromosomal arrangements or high levels of insecticide resistance. Thus, proving genetic integrity could seem incredibly time-consuming or impossible. On the other hand, lacking proof of genetically isolated laboratory strains could question the validity of research results. Results We present a method for establishing authentication matrices to routinely distinguish and confirm that laboratory strains have not become physically or genetically mixed through contamination events in the laboratory. We show a specific example with application to Anopheles gambiae sensu stricto strains at the Malaria Research and Reference Reagent Resource Center. This authentication matrix is essentially a series of tests yielding a strain-specific combination of results. Conclusion These matrix-based methodologies are useful for several mosquito and insect populations but must be specifically tailored and altered for each laboratory based on the potential contaminants available at any given time. The desired resulting authentication plan would utilize the least amount of routine effort possible while ensuring the integrity of the strains.

  15. Characterization of Anopheles gambiae Transglutaminase 3 (AgTG3) and Its Native Substrate Plugin*

    Science.gov (United States)

    Le, Binh V.; Nguyen, Jennifer B.; Logarajah, Shankar; Wang, Bo; Marcus, Jacob; Williams, Hazel P.; Catteruccia, Flaminia; Baxter, Richard H. G.

    2013-01-01

    Male Anopheles mosquitoes coagulate their seminal fluids via cross-linking of a substrate, called Plugin, by the seminal transglutaminase AgTG3. Formation of the “mating plug” by cross-linking Plugin is necessary for efficient sperm storage by females. AgTG3 has a similar degree of sequence identity (∼30%) to both human Factor XIII (FXIII) and tissue transglutaminase 2 (hTG2). Here we report the solution structure and in vitro activity for the cross-linking reaction of AgTG3 and Plugin. AgTG3 is a dimer in solution and exhibits Ca2+-dependent nonproteolytic activation analogous to cytoplasmic FXIII. The C-terminal domain of Plugin is predominantly α-helical with extended tertiary structure and oligomerizes in solution. The specific activity of AgTG3 was measured as 4.25 × 10−2 units mg−1. AgTG3 is less active than hTG2 assayed using the general substrate TVQQEL but has 8–10× higher relative activity when Plugin is the substrate. Mass spectrometric analysis of cross-linked Plugin detects specific peptides including a predicted consensus motif for cross-linking by AgTG3. These results support the development of AgTG3 inhibitors as specific and effective chemosterilants for A. gambiae. PMID:23288850

  16. Carbamate and Pyrethroid Resistance in the Akron Strain of Anopheles gambiae

    Science.gov (United States)

    Mutunga, James M.; Anderson, Troy D.; Craft, Derek T.; Gross, Aaron D.; Swale, Daniel R.; Tong, Fan; Wong, Dawn M.; Carlier, Paul R.; Bloomquist, Jeffrey R.

    2015-01-01

    Insecticide resistance in the malaria vector, Anopheles gambiae is a serious problem, epitomized by the multi-resistant Akron strain, originally isolated in the country of Benin. Here we report resistance in this strain to pyrethroids and DDT (13-fold to 35-fold compared to the susceptible G3 strain), but surprisingly little resistance to etofenprox, a compound sometimes described as a “pseudo-pyrethroid.” There was also strong resistance to topically-applied commercial carbamates (45-fold to 81-fold), except for the oximes aldicarb and methomyl. Biochemical assays showed enhanced cytochrome P450 monooxygenase and carboxylesterase activity, but not that of glutathione-S-transferase. A series of substituted α,α,α,-trifluoroacetophenone oxime methylcarbamates were evaluated for enzyme inhibition potency and toxicity against G3 and Akron mosquitoes. The compound bearing an unsubstituted phenyl ring showed the greatest toxicity to mosquitoes of both strains. Low cross resistance in Akron was retained by all analogs in the series. Kinetic analysis of acetylcholinesterase activity and its inhibition by insecticides in the G3 strain showed inactivation rate constants greater than that of propoxur, and against Akron enzyme inactivation rate constants similar to that of aldicarb. However, inactivation rate constants against recombinant human AChE were essentially identical to that of the G3 strain. Thus, the acetophenone oxime carbamates described here, though potent insecticides that control resistant Akron mosquitoes, require further structural modification to attain acceptable selectivity and human safety. PMID:26047119

  17. Gene expression patterns associated with blood-feeding in the malaria mosquito Anopheles gambiae

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    Hogan James R

    2005-01-01

    Full Text Available Abstract Background Blood feeding, or hematophagy, is a behavior exhibited by female mosquitoes required both for reproduction and for transmission of pathogens. We determined the expression patterns of 3,068 ESTs, representing ~2,000 unique gene transcripts using cDNA microarrays in adult female Anopheles gambiae at selected times during the first two days following blood ingestion, at 5 and 30 min during a 40 minute blood meal and at 0, 1, 3, 5, 12, 16, 24 and 48 hours after completion of the blood meal and compared their expression to transcript levels in mosquitoes with access only to a sugar solution. Results In blood-fed mosquitoes, 413 unique transcripts, approximately 25% of the total, were expressed at least two-fold above or below their levels in the sugar-fed mosquitoes, at one or more time points. These differentially expressed gene products were clustered using k-means clustering into Early Genes, Middle Genes, and Late Genes, containing 144, 130, and 139 unique transcripts, respectively. Several genes from each group were analyzed by quantitative real-time PCR in order to validate the microarray results. Conclusion The expression patterns and annotation of the genes in these three groups (Early, Middle, and Late genes are discussed in the context of female mosquitoes' physiological responses to blood feeding, including blood digestion, peritrophic matrix formation, egg development, and immunity.

  18. Detection of the East and West African kdr mutation in Anopheles gambiae and Anopheles arabiensis from Uganda using a new assay based on FRET/Melt Curve analysis

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    Backeljau Thierry

    2006-02-01

    Full Text Available Abstract Background Appropriate monitoring of vector resistance to insecticides is an integral component of planning and evaluation of insecticide use in malaria control programmes. The malaria vectors Anopheles gambiae s.s. and Anopheles arabiensis have developed resistance to pyrethroid insecticides as a result of a mechanism conferring reduced nervous system sensitivity, better known as knockdown resistance (kdr. In An. gambiae s.s. and An. arabiensis, two different substitutions in the para-type sodium channel, a L1014F substitution common in West Africa and a L1014S replacement found in Kenya, are linked with kdr. Two different allele-specific polymerase chain reactions (AS-PCR are needed to detect these known kdr mutations. However, these AS-PCR assays rely on a single nucleotide polymorphism mismatch, which can result in unreliable results. Methods Here, a new assay for the detection of knockdown resistance in An. gambiae s.s. and An. arabiensis based on Fluorescence Resonance Energy Transfer/Melt Curve analysis (FRET/MCA is presented and compared with the existing assays. Results The new FRET/MCA method has the important advantage of detecting both kdr alleles in one assay. Moreover, results show that the FRET/MCA is more reliable and more sensitive than the existing AS-PCR assays and is able to detect new genotypes. By using this technique, the presence of the East African kdr mutation (L1014S is shown for the first time in An. arabiensis specimens from Uganda. In addition, a new kdr genotype is reported in An. gambiae s.s. from Uganda, where four An. gambiae s.s. mosquitoes possess both, the West (L1014F and East (L1014S African kdr allele, simultaneously. Conclusion The presence of both kdr mutations in the same geographical region shows the necessity of a reliable assay that enables to detect both mutations in one single assay. Hence, this new assay based on FRET/MCA will improve the screening of the kdr frequencies in An. gambiae s

  19. Toxicity of essential oil from Indian borage on the larvae of the African malaria vector mosquito, Anopheles gambiae

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    Kweka Eliningaya J

    2012-12-01

    Full Text Available Abstract Background Essential oils are currently studied for the control of different disease vectors, because of their efficacy on targeted organisms. In the present investigation, the larvicidal potential of essential oil extracted from Indian borage (Plectranthus amboinicus was studied against the African anthropophagic malaria vector mosquito, Anopheles gambiae. The larvae of An. gambiae s.s laboratory colony and An. gambiae s.l of wild populations were assayed and the larval mortality was observed at 12, 24 and 48 h after exposure period with the concentrations of 3.125, 6.25, 12.5, 25, 50 and 100 ppm. Findings Larval mortality rates of the essential oil was entirely time and dose dependent. The LC50 values of the laboratory colony were 98.56 (after 12h 55.20 (after 24 h and 32.41 ppm (after 48 h and the LC90 values were 147.40 (after 12h, 99.09 (after 24 h and 98.84 ppm (after 48 h. The LC50 and LC90 values of the wild population were 119.52, 179.85 (after 12h 67.53, 107.60 (after 24 h and 25.51, 111.17 ppm (after 48 h respectively. The oil showed good larvicidal potential after 48 h of exposure period against An. gambiae. The essential oil of Indian borage is a renowned natural source of larvicides for the control of the African malaria vector mosquito, An. gambiae. Conclusion The larvicidal efficacy shown by plant extracts against An. gambiae should be tested in semi field and small scale trials for effective compounds to supplement the existing larval control tools.

  20. Select small core structure carbamates exhibit high contact toxicity to "carbamate-resistant" strain malaria mosquitoes, Anopheles gambiae (Akron.

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    Dawn M Wong

    Full Text Available Acetylcholinesterase (AChE is a proven target for control of the malaria mosquito (Anopheles gambiae. Unfortunately, a single amino acid mutation (G119S in An. gambiae AChE-1 (AgAChE confers resistance to the AChE inhibitors currently approved by the World Health Organization for indoor residual spraying. In this report, we describe several carbamate inhibitors that potently inhibit G119S AgAChE and that are contact-toxic to carbamate-resistant An. gambiae. PCR-RFLP analysis was used to confirm that carbamate-susceptible G3 and carbamate-resistant Akron strains of An. gambiae carry wild-type (WT and G119S AChE, respectively. G119S AgAChE was expressed and purified for the first time, and was shown to have only 3% of the turnover number (k(cat of the WT enzyme. Twelve carbamates were then assayed for inhibition of these enzymes. High resistance ratios (>2,500-fold were observed for carbamates bearing a benzene ring core, consistent with the carbamate-resistant phenotype of the G119S enzyme. Interestingly, resistance ratios for two oxime methylcarbamates, and for five pyrazol-4-yl methylcarbamates were found to be much lower (4- to 65-fold. The toxicities of these carbamates to live G3 and Akron strain An. gambiae were determined. As expected from the enzyme resistance ratios, carbamates bearing a benzene ring core showed low toxicity to Akron strain An. gambiae (LC(50>5,000 μg/mL. However, one oxime methylcarbamate (aldicarb and five pyrazol-4-yl methylcarbamates (4a-e showed good to excellent toxicity to the Akron strain (LC(50 = 32-650 μg/mL. These results suggest that appropriately functionalized "small-core" carbamates could function as a resistance-breaking anticholinesterase insecticides against the malaria mosquito.

  1. Identification and expression profiling of putative odorant-binding proteins in the malaria mosquitoes, Anopheles gambiae and A.arabiensis

    Institute of Scientific and Technical Information of China (English)

    LI Zhengxi; Jing-Jiang ZHOU; SHEN Zuorui; Lin FIELD

    2004-01-01

    Olfaction plays a major role in host-seeking behaviour of mosquitoes. An informatics-based genome-wide analysis of odorant-binding protein (OBP) homologues is undertaken,and 32 putative OBP genes in total in the whole genome sequences of Anopheles gambiae are identified. Tissue-specific expression patterns of all A. gambiae OBP candidates are determined by semi-quantitative Reverse Transcription (RT)-PCR using mosquito actin gene as internal expression control standard. The results showed that 20 OBP candidates had strong expression in mosquito olfactory tissues (female antennae), which indicate that OBPs may play an important role in regulating mosquito olfactory behaviours. Species-specific expression patterns of all putative anopheline OBPs are also studied in two of the most important malaria vectors in A. gambiae complex, i.e.A. gambiae and A. arabiensis, which found 12 of the putative OBP genes examined displayed species-differential expression patterns. The cumulative relative expression intensity of the OBPs in A. arabiensis antennae was higher than that in A. gambiae (the ratio is 1441.45:1314.12), which might be due to their different host preference behaviour. While A.gambiae is a highly anthropophilic mosquito, A. arabiensis is more opportunistic (varying from anthropophilic to zoophilic). So the latter should need more OBPs to support its host selection preference. Identification of mosquito OBPs and verification of their tissue- and species-specific expression patterns represent the first step towards further molecular analysis of mosquito olfactory mechanism, such as recombinant expression and ligand identification.

  2. Effects of bed net use, female size, and plant abundance on the first meal choice (blood vs sugar) of the malaria mosquito Anopheles gambiae

    OpenAIRE

    Stone Chris M; Jackson Bryan T; Foster Woodbridge A

    2012-01-01

    Abstract Background The purpose of this study was to determine whether the sugar-or-blood meal choice of Anopheles gambiae females one day after emergence is influenced by blood-host presence and accessibility, nectariferous plant abundance, and female size. This tested the hypothesis that the initial meal of female An. gambiae is sugar, even when a blood host is available throughout the night, and, if not, whether the use of a bed net diverts mosquitoes to sugar sources. Methods Females and ...

  3. A CRISPR-Cas9 Gene Drive System Targeting Female Reproduction in the Malaria Mosquito vector Anopheles gambiae

    OpenAIRE

    Hammond, Andrew; Galizi, Roberto; Kyrou, Kyros; Simoni, Alekos; Siniscalchi, Carla; Katsanos, Dimitris; Gribble, Matthew; Baker, Dean; Marois, Eric; Russell, Steven; Burt, Austin; Windbichler, Nikolai; Crisanti, Andrea; Nolan, Tony

    2015-01-01

    Gene-drive systems that enable super-Mendelian inheritance of a transgene have the potential to modify insect populations over a timeframe of a few years [AU please provide a real estimate, this seems vague]. We describe CRISPR-Cas9 endonuclease constructs that function as gene-drive systems in Anopheles gambiae, the main vector for malaria [AU:OK?]. We identified three genes (AGAP005958, AGAP011377 and AGAP007280) that confer a recessive female sterility phenotype upon disruption, and insert...

  4. Some strains of Plasmodium falciparum, a human malaria parasite, evade the complement-like system of Anopheles gambiae mosquitoes

    OpenAIRE

    Molina-Cruz, Alvaro; DeJong, Randall J.; Ortega, Corrie; Haile, Ashley; Abban, Ekua; Rodrigues, Janneth; Jaramillo-Gutierrez, Giovanna; Barillas-Mury, Carolina

    2012-01-01

    Plasmodium falciparum lines differ in their ability to infect mosquitoes. The Anopheles gambiae L3-5 refractory (R) line melanizes most Plasmodium species, including the Brazilian P. falciparum 7G8 line, but it is highly susceptible to some African P. falciparum strains such as 3D7, NF54, and GB4. We investigated whether these lines differ in their ability to evade the mosquito immune system. Silencing key components of the mosquito complement-like system [thioester-containing protein 1 (TEP1...

  5. Anopheles gambiae Ag-STAT, a new insect member of the STAT family, is activated in response to bacterial infection.

    OpenAIRE

    Barillas-Mury, C; Han, Y S; Seeley, D; Kafatos, F C

    1999-01-01

    A new insect member of the STAT family of transcription factors (Ag-STAT) has been cloned from the human malaria vector Anopheles gambiae. The domain involved in DNA interaction and the SH2 domain are well conserved. Ag-STAT is most similar to Drosophila D-STAT and to vertebrate STATs 5 and 6, constituting a proposed ancient class A of the STAT family. The mRNA is expressed at all developmental stages, and the protein is present in hemocytes, pericardial cells, midgut, skeletal muscle and fat...

  6. Distribution of knock-down resistance mutations in Anopheles gambiae molecular forms in west and west-central Africa

    Directory of Open Access Journals (Sweden)

    Caccone Adalgisa

    2008-04-01

    Full Text Available Abstract Background Knock-down resistance (kdr to DDT and pyrethroids in the major Afrotropical vector species, Anopheles gambiae sensu stricto, is associated with two alternative point mutations at amino acid position 1014 of the voltage-gated sodium channel gene, resulting in either a leucine-phenylalanine (L1014F, or a leucine-serine (L1014S substitution. In An. gambiae S-form populations, the former mutation appears to be widespread in west Africa and has been recently reported from Uganda, while the latter, originally recorded in Kenya, has been recently found in Gabon, Cameroon and Equatorial Guinea. In M-form populations surveyed to date, only the L1014F mutation has been found, although less widespread and at lower frequencies than in sympatric S-form populations. Methods Anopheles gambiae M- and S-form specimens from 19 sites from 11 west and west-central African countries were identified to molecular form and genotyped at the kdr locus either by Hot Oligonucleotide Ligation Assay (HOLA or allele-specific PCR (AS-PCR. Results The kdr genotype was determined for about 1,000 An. gambiae specimens. The L1014F allele was found at frequencies ranging from 6% to 100% in all S-form samples (N = 628, with the exception of two samples from Angola, where it was absent, and coexisted with the L1014S allele in samples from Cameroon, Gabon and north-western Angola. The L1014F allele was present in M-form samples (N = 354 from Benin, Nigeria, and Cameroon, where both M- and S-forms were sympatric. Conclusion The results represent the most comprehensive effort to analyse the overall distribution of the L1014F and L1014S mutations in An. gambiae molecular forms, and will serve as baseline data for resistance monitoring. The overall picture shows that the emergence and spread of kdr alleles in An. gambiae is a dynamic process and that there is marked intra- and inter-form heterogeneity in resistance allele frequencies. Further studies are needed to

  7. Biochemical characterization of Anopheles gambiae SRPN6, a malaria parasite invasion marker in mosquitoes.

    Directory of Open Access Journals (Sweden)

    Chunju An

    Full Text Available Serine proteinase inhibitors of the serpin family are well known as negative regulators of hemostasis, thrombolysis and innate immune responses. Additionally, non-inhibitory serpins serve functions as chaperones, hormone transporters, or anti-angiogenic factors. In the African malaria mosquito, Anopheles gambiae s.s., at least three serpins (SRPNs are implicated in the innate immune response against malaria parasites. Based on reverse genetic and cell biological analyses, AgSRPN6 limits parasite numbers and transmission and has been postulated to control melanization and complement function in mosquitoes. This study aimed to characterize AgSRPN6 biophysically and determine its biochemical mode of action. The structure model of AgSRPN6, as predicted by I-Tasser showed the protein in the native serpin fold, with three central β-sheets, nine surrounding α-helices, and a protruding reactive center loop. This structure is in agreement with biophysical and functional data obtained from recombinant (r AgSRPN6, produced in Escherichia coli. The physical properties of purified rAgSRPN6 were investigated by means of analytical ultracentrifugation, circular dichroism, and differential scanning calorimetry tools. The recombinant protein exists predominantly as a monomer in solution, is composed of a mixture of α-helices and β-sheets, and has a mid-point unfolding temperature of 56°C. Recombinant AgSRPN6 strongly inhibited porcine pancreatic kallikrein and to a lesser extent bovine pancreatic trypsin in vitro. Furthermore, rAgSRPN6 formed inhibitory, SDS-stable, higher molecular weight complexes with prophenoloxidase-activating proteinase (PAP1, PAP3, and Hemolymph protein (HP6, which are required for melanization in the lepidopteran model organism, Manduca sexta. Taken together, our results strongly suggest that AgSRPN6 takes on a native serpin fold and is an inhibitor of trypsin-like serine proteinases.

  8. Molecular evolution of immune genes in the malaria mosquito Anopheles gambiae.

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    Tovi Lehmann

    Full Text Available BACKGROUND: As pathogens that circumvent the host immune response are favoured by selection, so are host alleles that reduce parasite load. Such evolutionary processes leave their signature on the genes involved. Deciphering modes of selection operating on immune genes might reveal the nature of host-pathogen interactions and factors that govern susceptibility in host populations. Such understanding would have important public health implications. METHODOLOGY/FINDINGS: We analyzed polymorphisms in four mosquito immune genes (SP14D1, GNBP, defensin, and gambicin to decipher selection effects, presumably mediated by pathogens. Using samples of Anopheles arabiensis, An. quadriannulatus and four An. gambiae populations, as well as published sequences from other Culicidae, we contrasted patterns of polymorphisms between different functional units of the same gene within and between populations. Our results revealed selection signatures operating on different time scales. At the most recent time scale, within-population diversity revealed purifying selection. Between populations and between species variation revealed reduced differentiation (GNBP and gambicin at coding vs. noncoding- regions, consistent with balancing selection. McDonald-Kreitman tests between An. quadriannulatus and both sibling species revealed higher fixation rate of synonymous than nonsynonymous substitutions (GNBP in accordance with frequency dependent balancing selection. At the longest time scale (>100 my, PAML analysis using distant Culicid taxa revealed positive selection at one codon in gambicin. Patterns of genetic variation were independent of exposure to human pathogens. SIGNIFICANCE AND CONCLUSIONS: Purifying selection is the most common form of selection operating on immune genes as it was detected on a contemporary time scale on all genes. Selection for "hypervariability" was not detected, but negative balancing selection, detected at a recent evolutionary time scale

  9. Biochemical characterization of Anopheles gambiae SRPN6, a malaria parasite invasion marker in mosquitoes.

    Science.gov (United States)

    An, Chunju; Hiromasa, Yasuaki; Zhang, Xin; Lovell, Scott; Zolkiewski, Michal; Tomich, John M; Michel, Kristin

    2012-01-01

    Serine proteinase inhibitors of the serpin family are well known as negative regulators of hemostasis, thrombolysis and innate immune responses. Additionally, non-inhibitory serpins serve functions as chaperones, hormone transporters, or anti-angiogenic factors. In the African malaria mosquito, Anopheles gambiae s.s., at least three serpins (SRPNs) are implicated in the innate immune response against malaria parasites. Based on reverse genetic and cell biological analyses, AgSRPN6 limits parasite numbers and transmission and has been postulated to control melanization and complement function in mosquitoes. This study aimed to characterize AgSRPN6 biophysically and determine its biochemical mode of action. The structure model of AgSRPN6, as predicted by I-Tasser showed the protein in the native serpin fold, with three central β-sheets, nine surrounding α-helices, and a protruding reactive center loop. This structure is in agreement with biophysical and functional data obtained from recombinant (r) AgSRPN6, produced in Escherichia coli. The physical properties of purified rAgSRPN6 were investigated by means of analytical ultracentrifugation, circular dichroism, and differential scanning calorimetry tools. The recombinant protein exists predominantly as a monomer in solution, is composed of a mixture of α-helices and β-sheets, and has a mid-point unfolding temperature of 56°C. Recombinant AgSRPN6 strongly inhibited porcine pancreatic kallikrein and to a lesser extent bovine pancreatic trypsin in vitro. Furthermore, rAgSRPN6 formed inhibitory, SDS-stable, higher molecular weight complexes with prophenoloxidase-activating proteinase (PAP)1, PAP3, and Hemolymph protein (HP)6, which are required for melanization in the lepidopteran model organism, Manduca sexta. Taken together, our results strongly suggest that AgSRPN6 takes on a native serpin fold and is an inhibitor of trypsin-like serine proteinases. PMID:23152794

  10. Multicopper oxidase-3 is a laccase associated with the peritrophic matrix of Anopheles gambiae.

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    Minglin Lang

    Full Text Available The multicopper oxidase (MCO family of enzymes includes laccases, which oxidize a broad range of substrates including polyphenols and phenylendiamines; ferroxidases, which oxidize ferrous iron; and several other oxidases with specific substrates such as ascorbate, bilirubin or copper. The genome of Anopheles gambiae, a species of mosquito, encodes five putative multicopper oxidases. Of these five, only AgMCO2 has known enzymatic and physiological functions: it is a highly conserved laccase that functions in cuticle pigmentation and tanning by oxidizing dopamine and dopamine derivatives. AgMCO3 is a mosquito-specific gene that is expressed predominantly in adult midguts and Malpighian tubules. To determine its enzymatic function, we purified recombinant AgMCO3 and analyzed its activity. AgMCO3 oxidized hydroquinone (a p-diphenol, the five o-diphenols tested, 2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid (ABTS, and p-phenylenediamine, but not ferrous iron. The catalytic efficiencies of AgMCO3 were similar to those of cuticular laccases (MCO2 orthologs, except that AgMCO3 oxidized all of the phenolic substrates with similar efficiencies whereas the MCO2 isoforms were less efficient at oxidizing catechol or dopa. These results demonstrate that AgMCO3 can be classified as a laccase and suggest that AgMCO3 has a somewhat broader substrate specificity than MCO2 orthologs. In addition, we observed AgMCO3 immunoreactivity in the peritrophic matrix, which functions as a selective barrier between the blood meal and midgut epithelial cells, protecting the midgut from mechanical damage, pathogens, and toxic molecules. We propose that AgMCO3 may oxidize toxic molecules in the blood meal leading to detoxification or to cross-linking of the molecules to the peritrophic matrix, thus targeting them for excretion.

  11. Bacteria- and IMD pathway-independent immune defenses against Plasmodium falciparum in Anopheles gambiae.

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    Benjamin J Blumberg

    Full Text Available The mosquito Anopheles gambiae uses its innate immune system to control bacterial and Plasmodium infection of its midgut tissue. The activation of potent IMD pathway-mediated anti-Plasmodium falciparum defenses is dependent on the presence of the midgut microbiota, which activate this defense system upon parasite infection through a peptidoglycan recognition protein, PGRPLC. We employed transcriptomic and reverse genetic analyses to compare the P. falciparum infection-responsive transcriptomes of septic and aseptic mosquitoes and to determine whether bacteria-independent anti-Plasmodium defenses exist. Antibiotic treated aseptic mosquitoes mounted molecular immune responses representing a variety of immune functions upon P. falciparum infection. Among other immune factors, our analysis uncovered a serine protease inhibitor (SRPN7 and Clip-domain serine protease (CLIPC2 that were transcriptionally induced in the midgut upon P. falciparum infection, independent of bacteria. We also showed that SRPN7 negatively and CLIPC2 positively regulate the anti-Plasmodium defense, independently of the midgut-associated bacteria. Co-silencing assays suggested that these two genes may function together in a signaling cascade. Neither gene was regulated, nor modulated, by infection with the rodent malaria parasite Plasmodium berghei, suggesting that SRPN7 and CLIPC2 are components of a defense system with preferential activity towards P. falciparum. Further analysis using RNA interference determined that these genes do not regulate the anti-Plasmodium defense mediated by the IMD pathway, and both factors act as agonists of the endogenous midgut microbiota, further demonstrating the lack of functional relatedness between these genes and the bacteria-dependent activation of the IMD pathway. This is the first study confirming the existence of a bacteria-independent, anti-P. falciparum defense. Further exploration of this anti-Plasmodium defense will help clarify

  12. Cladistic analysis of the subgenus Anopheles (Anopheles) Meigen (Diptera: Culicidae) based on morphological characters.

    Science.gov (United States)

    Collucci, Eliana; Sallum, Maria Anice Mureb

    2007-06-01

    In the present study, we used morphological characters to estimate phylogenetic relationships among members of the subgenus Anopheles Meigen. Phylogenetic analyses were carried out for 36 species of Anopheles (Anopheles). An. (Stethomyia) kompi Edwards, An. (Lophopodomyia) gilesi (Peryassú), Bironella hollandi Taylor, An. (Nyssorhynchus) oswaldoi (Peryassú) and An. (Cellia) maculatus Theobald were employed as outgroups. One hundred one characters of the external morphology of the adult male, adult female, fourth-instar larva, and pupa were scored and analyzed under the parsimony criterion in PAUP. Phylogenetic relationships among the series and several species informal groups of Anopheles (Anopheles) were hypothesized. The results suggest that Anopheles (Anopheles) is monophyletic. Additionally, most species groups included in the analysis were demonstrated to be monophyletic.

  13. Cladistic analysis of the subgenus Anopheles (Anopheles Meigen (Diptera: Culicidae based on morphological characters

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    Eliana Collucci

    2007-06-01

    Full Text Available In the present study, we used morphological characters to estimate phylogenetic relationships among members of the subgenus Anopheles Meigen. Phylogenetic analyses were carried out for 36 species of Anopheles (Anopheles. An. (Stethomyia kompi Edwards, An. (Lophopodomyia gilesi (Peryassú, Bironella hollandi Taylor, An. (Nyssorhynchus oswaldoi (Peryassú and An. (Cellia maculatus Theobald were employed as outgroups. One hundred one characters of the external morphology of the adult male, adult female, fourth-instar larva, and pupa were scored and analyzed under the parsimony criterion in PAUP. Phylogenetic relationships among the series and several species informal groups of Anopheles (Anopheles were hypothesized. The results suggest that Anopheles (Anopheles is monophyletic. Additionally, most species groups included in the analysis were demonstrated to be monophyletic.

  14. The effect of aliphatic carboxylic acids on olfaction-based host-seeking of the malaria mosquito Anopheles gambiae sensu stricto

    NARCIS (Netherlands)

    Smallegange, R.C.; Qiu, Y.T.; Bukovinszkine-Kiss, G.; Loon, van J.J.A.; Takken, W.

    2009-01-01

    The role of aliphatic carboxylic acids in host-seeking response of the malaria mosquito Anopheles gambiae sensu stricto was examined both in a dual-choice olfactometer and with indoor traps. A basic attractive blend of ammonia + lactic acid served as internal standard odor. Single carboxylic acids w

  15. Effects of blood-feeding on olfactory sensitivity of the malaria mosquito Anopheles gambiae: application of mixed linear models to account for repeated measurements

    NARCIS (Netherlands)

    Qiu, Y.T.; Gort, G.; Torricelli, A.; Takken, W.; Loon, van J.J.A.

    2013-01-01

    Olfaction plays an important role in the host-seeking behavior of the malaria mosquito Anopheles gambiae. After a complete blood meal, female mosquitoes will not engage in host-seeking behavior until oviposition has occurred. We investigated if peripheral olfactory sensitivity changed after a blood

  16. Evaluation of two counterflow traps for testing behaviour-mediating compounds for the malaria vector Anopheles gambiae s.s. under semi-field conditions in Tanzania

    NARCIS (Netherlands)

    Schmied, W.H.; Takken, W.; Killeen, G.F.; Knols, B.G.J.; Smallegange, R.C.

    2008-01-01

    Background Evaluation of mosquito responses towards different trap-bait combinations in field trials is a time-consuming process that can be shortened by experiments in contained semi-field systems. Possible use of the BG Sentinel (BGS) trap to sample Anopheles gambiae s.s. was evaluated. The effici

  17. Sugar-fermenting yeast as an organic source of carbon dioxide to attract the malaria mosquito Anopheles gambiae

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    Spitzen Jeroen

    2010-10-01

    Full Text Available Abstract Background Carbon dioxide (CO2 plays an important role in the host-seeking process of opportunistic, zoophilic and anthropophilic mosquito species and is, therefore, commonly added to mosquito sampling tools. The African malaria vector Anopheles gambiae sensu stricto is attracted to human volatiles augmented by CO2. This study investigated whether CO2, usually supplied from gas cylinders acquired from commercial industry, could be replaced by CO2 derived from fermenting yeast (yeast-produced CO2. Methods Trapping experiments were conducted in the laboratory, semi-field and field, with An. gambiae s.s. as the target species. MM-X traps were baited with volatiles produced by mixtures of yeast, sugar and water, prepared in 1.5, 5 or 25 L bottles. Catches were compared with traps baited with industrial CO2. The additional effect of human odours was also examined. In the laboratory and semi-field facility dual-choice experiments were conducted. The effect of traps baited with yeast-produced CO2 on the number of mosquitoes entering an African house was studied in the MalariaSphere. Carbon dioxide baited traps, placed outside human dwellings, were also tested in an African village setting. The laboratory and semi-field data were analysed by a χ2-test, the field data by GLM. In addition, CO2 concentrations produced by yeast-sugar solutions were measured over time. Results Traps baited with yeast-produced CO2 caught significantly more mosquitoes than unbaited traps (up to 34 h post mixing the ingredients and also significantly more than traps baited with industrial CO2, both in the laboratory and semi-field. Adding yeast-produced CO2 to traps baited with human odour significantly increased trap catches. In the MalariaSphere, outdoor traps baited with yeast-produced or industrial CO2 + human odour reduced house entry of mosquitoes with a human host sleeping under a bed net indoors. Anopheles gambiae s.s. was not caught during the field trials

  18. Cotton pest management practices and the selection of pyrethroid resistance in Anopheles gambiae population in Northern Benin

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    Yadouleton Anges

    2011-04-01

    Full Text Available Abstract Background Pyrethroid insecticides, carbamate and organophosphate are the classes of insecticides commonly used in agriculture for crop protection in Benin. Pyrethroids remain the only class of insecticides recommended by the WHO for impregnation of bed nets. Unfortunately, the high level of pyrethroid resistance in Anopheles gambiae s.l., threatens to undermine the success of pyrethroid treated nets. This study focuses on the investigation of agricultural practices in cotton growing areas, and their direct impact on larval populations of An. gambiae in surrounding breeding sites. Methods The protocol was based on the collection of agro-sociological data where farmers were subjected to semi-structured questionnaires based on the strategies used for crop protection. This was complemented by bioassay tests to assess the susceptibility of malaria vectors to various insecticides. Molecular analysis was performed to characterize the resistance genes and the molecular forms of An. gambiae. Insecticide residues in soil samples from breeding sites were investigated to determine major factors that can inhibit the normal growth of mosquito larvae by exposing susceptible and resistant laboratory strains. Results There is a common use by local farmers of mineral fertilizer NPK at 200 kg/ha and urea at 50 kg/hectare following insecticide treatments in both the Calendar Control Program (CCP and the Targeted Intermittent Control Program (TICP. By contrast, no chemicals are involved in Biological Program (BP where farmers use organic and natural fertilizers which include animal excreta. Susceptibility test results confirmed a high resistance to DDT. Mean mortality of An. gambiae collected from the farms practicing CCP, TICP and BP methods were 33%, 42% and 65% respectively. An. gambiae populations from areas using the CCP and TICP programs showed resistance to permethrin with mortality of 50% and 58% respectively. By contrast, bioassay test results of

  19. Repellent, irritant and toxic effects of 20 plant extracts on adults of the malaria vector Anopheles gambiae mosquito.

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    Emilie Deletre

    Full Text Available Pyrethroid insecticides induce an excito-repellent effect that reduces contact between humans and mosquitoes. Insecticide use is expected to lower the risk of pathogen transmission, particularly when impregnated on long-lasting treated bednets. When applied at low doses, pyrethroids have a toxic effect, however the development of pyrethroid resistance in several mosquito species may jeopardize these beneficial effects. The need to find additional compounds, either to kill disease-carrying mosquitoes or to prevent mosquito contact with humans, therefore arises. In laboratory conditions, the effects (i.e., repellent, irritant and toxic of 20 plant extracts, mainly essential oils, were assessed on adults of Anopheles gambiae, a primary vector of malaria. Their effects were compared to those of DEET and permethrin, used as positive controls. Most plant extracts had irritant, repellent and/or toxic effects on An. gambiae adults. The most promising extracts, i.e. those combining the three types of effects, were from Cymbopogon winterianus, Cinnamomum zeylanicum and Thymus vulgaris. The irritant, repellent and toxic effects occurred apparently independently of each other, and the behavioural response of adult An. gambiae was significantly influenced by the concentration of the plant extracts. Mechanisms underlying repellency might, therefore, differ from those underlying irritancy and toxicity. The utility of the efficient plant extracts for vector control as an alternative to pyrethroids may thus be envisaged.

  20. Computational approach for identification of Anopheles gambiae miRNA involved in modulation of host immune response.

    Science.gov (United States)

    Thirugnanasambantham, Krishnaraj; Hairul-Islam, Villianur Ibrahim; Saravanan, Subramanian; Subasri, Subramaniyan; Subastri, Ariraman

    2013-05-01

    MicroRNAs (miRNAs) are small, noncoding RNAs that play key roles in regulating gene expression in animals, plants, and viruses, which involves in biological processes including development, cancer, immunity, and host-microorganism interactions. In this present study, we have used the computational approach to identify potent miRNAs involved in Anopheles gambiae immune response. Analysis of 217,261 A. gambiae ESTs and further study of RNA folding revealed six new miRNAs. The minimum free energy of the predicted miRNAs ranged from -27.2 to -62.63 kcal/mol with an average of -49.38 kcal/mol. While its A + U % ranges from 50 to 65 % with an average value of 57.37 %. Phylogenetic analysis of the predicted miRNAs revealed that aga-miR-277 was evolutionary highly conserved with more similarity with other mosquito species. Observing further the target identification of the predicted miRNA, it was noticed that the aga-miR-2304 and aga-miR-2390 are involved in modulation of immune response by targeting the gene encoding suppressin and protein prophenoloxidase. Further detailed studies of these miRNAs will help in revealing its function in modulation of A. gambiae immune response with respect to its parasite.

  1. Relationship between kdr mutation and resistance to pyrethroid and DDT insecticides in natural populations of Anopheles gambiae.

    Science.gov (United States)

    Reimer, Lisa; Fondjo, Etienne; Patchoké, Salomon; Diallo, Brehima; Lee, Yoosook; Ng, Arash; Ndjemai, Hamadou M; Atangana, Jean; Traore, Sekou F; Lanzaro, Gregory; Cornel, Anthony J

    2008-03-01

    The spread of insecticide resistance genes in Anopheles gambiae Giles sensu stricto threatens to compromise vector-based malaria control programs. Two mutations at the same locus in the voltage-gated sodium channel gene are known to confer knockdown resistance (kdr) to pyrethroids and DDT. Kdr-e involves a leucine-serine substitution, and it was until recently thought to be restricted to East Africa, whereas kdr-w, which involves a leucine-phenylalanine substitution, is associated with resistance in West Africa. In this study, we analyze the frequency and relationship between the kdr genotypes and resistance to type I and type II pyrethroids and DDT by using WHO test kits in both the Forest-M and S molecular forms of An. gambiae in Cameroon. Both kdr-w and kdr-e polymorphisms were found in sympatric An. gambiae, and in many cases in the same mosquito. Kdr-e and kdr-w were detected in both forms, but they were predominant in the S form. Both kdr-e and kdr-w were closely associated with resistance to DDT and weakly associated with resistance to type II pyrethroids. Kdr-w conferred greater resistance to permethrin than kdr-e. We also describe a modified diagnostic designed to detect both resistant alleles simultaneously.

  2. Behavioural determinants of gene flow in malaria vector populations: Anopheles gambiae males select large females as mates

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    Yan G

    2002-08-01

    Full Text Available Abstract Background Plasmodium-refractory mosquitoes are being rapidly developed for malaria control but will only succeed if they can successfully compete for mates when released into the wild. Pre-copulatory behavioural traits maintain genetic population structure in wild mosquito populations and mating barriers have foiled previous attempts to control malaria vectors through sterile male release. Methods Varying numbers of virgin male and female Anopheles gambiae Giles, from two strains of different innate sizes, were allowed to mate under standardized conditions in laboratory cages, following which, the insemination status, oviposition success and egg batch size of each female was assessed. The influence of male and female numbers, strain combination and female size were determined using logistic regression, correlation analysis and a simple mechanistic model of male competition for females. Results Male An. gambiae select females on the basis of size because of much greater fecundity among large females. Even under conditions where large numbers of males must compete for a smaller number of females, the largest females are more likely to become inseminated, to successfully oviposit and to produce large egg batches. Conclusions Sexual selection, on the basis of size, could either promote or limit the spread of malaria-refractory genes into wild populations and needs to be considered in the continued development and eventual release of transgenic vectors. Fundamental studies of behavioural ecology in malaria vectors such as An. gambiae can have important implications for malaria control and should be prioritised for more extensive investigation in the future.

  3. Molecular evolution of a gene cluster of serine proteases expressed in the Anopheles gambiae female reproductive tract

    Directory of Open Access Journals (Sweden)

    Tramontano Anna

    2011-03-01

    Full Text Available Abstract Background Genes involved in post-mating processes of multiple mating organisms are known to evolve rapidly due to coevolution driven by sexual conflict among male-female interacting proteins. In the malaria mosquito Anopheles gambiae - a monandrous species in which sexual conflict is expected to be absent or minimal - recent data strongly suggest that proteolytic enzymes specifically expressed in the female lower reproductive tissues are involved in the processing of male products transferred to females during mating. In order to better understand the role of selective forces underlying the evolution of proteins involved in post-mating responses, we analysed a cluster of genes encoding for three serine proteases that are down-regulated after mating, two of which specifically expressed in the atrium and one in the spermatheca of A. gambiae females. Results The analysis of polymorphisms and divergence of these female-expressed proteases in closely related species of the A. gambiae complex revealed a high level of replacement polymorphisms consistent with relaxed evolutionary constraints of duplicated genes, allowing to rapidly fix novel replacements to perform new or more specific functions. Adaptive evolution was detected in several codons of the 3 genes and hints of episodic selection were also found. In addition, the structural modelling of these proteases highlighted some important differences in their substrate specificity, and provided evidence that a number of sites evolving under selective pressures lie relatively close to the catalytic triad and/or on the edge of the specificity pocket, known to be involved in substrate recognition or binding. The observed patterns suggest that these proteases may interact with factors transferred by males during mating (e.g. substrates, inhibitors or pathogens and that they may have differently evolved in independent A. gambiae lineages. Conclusions Our results - also examined in light of

  4. Some strains of Plasmodium falciparum, a human malaria parasite, evade the complement-like system of Anopheles gambiae mosquitoes.

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    Molina-Cruz, Alvaro; DeJong, Randall J; Ortega, Corrie; Haile, Ashley; Abban, Ekua; Rodrigues, Janneth; Jaramillo-Gutierrez, Giovanna; Barillas-Mury, Carolina

    2012-07-10

    Plasmodium falciparum lines differ in their ability to infect mosquitoes. The Anopheles gambiae L3-5 refractory (R) line melanizes most Plasmodium species, including the Brazilian P. falciparum 7G8 line, but it is highly susceptible to some African P. falciparum strains such as 3D7, NF54, and GB4. We investigated whether these lines differ in their ability to evade the mosquito immune system. Silencing key components of the mosquito complement-like system [thioester-containing protein 1 (TEP1), leucine-rich repeat protein 1, and Anopheles Plasmodium-responsive leucine-rich repeat protein 1] prevented melanization of 7G8 parasites, reverting the refractory phenotype. In contrast, it had no effect on the intensity of infection with NF54, suggesting that this line is able to evade TEP1-mediated lysis. When R females were coinfected with a line that is melanized (7G8) and a line that survives (3D7), the coinfection resulted in mixed infections with both live and encapsulated parasites on individual midguts. This finding shows that survival of individual parasites is parasite-specific and not systemic in nature, because parasites can evade TEP1-mediated lysis even when other parasites are melanized in the same midgut. When females from an extensive genetic cross between R and susceptible A. gambiae (G3) mosquitoes were infected with P. berghei, encapsulation was strongly correlated with the TEP1-R1 allele. However, P. falciparum 7G8 parasites were no longer encapsulated by females from this cross, indicating that the TEP1-R1 allele is not sufficient to melanize this line. Evasion of the A. gambiae immune system by P. falciparum may be the result of parasite adaptation to sympatric mosquito vectors and may be an important factor driving malaria transmission. PMID:22623529

  5. Caractérisation moléculaire des moustiques du complexe Anopheles gambiae à Mayotte et à Grande Comore

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    Léong Pock Tsy J.M.

    2003-09-01

    Full Text Available Les moustiques du complexe Anopheles gambiae ont été caractérisés sur le plan spécifique et sub-spécifique dans deux îles de l'archipel des Comores : l'Ile de Mayotte (collectivité départementale française et l'Ile de Grande Comore (Union des Comores. Les résultats sont semblables pour les deux îles et sont présentés groupés. Seule l'espèce An. gambiae s.s. a été observée (détermination sur 149 spécimens par PCR sur un amplicon d'IGS de l'ADNr. Seule la forme moléculaire S, assimilable dans cette zone géographique à la forme chromosomique Savane, a été observée (détermination sur 123 spécimens par PCR sur un autre amplicon de l'IGS de l'ADNr. Enfin, seul le sous-type IB, rencontré en Afrique de l'Est, a été trouvé (détermination sur dix spécimens, par séquençage d'une zone amplifiée de l'ITS de l'ADNr, et observation de la position 871. En conclusion, à Mayotte et à Grande Comore, le complexe An. gambiae comprend uniquement des An. gambiae s.s. de la forme moléculaire S/type IB.

  6. Behavioural responses of Anopheles gambiae sensu stricto M and S molecular form larvae to an aquatic predator in Burkina Faso

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    Gimonneau Geoffrey

    2012-03-01

    Full Text Available Abstract Background Predation of aquatic immature stages has been identified as a major evolutionary force driving habitat segregation and niche partitioning in the malaria mosquito Anopheles gambiae sensu stricto in the humid savannahs of Burkina Faso, West Africa. Here, we explored behavioural responses to the presence of a predator in wild populations of the M and S molecular forms of An. gambiae that typically breed in permanent (e.g., rice field paddies and temporary (e.g., road ruts water collections. Methods Larvae used in these experiments were obtained from eggs laid by wild female An. gambiae collected from two localities in south-western Burkina Faso during the 2008 rainy season. Single larvae were observed in an experimental arena, and behavioural traits were recorded and quantified a in the absence of a predator and b in the presence of a widespread mosquito predator, the backswimmer Anisops jaczewskii. Differences in the proportion of time allocated to each behaviour were assessed using Principal Component Analysis and Multivariate Analysis of Variance. Results The behaviour of M and S form larvae was found to differ significantly; although both forms mainly foraged at the water surface, spending 60-90% of their time filtering water at the surface or along the wall of the container, M form larvae spent on average significantly more time browsing at the bottom of the container than S form larvae (4.5 vs. 1.3% of their overall time, respectively; P P P P Conclusions Behavioural differences between larvae of the M and S form of An. gambiae in response to an acute predation risk is likely to be a reflection of different trade-offs between foraging and predator vigilance that might be of adaptive value in contrasting aquatic ecosystems. Future studies should explore the relevance of these findings under the wide range of natural settings where both forms co-exist in Africa.

  7. Dynamics of knockdown pyrethroid insecticide resistance alleles in a field population of Anopheles gambiae s.s. in southwestern Nigeria

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    T.S. Awolola, A.O. Oduola, I.O. Oyewole, J.B. Obansa, C.N. Amajoh

    2007-09-01

    Full Text Available Background & objectives: Pyrethroid insecticide resistance in the malaria vector Anopheles gambiaeGiles is mainly associated with reduced target site sensitivity arising from a single point mutation inthe sodium channel gene, often referred to as knockdown resistance (kdr. This resistance mechanismis widespread in West Africa and was reported for the first time in Nigeria in 2002. Here we presentchanges in the susceptibility/resistance status of the molecular ‘M’ and ‘S’ forms of An. gambiae andthe frequency of the kdr alleles from 2002–05.Methods: Adult anophelines were sampled quarterly inside human dwellings from January 2002 toDecember 2005 and adults reared from wild larvae were identified using morphological keys. Samplesbelonging to the An. gambiae complex were subjected to PCR assays for species identification anddetection of molecular ‘M’ and ‘S’ forms. Insecticide susceptibility tests were carried out usingstandard WHO procedures and test kits only on 2–3 days old adult An. gambiae s.s. reared fromlarval collections. The kdr genotypes were determined in both live and dead specimens of An. gambiaes.s. using alleles-specific polymerase chain reaction diagnostic tests.Results: The overall collection showed that the molecular ‘S’ form was predominant (>60% but theproportions of both forms in the mosquito populations from 2002–05 were not statistically different.Both forms also occurred throughout the period without apparent relationship to wet or dry season.Insecticide susceptibility tests did not show any significant increase in the resistance status recordedfor either Permethrin or DDT from 2002–05, rather, an improvement in the susceptibility status ofthe mosquitoes to these insecticides was observed from 2004–05 relative to the tests performed in2002–03.Conclusion: The proportion of the molecular ‘M’ and ‘S’ form of An. gambiae and the kdr frequencieshave not increased significantly from 2002

  8. Co-occurrence and distribution of East (L1014S) and West (L1014F) African knock-down resistance in Anopheles gambiae sensu lato population of Tanzania

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    Kabula, Bilali; Kisinza, William; Tungu, Patrick; Ndege, Chacha; Batengana, Benard; Kollo, Douglas; Malima, Robert; Kafuko, Jessica; Mohamed, Mahdi; Magesa, Stephen

    2014-01-01

    Objective Insecticide resistance molecular markers can provide sensitive indicators of resistance development in Anopheles vector populations. Assaying these makers is of paramount importance in the resistance monitoring programme. We investigated the presence and distribution of knock-down resistance (kdr) mutations in Anopheles gambiae s.l. in Tanzania. Methods Indoor-resting Anopheles mosquitoes were collected from 10 sites and tested for insecticide resistance using the standard WHO protocol. Polymerase chain reaction-based molecular diagnostics were used to genotype mosquitoes and detect kdr mutations. Results The An. gambiae tested were resistance to lambdacyhalothrin in Muheza, Arumeru and Muleba. Out of 350 An. gambiae s.l. genotyped, 35% were An. gambiae s.s. and 65% An. arabiensis. L1014S and L1014F mutations were detected in both An. gambiae s.s. and An. arabiensis. L1014S point mutation was found at the allelic frequency of 4–33%, while L1014F was at the allelic frequency 6–41%. The L1014S mutation was much associated with An. gambiae s.s. (χ2 = 23.41; P lambdacialotrina en Muheza, Arumeru y Muleba. De 350 An. gambiae s.l. genotipados, 35% eran An. gambiae s.s. y 65% eran An. arabiensis. Se detectaron mutaciones L1014S y L1014F tanto en An. gambiae s.s. como en An. arabiensis. La mutación puntual L1014S se encontró con una frecuencia alélica de 4-33%, mientras que L1014F tenía una frecuencia alélica de 6-14%. La mutación L1014S estaba ampliamente asociada a An. gambiae s.s. (Chi-Cuadrado = 23.41; P lambdacialotrina (P < 0.001). Conclusión La simultaneidad de mutaciones de L1014S y L1014F junto con informes de resistencia a los insecticidas sugiere que la resistencia a piretroides se está convirtiendo en un fenómeno común entre las poblaciones del vector de la malaria en Tanzania. La presencia de la mutación L1014F en estas poblaciones del Este de África indican la diseminación del gen a lo largo del continente africano

  9. Alstonia boonei De Wild oil extract in the management of mosquito (Anopheles gambiae, a vector of malaria disease

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    Kayode David Ileke

    2015-07-01

    Full Text Available Objective: To evaluate the insecticidal potential of Alstonia boonei (A. boonei oils and derivatives against different life stages of a malaria vector, Anopheles gambiae. Methods: The leaf, stem bark and root bark of A. boonei were collected from an open field and air dried before being blended to fine powder. Oils from this plant were extracted by cold extraction and were prepared at different concentrations. Contact toxicity of A. boonei was tested against the larvae and pupae of the insect while smoke toxicity of the plant materials in form of mosquito coil was tested against the adult insect. Results: Alstodine recorded the highest insect mortality rate and the order of susceptibility of the life stages of the insect to the plant was pupae alstonine > stem bark extract > leaf extract > root bark extract.

  10. Alstonia booneiDe Wildoil extract in the management of mosquito (Anopheles gambiae), a vector of malaria disease

    Institute of Scientific and Technical Information of China (English)

    Kayode David Ileke; Olaniyi Charles Ogungbite

    2015-01-01

    Objective:To evaluate the insecticidal potential ofAlstonia boonei(A. boonei)oils and derivatives against different life stages of a malaria vector,Anopheles gambiae. Methods:The leaf, stem bark and root bark ofA. boonei were collected from an open field and air dried before being blended to fine powder. Oils from this plant were extracted by cold extraction and were prepared at different concentrations. Contact toxicity ofA. boonei was tested against the larvae and pupae of the insect while smoke toxicity of the plant materials in form of mosquito coil was tested against the adult insect. Results: Alstodine recorded the highest insect mortality rate and the order of susceptibility of the life stages of the insect to the plant was pupae alstonine > stem bark extract > leaf extract > root bark extract.

  11. Salivary gland proteome analysis reveals modulation of anopheline unique proteins in insensitive acetylcholinesterase resistant Anopheles gambiae mosquitoes.

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    Sylvie Cornelie

    Full Text Available Insensitive acetylcholinesterase resistance due to a mutation in the acetylcholinesterase (ace encoding ace-1 gene confers cross-resistance to organophosphate and carbamate insecticides in Anopheles gambiae populations from Central and West Africa. This mutation is associated with a strong genetic cost revealed through alterations of some life history traits but little is known about the physiological and behavioural changes in insects bearing the ace-1(R allele. Comparative analysis of the salivary gland contents between An. gambiae susceptible and ace-1(R resistant strains was carried out to charaterize factors that could be involved in modifications of blood meal process, trophic behaviour or pathogen interaction in the insecticide-resistant mosquitoes. Differential analysis of the salivary gland protein profiles revealed differences in abundance for several proteins, two of them showing major differences between the two strains. These two proteins identified as saglin and TRIO are salivary gland-1 related proteins, a family unique to anopheline mosquitoes, one of them playing a crucial role in salivary gland invasion by Plasmodium falciparum sporozoites. Differential expression of two other proteins previously identified in the Anopheles sialome was also observed. The differentially regulated proteins are involved in pathogen invasion, blood feeding process, and protection against oxidation, relevant steps in the outcome of malaria infection. Further functional studies and insect behaviour experiments would confirm the impact of the modification of the sialome composition on blood feeding and pathogen transmission abilities of the resistant mosquitoes. The data supports the hypothesis of alterations linked to insecticide resistance in the biology of the primary vector of human malaria in Africa.

  12. Change in composition of the Anopheles gambiae complex and its possible implications for the transmission of malaria and lymphatic filariasis in north-eastern Tanzania

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    Derua Yahya A

    2012-06-01

    Full Text Available Abstract Background A dramatic decline in the incidence of malaria due to Plasmodium falciparum infection in coastal East Africa has recently been reported to be paralleled (or even preceded by an equally dramatic decline in malaria vector density, despite absence of organized vector control. As part of investigations into possible causes for the change in vector population density, the present study analysed the Anopheles gambiae s.l. sibling species composition in north-eastern Tanzania. Methods The study was in two parts. The first compared current species complex composition in freshly caught An. gambiae s.l. complex from three villages to the composition reported from previous studies carried out 2–4 decades ago in the same villages. The second took advantage of a sample of archived dried An. gambiae s.l. complex specimens collected regularly from a fourth study village since 2005. Both fresh and archived dried specimens were identified to sibling species of the An. gambiae s.l. complex by PCR. The same specimens were moreover examined for Plasmodium falciparum and Wuchereria bancrofti infection by PCR. Results As in earlier studies, An. gambiae s.s., Anopheles merus and Anopheles arabiensis were identified as sibling species found in the area. However, both study parts indicated a marked change in sibling species composition over time. From being by far the most abundant in the past An. gambiae s.s. was now the most rare, whereas An. arabiensis had changed from being the most rare to the most common. P. falciparum infection was rarely detected in the examined specimens (and only in An. arabiensis whereas W. bancrofti infection was prevalent and detected in all three sibling species. Conclusion The study indicates that a major shift in An. gambiae s.l. sibling species composition has taken place in the study area in recent years. Combined with the earlier reported decline in overall malaria vector density, the study suggests that this

  13. The Anopheles gambiae oxidation resistance 1 (OXR1 gene regulates expression of enzymes that detoxify reactive oxygen species.

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    Giovanna Jaramillo-Gutierrez

    Full Text Available BACKGROUND: OXR1 is an ancient gene, present in all eukaryotes examined so far that confers protection from oxidative stress by an unknown mechanism. The most highly conserved region of the gene is the carboxyl-terminal TLDc domain, which has been shown to be sufficient to prevent oxidative damage. METHODOLOGY/PRINCIPAL FINDINGS: OXR1 has a complex genomic structure in the mosquito A. gambiae, and we confirm that multiple splice forms are expressed in adult females. Our studies revealed that OXR1 regulates the basal levels of catalase (CAT and glutathione peroxidase (Gpx expression, two enzymes involved in detoxification of hydrogen peroxide, giving new insight into the mechanism of action of OXR1. Gene silencing experiments indicate that the Jun Kinase (JNK gene acts upstream of OXR1 and also regulates expression of CAT and GPx. Both OXR1 and JNK genes are required for adult female mosquitoes to survive chronic oxidative stress. OXR1 silencing decreases P. berghei oocyst formation. Unexpectedly, JNK silencing has the opposite effect and enhances Plasmodium infection in the mosquito, suggesting that JNK may also mediate some, yet to be defined, antiparasitic response. CONCLUSION: The JNK pathway regulates OXR1 expression and OXR1, in turn, regulates expression of enzymes that detoxify reactive oxygen species (ROS in Anopheles gambiae. OXR1 silencing decreases Plasmodium infection in the mosquito, while JNK silencing has the opposite effect and enhances infection.

  14. Reactive oxygen species detoxification by catalase is a major determinant of fecundity in the mosquito Anopheles gambiae.

    Science.gov (United States)

    DeJong, Randall J; Miller, Lisa M; Molina-Cruz, Alvaro; Gupta, Lalita; Kumar, Sanjeev; Barillas-Mury, Carolina

    2007-02-13

    The mosquito Anopheles gambiae is a primary vector of Plasmodium parasites in Africa. The effect of aging on reproductive output in A. gambiae females from three strains that differ in their ability to melanize Plasmodium and in their systemic levels of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), was analyzed. The number of eggs oviposited after the first blood meal decreases with age in all strains; however, this decline was much more pronounced in the G3 (unselected) and R (refractory to Plasmodium infection) strains than in the S (highly susceptible to Plasmodium) strain. Reduction of ROS levels in G3 and R females by administration of antioxidants reversed this age-related decline in fecundity. The S and G3 strains were fixed for two functionally different catalase alleles that differ at the second amino acid position (Ser2Trp). Biochemical analysis of recombinant proteins revealed that the Trp isoform has lower specific activity and higher Km than the Ser isoform, indicating that the former is a less efficient enzyme. The Trp-for-Ser substitution appears to destabilize the functional tetrameric form of the enzyme. Both alleles are present in the R strain, and Ser/Ser females had significantly higher fecundity than Trp/Trp females. Finally, a systemic reduction in catalase activity by dsRNA-mediated knockdown significantly reduced the reproductive output of mosquito females, indicating that catalase plays a central role in protecting the oocyte and early embryo from ROS damage. PMID:17284604

  15. The Anopheles gambiae Oxidation Resistance 1 (OXR1) Gene Regulates Expression of Enzymes That Detoxify Reactive Oxygen Species

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    Jaramillo-Gutierrez, Giovanna; Molina-Cruz, Alvaro; Kumar, Sanjeev; Barillas-Mury, Carolina

    2010-01-01

    Background OXR1 is an ancient gene, present in all eukaryotes examined so far that confers protection from oxidative stress by an unknown mechanism. The most highly conserved region of the gene is the carboxyl-terminal TLDc domain, which has been shown to be sufficient to prevent oxidative damage. Methodology/Principal Findings OXR1 has a complex genomic structure in the mosquito A. gambiae, and we confirm that multiple splice forms are expressed in adult females. Our studies revealed that OXR1 regulates the basal levels of catalase (CAT) and glutathione peroxidase (Gpx) expression, two enzymes involved in detoxification of hydrogen peroxide, giving new insight into the mechanism of action of OXR1. Gene silencing experiments indicate that the Jun Kinase (JNK) gene acts upstream of OXR1 and also regulates expression of CAT and GPx. Both OXR1 and JNK genes are required for adult female mosquitoes to survive chronic oxidative stress. OXR1 silencing decreases P. berghei oocyst formation. Unexpectedly, JNK silencing has the opposite effect and enhances Plasmodium infection in the mosquito, suggesting that JNK may also mediate some, yet to be defined, antiparasitic response. Conclusion The JNK pathway regulates OXR1 expression and OXR1, in turn, regulates expression of enzymes that detoxify reactive oxygen species (ROS) in Anopheles gambiae. OXR1 silencing decreases Plasmodium infection in the mosquito, while JNK silencing has the opposite effect and enhances infection. PMID:20567517

  16. Reactive oxygen species detoxification by catalase is a major determinant of fecundity in the mosquito Anopheles gambiae

    Science.gov (United States)

    DeJong, Randall J.; Miller, Lisa M.; Molina-Cruz, Alvaro; Gupta, Lalita; Kumar, Sanjeev; Barillas-Mury, Carolina

    2007-01-01

    The mosquito Anopheles gambiae is a primary vector of Plasmodium parasites in Africa. The effect of aging on reproductive output in A. gambiae females from three strains that differ in their ability to melanize Plasmodium and in their systemic levels of hydrogen peroxide (H2O2), a reactive oxygen species (ROS), was analyzed. The number of eggs oviposited after the first blood meal decreases with age in all strains; however, this decline was much more pronounced in the G3 (unselected) and R (refractory to Plasmodium infection) strains than in the S (highly susceptible to Plasmodium) strain. Reduction of ROS levels in G3 and R females by administration of antioxidants reversed this age-related decline in fecundity. The S and G3 strains were fixed for two functionally different catalase alleles that differ at the second amino acid position (Ser2Trp). Biochemical analysis of recombinant proteins revealed that the Trp isoform has lower specific activity and higher Km than the Ser isoform, indicating that the former is a less efficient enzyme. The Trp-for-Ser substitution appears to destabilize the functional tetrameric form of the enzyme. Both alleles are present in the R strain, and Ser/Ser females had significantly higher fecundity than Trp/Trp females. Finally, a systemic reduction in catalase activity by dsRNA-mediated knockdown significantly reduced the reproductive output of mosquito females, indicating that catalase plays a central role in protecting the oocyte and early embryo from ROS damage. PMID:17284604

  17. Insecticidal Activities of Bark, Leaf and Seed Extracts of Zanthoxylum heitzii against the African Malaria Vector Anopheles gambiae

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    Hans J. Overgaard

    2014-12-01

    Full Text Available The olon tree, Zanthoxylum heitzii (syn. Fagara heitzii is commonly found in the central-west African forests. In the Republic of Congo (Congo-Brazzaville its bark is anecdotally reported to provide human protection against fleas. Here we assess the insecticidal activities of Z. heitzii stem bark, seed and leaf extracts against Anopheles gambiae s.s, the main malaria vector in Africa. Extracts were obtained by Accelerated Solvent Extraction (ASE using solvents of different polarity and by classical Soxhlet extraction using hexane as solvent. The insecticidal effects of the crude extracts were evaluated using topical applications of insecticides on mosquitoes of a susceptible reference strain (Kisumu [Kis], a strain homozygous for the L1014F kdr mutation (kdrKis, and a strain homozygous for the G119S Ace1R allele (AcerKis. The insecticidal activities were measured using LD50 and LD95 and active extracts were characterized by NMR spectroscopy and HPLC chromatography. Results show that the ASE hexane stem bark extract was the most effective compound against An. gambiae (LD50 = 102 ng/mg female, but was not as effective as common synthetic insecticides. Overall, there was no significant difference between the responses of the three mosquito strains to Z. heitzii extracts, indicating no cross resistance with conventional pesticides.

  18. Acetylcholinesterases from the Disease Vectors Aedes aegypti and Anopheles gambiae: Functional Characterization and Comparisons with Vertebrate Orthologues.

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    Cecilia Engdahl

    Full Text Available Mosquitoes of the Anopheles (An. and Aedes (Ae. genus are principal vectors of human diseases including malaria, dengue and yellow fever. Insecticide-based vector control is an established and important way of preventing transmission of such infections. Currently used insecticides can efficiently control mosquito populations, but there are growing concerns about emerging resistance, off-target toxicity and their ability to alter ecosystems. A potential target for the development of insecticides with reduced off-target toxicity is the cholinergic enzyme acetylcholinesterase (AChE. Herein, we report cloning, baculoviral expression and functional characterization of the wild-type AChE genes (ace-1 from An. gambiae and Ae. aegypti, including a naturally occurring insecticide-resistant (G119S mutant of An. gambiae. Using enzymatic digestion and liquid chromatography-tandem mass spectrometry we found that the secreted proteins were post-translationally modified. The Michaelis-Menten constants and turnover numbers of the mosquito enzymes were lower than those of the orthologous AChEs from Mus musculus and Homo sapiens. We also found that the G119S substitution reduced the turnover rate of substrates and the potency of selected covalent inhibitors. Furthermore, non-covalent inhibitors were less sensitive to the G119S substitution and differentiate the mosquito enzymes from corresponding vertebrate enzymes. Our findings indicate that it may be possible to develop selective non-covalent inhibitors that effectively target both the wild-type and insecticide resistant mutants of mosquito AChE.

  19. Effects of adult body size on fecundity and the pre-gravid rate of Anopheles gambiae females in Tanzania.

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    Lyimo, E O; Takken, W

    1993-10-01

    The influence of adult body size on the pre-gravid state and fecundity was studied in Anopheles gambiae Giles females hand-caught inside houses and virgin females collected as pupae in Tanzania. Blood-fed mosquitoes were kept for 2-3 days before dissection and examination for insemination and ovarian condition. Those females which did not develop eggs were classified as pre-gravid. The number of mature eggs in those mosquitoes which became gravid was counted. Virgin females were fed and kept for egg maturation in the laboratory. Wing-length of females was measured as an index of mosquito size. The overall pre-gravid rate in the resting An.gambiae population was found to be 21% and, of these, 66% had been inseminated. In the virgin females the pre-gravid rate was 92.6%. The mean wing-length of wild females which became gravid was significantly larger than those which remained pre-gravid. There was a positive correlation between fecundity and wing-length. Smaller females tended to require two or three bloodmeals to facilitate completion of the first gonotrophic cycle. The critical size permitting oviposition from the first blood-meal was a wing-length of 3 mm. PMID:8268486

  20. Observation of granulations in the basal body of ovarioles and follicular dilatations for the determination of physiological age ofAnopheles gambiaes.s.

    Institute of Scientific and Technical Information of China (English)

    Rodrigue Anagonou; Virgile Gnanguenon; Fiacre Agossa; Bruno Akinro; Armand Akpo; Martial Gbegbo; Albert Salako; Martin Akogbto

    2015-01-01

    Objective:To explore ovariole basal body granulations and follicular dilatations for determining physiological age inAnopheles gambiaes.s.(An. gambiaes.s.). Methods: Mosquitoes were collected by using window trap catch and identified morphologically. For the first lot ofmosquitoes, they were dissected, and ovary was left in distilled water for reading ovarian tracheoles and the second was cut and transferred to another blade in a physiological liquid for verification of ovariole basal body granulations. The same approach was followed with the second lot of mosquitoes where follicular dilatations were found after classic dilaceration of ovaries were transferred into physiological liquid. The other body parts of mosquitoes were used to identify the species of theAn. gambiaes.s. complex by PCR. Results:Among the 123An. gambiae s.s. of the first lot, the method of Detinova determined the age of 89 mosquitoes versus 114 for the observation of granulations (P > 0.05). Among the 112An. gambiae s.s. of the second lot, the method of Detinova determined the age of 84 mosquitoes versus 93 for the observation of follicular dilatations (P > 0.05). Unlike the method of Detinova, observation of follicular dilatations and basal body granulations of ovarioles were possible beyond the stage II Christophers. Conclusions: Overall, the observation of follicular dilatations and ovariole basal body granulations are reliable for the determination of the physiological age inAn. gambiaes.s. Furthermore, these two methods can be used beyond the stage II.

  1. Staggered larval time-to-hatch and insecticide resistance in the major malaria vector Anopheles gambiae S form

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    Coetzee Maureen

    2010-12-01

    Full Text Available Abstract Background Anopheles gambiae is a major vector of malaria in the West African region. Resistance to multiple insecticides has been recorded in An. gambiae S form in the Ahafo region of Ghana. A laboratory population (GAH established using wild material from this locality has enabled a mechanistic characterization of each resistance phenotype as well as an analysis of another adaptive characteristic - staggered larval time-to-hatch. Methods Individual egg batches obtained from wild caught females collected from Ghana and the Republic of the Congo were monitored for staggered larval time-to-hatch. In addition, early and late larval time-to-hatch sub-colonies were selected from GAH. These selected sub-colonies were cross-mated and their hybrid progeny were subsequently intercrossed and back-crossed to the parental strains. The insecticide susceptibilities of the GAH base colony and the time-to-hatch selected sub-colonies were quantified for four insecticide classes using insecticide bioassays. Resistance phenotypes were mechanistically characterized using insecticide-synergist bioassays and diagnostic molecular assays for known reduced target-site sensitivity mutations. Results Anopheles gambiae GAH showed varying levels of resistance to all insecticide classes. Metabolic detoxification and reduced target-site sensitivity mechanisms were implicated. Most wild-caught families showed staggered larval time-to-hatch. However, some families were either exclusively early hatching or late hatching. Most GAH larvae hatched early but many egg batches contained a proportion of late hatching larvae. Crosses between the time-to-hatch selected sub-colonies yielded ambiguous results that did not fit any hypothetical models based on single-locus Mendelian inheritance. There was significant variation in the expression of insecticide resistance between the time-to-hatch phenotypes. Conclusions An adaptive response to the presence of multiple insecticide

  2. [Anopheles gambiae, major malaria vector in Logbessou, a peri-urban area of Douala (Cameroon)].

    Science.gov (United States)

    Akono, P Ntonga; Tonga, C; Mbida, J A Mbida; Hondt, O E Ngo; Ambene, P Awono; Ndo, C; Magne, G Tamdem; Peka, M F; Ngaha, R; Lehman, L G

    2015-12-01

    An entomological survey was carried out from August to November 2013, in order to determine the vector system of a building site for social housing in a coastal periurban district of Douala (Cameroon). Mosquito larvae were collected and adult endophilic mosquitoes captured on volunteers, for a total sample of 4897 mosquitoes. Morpho-taxonomic techniques alongside molecular techniques enabled the identification of 4 species, all aggressive to humans: Cx. pipiens (22.3%), Ae. albopictus (0.3%), An. coluzzii and An. gambiae (77.4%). The overall average biting rate recorded was 41.73 bites/person/night (b/p/n). An. gambiae s.l. represents 90.82% of this aggressive fauna, followed by Cx. pipiens (8.58%) and Ae. albopictus (0.6%). The detection of CSP showed that An. gambiae was responsible for 100% of P. falciparum transmission. The overall mean Entomological Inoculation Rate (EIR) was 3.94 ib/p/n. Female An. gambiae mortality rates were 14.47%, 82.5% and 100% respectively with DDT, permethrin and deltamethrin. The proliferation of An. gambiae in this area during raining season, at the detriment of An. coluzzii Coetze & Wilkerson and An. melas Theobald known to be major malaria vectors in island and coastal areas of Africa, may owe to the forest that still colonises this coastal peri-urban locality. Residents should therefore make use of deltamethrin based protective measures.

  3. Dynamics of insecticide resistance in malaria vectors in Benin: first evidence of the presence of L1014S kdr mutation in Anopheles gambiae from West Africa

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    Ranson Hilary

    2011-09-01

    Full Text Available Abstract Background Insecticide resistance monitoring is essential to help national programmers to implement more effective and sustainable malaria control strategies in endemic countries. This study reported the spatial and seasonal variations of insecticide resistance in malaria vectors in Benin, West Africa. Methods Anopheles gambiae s.l populations were collected from October 2008 to June 2010 in four sites selected on the basis of different use of insecticides and environment. WHO susceptibility tests were carried out to detect resistance to DDT, fenitrothion, bendiocarb, permethrin and deltamethrin. The synergist piperonyl butoxide was used to assess the role of non-target site mechanisms in pyrethroid resistance. Anopheles gambiae mosquitoes were identified to species and to molecular M and S forms using PCR techniques. Molecular and biochemical assays were carried out to determine kdr and Ace.1R allelic frequencies and activity of the detoxification enzymes. Results Throughout the surveys very high levels of mortality to bendiocarb and fenitrothion were observed in An. gambiae s.l. populations. However, high frequencies of resistance to DDT and pyrethroids were seen in both M and S form of An. gambiae s.s. and Anopheles arabiensis. PBO increased the toxicity of permethrin and restored almost full susceptibility to deltamethrin. Anopheles gambiae s.l. mosquitoes from Cotonou and Malanville showed higher oxidase activity compared to the Kisumu susceptible strain in 2009, whereas the esterase activity was higher in the mosquitoes from Bohicon in both 2008 and 2009. A high frequency of 1014F kdr allele was initially showed in An. gambiae from Cotonou and Tori-Bossito whereas it increased in mosquitoes from Bohicon and Malanville during the second year. For the first time the L1014S kdr mutation was found in An. arabiensis in Benin. The ace.1R mutation was almost absent in An. gambiae s.l. Conclusion Pyrethroid and DDT resistance is

  4. Evidence of carbamate resistance in urban populations of Anopheles gambiae s.s. mosquitoes resistant to DDT and deltamethrin insecticides in Lagos, South-Western Nigeria

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    Oduola Adedayo O

    2012-06-01

    Full Text Available Abstract Background Resistance monitoring is essential in ensuring the success of insecticide based vector control programmes. This study was carried out to assess the susceptibility status of urban populations of Anopheles gambiae to carbamate insecticide being considered for vector control in mosquito populations previously reported to be resistant to DDT and permethrin. Methods Two – three day old adult female Anopheles mosquitoes reared from larval collections in 11 study sites from Local Government Areas of Lagos were exposed to test papers impregnated with DDT 4%, deltamethrin 0.05% and propoxur 0.1% insecticides. Additional tests were carried out to determine the susceptibility status of the Anopheles gambiae population to bendiocarb insecticide. Members of the A. gambiae complex, the molecular forms, were identified by PCR assays. The involvement of metabolic enzymes in carbamate resistance was assessed using Piperonyl butoxide (PBO synergist assays. The presence of kdr-w/e and ace-1R point mutations responsible for DDT-pyrethroid and carbamate resistance mechanisms was also investigated by PCR. Results Propoxur resistance was found in 10 out of the 11 study sites. Resistance to three classes of insecticides was observed in five urban localities. Mortality rates in mosquitoes exposed to deltamethrin and propoxur did not show any significant difference (P > 0.05 but was significantly higher (P A. gambiae s.s (M form. The kdr -w point mutation at allelic frequencies between 45%-77% was identified as one of the resistant mechanisms responsible for DDT and pyrethroid resistance. Ace-1R point mutation was absent in the carbamate resistant population. However, the possible involvement of metabolic resistance was confirmed by synergistic assays conducted. Conclusion Evidence of carbamate resistance in A. gambiae populations already harbouring resistance to DDT and permethrin is a clear indication that calls for the implementation of

  5. Development of vegetable farming: a cause of the emergence of insecticide resistance in populations of Anopheles gambiae in urban areas of Benin

    OpenAIRE

    Braïma James; Djouaka Rousseau F; Asidi Alex; Yadouleton Anges; Agossou Christian D; Akogbeto Martin C

    2009-01-01

    Abstract Background A fast development of urban agriculture has recently taken place in many areas in the Republic of Benin. This study aims to assess the rapid expansion of urban agriculture especially, its contribution to the emergence of insecticide resistance in populations of Anopheles gambiae. Methods The protocol was based on the collection of sociological data by interviewing vegetable farmers regarding various agricultural practices and the types of pesticides used. Bioassay tests we...

  6. Differential gene expression in abdomens of the malaria vector mosquito, Anopheles gambiae, after sugar feeding, blood feeding and Plasmodium berghei infection

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    Romans Patricia A; Kern Marcia K; Hillenmeyer Maureen E; Lobo Neil F; Dana Ali N; Collins Frank H

    2006-01-01

    Abstract Background Large scale sequencing of cDNA libraries can provide profiles of genes expressed in an organism under defined biological and environmental circumstances. We have analyzed sequences of 4541 Expressed Sequence Tags (ESTs) from 3 different cDNA libraries created from abdomens from Plasmodium infection-susceptible adult female Anopheles gambiae. These libraries were made from sugar fed (S), rat blood fed (RB), and P. berghei-infected (IRB) mosquitoes at 30 hours after the bloo...

  7. The complete mitochondrial genome of Anopheles minimus (Diptera: Culicidae) and the phylogenetics of known Anopheles mitogenomes.

    Science.gov (United States)

    Hua, Ya-Qiong; Ding, Yi-Ran; Yan, Zhen-Tian; Si, Feng-Ling; Luo, Qian-Chun; Chen, Bin

    2016-06-01

    Anopheles minimus is an important vector of human malaria in southern China and Southeast Asia. The phylogenetics of mosquitoes has not been well resolved, and the mitochondrial genome (mtgenome) has proven to be an important marker in the study of evolutionary biology. In this study, the complete mtgenome of An. minimus was sequenced for the first time. It is 15 395 bp long and encodes 37 genes, including 13 protein-coding genes (PCGs), 22 transfer RNAs (tRNAs), two ribosomal RNAs (rRNAs) and a non-coding region. The gene organization is consistent with those of known Anopheles mtgenomes. The mtgenome performs a clear bias in nucleotide composition with a positive AT-skew and a negative GC-skew. All 13 PCGs prefer to use the codon UUA (Leu), ATN as initiation codon but cytochrome-oxidase subunit 1 (COI) and ND5, with TCG and GTG, and TAA as termination codon, but COI, COII, COIII and ND4, all with the incomplete T. tRNAs have the typical clover-leaf structure, but tRNA(Ser(AGN)) is consistent with known Anopheles mtgenomes. The control region includes a conserved T-stretch and a (TA)n stretch, and has the highest A+T content at 93.1%. The phylogenetics of An. minimus with 18 other Anopheles species was constructed by maximum likelihood and Bayesian inference, based on concatenated PCG sequences. The subgenera, Cellia and Anopheles, and Nyssorhynchus and Kerteszia have mutually close relationships, respectively. The Punctulatus group and Leucosphyrus group of Neomyzomyia Series, and the Albitarsis group of Albitarsis Series were suggested to be monophyletic. The monophyletic status of the subgenera, Cellia, Anopheles, Nyssorhynchus and Kerteszia need to be further elucidated. PMID:26852698

  8. Inducible immune factors of the vector mosquito Anopheles gambiae: biochemical purification of a defensin antibacterial peptide and molecular cloning of preprodefensin cDNA.

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    Richman, A M; Bulet, P; Hetru, C; Barillas-Mury, C; Hoffmann, J A; Kafalos, F C

    1996-08-01

    Larvae of the mosquito vector of human malaria, Anopheles gambiae, were inoculated with bacteria and extracts were biochemically fractionated by reverse-phase HPLC. Multiple induced polypeptides and antibacterial activities were observed following bacterial infection, including a member of the insect defensin family of antibacterial proteins. A cDNA encoding An. gambiae preprodefensin was isolated using PCR primers based on phylogenetically conserved sequences. The mature peptide is highly conserved, but the signal and propeptide segments are not, relative to corresponding defensin sequences of other insects. Defensin expression is induced in response to bacterial infection, in both adult and larval stages. In contrast, pupae express defensin mRNA constitutively. Defensin expression may prove a valuable molecular marker to monitor the An. gambiae host response to infection by parasitic protozoa of medical importance. PMID:8799739

  9. Comparative evaluation of systemic drugs for their effects against Anopheles gambiae

    OpenAIRE

    Butters, Matthew P.; Kobylinski, Kevin C.; Deus, Kelsey M.; da Silva, Ines Marques; GRAY, MEG; sylla, massamba; Foy, Brian D.

    2011-01-01

    Laboratory and field studies have shown that ivermectin, a drug that targets invertebrate ligand-gated ion channels (LGICs), is potently active against Anopheles spp. mosquitoes at concentrations present in human blood after standard drug administrations; thus ivermectin holds promise as a mass human-administered endectocide that could help suppress malaria parasite transmission. We evaluated other systemic LGIC-targeting drugs for their activities against the African malaria vector Anopheles...

  10. Resistance Mechanisms of Anopheles stephensi (Diptera: Culicidae) to Temephos

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    Aboozar Soltani; Hassan Vatandoost; MohammadAli Oshaghi; Naseh Maleki-Ravasan; AhmadAli Enayati; Fatemeh Asgarian

    2014-01-01

    Background: Anopheles stephensi is a sub-tropical species and has been considered as one of the most important vector of human malaria throughout the Middle East and South Asian region including the malarious areas of southern Iran. Current reports confirmed An. stephensi resistance to temephos in Oman and India. However, there is no comprehensive research on mechanisms of temephos resistance in An. stephensi in the literature. This study was designed in order to clarify the enzymatic and mol...

  11. Apolipophorin-III mediates antiplasmodial epithelial responses in Anopheles gambiae (G3 mosquitoes.

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    Lalita Gupta

    Full Text Available BACKGROUND: Apolipophorin-III (ApoLp-III is known to play an important role in lipid transport and innate immunity in lepidopteran insects. However, there is no evidence of involvement of ApoLp-IIIs in the immune responses of dipteran insects such as Drosophila and mosquitoes. METHODOLOGY/PRINCIPAL FINDINGS: We report the molecular and functional characterization of An. gambiae apolipophorin-III (AgApoLp-III. Mosquito ApoLp-IIIs have diverged extensively from those of lepidopteran insects; however, the predicted tertiary structure of AgApoLp-III is similar to that of Manduca sexta (tobacco hornworm. We found that AgApoLp-III mRNA expression is strongly induced in the midgut of An. gambiae (G3 strain mosquitoes in response to Plasmodium berghei infection. Furthermore, immunofluorescence stainings revealed that high levels of AgApoLp-III protein accumulate in the cytoplasm of Plasmodium-invaded cells and AgApoLp-III silencing increases the intensity of P. berghei infection by five fold. CONCLUSION: There are broad differences in the midgut epithelial responses to Plasmodium invasion between An. gambiae strains. In the G3 strain of An. gambiae AgApoLp-III participates in midgut epithelial defense responses that limit Plasmodium infection.

  12. Apolipophorin-III Mediates Antiplasmodial Epithelial Responses in Anopheles gambiae (G3) Mosquitoes

    Science.gov (United States)

    Jo, Yong Hun; Oh, Seung Han; Kumar, Sanjeev; Noh, Mi Young; Lee, Yong Seok; Cha, Sung-Jae; Seo, Sook Jae; Kim, Iksoo; Han, Yeon Soo; Barillas-Mury, Carolina

    2010-01-01

    Background Apolipophorin-III (ApoLp-III) is known to play an important role in lipid transport and innate immunity in lepidopteran insects. However, there is no evidence of involvement of ApoLp-IIIs in the immune responses of dipteran insects such as Drosophila and mosquitoes. Methodology/Principal Findings We report the molecular and functional characterization of An. gambiae apolipophorin-III (AgApoLp-III). Mosquito ApoLp-IIIs have diverged extensively from those of lepidopteran insects; however, the predicted tertiary structure of AgApoLp-III is similar to that of Manduca sexta (tobacco hornworm). We found that AgApoLp-III mRNA expression is strongly induced in the midgut of An. gambiae (G3 strain) mosquitoes in response to Plasmodium berghei infection. Furthermore, immunofluorescence stainings revealed that high levels of AgApoLp-III protein accumulate in the cytoplasm of Plasmodium-invaded cells and AgApoLp-III silencing increases the intensity of P. berghei infection by five fold. Conclusion There are broad differences in the midgut epithelial responses to Plasmodium invasion between An. gambiae strains. In the G3 strain of An. gambiae AgApoLp-III participates in midgut epithelial defense responses that limit Plasmodium infection. PMID:21072214

  13. Wide cross-reactivity between Anopheles gambiae and Anopheles funestus SG6 salivary proteins supports exploitation of gSG6 as a marker of human exposure to major malaria vectors in tropical Africa

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    Petrarca Vincenzo

    2011-07-01

    Full Text Available Abstract Background The Anopheles gambiae gSG6 is an anopheline-specific salivary protein which helps female mosquitoes to efficiently feed on blood. Besides its role in haematophagy, gSG6 is immunogenic and elicits in exposed individuals an IgG response, which may be used as indicator of exposure to the main African malaria vector A. gambiae. However, malaria transmission in tropical Africa is sustained by three main vectors (A. gambiae, Anopheles arabiensis and Anopheles funestus and a general marker, reflecting exposure to at least these three species, would be especially valuable. The SG6 protein is highly conserved within the A. gambiae species complex whereas the A. funestus homologue, fSG6, is more divergent (80% identity with gSG6. The aim of this study was to evaluate cross-reactivity of human sera to gSG6 and fSG6. Methods The A. funestus SG6 protein was expressed/purified and the humoral response to gSG6, fSG6 and a combination of the two antigens was compared in a population from a malaria hyperendemic area of Burkina Faso where both vectors were present, although with a large A. gambiae prevalence (>75%. Sera collected at the beginning and at the end of the high transmission/rainy season, as well as during the following low transmission/dry season, were analysed. Results According to previous observations, both anti-SG6 IgG level and prevalence decreased during the low transmission/dry season and showed a typical age-dependent pattern. No significant difference in the response to the two antigens was found, although their combined use yielded in most cases higher IgG level. Conclusions Comparative analysis of gSG6 and fSG6 immunogenicity to humans suggests the occurrence of a wide cross-reactivity, even though the two proteins carry species-specific epitopes. This study supports the use of gSG6 as reliable indicator of exposure to the three main African malaria vectors, a marker which may be useful to monitor malaria transmission

  14. Crystal and Solution Studies of the “Plus-C” Odorant-binding Protein 48 from Anopheles gambiae

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    Tsitsanou, Katerina E.; Drakou, Christina E.; Thireou, Trias; Vitlin Gruber, Anna; Kythreoti, Georgia; Azem, Abdussalam; Fessas, Dimitrios; Eliopoulos, Elias; Iatrou, Kostas; Zographos, Spyros E.

    2013-01-01

    Much physiological and behavioral evidence has been provided suggesting that insect odorant-binding proteins (OBPs) are indispensable for odorant recognition and thus are appealing targets for structure-based discovery and design of novel host-seeking disruptors. Despite the fact that more than 60 putative OBP-encoding genes have been identified in the malaria vector Anopheles gambiae, the crystal structures of only six of them are known. It is therefore clear that OBP structure determination constitutes the bottleneck for structure-based approaches to mosquito repellent/attractant discovery. Here, we describe the three-dimensional structure of an A. gambiae “Plus-C” group OBP (AgamOBP48), which exhibits the second highest expression levels in female antennae. This structure represents the first example of a three-dimensional domain-swapped dimer in dipteran species. A combined binding site is formed at the dimer interface by equal contribution of each monomer. Structural comparisons with the monomeric AgamOBP47 revealed that the major structural difference between the two Plus-C proteins localizes in their N- and C-terminal regions, and their concerted conformational change may account for monomer-swapped dimer conversion and furthermore the formation of novel binding pockets. Using a combination of gel filtration chromatography, differential scanning calorimetry, and analytical ultracentrifugation, we demonstrate the AgamOBP48 dimerization in solution. Eventually, molecular modeling calculations were used to predict the binding mode of the most potent synthetic ligand of AgamOBP48 known so far, discovered by ligand- and structure-based virtual screening. The structure-aided identification of multiple OBP binders represents a powerful tool to be employed in the effort to control transmission of the vector-borne diseases. PMID:24097978

  15. Losing identity: structural diversity of transposable elements belonging to different classes in the genome of Anopheles gambiae

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    Fernández-Medina Rita D

    2012-06-01

    Full Text Available Abstract Background Transposable elements (TEs, both DNA transposons and retrotransposons, are genetic elements with the main characteristic of being able to mobilize and amplify their own representation within genomes, utilizing different mechanisms of transposition. An almost universal feature of TEs in eukaryotic genomes is their inability to transpose by themselves, mainly as the result of sequence degeneration (by either mutations or deletions. Most of the elements are thus either inactive or non-autonomous. Considering that the bulk of some eukaryotic genomes derive from TEs, they have been conceived as “TE graveyards.” It has been shown that once an element has been inactivated, it progressively accumulates mutations and deletions at neutral rates until completely losing its identity or being lost from the host genome; however, it has also been shown that these “neutral sequences” might serve as raw material for domestication by host genomes. Results We have analyzed the sequence structural variations, nucleotide divergence, and pattern of insertions and deletions of several superfamilies of TEs belonging to both class I (long terminal repeats [LTRs] and non-LTRs [NLTRs] and II in the genome of Anopheles gambiae, aiming at describing the landscape of deterioration of these elements in this particular genome. Our results describe a great diversity in patterns of deterioration, indicating lineage-specific differences including the presence of Solo-LTRs in the LTR lineage, 5′-deleted NLTRs, and several non-autonomous and MITEs in the class II families. Interestingly, we found fragments of NLTRs corresponding to the RT domain, which preserves high identity among them, suggesting a possible remaining genomic role for these domains. Conclusions We show here that the TEs in the An. gambiae genome deteriorate in different ways according to the class to which they belong. This diversity certainly has implications not only at the host

  16. Synergy in efficacy of fungal entomopathogens and permethrin against West African insecticide-resistant Anopheles gambiae mosquitoes.

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    Marit Farenhorst

    Full Text Available BACKGROUND: Increasing incidences of insecticide resistance in malaria vectors are threatening the sustainable use of contemporary chemical vector control measures. Fungal entomopathogens provide a possible additional tool for the control of insecticide-resistant malaria mosquitoes. This study investigated the compatibility of the pyrethroid insecticide permethrin and two mosquito-pathogenic fungi, Beauveria bassiana and Metarhizium anisopliae, against a laboratory colony and field population of West African insecticide-resistant Anopheles gambiae s.s. mosquitoes. METHODOLOGY/FINDINGS: A range of fungus-insecticide combinations was used to test effects of timing and sequence of exposure. Both the laboratory-reared and field-collected mosquitoes were highly resistant to permethrin but susceptible to B. bassiana and M. anisopliae infection, inducing 100% mortality within nine days. Combinations of insecticide and fungus showed synergistic effects on mosquito survival. Fungal infection increased permethrin-induced mortality rates in wild An. gambiae s.s. mosquitoes and reciprocally, exposure to permethrin increased subsequent fungal-induced mortality rates in both colonies. Simultaneous co-exposure induced the highest mortality; up to 70.3+/-2% for a combined Beauveria and permethrin exposure within a time range of one gonotrophic cycle (4 days. CONCLUSIONS/SIGNIFICANCE: Combining fungi and permethrin induced a higher impact on mosquito survival than the use of these control agents alone. The observed synergism in efficacy shows the potential for integrated fungus-insecticide control measures to dramatically reduce malaria transmission and enable control at more moderate levels of coverage even in areas where insecticide resistance has rendered pyrethroids essentially ineffective.

  17. Synergy between repellents and non-pyrethroid insecticides strongly extends the efficacy of treated nets against Anopheles gambiae

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    N'Guessan Raphaël

    2007-03-01

    Full Text Available Abstract Background To manage the kdr pyrethroid-resistance in Anopheline malaria vectors, new compounds or new strategies are urgently needed. Recently, mixing repellents (DEET and a non-pyrethroid insecticide (propoxur was shown to be as effective as deltamethrin, a standard pyrethroid, under laboratory conditions, because of a strong synergy between the two compounds. In the present study, the interactions between two repellents (DEET and KBR 3023 and a non-pyrethroid insecticide (pyrimiphos methyl or PM on netting were investigated. The residual efficacy and the inhibition of blood feeding conferred by these mixtures were assessed against Anopheles gambiae mosquitoes. Methods DEET and KBR 3023 were mixed with pyrimiphos methyl (PM, a organophosphate (OP insecticide. The performance of mono- and bi-impregnated nets against adult mosquitoes was assessed using a miniaturized, experimental hut system (laboratory tunnel tests that allows expression of behavioural responses to insecticide, particularly the mortality and blood feeding effects. Results Both mixtures (PM+DEET and PM+KBR3023 induced 95% mortality for more than two months compared with less than one week for each compound used alone, then reflecting a strong synergy between the repellents and PM. A similar trend was observed with the blood feeding rates, which were significantly lower for the mixtures than for each component alone. Conclusion Synergistic interactions between organophosphates and repellents may be of great interest for vector control as they may contribute to increase the residual life of impregnated materials and improve the control of pyrethroid-resistance mosquitoes. These results prompt the need to evaluate the efficacy of repellent/non-pyrethroid insecticide mixtures against field populations of An. gambiae showing high level of resistance to Ops and pyrethroids.

  18. Comparative genomic analysis of chitinase and chitinase-like genes in the African malaria mosquito (Anopheles gambiae.

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    Jianzhen Zhang

    Full Text Available Chitinase is an important enzyme responsible for chitin metabolism in a wide range of organisms including bacteria, yeasts and other fungi, nematodes and arthropods. However, current knowledge on chitinolytic enzymes, especially their structures, functions and regulation is very limited. In this study we have identified 20 chitinase and chitinase-like genes in the African malaria mosquito, Anopheles gambiae, through genome-wide searching and transcript profiling. We assigned these genes into eight different chitinase groupings (groups I-VIII. Domain analysis of their predicted proteins showed that all contained at least one catalytic domain. However, only seven (AgCht4, AgCht5-1, AgCht6, AgCht7, AgCht8, AgCht10 and AgCht23 displayed one or more chitin-binding domains. Analyses of stage- and tissue-specific gene expression revealed that most of these genes were expressed in larval stages. However, AgCht8 was mainly expressed in the pupal and adult stages. AgCht2 and AgCht12 were specifically expressed in the foregut, whereas AgCht13 was only expressed in the midgut. The high diversity and complexity of An. gambiae chitinase and chitinase-like genes suggest their diverse functions during different developmental stages and in different tissues of the insect. A comparative genomic analysis of these genes along with those present in Drosophila melanogaster, Tribolium castaneum and several other insect species led to a uniform classification and nomenclature of these genes. Our investigation also provided important information for conducting future studies on the functions of chitinase and chitinase-like genes in this important malaria vector and other species of arthropods.

  19. A simplified high-throughput method for pyrethroid knock-down resistance (kdr detection in Anopheles gambiae

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    Walker Edward D

    2005-03-01

    Full Text Available Abstract Background A single base pair mutation in the sodium channel confers knock-down resistance to pyrethroids in many insect species. Its occurrence in Anopheles mosquitoes may have important implications for malaria vector control especially considering the current trend for large scale pyrethroid-treated bednet programmes. Screening Anopheles gambiae populations for the kdr mutation has become one of the mainstays of programmes that monitor the development of insecticide resistance. The screening is commonly performed using a multiplex Polymerase Chain Reaction (PCR which, since it is reliant on a single nucleotide polymorphism, can be unreliable. Here we present a reliable and potentially high throughput method for screening An. gambiae for the kdr mutation. Methods A Hot Ligation Oligonucleotide Assay (HOLA was developed to detect both the East and West African kdr alleles in the homozygous and heterozygous states, and was optimized for use in low-tech developing world laboratories. Results from the HOLA were compared to results from the multiplex PCR for field and laboratory mosquito specimens to provide verification of the robustness and sensitivity of the technique. Results and Discussion The HOLA assay, developed for detection of the kdr mutation, gives a bright blue colouration for a positive result whilst negative reactions remain colourless. The results are apparent within a few minutes of adding the final substrate and can be scored by eye. Heterozygotes are scored when a sample gives a positive reaction to the susceptible probe and the kdr probe. The technique uses only basic laboratory equipment and skills and can be carried out by anyone familiar with the Enzyme-linked immunosorbent assay (ELISA technique. A comparison to the multiplex PCR method showed that the HOLA assay was more reliable, and scoring of the plates was less ambiguous. Conclusion The method is capable of detecting both the East and West African kdr alleles

  20. Hydric stress-dependent effects of Plasmodium falciparum infection on the survival of wild-caught Anopheles gambiae female mosquitoes

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    Aboagye-Antwi Fred

    2010-08-01

    Full Text Available Abstract Background Whether Plasmodium falciparum, the agent of human malaria responsible for over a million deaths per year, causes fitness costs in its mosquito vectors is a burning question that has not yet been adequately resolved. Understanding the evolutionary forces responsible for the maintenance of susceptibility and refractory alleles in natural mosquito populations is critical for understanding malaria transmission dynamics. Methods In natural mosquito populations, Plasmodium fitness costs may only be expressed in combination with other environmental stress factors hence this hypothesis was tested experimentally. Wild-caught blood-fed Anopheles gambiae s.s. females of the M and S molecular form from an area endemic for malaria in Mali, West Africa, were brought to the laboratory and submitted to a 7-day period of mild hydric stress or kept with water ad-libitum. At the end of this experiment all females were submitted to intense desiccation until death. The survival of all females throughout both stress episodes, as well as their body size and infection status was recorded. The importance of stress, body size and molecular form on infection prevalence and female survival was investigated using Logistic Regression and Proportional-Hazard analysis. Results Females subjected to mild stress exhibited patterns of survival and prevalence of infection compatible with increased parasite-induced mortality compared to non-stressed females. Fitness costs seemed to be linked to ookinetes and early oocyst development but not the presence of sporozoites. In addition, when females were subjected to intense desiccation stress, those carrying oocysts exhibited drastically reduced survival but those carrying sporozoites were unaffected. No significant differences in prevalence of infection and infection-induced mortality were found between the M and S molecular forms of Anopheles gambiae. Conclusions Because these results suggest that infected

  1. Risk mapping of Anopheles gambiae s.l. densities using remotely-sensed environmental and meteorological data in an urban area: Dakar, Senegal.

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    Vanessa Machault

    Full Text Available INTRODUCTION: High malaria transmission heterogeneity in an urban environment is basically due to the complex distribution of Anopheles larval habitats, sources of vectors. Understanding 1 the meteorological and ecological factors associated with differential larvae spatio-temporal distribution and 2 the vectors dynamic, both may lead to improving malaria control measures with remote sensing and high resolution data as key components. In this study a robust operational methodology for entomological malaria predictive risk maps in urban settings is developed. METHODS: The Tele-epidemiology approach, i.e., 1 intensive ground measurements (Anopheles larval habitats and Human Biting Rate, or HBR, 2 selection of the most appropriate satellite data (for mapping and extracting environmental and meteorological information, and 3 use of statistical models taking into account the spatio-temporal data variability has been applied in Dakar, Senegal. RESULTS: First step was to detect all water bodies in Dakar. Secondly, environmental and meteorological conditions in the vicinity of water bodies favoring the presence of Anopheles gambiae s.l. larvae were added. Then relationship between the predicted larval production and the field measured HBR was identified, in order to generate An. gambiae s.l. HBR high resolution maps (daily, 10-m pixel in space. DISCUSSION AND CONCLUSION: A robust operational methodology for dynamic entomological malaria predictive risk maps in an urban setting includes spatio-temporal variability of An. gambiae s.l. larval habitats and An. gambiae s.l. HBR. The resulting risk maps are first examples of high resolution products which can be included in an operational warning and targeting system for the implementation of vector control measures.

  2. The angiotensin-converting enzyme (ACE) gene family of Anopheles gambiae.

    OpenAIRE

    Isaac R Elwyn; Lee Alison J; Smith Judith A; Burnham Susan; Shirras Alan D

    2005-01-01

    Abstract Background Members of the M2 family of peptidases, related to mammalian angiotensin converting enzyme (ACE), play important roles in regulating a number of physiological processes. As more invertebrate genomes are sequenced, there is increasing evidence of a variety of M2 peptidase genes, even within a single species. The function of these ACE-like proteins is largely unknown. Sequencing of the A. gambiae genome has revealed a number of ACE-like genes but probable errors in the Ensem...

  3. Fine pathogen discrimination within the APL1 gene family protects Anopheles gambiae against human and rodent malaria species.

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    Christian Mitri

    2009-09-01

    Full Text Available Genetically controlled resistance of Anopheles gambiae mosquitoes to Plasmodium falciparum is a common trait in the natural population, and a cluster of natural resistance loci were mapped to the Plasmodium-Resistance Island (PRI of the A. gambiae genome. The APL1 family of leucine-rich repeat (LRR proteins was highlighted by candidate gene studies in the PRI, and is comprised of paralogs APL1A, APL1B and APL1C that share > or =50% amino acid identity. Here, we present a functional analysis of the joint response of APL1 family members during mosquito infection with human and rodent Plasmodium species. Only paralog APL1A protected A. gambiae against infection with the human malaria parasite P. falciparum from both the field population and in vitro culture. In contrast, only paralog APL1C protected against the rodent malaria parasites P. berghei and P. yoelii. We show that anti-P. falciparum protection is mediated by the Imd/Rel2 pathway, while protection against P. berghei infection was shown to require Toll/Rel1 signaling. Further, only the short Rel2-S isoform and not the long Rel2-F isoform of Rel2 confers protection against P. falciparum. Protection correlates with the transcriptional regulation of APL1A by Rel2-S but not Rel2-F, suggesting that the Rel2-S anti-parasite phenotype results at least in part from its transcriptional control over APL1A. These results indicate that distinct members of the APL1 gene family display a mutually exclusive protective effect against different classes of Plasmodium parasites. It appears that a gene-for-pathogen-class system orients the appropriate host defenses against distinct categories of similar pathogens. It is known that insect innate immune pathways can distinguish between grossly different microbes such as Gram-positive bacteria, Gram-negative bacteria, or fungi, but the function of the APL1 paralogs reveals that mosquito innate immunity possesses a more fine-grained capacity to distinguish between

  4. Allomonal effect of breath contributes to differential attractiveness of humans to the African malaria vector Anopheles gambiae

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    Takken Willem

    2004-01-01

    Full Text Available Abstract Background Removal of exhaled air from total body emanations or artificially standardising carbon dioxide (CO2 outputs has previously been shown to eliminate differential attractiveness of humans to certain blackfly (Simuliidae and mosquito (Culicidae species. Whether or not breath contributes to between-person differences in relative attractiveness to the highly anthropophilic malaria vector Anopheles gambiae sensu stricto remains unknown and was the focus of the present study. Methods The contribution to and possible interaction of breath (BR and body odours (BO in the attraction of An. gambiae s.s. to humans was investigated by conducting dual choice tests using a recently developed olfactometer. Either one or two human subjects were used as bait. The single person experiments compared the attractiveness of a person's BR versus that person's BO or a control (empty tent with no odour. His BO and total emanations (TE = BR+BO were also compared with a control. The two-person experiments compared the relative attractiveness of their TE, BO or BR, and the TE of each person against the BO of the other. Results Experiments with one human subject (P1 as bait found that his BO and TE collected more mosquitoes than the control (P = 0.005 and P 1 attracted more mosquitoes than that of another person designated P8 (P 8 attracted more mosquitoes than the BR of P1 (P = 0.001. The attractiveness of the BO of P1 versus the BO of P8 did not differ (P = 0.346. The BO from either individual was consistently more attractive than the TE from the other (P Conclusions We demonstrated for the first time that human breath, although known to contain semiochemicals that elicit behavioural and/or electrophysiological responses (CO2, ammonia, fatty acids in An. gambiae also contains one or more constituents with allomonal (~repellent properties, which inhibit attraction and may serve as an important contributor to between-person differences in the relative

  5. Living at the edge: biogeographic patterns of habitat segregation conform to speciation by niche expansion in Anopheles gambiae

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    Costantini Carlo

    2009-05-01

    Full Text Available Abstract Background Ongoing lineage splitting within the African malaria mosquito Anopheles gambiae is compatible with ecological speciation, the evolution of reproductive isolation by divergent natural selection acting on two populations exploiting alternative resources. Divergence between two molecular forms (M and S identified by fixed differences in rDNA, and characterized by marked, although incomplete, reproductive isolation is occurring in West and Central Africa. To elucidate the role that ecology and geography play in speciation, we carried out a countrywide analysis of An. gambiae M and S habitat requirements, and that of their chromosomal variants, across Burkina Faso. Results Maps of relative abundance by geostatistical interpolators produced a distinct pattern of distribution: the M-form dominated in the northernmost arid zones, the S-form in the more humid southern regions. Maps of habitat suitability, quantified by Ecological Niche Factor Analysis based on 15 eco-geographical variables revealed less contrast among forms. M was peculiar as it occurred proportionally more in habitat of marginal quality. Measures of ecological niche breadth and overlap confirmed the mismatch between the fundamental and realized patterns of habitat occupation: forms segregated more than expected from the extent of divergence of their environmental envelope – a signature of niche expansion. Classification of chromosomal arm 2R karyotypes by multilocus genetic clustering identified two clusters loosely corresponding to molecular forms, with 'mismatches' representing admixed individuals due to shared ancestral polymorphism and/or residual hybridization. In multivariate ordination space, these karyotypes plotted in habitat of more marginal quality compared to non-admixed, 'typical', karyotypes. The distribution of 'typical' karyotypes along the main eco-climatic gradient followed a consistent pattern within and between forms, indicating an adaptive role

  6. Effect of larval crowding on mating competitiveness of Anopheles gambiae mosquitoes

    NARCIS (Netherlands)

    Ng'habi, K.R.; John, B.; Nkwengulila, G.; Knols, B.G.J.; Killeen, G.F.; Ferguson, H.M.

    2005-01-01

    Background: The success of sterile or transgenic Anopheles for malaria control depends on their mating competitiveness within wild populations. Current evidence suggests that transgenic mosquitoes have reduced fitness. One means of compensating for this fitness deficit would be to identify environme

  7. Annotation and analysis of a large cuticular protein family with the R&R Consensus in Anopheles gambiae

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    He Ningjia

    2008-01-01

    Full Text Available Abstract Background The most abundant family of insect cuticular proteins, the CPR family, is recognized by the R&R Consensus, a domain of about 64 amino acids that binds to chitin and is present throughout arthropods. Several species have now been shown to have more than 100 CPR genes, inviting speculation as to the functional importance of this large number and diversity. Results We have identified 156 genes in Anopheles gambiae that code for putative cuticular proteins in this CPR family, over 1% of the total number of predicted genes in this species. Annotation was verified using several criteria including identification of TATA boxes, INRs, and DPEs plus support from proteomic and gene expression analyses. Two previously recognized CPR classes, RR-1 and RR-2, form separate, well-supported clades with the exception of a small set of genes with long branches whose relationships are poorly resolved. Several of these outliers have clear orthologs in other species. Although both clades are under purifying selection, the RR-1 variant of the R&R Consensus is evolving at twice the rate of the RR-2 variant and is structurally more labile. In contrast, the regions flanking the R&R Consensus have diversified in amino-acid composition to a much greater extent in RR-2 genes compared with RR-1 genes. Many genes are found in compact tandem arrays that may include similar or dissimilar genes but always include just one of the two classes. Tandem arrays of RR-2 genes frequently contain subsets of genes coding for highly similar proteins (sequence clusters. Properties of the proteins indicated that each cluster may serve a distinct function in the cuticle. Conclusion The complete annotation of this large gene family provides insight on the mechanisms of gene family evolution and clues about the need for so many CPR genes. These data also should assist annotation of other Anopheles genes.

  8. Random amplified polymorphic DNA (RAPD) markers readily distinguish cryptic mosquito species (Diptera: Culicidae: Anopheles).

    Science.gov (United States)

    Wilkerson, R C; Parsons, T J; Albright, D G; Klein, T A; Braun, M J

    1993-01-01

    The usefulness of random amplified polymorphic DNA (RAPD) was examined as a potential tool to differentiate cryptic mosquito species. It proved to be a quick, effective means of finding genetic markers to separate two laboratory populations of morphologically indistinguishable African malaria vectors, Anopheles gambiae and An. arabiensis. In an initial screening of fifty-seven RAPD primers, 377 bands were produced, 295 of which differed between the two species. Based on criteria of interpretability, simplicity and reproducibility, thirteen primers were chosen for further screening using DNA from thirty individuals of each species. Seven primers produced diagnostic bands, five of which are described here. Some problematic characteristics of RAPD banding patterns are discussed and approaches to overcome these are suggested. PMID:8269099

  9. Insecticidal activity of petroleum ether extract and essential oil of Chenopodium ambrosioides L.(Chenopodiaceae) against Anopheles gambiae(Diptera: Culicidae)%土荆芥的石油醚提取物及精油对冈比亚按蚊的杀虫活性

    Institute of Scientific and Technical Information of China (English)

    Abiodun A. DENLOYE; Oluwakemi K. AJELARA; Rasaq A. OLOWU; Adeolu O. ESHILOKUN; Winifred A. MAKANJUOLA

    2009-01-01

    . ambrosioides for the control An. gambiae.%48 h致死中浓度(LC50)的测试结果表明,石油醚提取物对1龄幼虫的毒性最强(14.89 mg/L),其次是对4龄幼虫(18.90 mg/L),对3龄幼虫的毒性最低(183.77 mg/L); 精油对4龄幼虫毒性最强(36.62 mg/L),其次是对1龄幼虫(90.75 mg/L).推算的土荆芥精油对冈比亚按蚊的LC50为1.01 μL/L.本研究揭示了土荆芥对冈比亚按蚊的防治潜力.

  10. A possible mechanism for the suppression of Plasmodium berghei development in the mosquito Anopheles gambiae by the microsporidian Vavraia culicis.

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    Irka Bargielowski

    Full Text Available BACKGROUND: Microsporidian parasites of mosquitoes offer a possible way of controlling malaria, as they impede the development of Plasmodium parasites within the mosquito. The mechanism involved in this interference process is unknown. METHODOLOGY: We evaluated the possibility that larval infection by a microsporidian primes the immune system of adult mosquitoes in a way that enables a more effective anti-Plasmodium response. To do so, we infected 2-day old larvae of the mosquito Anopheles gambiae with one of 4 isolates of the microsporidian Vavraia culicis and reared one group as an uninfected control. Within each treatment, we fed half the adult females on a mix of P. berghei ookinetes and blood and inoculated the other half with a negatively charged CM-25 Sephadex bead to evaluate the mosquitoes' melanisation response. CONCLUSIONS: The microsporidian-infected mosquitoes were less likely to harbour oocysts (58.5% vs. 81.8%, harboured fewer oocysts (8.9 oocysts vs. 20.7 oocysts if the malaria parasite did develop and melanised the Sephadex bead to a greater degree (73% vs. 35% than the controls. While the isolates differed in the number of oocysts and in the melanisation response, the stimulation of the immune response was not correlated with either measure of malaria development. Nevertheless, the consistent difference between microsporidian-infected and -uninfected mosquitoes--more effective melanisation and less successful infection by malaria--suggests that microsporidians impede the development of malaria by priming the mosquito's immune system.

  11. Site-specific integration and expression of an anti-malarial gene in transgenic Anopheles gambiae significantly reduces Plasmodium infections.

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    Janet M Meredith

    Full Text Available Diseases transmitted by mosquitoes have a devastating impact on global health and this is worsening due to difficulties with existing control measures and climate change. Genetically modified mosquitoes that are refractory to disease transmission are seen as having great potential in the delivery of novel control strategies. Historically the genetic modification of insects has relied upon transposable elements which have many limitations despite their successful use. To circumvent these limitations the Streptomyces phage phiC31 integrase system has been successfully adapted for site-specific transgene integration in insects. Here, we present the first site-specific transformation of Anopheles gambiae, the principal vector of human malaria. Mosquitoes were initially engineered to incorporate the phiC31 targeting site at a defined genomic location. A second phase of genetic modification then achieved site-specific integration of Vida3, a synthetic anti-malarial gene. Expression of Vida3, specifically in the midgut of bloodfed females, offered consistent and significant protection against Plasmodium yoelii nigeriensis, reducing average parasite intensity by 85%. Similar protection was observed against Plasmodium falciparum in some experiments, although protection was inconsistent. In the fight against malaria, it is imperative to establish a broad repertoire of both anti-malarial effector genes and tissue-specific promoters for their expression, enabling those offering maximum effect with minimum fitness cost to be identified. In the future, this technology will allow effective comparisons and informed choices to be made, potentially leading to complete transmission blockade.

  12. A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae.

    Science.gov (United States)

    Hammond, Andrew; Galizi, Roberto; Kyrou, Kyros; Simoni, Alekos; Siniscalchi, Carla; Katsanos, Dimitris; Gribble, Matthew; Baker, Dean; Marois, Eric; Russell, Steven; Burt, Austin; Windbichler, Nikolai; Crisanti, Andrea; Nolan, Tony

    2016-01-01

    Gene drive systems that enable super-Mendelian inheritance of a transgene have the potential to modify insect populations over a timeframe of a few years. We describe CRISPR-Cas9 endonuclease constructs that function as gene drive systems in Anopheles gambiae, the main vector for malaria. We identified three genes (AGAP005958, AGAP011377 and AGAP007280) that confer a recessive female-sterility phenotype upon disruption, and inserted into each locus CRISPR-Cas9 gene drive constructs designed to target and edit each gene. For each targeted locus we observed a strong gene drive at the molecular level, with transmission rates to progeny of 91.4 to 99.6%. Population modeling and cage experiments indicate that a CRISPR-Cas9 construct targeting one of these loci, AGAP007280, meets the minimum requirement for a gene drive targeting female reproduction in an insect population. These findings could expedite the development of gene drives to suppress mosquito populations to levels that do not support malaria transmission. PMID:26641531

  13. A CRISPR-Cas9 gene drive system targeting female reproduction in the malaria mosquito vector Anopheles gambiae.

    Science.gov (United States)

    Hammond, Andrew; Galizi, Roberto; Kyrou, Kyros; Simoni, Alekos; Siniscalchi, Carla; Katsanos, Dimitris; Gribble, Matthew; Baker, Dean; Marois, Eric; Russell, Steven; Burt, Austin; Windbichler, Nikolai; Crisanti, Andrea; Nolan, Tony

    2016-01-01

    Gene drive systems that enable super-Mendelian inheritance of a transgene have the potential to modify insect populations over a timeframe of a few years. We describe CRISPR-Cas9 endonuclease constructs that function as gene drive systems in Anopheles gambiae, the main vector for malaria. We identified three genes (AGAP005958, AGAP011377 and AGAP007280) that confer a recessive female-sterility phenotype upon disruption, and inserted into each locus CRISPR-Cas9 gene drive constructs designed to target and edit each gene. For each targeted locus we observed a strong gene drive at the molecular level, with transmission rates to progeny of 91.4 to 99.6%. Population modeling and cage experiments indicate that a CRISPR-Cas9 construct targeting one of these loci, AGAP007280, meets the minimum requirement for a gene drive targeting female reproduction in an insect population. These findings could expedite the development of gene drives to suppress mosquito populations to levels that do not support malaria transmission.

  14. The interaction between a sexually transferred steroid hormone and a female protein regulates oogenesis in the malaria mosquito Anopheles gambiae.

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    Francesco Baldini

    2013-10-01

    Full Text Available Molecular interactions between male and female factors during mating profoundly affect the reproductive behavior and physiology of female insects. In natural populations of the malaria mosquito Anopheles gambiae, blood-fed females direct nutritional resources towards oogenesis only when inseminated. Here we show that the mating-dependent pathway of egg development in these mosquitoes is regulated by the interaction between the steroid hormone 20-hydroxy-ecdysone (20E transferred by males during copulation and a female Mating-Induced Stimulator of Oogenesis (MISO protein. RNAi silencing of MISO abolishes the increase in oogenesis caused by mating in blood-fed females, causes a delay in oocyte development, and impairs the function of male-transferred 20E. Co-immunoprecipitation experiments show that MISO and 20E interact in the female reproductive tract. Moreover MISO expression after mating is induced by 20E via the Ecdysone Receptor, demonstrating a close cooperation between the two factors. Male-transferred 20E therefore acts as a mating signal that females translate into an increased investment in egg development via a MISO-dependent pathway. The identification of this male-female reproductive interaction offers novel opportunities for the control of mosquito populations that transmit malaria.

  15. The Interaction between a Sexually Transferred Steroid Hormone and a Female Protein Regulates Oogenesis in the Malaria Mosquito Anopheles gambiae

    Science.gov (United States)

    Baldini, Francesco; Gabrieli, Paolo; South, Adam; Valim, Clarissa; Mancini, Francesca; Catteruccia, Flaminia

    2013-01-01

    Molecular interactions between male and female factors during mating profoundly affect the reproductive behavior and physiology of female insects. In natural populations of the malaria mosquito Anopheles gambiae, blood-fed females direct nutritional resources towards oogenesis only when inseminated. Here we show that the mating-dependent pathway of egg development in these mosquitoes is regulated by the interaction between the steroid hormone 20-hydroxy-ecdysone (20E) transferred by males during copulation and a female Mating-Induced Stimulator of Oogenesis (MISO) protein. RNAi silencing of MISO abolishes the increase in oogenesis caused by mating in blood-fed females, causes a delay in oocyte development, and impairs the function of male-transferred 20E. Co-immunoprecipitation experiments show that MISO and 20E interact in the female reproductive tract. Moreover MISO expression after mating is induced by 20E via the Ecdysone Receptor, demonstrating a close cooperation between the two factors. Male-transferred 20E therefore acts as a mating signal that females translate into an increased investment in egg development via a MISO-dependent pathway. The identification of this male–female reproductive interaction offers novel opportunities for the control of mosquito populations that transmit malaria. PMID:24204210

  16. Anopheles gambiae Ag-STAT, a new insect member of the STAT family, is activated in response to bacterial infection.

    Science.gov (United States)

    Barillas-Mury, C; Han, Y S; Seeley, D; Kafatos, F C

    1999-02-15

    A new insect member of the STAT family of transcription factors (Ag-STAT) has been cloned from the human malaria vector Anopheles gambiae. The domain involved in DNA interaction and the SH2 domain are well conserved. Ag-STAT is most similar to Drosophila D-STAT and to vertebrate STATs 5 and 6, constituting a proposed ancient class A of the STAT family. The mRNA is expressed at all developmental stages, and the protein is present in hemocytes, pericardial cells, midgut, skeletal muscle and fat body cells. There is no evidence of transcriptional activation following bacterial challenge. However, bacterial challenge results in nuclear translocation of Ag-STAT protein in fat body cells and induction of DNA-binding activity that recognizes a STAT target site. In vitro treatment with pervanadate (vanadate and H2O2) translocates Ag-STAT to the nucleus in midgut epithelial cells. This is the first evidence of direct participation of the STAT pathway in immune responses in insects. PMID:10022838

  17. Anopheles gambiae collagen IV genes: cloning, phylogeny and midgut expression associated with blood feeding and Plasmodium infection.

    Science.gov (United States)

    Gare, D C; Piertney, S B; Billingsley, P F

    2003-07-01

    A prerequisite for understanding the role that mosquito midgut extracellular matrix molecules play in malaria parasite development is proper isolation and characterisation of the genes coding for components of the basal lamina. Here we have identified genes coding for alpha1 and alpha2 chains of collagen IV from the major malaria vector, Anopheles gambiae. Conserved sequences in the terminal NC1 domain were used to obtain partial gene sequences of this functional region, and full sequence was isolated from a pupal cDNA library. In a DNA-derived phylogeny, the alpha1 and alpha2 chains cluster with dipteran orthologs, and the alpha2 is ancestral. The expression of collagen alpha1(IV) peaked during the pupal stage of mosquito development, and was expressed continuously in the adult female following a blood meal with a further rise detected in older mosquitoes. Collagen alpha1(IV) is also upregulated when the early oocyst of Plasmodium yoelii was developing within the mosquito midgut and may contribute to a larger wound healing response. A model describing the expression of basal lamina proteins during oocyst development is presented, and we hypothesise that the development of new basal lamina between the oocyst and midgut epithelium is akin to a wound healing process. PMID:12814648

  18. Apolipophorin-III Mediates Antiplasmodial Epithelial Responses in Anopheles gambiae (G3) Mosquitoes

    OpenAIRE

    Gupta, Lalita; Noh, Ju Young; Jo, Yong Hun; Oh, Seung Han; Kumar, Sanjeev; Noh, Mi Young; Lee, Yong Seok; Cha, Sung-Jae; Seo, Sook Jae; Kim, Iksoo; Han, Yeon Soo; Barillas-Mury, Carolina

    2010-01-01

    Background Apolipophorin-III (ApoLp-III) is known to play an important role in lipid transport and innate immunity in lepidopteran insects. However, there is no evidence of involvement of ApoLp-IIIs in the immune responses of dipteran insects such as Drosophila and mosquitoes. Methodology/Principal Findings We report the molecular and functional characterization of An. gambiae apolipophorin-III (AgApoLp-III). Mosquito ApoLp-IIIs have diverged extensively from those of lepidopteran insects; ho...

  19. Chemical composition and insecticidal activity of plant essential oils from Benin against Anopheles gambiae (Giles)

    OpenAIRE

    Bossou, Annick; Mangelinckx, Sven; Yedomonhan, Hounnankpon; Boko, Pelagie M; Akogbeto, Martin C; De Kimpe, Norbert; Avlessi, Felicien; Sohounhloue, Dominique CK

    2013-01-01

    Background Insecticide resistance in sub-Saharan Africa and especially in Benin is a major public health issue hindering the control of the malaria vectors. Each Anopheles species has developed a resistance to one or several classes of the insecticides currently in use in the field. Therefore, it is urgent to find alternative compounds to conquer the vector. In this study, the efficacies of essential oils of nine plant species, which are traditionally used to avoid mosquito bites in Benin, we...

  20. The neotype of anopheles albitarsis (Diptera: culicidae O neótipo de Anopheles albitarsis (Diptera: Culicidae

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    Maria Goreti Rosa-Freitas

    1989-09-01

    Full Text Available Anopheles albitarsis neotype is described from specimens collected in Baradero, Argentina, in Shannon's trap, in horse and pig stables and on the progeny of engorded females. The description includes illustrations of adult female, male and female genitalias, scanning electron miscroscopy of the eggs and complete chaetotaxy of pupa and larva. The importance for electing a neotype is based on the realization that An. albitarsis is a complex of cryptic species. It is an attempt to provide typt-locality specimens with which other memebers of the group can be compared.O neótipo de Anopheles albitarsis é descrito a partir de espécimens coletados em armadilha tipo Shannon, em estábulos de cavalos e porcos e progênies de fêmeas ingurgitadas em Baradero, Argentina, localidade-tipo da espécie. A descrição inclui ilustrações da fêmea adulda, genitálias masculina e feminina, ovos em microscopia eletrônica de varredura e da quetotaxia completa das larvas de 4º estádio e pupas. A eleição de um neótipo para albitarsis baseia-se em dados recentes que apontam a espécie como um complexo de espécies crípticas, o que evidencia a importância de uma descrição detalhada de espécimens da localidade-tipo com o qual outros membros do grupo possam ser comparados.

  1. Resistance Mechanisms of Anopheles stephensi (Diptera: Culicidae to Temephos

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    Aboozar Soltani

    2015-10-01

    Full Text Available Background: Anopheles stephensi is a sub-tropical species and has been considered as one of the most important vector of human malaria throughout the Middle East and South Asian region including the malarious areas of southern Iran. Current reports confirmed An. stephensi resistance to temephos in Oman and India. However, there is no comprehensive research on mechanisms of temephos resistance in An. stephensi in the literature. This study was designed in order to clarify the enzymatic and molecular mechanisms of temephos resistance in this species.Methods: Profile activities of α- and ß-esterases, mixed function oxidase (MFO, glutathione-S-transferase (GST, insensitive acetylcholinesterase, and para-nitrophenyl acetate (PNPA-esterase enzymes were tested for An. stephensi strain with resistance ratio of 15.82 to temephos in comparison with susceptible strain.Results: Results showed that the mean activity of α-EST, GST and AChE enzymes were classified as altered indicating metabolic mechanisms have considerable role in resistance of An. stephensi to temephos. Molecular study using PCR-RFLP method to trace the G119S mutation in ACE-1 gene showed lack of the mutation responsible for organophosphate insecticide resistance in the temephos-selected strain of An. stephensi.Conclusion: This study showed that the altered enzymes but not targets site insensitivity of ACE-1 are responsible for temephos resistance in An. stephensi in south of Iran.

  2. Description of the egg of Anopheles (Anopheles intermedius (Peryassu, 1908 (Diptera: Culicidae by scanning electron microscopy

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    Oswaldo Paulo Forattini

    1997-01-01

    Full Text Available The egg of Anopheles (Anopheles intermedius (Peryassu, 1908 is described and illustrated with scanning electron micrographs. Literature data on An. (Ano. maculipes (Theobald, 1903 is providedOs ovos de An. intermedius foram descritos e ilustrados por Costa Lima (1929. Este autor, baseando-se nos desenhos de Peryassu (1908 para An. maculipes, chamou atenção para o fato do ovo desta espécie ser semelhante ao de An. maculipes. Posteriormente, Causey e col. (1944, estudando os ovos de An. intermedius e An. maculipes ao estereomicroscópio, diferenciou-os por caracteres da franja. Em An. intermedius a franja é oblíqua ao eixo longitudinal do ovo, mas perpendicular em An. maculipes. Causey e col. (1944 ilustraram as variações morfológicas que encontraram na franja do ovo de An. intermedius. Os autores observaram que a franja apresentava-se descotínua em alguns espécimens de An. intermedius, tornando-se em pequenos círculos nessa região. Embora semelhante ao de An. maculipes, os ovos de An. intermedius podem ser facilmente reconhecidos por características da franja, flutuadores e tubérculos lobados, como pode ser observado na descrição. Contudo, será necessário estudar a morfologia dos ovos de outras espécies do subgênero para que se possa estabelecer as diferenças e fazer comparações precisas entre as espécies

  3. Next-generation site-directed transgenesis in the malaria vector mosquito Anopheles gambiae: self-docking strains expressing germline-specific phiC31 integrase.

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    Janet M Meredith

    Full Text Available Diseases transmitted by mosquitoes have a devastating impact on global health and the situation is complicated due to difficulties with both existing control measures and the impact of climate change. Genetically modified mosquitoes that are refractory to disease transmission are seen as having great potential in the delivery of novel control strategies. The Streptomyces phage phiC31 integrase system has been successfully adapted for site-directed transgene integration in a range of insects, thus overcoming many limitations due to size constraints and random integration associated with transposon-mediated transformation. Using this technology, we previously published the first site-directed transformation of Anopheles gambiae, the principal vector of human malaria. Mosquitoes were initially engineered to incorporate the phiC31 docking site at a defined genomic location. A second phase of genetic modification then achieved site-directed integration of an anti-malarial effector gene. In the current publication we report improved efficiency and utility of the phiC31 integrase system following the generation of Anopheles gambiae self-docking strains. Four independent strains, with docking sites at known locations on three different chromosome arms, were engineered to express integrase under control of the regulatory regions of the nanos gene from Anopheles gambiae. The resulting protein accumulates in the posterior oocyte to provide integrase activity at the site of germline development. Two self-docking strains, exhibiting significantly different levels of integrase expression, were assessed for site-directed transgene integration and found to demonstrate greatly improved survival and efficiency of transformation. In the fight against malaria, it is imperative to establish a broad repertoire of both anti-malarial effector genes and tissue-specific promoters to regulate their expression, enabling those offering maximum effect with minimum fitness

  4. Optimization of sugar and blood feeding regimen in Anopheles gambiae mass production system

    International Nuclear Information System (INIS)

    The sterile insect technique (SIT) is being developed for the control of malaria transmitting mosquitoes. Critical to the success of applying the SIT is the establishment of standardized mass production systems for the target species. As part of efforts to develop standardised mass production systems for malaria vectors, this project sought to optimize adult blood and sugar feeding in a mass production system. Different sugar types (glucose, sucrose and honey) were evaluated at 6% and 10% concentrations in water to determine the best sugar diet and concentration for feeding adult An. gambiae. Different blood feeding methods, restrained Guinea pig, anaesthetised Guinea pig and human arm feeding were evaluated. Adult survival, female insemination and egg production were used as criteria to determine optimum sugar and blood feeding. The effect of anaesthetics on blood feeding response and egg production of female An. gambiae was determined by comparing feeding response and egg production of females fed with anaesthetised Guinea pigs as against physically restrained Guinea pigs (Control). The specific effect of different anaesthetic agents on blood feeding response and egg production of female mosquitoes were was also determined by comparing the feeding response and egg production of females fed with either Ketamine/Xylazine anaesthetised Guinea pigs or Ketamine/Diazepam anaesthetised Guinea pigs. Effects due to sugar types and concentrations on percentage survival of male and female mosquitoes were observed to be significant at (p 0.05). However, human ann feeding (HAP) method and Ketamine/Xylazine (KX) anaesthetics fed for 25 minutes recorded higher percentage feeding (76.0% and 68.0% respectively) and egg production of 19.0% and 20.8% respectively. Anaesthetised Guinea pig feeding (AGF) of adults for 15 minutes followed closely with 60.0% and 15.1% blood feeding and egg production respectively whilst restrained Guinea pig feeding (RGF) method and Ketamine

  5. Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s. from Southern Benin and Nigeria

    OpenAIRE

    Ranson Hilary; Akogbeto Martin C; Coulibaly Ousmane N; Bakare Adekunle A; Djouaka Rousseau F; Hemingway Janet; Strode Clare

    2008-01-01

    Abstract Background Insecticide resistance in Anopheles mosquitoes is threatening the success of malaria control programmes. This is particularly true in Benin where pyrethroid resistance has been linked to the failure of insecticide treated bed nets. The role of mutations in the insecticide target sites in conferring resistance has been clearly established. In this study, the contribution of other potential resistance mechanisms was investigated in Anopheles gambiae s.s. from a number of loc...

  6. Molecular characterisation and chromosomal mapping of transcripts having tissue-specific expression in the malaria mosquito Anopheles gambiae: possible involvement in visual or olfactory processes.

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    Ricci, Irene; Santolamazza, Federica; Costantini, Carlo; Favia, Guido

    2002-01-01

    We have compared the transcriptional activity of heads, antennae + palps, and carcasses in the mosquito Anopheles gambiae by means of differential display PCR (DD-PCR). Three transcripts specifically or preferentially expressed in the heads and in the antennae + palps have been selected. All are very similar to genes related to visual and olfactory mechanisms of several different organisms. They have been named Ag arrestin, Ag rLDL, and Ag dynamin. The potential of the DD-PCR technique in identifying genes involved in mosquito behaviour and the usefulness of the molecular characterisation of these transcripts are discussed. PMID:11822731

  7. Pyrethroid resistance in Anopheles gambiae leads to increased susceptibility to the entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana

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    Knols Bart GJ

    2010-06-01

    Full Text Available Abstract Background Entomopathogenic fungi are being investigated as a new mosquito control tool because insecticide resistance is preventing successful mosquito control in many countries, and new methods are required that can target insecticide-resistant malaria vectors. Although laboratory studies have previously examined the effects of entomopathogenic fungi against adult mosquitoes, most application methods used cannot be readily deployed in the field. Because the fungi are biological organisms it is important to test potential field application methods that will not adversely affect them. The two objectives of this study were to investigate any differences in fungal susceptibility between an insecticide-resistant and insecticide-susceptible strain of Anopheles gambiae sensu stricto, and to test a potential field application method with respect to the viability and virulence of two fungal species Methods Pieces of white polyester netting were dipped in Metarhizium anisopliae ICIPE-30 or Beauveria bassiana IMI391510 mineral oil suspensions. These were kept at 27 ± 1°C, 80 ± 10% RH and the viability of the fungal conidia was recorded at different time points. Tube bioassays were used to infect insecticide-resistant (VKPER and insecticide-susceptible (SKK strains of An. gambiae s.s., and survival analysis was used to determine effects of mosquito strain, fungus species or time since fungal treatment of the net. Results The resistant VKPER strain was significantly more susceptible to fungal infection than the insecticide-susceptible SKK strain. Furthermore, B. bassiana was significantly more virulent than M. anisopliae for both mosquito strains, although this may be linked to the different viabilities of these fungal species. The viability of both fungal species decreased significantly one day after application onto polyester netting when compared to the viability of conidia remaining in suspension. Conclusions The insecticide

  8. The angiotensin-converting enzyme (ACE gene family of Anopheles gambiae

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    Isaac R Elwyn

    2005-12-01

    Full Text Available Abstract Background Members of the M2 family of peptidases, related to mammalian angiotensin converting enzyme (ACE, play important roles in regulating a number of physiological processes. As more invertebrate genomes are sequenced, there is increasing evidence of a variety of M2 peptidase genes, even within a single species. The function of these ACE-like proteins is largely unknown. Sequencing of the A. gambiae genome has revealed a number of ACE-like genes but probable errors in the Ensembl annotation have left the number of ACE-like genes, and their structure, unclear. Results TBLASTN and sequence analysis of cDNAs revealed that the A. gambiae genome contains nine genes (AnoACE genes which code for proteins with similarity to mammalian ACE. Eight of these genes code for putative single domain enzymes similar to other insect ACEs described so far. AnoACE9, however, has several features in common with mammalian somatic ACE such as a two domain structure and a hydrophobic C terminus. Four of the AnoACE genes (2, 3, 7 and 9 were shown to be expressed at a variety of developmental stages. Expression of AnoACE3, AnoACE7 and AnoACE9 is induced by a blood meal, with AnoACE7 showing the largest (approximately 10-fold induction. Conclusion Genes coding for two-domain ACEs have arisen several times during the course of evolution suggesting a common selective advantage to having an ACE with two active-sites in tandem in a single protein. AnoACE7 belongs to a sub-group of insect ACEs which are likely to be membrane-bound and which have an unusual, conserved gene structure.

  9. An epithelial serine protease, AgESP, is required for Plasmodium invasion in the mosquito Anopheles gambiae.

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    Janneth Rodrigues

    Full Text Available BACKGROUND: Plasmodium parasites need to cross the midgut and salivary gland epithelia to complete their life cycle in the mosquito. However, our understanding of the molecular mechanism and the mosquito genes that participate in this process is still very limited. METHODOLOGY/PRINCIPAL FINDINGS: We identified an Anopheles gambiae epithelial serine protease (AgESP that is constitutively expressed in the submicrovillar region of mosquito midgut epithelial cells and in the basal side of the salivary glands that is critical for Plasmodium parasites to cross these two epithelial barriers. AgESP silencing greatly reduces Plasmodium berghei and Plasmodium falciparum midgut invasion and prevents the transcriptional activation of gelsolin, a key regulator of actin remodeling and a reported Plasmodium agonist. AgESP expression is highly induced in midgut cells invaded by Plasmodium, suggesting that this protease also participates in the apoptotic response to invasion. In salivary gland epithelial cells, AgESP is localized on the basal side--the surface with which sporozoites interact. AgESP expression in the salivary gland is also induced in response to P. berghei and P. falciparum sporozoite invasion, and AgESP silencing significantly reduces the number of sporozoites that invade this organ. CONCLUSION: Our findings indicate that AgESP is required for Plasmodium parasites to effectively traverse the midgut and salivary gland epithelial barriers. Plasmodium parasites need to modify the actin cytoskeleton of mosquito epithelial cells to successfully complete their life cycle in the mosquito and AgESP appears to be a major player in the regulation of this process.

  10. An Epithelial Serine Protease, AgESP, Is Required for Plasmodium Invasion in the Mosquito Anopheles gambiae

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    Rodrigues, Janneth; Oliveira, Giselle A.; Kotsyfakis, Michalis; Dixit, Rajnikant; Molina-Cruz, Alvaro; Jochim, Ryan; Barillas-Mury, Carolina

    2012-01-01

    Background Plasmodium parasites need to cross the midgut and salivary gland epithelia to complete their life cycle in the mosquito. However, our understanding of the molecular mechanism and the mosquito genes that participate in this process is still very limited. Methodology/Principal Findings We identified an Anopheles gambiae epithelial serine protease (AgESP) that is constitutively expressed in the submicrovillar region of mosquito midgut epithelial cells and in the basal side of the salivary glands that is critical for Plasmodium parasites to cross these two epithelial barriers. AgESP silencing greatly reduces Plasmodium berghei and Plasmodium falciparum midgut invasion and prevents the transcriptional activation of gelsolin, a key regulator of actin remodeling and a reported Plasmodium agonist. AgESP expression is highly induced in midgut cells invaded by Plasmodium, suggesting that this protease also participates in the apoptotic response to invasion. In salivary gland epithelial cells, AgESP is localized on the basal side–the surface with which sporozoites interact. AgESP expression in the salivary gland is also induced in response to P. berghei and P. falciparum sporozoite invasion, and AgESP silencing significantly reduces the number of sporozoites that invade this organ. Conclusion Our findings indicate that AgESP is required for Plasmodium parasites to effectively traverse the midgut and salivary gland epithelial barriers. Plasmodium parasites need to modify the actin cytoskeleton of mosquito epithelial cells to successfully complete their life cycle in the mosquito and AgESP appears to be a major player in the regulation of this process. PMID:22509400

  11. The mode of action of spatial repellents and their impact on vectorial capacity of Anopheles gambiae sensu stricto.

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    Sheila B Ogoma

    Full Text Available Malaria vector control relies on toxicity of insecticides used in long lasting insecticide treated nets and indoor residual spraying. This is despite evidence that sub-lethal insecticides reduce human-vector contact and malaria transmission. The impact of sub-lethal insecticides on host seeking and blood feeding of mosquitoes was measured. Taxis boxes distinguished between repellency and attraction inhibition of mosquitoes by measuring response of mosquitoes towards or away from Transfluthrin coils and humans. Protective effective distance of coils and long-term effects on blood feeding were measured in the semi-field tunnel and in a Peet Grady chamber. Laboratory reared pyrethroid susceptible Anopheles gambiae sensu stricto mosquitoes were used. In the taxis boxes, a higher proportion of mosquitoes (67%-82% were activated and flew towards the human in the presence of Transfluthrin coils. Coils did not hinder attraction of mosquitoes to the human. In the semi-field Tunnel, coils placed 0.3 m from the human reduced feeding by 86% (95% CI [0.66; 0.95] when used as a "bubble" compared to 65% (95% CI [0.51; 0.76] when used as a "point source". Mosquitoes exposed to coils inside a Peet Grady chamber were delayed from feeding normally for 12 hours but there was no effect on free flying and caged mosquitoes exposed in the semi-field tunnel. These findings indicate that airborne pyrethroids minimize human-vector contact through reduced and delayed blood feeding. This information is useful for the development of target product profiles of spatial repellent products that can be used to complement mainstream malaria vector control tools.

  12. Effect of three larval diets on larval development and male sexual performance of Anopheles gambiae s.s.

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    Yahouédo, Gildas A; Djogbénou, Luc; Saïzonou, Jacques; Assogba, Bénoît S; Makoutodé, Michel; Gilles, Jeremie R L; Maïga, Hamidou; Mouline, Karine; Soukou, Bhonna K; Simard, Frédéric

    2014-04-01

    Population replacement/elimination strategies based on mass-release of sterile or otherwise genetically modified (male) mosquitoes are being considered in order to expand the malaria vector control arsenal on the way to eradication. A challenge in this context, is to produce male mosquitoes that will be able to compete and mate with wild females more efficiently than their wild counterparts, i.e. high fitness males. This study explored the effect of three larval food diets developed by the International Atomic Energy Agency on the overall fitness and mating performance of male Anopheles gambiae s.s. mosquitoes (Kisumu strain). Larval development (pupation and emergence rate, development time) was monitored, and adult wing length and energy reserves at emergence (i.e. lipids, sugars, glycogen and proteins) were measured. Male sexual performance was assessed through an insemination test whereby one male and 10 virgin females were maintained together in the same cage in order to record the number of inseminated females per 24h. Our results show that males reared on Diets 2 and 3 performed best during larval development. Males provided with treatment 2.2 had a shorter development time and performed best in insemination tests. However, these males had the lowest overall lifespan, suggesting a trade-off between longevity and sexual performances which needs to be taken into consideration when planning release. The results from this work were discussed in the context of sterile insect techniques or genetic control methods which is today one of the strategy in the overall mosquito control and elimination efforts. PMID:24291460

  13. Sensitivity of Anopheles gambiae population dynamics to meteo-hydrological variability: a mechanistic approach

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    Gilioli Gianni

    2011-10-01

    Full Text Available Abstract Background Mechanistic models play an important role in many biological disciplines, and they can effectively contribute to evaluate the spatial-temporal evolution of mosquito populations, in the light of the increasing knowledge of the crucial driving role on vector dynamics played by meteo-climatic features as well as other physical-biological characteristics of the landscape. Methods In malaria eco-epidemiology landscape components (atmosphere, water bodies, land use interact with the epidemiological system (interacting populations of vector, human, and parasite. In the background of the eco-epidemiological approach, a mosquito population model is here proposed to evaluate the sensitivity of An. gambiae s.s. population to some peculiar thermal-pluviometric scenarios. The scenarios are obtained perturbing meteorological time series data referred to four Kenyan sites (Nairobi, Nyabondo, Kibwesi, and Malindi representing four different eco-epidemiological settings. Results Simulations highlight a strong dependence of mosquito population abundance on temperature variation with well-defined site-specific patterns. The upper extreme of thermal perturbation interval (+ 3°C gives rise to an increase in adult population abundance at Nairobi (+111% and Nyabondo (+61%, and a decrease at Kibwezi (-2% and Malindi (-36%. At the lower extreme perturbation (-3°C is observed a reduction in both immature and adult mosquito population in three sites (Nairobi -74%, Nyabondo -66%, Kibwezi -39%, and an increase in Malindi (+11%. A coherent non-linear pattern of population variation emerges. The maximum rate of variation is +30% population abundance for +1°C of temperature change, but also almost null and negative values are obtained. Mosquitoes are less sensitive to rainfall and both adults and immature populations display a positive quasi-linear response pattern to rainfall variation. Conclusions The non-linear temperature-dependent response is in

  14. Comparative susceptibility to permethrin of two Anopheles gambiae s.l. populations from Southern Benin, regarding mosquito sex, physiological status, and mosquito age

    Institute of Scientific and Technical Information of China (English)

    Nazaire Azoun; Rock Akpon; Roseric Azondekon; Alex Asidi; Martin Akogbto

    2014-01-01

    Objective: To investigate what kind of mosquito sample is necessary for the determination of insecticide susceptibility in malaria vectors. Methods:Larvae and pupae of Anopheles gambiae s.l. (An. gambiae) mosquitoes were collected from the breeding sites in Littoral and Oueme departments. The Centers for Disease Control and Prevention (CDC) susceptibility tests were conducted on unfed male and female mosquitoes aged 2-5 days old. CDC susceptibility tests were also conducted on unfed, blood fed and gravid female mosquitoes aged 2-5 days old. These susceptibility tests were also conducted on unfed and blood fed female mosquitoes aged 2-5 days old and 20 days old. CDC biochemical assay using synergist was also carried out to detect any increase in the activity of enzyme typically involved in insecticide metabolism. Results:Female An. gambiae Ladji and Sekandji populations were more susceptible than the males when they were unfed and aged 2-5 days old. The mortality rates of blood fed female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when females were unfed. In addition, the mortality rates of gravid female An. gambiae Ladji and Sekandji populations aged 2-5 days old were lower than those obtained when they were unfed. The mortality rate obtained when female An. gambiae Sekandji populations were unfed and aged 20 days old was higher than the one obtained when these populations were unfed and aged 2-5 days old. The results obtained after effects of synergist penicillin in beeswax on F1 progeny of An. gambiae Ladji populations resistant to permethrin showed that mono-oxygenases were involved in permethrin resistant F1 progeny from Ladji. Conclusions: The resistance is a hereditary and dynamic phenomenon which can be due to metabolic mechanisms like overproduction of detoxifying enzymes activity. Many factors influence vector susceptibility to insecticide. Among these factors, there are mosquito sex, mosquito age, its

  15. The vasa regulatory region mediates germline expression and maternal transmission of proteins in the malaria mosquito Anopheles gambiae: a versatile tool for genetic control strategies

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    Burt Austin

    2009-07-01

    Full Text Available Abstract Background Germline specific promoters are an essential component of potential vector control strategies which function by genetic drive, however suitable promoters are not currently available for the main human malaria vector Anopheles gambiae. Results We have identified the Anopheles gambiae vasa-like gene and found its expression to be specifically localized to both the male and female gonads in adult mosquitoes. We have functionally characterised using transgenic reporter lines the regulatory regions required for driving transgene expression in a pattern mirroring that of the endogenous vasa locus. Two reporter constructs indicate the existence of distinct vasa regulatory elements within the 5' untranslated regions responsible not only for the spatial and temporal but also for the sex specific germline expression. vasa driven eGFP expression in the ovary of heterozygous mosquitoes resulted in the progressive accumulation of maternal protein and transcript in developing oocytes that were then detectable in all embryos and neonatal larvae. Conclusion We have characterized the vasa regulatory regions that are not only suited to drive transgenes in the early germline of both sexes but could also be utilized to manipulate the zygotic genome of developing embryos via maternal deposition of active molecules. We have used computational models to show that a homing endonuclease-based gene drive system can function in the presence of maternal deposition and describe a novel non-invasive control strategy based on early vasa driven homing endonuclease expression.

  16. Insecticide resistance in Anopheles gambiae: data from the first year of a multi-country study highlight the extent of the problem

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    Sagnon N'Falé

    2009-12-01

    Full Text Available Abstract Background Insecticide resistance in malaria vectors is a growing concern in many countries which requires immediate attention because of the limited chemical arsenal available for vector control. The current extent and distribution of this resistance in many parts of the continent is unknown and yet such information is essential for the planning of effective malaria control interventions. Methods In 2008, a network was established, with financial support from WHO/TDR, to investigate the extent of insecticide resistance in malaria vectors in five African countries. Here, the results of bioassays on Anopheles gambiae sensu lato from two rounds of monitoring from 12 sentinel sites in three of the partner countries are reported. Results Resistance is very heterogeneous even over relatively small distances. Furthermore, in some sites, large differences in mortality rates were observed during the course of the malaria transmission season. Using WHO diagnostic doses, all populations from Burkina Faso and Chad and two of the four populations from Sudan were classified as resistant to permethrin and/or deltamethrin. Very high frequencies of DDT resistance were found in urban areas in Burkina Faso and Sudan and in a cotton-growing district in Chad. In areas where both An. gambiae s.s. and Anopheles arabiensis were present, resistance was found in both species, although generally at a higher frequency in An gambiae s.s. Anopheles gambiae s.l. remains largely susceptible to the organophosphate fenitrothion and the carbamate bendiocarb in the majority of the sentinel sites with the exception of two sites in Burkina Faso. In the cotton-growing region of Soumousso in Burkina Faso, the vector population is resistant to all four classes of insecticide available for malaria control. Conclusions Possible factors influencing the frequency of resistant individuals observed in the sentinel sites are discussed. The results of this study highlight the

  17. Evidence of increasing L1014F kdr mutation frequency in Anopheles gambiae s.l. pyrethroid resistant following a nationwide distribution of LLINs by the Beninese National Malaria Control Programme

    Institute of Scientific and Technical Information of China (English)

    Nazaire Azoun; Rock Akpon; Martin Akogbto

    2014-01-01

    Objective:To determine the susceptibility status to pyrethroid in Anopheles gambiae s.l. (An. gambiae), the distribution of kdr“Leu-Phe”mutation in malaria vectors in Benin and to compare the current frequency of kdr“Leu-Phe”mutation to the previous frequency after long-lasting insecticide treated nets implementation. Methods: Larvae and pupae of An. gambiae s.l. mosquitoes were collected from the breeding sites in Littoral, Zou, Borgou and Alibori provinces. CDC susceptibility tests were conducted on unfed females mosquitoes aged 2-5 d old. An. gambiae mosquitoes were identified to species using PCR techniques. Molecular assays were also carried out to identify kdr mutations in individual mosquitoes. Results: The results showed that An. gambiae Malanville and Suru-lere populations were resistant to deltamethrin. Regarding An. gambiae Parakou and Bohicon populations, they were resistant to permethrin. PCR revealed 100%of mosquitoes tested were An. gambiae s.s. The L1014F kdr mutation was found in An. gambiae s.s. Malanville and Parakou at various allelic frequencies. The increase of kdr allelic frequency was positively correlated with CDC bioassays data. Conclusions: Pyrethroid resistance is widespread in malaria vector in Benin and kdr mutation is the main resistance mechanism involved. More attention may be paid for the future success of malaria control programmes based on LLINs with pyrethroids in the country.

  18. The evolution of TEP1, an exceptionally polymorphic immunity gene in Anopheles gambiae

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    Yan Guiyun

    2008-10-01

    Full Text Available Abstract Background Host-parasite coevolution can result in balancing selection, which maintains genetic variation in the susceptibility of hosts to parasites. It has been suggested that variation in a thioester-containing protein called TEP1 (AGAP010815 may alter the ability of Anopheles mosquitoes to transmit Plasmodium parasites, and high divergence between alleles of this gene suggests the possible action of long-term balancing selection. We studied whether TEP1 is a case of an ancient balanced polymorphism in an animal immune system. Results We found evidence that the high divergence between TEP1 alleles is the product of genetic exchange between TEP1 and other TEP loci, i.e. gene conversion. Additionally, some TEP1 alleles showed unexpectedly low variability. Conclusion The TEP1 gene appears to be a chimera produced from at least two other TEP loci, and the divergence between TEP1 alleles is probably not caused by long-term balancing selection, but is instead due to two independent gene conversion events from one of these other genes. Nevertheless, TEP1 still shows evidence of natural selection, in particular there appears to have been recent changes in the frequency of alleles that has diminished polymorphism within each allelic class. Although the selective force driving this dynamic was not identified, given that susceptibility to Plasmodium parasites is known to be associated with allelic variation in TEP1, these changes in allele frequencies could alter the vectoring capacity of populations.

  19. 3-Oxoisoxazole-2(3H)-carboxamides and isoxazol-3-yl carbamates: Resistance-breaking acetylcholinesterase inhibitors targeting the malaria mosquito, Anopheles gambiae

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    Verma, Astha; Wong, Dawn M.; Islam, Rafique; Tong, Fan; Ghavami, Maryam; Mutunga, James M.; Slebodnick, Carla; Li, Jianyong; Viayna, Elisabet; Lam, Polo C.-H.; Totrov, Maxim M.; Bloomquist, Jeffrey R.; Carlier, Paul R.

    2015-01-01

    To identify potential selective and resistance-breaking mosquitocides against the African malaria vector Anopheles gambiae, we investigated the acetylcholinesterase (AChE) inhibitory and mosquitocidal properties of isoxazol-3-yl dimethylcarbamates (15), and the corresponding 3-oxoisoxazole-2(3H)-dimethylcarboxamide isomers (14). In both series, compounds were found with excellent contact toxicity to wild-type susceptible (G3) strain and multiply resistant (Akron) strain mosquitoes that carry the G119S resistance mutation of AChE. Compounds possessing good to excellent toxicity to Akron strain mosquitoes inhibit the G119S mutant of An. gambiae AChE (AgAChE) with ki values at least 10- to 600-fold higher than that of propoxur, a compound that does not kill Akron mosquitoes at the highest concentration tested. On average, inactivation of WT AgAChE by dimethylcarboxamides 14 was 10-20 fold faster than that of the corresponding isoxazol-3-yl dimethylcarbamates 15. X-ray crystallography of dimethylcarboxamide 14d provided insight into that reactivity, a finding that may explain the inhibitory power of structurally-related inhibitors of hormone-sensitive lipase. Finally, human/An. gambiae AChE inhibition selectivities of these compounds were low, suggesting the need for additional structural modification. PMID:25684426

  20. A spatial individual-based model predicting a great impact of copious sugar sources and resting sites on survival of Anopheles gambiae and malaria parasite transmission

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    Zhu, Lin; Qualls, Whitney A.; Marshall, John M; Arheart, Kris L.; DeAngelis, Don; McManus, John W.; Traore, Sekou F.; Doumbia, Seydou; Schlein, Yosef; Muller, Gunter C.; Beier, John C.

    2015-01-01

    BackgroundAgent-based modelling (ABM) has been used to simulate mosquito life cycles and to evaluate vector control applications. However, most models lack sugar-feeding and resting behaviours or are based on mathematical equations lacking individual level randomness and spatial components of mosquito life. Here, a spatial individual-based model (IBM) incorporating sugar-feeding and resting behaviours of the malaria vector Anopheles gambiae was developed to estimate the impact of environmental sugar sources and resting sites on survival and biting behaviour.MethodsA spatial IBM containing An. gambiae mosquitoes and humans, as well as the village environment of houses, sugar sources, resting sites and larval habitat sites was developed. Anopheles gambiae behaviour rules were attributed at each step of the IBM: resting, host seeking, sugar feeding and breeding. Each step represented one second of time, and each simulation was set to run for 60 days and repeated 50 times. Scenarios of different densities and spatial distributions of sugar sources and outdoor resting sites were simulated and compared.ResultsWhen the number of natural sugar sources was increased from 0 to 100 while the number of resting sites was held constant, mean daily survival rate increased from 2.5% to 85.1% for males and from 2.5% to 94.5% for females, mean human biting rate increased from 0 to 0.94 bites per human per day, and mean daily abundance increased from 1 to 477 for males and from 1 to 1,428 for females. When the number of outdoor resting sites was increased from 0 to 50 while the number of sugar sources was held constant, mean daily survival rate increased from 77.3% to 84.3% for males and from 86.7% to 93.9% for females, mean human biting rate increased from 0 to 0.52 bites per human per day, and mean daily abundance increased from 62 to 349 for males and from 257 to 1120 for females. All increases were significant (P Survival was greater when sugar sources were randomly distributed

  1. The Cry4B toxin of Bacillus thuringiensis subsp. israelensis kills Permethrin-resistant Anopheles gambiae, the principal vector of malaria.

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    Ibrahim, Mohamed A; Griko, Natalya B; Bulla, Lee A

    2013-04-01

    Resurgence of malaria has been attributed, in part, to the development of resistance by Anopheles gambiae, a principal vector of the disease, to various insecticidal compounds such as Permethrin. Permethrin, a neurotoxicant, is widely used to impregnate mosquito nets. An alternative strategy to control mosquitoes is the use of Bacillus thuringiensis subsp. israelensis (Bti) because there is no observable resistance in the field to the bacterium. Bti kills mosquitoes by targeting cadherin molecules residing in the midgut epithelium of larvae of the insect. Cry proteins (Cry4A, Cry4B, Cry10A and Cry11A) produced by the bacterium during the sporulation phase of its life cycle bind to the cadherin molecules, which serve as receptors for the proteins. These Cry proteins have variable specificity to a variety of mosquitoes, including Culex and Aedes as well as Anopheles. Importantly, selective mosquitocidal action is occasioned by binding of the respective Cry toxins to cadherins distinctive to individual mosquito species. Differential fractionation of the four Cry proteins from a novel Bti isolate (M1) and cloning and expression of their genes in Escherichia coli revealed that Cry4B is the only Cry protein that exerts insecticidal action against An. gambiae. Indeed, it does so against a Permethrin-resistant strain of the mosquito. The other three Cry proteins are ineffective. Multiple sequence alignments of the four Cry proteins revealed a divergent sequence motif in the Cry4B toxin, which most likely determines binding of the toxin to its cognate receptor, BT-R3, in An. gambiae and to its specific toxicity. A model showing Cry4B toxin binding to BT-R3 is presented. PMID:23760000

  2. Anopheles darlingi and Anopheles marajoara (Diptera: Culicidae susceptibility to pyrethroids in an endemic area of the Brazilian Amazon

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    Allan Kardec Ribeiro Galardo

    2015-12-01

    Full Text Available Abstract: INTRODUCTION: This study aimed to evaluate the susceptibility of Anopheles darlingi Root (1926 and Anopheles marajoara Galvão & Damasceno (1942 to pyrethroids used by the National Malaria Control Program in Brazil. METHODS: Mosquitoes from Amapá, Brazilian Amazon, were assessed for resistance to cypermethrin, deltamethrin, and alpha-cypermethrin. Insecticide-impregnated bottles were used as suggested by the CDC/Atlanta. RESULTS: Diagnostic dose for Anopheles darlingi was 12.5µg/bottle during 30 min of exposure. Concentrations for Anopheles marajoara were 20µg/bottle of cypermethrin and deltamethrin and 12.5µg/bottle of alpha-cypermethrin. CONCLUSIONS : No resistance was recorded for Anopheles darlingi , but Anopheles marajoara requires attention.

  3. Sequencing and analysis of the complete mitochondrial genome in Anopheles culicifacies species B (Diptera: Culicidae).

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    Hua, Ya-Qiong; Yan, Zhen-Tian; Fu, Wen-Bo; He, Qi-Yi; Zhou, Yong; Chen, Bin

    2016-07-01

    The complete mitochondrial genome sequence of Anopheles culicifacial species B was sequenced in this study. The length of the mitochondrial genome is 15 330 bp, which contains 13 protein-coding genes (PCGs), 22 tRNA genes, two rRNA genes, and a non-coding control region. The gene order and the gene composition are consistent with those previously reported for other mosquito species. The initiation codon of the PCGs complies with the ATN rule except for COI using TCG and ND5 using GTG as a start codon, and the termination codon is TAA or imcomplete, an only T. The total base composition is 40.4% A, 38.1% T, 12.4% C, and 9.1% G. The phylogenetic tree based on the sequences of 13 protein-coding genes showed that these species were classified into two clades, corresponding to the subgenus Cellia and subgenus Nyssorhynchus. An. culicifacies species B of Myzomyia Series was clustered with An. gambiae of Pyretophorus Series with a high bootstrap value of 100%. The complete mitogenome data can provide a basis for molecular identification and phylogenetic studies of mosquito species. PMID:26114319

  4. Olyset Duo® (a pyriproxyfen and permethrin mixture net: an experimental hut trial against pyrethroid resistant Anopheles gambiae and Culex quinquefasciatus in Southern Benin.

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    Corine Ngufor

    Full Text Available BACKGROUND: Alternative compounds which can complement pyrethroids on long-lasting insecticidal nets (LN in the control of pyrethroid resistant malaria vectors are urgently needed. Pyriproxyfen (PPF, an insect growth regulator, reduces the fecundity and fertility of adult female mosquitoes. LNs containing a mixture of pyriproxyfen and pyrethroid could provide personal protection through the pyrethroid component and reduce vector abundance in the next generation through the sterilizing effect of pyriproxyfen. METHOD: The efficacy of Olyset Duo, a newly developed mixture LN containing pyriproxyfen and permethrin, was evaluated in experimental huts in southern Benin against pyrethroid resistant Anopheles gambiae and Culex quinquefasciatus. Comparison was made with Olyset Net® (permethrin alone and a LN with pyriproxyfen alone (PPF LN. Laboratory tunnel tests were performed to substantiate the findings in the experimental huts. RESULTS: Overall mortality of wild pyrethroid resistant An. gambiae s.s. was significantly higher with Olyset Duo than with Olyset Net (50% vs. 27%, P = 0.01. Olyset DUO was more protective than Olyset Net (71% vs. 3%, P<0.001. The oviposition rate of surviving blood-fed An. gambiae from the control hut was 37% whereas none of those from Olyset Duo and PPF LN huts laid eggs. The tunnel test results were consistent with the experimental hut results. Olyset Duo was more protective than Olyset Net in the huts against wild pyrethroid resistant Cx. quinquefasciatus although mortality rates of this species did not differ significantly between Olyset Net and Olyset Duo. There was no sterilizing effect on surviving blood-fed Cx. quinquefasciatus with the PPF-treated nets. CONCLUSION: Olyset Duo was superior to Olyset Net in terms of personal protection and killing of pyrethroid resistant An. gambiae, and sterilized surviving blood-fed mosquitoes. Mixing pyrethroid and pyriproxyfen on a LN shows potential for malaria control and

  5. Molecular characterization and evolution of a gene family encoding male-specific reproductive proteins in the African malaria vector Anopheles gambiae

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    Sharakhov Igor V

    2011-10-01

    Full Text Available Abstract Background During copulation, the major Afro-tropical malaria vector Anopheles gambiae s.s. transfers male accessory gland (MAG proteins to females as a solid mass (i.e. the "mating plug". These proteins are postulated to function as important modulators of female post-mating responses. To understand the role of selective forces underlying the evolution of these proteins in the A. gambiae complex, we carried out an evolutionary analysis of gene sequence and expression divergence on a pair of paralog genes called AgAcp34A-1 and AgAcp34A-2. These encode MAG-specific proteins which, based on homology with Drosophila, have been hypothesized to play a role in sperm viability and function. Results Genetic analysis of 6 species of the A. gambiae complex revealed the existence of a third paralog (68-78% of identity, that we named AgAcp34A-3. FISH assays showed that this gene maps in the same division (34A of chromosome-3R as the other two paralogs. In particular, immuno-fluorescence assays targeting the C-terminals of AgAcp34A-2 and AgAcp34A-3 revealed that these two proteins are localized in the posterior part of the MAG and concentrated at the apical portion of the mating plug. When transferred to females, this part of the plug lies in proximity to the duct connecting the spermatheca to the uterus, suggesting a potential role for these proteins in regulating sperm motility. AgAcp34A-3 is more polymorphic than the other two paralogs, possibly because of relaxation of purifying selection. Since both unequal crossing-over and gene conversion likely homogenized the members of this gene family, the interpretation of the evolutionary patterns is not straightforward. Although several haplotypes of the three paralogs are shared by most A. gambiae s.l. species, some fixed species-specific replacements (mainly placed in the N- and C-terminal portions of the secreted peptides were also observed, suggesting some lineage-specific adaptation. Conclusions

  6. Distribution of ace-1R and resistance to carbamates and organophosphates in Anopheles gambiae s.s. populations from Côte d'Ivoire

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    Ahoua Alou Ludovic P

    2010-06-01

    Full Text Available Abstract Background The spread of pyrethroid resistance in Anopheles gambiae s.s. is a critical issue for malaria vector control based on the use of insecticide-treated nets. Carbamates and organophosphates insecticides are regarded as alternatives or supplements to pyrethroids used in nets treatment. It is, therefore, essential to investigate on the susceptibility of pyrethroid resistant populations of An. gambiae s.s. to these alternative products. Methods In September 2004, a cross sectional survey was conducted in six localities in Côte d'Ivoire: Toumbokro, Yamoussoukro, Toumodi in the Southern Guinea savannah, Tiassalé in semi-deciduous forest, then Nieky and Abidjan in evergreen forest area. An. gambiae populations from these localities were previously reported to be highly resistant to pyrethroids insecticides. Anopheline larvae were collected from the field and reared to adults. Resistance/susceptibility to carbamates (0.4% carbosulfan, 0.1% propoxur and organophosphates (0.4% chlorpyrifos-methyl, 1% fenitrothion was assessed using WHO bioassay test kits for adult mosquitoes. Then, PCR assays were run to determine the molecular forms (M and (S, as well as phenotypes for insensitive acetylcholinesterase (AChE1 due to G119S mutation. Results Bioassays showed carbamates (carbosulfan and propoxur resistance in all tested populations of An. gambiae s.s. In addition, two out of the six tested populations (Toumodi and Tiassalé were also resistant to organophosphates (mortality rates ranged from 29.5% to 93.3%. The M-form was predominant in tested samples (91.8%. M and S molecular forms were sympatric at two localities but no M/S hybrids were detected. The highest proportion of S-form (7.9% of An. gambiae identified was in sample from Toumbokro, in the southern Guinea savannah. The G119S mutation was found in both M and S molecular forms with frequency from 30.9 to 35.2%. Conclusion This study revealed a wide distribution of insensitive

  7. Mosquito nets treated with a mixture of chlorfenapyr and alphacypermethrin control pyrethroid resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes in West Africa.

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    Raphael N'Guessan

    Full Text Available BACKGROUND: The effectiveness of insecticide treated nets is under threat across Africa south of the Sahara from the selection of pyrethroid resistance in Anopheles gambiae mosquitoes. To maintain progress against malaria it is necessary to identify alternative residual insecticides for mosquito nets. Mixtures of pyrethroid and insecticides with novel mode of action provide scope for both improved control and management of resistance through concurrent exposure to unrelated insecticides. METHODS: The pyrrole chlorfenapyr and the pyrethroid alphacypermethrin were tested individually and as a mixture on mosquito nets in an experimental hut trial in southern Benin against pyrethroid resistant An gambiae and Culex quinquefasciatus mosquitoes. The nets were deliberately holed to simulate the effect of wear and tear. RESULTS: The nets treated with the mixture of chlorfenapyr 200 mg/m² and alphacypermethrin 25 mg/m² killed a proportion of An gambiae (77%, 95%CI: 66-86% significantly greater than nets treated with alphacypermethrin 25 mg/m(2 (30%, 95%CI: 21-41% but not significantly different from nets treated with chlorfenapyr 200 mg/m² (69%, 95%CI: 57-78%. The nets treated with the mixtures procured personal protection against An gambiae biting(58-62% by a greater margin than the alphacypermethrin treated net (39%, whereas the chlorfenapyr treated net was not protective. A similar trend in mortality and blood feeding inhibition between treatments was observed in Cx quinquefasciatus to that seen in An. gambiae, although the effects were lower. A mixture of alphacypermethrin with chlorfenapyr applied at 100 mg/m² had an effect similar to the mixture with chlorfenapyr at 200 mg/m². CONCLUSION: The effectiveness of ITNs against pyrethroid resistant mosquitoes was restored by the mixture: the alphacypermethrin component reduced human-vector contact while the chlorfenapyr controlled pyrethroid-resistant mosquitoes. The complementary action of these

  8. Impact of three years of large scale Indoor Residual Spraying (IRS and Insecticide Treated Nets (ITNs interventions on insecticide resistance in Anopheles gambiae s.l. in Benin

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    Padonou Gil

    2012-04-01

    Full Text Available Abstract Background In Benin, Indoor Residual Spraying (IRS and long-lasting insecticidal nets (LLINs are the cornerstones of malaria prevention. In the context of high resistance of Anopheles gambiae to pyrethroids, The National Malaria Control Program (NMCP has undertaken a full coverage of IRS in a no-flood zone in the Oueme region, coupled with the distribution of LLINs in a flood zone. We assessed the impact of this campaign on phenotypic resistance, kdr (knock-down resistance and ace-1R (insensitive acetylcholinesterase mutations. Methods Insecticides used for malaria vector control interventions were bendiocarb WP (0.4 g/m2 and deltamethrin (55 mg/m2, respectively for IRS and LLINs. Susceptibility status of An. gambiae was assessed using World Health Organization bioassay tests to DDT, permethrin, deltamethrin and bendiocarb in the Oueme region before intervention (2007 and after interventions in 2008 and 2010. An. gambiae specimens were screened for identification of species, molecular M and S forms and for the detection of the West African kdr (L1014F as well as ace-1R mutations using PCR techniques. Results The univariate logistic regression performed showed that kdr frequency has increased significantly during the three years in the intervention area and in the control area. Several factors (LLINs, IRS, mosquito coils, aerosols, use of pesticides for crop protection could explain the selection of individual resistant An. gambiae. The Kdr resistance gene could not be the only mechanism of resistance observed in the Oueme region. The high susceptibility to bendiocarb is in agreement with a previous study conducted in Benin. However, the occurrence of ace-1R heterozygous individuals even on sites far from IRS areas, suggests other factors may contribute to the selection of resistance other than those exerted by the vector control program. Conclusion The results of this study have confirmed that An.gambiae have maintained and developed

  9. [Laboratory evaluation of alpha-cypermethrin insecticide efficacy on Anopheles gambiae populations of Côte d'Ivoire resistant to permethrin and deltamethrin].

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    Koffi, A A; Darriet, F; N'Guessan, R; Doannio, J M; Carnevale, P

    1999-02-01

    Susceptibility tests were carried out in laboratory conditions to evaluate the efficacy of alpha-cypermethrin (a synthetic pyrethroid never used in Côte d'Ivoire) for malaria vector control. Five wild populations of Anopheles gambiae originating from M'bé, Yaokoffikro, Korhogo, Kafiné and Daola and two laboratory reared strains (Kisumu susceptible and Kou permethrin resistant selected strain) were tested. The diagnostic dosage of alpha-cypermethrin for the sensitive strain Kisumu was 2.5 x 10(-3)%. A comparative study of the susceptibility of samples of wild populations of An. gambiae was carried out according to the WHO standard susceptibility test. Impregnated papers with 4% DDT, 0.25% permethrin, 0.025% deltamethrin and 0.0025% alpha-cypermethrin were used. The results showed that except for mosquitoes from M'bé, all the other populations were resistant to these insecticides. Bioassays were carried out with alpha-cypermethrin at the operational dosage of 20 mg a.i./m2 on the same population and laboratory reared strains. The results showed the efficacy of this insecticide on both the Kisumu strain and the population from M'bé, a maintained efficacy for the Daloa, Kafiné and Korhogo mosquito populations, but the wild anopheline population from Yaokoffikro clearly appeared fully resistant.

  10. Combining two-dimensional gel electrophoresis and metabolomic data in support of dry-season survival in the two main species of the malarial mosquito Anopheles gambiae

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    Hidalgo K.

    2015-12-01

    Full Text Available In dry savannahs of West-Africa, the malarial mosquitoes of the Anopheles gambiae sensu stricto complex annually survive the harsh desiccating conditions of the dry season. However, the physiological and biochemical mechanisms underlying how these mosquitoes survive such desiccating conditions are still undefined, and controversial. In this context, we provide the first work examining both proteomic and metabolomic changes in the two molecular forms of A. gambiae s.s (M and S forms experimentally exposed to the rainy and dry season conditions as they experience in the field. Protein abundances of the mosquitoes were measured using a two-dimensional fluorescence difference gel electrophoresis (2D DIGE coupled with a matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF and tandem mass spectrometry (MS for protein identification. These assays were conducted by Applied Biomics (http://www.appliedbiomics.com, Applied Biomics, Inc. Hayward, CA, USA, and the mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org via the PRIDE partner repository with the dataset identifier PXD000294. The metabolomic analysis was conducted using both Acquity UPLC® system (for amino acid identification, and a gas-chromatography-mass spectrometry platform (for sugars identification. Metabolomic fingerprintings were assessed in the University of Rennes 1, UMR CNRS 6553 EcoBio (France. A detailed interpretation of the obtained data can be found in Hidalgo et al. (2014 [1] (Journal of Insect Physiology (2014.

  11. Effects of co-habitation between Anopheles gambiae s.s. and Culex quinquefasciatus aquatic stages on life history traits

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    Kweka Eliningaya J

    2012-02-01

    Full Text Available Abstract Background The effective measures for the control of malaria and filariasis vectors can be achieved by targeting immature stages of anopheline and culicine mosquitoes in productive habitat. To design this strategy, the mechanisms (like biotic interactions with conspecifc and heterospecific larvae regulating mosquito aquatic stages survivorship, development time and the size of emerging adults should be understood. This study explored the effect of co-habitation between An. gambiae s.s. and Cx. quinquefasciatus on different life history traits of both species under different densities and constant food supply in the habitats of the same size under semi-natural conditions. Methods Experiments were set up with three combinations; Cx. quinquefasciatus alone (single species treatment, An. gambiae s.s. alone (single species treatment; and An. gambiae s.s. with Cx. quiquefasciatus (co-habitation treatment in different densities in semi field situation. Results The effect of co-habitation of An. gambiae s.s. and Cx. quinquefasciatus was found to principally affect three parameters. The wing-lengths (a proxy measure of body size of An. gambiae s.s. in co-habitation treatments were significantly shorter in both females and males than in An. gambiae s.s single species treatments. In Cx. quinquefasciatus, no significant differences in wing-length were observed between the single species and co-habitation treatments. Daily survival rates were not significantly different between co-habitation and single species treatments for both An. gambiae s.s. and Cx. quinquefasciatus. Developmental time was found to be significantly different with single species treatments developing better than co-habitation treatments. Sex ratio was found to be significantly different from the proportion of 0.5 among single and co-habitation treatments species at different densities. Single species treatments had more males than females emerging while in co

  12. Expression of trypsin modulating oostatic factor (TMOF in an entomopathogenic fungus increases its virulence towards Anopheles gambiae and reduces fecundity in the target mosquito

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    Kamareddine Layla

    2013-01-01

    Full Text Available Abstract Background Adult and larval mosquitoes regulate food digestion in their gut with trypsin modulating oostatic factor (TMOF, a decapeptide hormone synthesized by the ovaries and the neuroendocrine system. TMOF is currently being developed as a mosquitocide, however, delivery of the peptide to the mosquito remains a significant challenge. Entomopathogenic fungi offer a means for targeting mosquitoes with TMOF. Findings The efficacy of wild type and transgenic Beauveria bassiana strains expressing Aedes aegypti TMOF (Bb-Aa1 were evaluated against larvae and sugar- and blood-fed adult Anopheles gambiae mosquitoes using insect bioassays. Bb-Aa1 displayed increased virulence against larvae, and sugar and blood fed adult A. gambiae when compared to the wild type parent strain. Median lethal dose (LD50 values decreased by ~20% for larvae, and ~40% for both sugar and blood-fed mosquitoes using Bb-Aa1 relative to the wild type parent. Median lethal time (LT50 values were lower for blood-fed compared to sugar-fed mosquitoes in infections with both wild type and Bb-Aa1. However, infection using Bb-Aa1 resulted in 15% to 25% reduction in LT50 values for sugar- and blood fed mosquitoes, and ~27% for larvae, respectively, relative to the wild type parent. In addition, infection with Bb-Aa1 resulted in a dramatic reduction in fecundity of the target mosquitoes. Conclusions B. bassiana expressing Ae. aegypti TMOF exhibited increased virulence against A. gambiae compared to the wild type strain. These data expand the range and utility of entomopathogenic fungi expressing mosquito-specific molecules to improve their biological control activities against mosquito vectors of disease.

  13. Transcriptome profiling of chemosensory appendages in the malaria vector Anopheles gambiae reveals tissue- and sex-specific signatures of odor coding

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    Rokas Antonis

    2011-05-01

    Full Text Available Abstract Background Chemosensory signal transduction guides the behavior of many insects, including Anopheles gambiae, the major vector for human malaria in sub-Saharan Africa. To better understand the molecular basis of mosquito chemosensation we have used whole transcriptome RNA sequencing (RNA-seq to compare transcript expression profiles between the two major chemosensory tissues, the antennae and maxillary palps, of adult female and male An. gambiae. Results We compared chemosensory tissue transcriptomes to whole body transcriptomes of each sex to identify chemosensory enhanced genes. In the six data sets analyzed, we detected expression of nearly all known chemosensory genes and found them to be highly enriched in both olfactory tissues of males and females. While the maxillary palps of both sexes demonstrated strict chemosensory gene expression overlap, we observed acute differences in sensory specialization between male and female antennae. The relatively high expression levels of chemosensory genes in the female antennae reveal its role as an organ predominately assigned to chemosensation. Remarkably, the expression of these genes was highly conserved in the male antennae, but at much lower relative levels. Alternatively, consistent with a role in mating, the male antennae displayed significant enhancement of genes involved in audition, while the female enhancement of these genes was observed, but to a lesser degree. Conclusions These findings suggest that the chemoreceptive spectrum, as defined by gene expression profiles, is largely similar in female and male An. gambiae. However, assuming sensory receptor expression levels are correlated with sensitivity in each case, we posit that male and female antennae are perceptive to the same stimuli, but possess inverse receptive prioritizations and sensitivities. Here we have demonstrated the use of RNA-seq to characterize the sensory specializations of an important disease vector and

  14. Transcriptional mediators Kto and Skd are involved in the regulation of the IMD pathway and anti-Plasmodium defense in Anopheles gambiae.

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    Yang Chen

    Full Text Available The malarial parasite Plasmodium must complete a complex lifecycle in its Anopheles mosquito host, the main vector for Plasmodium. The mosquito resists infection with the human malarial parasite P. falciparum by engaging the NF-κB immune signaling pathway, IMD. Here we show that the conserved transcriptional mediators Kto and Skd are involved in the regulation of the mosquito IMD pathway. RNAi-mediated depletion of Kto and Skd in the Anopheles gambiae cell line L5-3 resulted in a decrease in the transcript abundance of Cec1, which is controlled by the IMD pathway. Silencing the two genes also resulted in an increased susceptibility of the mosquito to bacterial and Plasmodium falciparum infection, but not to infection with the rodent malaria parasite P. berghei. We also showed that Kto and Skd are not transcriptional co-activators of Rel2 or other key factors of the IMD pathway; however, they participate in the regulation of the IMD pathway, which is crucial for the mosquito's defense against P. falciparum.

  15. Successful field trial of attractive toxic sugar bait (ATSB plant-spraying methods against malaria vectors in the Anopheles gambiae complex in Mali, West Africa

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    Doumbia Seydou

    2010-07-01

    Full Text Available Abstract Background Based on highly successful demonstrations in Israel that attractive toxic sugar bait (ATSB methods can decimate local populations of mosquitoes, this study determined the effectiveness of ATSB methods for malaria vector control in the semi-arid Bandiagara District of Mali, West Africa. Methods Control and treatment sites, selected along a road that connects villages, contained man-made ponds that were the primary larval habitats of Anopheles gambiae and Anopheles arabiensis. Guava and honey melons, two local fruits shown to be attractive to An. gambiae s.l., were used to prepare solutions of Attractive Sugar Bait (ASB and ATSB that additionally contained boric acid as an oral insecticide. Both included a color dye marker to facilitate determination of mosquitoes feeding on the solutions. The trial was conducted over a 38-day period, using CDC light traps to monitor mosquito populations. On day 8, ASB solution in the control site and ATSB solution in the treatment site were sprayed using a hand-pump on patches of vegetation. Samples of female mosquitoes were age-graded to determine the impact of ATSB treatment on vector longevity. Results Immediately after spraying ATSB in the treatment site, the relative abundance of female and male An. gambiae s.l. declined about 90% from pre-treatment levels and remained low. In the treatment site, most females remaining after ATSB treatment had not completed a single gonotrophic cycle, and only 6% had completed three or more gonotrophic cycles compared with 37% pre-treatment. In the control site sprayed with ASB (without toxin, the proportion of females completing three or more gonotrophic cycles increased from 28.5% pre-treatment to 47.5% post-treatment. In the control site, detection of dye marker in over half of the females and males provided direct evidence that the mosquitoes were feeding on the sprayed solutions. Conclusion This study in Mali shows that even a single application of

  16. Activité larvicide sur Anopheles gambiae Giles et composition chimique des huiles essentielles extraites de quatre plantes cultivées au Cameroun

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    Tchoumbougnang F.

    2009-01-01

    Full Text Available Larvicidal activity against Anopheles gambiae Giles and chemical composition of essential oils from four plants cultivated in Cameroon. The chemical composition of the essential oils obtained by hydrodistillation of dry leaves from Cymbopogon citrates (DC. Stapf, Ocimum canum Sims, Ocimum gratissimum L. var 'gratissimum' L. and Thymus vulgaris L. cultivated in Cameroon were analyzed and their larvicidal activity against fourth instar larvae of Anopheles gambiae Giles were determined. The yields of extraction indicate that T. vulgaris (0.95% was richer in essential oil than C. citratus (0.67%. O. canum and O. gratissimum have approximately the same content in volatile constituents (0.59% and 0.60%, respectively. The analyses by GC and GC/MS showed that these oils are monoterpenic (86.8-97.4%. Oxygenated monoterpenes predominate in C. citratus and O. canum (81.6% and 68.9%, respectively while O. gratissimum oil contains a majority of monoterpene hydrocarbons (61.0%. T. vulgaris is characterized by the same proportion of monoterpene hydrocarbons (45.6% and oxygenated monoterpenes (48.9%. The main compounds found in the essential oil of C. citratus are acyclic monoterpenes such as geraniol (15.6%, geranial (39.3%, neral (21.9% and myrcene (14.0%. The essential oil of T. vulgaris is characterized by p-menthane structures given by thymol (40.1%, p-cymene (23.4% and γ-terpinene (15.1%; p-cymene (32.1% and thymol (24.3% were also the constituents quantitatively important in O. gratissimum essential oil while linalool (56.3% and limonene (10.9% were predominant in O. canum. Bioassay test done by the World Health Organization standard protocol revealed that these essential oils have remarkable larvicidal properties as they could induce 100% mortality in the larvae of A. gambiae at the concentration of 100 ppm for C. citratus, 200 ppm with T. vulgaris, 350 ppm for O. gratissimum and 400 ppm for O. canum. Their LC50 and LC80 show the same reactivity order

  17. Humoral response to the Anopheles gambiae salivary protein gSG6: a serological indicator of exposure to Afrotropical malaria vectors.

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    Cinzia Rizzo

    Full Text Available Salivary proteins injected by blood feeding arthropods into their hosts evoke a saliva-specific humoral response which can be useful to evaluate exposure to bites of disease vectors. However, saliva of hematophagous arthropods is a complex cocktail of bioactive factors and its use in immunoassays can be misleading because of potential cross-reactivity to other antigens. Toward the development of a serological marker of exposure to Afrotropical malaria vectors we expressed the Anopheles gambiae gSG6, a small anopheline-specific salivary protein, and we measured the anti-gSG6 IgG response in individuals from a malaria hyperendemic area of Burkina Faso, West Africa. The gSG6 protein was immunogenic and anti-gSG6 IgG levels and/or prevalence increased in exposed individuals during the malaria transmission/rainy season. Moreover, this response dropped during the intervening low transmission/dry season, suggesting it is sensitive enough to detect variation in vector density. Members of the Fulani ethnic group showed higher anti-gSG6 IgG response as compared to Mossi, a result consistent with the stronger immune reactivity reported in this group. Remarkably, anti-gSG6 IgG levels among responders were high in children and gradually declined with age. This unusual pattern, opposite to the one observed with Plasmodium antigens, is compatible with a progressive desensitization to mosquito saliva and may be linked to the continued exposure to bites of anopheline mosquitoes. Overall, the humoral anti-gSG6 IgG response appears a reliable serological indicator of exposure to bites of the main African malaria vectors (An. gambiae, Anopheles arabiensis and, possibly, Anopheles funestus and it may be exploited for malaria epidemiological studies, development of risk maps and evaluation of anti-vector measures. In addition, the gSG6 protein may represent a powerful model system to get a deeper understanding of molecular and cellular mechanisms underlying the

  18. Trends in DDT and pyrethroid resistance in Anopheles gambiae s.s. populations from urban and agro-industrial settings in southern Cameroon

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    Kerah-Hinzoumbé Clément

    2009-09-01

    Full Text Available Abstract Background Pyrethroid insecticides are widely used for insect pest control in Cameroon. In certain insect species, particularly the malaria vector Anopheles gambiae, resistance to this class of insecticides is a source of great concern and needs to be monitored in order to sustain the efficacy of vector control operations in the fields. This study highlights trends in DDT and pyrethroid resistance in wild An. gambiae populations from South Cameroon. Methods Mosquitoes were collected between 2001 and 2007 in four sites in South Cameroon, where insecticides are used for agricultural or personal protection purposes. Insecticide use was documented in each site by interviewing residents. Batches of 2-4 days old adult female mosquitoes reared from larval collections were tested for susceptibility to DDT, permethrin and deltamethrin using standard WHO procedures. Control, dead and survivors mosquitoes from bioassays were identified by PCR-RFLP and characterized for the kdr mutations using either the AS-PCR or the HOLA method. Results Four chemical insecticide groups were cited in the study sites: organochlorines, organophosphates, carbamates and pyrethroids. These chemicals were used for personal, crop or wood protection. In the four An. gambiae populations tested, significant variation in resistance levels, molecular forms composition and kdr frequencies were recorded in the time span of the study. Increases in DDT and pyrethroid resistance, as observed in most areas, were generally associated with an increase in the relative frequency of the S molecular form carrying the kdr mutations at higher frequencies. In Mangoum, however, where only the S form was present, a significant increase in the frequency of kdr alleles between 2003 to 2007 diverged with a decrease of the level of resistance to DDT and pyrethroids. Analyses of the kdr frequencies in dead and surviving mosquitoes showed partial correlation between the kdr genotypes and resistance

  19. Bionomics of Anopheles spp. (Diptera: Culicidae) in a malaria endemic region of Sukabumi, West Java, Indonesia.

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    Stoops, Craig A; Rusmiarto, Saptoro; Susapto, Dwiko; Munif, Amurl; Andris, Heri; Barbara, Kathryn A; Sukowati, Supratman

    2009-12-01

    A 15-month bionomic study of Anopheles species was conducted in two ecologically distinct villages (coastal and upland) of Sukabumi District, West Java, Indonesia from June 2006 to September 2007. Mosquitoes were captured using human-landing collections at both sites. During the study, a total of 17,100 Anopheles mosquitoes comprising 13 Anopheles species were caught: 9,151 at the coastal site and 7,949 at the upland site. Anopheles barbirostris, Anopheles maculatus, and Anopheles vagus were the predominant species caught at the coastal site, and Anopheles aconitus, Anopheles barbirostris, and An. maculatus predominated in the upland site. Overall, species were exophagic at both sites, but there was variation between species. Anopheles aconitus was endophagic at the coastal site, exophagic at the upland site, collected most often in April 2007 and had a peak landing time between 22:00 and 23:00. Anopheles sundaicus was only collected at the coastal site, exophagic, collected most often in October 2006, and had a peak landing time between 19:00 and 20:00. Potential malaria vector species such An. aconitus, An. maculatus, and An. sundaicus were present throughout the year. None of the 7,770 Anopheles tested using CSP-ELISA were positive for malaria, although the risk for malaria outbreaks in Sukabumi district remains high.

  20. The Anopheles albitarsis complex with the recognition of Anopheles oryzalimnetes Wilkerson and Motoki, n. sp. and Anopheles janconnae Wilkerson and Sallum, n. sp. (Diptera: Culicidae

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    Maysa Tiemi Motoki

    2009-09-01

    Full Text Available The Anopheles (Nyssorhynchus albitarsis complex includes six species: An. albitarsis, Anopheles oryzalimnetes Wilkerson and Motoki, n. sp., Anopheles marajoara, Anopheles deaneorum, Anopheles janconnae Wilkerson and Sallum, n. sp. and An. albitarsis F. Except for An. deaneorum, species of the complex are indistinguishable when only using morphology. The problematic distinction among species of the complex has made study of malaria transmission and ecology of An. albitarsis s.l. difficult. Consequently, involvement of species of the An. albitarsis complex in human Plasmodium transmission is not clear throughout its distribution range. With the aim of clarifying the taxonomy of the above species, with the exception of An. albitarsis F, we present comparative morphological and morphometric analyses, morphological redescriptions of three species and description of two new species using individuals from populations in Brazil, Paraguay, Argentina and Venezuela. The study included characters from adult females, males, fourth-instar larvae, pupae and male genitalia of An. albitarsis, An. marajoara, An. deaneorum and An. oryzalimnetes n. sp. For An. janconnae n. sp. only characters of the female, male and male genitalia were analyzed. Fourth-instar larvae, pupae and male genitalia characteristics of all five species are illustrated. Bionomics and distribution data are given based on published literature records.

  1. Crystal and solution studies of the "Plus-C" odorant-binding protein 48 from Anopheles gambiae: control of binding specificity through three-dimensional domain swapping.

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    Tsitsanou, Katerina E; Drakou, Christina E; Thireou, Trias; Vitlin Gruber, Anna; Kythreoti, Georgia; Azem, Abdussalam; Fessas, Dimitrios; Eliopoulos, Elias; Iatrou, Kostas; Zographos, Spyros E

    2013-11-15

    Much physiological and behavioral evidence has been provided suggesting that insect odorant-binding proteins (OBPs) are indispensable for odorant recognition and thus are appealing targets for structure-based discovery and design of novel host-seeking disruptors. Despite the fact that more than 60 putative OBP-encoding genes have been identified in the malaria vector Anopheles gambiae, the crystal structures of only six of them are known. It is therefore clear that OBP structure determination constitutes the bottleneck for structure-based approaches to mosquito repellent/attractant discovery. Here, we describe the three-dimensional structure of an A. gambiae "Plus-C" group OBP (AgamOBP48), which exhibits the second highest expression levels in female antennae. This structure represents the first example of a three-dimensional domain-swapped dimer in dipteran species. A combined binding site is formed at the dimer interface by equal contribution of each monomer. Structural comparisons with the monomeric AgamOBP47 revealed that the major structural difference between the two Plus-C proteins localizes in their N- and C-terminal regions, and their concerted conformational change may account for monomer-swapped dimer conversion and furthermore the formation of novel binding pockets. Using a combination of gel filtration chromatography, differential scanning calorimetry, and analytical ultracentrifugation, we demonstrate the AgamOBP48 dimerization in solution. Eventually, molecular modeling calculations were used to predict the binding mode of the most potent synthetic ligand of AgamOBP48 known so far, discovered by ligand- and structure-based virtual screening. The structure-aided identification of multiple OBP binders represents a powerful tool to be employed in the effort to control transmission of the vector-borne diseases. PMID:24097978

  2. Evaluation of two counterflow traps for testing behaviour-mediating compounds for the malaria vector Anopheles gambiae s.s. under semi-field conditions in Tanzania

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    Killeen Gerry F

    2008-11-01

    Full Text Available Abstract Background Evaluation of mosquito responses towards different trap-bait combinations in field trials is a time-consuming process that can be shortened by experiments in contained semi-field systems. Possible use of the BG Sentinel (BGS trap to sample Anopheles gambiae s.s. was evaluated. The efficiency of this trap was compared with that of the Mosquito Magnet-X (MM-X trap, when baited with foot odour alone or combinations of foot odour with carbon dioxide (CO2 or lemongrass as behaviour-modifying cues. Methods Female An. gambiae s.s. were released in an experimental flight arena that was placed in a semi-field system and left overnight. Catch rates for the MM-X and BGS traps were recorded. Data were analysed by fitting a generalized linear model to the (n+1 transformed catches. Results Both types of traps successfully captured mosquitoes with all odour cues used. When the BGS trap was tested against the MM-X trap in a choice assay with foot odour as bait, the BGS trap caught about three times as many mosquitoes as the MM-X trap (P = 0.002. Adding CO2 (500 ml/min to foot odour increased the number of mosquitoes caught by 268% for the MM-X (P Conclusion The BGS trap shows high potential for field trials due to its simple construction and high catch rate when baited with human foot odour only. However, for rapid screening of different baits in a contained semi-field system, the superior discriminatory power of the MM-X trap is advantageous.

  3. Effects of co-habitation between Anopheles gambiae s.s. and Culex quinquefasciatus aquatic stages on life history traits

    OpenAIRE

    Kweka Eliningaya J; Zhou Goufa; Beilhe Leila B; Dixit Amruta; Afrane Yaw; Gilbreath Thomas M; Munga Stephen; Nyindo Mramba; Githeko Andrew K; Yan Guiyun

    2012-01-01

    Abstract Background The effective measures for the control of malaria and filariasis vectors can be achieved by targeting immature stages of anopheline and culicine mosquitoes in productive habitat. To design this strategy, the mechanisms (like biotic interactions with conspecifc and heterospecific larvae) regulating mosquito aquatic stages survivorship, development time and the size of emerging adults should be understood. This study explored the effect of co-habitation between An. gambiae s...

  4. The Anopheles gambiae Oxidation Resistance 1 (OXR1) Gene Regulates Expression of Enzymes That Detoxify Reactive Oxygen Species

    OpenAIRE

    Jaramillo-Gutierrez, Giovanna; Molina-Cruz, Alvaro; Kumar, Sanjeev; Barillas-Mury, Carolina

    2010-01-01

    Background OXR1 is an ancient gene, present in all eukaryotes examined so far that confers protection from oxidative stress by an unknown mechanism. The most highly conserved region of the gene is the carboxyl-terminal TLDc domain, which has been shown to be sufficient to prevent oxidative damage. Methodology/Principal Findings OXR1 has a complex genomic structure in the mosquito A. gambiae, and we confirm that multiple splice forms are expressed in adult females. Our studies revealed that OX...

  5. Clarification of anomalies in the application of a 2La molecular karyotyping method for the malaria vector Anopheles gambiae

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    Knols Bart GJ

    2008-12-01

    Full Text Available Abstract Background Chromosomal inversions have been considered to be potentially important barriers to gene flow in many groups of animals through their effect on recombination suppression in heterokaryotypic individuals. Inversions can also enhance local adaptation in different groups of organisms and may often represent species-specific differences among closely related taxa. We conducted a study to characterize the 2La inversion karyotypes of An. gambiae sensu stricto mosquitoes sampled from the Kilombero Valley (Tanzania using a newly designed PCR assay. Results We frequently encountered a (687 bp fragment which was only present in the Kilombero Valley populations. Laboratory crossing between An. gambiae s.s. from Njage (Tanzania and Kisumu (Western Kenya populations resulted in F1 offspring carrying the observed fragment. Karyotype analysis did not indicate differences in 2La region chromosome morphology between individuals carrying the PCR fragments, the 207 bp fragment, or the 687 bp fragement. Conclusion The observed insertion/deletion polymorphism within the region amplified by the 2La PCR diagnostic test may confound the interpretation of this assay and should be well considered in order to maintain an acceptable level of reliability in studies using this assay to describe the distribution and frequency of the 2La inversion among natural populations of An. gambiae s.s.

  6. A Qualitative Evidence of the Breeding Sites of Anopheles arabiensis Patton (Diptera: Culicidae) in and Around Kassala Town, Eastern Sudan.

    Science.gov (United States)

    Hamza, Asma Mahmoud; El Rayah, El Amin

    2016-01-01

    Anopheles arabiensis Patton (Diptera: Culicidae) is considered the most efficient malaria vector in eastern Sudan. This study aims to characterize the breeding sites of An. arabiensis throughout the year in and around Kassala town, eastern Sudan. Diverse larval habitat types were visited and characterized based on the habitat type and chemical composition. Mosquito larvae were found in many diverse habitats. During the rainy season, rain pools and water bodies created by the seasonal Gash River serve as the main breeding sites. In the dry season, irrigation canals, seepage from water pipes, neglected wells, artificial containers, and man-made ditches serve as the main breeding sites. Breeding water showed a pH of 7.9 and a low concentration of the total dissolved salts. The results of this study may be considered in planning and implementing larval control programs in the area. PMID:27547039

  7. A Possible Mechanism for the Suppression of Plasmodium berghei Development in the Mosquito Anopheles gambiae by the Microsporidian Vavraia culicis

    OpenAIRE

    Bargielowski, Irka; Koella, Jacob C

    2009-01-01

    Background Microsporidian parasites of mosquitoes offer a possible way of controlling malaria, as they impede the development of Plasmodium parasites within the mosquito. The mechanism involved in this interference process is unknown. Methodology We evaluated the possibility that larval infection by a microsporidian primes the immune system of adult mosquitoes in a way that enables a more effective anti-Plasmodium response. To do so, we infected 2-day old larvae of the mosquito Anopheles gamb...

  8. [Resistance of Anopheles gambiae s.l. to pyrethroids and DDT at Tiassalékro, an irrigated rice-growing village in the southern forest of Ivory Coast].

    Science.gov (United States)

    Konan, K G; Koné, A B; Konan, Y L; Fofana, D; Konan, K L; Diallo, A; Ziogba, J C; Touré, M; Kouassi, K P; Doannio, J M C

    2011-10-01

    An assessment of the sensitivity of Anopheles gambiae s.l.to three pyrethroids (alphacypermethrin, permethrin, deltamethrin) and DDT has been carried out with a laboratory strain (Kisumu reference sensitive strain) and a wild strain (Tiassalékro strain) using larvae from an irrigated rice-growing area of Tiassalékro, located in the southern forest of Ivory Coast. The sensitivity tests were performed according to the standard WHO cylinder tests with adult female A. gambiae s.l. aged 2 to 4 days. The results showed that the strain of Tiassalékro is resistant to the three tested pyrethroids and DDT. The molecular forms M and S were identified, with a predominance of M form. The resistance mechanism involved is the Kdr mutation. In this region, control measures against malaria vectors by using bed nets impregnated with these insecticides or household sprays could be compromised.

  9. Mosquitoes of Anopheles hyrcanus (Diptera, Culicidae) Group: Species Diagnostic and Phylogenetic Relationships.

    Science.gov (United States)

    Khrabrova, Natalia V; Andreeva, Yulia V; Sibataev, Anuarbek K; Alekseeva, Svetlana S; Esenbekova, Perizat A

    2015-09-01

    Herein, we report the results of study of Anopheles species in Primorsk and Khabarovsk regions of Russia. Three species of the Anopheles hyrcanus group: An. kleini, An. pullus, and An. lesteri were identified by molecular taxonomic diagnostics for the first time in Russia. Surprisingly, An. sinensis, which earlier was considered the only species of Anopheles in Russian Far East, was not observed. We analyzed nucleotide variation in the 610-bp fragment of the 5' end of the cytochrome c oxidase subunit I (COI) region. All species possessed a distinctive set of COI sequences. A maximum likelihood phylogenetic tree was constructed for members of the hyrcanus group. The examined Anopheles hyrcanus group members could be divided into two major subgroups: subgroup 1 (An. hyrcanus and An. pullus) and subgroup 2 (An. sinensis, An. kleini, and An. lesteri), which were found to be monophyletic.

  10. Molecular comparison of topotypic specimens confirms Anopheles (Nyssorhynchus dunhami Causey (Diptera: Culicidae in the Colombian Amazon

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    Freddy Ruiz

    2010-11-01

    Full Text Available The presence of Anopheles (Nyssorhynchus dunhami Causey in Colombia (Department of Amazonas is confirmed for the first time through direct comparison of mtDNA cytochrome c oxidase I (COI barcodes and nuclear rDNA second internal transcribed spacer (ITS2 sequences with topotypic specimens of An. dunhami from Tefé, Brazil. An. dunhami was identified through retrospective correlation of DNA sequences following misidentification as Anopheles nuneztovari s.l. using available morphological keys for Colombian mosquitoes. That An. dunhami occurs in Colombia and also possibly throughout the Amazon Basin, is of importance to vector control programs, as this non-vector species is morphologically similar to known malaria vectors including An. nuneztovari, Anopheles oswaldoi and Anopheles trinkae. Species identification of An. dunhami and differentiation from these closely related species are highly robust using either DNA ITS2 sequences or COI DNA barcode. DNA methods are advocated for future differentiation of these often sympatric taxa in South America.

  11. Interaction affinity of Delta and Epsilon class glutathione-s-transferases (GSTs to bind with DDT for detoxification and conferring resistance in Anopheles gambiae, a malaria vector

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    V. Aravindan

    2014-01-01

    Full Text Available Background & objectives: The enzyme glutathione-s-transferases (GSTs are associated with detoxification of DDT, as experimentally proved in Anopheles gambiae. Insect GSTs are classified into six classes and among them Delta and Epsilon class GSTs have been implicated in detoxification of organochlorine insecticides. Both Delta and Epsilon GSTs produce, in total, 24 transcripts that result in the production of corresponding enzyme proteins. However, the conventional assay estimates the level of total GSTs and relates to development of resistance to DDT. Hence, it would be more reliable to estimate the level of the specific class GSTs that shows higher affinity with DDT. This would also lead to design a specific molecular tool for resistance diagnosis. Methods: Of the 24 GSTs, computational models for 23 GSTs, which are available in Swiss-Prot database, were retrieved and for the remaining one, D7-2, for which no model is available in the data bank, a structural model was developed using the sequence of An. dirus B with a PDB ID of 1R5A as the template. All the models were docked with DDT in the presence of reduced glutathione. Results: The energy output showed that Delta, D6 has the highest interaction affinity with DDT. Hence, this particular GST (D6 is likely to get elevated on exposure of mosquitoes to DDT. Interpretation & conclusion: It would be, therefore, possible to design a specific molecular assay to determine the expression level of such high affinity transcript(s and to use for resistance diagnosis reliably in the vector surveillance programme.

  12. Anopheles gambiae odorant binding protein crystal complex with the synthetic repellent DEET: implications for structure-based design of novel mosquito repellents.

    Science.gov (United States)

    Tsitsanou, K E; Thireou, T; Drakou, C E; Koussis, K; Keramioti, M V; Leonidas, D D; Eliopoulos, E; Iatrou, K; Zographos, S E

    2012-01-01

    Insect odorant binding proteins (OBPs) are the first components of the olfactory system to encounter and bind attractant and repellent odors emanating from various sources for presentation to olfactory receptors, which trigger relevant signal transduction cascades culminating in specific physiological and behavioral responses. For disease vectors, particularly hematophagous mosquitoes, repellents represent important defenses against parasitic diseases because they effect a reduction in the rate of contact between the vectors and humans. OBPs are targets for structure-based rational approaches for the discovery of new repellent or other olfaction inhibitory compounds with desirable features. Thus, a study was conducted to characterize the high resolution crystal structure of an OBP of Anopheles gambiae, the African malaria mosquito vector, in complex with N,N-diethyl-m-toluamide (DEET), one of the most effective repellents that has been in worldwide use for six decades. We found that DEET binds at the edge of a long hydrophobic tunnel by exploiting numerous non-polar interactions and one hydrogen bond, which is perceived to be critical for DEET's recognition. Based on the experimentally determined affinity of AgamOBP1 for DEET (K (d) of 31.3 μΜ) and our structural data, we modeled the interactions for this protein with 29 promising leads reported in the literature to have significant repellent activities, and carried out fluorescence binding studies with four highly ranked ligands. Our experimental results confirmed the modeling predictions indicating that structure-based modeling could facilitate the design of novel repellents with enhanced binding affinity and selectivity. PMID:21671117

  13. Structure-function analysis of the Anopheles gambiae LRIM1/APL1C complex and its interaction with complement C3-like protein TEP1.

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    Michael Povelones

    2011-04-01

    Full Text Available Malaria threatens half the world's population and exacts a devastating human toll. The principal malaria vector in Africa, the mosquito Anopheles gambiae, encodes 24 members of a recently identified family of leucine-rich repeat proteins named LRIMs. Two members of this family, LRIM1 and APL1C, are crucial components of the mosquito complement-like pathway that is important for immune defense against Plasmodium parasites. LRIM1 and APL1C circulate in the hemolymph exclusively as a disulfide-bonded complex that specifically interacts with the mature form of the complement C3-like protein, TEP1. We have investigated the specificity of LRIM1/APL1C complex formation and which regions of these proteins are required for interactions with TEP1. To address these questions, we have generated a set of LRIM1 and APL1C alleles altering key conserved structural elements and assayed them in cell culture for complex formation and interaction with TEP1. Our data indicate that heterocomplex formation is an intrinsic ability of LRIM1 and APL1C and identify key homologous cysteine residues forming the intermolecular disulfide bond. We also demonstrate that the coiled-coil domain is the binding site for TEP1 but also contributes to the specificity of LRIM1/APL1C complex formation. In addition, we show that the LRIM1/APL1C complex interacts with the mature forms of three other TEP proteins, one of which, TEP3, we have characterized as a Plasmodium antagonist. We conclude that LRIM1 and APL1C contain three distinct modules: a C-terminal coiled-coil domain that can carry different TEP protein cargoes, potentially with distinct functions, a central cysteine-rich region that controls complex formation and an N-terminal leucine-rich repeat with a putative role in pathogen recognition.

  14. Anopheles gambiae distribution and insecticide resistance in the cities of Douala and Yaoundé (Cameroon: influence of urban agriculture and pollution

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    Awono-Ambene Parfait

    2011-06-01

    Full Text Available Abstract Background Urban malaria is becoming a major health priority across Africa. A study was undertaken to assess the importance of urban pollution and agriculture practice on the distribution and susceptibility to insecticide of malaria vectors in the two main cities in Cameroon. Methods Anopheline larval breeding sites were surveyed and water samples analysed monthly from October 2009 to December 2010. Parameters analysed included turbidity, pH, temperature, conductivity, sulfates, phosphates, nitrates, nitrites, ammonia, aluminium, alkalinity, iron, potassium, manganese, magnesium, magnesium hardness and total hardness. Characteristics of water bodies in urban areas were compared to rural areas and between urban sites. The level of susceptibility of Anopheles gambiae to 4% DDT, 0.75% permethrin, 0.05% deltamethrin, 0.1% bendiocarb and 5% malathion were compared between mosquitoes collected from polluted, non polluted and cultivated areas. Results A total of 1,546 breeding sites, 690 in Yaoundé and 856 in Douala, were sampled in the course of the study. Almost all measured parameters had a concentration of 2- to 100-fold higher in urban compare to rural breeding sites. No resistance to malathion was detected, but bendiocarb resistance was present in Yaounde. Very low mortality rates were observed following DDT or permethrin exposure, associated with high kdr frequencies. Mosquitoes collected in cultivated areas, exhibited the highest resistant levels. There was little difference in insecticide resistance or kdr allele frequency in mosquitoes collected from polluted versus non-polluted sites. Conclusion The data confirm high selection pressure on mosquitoes originating from urban areas and suggest urban agriculture rather than pollution as the major factor driving resistance to insecticide.

  15. Increase in susceptibility to insecticides with aging of wild Anopheles gambiae mosquitoes from Côte d’Ivoire

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    Chouaibou Mouhamadou S

    2012-09-01

    Full Text Available Abstract Background Appropriate monitoring of vector insecticide susceptibility is required to provide the rationale for optimal insecticide selection in vector control programs. Methods In order to assess the influence of mosquito age on susceptibility to various insecticides, field-collected larvae of An. gambiae s.l. from Tiassalé were reared to adults. Females aged 1, 2, 3, 5 and 10 days were exposed to 5 insecticides (deltamethrin, permethrin, DDT, malathion and propoxur using WHO susceptibility test kits. Outcome measures included the LT50 (exposure time required to achieve 50% knockdown, the RR (resistance ratio, i.e. a calculation of how much more resistant the wild population is compared with a standard susceptible strain and the mortality rate following 1 hour exposure, for each insecticide and each mosquito age group. Results There was a positive correlation between the rate of knockdown and mortality for all the age groups and for all insecticides tested. For deltamethrin, the RR50 was highest for 2 day old and lowest for 10 day old individuals. Overall, mortality was lowest for 2 and 3 day old individuals and significantly higher for 10 day old individuals (P 50 was highest for 1 to 3 day old individuals and lowest for 10 day old individuals and mortality was lowest for 1 to 3 day old individuals, intermediate for 5 day old and highest for 10 day old individuals. DDT did not display any knockdown effect and mortality was low for all mosquito age groups (50 was low (1.54 - 2.77 and mortality was high (>93% for all age groups. With propoxur, no knockdown effect was observed for 1, 2 and 3 day old individuals and a very low level of mortality was observed ( Conclusion Results indicate that for An. gambiae s.l. adults derived from wild-collected larvae, there was an influence of age on insecticide susceptibility status, with younger individuals (1 to 3 days old more resistant than older mosquitoes. This

  16. Role of Anopheles (Kerteszia bellator as malaria vector in Southeastern Brazil (Diptera: Culicidae

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    Oswaldo Paulo Forattini

    1999-11-01

    Full Text Available New research concerning Anopheles bellator in the southeast of the State of São Paulo, Brazil, are reported. Adult females of this mosquito showed remarkable endophily and endophagy which was even greater than An. cruzii. The epidemiological role of this anopheline as a malaria vector is discussed.

  17. Detection of knockdown resistance (kdr mutations in Anopheles gambiae: a comparison of two new high-throughput assays with existing methods

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    Ball Amanda

    2007-08-01

    Full Text Available Abstract Background Knockdown resistance (kdr is a well-characterized mechanism of resistance to pyrethroid insecticides in many insect species and is caused by point mutations of the pyrethroid target site the para-type sodium channel. The presence of kdr mutations in Anopheles gambiae, the most important malaria vector in Africa, has been monitored using a variety of molecular techniques. However, there are few reports comparing the performance of these different assays. In this study, two new high-throughput assays were developed and compared with four established techniques. Methods Fluorescence-based assays based on 1 TaqMan probes and 2 high resolution melt (HRM analysis were developed to detect kdr alleles in An. gambiae. Four previously reported techniques for kdr detection, Allele Specific Polymerase Chain Reaction (AS-PCR, Heated Oligonucleotide Ligation Assay (HOLA, Sequence Specific Oligonucleotide Probe – Enzyme-Linked ImmunoSorbent Assay (SSOP-ELISA and PCR-Dot Blot were also optimized. The sensitivity and specificity of all six assays was then compared in a blind genotyping trial of 96 single insect samples that included a variety of kdr genotypes and African Anopheline species. The relative merits of each assay was assessed based on the performance in the genotyping trial, the length/difficulty of each protocol, cost (both capital outlay and consumable cost, and safety (requirement for hazardous chemicals. Results The real-time TaqMan assay was both the most sensitive (with the lowest number of failed reactions and the most specific (with the lowest number of incorrect scores. Adapting the TaqMan assay to use a PCR machine and endpoint measurement with a fluorimeter showed a slight reduction in sensitivity and specificity. HRM initially gave promising results but was more sensitive to both DNA quality and quantity and consequently showed a higher rate of failure and incorrect scores. The sensitivity and specificity of AS

  18. Chitosan/DsiRNA nanoparticle targeting identifies AgCad1 cadherin in Anopheles gambiae larvae as an in vivo receptor of Cry11Ba toxin of Bacillus thuringiensis subsp. jegathesan.

    Science.gov (United States)

    Zhang, Qi; Hua, Gang; Adang, Michael J

    2015-05-01

    The Cry11Ba protein of Bacillus thuringiensis subsp. jegathesan crystals has uniquely high toxicity against a spectrum of mosquito species. The high potency of Cry11Ba against Anopheles gambiae is caused by recognition of multiple midgut proteins including glycosyl phosphatidylinositol-anchored alkaline phosphatase AgALP1, aminopeptidase AgAPN2, α-amylase AgAmy1 and α-glucosidase Agm3 that bind Cry11Ba with high affinity and function as putative receptors. The cadherin AgCad2 in An. gambiae larvae also binds Cry11Ba with high affinity (Kd = 12 nM) and is considered a putative receptor, while cadherin AgCad1 bound Cry11Ba with low affinity (Kd = 766 nM), a property not supportive for a Cry11Ba receptor role. Here, we show the in vivo involvement of AgCad1 in Cry11Ba toxicity in An. gambiae larvae using chitosan/DsiRNA nanoparticles to inhibit AgCad expression in larvae. Cry11Ba was significantly less toxic to AgCad1-silenced larvae than to control larvae. Because AgCad1 was co-suppressed by AgCad2 DsRNAi, the involvement of AgCad2 in Cry11Ba toxicity could not be ascertained. The ratio of AgCad1:AgCad2 transcript level is 36:1 for gut tissue in 4th instar larvae. Silencing AgCad expression had no effect on transcript levels of other binding receptors of Cry11Ba. We conclude that AgCad1 and possibly AgCad2 in An. gambiae larvae are functional receptors of Cry11Ba toxin in vivo. PMID:25758367

  19. Mosquito repellent potential of Pithecellobium dulce leaf and seed against malaria vector Anopheles stephensi (Diptera: Culicidae)

    OpenAIRE

    Mohan Rajeswary; Marimuthu Govindarajan

    2016-01-01

    Objective: To determine the repellent properties of hexane, benzene, ethyl acetate, chloroform and methanol extract of Pithecellobium dulce (P. dulce) leaf and seed against Anopheles stephensi (An. stephensi). Methods: Repellent activity assay was carried out in a net cage (45 cm × 30 cm × 25 cm) containing 100 blood starved female mosquitoes of An. stephensi. This assay was carried out in the laboratory conditions according to the WHO 2009 protocol. Plant crude extracts of P. ...

  20. Contrasted Fitness Costs of Docking and Antibacterial Constructs in the EE and EVida3 Strains Validates Two-Phase Anopheles gambiae Genetic Transformation System.

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    Doug Paton

    Full Text Available The deployment of transgenic mosquitoes carrying genes for refractoriness to malaria has long been seen as a futuristic scenario riddled with technical difficulties. The integration of anti-malarial effector genes and a gene-drive system into the mosquito genome without affecting mosquito fitness is recognized as critical to the success of this malaria control strategy. Here we conducted detailed fitness studies of two Anopheles gambiae s.s. transgenic lines recently developed using a two-phase targeted genetic transformation system. In replicated cage-invasion experiments, males and females of the EE Phase-1 docking strain and EVida3 Phase-2 strain loaded with an antimicrobial peptide (AMP expressed upon blood-feeding, were mixed with individuals of a recently-colonized strain of the Mopti chromosomal form. The experimental design enabled us to detect initial strain reproductive success differences, assortative mating and hybrid vigor that may characterize mosquito release situations. In addition, the potential fitness costs of the unloaded Phase-1 and loaded Phase-2 genetic constructs, independent of the strains' original genetic backgrounds, were estimated between the 1(st instar larvae, pupae and adult stages over 10 generations. The Phase-1 unloaded docking cassette was found to have significantly lower allelic fitness relative to the wild type allele during larval development. However, overall genotypic fitness was comparable to the wild type allele across all stages leading to stable equilibrium in all replicates. In contrast, the Phase-2 construct expressing EVida3 disappeared from all replicates within 10 generations due to lower fitness of hemi- and homozygous larvae, suggesting costly background AMP expression and/or of the DsRed2 marker. This is the first study to effectively partition independent fitness stage-specific determinants in unloaded and loaded transgenic strains of a Phase-1-2 transformation system. Critically, the high

  1. Physiological correlates of ecological divergence along an urbanization gradient: differential tolerance to ammonia among molecular forms of the malaria mosquito Anopheles gambiae

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    Tene Fossog Billy

    2013-01-01

    Full Text Available Abstract Background Limitations in the ability of organisms to tolerate environmental stressors affect their fundamental ecological niche and constrain their distribution to specific habitats. Evolution of tolerance, therefore, can engender ecological niche dynamics. Forest populations of the afro-tropical malaria mosquito Anopheles gambiae have been shown to adapt to historically unsuitable larval habitats polluted with decaying organic matter that are found in densely populated urban agglomerates of Cameroon. This process has resulted in niche expansion from rural to urban environments that is associated with cryptic speciation and ecological divergence of two evolutionarily significant units within this taxon, the molecular forms M and S, among which reproductive isolation is significant but still incomplete. Habitat segregation between the two forms results in a mosaic distribution of clinally parapatric patches, with the M form predominating in the centre of urban agglomerates and the S form in the surrounding rural localities. We hypothesized that development of tolerance to nitrogenous pollutants derived from the decomposition of organic matter, among which ammonia is the most toxic to aquatic organisms, may affect this pattern of distribution and process of niche expansion by the M form. Results Acute toxicity bioassays indicated that populations of the two molecular forms occurring at the extremes of an urbanization gradient in Yaounde, the capital of Cameroon, differed in their response to ammonia. The regression lines best describing the dose-mortality profile differed in the scale of the explanatory variable (ammonia concentration log-transformed for the S form and linear for the M form, and in slope (steeper for the S form and shallower for the M form. These features reflected differences in the frequency distribution of individual tolerance thresholds in the two populations as assessed by probit analysis, with the M form exhibiting

  2. Contrasted Fitness Costs of Docking and Antibacterial Constructs in the EE and EVida3 Strains Validates Two-Phase Anopheles gambiae Genetic Transformation System.

    Science.gov (United States)

    Paton, Doug; Underhill, Anne; Meredith, Janet; Eggleston, Paul; Tripet, Frederic

    2013-01-01

    The deployment of transgenic mosquitoes carrying genes for refractoriness to malaria has long been seen as a futuristic scenario riddled with technical difficulties. The integration of anti-malarial effector genes and a gene-drive system into the mosquito genome without affecting mosquito fitness is recognized as critical to the success of this malaria control strategy. Here we conducted detailed fitness studies of two Anopheles gambiae s.s. transgenic lines recently developed using a two-phase targeted genetic transformation system. In replicated cage-invasion experiments, males and females of the EE Phase-1 docking strain and EVida3 Phase-2 strain loaded with an antimicrobial peptide (AMP) expressed upon blood-feeding, were mixed with individuals of a recently-colonized strain of the Mopti chromosomal form. The experimental design enabled us to detect initial strain reproductive success differences, assortative mating and hybrid vigor that may characterize mosquito release situations. In addition, the potential fitness costs of the unloaded Phase-1 and loaded Phase-2 genetic constructs, independent of the strains' original genetic backgrounds, were estimated between the 1(st) instar larvae, pupae and adult stages over 10 generations. The Phase-1 unloaded docking cassette was found to have significantly lower allelic fitness relative to the wild type allele during larval development. However, overall genotypic fitness was comparable to the wild type allele across all stages leading to stable equilibrium in all replicates. In contrast, the Phase-2 construct expressing EVida3 disappeared from all replicates within 10 generations due to lower fitness of hemi- and homozygous larvae, suggesting costly background AMP expression and/or of the DsRed2 marker. This is the first study to effectively partition independent fitness stage-specific determinants in unloaded and loaded transgenic strains of a Phase-1-2 transformation system. Critically, the high fitness of the

  3. Evaluation of leaf aqueous extract and synthesized silver nanoparticles using Nerium oleander against Anopheles stephensi (Diptera: Culicidae).

    Science.gov (United States)

    Roni, Mathath; Murugan, Kadarkarai; Panneerselvam, Chellasamy; Subramaniam, Jayapal; Hwang, Jiang-Shiou

    2013-03-01

    Green nanoparticle synthesis has been achieved using environmentally acceptable plant extract and ecofriendly reducing and capping agents. The present study was carried out to establish the larvicidal activity of synthesized silver nanoparticles (AgNPs) using leaf extract of Nerium oleander (Apocynaceae) against the first to fourth instar larvae and pupae of malaria vector, Anopheles stephensi (Diptera: Culicidae). Nanoparticles are being used in many commercial applications. It was found that aqueous silver ions can be reduced by the aqueous extract of the plant parts to generate extremely stable silver nanoparticles in water. The results were recorded from UV-Vis spectrum, Fourier transform infrared (FTIR) spectroscopy, scanning electron microscopy (SEM), and energy-dispersive X-ray (EDX) spectroscopy analysis. The production of the AgNPs synthesized using leaf extract of N. oleander was evaluated through a UV-Vis spectrophotometer in a wavelength range of 200 to 700 nm. This revealed a peak at 440 nm in N. oleander leaf extracts, indicating the production of AgNPs. The FTIR spectra of AgNPs exhibited prominent peaks at 509.12 cm(-1) (C-H bend alkenes), 1,077.05 cm(-1) (C-O stretch alcohols), 1,600.63 cm(-1) (N-H bend amines), 2,736.49 and 2,479.04 cm(-1) (O-H stretch carboxylic acids), and 3,415.31 cm(-1) (N-H stretching due to amines group). An SEM micrograph showed 20-35-nm-size aggregates of spherical- and cubic-shaped nanoparticles. EDX showed the complete chemical composition of the synthesized nanoparticles of silver. Larvicidal activity of aqueous leaf extract of N. oleander and synthesized AgNPs was carried out against Anopheles stephensi, and the results showed that the highest larval mortality was found in the synthesized AgNPs against the first to fourth instar larvae and pupae of Anopheles stephensi with the following values: LC(50) of instar larvae 20.60, 24.90, 28.22, and 33.99 ppm; LC(90) of instar larvae 41.62, 50.33, 57.78, and 68.41

  4. Expression of the cytochrome P450s, CYP6P3 and CYP6M2 are significantly elevated in multiple pyrethroid resistant populations of Anopheles gambiae s.s. from Southern Benin and Nigeria

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    Ranson Hilary

    2008-11-01

    Full Text Available Abstract Background Insecticide resistance in Anopheles mosquitoes is threatening the success of malaria control programmes. This is particularly true in Benin where pyrethroid resistance has been linked to the failure of insecticide treated bed nets. The role of mutations in the insecticide target sites in conferring resistance has been clearly established. In this study, the contribution of other potential resistance mechanisms was investigated in Anopheles gambiae s.s. from a number of localities in Southern Benin and Nigeria. The mosquitoes were sampled from a variety of breeding sites in a preliminary attempt to investigate the role of contamination of mosquito breeding sites in selecting for resistance in adult mosquitoes. Results All mosquitoes sampled belonged to the M form of An. gambiae s.s. There were high levels of permethrin resistance in an agricultural area (Akron and an urban area (Gbedjromede, low levels of resistance in mosquito samples from an oil contaminated site (Ojoo and complete susceptibility in the rural Orogun location. The target site mutation kdrW was detected at high levels in two of the populations (Akron f = 0.86 and Gbedjromede f = 0.84 but was not detected in Ojoo or Orogun. Microarray analysis using the Anopheles gambiae detox chip identified two P450s, CYP6P3 and CYP6M2 up regulated in all three populations, the former was expressed at particularly high levels in the Akron (12.4-fold and Ojoo (7.4-fold populations compared to the susceptible population. Additional detoxification and redox genes were also over expressed in one or more populations including two cuticular pre-cursor genes which were elevated in two of the three resistant populations. Conclusion Multiple resistance mechanisms incurred in the different breeding sites contribute to resistance to permethrin in Benin. The cytochrome P450 genes, CYP6P3 and CYP6M2 are upregulated in all three resistant populations analysed. Several additional potential

  5. Bioefficacy of Morinda tinctoria and Pongamia glabra plant extracts against the malaria vector Anopheles stephensi (Diptera: Culicidae

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    D. Amerasan

    2015-04-01

    Full Text Available Mosquito-borne diseases have an economic impact, including loss in commercial and labour outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. The aim of the present study was to investigate the larvicidal, adulticidal and ovicidal activity of dried leaf chloroform, ethyl acetate, acetone, aqueous, and methanol extracts of Morinda tinctoria and Pongamia glabra against larvae of Anopheles stephensi (Diptera: Culicidae. Larvae were exposed to varying concentrations of plant extracts for 24 h. All extracts showed moderate larvicidal effects after 24 h of exposure; however, the highest larval mortality was found with the leaf methanol extracts of M. tinctoria and P. glabra against the larvae of A. stephensi lethal concentration (LC50=136.24 and 141.05 ppm; LC90=342.67 and 368.89 ppm, respectively. The results of the adulticidal activity assays of chloroform, ethyl acetate, acetone, aqueous, and methanol extracts of M. tinctoria and P. glabra showed significant mortality against larvae of A. stephensi. The methanol extract showed maximum activity compared with the other extracts. The greatest effect on mean percentage hatch in the ovicidal assays was observed 48 h post-treatment. Percent hatch was inversely proportional to the concentration of extract, and directly proportional to the number of eggs. A mortality of 100% was observed with 100-400 ppm methanol extracts and 200-400 ppm aqueous extracts of M. tinctoria, and 200-400 ppm aqueous and methanol extracts of P. glabra. This study provides the first report of the larvicidal, adulticidal and ovicidal activities of M. tinctoria and P. glabra plant extracts against the malaria vector, A. stephensi, representing an ideal eco-friendly approach for its control.

  6. Random amplified polymorphic DNA analysis of Anopheles nuneztovari (Diptera: Culicidae from Western and Northeastern Colombia

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    Carmen Elisa Posso

    2003-06-01

    Full Text Available Random amplified polymorphic DNA (RAPD markers were used to analyze 119 DNA samples of three Colombian Anopheles nuneztovari populations to study genetic variation and structure. Genetic diversity, estimated from heterozygosity, averaged 0.34. Genetic flow was greater between the two populations located in Western Colombia (F ST: 0.035; Nm: 6.8 but lower between these two and the northeastern population (F ST: 0.08; Nm: 2.8. According to molecular variance analysis, the genetic distance between populations was significant (phiST 0.1131, P < 0.001. The variation among individuals within populations (phiST 0.8869, P < 0.001was also significant, suggesting a greater degree of population subdivision, not considered in this study. Both the parameters evaluated and the genetic flow suggest that Colombian An. nuneztovari populations are co-specific.

  7. Cytogenetic evidence for a species complex within Anopheles pseudopunctipennis theobald (Diptera: Culicidae).

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    Coetzee, M; Estrada-Franco, J G; Wunderlich, C A; Hunt, R H

    1999-04-01

    Anopheles pseudopunctipennis was collected from Acapulco, Mexico and Sallee River, Grenada, West Indies and used in cross-mating experiments. Larvae from the cross, Mexico female X Grenada male, died in the third instar. However, adult progeny were obtained from the reciprocal cross Grenada female x Mexico male. These hybrid males had testes with apparently normal appearance but some without viable sperm. Polytene chromosomes obtained from hybrid females exhibited extensive asynapsis of the X chromosomes. Previously undescribed fixed inversion differences between the two populations were noted on the X chromosome. It is concluded that the two populations belong to different species. The Grenada population is designated An. pseudopunctipennis species C, since it is the third taxon recognized in this species complex.

  8. Systematic studies on Anopheles galvaoi Causey, Deane & Deane from the subgenus Nysssorhynchus blanchard (Diptera: Culicidae

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    Maria Anice Mureb Sallum

    2002-12-01

    Full Text Available Anopheles galvaoi, a member of the subgenus Nyssorhynchus, is redescribed based on morphological characters of the adults male and female, fourth-instar larva and pupa. Female, male genitalia, larval and pupal stages are illustrated. Data about medical importance, bionomics, and distribution are given based on literature records. Adult female of An. galvaoi can be easily misidentified as An. benarrochi Gabaldón and An. aquasalis Curry. A few characters are indicated for identifying female and immatures of An. galvaoi. Phylogenetic relationships among An. galvaoi and six other species of the Oswaldoi Subgroup are estimated using COII mtDNA and ITS2 rDNA gene sequences. Lectotype of An. galvaoi, an adult female from Rio Branco, State of Acre, is invalidated.

  9. [Historical review of the distribution of Anopheles (Nyssorhynchus) darlingi (Diptera: Culicidae) in the Peruvian Amazon].

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    Fernández, Roberto; Vera, Hubert; Calderón, Guillermo

    2014-04-01

    Anopheles (Nyssorhynchus) darlingi has been reported since 1931 in border areas of the department of Loreto, mainly along the borders with Brazil and Colombia. In 1994, during an outbreak of malaria, An. darlingi was found in neighboring towns to Iquitos. At present, its distribution has expanded considerably in Loreto. This paper reviews literature available for all possible information on the distribution of mosquitoes, particularly anopheline in the Amazon region of the country, with special emphasis on An darlingi. Entomological collections were also conducted in the departments of Madre de Dios and Ucayali in order to know and verify the distribution of An. darlingi. At present, the distribution of the species is confined to localities in southeastern Peru with Bolivia border towns, in a town near the Abujao River in the department of Ucayali, and widely in the northeastern region of the Amazon basin of Loreto in Peru. PMID:25123872

  10. Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strategies for malaria vector control in Africa using attractive toxic sugar bait methods

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    Bah Sekou

    2010-09-01

    Full Text Available Abstract Background Based on recent studies in Israel demonstrating that attractive toxic sugar bait (ATSB methods can be used to decimate local anopheline and culicine mosquito populations, an important consideration is whether the same methods can be adapted and improved to attract and kill malaria vectors in Africa. The ATSB approach uses fruit or flower scent as an attractant, sugar solution as a feeding stimulant, and an oral toxin. The ATSB solutions are either sprayed on vegetation or suspended in simple bait stations, and the mosquitoes ingesting the toxic solutions are killed. As such, this approach targets sugar-feeding female and male mosquitoes. This study examines the attractiveness of African malaria vectors to local fruits/seedpods and flowering plants, key biological elements of the ATSB approach for mosquito control. Methods Three field experiments were conducted at sites in Mali. The attraction of Anopheles gambiae s.l. to 26 different local fruits and seedpods was determined at a site in the semi-arid Bandiagara District of Mali. Wire mesh glue traps with fruits/seedpods suspended on skewers inside were set along a seasonal lagoon. Seven replicates of each fruit/seedpod species were tested, with a water-soaked sponge and a sugar-soaked sponge as controls. The attraction of An. gambiae s.l. to 26 different types of flowering plants was determined at a site near Mopti in Mali. The flowering plants held in a water-filled buried container were tested using the same glue traps, with controls including water only and sugar solution. Six replicates of each selected plant type were tested on transects between rice paddies. Additional studies using CDC light traps were done to determine the relative densities and periodicity of An. gambiae s.l. attraction to branches of the most highly attractive flowering plant, branches without flowers, human odor, and candescent light. Results Of the 26 fruits and seedpods tested, 6 were attractive

  11. Molecular taxonomy of Anopheles (Nyssorhynchus) benarrochi (Diptera: Culicidae) and malaria epidemiology in southern Amazonian Peru.

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    Conn, Jan E; Moreno, Marta; Saavedra, Marlon; Bickersmith, Sara A; Knoll, Elisabeth; Fernandez, Roberto; Vera, Hubert; Burrus, Roxanne G; Lescano, Andres G; Sanchez, Juan Francisco; Rivera, Esteban; Vinetz, Joseph M

    2013-02-01

    Anopheline specimens were collected in 2011 by human landing catch, Shannon and CDC traps from the malaria endemic localities of Santa Rosa and San Pedro in Madre de Dios Department, Peru. Most specimens were either Anopheles (Nyssorhynchus) benarrochi B or An. (Nys.) rangeli, confirmed by polymerase chain reaction-restriction fragment length polymorphism-internal transcribed spacer 2 (PCR-RFLP-ITS2) and, for selected individuals, ITS2 sequences. A few specimens from Lupuna, Loreto Department, northern Amazonian Peru, were also identified as An. benarrochi B. A statistical parsimony network using ITS2 sequences confirmed that all Peruvian An. benarrochi B analyzed were identical to those in GenBank from Putumayo, southern Colombia. Sequences of the mtDNA COI BOLD region of specimens from all three Peruvian localities were connected using a statistical parsimony network, although there were multiple mutation steps between northern and southern Peruvian sequences. A Bayesian inference of concatenated Peruvian sequences of ITS2 + COI detected a single clade with very high support for all An. benarrochi B except one individual from Lupuna that was excluded. No samples were positive for Plasmodium by CytB-PCR.

  12. Mosquito repellent potential of Pithecellobium dulce leaf and seed against malaria vector Anopheles stephensi (Diptera: Culicidae

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    Mohan Rajeswary

    2016-03-01

    Full Text Available Objective: To determine the repellent properties of hexane, benzene, ethyl acetate, chloroform and methanol extract of Pithecellobium dulce (P. dulce leaf and seed against Anopheles stephensi (An. stephensi. Methods: Repellent activity assay was carried out in a net cage (45 cm × 30 cm × 25 cm containing 100 blood starved female mosquitoes of An. stephensi. This assay was carried out in the laboratory conditions according to the WHO 2009 protocol. Plant crude extracts of P. dulce were applied at 1.0, 2.5, and 5.0 mg/cm2 separately in the exposed fore arm of study subjects. Ethanol was used as the sole control. Results: In this study, the applied plant crude extracts were observed to protect against mosquito bites. There were no allergic reactions experienced by the study subjects. The repellent activity of the extract was dependent on the concentration of the extract. Among the tested solvents, the leaf and seed methanol extract showed the maximum efficacy. The highest concentration of 5.0 mg/cm2 leaf and seed methanol extract of P. dulce provided over 180 min and 150 min protection, respectively. Conclusions: Crude extracts of P. dulce exhibit the potential for controlling malaria vector mosquito An. stephensi.

  13. Isoenzymatic analysis of four Anopheles (Kerteszia) bellator Dyar & Knab (Diptera: Culicidae) populations.

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    de Carvalho-Pinto, Carlos José; Lourenço-de-Oliveira, Ricardo

    2003-12-01

    Anopheles bellator is a small silvatic bromelia-breeding mosquito and is a primary human malaria vector species in Southern Brazil. The bromelia-breeding habitat of the species should accompany the Atlantic forest coastal distribution, where bromeliads are abundant. Nonetheless, records on An. bellator collections show a gap in the species geographical distribution. An. bellator has been recorded in Southern Brazil and in the Brazilian states of Bahia and Paraíba. It appears again in the island of Trinidad, in Trinidad and Tobago. The aim of this work was to measure gene flow between different populations of An. bellator collected in the northern and southern extremes of the geographic distribution of this species. Mosquitoes were captured in forest borders in Santa Catarina, São Paulo, and Bahia states in Brazil and in the island of Trinidad in Republic of Trinidad and Tobago. Genetic distances varied between 0.076 and 0.680, based on enzymatic profiles from 11 distinct isoenzymes. Results indicate the existence of low-level gene flow between Brazilian populations of An. bellator, and a gene flow was even lower between the Brazilian and the Trinidad populations. This finding lead us to hypothesize that An. bellator did not spread along the coast, but reached northeastern areas through inland routes.

  14. Isoenzymatic analysis of four Anopheles (Kerteszia bellator Dyar & Knab (Diptera: Culicidae populations

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    Carvalho-Pinto Carlos José de

    2003-01-01

    Full Text Available Anopheles bellator is a small silvatic bromelia-breeding mosquito and is a primary human malaria vector species in Southern Brazil. The bromelia-breeding habitat of the species should accompany the Atlantic forest coastal distribution, where bromeliads are abundant. Nonetheless, records on An. bellator collections show a gap in the species geographical distribution. An. bellator has been recorded in Southern Brazil and in the Brazilian states of Bahia and Paraíba. It appers again in the island of Trinidad, in Trinidad and Tobago. The aim of this work was to measure gene flow between different populations of An. bellator collected in the northern and southern extremes of the geographic distribution of this species. Mosquitoes were captured in forest borders in Santa Catarina, São Paulo, and Bahia states in Brazil and in the island of Trinidad in Republic of Trinidad and Tobago. Genetic distances varied between 0.076 and 0.680, based on enzymatic profiles from 11 distinct isoenzymes. Results indicate the existence of low-level gene flow between Brazilian populations of An. bellator, and a gene flow was even lower between the Brazilian and the Trinidad populations. This finding lead us to hypothesize that An. bellator did not spread along the coast, but reached northeastern areas through inland routes.

  15. Larvicidal effects of endophytic and basidiomycete fungus extracts on Aedes and Anopheles larvae (Diptera, Culicidae

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    Augusto Bucker

    2013-08-01

    Full Text Available Introduction In vitro bioassays were performed to access the larvicidal activity of crude extracts from the endophytic fungus Pestalotiopsis virgulata (Melanconiales, Amphisphaeriaceae and the saprophytic fungus Pycnoporus sanguineus (Basidiomycetes, Polyporaceae against the mosquitoes Aedes aegypti and Anopheles nuneztovari. Methods The extracts were tested at concentrations of 100, 200, 300, 400 and 500ppm. Ethyl acetate mycelia (EAM extracts and liquid culture media (LCM from Pe. virgulata and Py. sanguineus were tested against third instar larvae of Ae. aegypti and An. nuneztovari. Results The larvicidal activity of the EAM extracts from Pe. virgulata against Ae. aegypti had an LC50=101.8ppm, and the extract from the basidiomycete fungus Py. sanguineus had an LC50=156.8ppm against the Ae. aegypti larvae. The Pe. virgulata extract had an LC50=16.3ppm against the An. nuneztovari larvae, and the Py. sanguineus extract had an LC50=87.2ppm against these larvae. Conclusions These results highlight the larvicidal effect of EAM extracts from the endophyte Pe. virgulata against the two larval mosquitoes tested. Thus, Pe. virgulata and Py. sanguineus have the potential for the production of bioactive substances against larvae of these two tropical disease vectors, with An. nuneztovari being more susceptible to these extracts.

  16. The fine-scale genetic structure of the malaria vectors Anopheles funestus and Anopheles gambiae (Diptera: Culicidae) in the north-eastern part of Tanzania

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    Gélin, P.; Magalon, H.; Drakeley, C.; Maxwell, C.; Magesa, S.; Takken, W.; Boëte, C.

    2016-01-01

    Understanding the impact of altitude and ecological heterogeneity at a fine scale on the populations of malaria vectors is essential to better understand and anticipate eventual epidemiological changes. It could help to evaluate the spread of alleles conferring resistance to insecticides and also

  17. Variability and genetic differentiation among Anopheles (Ano. intermedius Chagas, 1908 and Anopheles (Ano. mattogrossensis Lutz & Neiva, 1911 (Diptera: Culicidae from the Brazilian Amazon

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    Joselita Maria Mendes dos Santos

    2005-10-01

    Full Text Available Anopheles (Anopheles intermedius and Anopheles (Ano. mattogrossensis are Brazilian anopheline species belonging to the scarcely studied Anopheles subgenus. Few studies have been done on the genetic differentiation of these species. Both species have been found infected by Plasmodium and are sympatric with other anopheline species from the Nyssorhynchus subgenus. Eighteen enzymatic loci were analyzed in larval specimens of An. intermedius and An. mattogrossensis aiming to estimate the variability and genetic differentiation between these species. An. mattogrossensis population showed higher genetic variability (P = 44.4 and Ho = 0.081 ± 0.031 than that of An. intermedius (P = 33.3 and Ho = 0.048 ± 0.021. Most analyzed loci showed genotypic frequencies according to Hardy-Weinberg equilibrium, except for LAP1 and LAP2 in An. intermedius, and EST1 and PGM loci in An. mattogrossensis. The genetic distance between these species (D = 0.683 was consistent with the inter-specific values reported for Anopheles subgenus. We verified that the polymorphism and heterozygosity percentile values found in both species and compared to those in the literature, showed no relation between the level of isozyme variability and geographical distribution. The low variability found in these two species is probably more related to the niche they occupy than to their geographic distribution.

  18. Update on resistance status of Anopheles gambiae s.s. to conventional insecticides at a previous WHOPES field site, "Yaokoffikro", 6 years after the political crisis in Côte d'Ivoire

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    Koffi Alphonsine A

    2012-04-01

    Full Text Available Abstract Background At Yaokoffikro field site near Bouaké, in central Côte d'Ivoire, a group of experimental huts built in 1996 served over many years for the evaluation of insecticides against highly resistant mosquitoes. Breeding sites of mosquitoes and selection pressure in the area were maintained by local farming practices until a war broke out in September 2002. Six years after the crisis, we conducted bioassays and biochemical analysis to update the resistance status of Anopheles gambiae s.s. populations and detect other potential mechanisms of resistance that might have evolved. Methods An. gambiae s.s. larvae from Yaokoffikro were collected in breeding sites and reared to adults. Resistance status of this population to insecticides was assessed using WHO bioassay test kits for adult mosquitoes with seven insecticides: two pyrethroids, a pseudo-pyrethroid, an organochloride, two carbamates and an organophosphate. Molecular and biochemical assays were carried out to identify the L1014F kdr and ace-1R alleles in individual mosquitoes and to detect potential increase in mixed function oxidases (MFO, non-specific esterases (NSE and glutathione S-transferases (GST activity. Results High pyrethroids, DDT and carbamate resistance was confirmed in An. gambiae s.s. populations from Yaokoffikro. Mortality rates were less than 70% with pyrethroids and etofenprox, 12% with DDT, and less than 22% with the carbamates. Tolerance to fenitrothion was observed, with 95% mortality after 24 h. PCR analysis of samples from the site showed high allelic frequency of the L1014F kdr (0.94 and the ace-1R (0.50 as before the crisis. In addition, increased activity of NSE, GST and to a lesser extent MFO was found relative to the reference strain Kisumu. This was the first report detecting enhanced activity of these enzymes in An. gambiae s.s from Yaokoffikro, which could have serious implications in detoxification of insecticides. Their specific roles in

  19. Control of pyrethroid and DDT-resistant Anopheles gambiae by application of indoor residual spraying or mosquito nets treated with a long-lasting organophosphate insecticide, chlorpyrifos-methyl

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    Chabi Joseph

    2010-02-01

    Full Text Available Abstract Background Scaling up of long-lasting insecticidal nets (LLINs and indoor residual spraying (IRS with support from the Global Fund and President's Malaria Initiative is providing increased opportunities for malaria control in Africa. The most cost-effective and longest-lasting residual insecticide DDT is also the most environmentally persistent. Alternative residual insecticides exist, but are too short-lived or too expensive to sustain. Dow Agrosciences have developed a microencapsulated formulation (CS of the organophosphate chlorpyrifos methyl as a cost-effective, long-lasting alternative to DDT. Methods Chlorpyrifos methyl CS was tested as an IRS or ITN treatment in experimental huts in an area of Benin where Anopheles gambiae and Culex quinquefasiactus are resistant to pyrethroids, but susceptible to organophosphates. Efficacy and residual activity was compared to that of DDT and the pyrethroid lambdacyalothrin. Results IRS with chlorpyrifos methyl killed 95% of An. gambiae that entered the hut as compared to 31% with lambdacyhalothrin and 50% with DDT. Control of Cx. quinquefasciatus showed a similar trend; although the level of mortality with chlorpyrifos methyl was lower (66% it was still much higher than for DDT (14% or pyrethroid (15% treatments. Nets impregnated with lambdacyhalothrin were compromized by resistance, killing only 30% of An. gambiae and 8% of Cx. quinquefasciatus. Nets impregnated with chlorpyrifos methyl killed more (45% of An gambiae and 15% of Cx. quinquefasciatus, but its activity on netting was of short duration. Contact bioassays on the sprayed cement-sand walls over the nine months of monitoring showed no loss of activity of chlorpyrifos methyl, whereas lambdacyhalothrin and DDT lost activity within a few months of spraying. Conclusion As an IRS treatment against pyrethroid resistant mosquitoes chlorpyrifos methyl CS outperformed DDT and lambdacyhalothrin. In IRS campaigns, chlorpyrifos methyl CS should

  20. Chemical Constituents and Combined Larvicidal Effects of Selected Essential Oils against Anopheles cracens (Diptera: Culicidae

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    Jitrawadee Intirach

    2012-01-01

    Full Text Available A preliminary study on larvicidal activity against laboratory-colonized Anopheles cracens mosquitos revealed that five of ten plant oils at concentration of 100 ppm showed 95–100% larval mortality. The essential oils of five plants, including Piper sarmentosum, Foeniculum vulgare, Curcuma longa, Myristica fragrans, and Zanthoxylum piperitum, were then selected for chemical analysis, dose-response larvicidal experiments, and combination-based bioassays. Chemical compositions analyzed by gas chromatography coupled to mass spectrometry demonstrated that the main component in the oil derived from P. sarmentosum, F. vulgare, C. longa, M. fragrans, and Z. piperitum was croweacin (71.01%, anethole (63.00%, ar-turmerone (30.19%, safrole (46.60%, and 1,8-cineole (21.27%, respectively. For larvicidal bioassay, all five essential oils exerted promising efficacy in a dose-dependent manner and different performances on A. cracens after 24 hours of exposure. The strongest larvicidal potential was established from P. sarmentosum, followed by F. vulgare, C. longa, M. fragrans, and Z. piperitum, with LC50 values of 16.03, 32.77, 33.61, 40.00, and 63.17 ppm, respectively. Binary mixtures between P. sarmentosum, the most effective oil, and the others at the highest ratio were proved to be highly efficacious with a cotoxicity coefficient value greater than 100, indicating synergistic activity. Results of mixed formulations of different essential oils generating synergistic effects may prove helpful in developing effective, economical, and ecofriendly larvicides, as favorable alternatives for mosquito management.

  1. Larvicidal activity of indigenous plant extracts on the rural malarial vector, Anopheles culicifacies Giles. (Diptera: Culicidae

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    K. Kovendan

    2014-12-01

    Full Text Available Vector control is one of the most important components in combating vector-borne diseases throughout the world. Application of insecticides is a widely known and popular vector control strategy. The objective of the present study was to evaluate the larvicidal activity of the hexane, diethyl ether, ethyl acetate and acetone extracts of Abutilon indicum, Hyptis suaveolens and Leucas aspera against third-stage larvae of Anopheles culicifiacies. The results clearly suggest that all three selected plant extracts exhibited moderate larvicidal activity after 24, 48 and 72 h at 250, 500, 750 and 1000 ppm; the lethal concentrations (LC at 50% and 90% of A. indicum, H. suaveolens against third instar larvae at 24, 48 and 72 h (hexane, diethyl ether, ethyl acetate and acetone were as follows: A. indicum, LC50=1031.65, 949.18, 833.58 and 673.68 ppm; LC90=2215.87, 2234.39, 2152.97 and 2455.10 ppm; H. suaveolens, LC50=423.00, 347.50, 236.58 and 217.24 ppm; LC90=1431.91, 1292.15, 1138.49 and 1049.27 ppm and L. aspera, LC50=559.77, 401.56, 299.71 and 263.01 ppm; LC90=1400.80, 1549.31, 1157.96 and 1108.72 ppm at 24 h, respectively. Overall, the highest larvicidal activity was observed with H. suaveolens extract followed by L. aspera and A. indicum at various concentrations at 48 and 72 h, respectively. The objective of this investigation was an attempt to search for a user- and eco-friendly vector control agent. The study proved that the selected plant leaf extracts could serve as potent larvicidal agents against A. culicifacies in vector control programs.

  2. Antimalarial efficacy of dynamic compound of plumbagin chemical constituent from Plumbago zeylanica Linn (Plumbaginaceae) against the malarial vector Anopheles stephensi Liston (Diptera: Culicidae).

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    Pradeepa, Venkatraman; Sathish-Narayanan, Subbiah; Kirubakaran, Suyambulingam Arunachalam; Senthil-Nathan, Sengottayan

    2014-08-01

    In the present investigation, the effective root compound of plumbagin of Plumbago zeylanica (Plumbaginaceae) was evaluated for chemical constituent and antimalarial effect against the fourth instar larvae of Anopheles stephensi Liston (Diptera). In the chromatographic analyses of root compound with Rf value of 0.788 and NMR analyses also revealed that the effective compound contain naphthoquinone plumbagin were identified as the major chemical constituent. Larval mortality was observed after 3 h of exposure period. The plumbagin compound showed remarkable larvicidal activity against A. stephensi (LC50 32.65 and LC9072.27 ppm). Histopathological effects of compound was observed in the treated larvae. Based on the results, the plumbagin compound of P. zeylanica can be considered as a new source of natural larvicide for the control of malarial vector.

  3. Lineage divergence detected in the malaria vector Anopheles marajoara (Diptera: Culicidae in Amazonian Brazil

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    Povoa Marinete M

    2010-10-01

    Full Text Available Abstract Background Cryptic species complexes are common among anophelines. Previous phylogenetic analysis based on the complete mtDNA COI gene sequences detected paraphyly in the Neotropical malaria vector Anopheles marajoara. The "Folmer region" detects a single taxon using a 3% divergence threshold. Methods To test the paraphyletic hypothesis and examine the utility of the Folmer region, genealogical trees based on a concatenated (white + 3' COI sequences dataset and pairwise differentiation of COI fragments were examined. The population structure and demographic history were based on partial COI sequences for 294 individuals from 14 localities in Amazonian Brazil. 109 individuals from 12 localities were sequenced for the nDNA white gene, and 57 individuals from 11 localities were sequenced for the ribosomal DNA (rDNA internal transcribed spacer 2 (ITS2. Results Distinct A. marajoara lineages were detected by combined genealogical analysis and were also supported among COI haplotypes using a median joining network and AMOVA, with time since divergence during the Pleistocene (COI sequences at the 3' end were more variable, demonstrating significant pairwise differentiation (3.82% compared to the more moderate 2.92% detected by the Folmer region. Lineage 1 was present in all localities, whereas lineage 2 was restricted mainly to the west. Mismatch distributions for both lineages were bimodal, likely due to multiple colonization events and spatial expansion (~798 - 81,045 ya. There appears to be gene flow within, not between lineages, and a partial barrier was detected near Rio Jari in Amapá state, separating western and eastern populations. In contrast, both nDNA data sets (white gene sequences with or without the retention of the 4th intron, and ITS2 sequences and length detected a single A. marajoara lineage. Conclusions Strong support for combined data with significant differentiation detected in the COI and absent in the nDNA suggest that

  4. Confirmation of Anopheles (Anopheles calderoni Wilkerson, 1991 (Diptera: Culicidae in Colombia and Ecuador through molecular and morphological correlation with topotypic material

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    Ranulfo González

    2010-12-01

    Full Text Available The morphologically similar taxa Anopheles calderoni, Anopheles punctimacula, Anopheles malefactor and Anopheles guarao are commonly misidentified. Isofamilies collected in Valle de Cauca, Colombia, showed morphological characters most similar to An. calderoni, a species which has never previously been reported in Colombia. Although discontinuity of the postsubcostal pale spots on the costa (C and first radial (R1 wing veins is purportedly diagnostic for An. calderoni, the degree of overlap of the distal postsubcostal spot on C and R1 were variable in Colombian specimens (0.003-0.024. In addition, in 98.2% of larvae, seta 1-X was located off the saddle and seta 3-C had 4-7 branches in 86.7% of specimens examined. Correlation of DNA sequences of the second internal transcribed spacer and mtDNA cytochrome c oxidase subunit I gene (COI barcodes (658 bp of the COI gene generated from Colombian progeny material and wild-caught mosquitoes from Ecuador with those from the Peruvian type series of An. calderoni confirmed new country records. DNA barcodes generated for the closely related taxa, An. malefactor and An. punctimacula are also presented for the first time. Examination of museum specimens at the University of the Valle, Colombia, revealed the presence of An. calderoni in inland localities across Colombia and at elevations up to 1113 m.

  5. Distribution and frequency of kdr mutations within Anopheles gambiae s.l. populations and first report of the ace.1 G119S mutation in Anopheles arabiensis from Burkina Faso (West Africa.

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    Roch K Dabiré

    Full Text Available An entomological survey was carried out at 15 sites dispersed throughout the three eco-climatic regions of Burkina Faso (West Africa in order to assess the current distribution and frequency of mutations that confer resistance to insecticides in An. gambiae s.l. populations in the country. Both knockdown (kdr resistance mutation variants (L1014F and L1014S, that confer resistance to pyrethroid insecticides, were identified concomitant with the ace-1 G119S mutation confirming the presence of multiple resistance mechanisms in the An. gambiae complex in Burkina Faso. Compared to the last survey, the frequency of the L1014F kdr mutation appears to have remained largely stable and relatively high in all species. In contrast, the distribution and frequency of the L1014S mutation has increased significantly in An. gambiae s.l. across much of the country. Furthermore we report, for the first time, the identification of the ace.1 G119S mutation in An. arabiensis populations collected at 8 sites [corrected]. This mutation, which confers resistance to organophosphate and carbamate insecticides, has been reported previously only in the An. gambiae S and M molecular forms. This finding is significant as organophosphates and carbamates are used in indoor residual sprays (IRS to control malaria vectors as complementary strategies to the use of pyrethroid impregnated bednets. The occurrence of the three target-site resistance mutations in both An. gambiae molecular forms and now An. arabiensis has significant implications for the control of malaria vector populations in Burkina Faso and for resistance management strategies based on the rotation of insecticides with different modes of action.

  6. Quantification of the efficiency of treatment of Anopheles gambiae breeding sites with petroleum products by local communities in areas of insecticide resistance in the Republic of Benin

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    Doannio Julien MC MC

    2007-05-01

    Full Text Available Abstract Background The emergence of Anopheles populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for developing new insecticidal molecules or, improving the efficacy of existing insecticides or abandoning those to which resistance has emerged. The use of petroleum products (PP against mosquito larvae had an immense success during early programmes of malaria control, but these compounds were abandoned and replaced in the 1950s by synthetic insecticides probably because of the high performances given by these new products. In the current context of vector resistance, it is important to elucidate the empirical use of PP by quantifying their efficiencies on resistant strains of Anopheles. Methods Larvae of Anopheles Ladji a local resistant strain were exposed to increasing concentrations of various PP (kerosene, petrol and engine oils for 24 hours and the lethal activities recorded. The highest concentration (HiC having no lethal activity (also referred as the NOEL or no effect level and the lowest concentration (LoC100 yielding 100% mortality were rated for each PP on the Ladji strain. Prior to laboratory analysis, KAP studies were conducted in three traditional communities were insecticide resistance is clearly established to confirm the use of PP against mosquitoes. Results Laboratory analysis of petrol, kerosene and engine oils, clearly established their lethal activities on resistant strains of Anopheles larvae. Contrary to existing references, this research revealed that exposed larvae of Anopheles were mostly killed by direct contact toxicity and not by suffocation as indicated in some earlier reports. Conclusion This research could serve as scientific basis to backup the empirical utilisation of PP on mosquito larvae and to envisage possibilities of using PP in some traditional settings where Anopheles have developed

  7. Experimental hut evaluation of bednets treated with an organophosphate (chlorpyrifos-methyl or a pyrethroid (lambdacyhalothrin alone and in combination against insecticide-resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes

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    Corbel Vincent

    2005-05-01

    Full Text Available Abstract Background Pyrethroid resistant mosquitoes are becoming increasingly common in parts of Africa. It is important to identify alternative insecticides which, if necessary, could be used to replace or supplement the pyrethroids for use on treated nets. Certain compounds of an earlier generation of insecticides, the organophosphates may have potential as net treatments. Methods Comparative studies of chlorpyrifos-methyl (CM, an organophosphate with low mammalian toxicity, and lambdacyhalothrin (L, a pyrethroid, were conducted in experimental huts in Côte d'Ivoire, West Africa. Anopheles gambiae and Culex quinquefasciatus mosquitoes from the area are resistant to pyrethroids and organophosphates (kdr and insensitive acetylcholinesterase Ace.1R. Several treatments and application rates on intact or holed nets were evaluated, including single treatments, mixtures, and differential wall/ceiling treatments. Results and Conclusion All of the treatments were effective in reducing blood feeding from sleepers under the nets and in killing both species of mosquito, despite the presence of the kdr and Ace.1R genes at high frequency. In most cases, the effects of the various treatments did not differ significantly. Five washes of the nets in soap solution did not reduce the impact of the insecticides on A. gambiae mortality, but did lead to an increase in blood feeding. The three combinations performed no differently from the single insecticide treatments, but the low dose mixture performed encouragingly well indicating that such combinations might be used for controlling insecticide resistant mosquitoes. Mortality of mosquitoes that carried both Ace.1R and Ace.1S genes did not differ significantly from mosquitoes that carried only Ace.1S genes on any of the treated nets, indicating that the Ace.1R allele does not confer effective resistance to chlorpyrifos-methyl under the realistic conditions of an experimental hut.

  8. IgG1 and IgG4 antibody responses to the Anopheles gambiae salivary protein gSG6 in the sympatric ethnic groups Mossi and Fulani in a malaria hyperhendemic area of Burkina Faso.

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    Cinzia Rizzo

    Full Text Available Human antibody response to the Anopheles gambiae salivary protein gSG6 has recently emerged as a potentially useful tool for malaria epidemiological studies and for the evaluation of vector control interventions. However, the current understanding of the host immune response to mosquito salivary proteins and of the possible crosstalk with early response to Plasmodium parasites is still very limited. We report here the analysis of IgG1 and IgG4 subclasses among anti-gSG6 IgG responders belonging to Mossi and Fulani from Burkina Faso, two ethnic groups which are known for their differential humoral response to parasite antigens and for their different susceptibility to malaria. The IgG1 antibody response against the gSG6 protein was comparable in the two groups. On the contrary, IgG4 titers were significantly higher in the Fulani where, in addition, anti-gSG6 IgG4 antibodies appeared in younger children and the ratio IgG4/IgG1 stayed relatively stable throughout adulthood. Both gSG6-specific IgG1 and IgG4 antibodies showed a tendency to decrease with age whereas, as expected, the IgG response to the Plasmodium circumsporozoite protein (CSP exhibited an opposite trend in the same individuals. These observations are in line with the idea that the An. gambiae gSG6 salivary protein induces immune tolerance, especially after intense and prolonged exposure as is the case for the area under study, suggesting that gSG6 may trigger in exposed individuals a Th2-oriented immune response.

  9. Fauna and some biological characteristics of Anopheles mosquitoes (Diptera:Culicidae) in Kalaleh County, Golestan Province, northeast of lran

    Institute of Scientific and Technical Information of China (English)

    Aioub Sofizadeh; Hamideh Edalat; Mohammad Reza Abai; Ahmad Ali Hanafi-Bojd

    2016-01-01

    Objective: To determine fauna and some ecological aspects of Anopheles mosquitoes in northeast of Iran. Methods: In this descriptive study, 3 villages in Kalaleh County were selected in different geographical zones. Anopheles mosquitoes were collected biweekly from May to October using standard dipping method for larvae, and hand catch, total catch, artificial pit shelter as well as night-biting collections on human and animal baits for adults. Results: Totally 399 larvae and 2 602 adults of Anopheles mosquitoes were collected and identified as 2 species: Anopheles superpictus s.l. (An. superpictus s.l.) and Anopheles maculipennis s.l. The dominant species was An. superpictus s.l. (92.1%). Activity of these mosquitoes found to be started from middle of May and extended till September with two peaks of activity in July and August. Conclusions: An. superpictus s.l. as one of the main malaria vectors in Iran as well as some other parts of the world is the dominant species in the study area. This species has high potential for transmission and possibility of establishing a transmission cycle with low abundance. Other species, Anopheles maculipennis s.l. also has introduced as a malaria vector in northern parts of Iran. As this Anopheles is a complex species, genetic studies are recommended to determine the members of this complex in the study area.

  10. Change in composition of the Anopheles gambiae complex and its possible implications for the transmission of malaria and lymphatic filariasis in north-eastern Tanzania

    DEFF Research Database (Denmark)

    Derua, Yahya A; Alifrangis, Michael; Hosea, Kenneth M;

    2012-01-01

    ABSTRACT: BACKGROUND: A dramatic decline in the incidence of malaria due to Plasmodium falciparum infection in coastal East Africa has recently been reported to be paralleled (or even preceded) by an equally dramatic decline in malaria vector density, despite absence of organized vector control. ...... to differences in biology and vectorial capacity of the An. gambiae s.l. complex the change in sibling species composition will have important implications for the epidemiology and control of malaria and lymphatic filariasis in the study area....

  11. Ovicidal, larvicidal and adulticidal properties of Asparagus racemosus (Willd.) (Family: Asparagaceae) root extracts against filariasis (Culex quinquefasciatus), dengue (Aedes aegypti) and malaria (Anopheles stephensi) vector mosquitoes (Diptera: Culicidae).

    Science.gov (United States)

    Govindarajan, Marimuthu; Sivakumar, Rajamohan

    2014-04-01

    Several diseases are associated to the mosquito-human interaction. Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikungunya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present investigation was undertaken to study the ovicidal, larvicidal and adulticidal activities of crude hexane, ethyl acetate, benzene, chloroform and methanol extracts of root of Asparagus racemosus were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae). The mean percent hatchability of the eggs was observed after 48 h post-treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the methanol extract showed the highest ovicidal activity. The methanol extract of Asparagus racemosus against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi exerted 100% mortality (zero hatchability) at 375, 300 and 225 ppm, respectively. Control eggs showed 99-100% hatchability. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in methanol extract of root of Asparagus racemosus against the larvae of Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi with the LC50 and LC90 values were 115.13, 97.71 and 90.97 ppm and 210.96, 179.92, and 168.82 ppm, respectively. The adult mortality was observed after 24 h recovery period. The plant crude extracts showed dose-dependent mortality. At higher concentrations, the adult showed restless movement for some times with abnormal wagging and then died. Among the extracts tested, the highest adulticidal activity was observed in

  12. Efficacy of sunlight-activatable porphyrin formulates on larvae of Anopheles gambiae M and S molecular forms and An. arabiensis: a potential novel biolarvicide for integrated malaria vector control.

    Science.gov (United States)

    Fabris, Clara; Ouédraogo, Robert Kossivi; Coppellotti, Olimpia; Dabiré, Roch K; Diabaté, Abdoulaye; Di Martino, Piera; Guidolin, Laura; Jori, Giulio; Lucantoni, Leonardo; Lupidi, Giulio; Martena, Valentina; Sawadogo, Simon P; Soncin, Marina; Habluetzel, Annette

    2012-09-01

    Biolarvicides, such as microbial formulations based on Bacillus thuringiensis and B. sphaericus, have been found to be highly effective against mosquito larvae and are currently employed as eco-friendly alternatives to synthetic chemical insecticides for vector control. Recently, a porphyrin of natural origin has been suggested as a sunlight-activatable larvicide against the dengue vector Aedes aegypti. In order to validate the approach for the control of the malaria vector, we tested the photo-larvicidal activity of a novel porphyrin, namely meso-tri(N-methyl-pyridyl), mono(N-dodecyl-pyridyl)porphine, C12, associated with two specifically selected carriers, against Anopheles gambiae s.s. and An. arabiensis larvae, both laboratory reared and collected from malaria endemic sites in Burkina Faso. Both C12-porphyrin formulates, when administered to larvae at a 50μM porphyrin dose, were accumulated in the alimentary canal. Subsequent exposure of the porphyrin-loaded larvae to sunlight for short times (0.5-3h) led to a complete mortality. The high efficacy exhibited by a "foodstuff" porphyrin formulate also in the presence of typical larval food particles opens promising perspectives for the development of an effective photocidal larvicide.

  13. Development of an allele-specific, loop-mediated, isothermal amplification method (AS-LAMP to detect the L1014F kdr-w mutation in Anopheles gambiae s. l.

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    Badolo Athanase

    2012-07-01

    Full Text Available Abstract Background Malaria control relies heavily on treated bed nets and indoor residual spraying with pyrethroid insecticides. Unfortunately, the resistance to pyrethroid insecticides, mainly due to the kdr mutation, is spreading in the main malaria vector Anopheles gambiae s.l., decreasing the insecticides’ efficacy. To manage the insecticide resistance rapidly and flexibly, simple and effective tools for the early detection of resistant mosquitoes are needed. This study aimed to develop an allele-specific, loop-mediated, isothermal amplification (AS-LAMP method to detect the West African-type kdr mutation (kdr-w; L1014F in field-collected mosquitoes. Methods DNA fragments of the wild-type and the mutated kdr gene were used to select the primers and develop the method. The primers were designed with the mutation at the 5’ end of the backward inner primer (BIP. The AS-LAMP method was compared to the AS-PCR method using the genomic DNA of 120 field-collected mosquitoes. Results The AS-LAMP method could discriminate between the wild-type homozygote, the heterozygote, and the kdr-w homozygote within 75 min. The AS-LAMP method has the advantage of being faster and at least as sensitive and specific as the AS-PCR method. Conclusions The AS-LAMP method can be used to detect the kdr mutation for quick decision-making, even in less well-equipped laboratories.

  14. Differential effect of human ivermectin treatment on blood feeding Anopheles gambiae and Culex quinquefasciatus

    DEFF Research Database (Denmark)

    Derua, Yahya A.; Kisinza, William N.; Simonsen, Paul Erik

    2015-01-01

    BACKGROUND: Widespread and large scale use of ivermectin in humans and domestic animals can have unexpected effects on non-target organisms. As a search for a possible explanation for an observed longitudinal decline in density of anopheline vector mosquitoes, but not in Culex quinquefasciatus...... to receive either ivermectin or placebo. Twenty four hours after treatment, one volunteer from each group was concurrently exposed to 50 laboratory reared An. gambiae on one arm and 50 laboratory reared Cx. quinquefasciatus on the other arm for 15-30 minutes. Engorged mosquitoes were maintained on 10...

  15. IgG responses to Anopheles gambiae salivary antigen gSG6 detect variation in exposure to malaria vectors and disease risk

    DEFF Research Database (Denmark)

    Stone, Will; Bousema, Teun; Jones, Sophie;

    2012-01-01

    , IgG responses to gSG6 in individual children showed a strong positive association with household level mosquito exposure. IgG levels for all antigens except AMA-1 were associated with the frequency of malaria episodes following sampling. gSG6 seropositivity was strongly positively associated...... with subsequent malaria incidence (test for trend p¿=¿0.004), comparable to malaria antigens MSP-1 and GLURP R2. Our results show that the gSG6 assay is sensitive to micro-epidemiological variations in exposure to Anopheles mosquitoes, and provides a correlate of malaria risk that is unrelated to immune...

  16. Electrophysiological responses of gustatory receptor neurons on the labella of the common malaria mosquito Anopheles quadrimaculatus Say (Diptera: Culicidae)

    Science.gov (United States)

    We recorded electrical responses from sensory cells associated with gustatory sensilla on the labella of female Anopheles quadrimaculatus to salt, sucrose, quinine (a feeding deterrent) and the insect repellent, N,N-diethyl-3-methylbenzamide (DEET). A salt-sensitive cell responded to increasing con...

  17. Effect of Bacillus sphaericus Neide on Anopheles (Diptera: Culicidae and associated insect fauna in fish ponds in the Amazon

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    Francisco Augusto da Silva Ferreira

    2015-09-01

    Full Text Available ABSTRACTWe analyzed the effects of Bacillus sphaericus on Anopheles larvae and on the associated insect fauna in fish farming ponds. Five breeding sites in the peri-urban area of the city of Manaus, AM, Brazil, were studied. Seven samples were collected from each breeding site and B. sphaericus was applied and reapplied after 15 days. The samples were made at 24 h before application, 24 h post-application and 5 and 15 days post-application. We determined abundance, larval reduction and larval density for Anopheles, and abundance, richness, Shannon diversity index and classified according to the functional trophic groups for associated insect fauna. A total of 904 Anopheles larvae were collected and distributed into five species. Density data and larval reduction demonstrated the rapid effect of the biolarvicide 24 h after application. A total of 4874 associated aquatic insects belonging to six orders and 23 families were collected. Regression analysis of diversity and richness indicated that the application of the biolarvicide had no influence on these indices and thus no effect on the associated insect fauna for a period of 30 days. B. sphaericus was found to be highly effective against the larvae of Anopheles, eliminating the larvae in the first days after application, with no effect on the associated insect fauna present in the fish ponds analyzed.

  18. Man-biting activity of Anopheles (Nyssorhynchus albimanus and An. (Kerteszia neivai (Diptera: Culicidae in the Pacific Lowlands of Colombia

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    Yezid Solarte

    1996-04-01

    Full Text Available The daily man-biting activity of Anopheles (Nyssorhynchus albimanus and An. (Kerteszia neivai was determined in four ecologically distinct settlements of the Naya River, Department of Valle, Colombia. Differences were found among the settlements with respect to the mosquito species present, intradomiciliary and extradomiciliary biting activity and population densities.

  19. Mosquitocidal activity of Polygala arvensis Willd against Aedes aegypti (Linn., Anopheles stephensi (Liston. and Culex quinquefasciatus (Say. (Diptera: Culicidae

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    M. Deepa

    2014-12-01

    Full Text Available To determine the larvicidal, ovicidal and repellent activities of benzene and methanol extract of Polygala arvensis against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus Twenty five 3rd instar larvae of selected mosquitoes species were exposed to various concentrations (60-300 ppm and were assayed in the laboratory by using the protocol of WHO 2005; the 24 h LC50 values of the P. Arvensis leaf extract was determined following Probit analysis. The ovicidal activity was determined against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus exposed to variousconcentrations were tested under laboratory conditions and the hatch rates were assessed 120hrs post treatment. The repellent efficacy was determined against selected mosquitoes at three concentrations viz., 1.0, 2.0 and 3.0 mg/cm2 under the laboratory conditions. The LC50 and LC90 values of benzene and methanol extract of P. arvensis against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus larvae in 24 h were 75.32, 88.26, 82.46, 58.21, 46.37, 42.68 and 260.48, 275.26, 251.39, 208.45, 189.82 and 130.44 ppm, respectively. It has been noticed that the higher concentrations of P. arvensis extractspossesses strong ovicidal activity at 200 ppm concentration against Ae. aegypti, An. stephensi and C. quinquefasciatus, no egg hatchability was recorded. In the same way, methanol extracts showed maximum ovicidal activity followed by benzene extract against selected vector mosquitoes. In repellent activity, among two extracts tested P. arvensis methanol extract had strong repellent action against selected mosquitoes as it provided 100% protection against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus for 280min. From the results it can be concluded the P. arvensis extract was an excellent potential for controlling Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus mosquitoes.

  20. Mosquito larvicidal and ovicidal properties of Pemphis acidula Frost. (Lythraceae) against Culex tritaeniorhynchus Giles and Anopheles subpictus Grassi (Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    K. Samidurai

    2012-01-01

    Objective: This study was undertaken to assess the larvicidal and ovicidal potential of the crude methanol, benzene and acetone solvent extracts from the medicinal plant Pemphis acidula (Pe. acidula) against the medically important mosquito vectors, Culex tritaeniorhynchus (Cx. tritaeniorhynchus) and Anopheles subpictus (An. subpictus) were exposed to various concentrations and (Diptera: Culicidae). Methods: Twenty five late third instar of Cx. tritaeniorhynchus and An. subpictus were exposed to various concentrations and were assayed in the laboratory by using the protocol of WHO 2005. The larval mortality was observed 24 h of treatment. Hundred eggs of Cx. tritaeniorhynchus and An. subpictus were exposed to various concentrations and were assayed in the laboratory by using the protocol of Su and Mulla 1998. The ovicidal activity was observed 48 h of treatment. Results: The LC50 and LC90 values being 10.81and 20.64 and 22.10 and 43.71 ppm and hundred percent of egg mortality was observed at 350 and 400 ppm methanol extract of Pe. acidula against Cx. tritaeniorhynchus and An. subpictus, respectively. Conclusion: These results suggest that the leaf extracts have the potential to be used as an ideal ecofriendly approach for the control of mosquitoes.

  1. The Gambia.

    Science.gov (United States)

    1982-11-01

    Attention in this discussion of the Republic of the Gambia is directed to the following: the people; geography; history; government and political conditions; economy; foreign relations; and relations between the US and Gambia. In 1980 Gambia's population was estimated to be 619,052; the 1980-81 annual growth rate estimate was 2.8%. The infant mortality rate is 217/1000; life expectancy is 32 years for men and 34 years for women. Almost 85% of the population live in rural areas. The principal ethnic groups are Mandinka, Fula, Wolof, Jola, and Serahuli, English is the official language, although most Gambians speak their native language. The Gambia, located on the bulge of West Africa, is low lying, with a maximum altitude of 73 meters above sea level. From what is known of its early history, the Gambia was once part of the Empire of Ghana and the Kingdom of the Songhais. By the 16th century, Portuguese slave traders and gold seekers had settled in the lower river area. During the late 17th and throughout the 18th century, England and France struggled continously for political and commercial supremacy in the regions of the Senegal and Gambia rivers. The Gambia achieved independence on February 18, 1965, as a constitutional monarchy within the British Commonwealth. Shortly thereafter the government proposed conversion from a monarchy to a republic with an elected president replacing the British monarch as chief of state. In 1970 a referendum was approved by the required majority, and the Gambia became a republic on April 24th. The government is divided into independent executive, legislative, and judicial branches. Executive power is vested in a president, popularly elected for a 5 year term. The Gambia is 1 of the few African countries with a genuine multiparty system. There is no army. The economy is almost totally agricultural. The small manfacturing sector accounts for about 5% of the gross domestic product. Few foreign investors have taken advantage of the

  2. Morphological Analysis of Anopheles vagus Donitz, 1902 (Diptera : Culicidae in fresh water and brackish water habitats = Variasi Morfologi Anopheles vagus Donitz, 1902 (Diptera : Culicidae dari Habitat Air Tawar dan Air Payau

    Directory of Open Access Journals (Sweden)

    Siti Alfiah

    2014-10-01

    Full Text Available ENGLISHAbstractAnopheles subpictus had habitat variation and showed genetic difference. So, the variation of habitat of An. vagus may support the hypothesa that An. vagus had genetic and morphology variation, same as An. subpictus.The aimed of this research was analyze morphology and chaetotaxy difference between An. vagus in fresh water and brackish water. The subject of the study was An. vagus collected from Kesongo Village, Tuntang Subdistrict, Semarang (fresh water and Jatimalang Village, Purwodadi Subdistrict, Purworejo (brackish water. Anopheles vagus were collected and individually reared. One sample in every batch was used to make larvae skin, pupae skin and adult specimen of An. vagus. The result showed that there were intra and inter population variation between An. vagus in fresh water and brackish water. The variations were on the size and number of hair branches and filaments. The conclution of this research were the morphology and chaetotaxy of female An. vagus in fresh water and brackish water showed no different. Intra and interpopulation An. vagus in fresh water and brackish water were caused by the difference of geography location (allopatric speciation.INDONESIANVariasi habitat terjadi pada An. subpictus, variasi habitat yang berbeda menunjukkan variasi genetik yang berbeda. Oleh karena itu variasi habitat An. vagus diduga akan bepengaruh terhadap variasi genetik dan morfologi. Tujuan penelitian adalah menganalisis perbedaan morfologi dan kaetotaksi Anopheles vagus habitat air tawar dan air payau. Subyek penelitian adalah An. vagus habitat air tawar di Desa Kesongo, Kecamatan Tuntang, Kabupaten Semarang dan An. vagus habitat air payau di Desa Jatimalang, Kecamatan Purwodadi, Kabupaten Purworejo. Anopheles vagus yang diperoleh, di rearing secara individual. Tiap indukan diambil satu sampel keturunannya dan dibuat preparat skin larva, skin pupa dan nyamuk dewasa betina. Hasil menunjukkan bahwa Anopheles vagus betina habitat air

  3. Larvicidal and repellent properties of Adansonia digitata against medically important human malarial vector mosquito Anopheles stephensi (Diptera: Culicidae)

    OpenAIRE

    K. Krishnappa , K. Elumalai , S. Dhanasekaran & J. Gokulakrishnan

    2012-01-01

    Background & objectives: Development of plant-based alternative compounds for mosquito control has gainedimportance now-a-days, in view of increasing resistance in mosquito vectors to existing insecticides. The larvicidaland repellent activities of benzene, chloroform, hexane and methanol leaf extracts of Indian medicinal plant,Adansonia digitata were investigated against malarial vector, Anopheles stephensi.Methods: In all, 25 III instar larvae of An. stephensi were exposed to various concen...

  4. Baseline Susceptibility of Different Geographical Strains of Anopheles stephensi (Diptera: Culicidae) to Temephos in Malarious Areas of Iran

    OpenAIRE

    Aboozar Soltani; Hassan Vatandoost; Mohammad Ali Oshaghi; Ahmad Ali Enayati; Ahmad Raeisi; Mohammad Reza Eshraghian; Mohammad Mehdi Soltan-Dallal; Ahmad Ali Hanafi-Bojd; Mohammad Reza Abai; Fatemeh Rafi

    2013-01-01

    Background: Malaria still remains a public health problem in Iran. There are different vector control interventions such as insecticide spraying. The present study was carried out to determine the susceptibility status of Anopheles stephensi larvae to temephos as a national plan for monitoring and mapping of insecticide resistance Methods: Eight different localities in two main malarious provinces were determined as field collecting sites. Mosquitoes were collected from the field and reared i...

  5. Remotely-sensed land use patterns and the presence of Anopheles larvae (Diptera: Culicidae) in Sukabumi, West Java, Indonesia.

    Science.gov (United States)

    Stoops, Craig A; Gionar, Yoyo R; Shinta; Sismadi, Priyanto; Rachmat, Agus; Elyazar, Iqbal F; Sukowati, Supratman

    2008-06-01

    Land use patterns and the occurrence of Anopheles species larvae were studied in Sukabumi District, West Java, Indonesia, from October 2004 to September 2005. Two land use maps derived using remote sensing were used. One map derived from Quickbird satellite images of 150 km2 of the Simpenan and Ciemas subdistricts (106 degrees 27' 53"-106 degrees 38' 38" E and 6 degrees 59' 59"-7 degrees 8' 46" S) in Sukabumi and one using ASTER images covering 4,000 km2 of Sukabumi District from 106 degrees 22' 15"-107 degrees 4' 1" E and 6 degrees 42' 50" - 7 degrees 26' 13" S. There was a total of 11 Anopheles spp. collected from 209 sampling locations in the area covered by the Quickbird image and a total of 15 Anopheles spp. collected from 1,600 sampling locations in the area covered by the ASTER map. For the area covered by the land use maps, ten species were found to have statistically positive relationships between land use class and species presence: Anopheles aconitus, An. annularis, An. barbirostris. An. flavirostris, An. insulaeflorum, An. kochi, An. maculatus, An. subpictus, An. sundaicus, and An. vagus. Quickbird and ASTER satellite images both produced land maps that were adequate for predicting species presence in an area. The land use classes associated with malaria vector breeding were rice paddy (An. aconitus, An. subpictus), plantation located near or adjacent to human settlements (An. maculatus), bush/shrub (An. aconitus, An. maculatus, An. sundaicus), bare land, and water body land use on the coast located < or = 250 m of the beach (An. sundaicus). Understanding the associations of habitat and species in one area, predictions of species presence or absence can be made prior to a ground survey allowing for accurate vector survey and control planning.

  6. 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).

    Science.gov (United States)

    Govindarajan, Marimuthu; Rajeswary, Mohan; Hoti, S L; Benelli, Giovanni

    2016-02-01

    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 84.93 μg/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 43.27 μg/ml, respectively) followed by An. subpictus (LC50=24.06 and 47.73 μg/ml), Cx. quinquefasciatus (LC50=26.08 and 52.19 μg/ml) and Cx. tritaeniorhynchus (LC50=27.95 and 54.87 μg/ml). Overall, this research adds knowledge to develop newer and safer natural larvicides against malaria, filariasis and Japanese encephalitis mosquito vectors. PMID:26850541

  7. A heterodimeric complex of the LRR proteins LRIM1 and APL1C regulates complement-like immunity in Anopheles gambiae

    Energy Technology Data Exchange (ETDEWEB)

    Baxter, Richard H.G.; Steinert, Stefanie; Chelliah, Yogarany; Volohonsky, Gloria; Levashina, Elena A.; Deisenhofer, Johann (CNRS-UMR); (UTSMC)

    2012-01-20

    The leucine-rich repeat (LRR) proteins LRIM1 and APL1C control the function of the complement-like protein TEP1 in Anopheles mosquitoes. The molecular structure of LRIM1 and APL1C and the basis of their interaction with TEP1 represent a new type of innate immune complex. The LRIM1/APL1C complex specifically binds and solubilizes a cleaved form of TEP1 without an intact thioester bond. The LRIM1 and APL1C LRR domains have a large radius of curvature, glycosylated concave face, and a novel C-terminal capping motif. The LRIM1/APL1C complex is a heterodimer with a single intermolecular disulfide bond. The structure of the LRIM1/APL1C heterodimer reveals an interface between the two LRR domains and an extensive C-terminal coiled-coil domain. We propose that a cleaved form of TEP1 may act as a convertase for activation of other TEP1 molecules and that the LRIM1/APL1C heterodimer regulates formation of this TEP1 convertase.

  8. Salivary Gland Proteome during Adult Development and after Blood Feeding of Female Anopheles dissidens Mosquitoes (Diptera: Culicidae)

    Science.gov (United States)

    Phattanawiboon, Benjarat; Jariyapan, Narissara; Mano, Chonlada; Roytrakul, Sittiruk; Paemanee, Atchara; Sor-Suwan, Sriwatapron; Sriwichai, Patchara; Saeung, Atiporn; Bates, Paul A.

    2016-01-01

    Understanding changes in mosquito salivary proteins during the time that sporozoite maturation occurs and after blood feeding may give information regarding the roles of salivary proteins during the malarial transmission. Anopheles dissidens (formerly Anopheles barbirostris species A1) is a potential vector of Plasmodium vivax in Thailand. In this study, analyses of the proteomic profiles of female An. dissidens salivary glands during adult development and after blood feeding were carried out using two-dimensional gel electrophoresis coupled with nano-liquid chromatography-mass spectrometry. Results showed at least 17 major salivary gland proteins present from day one to day 21 post emergence at 8 different time points sampled. Although there was variation observed, the patterns of protein expression could be placed into one of four groups. Fifteen protein spots showed significant depletion after blood feeding with the percentages of the amount of depletion ranging from 8.5% to 68.11%. The overall results identified various proteins, including a putative mucin-like protein, an anti-platelet protein, a long form D7 salivary protein, a putative gVAG protein precursor, a D7-related 3.2 protein, gSG7 salivary proteins, and a gSG6 protein. These results allow better understanding of the changes of the salivary proteins during the adult mosquito development. They also provide candidate proteins to investigate any possible link or not between sporozoite maturation, or survival of skin stage sporozoites, and salivary proteins. PMID:27669021

  9. Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia pholidotus (Diptera: Culicidae from Colombia

    Directory of Open Access Journals (Sweden)

    Jesús Eduardo Escovar

    2014-07-01

    Full Text Available Since 1984, Anopheles (Kerteszia lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

  10. [Malaria mosquitoes (Diptera, Culicidae, Anopheles) of North Tajikistan, their ecology, and role in the transmission of malaria pathogens].

    Science.gov (United States)

    Kadamov, D S; Zvantseva, A B; Karimov, S S; Gordeev, M I; Goriacheva, I I; Ezhov, M N; Tadzhiboev, A

    2012-01-01

    Five species of malaria mosquitoes: An. artemievi, An. claviger, An. hyrcanus, An. superpictus, and An. pulcherrimus were found in North Tajikistan in 2006 - 2007. Species affiliation was identified according to the morphological signs of their larvae and imagoes, and by using the polymerase chain reaction-restriction fragment length polymorphism analysis. There was a larger number of An. hyrcanus (34%), An. artemievi (29%), and An. pulcherrimus (24%) and a smaller number of An. superpictus (11%); and An. claviger was few (2%). The hatching sites of the above species and the preferred types of their day refuges were found. The intensity of attack of different Anopheles species on humans and animals was studied. Among the North Tajikistan malaria mosquitoes, An. pulcherrimus and An. superpictus are of the greatest epidemiological importance as vehicles for transmission of malaria pathogens. An. artemievi and An. hyrcanus are minor vehicles. At present, An. claviger is of no epidemiological significance in transmitting malaria in North Tajikistan.

  11. Mosquito adulticidal and repellent activities of botanical extracts against malarial vector,Anopheles stephensi Liston (Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    Marimuthu Govindarajan; Rajamohan Sivakumar

    2011-01-01

    Objective:To determine the adulticidal and repellent activities of different solvent leaf extracts ofEclipta alba (E. alba) andAndrographis paniculata (A. paniculata)against malarial vector, Anopheles stephensi (An. stephensi).Methods:Adulticidal efficacy of the crude leaf extracts ofE. alba andA. paniculata with five different solvents like benzene, hexane, ethyl acetate, methanol and chloroform was tested against the five to six day old adult female mosquitoes of An. stephensi. The adult mortality was observed after24 h under the laboratory conditions. The repellent efficacy was determined againstAn. stephensimosquito species at three concentrations viz.,1.0, 2.5 and 5.0 mg/cm2 under laboratory conditions.Results: Among the tested solvents the maximum efficacy was observed in the methanol extract. TheLC50 andLC90 values ofE. alba andA. paniculata against adults ofAn. stephensiwere150.36, 130.19 ppm and285.22, 244.16ppm, respectively. No mortality was observed in controls. Thechi-square values were significant at P<0.05 level. Methanol extract of E. alba andA. paniculata was produce maximum repellency againstAn. stephensi.Conclusions:From the results it can be concluded the crude extract ofE. alba andA. paniculatawas an excellent potential for controllingAn. stephensimosquitoes.

  12. Deforestation and Malaria on the Amazon Frontier: Larval Clustering of Anopheles darlingi (Diptera: Culicidae) Determines Focal Distribution of Malaria.

    Science.gov (United States)

    Barros, Fábio S M; Honório, Nildimar A

    2015-11-01

    We performed bimonthly mosquito larval collections during 1 year, in an agricultural settlement in the Brazilian Amazon, as well as an analysis of malaria incidence in neighboring houses. Water collections located at forest fringes were more commonly positive for Anopheles darlingi larvae and Kulldorff spatial analysis pinpointed significant larval clusters at sites directly beneath forest fringes, which were called larval "hotspots." Remote sensing identified 43 "potential" hotspots. Sampling of these areas revealed an 85.7% positivity rate for A. darlingi larvae. Malaria was correlated with shorter distances to potential hotpots and settlers living within 400 m of potential hotspots had a 2.60 higher risk of malaria. Recently arrived settlers, usually located closer to the tip of the triangularly shaped deforestation imprints of side roads, may be more exposed to malaria due to their proximity to the forest fringe. As deforestation progresses, transmission decreases. However, forest remnants inside deforested areas conferred an increased risk of malaria. We propose a model for explaining frontier malaria in the Amazon: because of adaptation of A. darlingi to the forest fringe ecotone, humans are exposed to an increased transmission risk when in proximity to these areas, especially when small dams are created on naturally running water collections. PMID:26416110

  13. Green chemistry focus on optimization of silver nanoparticles using response surface methodology (RSM) and mosquitocidal activity: Anopheles stephensi (Diptera: Culicidae).

    Science.gov (United States)

    Ondari Nyakundi, Erick; Padmanabhan, M Nalini

    2015-01-01

    There is an exigent necessity for development of environmental friendly bio-control agent(s) for elimination of mosquito due to increased resistance resurgence against synthetic control agents. Mosquito control strategy will lay a strong foundation to malaria exclusion or it can be curbed to certain level especially in the developing nations. In this study, silver nanoparticles were synthesized by green chemistry approach using Tridax procumbens leaf extract as a reducing agent. The reaction medium involved in the synthesis process was optimized by statistical experimental design using response surface methodology to obtain better yield, uniform size, shape and stability. Further, these synthesized nanoparticles were confirmed through UV-Visible, FT-IR spectroscopy, PSA and SEM Subsequently, the bioefficacy of these particles were investigated on Anopheles stephensi for larvicidal and pupicidal activity. Interestingly, time period of 90 min, temperature of 76±2 °C, pH 7.2±2, 2 mM silver nitrate (AgNO3), 3mM PEG and 2mM PVP showed excellent parameters for bioprocess design for large scale production of stabilized nanoparticles. A concentration of 5 ppm of PVP stabilized nanoparticles exhibited 100% mortality. Thus, the obtained results clearly suggest that silver nanoparticles stabilized by PEG and PVP may have important function as stabilizers, dispersants as well as larvicides for mosquito control.

  14. Correlation of survival rates of Anopheles dirus A (Diptera: Culicidae) with different infection densities of Plasmodium cynomolgi*

    Science.gov (United States)

    Klein, T. A.; Harrison, B. A.; Grove, J. S.; Dixon, S. V.; Andre, R. G.

    1986-01-01

    The survival rates are described for 36 paired replicates of Anopheles dirus A mosquitos that had been allowed to engorge themselves on rhesus monkeys that were either infected or non-infected with Plasmodium cynomolgi. The survival rates of infected mosquitos with a mean number of oocysts less than 10 did not differ significantly from those that were non-infected; however, there was a significant difference in the survival rates of non-infected groups and those with a mean number of oocysts in the range 10-40, 41-70, or ≥71. Daily survival rates for non-infected and infected mosquitos did not differ significantly during the first 8 days of extrinsic incubation. In contrast, for the period 9-30 days the survival rates of mosquitos with mean number of oocysts ≥41 were significantly different from those of non-infected mosquitos. The cumulative daily survival rates of mosquitos with mean number of oocysts up to 40, 41-70, or ≥71 decreased with the oocyst count. Mosquitos with a mean number of oocysts ≥71 frequently exhibited excessive numbers of bacteria and deterioration of both their guts and salivary glands. PMID:3493859

  15. STATUS OF INSECTICIDE RESISTANCE IN ANOPHELES CULICIFACIES (DIPTERA: CULICIDAE IN GHASREGHAND DISTRICT, SISTAN AND BALUCHISTAN PROVINCE, IRAN, (1997

    Directory of Open Access Journals (Sweden)

    H. Vatandoost

    1999-08-01

    Full Text Available Anopheles culicifacies s.l. plays an important role In transmission of malaria in Sistan and Baluchistan province, southeastern Iran. Adult susceptibility test on fieltt-collccled mosquitoes was conducted in Ohasreghand district. WHO diagnostic test procedures revealed that adult females were resistant to 0.4% dieUirin (mortality 64.5 ± 3.13, tolerant to 0.1% propoxur (mortality 88.5 ± 2.24 and susceptible to 4% DDT (mortality 98.75 ± 0.8. 5% malathion (mortality 100%, 0.1% bendiocarb (mortality 98.86 ± 0.7, 0.25% pcrmcthrin (mortality 98.4 ± 0.1, ami 0.1% lamhdacyhalothrin (mortality 100%. Malathion and lamhdacyhalothrin had the highest efficacy against this species when they were exposed at the diagnostic dose for 1 hour followed by a 24 hour recovery period. DieUirin, DDT a nil malathion had been used for malaria control as an indoor residual spraying. Tlic implication of these findings in the control programme is discussed.

  16. A molecular phylogeny of Anopheles annulipes (Diptera: Culicidae) sensu lato: the most species-rich anopheline complex.

    Science.gov (United States)

    Foley, D H; Wilkerson, R C; Cooper, R D; Volovsek, M E; Bryan, J H

    2007-04-01

    The Australasian Annulipes Complex is the most species-rich among Anopheles mosquitoes, with at least 15 sibling species suspected. Members of this complex are the most likely vectors of malaria in the past in southern Australia and are involved in the spread of myxomatosis among rabbits. In this, the first comprehensive molecular study of the Annulipes Complex, 23 ITS2 rDNA variants were detected from collections throughout Australia and Papua New Guinea, including diagnostic variants for the previously identified An. annulipes species A-G. Specimens of each ITS2 variant were sequenced for portions of the mitochondrial COI, COII and nuclear EF-1alpha genes. Partitioned Bayesian and Maximum Parsimony analyses confirmed the monophyly of the Annulipes Complex and revealed at least 17 clades that we designate species A-Q. These species belong to two major clades, one in the north and one mainly in the south, suggesting that climate was a driver of species radiation. We found that 65% (11) of the 17 sibling species recorded here had unique COI sequences, suggesting that DNA barcoding will be useful for diagnosing species within the Annulipes Complex. A comparison of the taxa revealed morphological characters that may be diagnostic for some species. Our results substantially increase the size of the subgenus Cellia in Australasia, and will assist species-level studies of the Annulipes Complex. PMID:17126567

  17. Green chemistry focus on optimization of silver nanoparticles using response surface methodology (RSM) and mosquitocidal activity: Anopheles stephensi (Diptera: Culicidae).

    Science.gov (United States)

    Ondari Nyakundi, Erick; Padmanabhan, M Nalini

    2015-01-01

    There is an exigent necessity for development of environmental friendly bio-control agent(s) for elimination of mosquito due to increased resistance resurgence against synthetic control agents. Mosquito control strategy will lay a strong foundation to malaria exclusion or it can be curbed to certain level especially in the developing nations. In this study, silver nanoparticles were synthesized by green chemistry approach using Tridax procumbens leaf extract as a reducing agent. The reaction medium involved in the synthesis process was optimized by statistical experimental design using response surface methodology to obtain better yield, uniform size, shape and stability. Further, these synthesized nanoparticles were confirmed through UV-Visible, FT-IR spectroscopy, PSA and SEM Subsequently, the bioefficacy of these particles were investigated on Anopheles stephensi for larvicidal and pupicidal activity. Interestingly, time period of 90 min, temperature of 76±2 °C, pH 7.2±2, 2 mM silver nitrate (AgNO3), 3mM PEG and 2mM PVP showed excellent parameters for bioprocess design for large scale production of stabilized nanoparticles. A concentration of 5 ppm of PVP stabilized nanoparticles exhibited 100% mortality. Thus, the obtained results clearly suggest that silver nanoparticles stabilized by PEG and PVP may have important function as stabilizers, dispersants as well as larvicides for mosquito control. PMID:26010566

  18. Monoclonal antibodies AC-43 and AC-29 disrupt Plasmodium vivax development in the Indian malaria vector Anopheles culicifacies (Diptera: culicidae)

    Indian Academy of Sciences (India)

    Manoj Chugh; B R Gulati; S K Gakhar

    2010-03-01

    A repertoire of monoclonal antibodies (mAbs) was generated against the midgut proteins of Anopheles culicifacies mosquitoes. The mAbs AC-43 and AC-29 significantly inhibited Plasmodium vivax development inside the mosquito midgut. The number of oocysts that developed was reduced by 78.6% when mosquitoes ingested a combination of these two mAbs along with the blood meal. AC-43 mAb binds to the epitope common in 97, 80 and 43 kDa polypeptides from the midgut protein extract, as indicated by western blot analysis. Similarly, the mAb AC-29 recognized 52, 44, 40 and 29 kDa polypeptides. These female midgut-specific polypeptides are shared between An. culicifacies and An. stephensi, two major vectors of malaria in India. Deglycosylation assays revealed that -linked carbohydrates are the major components in epitopes corresponding to AC-43 and AC-29. Gold particle labelling revealed that both these mAbs preferentially bind to glycoproteins at the apical microvilli and the microvillus-associated network present inside transverse sections of the gut epithelium. These regions are particularly known to have receptors for ookinetes, which enable them to cross this epithelial barrier and provide them with certain necessary chemicals or components for further development into oocysts. Therefore, these glycoproteins appear to be potential candidates for a vectordirected transmission-blocking vaccine (TBV).

  19. Using remote sensing to map larval and adult populations of Anopheles hyrcanus (Diptera: Culicidae a potential malaria vector in Southern France

    Directory of Open Access Journals (Sweden)

    Roger François

    2008-02-01

    Full Text Available Abstract Background Although malaria disappeared from southern France more than 60 years ago, suspicions of recent autochthonous transmission in the French Mediterranean coast support the idea that the area could still be subject to malaria transmission. The main potential vector of malaria in the Camargue area, the largest river delta in southern France, is the mosquito Anopheles hyrcanus (Diptera: Culicidae. In the context of recent climatic and landscape changes, the evaluation of the risk of emergence or re-emergence of such a major disease is of great importance in Europe. When assessing the risk of emergence of vector-borne diseases, it is crucial to be able to characterize the arthropod vector's spatial distribution. Given that remote sensing techniques can describe some of the environmental parameters which drive this distribution, satellite imagery or aerial photographs could be used for vector mapping. Results In this study, we propose a method to map larval and adult populations of An. hyrcanus based on environmental indices derived from high spatial resolution imagery. The analysis of the link between entomological field data on An. hyrcanus larvae and environmental indices (biotopes, distance to the nearest main productive breeding sites of this species i.e., rice fields led to the definition of a larval index, defined as the probability of observing An. hyrcanus larvae in a given site at least once over a year. Independent accuracy assessments showed a good agreement between observed and predicted values (sensitivity and specificity of the logistic regression model being 0.76 and 0.78, respectively. An adult index was derived from the larval index by averaging the larval index within a buffer around the trap location. This index was highly correlated with observed adult abundance values (Pearson r = 0.97, p An. hyrcanus larval and adult populations from the landscape indices. Conclusion This work shows that it is possible to use

  20. Infections expérimentales de Anopheles gambiae avec différentes souches de Plasmodium falciparum issu de porteurs de gamétocytes naturellement infectés au Cameroun : facteurs influençant l'infection des moustiques

    OpenAIRE

    Tchuinkam, T.; B. Mulder; K. Dechering; Stoffels, H.; Verhave, J P; Cot, Michel; Carnevale, Pierre; Meuwissen, J. H. E. T.; Robert, Vincent

    1994-01-01

    L'infectivité des porteurs de gamétocytes de #Plasmodium falciparum$ présentant des symptômes palustres dans la ville de Yaoundé pour une souche d'Anopheles gambiae a été évaluée et les facteurs susceptibles d'influencer la réussite des infections expérimentales ont été recherchés. 139 infections expérimentales avec le sang de différents porteurs de gamétocytes ont été effectuées. Après dissection d'au moins 20 moustiques par expérimentation, 86 (62%) porteurs de gamétocytes ont donné au moin...

  1. DNA Barcodes indicate members of the Anopheles fluviatilis (Diptera: Culicidae) species complex to be conspecific in India.

    Science.gov (United States)

    Pradeep Kumar, N; Krishnamoorthy, N; Sahu, S S; Rajavel, A R; Sabesan, S; Jambulingam, P

    2013-05-01

    Anopheles fluviatilis, a major vector of malaria in India has been described as a complex of three sibling species members, named as S, T and U, based on variations in chromosomal inversions. Also, ribosomal DNA markers (repetitive Internal Transcribed Spacer 2 (ITS2) and 28S D3 region) were described to differentiate these three sibling species members. However, controversies prevail on the genetic isolation status of these cryptic species. Hence, we evaluated this taxonomic incongruence employing DNA barcoding, the well established methodology for species identification, using 60 An. fluviatilis sensu lato specimens, collected from two malaria endemic eastern states of India. These specimens were also subjected to sibling species characterization by ITS2 and D3 DNA markers. The former marker identified 31 specimens among these as An. fluviatilis S and 21 as An. fluviatilis T. Eight specimens amplified DNA fragments specific for both S and T. The D3 marker characterized 39 specimens belonging to species S and 21 to species T. Neither marker identified species U. Neighbor Joining analysis of mitochondrial cytochrome c oxidase gene 1 sequences (the DNA barcode) categorized all the 60 specimens into a single operational taxonomic unit, their Kimura 2 parameter (K2P) genetic variability being only 0.8%. The genetic differentiation (FST ) and gene flow (Nm ) estimates were 0.00799 and 62.07, respectively, indicating these two 'species' (S & T) as genetically con-specific intermixing populations with negligible genetic differentiation. Earlier investigations have refuted the existence of species U. Also, this study demonstrated that An. fluviatilis and the closely related An. minimus could be taxonomically differentiated by the DNA Barcode approach (K2P = 5.0%). PMID:23398631

  2. Larvicidal and repellent properties of Adansonia digitata against medically important human malarial vector mosquito Anopheles stephensi (Diptera: Culicidae

    Directory of Open Access Journals (Sweden)

    K. Krishnappa , K. Elumalai , S. Dhanasekaran & J. Gokulakrishnan

    2012-06-01

    Full Text Available Background & objectives: Development of plant-based alternative compounds for mosquito control has gainedimportance now-a-days, in view of increasing resistance in mosquito vectors to existing insecticides. The larvicidaland repellent activities of benzene, chloroform, hexane and methanol leaf extracts of Indian medicinal plant,Adansonia digitata were investigated against malarial vector, Anopheles stephensi.Methods: In all, 25 III instar larvae of An. stephensi were exposed to various concentrations (30–180 mg/l in thelaboratory by using the standard protocol described by WHO (2005. The larvae were exposed for 24 h andmortalities were subjected to log-probit analysis. Repellent activity of crude leaf extract at the dosages of 2, 4and 6 mg/cm2 was evaluated in a net cage (45 × 30 × 45 cm containing 100 blood starved female mosquitoes ofAn. stephensi using the protocol of WHO (1996.Results: Preliminary phytochemical analysis of A. digitata showed the presence of triterpenoids and saponins.The LC50 and LC90 values of hexane, benzene, chloroform, and methanol extracts of A. digitata against An.stephensi larvae in 24 h were 111.32, 97.13, 88.55, 78.18 and 178.63, 176.19, 168.14, 155.42 mg/l, respectively.The repellent activity of methanol extract was found to be most effective and at higher concentration of 6mg/cm2 benzene, chloroform hexane and methanol extracts provided 100% protection up to 150, 180, 120 and210 min against An. stephensi, respectively.Conclusion: The preliminary study indicated that A. digitata showed larvicidal and repellent activities againstAn. stephensi and could be used for controlling mosquitoes. Further studies are indicated to purify the activecompounds from these plants for developing larvicide and repellents.

  3. Baseline Susceptibility of Different Geographical Strains of Anopheles stephensi (Diptera: Culicidae to Temephos in Malarious Areas of Irana.

    Directory of Open Access Journals (Sweden)

    Aboozar Soltani

    2013-06-01

    Full Text Available Malaria still remains a public health problem in Iran. There are different vector control interventions such as insecticide spraying. The present study was carried out to determine the susceptibility status of Anopheles stephensi larvae to temephos as a national plan for monitoring and mapping of insecticide resistance.Eight different localities in two main malarious provinces were determined as field collecting sites. Mosquitoes were collected from the field and reared in an insectray. Susceptibility assays were carried out according to the WHO method. The laboratory reared susceptible Beech-Lab strain was used for comparison. Data were analyzed using Probit analysis to determine LC50 and LC90 values.Susceptibility of An. stephensi to temephos indicated that the LC50 ranged from 0.0022 mg/l to 0.0141 mg/l. Although all field strains were susceptible to temephos, considerable variations in temephos resistance ratios of field strains were noticed from all the localities studied in comparison with the susceptible strain. A low level of resistance ratio was noticed in An. stephensi populations except for the Chabahar strain (RR= 4.27 fold. All field-collected An. stephensi populations exhibited homogeneity to the larvicide except for Bandar Abbas and Hormoodar village strains (P> 0.05%.Due to intensive use of temephos in the neighboring countries and occurrence of resistant to this insecticide in the main malaria vector in the region, insecticide resistance gene may evolve in the populations of An. stephensi. If temephos be applied as a larvicide it should be used judiciously for resistance management, as rotation strategy.

  4. Baseline Susceptibility of Different Geographical Strains of Anopheles stephensi (Diptera: Culicidae to Temephos in Malarious Areas of Iran

    Directory of Open Access Journals (Sweden)

    Aboozar Soltani

    2013-03-01

    Full Text Available Background: Malaria still remains a public health problem in Iran. There are different vector control interventions such as insecticide spraying. The present study was carried out to determine the susceptibility status of Anopheles stephensi larvae to temephos as a national plan for monitoring and mapping of insecticide resistanceMethods: Eight different localities in two main malarious provinces were determined as field collecting sites. Mos­quitoes were collected from the field and reared in an insectray. Susceptibility assays were carried out according to the WHO method. The laboratory reared susceptible Beech-Lab strain was used for comparison. Data were analyzed using Probit analysis to determine LC50 and LC90 values.Results: Susceptibility of An. stephensi to temephos indicated that the LC50 ranged from 0.0022 mg/l to 0.0141 mg/l. Although all field strains were susceptible to temephos, considerable variations in temephos resistance ratios of field strains were noticed from all the localities studied in comparison with the susceptible strain. A low level of resistance ratio was noticed in An. stephensi populations except for the Chabahar strain (RR= 4.27 fold. All field-collected An. stephensi populations exhibited homogeneity to the larvicide except for Bandar Abbas and Hormoodar village strains (P> 0.05%.Conclusion: Due to intensive use of temephos in the neighboring countries and occurrence of resistant to this insec­ticide in the main malaria vector in the region, insecticide resistance gene may evolve in the populations of An. stephensi. If temephos be applied as a larvicide it should be used judiciously for resistance management, as rotation strategy

  5. Present habitat suitability for Anopheles atroparvus (Diptera, Culicidae and its coincidence with former malaria areas in mainland Portugal

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    César Capinha

    2009-05-01

    Full Text Available Malaria was a major health problem in the first half of the 20th Century in mainland Portugal. Nowadays, although the disease is no longer endemic, there is still the risk of future endemic infections due to the continuous occurrence of imported cases and the possibility of transmission in the country by Anopheles atroparvus Van Thiel, 1927. Since vector abundance constitute one of the foremost factors in malaria transmission, we have created several habitat suitability models to describe this vector species’ current distribution. Three different correlative models; namely (i a multilayer perceptron artificial neural network (MLP-ANN; (ii binary logistic regression (BLR; and (iii Mahalanobis distance were used to combine the species records with a set of five environmental predictors. Kappa coefficient values from k-fold cross-validation records showed that binary logistic regression produced the best predictions, while the other two models also produced acceptable results. Therefore, in order to reduce uncertainty, the three suitability models were combined. The resulting model identified high suitability for An. atroparvus in the majority of the country with exception of the northern and central coastal areas. Malaria distribution during the last endemic period in the country was also compared with the combined suitability model, and a high degree of spatial agreement was obtained (kappa = 0.62. It was concluded that habitat suitability for malaria vectors can constitute valuable information on the assessment of several spatial attributes of the disease. In addition, the results suggest that the spatial distribution of An. atroparvus in the country remains very similar to the one known about seven decades ago.

  6. Population genetic structure of the major malaria vector Anopheles darlingi (Diptera: Culicidae from the Brazilian Amazon, using microsatellite markers

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    Vera Margarete Scarpassa

    2007-06-01

    Full Text Available The population genetic structure of Anopheles darlingi, the major human malaria vector in the Neotropics, was examined using seven microsatellite loci from nine localities in central and western Amazonian Brazil. High levels of genetic variability were detected (5-25 alleles per locus; H E = 0.519-0.949. There was deviation from Hardy-Weinberg Equilibrium for 59.79% of the tests due to heterozygote deficits, while the analysis of linkage disequilibrium was significant for only two of 189 (1.05% tests, most likely caused by null alleles. Genetic differentiation (F ST = 0.001-0.095; Nm = 4.7-363.8 indicates that gene flow is extensive among locations < 152 km apart (with two exceptions and reduced, but not absent, at a larger geographic scale. Genetic and geographic distances were significantly correlated (R² = 0.893, P < 0.0002, supporting the isolation by distance (IBD model. The overall estimate of Ne was 202.4 individuals under the linkage disequilibrium model, and 8 under the heterozygote excess model. Analysis of molecular variance showed that nearly all variation (~ 94% was within sample locations. The UPGMA phenogram clustered the samples geographically, with one branch including 5/6 of the state of Amazonas localities and the other branch the Acre, Rondônia, and remaining Amazonas localities. Taken together, these data suggest little genetic structure for An. darlingi from central and western Amazonian Brazil. These findings also imply that the IBD model explains nearly all of the differentiation detected. In practical terms, populations of An. darlingi at distances < 152 km should respond similarly to vector control measures, because of high gene flow.

  7. Expression Profiles and RNAi Silencing of Inhibitor of Apoptosis Transcripts in Aedes, Anopheles, and Culex Mosquitoes (Diptera: Culicidae).

    Science.gov (United States)

    Puglise, Jason M; Estep, Alden S; Becnel, James J

    2016-03-01

    Effective mosquito control is vital to curtail the devastating health effects of many vectored diseases. RNA interference (RNAi)-mediated control of mosquitoes is an attractive alternative to conventional chemical pesticides. Previous studies have suggested that transcripts for inhibitors of apoptosis (IAPs) may be good RNAi targets. To revisit and extend previous reports, we examined the expression of Aedes aegypti (L.) IAPs (AaeIAPs) 1, 2, 5, 6, 9, and a viral IAP-associated factor (vIAF) as well as Anopheles quadrimaculatus Say and Culex quinquefasciatus Say IAP1 homologs (AquIAP1 and CquIAP1) in adult females. Expression profiles of IAPs suggested that some older female mosquitoes had significantly higher IAP mRNA levels when compared to the youngest ones. Minor differences in expression of AaeIAPs were observed in mosquitoes that imbibed a bloodmeal, but the majority of the time points (up to 48 h) were not significantly different. Although in vitro experiments with the Ae. aegypti Aag-2 cell line demonstrated that the various AaeIAPs could be effectively knocked down within one day after dsRNA treatment, only Aag-2 cells treated with dsIAP1 displayed apoptotic morphology. Gene silencing and mortality were also evaluated after topical application and microinjection of the same dsRNAs into female Ae. aegypti. In contrast to previous reports, topical administration of dsRNA against AaeIAP1 did not yield a significant reduction in gene expression or increased mortality. Knockdown of IAP1 and other IAPs by microinjection did not result in significant mortality. In toto, our findings suggest that IAPs may not be suitable RNAi targets for controlling adult mosquito populations.

  8. Ovicidal and repellent activities of botanical extracts against Culex quinquefasciatus,Aedes aegypti and Anopheles stephensi(Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    Govindarajan; M; Mathivanan; T; Elumalai; K; Krishnappa; K; Anandan; A

    2011-01-01

    Objective:To determine the ovicidal and repellent activities of methanol leaf extract of Ervatamia coronaria(E.coronaria) and Caeslpinia pulckerrima(C.pulcherrima) against Culex quinquefasciatus(Cx.quinquefasciatus),Aedes aegypti(Ae.aegypti) and Anopheles stephensi(An. stephensi).Methods:The ovicidal activity was determined against three mosquito species at various concentrations ranging from 50-450 ppm under the laboratory conditions.The hatch rates were assessed 48 h after treatment.The repellent efficacy was determined against three mosquito species at three concentrations viz.,1.0,2.5 and 5.0 mg/cm under the laboratory conditions. Results:The crude extract of E.coronaria exerted zero hatchability(100%mortality) at 250.200 and 150 ppm for Cx.quinqitefasciatus,Ae.aegypti and An.stephensi,respectively.The crude extract of C.pulchenima exerted zero hatchability(100%mortality) at 375.300 and 225 ppm for Cx.quinquefasciatus,Ae.aegypti and An.Stephensi,respectively.The methanol extract of E. coronaria found to be more repellenct than C.pukherrima extract.A higher concentration of 5.0 mg/cm~2 provided 100%protection up to 150.180 and 210 min against Cx.quinquefasciatus,Ae. aegypti and An.stephensi,respectively.The results clearly showed that repellent activity was dose dependent.Conclusions:From the results it can be concluded the crude extracts of E.coronaria and C.pukherrima are an excellent potential for controlling Cx.quinquefasciatus,Ae.aegypti and An.stephensi mosquitoes.

  9. Survival of immature Anopheles arabiensis (Diptera: Culicidae in aquatic habitats in Mwea rice irrigation scheme, central Kenya

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    Jacob Benjamin

    2006-11-01

    Full Text Available Abstract Background The survivorship and distribution of Anopheles arabiensis larvae and pupae was examined in a rice agro-ecosystem in Mwea Irrigation Scheme, central Kenya, from August 2005 to April 2006, prior to implementation of larval control programme. Methods Horizontal life tables were constructed for immatures in semi-field condition. The time spent in the various immature stages was determined and survival established. Vertical life tables were obtained from five paddies sampled by standard dipping technique. Results Pre-adult developmental time for An. arabiensis in the trays in the experimental set up in the screen house was 11.85 days from eclosion to emergence. The mean duration of each instar stage was estimated to be 1.40 days for first instars, 2.90 days for second instars, 1.85 days for third instars, 3.80 days for fourth instars and 1.90 days for pupae. A total of 590 individuals emerged into adults, giving an overall survivorship from L1 to adult emergence of 69.4%. A total of 4,956 An. arabiensis immatures were collected in 1,400 dips throughout the sampling period. Of these, 55.9% were collected during the tillering stage, 42.5% during the transplanting period and 1.6% during the land preparation stage. There was a significant difference in the An. arabiensis larval densities among the five stages. Also there was significant variation in immature stage composition for each day's collection in each paddy. These results indicate that the survival of the immatures was higher in some paddies than others. The mortality rate during the transplanting was 99.9% and at tillering was 96.6%, while the overall mortality was 98.3%. Conclusion The survival of An. arabiensis immatures was better during the tillering stage of rice growth. Further the survival of immatures in rice fields is influenced by the rice agronomic activities including addition of nitrogenous fertilizers and pesticides. For effective integrated vector management

  10. A Physical Map for an Asian Malaria Mosquito, Anopheles stephensi

    OpenAIRE

    Maria V Sharakhova; Xia, Ai; Tu, Zhijian; Shouche, Yogesh S.; Unger, Maria F; Sharakhov, Igor V

    2010-01-01

    Physical mapping is a useful approach for studying genome organization and evolution as well as for genome sequence assembly. The availability of polytene chromosomes in malaria mosquitoes provides a unique opportunity to develop high-resolution physical maps. We report a 0.6-Mb-resolution physical map consisting of 422 DNA markers hybridized to 379 chromosomal sites of the Anopheles stephensi polytene chromosomes. This makes An. stephensi second only to Anopheles gambiae in density of a phys...

  11. Nigeria Anopheles vector database: an overview of 100 years' research.

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    Patricia Nkem Okorie

    Full Text Available Anopheles mosquitoes are important vectors of malaria and lymphatic filariasis (LF, which are major public health diseases in Nigeria. Malaria is caused by infection with a protozoan parasite of the genus Plasmodium and LF by the parasitic worm Wuchereria bancrofti. Updating our knowledge of the Anopheles species is vital in planning and implementing evidence based vector control programs. To present a comprehensive report on the spatial distribution and composition of these vectors, all published data available were collated into a database. Details recorded for each source were the locality, latitude/longitude, time/period of study, species, abundance, sampling/collection methods, morphological and molecular species identification methods, insecticide resistance status, including evidence of the kdr allele, and P. falciparum sporozoite rate and W. bancrofti microfilaria prevalence. This collation resulted in a total of 110 publications, encompassing 484,747 Anopheles mosquitoes in 632 spatially unique descriptions at 142 georeferenced locations being identified across Nigeria from 1900 to 2010. Overall, the highest number of vector species reported included An. gambiae complex (65.2%, An. funestus complex (17.3%, An. gambiae s.s. (6.5%. An. arabiensis (5.0% and An. funestus s.s. (2.5%, with the molecular forms An. gambiae M and S identified at 120 locations. A variety of sampling/collection and species identification methods were used with an increase in molecular techniques in recent decades. Insecticide resistance to pyrethroids and organochlorines was found in the main Anopheles species across 45 locations. Presence of P. falciparum and W. bancrofti varied between species with the highest sporozoite rates found in An. gambiae s.s, An. funestus s.s. and An. moucheti, and the highest microfilaria prevalence in An. gambiae s.l., An. arabiensis, and An. gambiae s.s. This comprehensive geo-referenced database provides an essential baseline on

  12. Evaluation of Methods for Sampling the Malaria Vector Anopheles darlingi (Diptera, Culicidae) in Suriname and the Relation With Its Biting Behavior

    NARCIS (Netherlands)

    Hiwat-van Laar, H.; Rijk, de M.; Andriessen, R.; Koenraadt, C.J.M.; Takken, W.

    2011-01-01

    The effectiveness of CO2-baited and human-baited mosquito traps for the sampling of Anopheles darlingi Root was evaluated and compared with human landing collections in Suriname. Biting preferences of this mosquito on a human host were studied and related to trapping data. Traps used were the Center

  13. Studies on larvicidal and pupicidal activity of Leucas aspera Willd. (Lamiaceae) and bacterial insecticide, Bacillus sphaericus, against malarial vector, Anopheles stephensi Liston (Diptera: Culicidae)

    Science.gov (United States)

    The efficacy of whole plant ethanolic extracts of Leucas aspera and of Bacillus sphaericus was determined for larvae and pupae of Anopheles stephensi. When larvae were exposed to one of five concentrations of plant extract (6%, 8%, 10%, 12%, and 14%) for 24 h, mortality in 4th instars ranged from 1...

  14. Geometric morphometric analysis of Colombian Anopheles albimanus (Diptera: Culicidae) reveals significant effect of environmental factors on wing traits and presence of a metapopulation

    OpenAIRE

    Giovan F Gómez; Márquez, Edna J.; Gutiérrez, Lina A.; Conn, Jan E.; Correa, Margarita M.

    2014-01-01

    Anopheles albimanus is a major malaria mosquito vector in Colombia. In the present study, wing variability (size and shape) in An. albimanus populations from Colombian Maracaibo and Chocó bio-geographical eco-regions and the relationship of these phenotypic traits with environmental factors were evaluated. Microsatellite and morphometric data facilitated a comparison of the genetic and phenetic structure of this species. Wing size was influenced by elevation and relative humidity, whereas win...

  15. Larval habitat associations with human land uses, roads, rivers and land cover for Anopheles albimanus, An. pseudopunctipennis and An. punctimacula (Diptera: Culicidae in coastal and highland Ecuador

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    Lauren Lynn Pinault

    2012-03-01

    Full Text Available Larval habitat for three highland Anopheles species: Anopheles albimanus Wiedemann, Anopheles pseudopunctipennis Theobald, and Anopheles punctimacula Dyar & Knab was related to human land uses, rivers, roads, and remotely sensed land cover classifications in the western Ecuadorian Andes. Of the five commonly-observed human land uses, cattle pasture (n=30 provided potentially suitable habitat for An. punctimacula and An. albimanus in less than 14% of sites, and was related in a Principal Components Analysis (PCA to the presence of macrophyte vegetation, greater surface area, clarity and algae cover. Empty lots (n=30 were related in the PCA to incident sunlight and provided potential habitat for An. pseudopunctipennis and An. albimanus in less than 14% of sites. The other land uses surveyed (banana, sugarcane and mixed tree plantations; n=28, 21, 25, respectively provided very little standing water that could potentially be used for larval habitat. River edges and eddies (n=41 were associated with greater clarity, depth, temperature and algae cover, which provide potentially suitable habitat for An. albimanus in 58% of sites and An. pseudopunctipennis in 29% of sites. Road-associated water bodies (n=38 provided potential habitat for An. punctimacula in 44% of sites and An. albimanus in 26% of sites surveyed. Species collection localities were compared to land cover classifications using Geographic Information Systems software. All three mosquito species were associated more often with the category "closed/open broadleaved evergreen and/or semi-deciduous forests" than expected (P ≤ 0.01 in all cases, given such a habitat's abundance. This study provides evidence that specific human land uses create habitat for potential malaria vectors in highland regions of the Andes.

  16. Studies on mosquitoes (Diptera: Culicidae and anthropicenvironment: 5- Breeding of Anopheles albitarsis in flooded rice fields in South-Eastern Brazil

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    Forattini Oswaldo Paulo

    1994-01-01

    Full Text Available Studies on breeding Anopheles albitarsis and association with rice growth in irrigated paddy fields were carried out during the rice cultivation cycle from December 1993 to March 1994. This period corresponded to the length of time of permanent paddy flooding. Breeding occurred in the early stage up until five weeks after transplantation when rice plant height was small. That inverse correlation may give potential direction to control measures.

  17. Molecular species identification, host preference and detection of myxoma virus in the Anopheles maculipennis complex (Diptera: Culicidae) in southern England, UK.

    OpenAIRE

    Brugman, VA; Hernández-Triana, LM; Prosser, SW; Weland, C; Westcott, DG; Fooks, AR; Johnson, N

    2015-01-01

    Background Determining the host feeding patterns of mosquitoes by identifying the origin of their blood-meals is an important part of understanding the role of vector species in current and future disease transmission cycles. Collecting large numbers of blood-fed mosquitoes from the field is difficult, therefore it is important to maximise the information obtained from each specimen. This study aimed to use mosquito genome sequence to identify the species within Anopheles maculipennis sensu l...

  18. Studies on mosquitoes (Diptera: Culicidae and anthropicenvironment: 5- Breeding of Anopheles albitarsis in flooded rice fields in South-Eastern Brazil

    Directory of Open Access Journals (Sweden)

    Oswaldo Paulo Forattini

    1994-10-01

    Full Text Available Studies on breeding Anopheles albitarsis and association with rice growth in irrigated paddy fields were carried out during the rice cultivation cycle from December 1993 to March 1994. This period corresponded to the length of time of permanent paddy flooding. Breeding occurred in the early stage up until five weeks after transplantation when rice plant height was small. That inverse correlation may give potential direction to control measures.

  19. Assessment of Anopheles salivary antigens as individual exposure biomarkers to species-specific malaria vector bites

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    Ali Zakia M I

    2012-12-01

    Full Text Available Abstract Background Malaria transmission occurs during the blood feeding of infected anopheline mosquitoes concomitant with a saliva injection into the vertebrate host. In sub-Saharan Africa, most malaria transmission is due to Anopheles funestus s.s and to Anopheles gambiae s.l. (mainly Anopheles gambiae s.s. and Anopheles arabiensis. Several studies have demonstrated that the immune response against salivary antigens could be used to evaluate individual exposure to mosquito bites. The aim of this study was to assess the use of secreted salivary proteins as specific biomarkers of exposure to An. gambiae and/or An. funestus bites. Methods For this purpose, salivary gland proteins 6 (SG6 and 5′nucleotidases (5′nuc from An. gambiae (gSG6 and g-5′nuc and An. funestus (fSG6 and f-5′nuc were selected and produced in recombinant form. The specificity of the IgG response against these salivary proteins was tested using an ELISA with sera from individuals living in three Senegalese villages (NDiop, n = 50; Dielmo, n = 38; and Diama, n = 46 that had been exposed to distinct densities and proportions of the Anopheles species. Individuals who had not been exposed to these tropical mosquitoes were used as controls (Marseille, n = 45. Results The IgG responses against SG6 recombinant proteins from these two Anopheles species and against g-5′nucleotidase from An. gambiae, were significantly higher in Senegalese individuals compared with controls who were not exposed to specific Anopheles species. Conversely, an association was observed between the level of An. funestus exposure and the serological immune response levels against the f-5′nucleotidase protein. Conclusion This study revealed an Anopheles salivary antigenic protein that could be considered to be a promising antigenic marker to distinguish malaria vector exposure at the species level. The epidemiological interest of such species-specific antigenic markers is discussed.

  20. "Repellent Effect of Extracts and Essential Oils of Citrus limon (Rutaceae and Melissa officinalis (Labiatae Against Main Malaria Vector, Anopheles stephensi (Diptera: Culicidae"

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    "MA Oshaghi

    2003-10-01

    Full Text Available Repellet effect of extracts and essential oils of Citrus limon (L. Burm.F., (lemon and Melissa officinalis, (balm were evaluated against Anopheles stephensi in laboratory on animal and human and compared with synthetic repellent, N,Ndiethyl- 3-methylbenzamide (Deet as a standard. Results of statistical analysis revealed significant differences between oils and extracts (P< 0.05 against the tested species, thus oils were more effective than extracts. There was no significant difference between Deet and lemon oil, whereas the difference between lemon and melissa oils was significant. Relative efficacy of lemon oil to Deet was 0.88 whereas it was 0.71 for melissa oil. The results were found marginally superior in repellency for animals than human. Due to advantages of botanic compounds to synthetic compounds we recommend lemon essential oil as an effective alternative to Deet with potential as a means of personal protection against mosquito vectors of disease.

  1. Comparison of two commercial formulations of Bacillus thuringiensis var. israelensis for the control of Anopheles aquasalis (Diptera: Culicidae at three salt concentrations

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    Frances R Osborn

    2007-02-01

    Full Text Available Anopheles aquasalis larvae are salt water tolerant, preferring concentrations between 10 and 20 parts per thousand (ppt. The larvicidal efficacy of two formulations of Bacillus thuringiensis var. israelensis (Vectobac-12AS® and Bactivec®, was investigated against An. aquasalis at salinities of 0, 10, and 20 ppt. A probit analysis was used to calculate the lethal concentrations (LC50 and LC95 for each product at each salinity. The LC50 and LC95 were higher for Bactivec® than Vectobac-12AS®, and for Bactivec®, the LC50 and LC95 increased with salinity. Vectobac-12AS® should thus be preferred to Bactivec® for An. aquasalis control, especially in saline breeding habitats.

  2. Effect of anti-mosquito hemolymph antibodies on fecundity and on the infectivity of malarial parasite Plasmodium vivax to Anopheles stephensi (Diptera:Insecta).

    Science.gov (United States)

    Gulia, Monika; Suneja, Amita; Gakhar, Surendra K

    2002-06-01

    Rabbit antibodies to hemolymph antigens (102.5, 101, 100, 96, 88, 80, 64, 55, 43, 29, and 23 kDa) of Anopheles stephensi reduced fecundity as well as viability in An. stephensi. However, ingestion of these antibodies was not associated with a marked effect on the engorgement of mosquitoes but egg laying was significantly delayed. Antisera raised against hemolymph proteins were also used to identify cross reactive antigens/epitopes present in other tissues by Western blotting, as well as by in vivo ELISA. In addition, a significant reduction in oocyst development was also observed in An. stephensi mosquitoes that ingested anti-hemolymph antibodies along with Plasmodium vivax. The results confirmed the feasibility of targeting mosquito antigens as a novel anti-mosquito strategy, as well as confirmed the usefulness of such antigens for the development of a transmission-blocking vaccine. PMID:12195047

  3. Dynamics of immature stages of Anopheles arabiensis and other mosquito species (Diptera: Culicidae) in relation to rice cropping in a rice agro-ecosystem in Kenya.

    Science.gov (United States)

    Mwangangi, Joseph; Shililu, Josephat; Muturi, Ephantus; Gu, Weidong; Mbogo, Charles; Kabiru, Ephantus; Jacob, Benjamin; Githure, John; Novak, Robert

    2006-12-01

    We determined changes in species composition and densities of immature stages of Anopheles arabiensis mosquitoes in relation to rice growth cycle in order to generate data for developing larval control strategies in rice ecosystems. Experimental rice paddies (6.3m x 3.15m) exposed to natural colonization of mosquitoes were sampled weekly for two rice growing cycles between February 2004 and March 2005. Overall, 21,325 Anopheles larvae were collected, of which 91.9% were 1st and 2nd instars and 8.1% were 3rd and 4th instars. An. arabiensis was the predominant species (84.1%) with other species, An. pharoensis (13.5%), An. funestus (2.1%), An. coustani (0.3%), and An. maculipalpis (0.1%) accounting for only a small proportion of the anophelines collected. Culex quinquefasciatus (65.7%) was the predominant species among the non-anopheline species. Others species collected included: C. annulioris (9.9%), C. poicilipes (7.3%), C. tigripes (7.2%), C. duttoni (0.6%), Aedes aegypti (5.3%), Ae. cumminsii (3.5%), and Ae. vittatus (0.7%). The densities of the major anopheline species were closely related to rice stage and condition of the rice field. An. arabiensis, the predominant species, was most abundant over a three-week period after transplanting. Low densities of larvae were collected during the late vegetative, reproductive, and ripening phases of rice. An increase in larval density ten days post-transplanting was found to correlate with the application of fertilizer (sulphate of ammonia). Culicine and aedine species densities were significantly higher during the post-harvesting period. Our results suggest that the transplanting stage is favorable for the growth of immature stages of An. arabiensis and provides a narrow window for targeted larval intervention in rice. PMID:17249341

  4. Anopheles salivary gland proteomes from major malaria vectors

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    Fontaine Albin

    2012-11-01

    Full Text Available Abstract Background Antibody responses against Anopheles salivary proteins can indicate individual exposure to bites of malaria vectors. The extent to which these salivary proteins are species-specific is not entirely resolved. Thus, a better knowledge of the diversity among salivary protein repertoires from various malaria vector species is necessary to select relevant genus-, subgenus- and/or species-specific salivary antigens. Such antigens could be used for quantitative (mosquito density and qualitative (mosquito species immunological evaluation of malaria vectors/host contact. In this study, salivary gland protein repertoires (sialomes from several Anopheles species were compared using in silico analysis and proteomics. The antigenic diversity of salivary gland proteins among different Anopheles species was also examined. Results In silico analysis of secreted salivary gland protein sequences retrieved from an NCBInr database of six Anopheles species belonging to the Cellia subgenus (An. gambiae, An. arabiensis, An. stephensi and An. funestus and Nyssorhynchus subgenus (An. albimanus and An. darlingi displayed a higher degree of similarity compared to salivary proteins from closely related Anopheles species. Additionally, computational hierarchical clustering allowed identification of genus-, subgenus- and species-specific salivary proteins. Proteomic and immunoblot analyses performed on salivary gland extracts from four Anopheles species (An. gambiae, An. arabiensis, An. stephensi and An. albimanus indicated that heterogeneity of the salivary proteome and antigenic proteins was lower among closely related anopheline species and increased with phylogenetic distance. Conclusion This is the first report on the diversity of the salivary protein repertoire among species from the Anopheles genus at the protein level. This work demonstrates that a molecular diversity is exhibited among salivary proteins from closely related species despite their

  5. Revisión histórica de la distribución de Anopheles (Nyssorhynchus darlingi (Diptera: Culicidae en la amazonía peruana

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    Roberto Fernández

    2014-06-01

    Full Text Available Anopheles (Nyssorhynchus darlingi ha sido reportado desde 1931 en áreas fronterizas del departamento de Loreto, principalmente a lo largo de las fronteras con Brasil y Colombia. En 1994, durante un brote de malaria, An. darlingi fue encontrado en localidades vecinas a Iquitos. En la actualidad, su distribución se ha expandido considerablemente en Loreto. Para los objetivos de este trabajo se realizó una revisión de las referencias bibliográficas disponibles para obtener toda la información posible sobre la distribución de los mosquitos, en particular de los anofelinos en la región Amazónica del país, con énfasis especial en An. darlingi. También se realizaron colectas entomológicas en los departamentos de Madre de Dios y Ucayali con la finalidad de conocer y verificar la distribución de An. darlingi. En la actualidad, la distribución de la especie está circunscrita a localidades al sureste del Perú en localidades fronterizas con Bolivia, en una localidad del río Abujao del departamento de Ucayali, y ampliamente en la región noreste de la cuenca amazónica del departamento de Loreto

  6. Revisión histórica de la distribución de Anopheles (Nyssorhynchus darlingi (Diptera: Culicidae en la amazonía peruana

    Directory of Open Access Journals (Sweden)

    Roberto Fernández

    2014-04-01

    Full Text Available Anopheles (Nyssorhynchus darlingi ha sido reportado desde 1931 en áreas fronterizas del departamento de Loreto, principalmente a lo largo de las fronteras con Brasil y Colombia. En 1994, durante un brote de malaria, An. darlingi fue encontrado en localidades vecinas a Iquitos. En la actualidad, su distribución se ha expandido considerablemente en Loreto. Para los objetivos de este trabajo se realizó una revisión de las referencias bibliográficas disponibles para obtener toda la información posible sobre la distribución de los mosquitos, en particular de los anofelinos en la región Amazónica del país, con énfasis especial en An. darlingi. También se realizaron colectas entomológicas en los departamentos de Madre de Dios y Ucayali con la finalidad de conocer y verificar la distribución de An. darlingi. En la actualidad, la distribución de la especie está circunscrita a localidades al sureste del Perú en localidades fronterizas con Bolivia, en una localidad del río Abujao del departamento de Ucayali, y ampliamente en la región noreste de la cuenca amazónica del departamento de Loreto

  7. Mosquitocidal activity of indigenenous plants of Western Ghats, Achras sapota Linn. (Sapotaceae) and Cassia auriculata L. (Fabaceae) against a common malarial vector, Anopheles stephensi Liston (Culicidae:Diptera)

    Institute of Scientific and Technical Information of China (English)

    Kaliyamoorthy Krishnappa; Kuppusamy Elumalai

    2014-01-01

    Objective: To evaluate the mosquito larvicidal, ovicidal, pupicidal and repellent activities of hexane, diethyl ether, dichloromethane, acetone and methanol extracts of Indian medicinal plants, Achras sapota (A. sapota) and Cassia auriculata (C. auriculata) at different concentrations against Anopheles stephensi (An. stephensi), a malarial vector. Methods: Twenty five early third instar larvae of An. stephensi were exposed to various concentrations (30-210 mg/L) of plants extracts and were assayed in the laboratory by using the protocol of WHO 2005;then after 24 h LC50 values of the A. sapota and C. auriculata leaf extract was determined by probit analysis. The ovicidal activity was tested with the extracts ranging from 50-350 mg/L. The pupicidal activity was recorded after 24 h of exposure to the extract. The repellent efficacy was determined against mosquito species at two different concentrations 1.5 and 3.0 mg/cm2 under laboratory conditions. Results:Among the five different extracts tested against the An. stephensi, methanol extract of A. sapota proved to be an more effective solvent extract in almost all the parameters studied than C. auriculata. Conclusions:It is inferred that the leaf extract of A. sapota and C. auriculata could be used in vector control programme.

  8. Production of a Transgenic Mosquito Expressing Circumsporozoite Protein, a Malarial Protein, in the Salivary Gland of Anopheles stephensi (Diptera:Culicidae

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    Matsuoka,Hiroyuki

    2010-08-01

    Full Text Available We are producing a transgenic mosquito, a flying syringe, to deliver a vaccine protein to human beings via the saliva the mosquito deposits in the skin while biting. The mosquito produces a vaccine protein in the salivary gland (SG and deposits the protein into the host's skin when it takes the host's blood. We chose circumsporozoite protein (CSP, currently the most promising malaria vaccine candidate, to be expressed in the SG of Anopheles stephensi. To transform the mosquitoes, plasmid containing the CSP gene under the promoter of female SG-specific gene, as well as the green fluorescent protein (GFP gene under the promoter of 3xP3 as a selection marker in the eyes, was injected into more than 400 eggs. As a result, five strains of GFP-expressing mosquitoes were established, and successful CSP expression in the SG was confirmed in one strain. The estimated amount of CSP in the SG of the strain was 40ng per mosquito. We allowed the CSP-expressing mosquitoes to feed on mice to induce the production of anti-CSP antibody. However, the mice did not develop anti-CSP antibody even after transgenic mosquitoes had bitten them several times. We consider that CSP in the SG was not secreted properly into the saliva. Further techniques and trials are required in order to realize vaccine-delivering mosquitoes.

  9. Insecticidal potential of Ocimum canum plant extracts against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus larval and adult mosquitoes (Diptera: Culicidae).

    Science.gov (United States)

    Murugan, Jimmantiyur Madhappan; Ramkumar, Govindaraju; Shivakumar, Muthugoundar Subramanian

    2016-01-01

    Mosquitoes have developed resistance to various synthetic insecticides, making their control increasingly difficult. Insecticides of botanical origin may serve as suitable natural control. This study evaluates the toxic potential of Ocimum canum (Sims) leaf extract and powder against Anopheles stephensi (Liston), Aedes aegypti (Lin) and Culex quinquefasciatus (Say) larval and adult mosquitoes. Larval mortality was observed after 24 h recovery period and adult smoke toxicity observed for 40 min duration at 10 min interval. Methanol extract of O. canum showed highest larval mortality against the larvae of C. quinquefasciatus LC50 = 28.3225, LC90 = 44.1150; Ae. aegypti LC50 = 43.327, LC90 = 61.249; and An. stephensi LC50 = 30.2001, LC90 = 48.2866 ppm. The smoke toxicities were 93% mortality in C. quinquefasciatus, 74% in Ae. aegypti and 79% in An. stephensi adults, respectively, whereas 100% mortality was recorded in the commercial mosquito control. Our results suggest that O. canum leaf extract and powder are natural insecticide, and ideal eco friendly approach for mosquito control.

  10. Field investigation on the repellent activity of some aromatic plants by traditional means against Anopheles arabiensis and An. pharoensis (Diptera: Culicidae) around Koka, central Ethiopia.

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    Dugassa, Sisay; Medhin, Girmay; Balkew, Meshesha; Seyoum, Aklilu; Gebre-Michael, Teshome

    2009-10-01

    A study was undertaken to evaluate the impact of traditional application methods of mosquito repellent plants in the reduction of the human-vector contact of malaria vectors in central Ethiopia. The plants (Corymbia citriodora, Eucalyptus camaldulensis, Ocimum suave and Ocimum basilicum) were tested by thermal expulsion and direct burning on traditional stoves in the field against two important malaria vectors in Ethiopia (Anopheles arabiensis and An. pharoensis). A Latin-square design was applied for randomly assigning the treatment plants and control to experimental houses over different nights. The percentage repellency of each candidate plant by both application methods was estimated from the catches of mosquitoes in the treatment and control houses. On direct burning of the plants, O. basilicum showed the highest percentage repellency (73.11%, Pcitriodora on the other hand gave the highest repellency (72.87%, Pcitriodora exhibited the highest repellency (78.69%, Pcitriodora gave the highest repellency (72.9%, P65%) against the house-entry and biting of two important malaria vectors in Ethiopia, and thus have a potential to be used at least as supplements to other control methods. However, feasibility and actual impact on disease transmission need to be known on these and other potentially useful plants. PMID:19539591

  11. First Record of Anopheles darlingi Root (Diptera, Culicidae in the Volta Grande Environmental Reserve, Conceição das Alagoas Municipality, Minas Gerais, Brazil

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    Rafael Martins

    2015-04-01

    Resumo. Três espécimes de Anopheles darlingi Root foram encontrados na reserva ambiental Volta Grande, na área de influência da barragem do Rio Grande construída pela usina hidrelétrica de Volta Grande, em Conceição das Alagoas, Minas Gerais, Brasil. O mosquito An. darlingi é um dos principais vetores da malária humana no Brasil, devido à sua preferência alimentar por sangue humano, um fator que é acentuado pelo comportamento endofílico da espécie. O presente relato poderá ser útil à vigilância entomológica local para monitoramento dos impactos gerados pela formação do reservatório hidrelétrico, uma vez que a presença deste vetor neste tipo de ambiente indica um risco potencial de transmissão da malária.

  12. The Influence of Insecticide Resistance, Age, Sex, and Blood Feeding Frequency on Thermal Tolerance of Wild and Laboratory Phenotypes of Anopheles funestus (Diptera: Culicidae).

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    Lyons, C L; Oliver, S V; Hunt, R H; Coetzee, M

    2016-03-01

    Resistance to insecticides is a global phenomenon and is increasing at an unprecedented rate. How resistant and susceptible strains of malaria vectors might differ in terms of life history and basic biology is often overlooked, despite the potential importance of such information in light of changing climates. Here, we investigated the upper thermal limits (ULT50) of wild and laboratory strains of Anopheles funestus Giles mosquitoes, including resistance status, sex, age, and blood feeding status as potential factors influencing ULT50. No significant differences in ULT50 were observed between strains displaying different resistance patterns, nor was there a significant difference between wild and laboratory strains. In some instances, strains showed a senescence response, displaying decreased ULT50 with an increase in age, and differences between males and females (females displaying higher ULT50 than males). Blood feeding did not seem to influence ULT50 in any way. For An. funestus, it seems evident that there is no cost to resistance despite what is displayed in other anopheline species. This could have significant impacts for vector control, with resistant populations of An. funestus performing just as well, if not better, than susceptible strains, especially under changing environmental conditions such as those expected to occur with climate change. PMID:26718714

  13. Survival of anopheline eggs and their susceptibility to infection with Metarhizium anisopliae and Beauveria bassiana under laboratory conditions

    NARCIS (Netherlands)

    Luz, C.; Mnyone, L.L.; Russell, T.L.

    2011-01-01

    The viability of Anopheles gambiae sensu stricto and Anopheles arabiensis (Diptera: Culicidae) eggs over time and the ovicidal activity of Beauveria bassiana (Ascomycota: Cordycipitaceae) and Metarhizium anisopliae (Ascomycota: Clavicipitaceae) were investigated. Eggs were incubated in soil or leaf

  14. Larvicidal,ovicidal and repellent activities of marine sponge Cliona celata(Grant) extracts against Anopheles stephensi Liston(Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    Appadurai; Daniel; Reegan; Arokia; Valan; Kinsalin; Michael; Gabriel; Paulraj; Savarimuthu; Ignacimuthu

    2015-01-01

    Obective:To evaluate the larvieidal.ovieidal and repellent properties of solvent extracts of marine sponge Cliona celala(C.celata)(Grant) against the malarial vector Anopheles stephensi(An.stephensi) Listen.Methods:Marine sponge C.celala was thoroughly washed with distilled water and shade dried for 48 h.Then the sponges were homogenized and extracted sequentially with hexane,ethyl acetate and methanol,Lanvicidal and ovicidal activities were tested at four different concentrations viz.,62.5.125.0.250.0 and 500.0 ppm.for repellent study extracts were taken in three different concentrations viz..5.0.2.5.1.0 mg/cm at.Results:Among the three solvent extracts of C.celata,methanol extract showed thc highest lanvieidal activity at 500 ppm against the fourth inslar larvae of An.stephensi.The LC50and LC90 values of C.celata methanol extract were recorded as 80.61 and 220.81 ppm against An.stephensi larvae respectively.High ovieidal activity of 91.2%was recorded al 500 ppm concentration of methanol extract.The haxane extract was found to be the most effective protectant against the adult female mosquitoes of An.stephensi.The mean protection time recorded in hexane extract was up to 245 min at 5 mg/cm2 dosage against An.Stephensi adults.Conclusions:The screening results suggest that the hexane and methanol extracts of C.celata arc promising in mosquito control.Considering these bioaclivitics.C.celata could the probed further to obtain some novel pesticidal molecules.

  15. Efficacy of larvicidal and pupicidal activity of Catharanthus roseus aqueous and solvent extracts against Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    Selladurai Subarani; Selvi Sabhanayakam; Chinnaperumal Kamaraj; Gandhi Elango; Mohamed Abdul Kadir

    2013-01-01

    Objective:To investigate the larvicidal and pupicidal activities of aqueous, ethyl acetate and methanol extracts of Catharanthus roseus (C. roseus) against malaria and filariasis vectors. Methods:The larvicidal and pupicidal activities of C. roseus leaf extracts were tested against the fourth instar larvae and pupae of Anopheles stephensi (An. stephensi) and Culex quinquefasciatus (Cx. quinquefasciatus). The mortality was observed after 24 and 48 h post the treatment. The data were subjected to probit analysis to determine the lethal concentrations (LC50 and LC90) at which 50%and 90%of the treated larvae or pupae of the tested species were killed. Results:The larval and pupal mortality were observed after 24 and 48 h of exposure of aqueous, ethyl acetate and methanol extracts of C. roseus; no mortality was observed in the control group. The LC50 values against the fourth-instar larvae of An. stephensi were 68.62 and 72.04 mg/mL for the aqueous extract, 82.47 mg/mL for the ethyl acetate extract, and 78.80 and 86.64 mg/mL for the methanol extract, while the aqueous, ethyl acetate and methanol extracts had LC50 values of 85.21, 76.84 and 94.20 mg/mL against the fourth-instar larvae of Cx. quinquefasciatus. The aqueous, ethyl acetate and methanol extracts had LC50 values of 118.08, 182.47 and 143.80 mg/mL against the pupae of An. stephensi and 146.20, 226.84 and 156.62 mg/mL against the pupae of Cx. quinquefasciatus, respectively. Conclusions: The aqueous and methanol extracts of C. roseus leaves had an excellent potential to control the malarial vector An. stephensi and filariasis vector Cx. quinquefasciatus.

  16. Larvicidal, ovicidal and pupicidal activities ofGliricidia sepium (Jacq.) (Leguminosae) against the malarial vector,Anopheles stephensiListon (Culicidae:Diptera)

    Institute of Scientific and Technical Information of China (English)

    Kaliyamoorthy Krishnappa; Shanmugam Dhanasekaran; Kuppusamy Elumalai

    2012-01-01

    Objective:To investigate the potentiality of mosquitocidal activity ofGliricidia sepium(G. sepium) (Jacq.)(Leguminosae).Methods:Twenty five early third instar larvae ofAnopheles stephensi (An. stephensi) were exposed to various concentrations(50-250 ppm) and the24 hLC50 values of theG. sepium extract was determined by probit analysis.The ovicidal activity was determined againstAn. stephensi to various concentrations ranging from25-100 ppm under laboratory conditions.The eggs hatchability was assessed48 h post treatment.The pupicidal activity was determined againstAn. stephensi to various concentrations ranging from25-100 ppm.Mortality of each pupa was recorded after24 h of exposure to the extract.Results:Results pertaining to the experiment clearly revealed that ethanol extract showed significant larvicidal, ovicidal and pupicidal activity against theAn. stephensi.Larvicidal activityof ethanol extracts ofG. sepium showed maximum mortality in250 ppm concentration(96.0±2.4)%.Furthermore, theLC50 was found to be121.79 and theLC90value was recorded to be231.98 ppm.Ovicidal activity of ethanol extract was assessed by assessing the egg hatchability.Highest concentration of both solvent extracts exhibited100% ovicidal activity.Similarly, pupae exposed to different concentrations of ethanol extract were found dead with58.10% adult emergence when it was treated with25 ppm concentration.Similarly,18.36(n=30;61.20%);21.28(70.93) and27.33(91.10) pupal mortality was recorded from the experimental pupae treated with50,75 and100 ppm concentration of extracts. Three fractions have been tested for their larvicidal activity of which theFraction3 showed the LC50 andLC90 values of23.23 and40.39 ppm.With regard to the ovicidal effect fraction3 showed highest ovicidal activities than the other two fractions.Furthermore, there were no hatchability was recorded above50 ppm(100% egg mortality) in the experimental group.Statistically significant pupicidal activity was recorded from75 ppm

  17. Larvicidal and Biting Deterrent Activity of Essential Oils of Curcuma longa, Ar-turmerone, and Curcuminoids Against Aedes aegypti and Anopheles quadrimaculatus (Culicidae: Diptera).

    Science.gov (United States)

    Ali, Abbas; Wang, Yan-Hong; Khan, Ikhlas A

    2015-09-01

    Essential oils and extract of Curcuma longa, ar-turmerone, and curcuminoids were evaluated for their larvicidal and deterrent activity against mosquitoes. Ar-turmerone and curcuminoids constituted 36.9, 24.9 and 50.6% of rhizome oil, leaf oil, and rhizome extract, respectively. Ar-turmerone was the major compound of the rhizome oil (36.9%) and leaf oil (24.9%). The ethanolic extract had 15.4% ar-turmerone with 6.6% bisdesmethoxycurcumin, 6.1% desmethoxycurcumin, and 22.6% curcumin. In in vitro studies, essential oils of the leaf (biting deterrence index [BDI] = 0.98), rhizome (BDI = 0.98), and rhizome ethanolic extract (BDI = 0.96) at 10 µg/cm(2) showed biting deterrent activity similar to DEET at 25 nmol/cm(2) against Aedes aegypti L. Among the pure compounds, ar-turmerone (BDI = 1.15) showed the biting deterrent activity higher than DEET at 25 nmol/cm(2) whereas the activity of other compounds was lower than DEET. In Anopheles quadrimaculatus Say, only ar-turmerone showed deterrent activity similar to DEET. In dose-response bioassay, ar-turmerone showed significantly higher biting deterrence than DEET at all the dosages. Ar-turmerone, at 15 nmol/cm(2), showed activity similar to DEET at 25 nmol/cm(2) and activity at 5 nmol/cm(2) was similar to DEET at 20 and 15 nmol/cm(2). Leaf essential oil with LC(50) values of 1.8 and 8.9 ppm against larvae of An. quadrimaculatus and Ae. aegypti, respectively, showed highest toxicity followed by rhizome oil and ethanolic extract. Among the pure compounds, ar-turmerone with LC(50) values of 2.8 and 2.5 ppm against larvae of An. quadrimaculatus and Ae. aegypti, respectively, was most toxic followed by bisdesmethoxycurcumin, curcumin, and desmethoxycurcumin.

  18. The abundance and host-seeking behavior of culicine species (Diptera: Culicidae) and Anopheles sinensis in Yongcheng city, people's Republic of China

    Science.gov (United States)

    2011-01-01

    Background The knowledge of mosquito species diversity and the level of anthropophily exhibited by each species in a region are of great importance to the integrated vector control. Culicine species are the primary vectors of Japanese encephalitis (JE) virus and filariasis in China. Anopheles sinensis plays a major role in the maintenance of Plasmodium vivax malaria transmission in China. The goal of this study was to compare the abundance and host-seeking behavior of culicine species and An. sinensis in Yongcheng city, a representative region of P. vivax malaria. Specifically, we wished to determine the relative attractiveness of different animal baits versus human bait to culicine species and An. sinensis. Results Culex tritaeniorhynchus was the most prevalent mosquito species and An. sinensis was the sole potential vector of P. vivax malaria in Yongcheng city. There were significant differences (P < 0.01) in the abundance of both An. sinensis and Cx. tritaeniorhynchus collected in distinct baited traps. The relative attractiveness of animal versus human bait was similar towards both An. sinensis and Cx. tritaeniorhynchus. The ranking derived from the mean number of mosquitoes per bait indicated that pigs, goats and calves frequently attracted more mosquitoes than the other hosts tested (dogs, humans, and chickens). These trends were similar across all capture nights at three distinct villages. The human blood index (HBI) of female An. sinensis was 2.94% when computed with mixed meals while 3.70% computed with only the single meal. 19:00~21:00 was the primary peak of host-seeking female An. sinensis while 4:00~5:00 was the smaller peak at night. There was significant correlation between the density of female An. sinensis and the average relative humidity (P < 0.05) in Wangshanzhuang village. Conclusions Pigs, goats and calves were more attractive to An. sinensis and Cx. tritaeniorhynchus than dogs, humans, and chickens. Female An. sinensis host-seeking activity

  19. The abundance and host-seeking behavior of culicine species (Diptera: Culicidae and Anopheles sinensis in Yongcheng city, people's Republic of China

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    Liu Xiao-Bo

    2011-11-01

    Full Text Available Abstract Background The knowledge of mosquito species diversity and the level of anthropophily exhibited by each species in a region are of great importance to the integrated vector control. Culicine species are the primary vectors of Japanese encephalitis (JE virus and filariasis in China. Anopheles sinensis plays a major role in the maintenance of Plasmodium vivax malaria transmission in China. The goal of this study was to compare the abundance and host-seeking behavior of culicine species and An. sinensis in Yongcheng city, a representative region of P. vivax malaria. Specifically, we wished to determine the relative attractiveness of different animal baits versus human bait to culicine species and An. sinensis. Results Culex tritaeniorhynchus was the most prevalent mosquito species and An. sinensis was the sole potential vector of P. vivax malaria in Yongcheng city. There were significant differences (P An. sinensis and Cx. tritaeniorhynchus collected in distinct baited traps. The relative attractiveness of animal versus human bait was similar towards both An. sinensis and Cx. tritaeniorhynchus. The ranking derived from the mean number of mosquitoes per bait indicated that pigs, goats and calves frequently attracted more mosquitoes than the other hosts tested (dogs, humans, and chickens. These trends were similar across all capture nights at three distinct villages. The human blood index (HBI of female An. sinensis was 2.94% when computed with mixed meals while 3.70% computed with only the single meal. 19:00~21:00 was the primary peak of host-seeking female An. sinensis while 4:00~5:00 was the smaller peak at night. There was significant correlation between the density of female An. sinensis and the average relative humidity (P Conclusions Pigs, goats and calves were more attractive to An. sinensis and Cx. tritaeniorhynchus than dogs, humans, and chickens. Female An. sinensis host-seeking activity mainly occurred from 19:00 to 21:00. Thus

  20. Low-cost and eco-friendly green synthesis of silver nanoparticles using Feronia elephantum (Rutaceae) against Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    Veerakumar, Kaliyan; Govindarajan, Marimuthu; Rajeswary, Mohan; Muthukumaran, Udaiyan

    2014-05-01

    Mosquitoes transmit serious human diseases, causing millions of deaths every year. The use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of synthesized natural products for vector control have been a priority in this area. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Feronia elephantum plant leaf extract against late third-instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus was determined. The range of concentrations of synthesized AgNPs (5, 10, 15, 20, and 25 μg mL(-1)) and aqueous leaf extract (25, 50, 75, 100, and 125 μg mL(-1)) were tested against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of F. elephantum for all three important vector mosquitoes. The synthesized AgNPs from F. elephantum were highly toxic than crude leaf aqueous extract to three important vector mosquito species. The results were recorded from UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy analysis (EDX). Synthesized AgNPs against the vector mosquitoes A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC50 and LC90 values: A. stephensi had LC50 and LC90 values of 11.56 and 20.56 μg mL(-1); A. aegypti had LC50 and LC90 values of 13.13 and 23.12 μg mL(-1); and C. quinquefasciatus had LC50 and LC90 values of 14.19 and 24.30 μg mL(-1). No mortality was observed in the control. These results suggest that the green synthesis of silver nanoparticles using F. elephantum has the potential to be used as an ideal eco-friendly approach for the control of A. stephensi, A. aegypti, and C

  1. Mosquito larvicidal potential of silver nanoparticles synthesized using Chomelia asiatica (Rubiaceae) against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae).

    Science.gov (United States)

    Muthukumaran, Udaiyan; Govindarajan, Marimuthu; Rajeswary, Mohan

    2015-03-01

    Mosquitoes transmit serious human diseases, causing millions of deaths every year. Mosquito control is to enhance the health and quality of life of county residents and visitors through the reduction of mosquito populations. Mosquito control is a serious concern in developing countries like India due to the lack of general awareness, development of resistance, and socioeconomic reasons. Today, nanotechnology is a promising research domain which has a wide ranging application in vector control programs. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, larvicidal activity of aqueous leaf extract and silver nanoparticles (AgNPs) synthesized using C. asiatica plant leaves against late third instar larvae of Anopheles stephensi, Aedes aegypti, and Cx. quinquefasciatus. The range of varying concentrations of synthesized AgNPs (8, 16, 24, 32, and 40 μg/mL) and aqueous leaf extract (40, 80, 120, 160, and 200 μg/mL) were tested against the larvae of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus. The synthesized AgNPs from C. asiatica were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV-Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy analysis (EDX). Considerable mortality was evident after the treatment of C. asiatica for all three important vector mosquitoes. The LC50 and LC90 values of C. asiatica aqueous leaf extract appeared to be effective against An. stephensi (LC50, 90.17 μg/mL; LC90, 165.18 μg/mL) followed by Ae. aegypti (LC50, 96.59 μg/mL; LC90, 173.83 μg/mL) and Cx. quinquefasciatus (LC50, 103.08 μg/mL; LC90, 183.16 μg/mL). Synthesized AgNPs against the vector mosquitoes of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus had the following LC50 and LC90

  2. Green synthesis of silver nanoparticles using Sida acuta (Malvaceae) leaf extract against Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    Veerakumar, Kaliyan; Govindarajan, Marimuthu; Rajeswary, Mohan

    2013-12-01

    Mosquitoes act as a vector for most of the life-threatening diseases like malaria, yellow fever, dengue fever, chikungunya fever, filariasis, encephalitis, West Nile Virus infection, etc. Under the Integrated Mosquito Management, emphasis was given on the application of alternative strategies in mosquito control. The continuous application of synthetic insecticides causes development of resistance in vector species, biological magnification of toxic substances through the food chain, and adverse effects on environmental quality and nontarget organisms including human health. Application of active toxic agents from plant extracts as an alternative mosquito control strategy was available from ancient times. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Sida acuta plant leaf extract against late third instar larvae of Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti was determined. Range of concentrations of synthesized AgNPs (10, 20, 30, 40, and 50 μg/mL) and aqueous leaf extract (50, 100, 150, 200, and 250 μg/mL) were tested against the larvae of C. quinquefasciatus, A. stephensi and A. aegypti. The synthesized AgNPs from S. acuta leaf were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV-Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of S. acuta for all three important vector mosquitoes. The LC50 and LC90 values of S. acuta aqueous leaf extract appeared to be most effective

  3. Studies on the impact of biosynthesized silver nanoparticles (AgNPs) in relation to malaria and filariasis vector control against Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae).

    Science.gov (United States)

    Subarani, Selladurai; Sabhanayakam, Selvi; Kamaraj, Chinnaperumal

    2013-02-01

    Biosynthesized nanoparticles have been achieved using environmentally acceptable plant extract and eco-friendly reducing and capping agents. The present study was based on assessments of the larvicidal activities to determine the efficacies of synthesized silver nanoparticles (AgNPs) using aqueous leaf extract of Vinca rosea (L.) (Apocynaceae) against the larvae of malaria vector Anopheles stephensi Liston and filariasis vector Culex quinquefasciatus Say (Diptera: Culicidae). Larvae were exposed to varying concentrations of aqueous extract of V. rosea and synthesized AgNPs for 24, 48, and 72 h. AgNPs were rapidly synthesized using the leaf extract of V. rosea, and the formation of nanoparticles was observed within 15 min. The results recorded from UV-Vis spectrum, Fourier transform infrared (FTIR), X-ray diffraction (XRD), scanning electron microscopy (SEM), and transmission electron microscopy (TEM) support the biosynthesis and characterization of AgNPs. The formation of the AgNPs synthesized from the XRD spectrum compared with the Bragg reflections at 2θ = 29.36, 38.26, 44.51, 63.54, and 77.13° which can be indexed to the (121), (111), (200), (220), and (311) orientations, respectively, confirmed the presence of AgNPs. The FTIR spectra of AgNPs exhibited prominent peaks at the spectra showed sharp and strong absorption band at 3,406.71 to 3,431.90 cm(-1) double in case of NH(2) group of a primary amine (N-H stretch). The presence of the sharp peak at 2,926.54 to 2,925.80 cm(-1) very broad often looks like distorted baseline (O-H carboxylic acids). The band 1,633.26 to 1,625.81 cm(-1) was assigned to C = C alkenes, aromatic ring stretching vibration, respectively. SEM analysis of the synthesized AgNPs clearly showed the clustered and irregular shapes, mostly aggregated and having the size of 120 nm. TEM reveals spherical shape of synthesized AgNPs. Particle size analysis revealed that the size of particles ranges from 25 to 47 nm with average size of 34.61 nm

  4. Determinação das fontes alimentares de Anopheles aquasalis (Diptera: Culicidae no Estado do Rio de Janeiro, Brasil, pelo teste de precipitina Blood-meal sources of Anopheles aquasalis (Diptera: Culicidae in a South-eastern State of Brazil

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    Carmen Flores-Mendoza

    1996-04-01

    Full Text Available Anopheles aquasalis é um mosquito ora encarado como antropofílico, ora como zoofílico ou eclético. Realizou-se estudo em Guapimirim, Estado do Rio de Janeiro, de maio a novembro de 1992, com o intuito de se verificar a fonte alimentar preferida desse anofelino através de teste imunológico de precipitina. De 1.366 fêmeas capturadas em abrigos naturais, 725 estavam ingurgitadas. O conteúdo digestivo de apenas 473 delas reagiu no teste de precipitina, sendo que em 75,3% dos casos foi identificada apenas uma fonte alimentar. Mais da metade dessas fêmeas havia se alimentado em boi (52,2%, enquanto poucas tinham sugado homem (1,1%. Por outro lado, 24,7% dos espécimes haviam se alimentado em mais de uma fonte sangüínea, principalmente boi e cavalo. Conclui-se que An. aquasalis é zoófilo nessa região do País, utilizando grande variedade de hospedeiros, porém preferindo se alimentar em animais de grande porte, especialmente o boi e cavalo.Anopheles aquasalis has shown local variations in blood-host preference in Brazil: it seems to be anthropophilic in the northeast and zoophilic or opportunistic in the Amazon and other regions. A study was carried out in Guapimirim county, State of Rio de Janeiro, from May to November 1992, for the purpose of identifying the blood meal source of this anopheline by capillary tube precipitin test. A total of 1,366 females were captured at natural resting-places, 725 of which were blood-fed. The gut content of 473 blood fed females reacted to the antisera used (human, cow, horse, pig, dog and chicken. The majority of the females ¾ 356 (75.3% ¾ had blood from only one source. A substantial bovine preference was observed ¾ 52.2% had fed on cows, 29.8% on horses, 10.7% on pigs, 4.5% on chickens and 1.7% on dogs, but only few had fed on man (1.1% and none on rats. On the other hand, 24.7% of the females had fed on more than one host, mainly on cows and horses. It was assumed that An. aquasalis is zoophilic

  5. Distribuição das espécies do gênero Anopheles (Diptera, Culicidae no Estado do Maranhão, Brasil Distribution of species from genus Anopheles (Diptera, Culicidae in the State of Maranhão, Brazil

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    José M. Macário Rebêlo

    2007-12-01

    Full Text Available Estudou-se a distribuição e diversidade de espécies de Anopheles em 123 municípios do Estado do Maranhão, Brasil. O método básico foi a captura de fêmeas dentro e nos arredores das habitações humanas, em intervalos compreendidos entre 18h e 6h, no período de janeiro de 1992 a dezembro de 2001. Foram capturados 84.467 exemplares distribuídos em 24 espécies, com o predomínio de A. triannulatus sensu lato (20.788, A. darlingi (19.083, A. nuneztovari (16.884, A. albitarsis s.l. (14.352, A. aquasalis (8.202 e A. evansae (2.885. As outras 18 espécies juntas representaram apenas 2,7%. As espécies encontradas no maior número de municípios foram: A. albitarsis s.l. (109 municípios, A. triannulatus s.l. (106, A. nuneztovari (93, A. darlingi (87 e A. evansae (64. A riqueza e a ampla distribuição das espécies de anofelinos no Maranhão concordam com a posição geográfica do estado, entre as macrorregiões que caracterizam o Brasil, resultando em uma fauna mista, com elementos representativos dessas regiões.We studied the distribution and diversity of Anopheles species in 123 counties (municipalities in the State of Maranhão, Brazil. The basic method consisted of capturing female specimens inside and around human dwellings between 6 PM and 6 AM from January 1992 to December 2001. A total of 84,467 specimens belonging to 24 species were captured, with a predominance of A. triannulatus sensu lato (20,788, A. darlingi (19,083, A. nuneztovari (16,884, A. albitarsis s.l. (14,352, A. aquasalis (8.202, and A. evansae (2,885. The other 18 species together accounted for only 2.7% of the total. The species found in the most counties were A. albitarsis s.l. (109 counties, A. triannulatus s.l. (106, A. nuneztovari (93, A. darlingi (87, and A. evansae (64. The richness and wide distribution of anopheline species in Maranhão agree with the State's geographic position among Brazil's macro-regions, resulting in a mixed fauna with representative

  6. Radiation-induced sterility for pupal and adult stages of the malaria moquito Anopheles arabiensis

    OpenAIRE

    Helinski, M.E.H.; Parker, A.G.; Knols, B.G.J.

    2006-01-01

    Background - In the context of the Sterile Insect Technique (SIT), radiation-induced sterility in the malaria mosquito Anopheles arabiensis Patton (Diptera: Culicidae) was studied. Male mosquitoes were exposed to gamma rays in the pupal or adult stage and dose-sterility curves were determined. Methods - Pupae were irradiated shortly before emergence (at 22-26 hrs of age), and adults

  7. Studies on mosquitoes (Diptera: Culicidae and anthropic environment: 1- Parity of blood seeking Anopheles (Kerteszia in South-Eastern Brazil Estudos sobre mosquitos (Diptera: Culicidae e ambiente antrópico: 1- Paridade de Anopheles (Kerteszia em atividade hematófaga, na região sudeste do Brasil

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    Oswaldo Paulo Forattini

    1993-02-01

    Full Text Available Populations of Anopheles (Kerteszia were sampled fortnightly over a one-year period (August 1991 to July 1992 at Ribeira Valley, S. Paulo State, Brazil. Indoor and outdoor collections were made on human bait at evening crepuscular period. The Polovodova technique for age grading was applied to 3,501 females of Anopheles cruzii and to 416 females of An. bellator. That sample represented 34.4% of the total number of mosquitoes collected. The most abundant species found was An. cruzii. However, An. bellator showed an endophagy that was almost three times greater than that of An. cruzii. The overall parous rate was 25.4% and uniparity was practically dominant one. A proportion of 26.9% of An. cruzii and 12.0% of An. bellator were found to be uniparous. Only three outdoor females of the former species (0.1% showed biparity. Parity of An. cruzii was higher in females caught outdoors than in those caught indoors. Nevertheless, 497 nulliparous females examined (417 cruzii and 80 bellator had ovaries that had advanced to Christophers and Mer stages III to V. These results imply that these females had already practised hematophagy. Relating these results to those from the parous females, a high statistical significance was found, leading to the conclusion that gonothophic discordance is a common pattern among these anophelines. Further, these results obtained with human bait catches strongly suggest that nearly 38.0% of these host-seeking females had already taken at least one previous blood-meal. So it is possible that enough time could thus be available for the plasmodian development in the vectors.Relata-se os resultados obtidos em coletas regulares de Anopheles cruzii e An. bellator, mediante o emprego de isca humana e por ocasião do crepúsculo vespertino. Objetivou-se, precipuamente, conhecer a paridade de populações dessas espécies, quando em plena tentativa hematófaga, tanto no ambiente intra como peridomiciliar. As coletas foram levadas a

  8. Hemocyte differentiation mediates innate immune memory in Anopheles gambiae mosquitoes.

    Science.gov (United States)

    Rodrigues, Janneth; Brayner, Fábio André; Alves, Luiz Carlos; Dixit, Rajnikant; Barillas-Mury, Carolina

    2010-09-10

    Mosquito midgut invasion by ookinetes of the malaria parasite Plasmodium disrupts the barriers that normally prevent the gut microbiota from coming in direct contact with epithelial cells. This triggers a long-lived response characterized by increased abundance of granulocytes, a subpopulation of hemocytes that circulates in the insect's hemocoel, and enhanced immunity to bacteria that indirectly reduces survival of Plasmodium parasites upon reinfection. In mosquitoes, differentiation of hemocytes was necessary and sufficient to confer innate immune memory. PMID:20829487

  9. Hemocyte Differentiation Mediates Innate Immune Memory in Anopheles gambiae Mosquitoes

    OpenAIRE

    Rodrigues, Janneth; Brayner, Fábio André; Alves, Luiz Carlos; Dixit, Rajnikant; Barillas-Mury, Carolina

    2010-01-01

    Mosquito midgut invasion by ookinetes of the malaria parasite Plasmodium disrupts the barriers that normally prevent the gut microbiota from coming in direct contact with epithelial cells. This triggers a long-lived response characterized by increased abundance of granulocytes, a subpopulation of hemocytes, circulating in the insect’s hemocoel, and enhanced immunity to bacteria that indirectly reduces survival of Plasmodium parasites upon reinfection. In mosquitoes, differentiation of hemocyt...

  10. Hemocyte Differentiation Mediates Innate Immune Memory in Anopheles gambiae Mosquitoes

    Science.gov (United States)

    Rodrigues, Janneth; Brayner, Fábio André; Alves, Luiz Carlos; Dixit, Rajnikant; Barillas-Mury, Carolina

    2012-01-01

    Mosquito midgut invasion by ookinetes of the malaria parasite Plasmodium disrupts the barriers that normally prevent the gut microbiota from coming in direct contact with epithelial cells. This triggers a long-lived response characterized by increased abundance of granulocytes, a subpopulation of hemocytes, circulating in the insect’s hemocoel, and enhanced immunity to bacteria that indirectly reduces survival of Plasmodium parasites upon reinfection. In mosquitoes, differentiation of hemocytes was necessary and sufficient to confer innate immune memory. PMID:20829487

  11. Larvicidal efficacy of monoterpenes against the larvae of Anopheles gambiae

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    Eliningaya J. Kweka

    2016-04-01

    Conclusions: The data obtained in this study suggest that all evaluated monoterpenes, especially (−-perillyl alcohol, have remarkable larvicidal effects and may be considered as potential sources for the development of suitable natural larvicides for mosquito management programs. Further small-scale field trials should be conducted.

  12. The role of cow urine in the oviposition site preference of culicine and Anopheles mosquitoes

    OpenAIRE

    Kweka Eliningaya J; Owino Eunice A; Mwang'onde Beda J; Mahande Aneth M; Nyindo Mramba; Mosha Franklin

    2011-01-01

    Abstract Background Chemical and behavioural ecology of mosquitoes plays an important role in the development of chemical cue based vector control. To date, studies available have focused on evaluating mosquito attractants and repellents of synthetic and human origins. This study, however, was aimed at seasonal evaluation of the efficiency of cow urine in producing oviposition cues to Anopheles gambiae s.l. and Culex quinquefasciatus in both laboratory and field conditions. Methods Ovipositio...

  13. [The Anopheles fauna and the transmission of human malaria in Kinshasa (Zaire)].

    Science.gov (United States)

    Karch, S; Asidi, N; Manzambi, Z M; Salaun, J J

    1992-01-01

    A longitudinal epidemiological study of malaria and its vectors was conducted in Kinshasa. 264 night-bite collections on human bait (1,056 man nights) and 384 collections of the house-resting fauna were carried out from April 1989 to October 1990. The anophelian fauna was identified and inventoried, 7 Anopheles species were found: Anopheles gambiae, An. funestus, An. paludis, An. hancocki, An. counstani, An. brunnipes, and An. nili. A single species, An. gambiae s. l. is responsible for the transmission of malaria, it represents 93.27% of the anopheline fauna. The average number of anophele bites man day was 16.28 bites/man/night, it varied between 1 b/m/n in urban area to 26.05 b/m/n in semi-rural area. The average of the sporozoite index for An. gambiae was 3.3%, but it varied from 0% in the urban area to 6.52% in the semi-rural area. The entomological inoculation rate (h) was 197 infective bites per year. This rate fluctuated from 1 infective bite each 128 nights in urban area to 1.7 infective night-bite in semi-rural area. Other epidemiological index were also determined: the level of daily survival rate (p = 8.75 days), the vectorial capacity of 17.97 and the Macdonald's stability 3.5 bites on man taken by a vector during its entire lifetime.

  14. Mermithid nematodes found in adult Anopheles from southeastern Senegal

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    Kobylinski Kevin C

    2012-06-01

    Full Text Available Abstract Background Over two dozen mermithid nematodes have been described parasitizing mosquitoes worldwide, however, only two species were found in Africa. Mermithid nematodes kill their mosquito host upon emergence, which suggests that they could be developed as biological control agents of mosquitoes. Both Romanomermis culicivorax and Romanomermis iyengari have been reared for mass release to control numerous Anopheles species vector populations, and in one instance this may have led to reduced malaria prevalence in a human population. Methods Anopheles mosquitoes were collected during a malaria study in southeastern Senegal. Two different adult blood fed mosquitoes had a single mermithid nematode emerge from their anus while they were being held post-capture. Primers from the 18 S rDNA were developed to sequence nematode DNA and screen mosquitoes for mermithid DNA. 18 S rDNA from the Senegalese mermithid and other mermithid entries in GenBank were used to create a Maximum Parsimony tree of the Mermithidae family. Results The mermithid was present in 1.8% (10/551 of the sampled adult Anopheles species in our study area. The mermithid was found in An. gambiae s.s., An. funestus, and An. rufipes from the villages of Ndebou, Boundoucondi, and Damboucoye. Maximum parsimony analysis confirmed that the nematode parasites found in Anopheles were indeed mermithid parasites, and of the mermithid sequences available in GenBank, they are most closely related to Strelkovimermis spiculatus. Conclusions To our knowledge, this is the first report of mermithids from adult Anopheles mosquitoes in Senegal. The mermithid appears to infect Anopheles mosquitoes that develop in diverse larval habitats. Although maximum parsimony analysis determined the mermithid was closely related to Strelkovimermis spiculatus, several characteristics of the mermithid were more similar to the Empidomermis genus. Future mermithid isolations will hopefully allow: formal

  15. Resistance Status of the Malaria Vector Mosquitoes, Anopheles stephensi and Anopheles subpictus Towards Adulticides and Larvicides in Arid and Semi-Arid Areas of India

    OpenAIRE

    Tikar, S. N.; M J Mendki; Sharma, A K; D. Sukumaran; Veer, Vijay; Prakash, Shri; Parashar, B. D.

    2011-01-01

    Susceptibility studies of malaria vectors Anopheles stephensi Liston (Diptera: Culicidae) and An. subpictus Grassi collected during 2004–2007 from various locations of Arid and Semi-Arid Zone of India were conducted by adulticide bioassay of DDT, malathion, deltamethrin and larvicide bioassay of fenthion, temephos, chlorpyriphos and malathion using diagnostic doses. Both species from all locations exhibited variable resistance to DDT and malathion from majority of location. Adults of both the...

  16. Sibling species of the Anopheles funestus group, and their infection with malaria and lymphatic filarial parasites, in archived and newly collected specimens from northeastern Tanzania

    DEFF Research Database (Denmark)

    Derua, Yahya A; Alifrangis, Michael; Magesa, Stephen M;

    2015-01-01

    , and Anopheles leesoni, with the first being by far the most common (overall 94.4%). When comparing archived specimens from 2005-2007 to those from 2008-2012, a small but statistically significant decrease in proportion of An. funestus s.s. was noted, but otherwise observed temporal changes in sibling species......BACKGROUND: Studies on the East African coast have shown a recent dramatic decline in malaria vector density and change in composition of sibling species of the Anopheles gambiae complex, paralleled by a major decline in malaria incidence. In order to better understand the ongoing changes in vector......-parasite dynamics in the area, and to allow for appropriate adjustment of control activities, the present study examined the composition, and malaria and lymphatic filarial infection, of sibling species of the Anopheles funestus group. Similar to the An. gambiae complex, the An. funestus group contains important...

  17. Biología de Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae en la Costa Pacifica de Colombia: IV - Estructura etárea y transmisión de malaria Biologia do Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae na Costa do Pacífico Colombiano: IV - Estrutura etária e transmissão da malária Biology of Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae on the Pacific Coast of Colombia: IV - Age structure and malaria transmission

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    César Murillo B.

    1989-10-01

    Full Text Available Con el fin de determinar la dinamica de transmisión de malaria en el poblado de Charambirá (Chocó, Colombia, se estudió la estructura etárea de Anopheles neivai (reconocido vector en la Costa Pacífica con base en su estado gonadotrófico. Se recolectaron mosquitos intradomiciliarmente durante el crepúsculo vespertino con cebos humanos y aspiradores bucales. Los mosquitos recolectados se mantuvieron en cajas cilíndricas de cartón con papel humedecido y dispensadores alimenticios hasta su disección al día siguiente. De los 200 mosquitos disecados entre septiembre y octubre de 1986,68 (34% presentaban huellas de menos de tres oviposturas y los restantes (66% habían efectuado al menos tres oviposturas. La diferencia entre el primer grupo considerado como mosquitos "no infectivos" y el segundo considerado como los "potencialmente infectivos" fue altamente significativa (X² = 10,68; P = 0,001. El 1,5% de A. neivai estudiados presentaban huellas correspondientes a 10 oviposturas mostrando una marcada longevidad y múltiples alimentaciones sanguíneas. Los resultados sugieren que existe un alto riesgo de contraer malaria en Charambirá durante el crepúsculo vespertino.Com o intuito de determinar a dinâmica de transmissão da malária no povoado de Charambirá (Chocó, Colômbia, foi estudada a estrutura etária do mosquito Anopheles neivai (reconhecido vetor da Costa Pacífica com base em seu estado gonadotrófico. As coletas foram realizadas intradomiciliarmente no crepúsculo vespertino com isca humana e auxílio de tubos de sucção manual. Os mosquitos coletados foram mantidos em caixas cilíndricas de papelão, contendo papel umidecido e alimento. Dos 200 mosquitos dissecados entre setembro e outubro de 1986, 68 (34% apresentaram vestígios de menos de três oviposturas (oviposições e os demais (66% apresentavam, pelo menos, três ovipoturas. A diferença entre o primeiro grupo considerado como mosquitos "não infectantes" e o

  18. Taxonomy Icon Data: Anopheles stephensi [Taxonomy Icon

    Lifescience Database Archive (English)

    Full Text Available Anopheles stephensi Anopheles stephensi Arthropoda Anopheles_stephensi_L.png Anopheles_stephen...si_NL.png Anopheles_stephensi_S.png Anopheles_stephensi_NS.png http://biosciencedbc.jp/taxonomy_i...con/icon.cgi?i=Anopheles+stephensi&t=L http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Anopheles+stephensi&...t=NL http://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Anopheles+stephensi&t=S htt...p://biosciencedbc.jp/taxonomy_icon/icon.cgi?i=Anopheles+stephensi&t=NS http://togodb.biosciencedbc.jp/togodb/view/taxonomy_icon_comment_en?species_id=149 ...

  19. Observations on the swarming and mating behaviour of Anopheles funestus from southern Mozambique

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    Thompson R

    2003-02-01

    Full Text Available Abstract Background Control of malaria by the release of genetically modified mosquitoes refractory to transmission is now becoming a possibility. In many areas of Africa, Anopheles gambiae is found together with an equally important vector, An. funestus. Given their sympatry and the likelihood of a similar mating period some aspects of the mating behaviour of An. gambiae s.l. and An. funestus are likely to differ. We therefore attempted to characterise the swarming behaviour of An. funestus and to determine if any aspects of the observed behaviour differed from that recorded for the M form of An. gambiae from São Tomé. Methods In March – May 2002 the swarming, mating, house exiting and resting behaviour of Anopheles funestus was studied by direct observation in Mozambique. Swarming males and insects in copula were collected by sweep net. Wing lengths of males collected resting, exiting houses, swarming and mating were measured and the wingbeat frequency distribution of individual insects, in free flight confined inside netting covered paper cups, was also determined. Results Mono-specific swarms occurred at sunset in relatively open areas close to houses used for resting. Mating pairs were seen 11 ± 3.7 min after the start of swarming. The number of total pairs observed being inversely proportional to the time difference between the start of swarming and the first pairing. The great majority of females mated before feeding. Male or female size did not appear to affect mating success or other behaviours. During the study, ambient temperatures decreased and female, but not male, wing size increased. At 516 Hz, the flight tone of female An. funestus was similar to the 497 Hz of the local An. gambiae. Males dispersed if light or dark artificial horizontal markers were placed underneath naturally occurring swarms. Conclusion Differential response to markers would be sufficient for swarming in An. funestus and An. gambiae s.l. to occur in

  20. Mosquito repellent action of Blumea lacera (Asteraceae) against Anopheles stephensi and Culex quinquefasciatus.

    OpenAIRE

    Singh, S.P.; MITTAL, P.K.

    2014-01-01

    Petroleum ether extract of Blumea lacera was screened under laboratory conditions for repellent activity against mosquito vector Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae). The repellent activity of Blumea lacera extract was tested against mosquitoes in comparison with the DEET, which was used as a positive control. Results obtained from the laboratory experiment showed that the extract was effective against mosquito vectors even at a low dose. A direct rel...

  1. Biologia de Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae en la costa Pacífica de Colombia: I. Fluctuación de la población larval y características de sus criaderos Biologia de Anopheles (Kerteszia neivai H., D. & K,, 1913 (Diptera: Culicidae na costa do Pacífico Colombiano: I. Flutuação da população larval e características dos seus berçários (criadouros Biology of Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae on the Pacific coast of Colombia: I. Fluctuation of larval population and characteristics of the breeding places

    Directory of Open Access Journals (Sweden)

    César Murillo B.

    1988-04-01

    Full Text Available En Charambirá, localidad del município de Istmina (Chocó en la costa Pacífica de Colombia, donde existe un problema de malaria endémica, se estudió la fluctuación de la población larval y las características de los criaderos de Anopheles (Kerteszia neivai, especie considerada como posible vectora de malaria en esa región del país. La vegetación circundante fue dividida en tres estratos de acuerdo al grado de cobertura foliar. Se demarcaron cuadrantes de 100 m² en cada estrato para el muestreo de plantas epífitas de la família Bromeliaceae, en las cuales se acumula agua que sirve como criadero para esta especie de anofelino. Se tomaron datos de temperatura, pH y volumen del agua contenida en cada bromelia. El mayor número de larvas se detectó en el estrato 1 (manglar a una altitud inferior a 4 m, pero no se encontró evidencia significativa de estratificación vertical de la población larval de A. neivai hasta los 8 m. Se evidenció una correlación lineal positiva entre el número de larvas y el volumen de agua contenida en cada bromelia; por otra parte se observó también una correlación directa entre la precipitación mensual acumulada y la fluctuación poblacional de esta especie. Los índices larvales mas altos se observaron entre los meses de marzo a abril y de julio a agosto. La mortalidad larval fue alta en el primer estadío (43,5% y solo un 23,7% sobrevivió hasta el cuarto.Em Charambirá, localidade do Município de Istmina (Chocó, na costa do Pacífico Colombiano, localidade endêmica de malária, estudou-se a flutuação da população larval e as características dos locais de criação do Anopheles (Kerteszia neivai, espécie considerada como possível vector da malária naquela região do país. A vegetação circundante foi dividida em três estratos segundo seu grau de cobertura foliar. Demarcaram-se quadrantes de 100 m² para a amostragem de plantas epífitas da família Bromeliaceae, nas quais se

  2. Biologia de Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae en la costa Pacífica de Colombia: II. Fluctuación de la población adulta Biologia do Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae na costa do Pacífico Colombiano: II. Flutuação da população adulta Biology of Anopheles (Kerteszia neivai H., D. & K., 1913 (Diptera: Culicidae on the Pacific Coast of Colombia: II. Adult population fluctuation

    Directory of Open Access Journals (Sweden)

    Rodrigo Astaiza V.

    1988-04-01

    Full Text Available En la localidad de Charambirá, situada en el município de Istmina (Chocó en la costa Pacífica de Colombia, se presenta un problema de malaria endémica asociada con grandes poblaciones de mosquitos Anopheles del subgénero Kerteszia (A. neivai. Estos se crían en las colecciones de agua formadas por plantas epífitas de la família Bromeliaceae, que son muy abundantes en los árboles de mangle. En esa localidad se estudiaron: fluctuación poblacional y algunos aspectos ecológicos y etológicos de los adultos de esa especie de mosquito, con el objetivo de determinar su papel en la transmisión de malaria. Todos los mosquitos fueron colectados por cebos humanos. Los picos máximos de actividad ocurrieron en las horas crepusculares de la mañana y de la tarde (5:30 a 6:30 y 18:00 a 19:00 horas. En el período nocturno la actividad intradomiciliar fue baja y no hubo ninguna durante el día. El estado gonadotrófico de las hembras colectadas durante nos picos de actividad, mostró la existencia de dos poblaciones de mosquitos: una, en su mayoría individuos jóvenes que buscan su alimento en las horas de la tarde y la otra compuesta por individuos mas longevos, que buscan su alimento en la mañana. El estudio de la variación estacional mostró que las poblaciones son bajas en los meses de poca precipitación, pero a medida que aumenta el índice pluviométrico, aumenta el número de mosquitos. Se discute la relación que existe entre la presencia de los mosquitos y la prevalencia de malaria humana; se sugiere que A. neivai puede ser el responsable de la transmisión de malaria en la zona estudiada.Na localidade de Charambirá, situada no Municipio de Istmina (Chocó, na costa do Pacífico Colombiano, ocorre um tipo de malária endêmica associada com grandes populações de Anopheles do subgênero Kerteszia (A. neivai. Estes criam-se em coleções de água formadas por plantas epífitas pertencentes à família Bromeliaceae, as quais são muito

  3. Pteridine fluorescence for age determination of Anopheles mosquitoes.

    Science.gov (United States)

    Wu, D; Lehane, M J

    1999-02-01

    The age structure of mosquito populations is of great relevance to understanding the dynamics of disease transmission and in monitoring the success of control operations. Unfortunately, the ovarian dissection methods currently available for determining the age of adult mosquitoes are technically difficult, slow and may be of limited value, because the proportion of diagnostic ovarioles in the ovary declines with age. By means of reversed-phase HPLC this study investigated the malaria vectors Anopheles gambiae and An. stephensi to see if changes in fluorescent pteridine pigments, which have been used in other insects to determine the age of field-caught individuals, may be useful for age determination in mosquitoes. Whole body fluorescence was inversely proportional to age (P 91%) up to 30 days postemergence, with the regression values: y = 40580-706x for An. gambiae, and y = 52896-681x for An. stephensi. In both species the main pteridines were 6-biopterin, pterin-6-carboxylic acid and an unidentified fluorescent compound. An. gambiae had only 50-70% as much fluorescence as An. stephensi, and fluorescent compounds were relatively more concentrated in the head than in the thorax (ratios 1:0.8 An. gambiae; 1:0.5 An. stephensi). The results of this laboratory study are encouraging. It seems feasible that this simpler and faster technique of fluorescence quantification could yield results of equivalent accuracy to the interpretation of ovarian dissection. A double-blind field trial comparing the accuracy of this technique to marked, released and recaptured mosquitoes is required to test the usefulness of the pteridine method in the field. PMID:10194749

  4. Prevalence of anopheles (Diptera: Culicidae) during sunset in areas of the Itaipu Hydroelectric Power Plant in Guaira County, State of Parana, Brazil; Prevalencia de anofelinos (Diptera: Culicidae) no crepusculo vespertino em areas da Usina Hidreletrica de Itaipu, no Municipio de Guaira, Estado do Parana, Brasil

    Energy Technology Data Exchange (ETDEWEB)

    Guimaraes, Anthony Erico; Mello, Rubens Pinto de; Lopes, Catarina Macedo; Alencar, Jeronimo; Gentile, Carla [Fundacao Inst. Oswaldo Cruz (FIOCRUZ), Rio de Janeiro, RJ (Brazil). Dept. de Entomologia. Lab. de Diptera

    1997-11-01

    Systematic collections of anophelines were conducted from November 1995 to August 1995 from 18:00 to 20:00 hr using Shannon traps and human-bait along the lake margin which forms the Itaipu Hydroelectric Reservoir, State of Parana, Brazil. Species prevalence was studied at 15 min intervals. Anopheles albitarsis sensu latu and An. galvaoi, were the most frequently collected mosquitoes. All Anopheles species populations peaked between 18:45 and 19:30 hr. The observations illustrate the existence of haematophagic activity cycle during the early evening hours: exogenous stimulus (the beginning of sunset) Shannon trap (light attraction) human bait (haematophagy) rest and digestion exogenous stimulus Shannon trap or surrounding vegetation. The greater abundance of An. albitarsis collected in human-bait and Shannon trap suggests it may be a potential malaria vector in the region. (author) 20 refs.

  5. "Use of Random Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR and ITS2 PCR assays for differentiation of populations and putative sibling species of Anopheles fluviatilis (Diptera: Culicidae in Iran"

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    SR Naddaf Dezfouli

    2002-09-01

    Full Text Available Anopheles fluviatilis complex is known to be a vector of malaria in Iran. Since mosquitoes of this species cover a wide geographical range in Iran, they might have evolved into different separated populations. Random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR assay was used to differentiate geographic populations of this species. DNA was extracted from individual mosquitoes from 8 localities in 4 south and southeast provinces and amplified in PCR reactions using 18 single primers of arbitrary nucleotide sequence. Results of RAPD-PCR showed that Kazeroun populations could simply be differentiated from other populations using a diagnostic fragment amplified with primer UBC-306. But other populations could not be differentiated either visually or by means of statistical analysis. Moreover ITS2 fragments of some selected specimens were amplified using a pair of universal primer and sequenced as a key standard for detection of putative sibling species. Sequence analysis of the ITS2 fragments revealed a very high (100% homology among the populations. These findings are crucial in epidemiological studies concerning relatedness of geographic populations and vector movement in the region. Results of RAPD-PCR and ITS2 analysis suggest that this taxon in Iran comprises of only one species with a low genetic variation among geographic populations.

  6. Additional selection for insecticide resistance in urban malaria vectors: DDT resistance in Anopheles arabiensis from Bobo-Dioulasso, Burkina Faso.

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    Christopher M Jones

    Full Text Available In the city of Bobo-Dioulasso in Burkina Faso, Anopheles arabiensis has superseded Anopheles gambiae s.s. as the major malaria vector and the larvae are found in highly polluted habitats normally considered unsuitable for Anopheles mosquitoes. Here we show that An. gambiae s.l. adults emerging from a highly polluted site in the city centre (Dioulassoba have a high prevalence of DDT resistance (percentage mortality after exposure to diagnostic dose=65.8% in the dry season and 70.4% in the rainy season, respectively. An investigation into the mechanisms responsible found an unexpectedly high frequency of the 1014S kdr mutation (allele frequency=0.4, which is found at very low frequencies in An. arabiensis in the surrounding rural areas, and an increase in transcript levels of several detoxification genes, notably from the glutathione transferase and cytochrome P450 gene families. A number of ABC transporter genes were also expressed at elevated levels in the DDT resistant An. arabiensis. Unplanned urbanisation provides numerous breeding grounds for mosquitoes. The finding that Anopheles mosquitoes adapted to these urban breeding sites have a high prevalence of insecticide resistance has important implications for our understanding of the selective forces responsible for the rapid spread of insecticide resistant populations of malaria vectors in Africa.

  7. Nota sobre culicídeos (Diptera: Culicidae da bacia do rio Purus, Acre, Amazônia (Brasil

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    Delsio Natal

    1992-04-01

    Full Text Available Foram feitas coletas de mosquito (Diptera: Culicidae na área do projeto de Colonização Pedro Peixoto, no Estado do Acre, Brasil. Obteve-se um total de 4.588 exemplares pertencentes a 53 espécies ou grupos. Salienta-se a ocorrência de Anopheles (Nyssorhynchus oswaldoi.

  8. Evaluation of a eucalyptus-based repellent against Anopheles spp. in Tanzania.

    Science.gov (United States)

    Trigg, J K

    1996-06-01

    A eucalyptus-based insect repellent (PMD) with the principal active ingredient p-menthane-3,8-diol was evaluated in the field in comparison with deet. In human landing catches in Tanzania, 3 formulations of PMD were tested against Anopheles gambiae and An. funestus. Repellents, applied to the legs and feet at doses chosen as used in practice, gave complete protection from biting for between 6 and 7.75 h, depending upon the formulation type, with no significant difference between PMD and deet in terms of efficacy and duration of protection. PMID:8827599

  9. α-Humulene and β-elemene from Syzygium zeylanicum (Myrtaceae) essential oil: highly effective and eco-friendly larvicides against Anopheles subpictus, Aedes albopictus, and Culex tritaeniorhynchus (Diptera: Culicidae).

    Science.gov (United States)

    Govindarajan, Marimuthu; Benelli, Giovanni

    2016-07-01

    Mosquitoes transmit serious pathogens and parasites to humans and animals, including malaria, dengue, Japanese encephalitis and filariasis. The extensive use of chemical pesticides leads to the development of resistance in mosquito vector populations and serious non-target effects on human health and the environment. Myrtaceae plants can be a useful reservoir of natural products effective against Culicidae young instars. In this research, we evaluated the mosquitocidal potential of the essential oil (EO) from Syzygium zeylanicum leaves against larvae of three mosquitoes of medical and veterinary importance, the malaria vector Anopheles subpictus, the dengue vector Aedes albopictus, and the Japanese encephalitis vector Culex tritaeniorhynchus. The chemical composition of the EO was analyzed by gas chromatography-mass spectroscopy. GC-MS revealed that the S. zeylanicum EO contained at least 18 compounds. The major chemical components were α-humulene (37.8.5 %) and β-elemene (10.7 %). The EO had a significant toxic effect against early third-stage larvae of An. subpictus, Ae. albopictus, and Cx. tritaeniorhynchus, with LC50 values of 83.11, 90.45, and 97.96 μg/ml, respectively. The two major constituents extracted from the S. zeylanicum EO were tested individually for acute toxicity against larvae of the three mosquito vectors. α-Humulene and β-elemene appeared highly effective against An. subpictus (LC50 = 6.19 and 10.26 μg/ml, respectively), followed by Ae. albopictus (LC50 = 6.86 and 11.15 μg/ml) and Cx. tritaeniorhynchus (LC50 = 7.39 and 12.05 μg/ml). Furthermore, the EO and its major components was safe towards the non-target fish Gambusia affinis; LC50 values were 20374.26, 1024.95, and 2073.18 μg/ml, respectively for EO, α-humulene and β-elemene. Overall, this study highlighted that the acute toxicity of S. zeylanicum EO towards mosquito larvae was mainly due to the presence of α-humulene and β-elemene. Furthermore, we pointed

  10. Sampling of An.gambiae s.s mosquitoes using Limburger cheese, heat and moisture as baits in a homemade trap

    Science.gov (United States)

    2011-01-01

    Background Ample evidence has shown that odour baited traps are likely to provide an objective monitoring tool for the host-seeking fraction of mosquito vectors of diseases like malaria and bancroftian filariasis. Such traps could eventually become part of primary healthcare systems used to study the vector biology and epidemiology of mosquito-borne diseases. I hereby, report a study that sampled Anopheles gambiae sensu stricto mosquitoes in a screen house using a homemade trap baited with a combination of Limburger cheese and moisture, Limburger cheese and heat, or Limburger cheese, moisture and heat. Findings Tests on the efficacy of the developed trap to sample An. gambiae s.s, mosquitoes using Limburger cheese, moisture and heat as baits were carried out in a screen house measuring 11.4 × 7.1 × 2.8 m. The studies were done in three phases. In the first phase the efficacy of the trap to sample An. gambiae s.s. using odour and moisture was tested. The second phase was to test the efficacy of the trap to sample An. gambiae s.s. using Limburger cheese and heat. In the third phase a combination of Limburger cheese, moisture and heat was tested. Tests were carried out for 27 consecutive nights. The designed trap collected a total of 59 An. gambiae s.s. in three trials. The trap baited with Limburger cheese and moisture collected 7 An. gambiae s.s in 7 days. The mean catch per day was 1. The trap baited with Limburger cheese and heat collected zero An. gambiae s.s in 11 days. The mean catch per day was therefore 0. The trap baited with Limburger cheese, moisture and heat collected 52 mosquitoes in 27 days and the mean catch was 1.93. Conclusions This study indicates that a non-electric fan driven trap baited with a combination of Limburger cheese, heat and moisture has a potential as an effective sampling tool for the malaria vector, Anopheles gambiae s.s. However, further optimization studies would be necessary. PMID:21835032

  11. Feeding and indoor resting behaviour of the mosquito Anopheles longipalpis in an area of hyperendemic malaria transmission in southern Zambia

    OpenAIRE

    Kent, R.J.; Coetzee, M.; Mharakurwa, S.; Norris, D. E.

    2006-01-01

    Anopheles longipalpis (Theobald) (Diptera: Culicidae) is a predominantly zoophilic mosquito that has not been implicated in malaria transmission. However, this species was collected indoors with An. funestus s.l. in southern Zambia, where transmission of Plasmodium falciparum is hyperendemic, and we initially misidentified it morphologically and molecularly as An. funestus s.l. The indoor resting density and blood-feeding behaviour of An. longipalpis were investigated during the 2004 – 05 and...

  12. [Mosquitoes (Diptera, Culicidae) of Smir marshes (northwest of Morocco): inventory and biotypology].

    Science.gov (United States)

    El Joubari, M; Louah, A; Himmi, O

    2014-02-01

    The Smir marshes are a favorable environment for the growth of many mosquitoes (Diptera, Culicidae). The inventory of Culicidae species reveals 14 species, is 33% of the species of Morocco, distributed in four genera: Culex, Culiseta, Ochlerotatus and Anopheles (with 5, 2, 5 and 2 species respectively) which Anopheles labranchiae, vector of the agent of the malaria in Morocco until 2004. In this study, we investigated the spatiotemporal mesological affinities and we tried to explain the biotypology of mosquito populations of the site. These analyzes revealed several groups of stations and species according to various parameters, especially salinity.

  13. Systematics of mosquito disease vectors (Diptera, Culicidae): impact of molecular biology and cladistic analysis.

    Science.gov (United States)

    Munstermann, L E; Conn, J E

    1997-01-01

    The field of medical entomology, by nature of its association with problems of human health, has been conservative in its application of molecular and computer technologies to systematic research. Recently, however, these methods have opened new interpretations for systematics of disease vectors. Medically important insects, particularly mosquitoes, are among those more thoroughly described by conventional taxonomy, and thereby provide a secure framework for testing congruencies with molecular data. In turn, molecular investigations have provided a stimulus to vector systematics in the discovery and delineation of cryptic species complexes, as well as providing new perspectives on relationships at higher taxonomic divisions. In this review, examples involving cladistic analysis, cytogenetics--in situ hybridization, isoenzymes, DNA sequencing, and restriction fragment polymorphism are drawn from the following taxa: Aedes communis; Aedes (Ochlerotatus) group G; Aedes (Stegomyia) species including A. aegypti, A. albopictus, and A. scutellaris group; Anopheles albitarsis, Anopheles dirus, Anopheles gambiae, Anopheles nuneztovari, Anopheles pseudopunctipennis, and Anopheles punctulatus groups; Culex pipiens and the Culex subgenus Melanoconion; and the tribe Sabethini.

  14. Genome-Wide Divergence in the West-African Malaria Vector Anopheles melas

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    Kevin C. Deitz

    2016-09-01

    Full Text Available Anopheles melas is a member of the recently diverged An. gambiae species complex, a model for speciation studies, and is a locally important malaria vector along the West-African coast where it breeds in brackish water. A recent population genetic study of An. melas revealed species-level genetic differentiation between three population clusters. An. melas West extends from The Gambia to the village of Tiko, Cameroon. The other mainland cluster, An. melas South, extends from the southern Cameroonian village of Ipono to Angola. Bioko Island, Equatorial Guinea An. melas populations are genetically isolated from mainland populations. To examine how genetic differentiation between these An. melas forms is distributed across their genomes, we conducted a genome-wide analysis of genetic differentiation and selection using whole genome sequencing data of pooled individuals (Pool-seq from a representative population of each cluster. The An. melas forms exhibit high levels of genetic differentiation throughout their genomes, including the presence of numerous fixed differences between clusters. Although the level of divergence between the clusters is on a par with that of other species within the An. gambiae complex, patterns of genome-wide divergence and diversity do not provide evidence for the presence of pre- and/or postmating isolating mechanisms in the form of speciation islands. These results are consistent with an allopatric divergence process with little or no introgression.

  15. Analysis of two novel midgut-specific promoters driving transgene expression in Anopheles stephensi mosquitoes.

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    Tony Nolan

    Full Text Available BACKGROUND: Tissue-specific promoters controlling the expression of transgenes in Anopheles mosquitoes represent a valuable tool both for studying the interaction between these malaria vectors and the Plasmodium parasites they transmit and for novel malaria control strategies based on developing Plasmodium-refractory mosquitoes by expressing anti-parasitic genes. With this aim we have studied the promoter regions of two genes from the most important malaria vector, Anopheles gambiae, whose expression is strongly induced upon blood feeding. RESULTS: We analysed the A. gambiae Antryp1 and G12 genes, which we have shown to be midgut-specific and maximally expressed at 24 hours post-bloodmeal (PBM. Antryp1, required for bloodmeal digestion, encodes one member of a family of 7 trypsin genes. The G12 gene, of unknown function, was previously identified in our laboratory in a screen for genes induced in response to a bloodmeal. We fused 1.1 kb of the upstream regions containing the putative promoter of these genes to reporter genes and transformed these into the Indian malaria vector A. stephensi to see if we could recapitulate the expression pattern of the endogenous genes. Both the Antryp1 and G12 upstream regions were able to drive female-predominant, midgut-specific expression in transgenic mosquitoes. Expression of the Antryp1-driven reporter in transgenic A. stephensi lines was low, undetectable by northern blot analysis, and failed to fully match the induction kinetics of the endogenous Antryp1 gene in A. gambiae. This incomplete conservation of expression suggests either subtle differences in the transcriptional machinery between A. stephensi and A. gambiae or that the upstream region chosen lacked all the control elements. In contrast, the G12 upstream region was able to faithfully reproduce the expression profile of the endogenous A. gambiae gene, showing female midgut specificity in the adult mosquito and massive induction PBM, peaking at 24

  16. The susceptibility of five African Anopheles species to Anabaena PCC 7120 expressing Bacillus thuringiensis subsp. israelensis mosquitocidal cry genes

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    Ketseoglou Irene

    2012-10-01

    Full Text Available Abstract Background Malaria, one of the leading causes of death in Africa, is transmitted by the bite of an infected female Anopheles mosquito. Problems associated with the development of resistance to chemical insecticides and concerns about the non-target effects and persistence of chemical insecticides have prompted the development of environmentally friendly mosquito control agents. The aim of this study was to evaluate the larvicidal activity of a genetically engineered cyanobacterium, Anabaena PCC 7120#11, against five African Anopheles species in laboratory bioassays. Findings There were significant differences in the susceptibility of the anopheline species to PCC 7120#11. The ranking of the larvicidal activity of PCC 7120#11 against species in the An. gambiae complex was: An. merus An. arabiensis An. gambiae An. quadriannulatus, where 50. The LC50 of PCC 7120#11 against the important malaria vectors An. gambiae and An. arabiensis was 12.3 × 105 cells/ml and 8.10 × 105 cells/ml, respectively. PCC 7120#11 was not effective against An. funestus, with less than 50% mortality obtained at concentrations as high as 3.20 × 107 cells/ml. Conclusions PCC 7120#11 exhibited good larvicidal activity against larvae of the An. gambiae complex, but relatively weak larvicidal activity against An. funestus. The study has highlighted the importance of evaluating a novel mosquitocidal agent against a range of malaria vectors so as to obtain a clear understanding of the agent’s spectrum of activity and potential as a vector control agent.

  17. Risk factors for house-entry by malaria vectors in a rural town and satellite villages in The Gambia

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    Jasseh Momadou

    2008-01-01

    Full Text Available Abstract Background In the pre-intervention year of a randomized controlled trial investigating the protective effects of house screening against malaria-transmitting vectors, a multi-factorial risk factor analysis study was used to identify factors that influence mosquito house entry. Methods Mosquitoes were sampled using CDC light traps in 976 houses, each on one night, in Farafenni town and surrounding villages during the malaria-transmission season in The Gambia. Catches from individual houses were both (a left unadjusted and (b adjusted relative to the number of mosquitoes caught in four sentinel houses that were operated nightly throughout the period, to allow for night-to-night variation. Houses were characterized by location, architecture, human occupancy and their mosquito control activities, and the number and type of domestic animals within the compound. Results 106,536 mosquitoes were caught, of which 55% were Anopheles gambiae sensu lato, the major malaria vectors in the region. There were seven fold higher numbers of An. gambiae s.l. in the villages (geometric mean per trap night = 43.7, 95% confidence intervals, CIs = 39.5–48.4 than in Farafenni town (6.3, 5.7–7.2 and significant variation between residential blocks (p Conclusion This study demonstrates that the risk of malaria transmission is greatest in rural areas, where large numbers of people sleep in houses made of mud blocks, where the eaves are open, horses are not tethered nearby and where churai is not burnt at night. These factors need to be considered in the design and analysis of intervention studies designed to reduce malaria transmission in The Gambia and other parts of sub-Saharan Africa.

  18. Molasses as a source of carbon dioxide for attracting the malaria mosquitoes Anopheles gambiae and Anopheles funestus

    OpenAIRE

    Mweresa, C. K.; Omusula, P.; Otieno, B.; Loon, van, R.R.; Takken, W.; Mukabana, W.R.

    2014-01-01

    Background. Most odour baits for haematophagous arthropods contain carbon dioxide (CO2). The CO2 is sourced artificially from the fermentation of refined sugar (sucrose), dry ice, pressurized gas cylinders or propane. These sources of CO2 are neither cost-effective nor sustainable for use in remote areas of sub-Saharan Africa. In this study, molasses was evaluated as a potential substrate for producing CO2 used as bait for malaria mosquitoes. Methods. The attraction of laboratory-reared and w...

  19. Pyrethroid resistance in an Anopheles funestus population from Uganda.

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    John C Morgan

    Full Text Available BACKGROUND: The susceptibility status of Anopheles funestus to insecticides remains largely unknown in most parts of Africa because of the difficulty in rearing field-caught mosquitoes of this malaria vector. Here we report the susceptibility status of the An. funestus population from Tororo district in Uganda and a preliminary characterisation of the putative resistance mechanisms involved. METHODOLOGY/PRINCIPAL FINDINGS: A new forced egg laying technique used in this study significantly increased the numbers of field-caught females laying eggs and generated more than 4000 F1 adults. WHO bioassays indicated that An. funestus in Tororo is resistant to pyrethroids (62% mortality after 1 h exposure to 0.75% permethrin and 28% mortality to 0.05% deltamethrin. Suspected DDT resistance was also observed with 82% mortality. However this population is fully susceptible to bendiocarb (carbamate, malathion (organophosphate and dieldrin with 100% mortality observed after exposure to each of these insecticides. Sequencing of a fragment of the sodium channel gene containing the 1014 codon conferring pyrethroid/DDT resistance in An. gambiae did not detect the L1014F kdr mutation but a correlation between haplotypes and resistance phenotype was observed indicating that mutations in other exons may be conferring the knockdown resistance in this species. Biochemical assays suggest that resistance in this population is mediated by metabolic resistance with elevated level of GSTs, P450s and pNPA compared to a susceptible strain of Anopheles gambiae. RT-PCR further confirmed the involvement of P450s with a 12-fold over-expression of CYP6P9b in the Tororo population compared to the fully susceptible laboratory colony FANG. CONCLUSION: This study represents the first report of pyrethroid/DDT resistance in An. funestus from East Africa. With resistance already reported in southern and West Africa, this indicates that resistance in An. funestus may be more widespread

  20. Application of a qPCR assay in the investigation of susceptibility to malaria infection of the M and S molecular forms of An. gambiae s.s. in Cameroon.

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    Anne Boissière

    Full Text Available Plasmodium falciparum is the causative agent of malaria, a disease that kills almost one million persons each year, mainly in sub-Saharan Africa. P. falciparum is transmitted to the human host by the bite of an Anopheles female mosquito, and Anopheles gambiae sensus stricto is the most tremendous malaria vector in Africa, widespread throughout the afro-tropical belt. An. gambiae s.s. is subdivided into two distinct molecular forms, namely M and S forms. The two molecular forms are morphologically identical but they are distinct genetically, and differ by their distribution and their ecological preferences. The epidemiological importance of the two molecular forms in malaria transmission has been poorly investigated so far and gave distinct results in different areas. We have developed a real-time quantitative PCR (qPCR assay, and used it to detect P. falciparum at the oocyst stage in wild An. gambiae s.s. mosquitoes experimentally infected with natural isolates of parasites. Mosquitoes were collected at immature stages in sympatric and allopatric breeding sites and further infected at the adult stage. We next measured the infection prevalence and intensity in female mosquitoes using the qPCR assay and correlated the infection success with the mosquito molecular forms. Our results revealed different prevalence of infection between the M and S molecular forms of An. gambiae s.s. in Cameroon, for both sympatric and allopatric populations of mosquitoes. However, no difference in the infection intensity was observed. Thus, the distribution of the molecular forms of An. gambiae s.s. may impact on the malaria epidemiology, and it will be important to monitor the efficiency of malaria control interventions on the two M and S forms.

  1. Larvicidal activity of the leaf extracts of Spondias mombin Linn. (Anacardiaceae) from various solvents against malarial, dengue and filarial vector mosquitoes (Diptera: Culicidae)

    OpenAIRE

    Elijah Ajaegbu Eze; Simon Pierre Yinyang Danga; Festus Basden Chiedu Okoye

    2015-01-01

    Background & objectives: Aedes aegypti, Anopheles gambiae and Culex quinquefasciatus are vector mosquitoes of dengue, malaria, and filariasis, respectively. Since no vaccine is available to treat these diseases, the control of the main mosquito vectors is essential. As conventional insecticides have limited success, plants may be alternative larvicidal agents, since they contain a rich source of bioactive chemicals. The aim of this study was to evaluate the larvicidal activity of methanol cru...

  2. Robust and regulatory expression of defensin A gene driven by vitellogenin promoter in transgenic Anopheles stephensi

    Institute of Scientific and Technical Information of China (English)

    CHEN XiaoGuang; ZHANG YaJing; ZHENG XueLi; WANG ChunMei

    2007-01-01

    The use of genetically modified mosquitoes to reduce or replace field populations is a new strategy to control mosquito-borne diseases. The precondition of the implementation of this strategy is the ability to manipulate the genome of mosquitoes and to induce specific expression of the effector molecules driven by a suitable promoter. The objective of this study is to evaluate the expression of defensin A gene of Anopheles sinensis under the control of a vitellogenin promoter in transgenic Anopheles stephensi. The regulatory region of Anopheles gambiae vitellogenin was cloned and subcloned into transfer vector pSLFa consisting of an expression cassette with defensin A coding sequence. Then, the expression cassette was transferred into transformation vector pBac[3xP3-DsRedafm] using Asc I digestion. The recombinant plasmid DNA of pBac[3xP3DsRed-AgVgT2-DefA] and helper plasmid DNA of phsp-pBac were micro-injected into embryos of An. stephensi. The positive transgenic mosquitoes were screened by observing specific red fluorescence in the eyes of G1 larvae. Southern blot analysis showed that a single-copy transgene integrated into the genome of An. stephensi. RT-PCR analysis showed that the defensin A gene expressed specifically in fat bodies of female mosquitoes after a blood meal. Interestingly, the mRNA of defensin A is more stable compared with that of the endogenous vitellogenin gene. After multiple blood meals, the expression of defensin A appeared as a reducible and non-cycling type, a crucial feature for its anti-pathogen effect. From data above, we concluded that the regulatory function of the Vg promoter and the expression of defensin A gene were relatively conserved in different species of anopheles mosquitoes. These molecules could be used as candidates in the development of genetically modified mosquitoes.

  3. Elimination of lymphatic filariasis in the Gambia.

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    Maria P Rebollo

    2015-03-01

    Full Text Available The prevalence of Wuchereria bancrofti, which causes lymphatic filariasis (LF in The Gambia was among the highest in Africa in the 1950s. However, surveys conducted in 1975 and 1976 revealed a dramatic decline in LF endemicity in the absence of mass drug administration (MDA. The decline in prevalence was partly attributed to a significant reduction in mosquito density through the widespread use of insecticidal nets. Based on findings elsewhere that vector control alone can interrupt LF, we asked the question in 2013 whether the rapid scale up in the use of insecticidal nets in The Gambia had interrupted LF transmission.We present here the results of three independently designed filariasis surveys conducted over a period of 17 years (1997-2013, and involving over 6000 subjects in 21 districts across all administrative divisions in The Gambia. An immunochromatographic (ICT test was used to detect W. bancrofti antigen during all three surveys. In 2001, tests performed on stored samples collected between 1997 and 2000, in three divisions, failed to show positive individuals from two divisions that were previously highly endemic for LF, suggesting a decline towards extinction in some areas. Results of the second survey conducted in 2003 showed that LF was no longer endemic in 16 of 21 districts surveyed. The 2013 survey used a WHO recommended LF transmission verification tool involving 3180 6-7 year-olds attending 60 schools across the country. We demonstrated that transmission of W. bancrofti has been interrupted in all 21 districts.We conclude that LF transmission may have been interrupted in The Gambia through the extensive use of insecticidal nets for malaria control for decades. The growing evidence for the impact of malaria vector control activities on parasite transmission has been endorsed by WHO through a position statement in 2011 on integrated vector management to control malaria and LF.

  4. Elimination of lymphatic filariasis in the Gambia.

    OpenAIRE

    Rebollo, Maria P.; Sana Malang Sambou; Brent Thomas; Nana-Kwadwo Biritwum; Momodou C Jaye; Louise Kelly-Hope; Alba Gonzalez Escalada; Molyneux, David H; Bockarie, Moses J.

    2015-01-01

    Background The prevalence of Wuchereria bancrofti, which causes lymphatic filariasis (LF) in The Gambia was among the highest in Africa in the 1950s. However, surveys conducted in 1975 and 1976 revealed a dramatic decline in LF endemicity in the absence of mass drug administration (MDA). The decline in prevalence was partly attributed to a significant reduction in mosquito density through the widespread use of insecticidal nets. Based on findings elsewhere that vector control alone can interr...

  5. The Gambia : Country Procurement Issues Paper

    OpenAIRE

    World Bank

    2005-01-01

    This Country Procurement Issues Paper (CPIP), is an interim assessment of the public procurement system of The Gambia based on the self-evaluation conducted by the Government in early 2005. The CPIP follows-up the Country Procurement Assessment Review (CPAR) undertaken in 1998, which led to the preparation and implementation of the public procurement reform program. Since the 1998's review, progress has been made especially in the legal, regulatory, and institutional framework. In 2001, Parli...

  6. Agriculture and growth nexus in Gambia

    OpenAIRE

    Bukhari Sillah

    2013-01-01

    Agriculture is a major engine for the economic well-being of the Gambia. The successive governments from the colonial periods to the present have all recognized this importance of the agricultural sector but failed to do something that bear any fruit since government encouraged slash and burn technology and putting increasingly more people onto the land instead of increasing the yields per acre. More people than before do remain still on the land, but they are now poorer than their forefather...

  7. Agriculture and growth nexus in Gambia

    OpenAIRE

    Bukhari Sillah; Odior Simeon Ernest

    2014-01-01

    Agriculture is a major engine for the economic well-being of the Gambia. The successive governments from the colonial periods to the present have all recognized this importance of the agricultural sector but failed to do something that bear any fruit since government encouraged slash and burn technology and putting increasingly more people onto the land instead of increasing the yields per acre. More people than before do remain still on the land, but they are now poorer than their forefather...

  8. Agriculture and growth nexus in Gambia

    Directory of Open Access Journals (Sweden)

    Bukhari Sillah

    2013-12-01

    Full Text Available Agriculture is a major engine for the economic well-being of the Gambia. The successive governments from the colonial periods to the present have all recognized this importance of the agricultural sector but failed to do something that bear any fruit since government encouraged slash and burn technology and putting increasingly more people onto the land instead of increasing the yields per acre. More people than before do remain still on the land, but they are now poorer than their forefathers. Using autoregressive and vector error correction techniques to examine the growth-agriculture relation in the Gambia for the period from 1966 to 2009, it is found that the capital per worker is a significant and relevant factor input for the economic growth. The agricultural labor per acre is found to be irrelevant in both the short run and the long run analyses. The agricultural productivity measured as crop yields per acre is the most important variable for the economic growth in the Gambia. It boosts both the economic growth and the capital formation in the country. The agricultural policies should be focused on increasing the crop yields per acre not having more people back to the land. The processing, services and small manufacturing sectors should be developed and built into the agricultural policies in order to create redeployments for the agricultural labor surpluses.

  9. Modelling sterile insect technique to control the population of Anopheles gambiae

    OpenAIRE

    James E. Gentile; Rund, Samuel SC; Gregory R. Madey

    2015-01-01

    Background There is a renewed effort to develop novel malaria control strategies as even well-implemented existing malaria control tools may fail to block transmission in some regions. Currently, transgenic implementations of the sterile insect technique (SIT) such as the release of insects with a dominant lethal, homing endonuclease genes, or flightless mosquitoes are in development. These implementations involve the release of transgenic male mosquitoes whose matings with wild females produ...

  10. The accumulation of specific mRNAs following multiple blood meals in Anopheles gambiae

    OpenAIRE

    Xavier, Nirmala; Marinotti, Osvaldo; James, Anthony A

    2005-01-01

    One approach to genetic control of transmission of the parasites that cause human malaria is based on expressing effector genes in mosquitoes that disable the pathogens. Endogenous mosquito promoter and other cis-acting DNA sequences are needed to direct the optimal tissue-, stage- and sex-specific expression of the effector molecules. The mRNA accumulation profiles of eight different genes expressed specifically in the midgut, salivary glands or fat body tissues of the malaria vector, Anophe...

  11. Plasmodium falciparum infection increases Anopheles gambiae attraction to nectar sources and sugar uptake

    Science.gov (United States)

    Plasmodium parasites are known to manipulate the behaviour of their vectors so as to enhance their transmission. However, it is unknown if this vector manipulation also affects mosquito-plant interaction and sugar uptake. Dual-choice olfactometer and probing assays were used to study plant seeking b...

  12. Plasmodium sexual development and the role of Plasmepsin X in Plasmodium falciparum transmission to anopheles gambiae

    OpenAIRE

    Bounkeua, Viengngeun

    2010-01-01

    This work explored sexual development of the lethal human malaria parasite, Plasmodium falciparum. A method to cultivate Plasmodium falciparum sexual stage parasites in vitro was optimized and increased ookinete yield > 20-fold over previous reports. This method was adapted for Plasmodium vivax, a neglected human malaria parasite, and found to generate Plasmodium vivax ookinetes. The method was essential for investigation of the role of a novel aspartic protease, Plasmepsin X, in sexual stage...

  13. Natural microbe-mediated refractoriness to Plasmodium infection in Anopheles gambiae

    OpenAIRE

    Cirimotich, Chris M; Dong, Yuemei; Clayton, April M.; Sandiford, Simone L.; Jayme A Souza-Neto; Mulenga, Musapa; Dimopoulos, George

    2011-01-01

    Malaria parasite transmission depends on the successful transition of Plasmodium through discrete developmental stages in the lumen of the mosquito midgut. Like the human intestinal tract, the mosquito midgut contains a diverse microbial flora, which may compromise the ability of Plasmodium to establish infection. We have identified an Enterobacter bacterium isolated from wild mosquito populations in Zambia that renders the mosquito 99% resistant to infection with the human malaria parasite P...

  14. Single Sensillum Recordings in the Insects Drosophila melanogaster and Anopheles gambiae

    OpenAIRE

    Pellegrino, Maurizio; Nakagawa, Takao; Vosshall, Leslie B.

    2010-01-01

    The sense of smell is essential for insects to find foods, mates, predators, and oviposition sites3. Insect olfactory sensory neurons (OSNs) are enclosed in sensory hairs called sensilla, which cover the surface of olfactory organs. The surface of each sensillum is covered with tiny pores, through which odorants pass and dissolve in a fluid called sensillum lymph, which bathes the sensory dendrites of the OSNs housed in a given sensillum. The OSN dendrites express odorant receptor (OR) protei...

  15. Hemocyte differentiation mediates the mosquito late-phase immune response against Plasmodium in Anopheles gambiae.

    Science.gov (United States)

    Smith, Ryan C; Barillas-Mury, Carolina; Jacobs-Lorena, Marcelo

    2015-06-30

    Plasmodium parasites must complete development in the mosquito vector for transmission to occur. The mosquito innate immune response is remarkably efficient in limiting parasite numbers. Previous work has identified a LPS-induced TNFα transcription factor (LITAF)-like transcription factor, LITAF-like 3 (LL3), which significantly influences parasite numbers. Here, we demonstrate that LL3 does not influence invasion of the mosquito midgut epithelium or ookinete-to-oocyst differentiation but mediates a late-phase immune response that decreases oocyst survival. LL3 expression in the midgut and hemocytes is activated by ookinete midgut invasion and is independent of the mosquito microbiota, suggesting that LL3 may be a component of a wound-healing response. LL3 silencing abrogates the ability of mosquito hemocytes to differentiate and respond to parasite infection, implicating hemocytes as critical modulators of the late-phase immune response. PMID:26080400

  16. Hemocyte differentiation mediates the mosquito late-phase immune response against Plasmodium in Anopheles gambiae

    OpenAIRE

    Smith, Ryan C.; Barillas-Mury, Carolina; Jacobs-Lorena, Marcelo

    2015-01-01

    The innate immune response is a major determinant of malaria parasite success in its mosquito host. Previous experiments have implicated LPS-induced TNFα transcription factor (LITAF)-like 3 (LL3) as an integral component of the mosquito immune response to the malaria parasite. This study reports that LL3 influences oocyst survival and demonstrates its role in mosquito blood cell (hemocyte) differentiation in response to parasite infection. Integrating previous data, we provide evidence that h...

  17. Hemocyte differentiation mediates the mosquito late-phase immune response against Plasmodium in Anopheles gambiae

    Science.gov (United States)

    Smith, Ryan C.; Barillas-Mury, Carolina; Jacobs-Lorena, Marcelo

    2015-01-01

    Plasmodium parasites must complete development in the mosquito vector for transmission to occur. The mosquito innate immune response is remarkably efficient in limiting parasite numbers. Previous work has identified a LPS-induced TNFα transcription factor (LITAF)-like transcription factor, LITAF-like 3 (LL3), which significantly influences parasite numbers. Here, we demonstrate that LL3 does not influence invasion of the mosquito midgut epithelium or ookinete-to-oocyst differentiation but mediates a late-phase immune response that decreases oocyst survival. LL3 expression in the midgut and hemocytes is activated by ookinete midgut invasion and is independent of the mosquito microbiota, suggesting that LL3 may be a component of a wound-healing response. LL3 silencing abrogates the ability of mosquito hemocytes to differentiate and respond to parasite infection, implicating hemocytes as critical modulators of the late-phase immune response. PMID:26080400

  18. The bionomics of the malaria mosquito Anopheles gambiae sensu lato in Southeast Tanzania.

    NARCIS (Netherlands)

    Lyimo, E.O.K.

    1993-01-01

    Size of adult mosquitoes is known to affect both population dynamics as well as disease transmission. Studies devoted to this topic have given different results for different species. For example in some mosquito species, large size was found to be associated with high fecundity and longer survival

  19. The bionomics of the malaria mosquito Anopheles gambiae sensu lato in Southeast Tanzania.

    OpenAIRE

    Lyimo, E.O.K.

    1993-01-01

    Size of adult mosquitoes is known to affect both population dynamics as well as disease transmission. Studies devoted to this topic have given different results for different species. For example in some mosquito species, large size was found to be associated with high fecundity and longer survival (Steinwascher, 1982; Nasci, 1986a; 1986b; 1987) but in others large size did not result in longer survival (Walker et al ., 1987; Landry et al ., 1988; Pumpuni & Walker, 1989). Similar data were fo...

  20. Status of insecticide susceptibility in Anopheles arabiensis from Mwea rice irrigation scheme, Central Kenya

    Directory of Open Access Journals (Sweden)

    Vulule John M

    2006-06-01

    Full Text Available Abstract Background Control of the Anopheline mosquito vectors of malaria by use of insecticides has been shown to impact on both morbidity and mortality due to this disease. Evidence of insecticide resistance in different settings necessitates surveillance studies to allow prompt detection of resistance should it arise and thus enable its management. Possible resistance by Anopheles arabiensis mosquitoes from Mwea rice irrigation scheme in Central Kenya to insecticides in the four classes of insecticides approved by WHO for indoor residual spraying was investigated. Methods Susceptibility to DDT (an organochlorine, fenitrothion (an organophosphate, bendiocarb (a carbamate, lambdacyhalothrin and permethrin (both pyrethroids was tested using standard WHO diagnostic bioassay kits. Bioassays were performed on non-blood fed mosquitoes one- to three-day old. Knockdown was recorded every 10 min and mortality 24 h post-exposure was noted. Results Mortality 24 h post-exposure was 100% for all insecticides except for lambdacyhalothrin, which averaged 99.46%. Knockdown rates at 10 min intervals were not significantly different between the Mwea population and the susceptible KISUMU strain of Anopheles gambiae sensu stricto control. The KDT50 and KDT95 values for the Mwea population were either lower than those for the control or higher by factors of no more than 2 for most comparisons and compared well with those of An. gambiae sensu lato categorized as susceptible in other studies. Conclusion These results suggest that the Mwea population of An. arabiensis is susceptible to all the insecticides tested. This implies that vector control measures employing any of these insecticides would not be hampered by resistance.

  1. A new mtDNA COI gene lineage closely related to Anopheles janconnae of the Albitarsis complex in the Caribbean region of Colombia

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    Lina A Gutiérrez

    2010-12-01

    Full Text Available An understanding of the taxonomic status and vector distribution of anophelines is crucial in controlling malaria. Previous phylogenetic analyses have supported the description of six species of the Neotropical malaria vector Anopheles (Nyssorhynchus albitarsis s.l. (Diptera: Culicidae: An. albitarsis, Anopheles deaneorum, Anopheles marajoara, Anopheles oryzalimnetes, Anopheles janconnae and An. albitarsis F. To evaluate the taxonomic status of An. albitarsis s.l. mosquitoes collected in various localities in the Colombian Caribbean region, specimens were analyzed using the complete mitochondrial DNA cytochrome oxidase I (COI gene, the ribosomal DNA (rDNA internal transcribed spacer 2 (ITS2 region and partial nuclear DNA white gene sequences. Phylogenetic analyses of the COI gene sequences detected a new lineage closely related to An. janconnae in the Caribbean region of Colombia and determined its position relative to the other members of the complex. However, the ITS2 and white gene sequences lacked sufficient resolution to support a new lineage closely related to An. janconnae or the An. janconnae clade. The possible involvement of this new lineage in malaria transmission in Colombia remains unknown, but its phylogenetic closeness to An. janconnae, which has been implicated in local malaria transmission in Brazil, is intriguing.

  2. Life-table analysis of Anopheles malaria vectors: generational mortality as tool in mosquito vector abundance and control studies

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    Godwin Ray Anugboba Okogun

    2005-06-01

    Full Text Available Background & objectives: Vector control will for sometime remain a primary weapon in the waragainst vector borne diseases. Malaria is of paramount importance in this with its associated highmorbidity and mortality especially in sub-Saharan Africa. This study on generational mortality associatedfactors in Anopheles mosquitoes life-table analysis was designed to investigate the fecundity,levels of mortality and mortality associated factors at the aquatic stages of anopheline malaria vectors.Methods: Mortality associated factors were investigated at the eggs, I and II instar larval, III and IVinstar larval and pupal stages of two anopheline species— Anopheles pseudopunctipennis (Theobaldand An. gambiae life-cycles in screen cages. Adult male and female mosquitoes were membrane filterfedand algae in culture medium formed the bulk of food substances for the larval stage. Environmentaltemperature of culture media, pH and some associated physio-chemical factors were also determined.Results: Results showed significant mortality rates at various aquatic stages. Infertility, cannibalismand environmental factors were the major factors responsible for mortality at the egg, larval and pupalstages respectively.Interpretation & conclusion: The aquatic stages of Anopheles mosquito mortality factor K and themortality factors at the various stages investigated k1, k2, k3 and k4 are discussed. Our recommendationsinclude further studies on the possible genetic modification of predacious An. pseudopunctipennislarvae and/or its modification for the production of sterile/infertile eggs as possible alternativesin the reduction and control of anopheline malaria burden.

  3. A natural Anopheles-associated Penicillium chrysogenum enhances mosquito susceptibility to Plasmodium infection

    Science.gov (United States)

    Angleró-Rodríguez, Yesseinia I.; Blumberg, Benjamin J.; Dong, Yuemei; Sandiford, Simone L.; Pike, Andrew; Clayton, April M.; Dimopoulos, George

    2016-01-01

    Whereas studies have extensively examined the ability of bacteria to influence Plasmodium infection in the mosquito, the tripartite interactions between non-entomopathogenic fungi, mosquitoes, and Plasmodium parasites remain largely uncharacterized. Here we report the isolation of a common mosquito-associated ascomycete fungus, Penicillium chrysogenum, from the midgut of field-caught Anopheles mosquitoes. Although the presence of Pe. chrysogenum in the Anopheles gambiae midgut does not affect mosquito survival, it renders the mosquito significantly more susceptible to Plasmodium infection through a secreted heat-stable factor. We further provide evidence that the mechanism of the fungus-mediated modulation of mosquito susceptibility to Plasmodium involves an upregulation of the insect’s ornithine decarboxylase gene, which sequesters arginine for polyamine biosynthesis. Arginine plays an important role in the mosquito’s anti-Plasmodium defense as a substrate of nitric oxide production, and its availability therefore has a direct impact on the mosquito’s susceptibility to the parasite. While this type of immunomodulatory mechanism has already been demonstrated in other host-pathogen interaction systems, this is the first report of a mosquito-associated fungus that can suppress the mosquito’s innate immune system in a way that would favor Plasmodium infection and possibly malaria transmission. PMID:27678168

  4. Recent reduction in the water level of Lake Victoria has created more habitats for Anopheles funestus

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    Futami Kyoko

    2008-07-01

    Full Text Available Abstract Background The water level of Lake Victoria has fallen more than 1.5 m since 1998, revealing a narrow strip of land along the shore. This study determined whether the recent drop in the water level has created additional breeding grounds for malaria vectors. Methods The recent and past shorelines were estimated using landmarks and a satellite image. The locations of breeding habitats were recorded using a GPS unit during the high and low lake water periods. GIS was used to determine whether the breeding habitats were located on newly emerged land between the new and old shorelines. Results Over half of the breeding habitats existed on newly emerged land. Fewer habitats for the Anopheles gambiae complex were found during the low water level period compared to the high water period. However, more habitats for Anopheles funestus were found during the high water level period, and they were all located on the newly emerged land. Conclusion The recent reduction in water level of Lake Victoria has increased the amount of available habitat for A. funestus. The results suggest that the water drop has substantially affected the population of this malaria vector in the Lake Victoria basin, particularly because the lake has a long shoreline that may harbour many new breeding habitats.

  5. Mosquito repellent action of Blumea lacera (Asteraceae against Anopheles stephensi and Culex quinquefasciatus.

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    S.P. Singh

    2014-03-01

    Full Text Available Petroleum ether extract of Blumea lacera was screened under laboratory conditions for repellent activity against mosquito vector Anopheles stephensi Liston and Culex quinquefasciatus Say (Diptera: Culicidae. The repellent activity of Blumea lacera extract was tested against mosquitoes in comparison with the DEET, which was used as a positive control. Results obtained from the laboratory experiment showed that the extract was effective against mosquito vectors even at a low dose. A direct relationship was observed with concentrations of Blumea lacera extract and the repellent activity. Percent repellency obtained at 6% concentration of theextract against An. stephensi and Cx. quinquefasciatus were 97and 98% at 0 hour and 78.8 and 76.2% after 6 hrs. DEET-2% however showed 100% repellency against An. stephensi and against Cx. quinquefasciatus up to 4 hours and 1 hour, respectively. These results show that Blumea lacera extract has the potential as an effective mosquito repellent.

  6. Genetic Study of Propoxur Resistance—A Carbamate Insecticide in the Malaria Mosquito, Anopheles stephensi Liston

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    D. Sanil

    2010-01-01

    Full Text Available Anopheles stephensi Liston (Diptera: Culicidae is the urban vector of malaria in the Indian subcontinent and several countries of the Middle East. The genetics of propoxur resistance (pr in An. stephensi larvae was studied to determine its mode of inheritance. A diagnostic dose of 0.01 mg/L as recommended by WHO was used to establish homozygous resistant and susceptible strains. Reciprocal crosses between the resistant and susceptible strains showed an F1 generation of incomplete dominance. The progenies of backcrosses to susceptible parents were in 1 : 1 ratio of the same phenotypes as the parents and hybrids involved. The dosage mortality (d-m lines were constructed for each one of the crosses, and the degree of dominance was calculated. It is concluded that propoxur resistance in An. stephensi larvae is due to monofactorial inheritance with incomplete dominance and is autosomal in nature.

  7. Comparative analyses reveal discrepancies among results of commonly used methods for Anopheles gambiaemolecular form identification

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    Pinto João

    2011-08-01

    Full Text Available Abstract Background Anopheles gambiae M and S molecular forms, the major malaria vectors in the Afro-tropical region, are ongoing a process of ecological diversification and adaptive lineage splitting, which is affecting malaria transmission and vector control strategies in West Africa. These two incipient species are defined on the basis of single nucleotide differences in the IGS and ITS regions of multicopy rDNA located on the X-chromosome. A number of PCR and PCR-RFLP approaches based on form-specific SNPs in the IGS region are used for M and S identification. Moreover, a PCR-method to detect the M-specific insertion of a short interspersed transposable element (SINE200 has recently been introduced as an alternative identification approach. However, a large-scale comparative analysis of four widely used PCR or PCR-RFLP genotyping methods for M and S identification was never carried out to evaluate whether they could be used interchangeably, as commonly assumed. Results The genotyping of more than 400 A. gambiae specimens from nine African countries, and the sequencing of the IGS-amplicon of 115 of them, highlighted discrepancies among results obtained by the different approaches due to different kinds of biases, which may result in an overestimation of MS putative hybrids, as follows: i incorrect match of M and S specific primers used in the allele specific-PCR approach; ii presence of polymorphisms in the recognition sequence of restriction enzymes used in the PCR-RFLP approaches; iii incomplete cleavage during the restriction reactions; iv presence of different copy numbers of M and S-specific IGS-arrays in single individuals in areas of secondary contact between the two forms. Conclusions The results reveal that the PCR and PCR-RFLP approaches most commonly utilized to identify A. gambiae M and S forms are not fully interchangeable as usually assumed, and highlight limits of the actual definition of the two molecular forms, which might

  8. Plasmodium falciparum infection rates for some Anopheles spp. from Guinea-Bissau, West Africa [v1; ref status: indexed, http://f1000r.es/4in

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    Michelle R. Sanford

    2014-10-01

    Full Text Available Presence of Plasmodium falciparum circumsporozoite protein (CSP was detected by enzyme linked immunosorbent assay (ELISA in a sample of Anopheles gambiae s.s., A. melas and A. pharoensis collected in Guinea-Bissau during October and November 2009. The percentage of P. falciparum infected samples (10.2% overall was comparable to earlier studies from other sites in Guinea-Bissau (9.6-12.4%. The majority of the specimens collected were identified as A. gambiae which had an individual infection rate of 12.6 % across collection sites. A small number of specimens of A. coluzzii, A. coluzzii x A. gambiae hybrids, A. melas and A. pharoensis were collected and had infection rates of 4.3%, 4.1%, 11.1% and 33.3% respectively. Despite being present in low numbers in indoor collections, the exophilic feeding behaviors of A. melas (N=18 and A. pharoensis (N=6 and high infection rates observed in this survey suggest falciparum-malaria transmission potential outside of the protection of bed nets.

  9. An Evolution-Based Screen for Genetic Differentiation between Anopheles Sister Taxa Enriches for Detection of Functional Immune Factors.

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    Christian Mitri

    2015-12-01

    Full Text Available Nucleotide variation patterns across species are shaped by the processes of natural selection, including exposure to environmental pathogens. We examined patterns of genetic variation in two sister species, Anopheles gambiae and Anopheles coluzzii, both efficient natural vectors of human malaria in West Africa. We used the differentiation signature displayed by a known coordinate selective sweep of immune genes APL1 and TEP1 in A. coluzzii to design a population genetic screen trained on the sweep, classified a panel of 26 potential immune genes for concordance with the signature, and functionally tested their immune phenotypes. The screen results were strongly predictive for genes with protective immune phenotypes: genes meeting the screen criteria were significantly more likely to display a functional phenotype against malaria infection than genes not meeting the criteria (p = 0.0005. Thus, an evolution-based screen can efficiently prioritize candidate genes for labor-intensive downstream functional testing, and safely allow the elimination of genes not meeting the screen criteria. The suite of immune genes with characteristics similar to the APL1-TEP1 selective sweep appears to be more widespread in the A. coluzzii genome than previously recognized. The immune gene differentiation may be a consequence of adaptation of A. coluzzii to new pathogens encountered in its niche expansion during the separation from A. gambiae, although the role, if any of natural selection by Plasmodium is unknown. Application of the screen allowed identification of new functional immune factors, and assignment of new functions to known factors. We describe biochemical binding interactions between immune proteins that underlie functional activity for malaria infection, which highlights the interplay between pathogen specificity and the structure of immune complexes. We also find that most malaria-protective immune factors display phenotypes for either human or rodent

  10. An Evolution-Based Screen for Genetic Differentiation between Anopheles Sister Taxa Enriches for Detection of Functional Immune Factors.

    Science.gov (United States)

    Mitri, Christian; Bischoff, Emmanuel; Takashima, Eizo; Williams, Marni; Eiglmeier, Karin; Pain, Adrien; Guelbeogo, Wamdaogo M; Gneme, Awa; Brito-Fravallo, Emma; Holm, Inge; Lavazec, Catherine; Sagnon, N'Fale; Baxter, Richard H; Riehle, Michelle M; Vernick, Kenneth D

    2015-12-01

    Nucleotide variation patterns across species are shaped by the processes of natural selection, including exposure to environmental pathogens. We examined patterns of genetic variation in two sister