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Sample records for anopheles gambiae mosquitoes

  1. Plasmodium activates the innate immune response of Anopheles gambiae mosquitoes.

    OpenAIRE

    Richman, A M; Dimopoulos, G; Seeley, D; Kafatos, F C

    1997-01-01

    Innate immune-related gene expression in the major disease vector mosquito Anopheles gambiae has been analyzed following infection by the malaria parasite, Plasmodium berghei. Substantially increased levels of mRNAs encoding the antibacterial peptide defensin and a putative Gram-negative bacteria-binding protein (GNBP) are observed 20-30 h after ingestion of an infected blood-meal, at a time which indicates that this induction is a response to parasite invasion of the midgut epithelium. The i...

  2. 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

  3. A maleness gene in the malaria mosquito Anopheles gambiae.

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    Krzywinska, Elzbieta; Dennison, Nathan J; Lycett, Gareth J; Krzywinski, Jaroslaw

    2016-07-01

    The molecular pathways controlling gender are highly variable and have been identified in only a few nonmammalian model species. In many insects, maleness is conferred by a Y chromosome-linked M factor of unknown nature. We have isolated and characterized a gene, Yob, for the M factor in the malaria mosquito Anopheles gambiae Yob, activated at the beginning of zygotic transcription and expressed throughout a male's life, controls male-specific splicing of the doublesex gene. Silencing embryonic Yob expression is male-lethal, whereas ectopic embryonic delivery of Yob transcripts yields male-only broods. This female-killing property may be an invaluable tool for creation of conditional male-only transgenic Anopheles strains for malaria control programs.

  4. Insecticide susceptibility of Anopheles coluzzii and Anopheles gambiae mosquitoes in Ibadan, Southwest Nigeria.

    Science.gov (United States)

    Okorie, P N; Ademowo, O G; Irving, H; Kelly-Hope, L A; Wondji, C S

    2015-03-01

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

  5. 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...

  6. Biolistic techniques for transfection of mosquito embryos (Anopheles gambiae).

    Science.gov (United States)

    Mialhe, E; Miller, L H

    1994-05-01

    To compensate for the extremely low rates of transformation by DNA microinjection into mosquito embryos of Anopheles gambiae, biolistic techniques were evaluated for introduction of DNA into large numbers of mosquito embryos. Biolistic experiments were first performed with a commercially available instrument intended for this purpose, according to the recommended procedure. The amount of DNA delivered was measured by the expression of luciferase under the control of the Drosophila heat shock protein (hsp) 70 promoter. Despite attempts to optimize biolistic parameters, the level of luciferase activity was low and highly variable. Two other methods of biolistic delivery of DNA-coated particles in aqueous suspension were then evaluated. One method used the gas explosion of the commercially available instrument (mentioned above) to drive an aqueous suspension of DNA-coated particles at high pressure. This method reproducibly increased the level of expression about 100-fold without greatly reducing embryo viability. Another method, which was recently described for plant transfection, uses lower pressure to deliver the aqueous suspension of DNA-coated particles. The level of expression of luciferase and the survival of embryos were equivalent to that obtained with the instrument modified for aqueous delivery of particles. Thus, both aqueous methods offer the advantages of reproducibly delivering more DNA to the embryos. Moreover, these methods could be suitable for delivering DNA mixed with proteins, such as restriction endonucleases and integrases, that may be destroyed by ethanol precipitation used in the standard PDS-1000/He method.

  7. Factors influencing infection and transmission of Anopheles gambiae densovirus (AgDNV) in mosquitoes.

    Science.gov (United States)

    Barik, Tapan K; Suzuki, Yasutsugu; Rasgon, Jason L

    2016-01-01

    Anopheles gambiae densovirus (AgDNV) is a potential microbial agent for paratransgenesis and gene transduction in An. gambiae, the major vector of human malaria in sub-Saharan Africa. Understanding the interaction between AgDNV and An. gambiae is critical for using AgDNV in a basic and applied manner for Anopheles gene manipulation. Here, we tested the effects of mosquito age, sex, blood feeding status, and potential for horizontal transmission using an enhanced green fluorescent protein (EGFP) reporter AgDNV system. Neither mosquito age at infection nor feeding regime affected viral titers. Female mosquitoes were more permissive to viral infection than males. Despite low viral titers, infected males were able to venereally transmit virus to females during mating, where the virus was localized with the transferred sperm in the spermathecae. These findings will be useful for designing AgDNV-based strategies to manipulate Anopheles gambiae.

  8. Factors influencing infection and transmission of Anopheles gambiae densovirus (AgDNV in mosquitoes

    Directory of Open Access Journals (Sweden)

    Tapan K. Barik

    2016-11-01

    Full Text Available Anopheles gambiae densovirus (AgDNV is a potential microbial agent for paratransgenesis and gene transduction in An. gambiae, the major vector of human malaria in sub-Saharan Africa. Understanding the interaction between AgDNV and An. gambiae is critical for using AgDNV in a basic and applied manner for Anopheles gene manipulation. Here, we tested the effects of mosquito age, sex, blood feeding status, and potential for horizontal transmission using an enhanced green fluorescent protein (EGFP reporter AgDNV system. Neither mosquito age at infection nor feeding regime affected viral titers. Female mosquitoes were more permissive to viral infection than males. Despite low viral titers, infected males were able to venereally transmit virus to females during mating, where the virus was localized with the transferred sperm in the spermathecae. These findings will be useful for designing AgDNV-based strategies to manipulate Anopheles gambiae.

  9. 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...

  10. Plasmodium activates the innate immune response of Anopheles gambiae mosquitoes.

    Science.gov (United States)

    Richman, A M; Dimopoulos, G; Seeley, D; Kafatos, F C

    1997-01-01

    Innate immune-related gene expression in the major disease vector mosquito Anopheles gambiae has been analyzed following infection by the malaria parasite, Plasmodium berghei. Substantially increased levels of mRNAs encoding the antibacterial peptide defensin and a putative Gram-negative bacteria-binding protein (GNBP) are observed 20-30 h after ingestion of an infected blood-meal, at a time which indicates that this induction is a response to parasite invasion of the midgut epithelium. The induction is dependent upon the ingestion of infective, sexual-stage parasites, and is not due to opportunistic co-penetration of resident gut micro-organisms into the hemocoel. The response is activated following infection both locally (in the midgut) and systemically (in remaining tissues, presumably fat body and/or hemocytes). The observation that Plasmodium can trigger a molecularly defined immune response in the vector constitutes an important advance in our understanding of parasite-vector interactions that are potentially involved in malaria transmission, and extends knowledge of the innate immune system of insects to encompass responses to protozoan parasites. PMID:9321391

  11. 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.

  12. 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 previo...

  13. 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

  14. 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

  15. Blood feeding induces hemocyte proliferation and activation in the African malaria mosquito, Anopheles gambiae Giles

    OpenAIRE

    Bryant, William B.; Michel, Kristin

    2014-01-01

    Malaria is a global public health problem, especially in sub-Saharan Africa, where the mosquito Anopheles gambiae Giles serves as the major vector for the protozoan Plasmodium falciparum Welch. One determinant of malaria vector competence is the mosquito's immune system. Hemocytes are a critical component as they produce soluble immune factors that either support or prevent malaria parasite development. However, despite their importance in vector competence, understanding of their basic biolo...

  16. A viral over-expression system for the major malaria mosquito Anopheles gambiae.

    Science.gov (United States)

    Suzuki, Yasutsugu; Niu, Guodong; Hughes, Grant L; Rasgon, Jason L

    2014-05-30

    Understanding pathogen/mosquito interactions is essential for developing novel strategies to control mosquito-borne diseases. Technical advances in reverse-genetics, such as RNA interference (RNAi), have facilitated elucidation of components of the mosquito immune system that are antagonistic to pathogen development, and host proteins essential for parasite development. Forward genetic approaches, however, are limited to generation of transgenic insects, and while powerful, mosquito transgenesis is a resource- and time-intensive technique that is not broadly available to most laboratories. The ability to easily "over-express" genes would enhance molecular studies in vector biology and expedite elucidation of pathogen-refractory genes without the need to make transgenic insects. We developed and characterized an efficient Anopheles gambiae densovirus (AgDNV) over-expression system for the major malaria vector Anopheles gambiae. High-levels of gene expression were detected at 3 days post-infection and increased over time, suggesting this is an effective system for gene induction. Strong expression was observed in the fat body and ovaries. We validated multiple short promoters for gene induction studies. Finally, we developed a polycistronic system to simultaneously express multiple genes of interest. This AgDNV-based toolset allows for consistent transduction of genes of interest and will be a powerful molecular tool for research in Anopheles gambiae mosquitoes.

  17. Genomic and evolutionary analyses of Tango transposons in Aedes aegypti, Anopheles gambiae and other mosquito species.

    Science.gov (United States)

    Coy, M R; Tu, Z

    2007-08-01

    Tango is a transposon of the Tc1 family and was originally discovered in the African malaria mosquito, Anopheles gambiae. Here we report a systematic analysis of the genome sequence of the yellow fever mosquito, Aedes aegypti, which uncovered three distinct Tango transposons. We name the only An. gambiae Tango transposon AgTango1 and the three Ae. aegypti Tango elements AeTango1-3. Like AgTango1, AeTango1 and AeTango2 elements both have members that retain characteristics of autonomous elements such as intact open reading frames and terminal inverted repeats (TIRs). AeTango3 is a degenerate transposon with no full-length members. All full-length Tango transposons contain subterminal direct repeats within their TIRs. AgTango1 and AeTango1-3 form a single clade among other Tc1 transposons. Within this clade, AgTango1 and AeTango1 are closely related and share approximately 80% identity at the amino acid level, which exceeds the level of similarity of the majority of host genes in the two species. A survey of Tango in other mosquito species was carried out using degenerate PCR. Tango was isolated and sequenced in all members of the An. gambiae species complex, Aedes albopictus and Ochlerotatus atropalpus. Oc. atropalpus contains a rich diversity of Tango elements, while Tango elements in Ae. albopictus and the An. gambiae species complex all belong to Tango1. No Tango was detected in Culex pipiens quinquefasciatus, Anopheles stephensi, Anopheles dirus, Anopheles farauti or Anopheles albimanus using degenerate PCR. Bioinformatic searches of the Cx. p. quinquefasciatus (~10 x coverage) and An. stephensi (0.33 x coverage) databases also failed to uncover any Tango elements. Although other evolutionary scenarios cannot be ruled out, there are indications that Tango1 underwent horizontal transfer among divergent mosquito species.

  18. 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.

  19. Thorsellia anophelis is the dominant bacterium in a Kenyan population of adult Anopheles gambiae mosquitoes.

    Science.gov (United States)

    Briones, Aurelio M; Shililu, Josephat; Githure, John; Novak, Robert; Raskin, Lutgarde

    2008-01-01

    Anopheles gambiae mosquitoes are not known to harbor endosymbiotic bacteria. Here we show, using nucleic acid-based methods, that 16S rRNA gene sequences specific to a recently described mosquito midgut bacterium, Thorsellia anophelis, is predominant in the midgut of adult An. gambiae s.l. mosquitoes captured in residences in central Kenya, and also occurs in the aquatic rice paddy environment nearby. PCR consistently detected T. anophelis in the surface microlayer of rice paddies, which is also consistent with the surface-feeding behavior of A. gambiae s.l. larvae. Phylogenetic analysis of cloned environmental 16S rRNA genes identified four major Thorsellia lineages, which are closely affiliated to an insect endosymbiont of the genus Arsenophonus. Physiological characterizations support the hypothesis that T. anophelis is well adapted to the female anopheline midgut by utilizing blood and tolerating the alkaline conditions in this environment. The results suggest that aquatically derived bacteria such as T. anophelis can persist through mosquito metamorphosis and become well-established in the adult mosquito midgut.

  20. 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

  1. Transgenic Expression of the Anti-parasitic Factor TEP1 in the Malaria Mosquito Anopheles gambiae

    Science.gov (United States)

    Hopp, Ann-Katrin; Saenger, Mélanie; Soichot, Julien; Scholze, Heidi; Boch, Jens; Blandin, Stéphanie A.; Marois, Eric

    2017-01-01

    Mosquitoes genetically engineered to be resistant to Plasmodium parasites represent a promising novel approach in the fight against malaria. The insect immune system itself is a source of anti-parasitic genes potentially exploitable for transgenic designs. The Anopheles gambiae thioester containing protein 1 (TEP1) is a potent anti-parasitic protein. TEP1 is secreted and circulates in the mosquito hemolymph, where its activated cleaved form binds and eliminates malaria parasites. Here we investigated whether TEP1 can be used to create malaria resistant mosquitoes. Using a GFP reporter transgene, we determined that the fat body is the main site of TEP1 expression. We generated transgenic mosquitoes that express TEP1r, a potent refractory allele of TEP1, in the fat body and examined the activity of the transgenic protein in wild-type or TEP1 mutant genetic backgrounds. Transgenic TEP1r rescued loss-of-function mutations, but did not increase parasite resistance in the presence of a wild-type susceptible allele. Consistent with previous reports, TEP1 protein expressed from the transgene in the fat body was taken up by hemocytes upon a challenge with injected bacteria. Furthermore, although maturation of transgenic TEP1 into the cleaved form was impaired in one of the TEP1 mutant lines, it was still sufficient to reduce parasite numbers and induce parasite melanization. We also report here the first use of Transcription Activator Like Effectors (TALEs) in Anopheles gambiae to stimulate expression of endogenous TEP1. We found that artificial elevation of TEP1 expression remains moderate in vivo and that enhancement of endogenous TEP1 expression did not result in increased resistance to Plasmodium. Taken together, our results reveal the difficulty of artificially influencing TEP1-mediated Plasmodium resistance, and contribute to further our understanding of the molecular mechanisms underlying mosquito resistance to Plasmodium parasites. PMID:28095489

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

    Science.gov (United States)

    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. Dynamic gut microbiome across life history of the malaria mosquito Anopheles gambiae in Kenya.

    Directory of Open Access Journals (Sweden)

    Ying Wang

    Full Text Available The mosquito gut represents an ecosystem that accommodates a complex, intimately associated microbiome. It is increasingly clear that the gut microbiome influences a wide variety of host traits, such as fitness and immunity. Understanding the microbial community structure and its dynamics across mosquito life is a prerequisite for comprehending the symbiotic relationship between the mosquito and its gut microbial residents. Here we characterized gut bacterial communities across larvae, pupae and adults of Anopheles gambiae reared in semi-natural habitats in Kenya by pyrosequencing bacterial 16S rRNA fragments. Immatures and adults showed distinctive gut community structures. Photosynthetic Cyanobacteria were predominant in the larval and pupal guts while Proteobacteria and Bacteroidetes dominated the adult guts, with core taxa of Enterobacteriaceae and Flavobacteriaceae. At the adult stage, diet regime (sugar meal and blood meal significantly affects the microbial structure. Intriguingly, blood meals drastically reduced the community diversity and favored enteric bacteria. Comparative genomic analysis revealed that the enriched enteric bacteria possess large genetic redox capacity of coping with oxidative and nitrosative stresses that are associated with the catabolism of blood meal, suggesting a beneficial role in maintaining gut redox homeostasis. Interestingly, gut community structure was similar in the adult stage between the field and laboratory mosquitoes, indicating that mosquito gut is a selective eco-environment for its microbiome. This comprehensive gut metatgenomic profile suggests a concerted symbiotic genetic association between gut inhabitants and host.

  4. 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

  5. Aquaporin water channel AgAQP1 in the malaria vector mosquito Anopheles gambiae during blood feeding and humidity adaptation.

    Science.gov (United States)

    Liu, Kun; Tsujimoto, Hitoshi; Cha, Sung-Jae; Agre, Peter; Rasgon, Jason L

    2011-04-12

    Altered patterns of malaria endemicity reflect, in part, changes in feeding behavior and climate adaptation of mosquito vectors. Aquaporin (AQP) water channels are found throughout nature and confer high-capacity water flow through cell membranes. The genome of the major malaria vector mosquito Anopheles gambiae contains at least seven putative AQP sequences. Anticipating that transmembrane water movements are important during the life cycle of A. gambiae, we identified and characterized the A. gambiae aquaporin 1 (AgAQP1) protein that is homologous to AQPs known in humans, Drosophila, and sap-sucking insects. When expressed in Xenopus laevis oocytes, AgAQP1 transports water but not glycerol. Similar to mammalian AQPs, water permeation of AgAQP1 is inhibited by HgCl(2) and tetraethylammonium, with Tyr185 conferring tetraethylammonium sensitivity. AgAQP1 is more highly expressed in adult female A. gambiae mosquitoes than in males. Expression is high in gut, ovaries, and Malpighian tubules where immunofluorescence microscopy reveals that AgAQP1 resides in stellate cells but not principal cells. AgAQP1 expression is up-regulated in fat body and ovary by blood feeding but not by sugar feeding, and it is reduced by exposure to a dehydrating environment (42% relative humidity). RNA interference reduces AgAQP1 mRNA and protein levels. In a desiccating environment (adaptation of A. gambiae, a major mosquito vector of human malaria in sub-Saharan Africa.

  6. 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.

  7. 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.

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

    Science.gov (United States)

    Olsen, Stine S; Cazzamali, Giuseppe; Williamson, Michael; Grimmelikhuijzen, Cornelis J P; Hauser, Frank

    2007-10-19

    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 5.2x10(-9)M and 6.4x10(-9)M, respectively. However, this receptor could also be activated by the two pyrokinins-1, albeit with lower potency (EC(50): 2-5x10(-8)M). Because Ang-capa-1 and -2 and Ang-PK-1-1 are located on one preprohormone and the other peptides on another prohormone, these results imply a considerable crosstalk between the capa, pyrokinin-1 and pyrokinin-2 systems. Gene structure and phylogenetic tree analyses showed that Ang-Capa-R was the orthologue of the Drosophila capa receptor CG14575, Ang-PK-1-R the orthologue of the Drosophila pyrokinin-1 receptor CG9918, and Ang-PK-2-R the orthologue of the Drosophila pyrokinin-2 receptors CG8784 and CG8795. This is the first report on the functional characterization and crosstalk properties of capa and pyrokinin receptors in mosquitoes.

  9. Human skin emanations in the host-seeking behaviour of the malaria mosquito Anopheles gambiae

    NARCIS (Netherlands)

    Braks, M.A.H.

    1999-01-01

    Malaria is an infectious disease caused by a parasite ( Plasmodium spp.) that is transmitted between human individuals by mosquitoes, belonging to the order of insects, Diptera, family of Culicidae (mosquitoes) and genus of Anopheles (malaria mosquitoes). Mosquitoes feed on humans (and other animals

  10. The Anopheles gambiae odorant binding protein 1 (AgamOBP1 mediates indole recognition in the antennae of female mosquitoes.

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    Harald Biessmann

    Full Text Available Haematophagous insects are frequently carriers of parasitic diseases, including malaria. The mosquito Anopheles gambiae is the major vector of malaria in sub-Saharan Africa and is thus responsible for thousands of deaths daily. Although the role of olfaction in A. gambiae host detection has been demonstrated, little is known about the combinations of ligands and odorant binding proteins (OBPs that can produce specific odor-related responses in vivo. We identified a ligand, indole, for an A. gambiae odorant binding protein, AgamOBP1, modeled the interaction in silico and confirmed the interaction using biochemical assays. RNAi-mediated gene silencing coupled with electrophysiological analyses confirmed that AgamOBP1 binds indole in A. gambiae and that the antennal receptor cells do not respond to indole in the absence of AgamOBP1. This case represents the first documented instance of a specific A. gambiae OBP-ligand pairing combination, demonstrates the significance of OBPs in odor recognition, and can be expanded to the identification of other ligands for OBPs of Anopheles and other medically important insects.

  11. 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

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

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    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.

  13. Using nylon strips to dispense mosquito attractants for sampling the malaria vector Anopheles gambiae s.s.

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    Okumu, F; Biswaro, L; Mbeleyela, E; Killeen, G F; Mukabana, R; Moore, S J

    2010-03-01

    Synthetic versions of human derived kairomones can be used as baits when trapping host seeking mosquitoes. The effectiveness of these lures depends not only on their attractiveness to the mosquitoes but also on the medium from which they are dispensed. We report on the development and evaluation of nylon strips as a method of dispensing odorants attractive to the malaria vector, Anopheles gambiae s.s. (Giles). When a synthetic blend of attractants was dispensed using this method, significantly more mosquitoes were trapped than when two previous methods, open glass vials or low density polyethylene sachets were used. We conclude that the nylon strips are suitable for dispensing odorants in mosquito trapping operations and can be adopted for use in rural and remote areas. The nylon material required is cheap and widely available and the strips can be prepared without specialized equipment or electricity.

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

    Science.gov (United States)

    Deletre, Emilie; Martin, Thibaud; Campagne, Pascal; Bourguet, Denis; Cadin, Andy; Menut, Chantal; Bonafos, Romain; Chandre, Fabrice

    2013-01-01

    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.

  15. 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

  16. 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.

  17. 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.

  18. 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

  19. Molecular Evolution of Immune Genes in the Malaria Mosquito Anopheles gambiae

    Science.gov (United States)

    Lehmann, Tovi; Hume, Jen C. C.; Licht, Monica; Burns, Christopher S.; Wollenberg, Kurt; Simard, Fred; Ribeiro, Jose' M. C.

    2009-01-01

    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 between sibling

  20. Time-of-day specific changes in metabolic detoxification and insecticide resistance in the malaria mosquito Anopheles gambiae.

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    Balmert, Nathaniel J; Rund, Samuel S C; Ghazi, John P; Zhou, Peng; Duffield, Giles E

    2014-05-01

    Mosquitoes exhibit ∼24 h rhythms in physiology and behavior, regulated by the cooperative action of an endogenous circadian clock and the environmental light:dark cycle. Here, we characterize diel (observed under light:dark conditions) time-of-day changes in metabolic detoxification and resistance to insecticide challenge in Anopheles gambiae mosquitoes. A better understanding of mosquito chronobiology will yield insights into developing novel control strategies for this important disease vector. We have previously identified >2000 rhythmically expressed An. gambiae genes. These include metabolic detoxification enzymes peaking at various times throughout the day. Especially interesting was the identification of rhythmic genes encoding enzymes capable of pyrethroid and/or DDT metabolism (CYP6M2, CYP6P3, CYP6Z1, and GSTE2). We hypothesized that these temporal changes in gene expression would confer time-of-day specific changes in metabolic detoxification and responses to insecticide challenge. An. gambiae mosquitoes (adult female Pimperena and Mali-NIH strains) were tested by gene expression analysis for diel rhythms in key genes associated with insecticidal resistance. Biochemical assays for total GST, esterase, and oxidase enzymatic activities were undertaken on time-specific mosquito head and body protein lysates. To determine for rhythmic susceptibility to insecticides by survivorship, mosquitoes were exposed to DDT or deltamethrin across the diel cycle. We report the occurrence of temporal changes in GST activity in samples extracted from the body and head with a single peak at late-night to dawn, but no rhythms were detected in oxidase or esterase activity. The Pimperena strain was found to be resistant to insecticidal challenge, and subsequent genomic analysis revealed the presence of the resistance-conferring kdr mutation. We observed diel rhythmicity in key insecticide detoxification genes in the Mali-NIH strain, with peak phases as previously reported in

  1. Comparative transcriptome analyses of deltamethrin-resistant and -susceptible Anopheles gambiae mosquitoes from Kenya by RNA-Seq.

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    Mariangela Bonizzoni

    Full Text Available Malaria causes more than 300 million clinical cases and 665,000 deaths each year, and the majority of the mortality and morbidity occurs in sub-Saharan Africa. Due to the lack of effective vaccines and wide-spread resistance to antimalarial drugs, mosquito control is the primary method of malaria prevention and control. Currently, malaria vector control relies on the use of insecticides, primarily pyrethroids. The extensive use of insecticides has imposed strong selection pressures for resistance in the mosquito populations. Consequently, resistance to pyrethroids in Anopheles gambiae, the main malaria vector in sub-Saharan Africa, has become a major obstacle for malaria control. A key element of resistance management is the identification of resistance mechanisms and subsequent development of reliable resistance monitoring tools. Field-derived An. gambiae from Western Kenya were phenotyped as deltamethrin-resistant or -susceptible by the standard WHO tube test, and their expression profile compared by RNA-seq. Based on the current annotation of the An. gambiae genome, a total of 1,093 transcripts were detected as significantly differentially accumulated between deltamethrin-resistant and -susceptible mosquitoes. These transcripts are distributed over the entire genome, with a large number mapping in QTLs previously linked to pyrethorid resistance, and correspond to heat-shock proteins, metabolic and transport functions, signal transduction activities, cytoskeleton and others. The detected differences in transcript accumulation levels between resistant and susceptible mosquitoes reflect transcripts directly or indirectly correlated with pyrethroid resistance. RNA-seq data also were used to perform a de-novo Cufflinks assembly of the An. gambiae genome.

  2. Neuropeptides and Peptide Hormones in Anopheles gambiae

    Science.gov (United States)

    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.

  3. 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

  4. 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

  5. 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

  6. 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.

  7. 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.

  8. 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.

  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. Ovicidal and larvicidal activity against Aedes aegypti and Anopheles gambiae complex mosquitoes of essential oils extracted from three spontaneous plants of Burkina Faso.

    Science.gov (United States)

    Bassolé, I H N; Guelbeogo, W M; Nébié, R; Costantini, C; Sagnon, N; Kabore, Z I; Traoré, S A

    2003-03-01

    Essential oils extracted from dried leaves of three spontaneous plants naturally growing in Burkina Faso, i.e. Cymbopogon proximus, Lippia multiflora and Ocimum canum, exhibited larvicidal activity by the WHO standard protocol against 3rd and 4th instar F1-larvae of field-collected mosquitoes vectors of human disease, namely Aedes aegypti and members of the Anopheles gambiae complex, An. arabiensis and An. gambiae. The median lethal concentration (LC50) for Ae. aegypti and An. gambiae s.l. larvae ranged between 53.5-258.5 ppm and 61.9-301.6 ppm, respectively. The LC90 estimates ranged 74.8-334.8 ppm for Ae. aegypti, and 121.6-582.9 ppm for An. gambiae s.l. Ovicidal activity against eggs of An. gambiae s.l. was also demonstrated. The LC50 values for An. gambiae s.l. eggs ranged between 17.1-188.7 ppm, while LC90 values ranged between 33.5-488 ppm. Lippia multiflora showed the highest activity against An. gambiae s.l. eggs and Ae. aegypti larvae, whereas no difference was found among C. proximus and L. multiflora in their activity against An. gambiae s.l. larvae. Of the three plants, essential oils from O. canum had the lowest activity against both eggs and larvae. Eggs were more susceptible than larvae. Ae. aegypti larvae were more susceptible than larvae of An. gambiae s.l.

  11. Multiple origins of knockdown resistance mutations in the Afrotropical mosquito vector Anopheles gambiae.

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    João Pinto

    Full Text Available How often insecticide resistance mutations arise in natural insect populations is a fundamental question for understanding the evolution of resistance and also for modeling its spread. Moreover, the development of resistance is regarded as a favored model to study the molecular evolution of adaptive traits. In the malaria vector Anopheles gambiae two point mutations (L1014F and L1014S in the voltage-gated sodium channel gene, that confer knockdown resistance (kdr to DDT and pyrethroid insecticides, have been described. In order to determine whether resistance alleles result from single or multiple mutation events, genotyping of the kdr locus and partial sequencing of the upstream intron-1 was performed on a total of 288 A. gambiae S-form collected from 28 localities in 15 countries. Knockdown resistance alleles were found to be widespread in West Africa with co-occurrence of both 1014S and 1014F in West-Central localities. Differences in intron-1 haplotype composition suggest that kdr alleles may have arisen from at least four independent mutation events. Neutrality tests provided evidence for a selective sweep acting on this genomic region, particularly in West Africa. The frequency and distribution of these kdr haplotypes varied geographically, being influenced by an interplay between different mutational occurrences, gene flow and local selection. This has important practical implications for the management and sustainability of malaria vector control programs.

  12. Hemolymph circulation in insect sensory appendages: functional mechanics of antennal accessory pulsatile organs (auxiliary hearts) in the mosquito Anopheles gambiae.

    Science.gov (United States)

    Boppana, Sushma; Hillyer, Julián F

    2014-09-01

    Mosquito antennae provide sensory input that modulates host-seeking, mating and oviposition behaviors. Thus, mosquitoes must ensure the efficient transport of molecules into and out of these appendages. To accomplish this, mosquitoes and other insects have evolved antennal accessory pulsatile organs (APOs) that drive hemolymph into the antennal space. This study characterizes the structural mechanics of hemolymph propulsion throughout the antennae of Anopheles gambiae. Using intravital video imaging, we show that mosquitoes possess paired antennal APOs that are located on each side of the head's dorsal midline. They are situated between the frons and the vertex in an area that is dorsal to the antenna but ventral to the medial-most region of the compound eyes. Antennal APOs contract in synchrony at 1 Hz, which is 45% slower than the heart. By means of histology and intravital imaging, we show that each antennal APO propels hemolymph into the antenna through an antennal vessel that traverses the length of the appendage and has an effective diameter of 1-2 μm. When hemolymph reaches the end of the appendage, it is discharged into the antennal hemocoel and returns to the head. Because a narrow vessel empties into a larger cavity, hemolymph travels up the antenna at 0.2 mm s(-1) but reduces its velocity by 75% as it returns to the head. Finally, treatment of mosquitoes with the anesthetic agent FlyNap (triethylamine) increases both antennal APO and heart contraction rates. In summary, this study presents a comprehensive functional characterization of circulatory physiology in the mosquito antennae.

  13. 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.

  14. 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

  15. Comparative molecular modeling of Anopheles gambiae CYP6Z1, a mosquito P450 capable of metabolizing DDT.

    Science.gov (United States)

    Chiu, Ting-Lan; Wen, Zhimou; Rupasinghe, Sanjeewa G; Schuler, Mary A

    2008-07-01

    One of the challenges faced in malarial control is the acquisition of insecticide resistance that has developed in mosquitoes that are vectors for this disease. Anopheles gambiae, which has been the major mosquito vector of the malaria parasite Plasmodium falciparum in Africa, has over the years developed resistance to insecticides including dieldrin, 1,1-bis(p-chlorophenyl)-2,2,2-trichloroethane (DDT), and pyrethroids. Previous microarray studies using fragments of 230 An. gambiae genes identified five P450 loci, including CYP4C27, CYP4H15, CYP6Z1, CYP6Z2, and CYP12F1, that showed significantly higher expression in the DDT-resistant ZAN/U strain compared with the DDT-susceptible Kisumu strain. To predict whether either of the CYP6Z1 and CYP6Z2 proteins might potentially metabolize DDT, we generated and compared molecular models of these two proteins with and without DDT docked in their catalytic sites. This comparison indicated that, although these two CYP6Z proteins share high sequence identity, their metabolic profiles were likely to differ dramatically from the larger catalytic site of CYP6Z1, potentially involved in DDT metabolism, and the more constrained catalytic site of CYP6Z2, not likely to metabolize DDT. Heterologous expressions of these proteins have corroborated these predictions: only CYP6Z1 is capable of metabolizing DDT. Overlays of these models indicate that slight differences in the backbone of SRS1 and variations of side chains in SRS2 and SRS4 account for the significant differences in their catalytic site volumes and DDT-metabolic capacities. These data identify CYP6Z1 as one important target for inhibitor design aimed at inactivating insecticide-metabolizing P450s in natural populations of this malarial mosquito.

  16. 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

  17. An expression map for Anopheles gambiae

    OpenAIRE

    MacCallum Robert M; Redmond Seth N; Christophides George K

    2011-01-01

    Abstract Background Quantitative transcriptome data for the malaria-transmitting mosquito Anopheles gambiae covers a broad range of biological and experimental conditions, including development, blood feeding and infection. Web-based summaries of differential expression for individual genes with respect to these conditions are a useful tool for the biologist, but they lack the context that a visualisation of all genes with respect to all conditions would give. For most organisms, including A....

  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. 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.

  20. A new robust diagnostic polymerase chain reaction for determining the mating status of female Anopheles gambiae mosquitoes

    NARCIS (Netherlands)

    Ng'habi, K.R.; Horton, A.; Knols, B.G.J.; Lanzaro, G.C.

    2007-01-01

    The principal malaria vector in Africa, Anopheles gambiae, contains two pairs of autosomes and one pair of sex chromosomes. The Y chromosome is only associated with males and other Y chromosome¿specific DNA sequences, which are transferred to women during mating. A reliable tool to determine the mat

  1. 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.

  2. Ectopic expression of a cecropin transgene in the human malaria vector mosquito Anopheles gambiae (Diptera: Culicidae): effects on susceptibility to Plasmodium.

    Science.gov (United States)

    Kim, Won; Koo, Hyeyoung; Richman, Adam M; Seeley, Douglas; Vizioli, Jacopo; Klocko, Andrew D; O'Brochta, David A

    2004-05-01

    Genetically altering the disease vector status of insects using recombinant DNA technologies is being considered as an alternative to eradication efforts. Manipulating the endogenous immune response of mosquitoes such as the temporal and special expression of antimicrobial peptides like cecropin may result in a refractory phenotype. Using transgenic technology a unique pattern of expression of cecropin A (cecA) in Anopheles gambiae was created such that cecA was expressed beginning 24 h after a blood meal in the posterior midgut. Two independent lines of transgenic An. gambiae were created using a piggyBac gene vector containing the An. gambiae cecA cDNA under the regulatory control of the Aedes aegypti carboxypeptidase promoter. Infection with Plasmodium berghei resulted in a 60% reduction in the number of oocysts in transgenic mosquitoes compared with nontransgenic mosquitoes. Manipulating the innate immune system of mosquitoes can negatively affect their capacity to serve as hosts for the development of disease-causing microbes.

  3. Malaria infection of the mosquito Anopheles gambiae activates immune-responsive genes during critical transition stages of the parasite life cycle.

    OpenAIRE

    Dimopoulos, G; Seeley, D; Wolf, A.; Kafatos, F C

    1998-01-01

    Six gene markers have been used to map the progress of the innate immune response of the mosquito vector, Anopheles gambiae, upon infection by the malaria parasite, Plasmodium berghei. In addition to four previously reported genes, the set of markers included NOS (a nitric oxide synthase gene fragment) and ICHIT (a gene encoding two putative chitin-binding domains separated by a polythreonine-rich mucin region). In the midgut, a robust response occurs at 24 h post-infection, at a time when ma...

  4. 3-Oxoisoxazole-2(3H)-carboxamides and isoxazol-3-yl carbamates: Resistance-breaking acetylcholinesterase inhibitors targeting the malaria mosquito, Anopheles gambiae

    Science.gov (United States)

    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

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

    OpenAIRE

    Kayode David Ileke; Olaniyi Charles Ogungbite

    2015-01-01

    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 an...

  6. The presence of Plasmodium falciparum gametocytes in human blood increases the gravidity of Anopheles gambiae mosquitoes

    NARCIS (Netherlands)

    Ferguson, H.M.; Gouagna, L.C.; Obare, P.; Read, A.F.; Babiker, H.; Githure, J.I.; Beier, J.C.

    2005-01-01

    We conducted a field study in an area of endemic malaria transmission in western Kenya to determine whether mosquitoes that feed on gametocyte-infected blood but do not become infected have reduced or enhanced fecundity in comparison to mosquitoes fed on uninfected blood. Fifteen paired membrane-fee

  7. Identification and characterization of the fibrinogen-like domain of fibrinogen-related proteins in the mosquito, Anopheles gambiae, and the fruitfly, Drosophila melanogaster, genomes

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    Zhao Qin

    2005-09-01

    Full Text Available Abstract Background The fibrinogen-like (FBG domain, which consists of approximately 200 amino acid residues, has high sequence similarity to the C-terminal halves of fibrinogen β and γ chains. Fibrinogen-related proteins (FREPs, which contain FBG domains in their C-terminal region, are found universally in vertebrates and invertebrates. In invertebrates, FREPs are involved in immune responses and other aspects of physiology. To understand the complexity of this family in insects, we analyzed FREPs in the mosquito genome and made comparisons to FREPs in the fruitfly genome. Results By using the genome data of the mosquito, Anopheles gambiae, 53 FREPs were identified, whereas only 20 members were found in the Drosophila melanogaster genome. Using sequence profile analysis, we found that FBG domains have high sequence similarity and are highly conserved throughout the FBG domain region. By secondary structure analysis and comparison, the FBG domains of FREPs are predicted to function in recognition of carbohydrates and their derivatives on the surface of microorganisms in innate immunity. Conclusion Detailed sequence and structural analysis discloses that the FREP family contains FBG domains that have high sequence similarity in the A. gambiae genome. Expansion of the FREP family in mosquitoes during evolutionary history is mainly accounted for by a major expansion of the FBG domain architecture. The characterization of the FBG domains in the FREP family is likely to aid in the experimental analysis of the ability of mosquitoes to recognize parasites in innate immunity and physiologies associated with blood feeding.

  8. 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.

  9. Development of the bi-partite Gal4-UAS system in the African malaria mosquito, Anopheles gambiae.

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    Amy Lynd

    Full Text Available Functional genetic analysis in Anopheles gambiae would be greatly improved by the development of a binary expression system, which would allow the more rapid and flexible characterisation of genes influencing disease transmission, including those involved in insecticide resistance, parasite interaction, host and mate seeking behaviour. The Gal4-UAS system, widely used in Drosophila melanogaster functional genetics, has been significantly modified to achieve robust application in several different species. Towards this end, previous work generated a series of modified Gal4 constructs that were up to 20 fold more active than the native gene in An. gambiae cells. To examine the Gal4-UAS system in vivo, transgenic An. gambiae driver lines carrying a modified Gal4 gene under the control of the carboxypeptidase promoter, and responder lines carrying UAS regulated luciferase and eYFP reporter genes have been created. Crossing of the Gal4 and UAS lines resulted in progeny that expressed both reporters in the expected midgut specific pattern. Although there was minor variation in reporter gene activity between the different crosses examined, the tissue specific expression pattern was consistent regardless of the genomic location of the transgene cassettes. The results show that the modified Gal4-UAS system can be used to successfully activate expression of transgenes in a robust and tissue specific manner in Anopheles gambiae. The midgut driver and dual reporter responder constructs are the first to be developed and tested successfully in transgenic An. gambiae and provide the basis for further advancement of the system in this and other insect species.

  10. The immunogenic properties of protozoan glycosylphosphatidylinositols in the mosquito Anopheles gambiae.

    Science.gov (United States)

    Arrighi, Romanico B G; Debierre-Grockiego, Françoise; Schwarz, Ralph T; Faye, Ingrid

    2009-02-01

    In contrast to humans, mosquitoes do not have an adaptive immune response to deal with pathogens, and therefore must rely on their innate immune system to deal with invaders. This facilitates the recognition of different microbes on the basis of surface components or antigens. Such antigens have been identified in various types of microbe such as bacteria and fungi, yet none has been identified in the genus protozoa, which includes pathogens such as the malaria parasite, Plasmodium falciparum and Toxoplasma gondii. This study allowed us to test the antigenic properties of protozoan glycosylphosphatidylinositol (GPI) on the mosquito immune system. We found that both P. falciparum GPI and T. gondii GPI induce the strong expression of several antimicrobial peptides following ingestion, and that as a result of the immune response against the GPIs, the number of eggs produced by the mosquito is reduced dramatically. Such effects have been associated with malaria infected mosquitoes, but never associated with a Plasmodium specific antigen. This study demonstrates that protozoan GPIs can be considered as protozoan specific immune elicitors in mosquitoes, and that P. falciparum GPI plays a critical role in the malaria parasite manipulation of the mosquito vector to facilitate its transmission.

  11. Olfactory coding in antennal neurons of the malaria mosquito, Anopheles gambiae

    NARCIS (Netherlands)

    Qiu, Y.T.; Loon, van J.J.A.; Takken, W.; Meijerink, J.; Smid, H.M.

    2006-01-01

    Olfactory receptor neurons (ORNs) in the antenna of insects serve to encode odors in action potential activity conducted to the olfactory lobe of the deuterocerebrum. We performed an analysis of the electrophysiological responses of olfactory neurons in the antennae of the female malaria mosquito An

  12. 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

  13. CYP6 P450 enzymes and ACE-1 duplication produce extreme and multiple insecticide resistance in the malaria mosquito Anopheles gambiae.

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    Constant V Edi

    2014-03-01

    Full Text Available Malaria control relies heavily on pyrethroid insecticides, to which susceptibility is declining in Anopheles mosquitoes. To combat pyrethroid resistance, application of alternative insecticides is advocated for indoor residual spraying (IRS, and carbamates are increasingly important. Emergence of a very strong carbamate resistance phenotype in Anopheles gambiae from Tiassalé, Côte d'Ivoire, West Africa, is therefore a potentially major operational challenge, particularly because these malaria vectors now exhibit resistance to multiple insecticide classes. We investigated the genetic basis of resistance to the most commonly-applied carbamate, bendiocarb, in An. gambiae from Tiassalé. Geographically-replicated whole genome microarray experiments identified elevated P450 enzyme expression as associated with bendiocarb resistance, most notably genes from the CYP6 subfamily. P450s were further implicated in resistance phenotypes by induction of significantly elevated mortality to bendiocarb by the synergist piperonyl butoxide (PBO, which also enhanced the action of pyrethroids and an organophosphate. CYP6P3 and especially CYP6M2 produced bendiocarb resistance via transgenic expression in Drosophila in addition to pyrethroid resistance for both genes, and DDT resistance for CYP6M2 expression. CYP6M2 can thus cause resistance to three distinct classes of insecticide although the biochemical mechanism for carbamates is unclear because, in contrast to CYP6P3, recombinant CYP6M2 did not metabolise bendiocarb in vitro. Strongly bendiocarb resistant mosquitoes also displayed elevated expression of the acetylcholinesterase ACE-1 gene, arising at least in part from gene duplication, which confers a survival advantage to carriers of additional copies of resistant ACE-1 G119S alleles. Our results are alarming for vector-based malaria control. Extreme carbamate resistance in Tiassalé An. gambiae results from coupling of over-expressed target site allelic

  14. CYP6 P450 enzymes and ACE-1 duplication produce extreme and multiple insecticide resistance in the malaria mosquito Anopheles gambiae.

    Science.gov (United States)

    Edi, Constant V; Djogbénou, Luc; Jenkins, Adam M; Regna, Kimberly; Muskavitch, Marc A T; Poupardin, Rodolphe; Jones, Christopher M; Essandoh, John; Kétoh, Guillaume K; Paine, Mark J I; Koudou, Benjamin G; Donnelly, Martin J; Ranson, Hilary; Weetman, David

    2014-03-01

    Malaria control relies heavily on pyrethroid insecticides, to which susceptibility is declining in Anopheles mosquitoes. To combat pyrethroid resistance, application of alternative insecticides is advocated for indoor residual spraying (IRS), and carbamates are increasingly important. Emergence of a very strong carbamate resistance phenotype in Anopheles gambiae from Tiassalé, Côte d'Ivoire, West Africa, is therefore a potentially major operational challenge, particularly because these malaria vectors now exhibit resistance to multiple insecticide classes. We investigated the genetic basis of resistance to the most commonly-applied carbamate, bendiocarb, in An. gambiae from Tiassalé. Geographically-replicated whole genome microarray experiments identified elevated P450 enzyme expression as associated with bendiocarb resistance, most notably genes from the CYP6 subfamily. P450s were further implicated in resistance phenotypes by induction of significantly elevated mortality to bendiocarb by the synergist piperonyl butoxide (PBO), which also enhanced the action of pyrethroids and an organophosphate. CYP6P3 and especially CYP6M2 produced bendiocarb resistance via transgenic expression in Drosophila in addition to pyrethroid resistance for both genes, and DDT resistance for CYP6M2 expression. CYP6M2 can thus cause resistance to three distinct classes of insecticide although the biochemical mechanism for carbamates is unclear because, in contrast to CYP6P3, recombinant CYP6M2 did not metabolise bendiocarb in vitro. Strongly bendiocarb resistant mosquitoes also displayed elevated expression of the acetylcholinesterase ACE-1 gene, arising at least in part from gene duplication, which confers a survival advantage to carriers of additional copies of resistant ACE-1 G119S alleles. Our results are alarming for vector-based malaria control. Extreme carbamate resistance in Tiassalé An. gambiae results from coupling of over-expressed target site allelic variants with

  15. Fitness consequences of Anopheles gambiae population hybridization

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    Beier John C

    2005-09-01

    Full Text Available Abstract Background The use of transgenic mosquitoes with parasite inhibiting genes has been proposed as an integral strategy to control malaria transmission. However, release of exotic transgenic mosquitoes will bring in novel alleles along with parasite-inhibiting genes that may have unknown effects on native populations. Thus it is necessary to study the effects and dynamics of fitness traits in native mosquito populations in response to the introduction of novel genes. This study was designed to evaluate the dynamics of fitness traits in a simulation of introduction of novel alleles under laboratory conditions using two strains of Anopheles gambiae: Mbita strain from western Kenya and Ifakara strain from Tanzania. Methods The dynamics of fitness traits were evaluated under laboratory conditions using the two An. gambiae strains. These two geographically different strains were cross-bred and monitored for 20 generations to score fecundity, body size, blood-meal size, larval survival, and adult longevity, all of which are important determinants of the vector's potential in malaria transmission. Traits were analysed using pair-wise analysis of variance (ANOVA for fecundity, body size, and blood-meal size while survival analysis was performed for larval survival and adult longevity. Results Fecundity and body size were significantly higher in the progeny up to the 20th generation compared to founder strains. Adult longevity had a significantly higher mean up to the 10th generation and average blood-meal size was significantly larger up to the 5th generation, indicating that hybrids fitness is enhanced over that of the founder strains. Conclusion Hybridization of the two mosquito populations used in this study led to increased performance in the fitness traits studied. Given that the studied traits are important determinants of the vector's potential to transmit malaria, these results suggest the need to release genetically modified mosquitoes

  16. Infection of the malaria mosquito, Anopheles gambiae, with two species of entomopathogenic fungi: effects of concentration, co-formulation, exposure time and persistence

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    Lwetoijera Dickson W

    2009-12-01

    Full Text Available Abstract Background Entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana isolates have been shown to infect and reduce the survival of mosquito vectors. Methods Here four different bioassays were conducted to study the effect of conidia concentration, co-formulation, exposure time and persistence of the isolates M. anisopliae ICIPE-30 and B. bassiana I93-925 on infection and survival rates of female Anopheles gambiae sensu stricto. Test concentrations and exposure times ranged between 1 × 107 - 4 × 1010 conidia m-2 and 15 min - 6 h. In co-formulations, 2 × 1010 conidia m-2 of both fungus isolates were mixed at ratios of 4:1, 2:1, 1:1,1:0, 0:1, 1:2 and 1:4. To determine persistence, mosquitoes were exposed to surfaces treated 1, 14 or 28 d previously, with conidia concentrations of 2 × 109, 2 × 1010 or 4 × 1010. Results Mosquito survival varied with conidia concentration; 2 × 1010 conidia m-2 was the concentration above which no further reductions in survival were detectable for both isolates of fungus. The survival of mosquitoes exposed to single and co-formulated treatments was similar and no synergistic or additive effects were observed. Mosquitoes were infected within 30 min and longer exposure times did not result in a more rapid killing effect. Fifteen min exposure still achieved considerable mortality rates (100% mortality by 14 d of mosquitoes, but at lower speed than with 30 min exposure (100% mortality by 9 d. Conidia remained infective up to 28 d post-application but higher concentrations did not increase persistence. Conclusion Both fungus isolates are effective and persistent at low concentrations and short exposure times.

  17. Malaria infection of the mosquito Anopheles gambiae activates immune-responsive genes during critical transition stages of the parasite life cycle.

    Science.gov (United States)

    Dimopoulos, G; Seeley, D; Wolf, A; Kafatos, F C

    1998-01-01

    Six gene markers have been used to map the progress of the innate immune response of the mosquito vector, Anopheles gambiae, upon infection by the malaria parasite, Plasmodium berghei. In addition to four previously reported genes, the set of markers included NOS (a nitric oxide synthase gene fragment) and ICHIT (a gene encoding two putative chitin-binding domains separated by a polythreonine-rich mucin region). In the midgut, a robust response occurs at 24 h post-infection, at a time when malaria ookinetes traverse the midgut epithelium, but subsides at later phases of malaria development. In contrast, the salivary glands show no significant response at 24 h, but are activated in a prolonged late phase when sporozoites are released from the midgut into the haemolymph and invade the glands, between 10 and 25 days after blood feeding. Furthermore, the abdomen of the mosquito minus the midgut shows significant activation of immune markers, with complex kinetics that are distinct from those of both midgut and salivary glands. The parasite evidently elicits immune responses in multiple tissues of the mosquito, two of which are epithelia that the parasite must traverse to complete its development. The mechanisms of these responses and their significance for malaria transmission are discussed. PMID:9799221

  18. Vector competence of Anopheles and Culex mosquitoes for Zika virus

    Science.gov (United States)

    Dodson, Brittany L.

    2017-01-01

    Zika virus is a newly emergent mosquito-borne flavivirus that has caused recent large outbreaks in the new world, leading to dramatic increases in serious disease pathology including Guillain-Barre syndrome, newborn microcephaly, and infant brain damage. Although Aedes mosquitoes are thought to be the primary mosquito species driving infection, the virus has been isolated from dozens of mosquito species, including Culex and Anopheles species, and we lack a thorough understanding of which mosquito species to target for vector control. We exposed Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes to blood meals supplemented with two Zika virus strains. Mosquito bodies, legs, and saliva were collected five, seven, and 14 days post blood meal and tested for infectious virus by plaque assay. Regardless of titer, virus strain, or timepoint, Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes were refractory to Zika virus infection. We conclude that Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes likely do not contribute significantly to Zika virus transmission to humans. However, future studies should continue to explore the potential for other novel potential vectors to transmit the virus. PMID:28316896

  19. Vector competence of Anopheles and Culex mosquitoes for Zika virus

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    Brittany L. Dodson

    2017-03-01

    Full Text Available Zika virus is a newly emergent mosquito-borne flavivirus that has caused recent large outbreaks in the new world, leading to dramatic increases in serious disease pathology including Guillain-Barre syndrome, newborn microcephaly, and infant brain damage. Although Aedes mosquitoes are thought to be the primary mosquito species driving infection, the virus has been isolated from dozens of mosquito species, including Culex and Anopheles species, and we lack a thorough understanding of which mosquito species to target for vector control. We exposed Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes to blood meals supplemented with two Zika virus strains. Mosquito bodies, legs, and saliva were collected five, seven, and 14 days post blood meal and tested for infectious virus by plaque assay. Regardless of titer, virus strain, or timepoint, Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes were refractory to Zika virus infection. We conclude that Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes likely do not contribute significantly to Zika virus transmission to humans. However, future studies should continue to explore the potential for other novel potential vectors to transmit the virus.

  20. Vector competence of Anopheles and Culex mosquitoes for Zika virus.

    Science.gov (United States)

    Dodson, Brittany L; Rasgon, Jason L

    2017-01-01

    Zika virus is a newly emergent mosquito-borne flavivirus that has caused recent large outbreaks in the new world, leading to dramatic increases in serious disease pathology including Guillain-Barre syndrome, newborn microcephaly, and infant brain damage. Although Aedes mosquitoes are thought to be the primary mosquito species driving infection, the virus has been isolated from dozens of mosquito species, including Culex and Anopheles species, and we lack a thorough understanding of which mosquito species to target for vector control. We exposed Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes to blood meals supplemented with two Zika virus strains. Mosquito bodies, legs, and saliva were collected five, seven, and 14 days post blood meal and tested for infectious virus by plaque assay. Regardless of titer, virus strain, or timepoint, Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes were refractory to Zika virus infection. We conclude that Anopheles gambiae, Anopheles stephensi, and Culex quinquefasciatus mosquitoes likely do not contribute significantly to Zika virus transmission to humans. However, future studies should continue to explore the potential for other novel potential vectors to transmit the virus.

  1. 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

  2. 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

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

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    Smallegange, Renate C; Qiu, Yu Tong; Bukovinszkiné-Kiss, Gabriella; Van Loon, Joop J A; Takken, Willem

    2009-08-01

    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 were tested in a tripartite blend with ammonia + lactic acid. Four different airflow stream rates (0.5, 5, 50, and 100 ml/min) carrying the compounds were tested for their effect on trap entry response in the olfactometer. In the olfactometer, propanoic acid, butanoic acid, 3-methylbutanoic acid, pentanoic acid, heptanoic acid, octanoic acid, and tetradecanoic acid increased attraction relative to the basic blend. While several carboxylic acids were attractive only at one or two flow rates, tetradecanoic acid was attractive at all flow rates tested. Heptanoic acid was attractive at the lowest flow rate (0.5 ml/min), but repellent at 5 and 50 ml/min. Mixing the air stream laden with these 7 carboxylic acids together with the headspace of the basic blend increased attraction in two quantitative compositions. Subtraction of single acids from the most attractive blend revealed that 3-methylbutanoic acid had a negative effect on trap entry response. In the absence of tetradecanoic acid, the blend was repellent. In assays with MM-X traps, both a blend of 7 carboxylic acids + ammonia + lactic acid (all applied from low density polyethylene-sachets) and a simple blend of ammonia + lactic acid + tetradecanoic acid were attractive. The results show that carboxylic acids play an essential role in the host-seeking behavior of An. gambiae, and that the contribution to blend attractiveness depends on the specific compound studied.

  4. 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

  5. High Genetic Differentiation between the M and S Molecular Forms of Anopheles gambiae in Africa

    OpenAIRE

    Caroline Esnault; Matthieu Boulesteix; Jean Bernard Duchemin; Koffi, Alphonsine A; Fabrice Chandre; Roch Dabiré; Vincent Robert; Frédéric Simard; Frédéric Tripet; Donnelly, Martin J.; Didier Fontenille; Christian Biémont

    2008-01-01

    BACKGROUND: Anopheles gambiae, a major vector of malaria, is widely distributed throughout sub-Saharan Africa. In an attempt to eliminate infective mosquitoes, researchers are trying to develop transgenic strains that are refractory to the Plasmodium parasite. Before any release of transgenic mosquitoes can be envisaged, we need an accurate picture of the differentiation between the two molecular forms of An. gambiae, termed M and S, which are of uncertain taxonomic status. METHODOLOGY/PRINCI...

  6. Genomic islands of speciation in Anopheles gambiae.

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    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

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

    Science.gov (United States)

    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...

  8. Composition of Anopheles coluzzii and Anopheles gambiae microbiota from larval to adult stages.

    Science.gov (United States)

    Gimonneau, Geoffrey; Tchioffo, Majoline T; Abate, Luc; Boissière, Anne; Awono-Ambéné, Parfait H; Nsango, Sandrine E; Christen, Richard; Morlais, Isabelle

    2014-12-01

    During their immature life stages, malaria mosquitoes are exposed to a wide array of microbes and contaminants from the aquatic habitats. Although prior studies have suggested that environmental exposure shapes the microbial community structure in the adult mosquito, most reports have focused on laboratory-based experiments and on a single mosquito epithelium, the gut. In this study, we investigated the influence of the breeding site on the development of the Anopheles coluzzii and Anopheles gambiae microbiota in natural conditions. We characterized bacterial communities from aquatic habitats, at surface microlayer and subsurface water levels, to freshly emerge adult mosquitoes using multiplexed 16S rRNA gene pyrosequencing and we separately analyzed the microbiota associated with the different epithelia of adult individual, midguts, ovaries and salivary glands. We found that the distribution of bacterial communities in the aquatic habitats differed according to the depth of water collections. Inter-individual variation of bacterial composition was large in larvae guts but adult mosquitoes from a same breeding site shared quite similar microbiota. Although some differences in bacterial abundances were highlighted between the different epithelia of freshly emerged An. coluzzii and An. gambiae, an intriguing feature from our study is the particular similarity of the overall bacterial communities. Our results call for further investigations on the bacterial population dynamics in the different tissues to determine the distinctive characteristics of each microbiota during the mosquito lifespan and to identify specific interactions between certain key phyla or species and the insect life history traits.

  9. In vitro and in vivo host range of Anopheles gambiae densovirus (AgDNV).

    Science.gov (United States)

    Suzuki, Yasutsugu; Barik, Tapan K; Johnson, Rebecca M; Rasgon, Jason L

    2015-07-29

    AgDNV is a powerful gene transduction tool and potential biological control agent for Anopheles mosquitoes. Using a GFP reporter virus system, we investigated AgDNV host range specificity in four arthropod cell lines (derived from An. gambiae, Aedes albopictus and Drosophila melanogaster) and six mosquito species from 3 genera (An. gambiae, An. arabiensis, An. stephensi, Ae. albopictus, Ae. aegypti and Culex tarsalis). In vitro, efficient viral invasion, replication and GFP expression was only observed in MOS55 An. gambiae cells. In vivo, high levels of GFP were observed in An. gambiae mosquitoes. Intermediate levels of GFP were observed in the closely related species An. arabiensis. Low levels of GFP were observed in An. stephensi, Ae. albopictus, Ae. aegypti and Cx. tarsalis. These results suggest that AgDNV is a specific gene transduction tool for members of the An. gambiae species complex, and could be potentially developed into a biocontrol agent with minimal off-target effects.

  10. Chloroquine mediated modulation of Anopheles gambiae gene expression.

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    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.

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

    NARCIS (Netherlands)

    Farenhorst, M.; Knols, B.G.J.; Thomas, M.B.; Howard, A.F.V.; Takken, W.; Rowland, M.; N'Guessan, R.

    2010-01-01

    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 invest

  12. Susceptibility of Anopheles gambiae and Anopheles stephensi to tropical isolates of Plasmodium falciparum

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    Ranford-Cartwright Lisa C

    2007-10-01

    Full Text Available Abstract Background The susceptibility of anopheline mosquito species to Plasmodium infection is known to be variable with some mosquitoes more permissive to infection than others. Little work, however, has been carried out investigating the susceptibility of major malaria vectors to geographically diverse tropical isolates of Plasmodium falciparum aside from examining the possibility of infection extending its range from tropical regions into more temperate zones. Methods This study investigates the susceptibility of two major tropical mosquito hosts (Anopheles gambiae and Anopheles stephensi to P. falciparum isolates of different tropical geographical origins. Cultured parasite isolates were fed via membrane feeders simultaneously to both mosquito species and the resulting mosquito infections were compared. Results Infection prevalence was variable with African parasites equally successful in both mosquito species, Thai parasites significantly more successful in An. stephensi, and PNG parasites largely unsuccessful in both species. Conclusion Infection success of P. falciparum was variable according to geographical origin of both the parasite and the mosquito. Data presented raise the possibility that local adaptation of tropical parasites and mosquitoes has a role to play in limiting gene flow between allopatric parasite populations.

  13. 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

  14. Anopheles gambiae immune responses to human and rodent Plasmodium parasite species.

    OpenAIRE

    Yuemei Dong; Ruth Aguilar; Zhiyong Xi; Emma Warr; Emmanuel Mongin; George Dimopoulos

    2006-01-01

    Transmission of malaria is dependent on the successful completion of the Plasmodium lifecycle in the Anopheles vector. Major obstacles are encountered in the midgut tissue, where most parasites are killed by the mosquito's immune system. In the present study, DNA microarray analyses have been used to compare Anopheles gambiae responses to invasion of the midgut epithelium by the ookinete stage of the human pathogen Plasmodium falciparum and the rodent experimental model pathogen P. berghei. I...

  15. Feeding and survival of the malaria vector Anopheles gambiae on plants growing in Kenia

    NARCIS (Netherlands)

    Impoinvil, D.E.; Kongere, J.O.; Foster, W.A.; Njiru, B.N.; Killeen, G.F.; Githure, J.I.; Beier, J.C.; Hassanali, A.; Knols, B.G.J.

    2004-01-01

    The propensity of the malaria vector mosquito Anopheles gambiae Giles (Diptera: Culicidae) to ingest sugars from various plants, and subsequent survival rates, were assessed with laboratory-reared males and females offered eight species of plants commonly cultivated and/or growing wild in western Ke

  16. Factors affecting the vectorial competence of Anopheles gambiae: a question of scale

    NARCIS (Netherlands)

    Takken, W.; Lindsay, S.W.

    2003-01-01

    Malaria transmission in Africa is without doubt governed by the existence of a group of highly efficient vectors, of which Anopheles gambiae Giles sensu stricto is predominant. The endophilic and anthropophagic behaviours of this mosquito create an intimate association between the human reservoir an

  17. 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

  18. 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.

  19. The dance of male Anopheles gambiae in wild mating swarms.

    Science.gov (United States)

    Butail, Sachit; Manoukis, Nicholas C; Diallo, Moussa; Ribeiro, José M C; Paley, Derek A

    2013-05-01

    An important element of mating in the malaria vector Anopheles gambiae Giles in nature is the crepuscular mating aggregation (swarm) composed almost entirely of males, where most coupling and insemination is generally believed to occur. In this study, we mathematically characterize the oscillatory movement of male An. gambiae in terms of an established individual-based mechanistic model that parameterizes the attraction of a mosquito toward the center of the swarm using the natural frequency of oscillation and the resistance to its motion, characterized by the damping ratio. Using three-dimensional trajectory data of ten wild mosquito swarms filmed in Mali, Africa, we show two new results for low and moderate wind conditions, and indicate how these results may vary in high wind. First, we show that in low and moderate wind the vertical component of the mosquito motion has a lower frequency of oscillation and higher damping ratio than horizontal motion. In high wind, the vertical and horizontal motions are similar to one another and the natural frequencies are higher than in low and moderate wind. Second, we show that the predicted average disagreement in the direction of motion of swarming mosquitoes moving randomly is greater than the average disagreement we observed between each mosquito and its three closest neighbors, with the smallest level of disagreement occurring for the nearest neighbor in seven out of 10 swarms. The alignment of the direction of motion between nearest neighbors is the highest in high wind. This result provides evidence for flight-path coordination between swarming male mosquitoes.

  20. 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

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

    Directory of Open Access Journals (Sweden)

    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.

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

    Directory of Open Access Journals (Sweden)

    Nazaire Aïzoun

    2014-04-01

    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 physiological status. Therefore, it is useful to respect the World Health Organization criteria in the assessment of insecticide susceptibility tests in malaria vectors. Otherwise, susceptibility testing is conducted using unfed female mosquitoes aged 3-5 days old. Tests should also be carried out at (25±2 °C and (80±10% relative humidity.

  3. Analysis of apyrase 5' upstream region validates improved Anopheles gambiae transformation technique

    Directory of Open Access Journals (Sweden)

    Lanfrancotti Alessandra

    2009-02-01

    Full Text Available Abstract Background Genetic transformation of the malaria mosquito Anopheles gambiae has been successfully achieved in recent years, and represents a potentially powerful tool for researchers. Tissue-, stage- and sex-specific promoters are essential requirements to support the development of new applications for the transformation technique and potential malaria control strategies. During the Plasmodium lifecycle in the invertebrate host, four major mosquito cell types are involved in interactions with the parasite: hemocytes and fat body cells, which provide humoral and cellular components of the innate immune response, midgut and salivary glands representing the epithelial barriers traversed by the parasite during its lifecycle in the mosquito. Findings We have analyzed the upstream regulatory sequence of the An. gambiae salivary gland-specific apyrase (AgApy gene in transgenic An. gambiae using a piggyBac transposable element vector marked by a 3xP3 promoter:DsRed gene fusion. Efficient germ-line transformation in An. gambiae mosquitoes was obtained and several integration events in at least three different G0 families were detected. LacZ reporter gene expression was analyzed in three transgenic lines/groups, and in only one group was tissue-specific expression restricted to salivary glands. Conclusion Our data describe an efficient genetic transformation of An. gambiae embryos. However, expression from the selected region of the AgApy promoter is weak and position effects may mask tissue- and stage- specific activity in transgenic mosquitoes.

  4. Evidence for Population-Specific Positive Selection on Immune Genes of Anopheles gambiae

    OpenAIRE

    Crawford, Jacob E; Bischoff, Emmanuel; Garnier, Thierry; Gneme, Awa; Eiglmeier, Karin; Holm, Inge; Michelle M Riehle; Guelbeogo, Wamdaogo M.; Sagnon, N’Fale; Brian P Lazzaro; Vernick, Kenneth D

    2012-01-01

    Host-pathogen interactions can be powerful drivers of adaptive evolution, shaping the patterns of molecular variation at the genes involved. In this study, we sequenced alleles from 28 immune-related loci in wild samples of multiple genetic subpopulations of the African malaria mosquito Anopheles gambiae, obtaining unprecedented sample sizes and providing the first opportunity to contrast patterns of molecular evolution at immune-related loci in the recently discovered GOUNDRY population to t...

  5. Larvicidal efficacy of monoterpenes against the larvae of Anopheles gambiae

    Institute of Scientific and Technical Information of China (English)

    Eliningaya J.Kweka; Tamires Cardoso Lima; Chrian M.Marciale; Dami?o Pergentino de Sousa

    2016-01-01

    Objective:To evaluate the larvicidal efficacy of eight volatile components of essential oils against 3rd instar larvae of Anopheles gambiae s.s.Methods:Larvicidal effects of each compound were evaluated in both laboratory and semi-field trials.Stock solution was prepared and serial dilutions were made in six concentrations for each compound.A total of 20 larvae were exposed to larvicides for each replicate and monitored at intervals of 12,24,48 and 72 h.Larvae monitoring was done on basis of dead and live larvae in all intervals.Results:All assayed compounds were larvicides and presented varying degrees of larval toxicity,with LC50 values ranging from 1.28 to 1 938.92 mg/L depending on the treatment time(12,24,48 or 72 h).(-)-Perillyl alcohol presented the strongest larvicidal activity towards Anopheles gambiae larvae,with LC50 values of 73.60,18.36,1.72 and1.28 mg/L after 12,24,48 and 72 h of exposure,respectively.The next strongest were(-)-isopulegol(LC50= 135.10,49.39,34.39 and 20.22 mg/L) and(-)-carvone epoxide(LC50= 168.86,124.74,80.84 and 23.46 mg/L).After 12,24 and 48 h of treatment,hydroxydihydrocarvone was the least toxic compound,with LC50 values of 1 938.92,1 172.18 and 401.03 mg/L,respectively.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.

  6. 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

    Directory of Open Access Journals (Sweden)

    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

  7. 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

  8. Odour sensitivity of antennal olfactory cells underlying grooved pegs of Anopheles gambiae s.s. and An. quadriannulatus

    NARCIS (Netherlands)

    van den Broek, IVF; den Otter, CJ

    2000-01-01

    In female mosquitoes of the anthropophilic species Anopheles gambiae Giles s.s. and the zoophilic An. quadriannulatus Theobald single sensillum recordings from grooved pegs were made. In both species, the majority of these sensilla responded to ammonium hydroxide, butylamine and propanoic acid, wher

  9. Use of Near-Infrared Spectroscopy to Age-Grade and Identify Siblings of Anopheles Gambiae Complex

    Science.gov (United States)

    We used near-infrared spectroscopy (NIRS) to rapidly and non-destructively determine species and age of Anopheles gambiae ss (G3, Mali-NIH, Kisumu, ZANU, and Ifakara strains) and An. arabiensis (Dongola, KGB, and Ifakara strains). We developed NIR calibrations using mosquitoes reared and scanned at ...

  10. Reticulate Speciation and Barriers to Introgression in the Anopheles gambiae Species Complex.

    Science.gov (United States)

    Crawford, Jacob E; Riehle, Michelle M; Guelbeogo, Wamdaogo M; Gneme, Awa; Sagnon, N'Fale; Vernick, Kenneth D; Nielsen, Rasmus; Lazzaro, Brian P

    2015-11-28

    Speciation as a process remains a central focus of evolutionary biology, but our understanding of the genomic architecture and prevalence of speciation in the face of gene flow remains incomplete. The Anopheles gambiae species complex of malaria mosquitoes is a radiation of ecologically diverse taxa. This complex is well-suited for testing for evidence of a speciation continuum and genomic barriers to introgression because its members exhibit partially overlapping geographic distributions as well as varying levels of divergence and reproductive isolation. We sequenced 20 genomes from wild A. gambiae s.s., Anopheles coluzzii, Anopheles arabiensis, and compared these with 12 genomes from the "GOUNDRY" subgroup of A. gambiae s.l. Amidst a backdrop of strong reproductive isolation, we find strong evidence for a speciation continuum with introgression of autosomal chromosomal regions among species and subgroups. The X chromosome, however, is strongly differentiated among all taxa, pointing to a disproportionately large effect of X chromosome genes in driving speciation among anophelines. Strikingly, we find that autosomal introgression has occurred from contemporary hybridization between A. gambiae and A. arabiensis despite strong divergence (∼5× higher than autosomal divergence) and isolation on the X chromosome. In addition to the X, we find strong evidence that lowly recombining autosomal regions, especially pericentromeric regions, serve as barriers to introgression secondarily to the X. We show that speciation with gene flow results in genomic mosaicism of divergence and introgression. Such a reticulate gene pool connecting vector taxa across the speciation continuum has important implications for malaria control efforts.

  11. Dissecting the organ specificity of insecticide resistance candidate genes in Anopheles gambiae: known and novel candidate genes

    OpenAIRE

    2014-01-01

    Background The elevated expression of enzymes with insecticide metabolism activity can lead to high levels of insecticide resistance in the malaria vector, Anopheles gambiae. In this study, adult female mosquitoes from an insecticide susceptible and resistant strain were dissected into four different body parts. RNA from each of these samples was used in microarray analysis to determine the enrichment patterns of the key detoxification gene families within the mosquito and to identify additio...

  12. Genetic population structure of Anopheles gambiae in Equatorial Guinea

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    Caccone Adalgisa

    2007-10-01

    Full Text Available Abstract Background Patterns of genetic structure among mosquito vector populations in islands have received particular attention as these are considered potentially suitable sites for experimental trials on transgenic-based malaria control strategies. In this study, levels of genetic differentiation have been estimated between populations of Anopheles gambiae s.s. from the islands of Bioko and Annobón, and from continental Equatorial Guinea (EG and Gabon. Methods Genotyping of 11 microsatellite loci located in chromosome 3 was performed in three island samples (two in Bioko and one in Annobón and three mainland samples (two in EG and one in Gabon. Four samples belonged to the M molecular form and two to the S-form. Microsatellite data was used to estimate genetic diversity parameters, perform demographic equilibrium tests and analyse population differentiation. Results High levels of genetic differentiation were found between the more geographically remote island of Annobón and the continent, contrasting with the shallow differentiation between Bioko island, closest to mainland, and continental localities. In Bioko, differentiation between M and S forms was higher than that observed between island and mainland samples of the same molecular form. Conclusion The observed patterns of population structure seem to be governed by the presence of both physical (the ocean and biological (the M-S form discontinuity barriers to gene flow. The significant degree of genetic isolation between M and S forms detected by microsatellite loci located outside the "genomic islands" of speciation identified in A. gambiae s.s. further supports the hypothesis of on-going incipient speciation within this species. The implications of these findings regarding vector control strategies are discussed.

  13. 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......, in an area of north-eastern Tanzania which has been exposed to ivermectin mass drug administration, this study assessed and compared the effect of human ivermectin treatment on blood feeding Anopheles gambiae and Cx. quinquefasciatus. METHODS: Consenting adult volunteers were randomized into two groups...... 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...

  14. Pyrethroid and DDT Resistance and Organophosphate Susceptibility among Anopheles spp. Mosquitoes, Western Kenya.

    Science.gov (United States)

    Wanjala, Christine L; Mbugi, Jernard P; Ototo, Edna; Gesuge, Maxwell; Afrane, Yaw A; Atieli, Harrysone E; Zhou, Guofa; Githeko, Andrew K; Yan, Guiyun

    2015-12-01

    We conducted standard insecticide susceptibility testing across western Kenya and found that the Anopheles gambiae mosquito has acquired high resistance to pyrethroids and DDT, patchy resistance to carbamates, but no resistance to organophosphates. Use of non-pyrethroid-based vector control tools may be preferable for malaria prevention in this region.

  15. 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.

  16. The practical importance of permanent and semipermanent habitats for controlling aquatic stages of Anopheles gambiae sensu lato mosquitoes: operational observations from a rural town in western Kenya

    NARCIS (Netherlands)

    Fillinger, U.; Sonye, G.; Killeen, G.F.; Knols, B.G.J.; Becker, N.

    2004-01-01

    Control of aquatic-stage Anopheles is one of the oldest and most historically successful interventions to prevent malaria, but it has seen little application in Africa. Consequently, the ecology of immature afrotropical Anopheles has received insufficient attention. We therefore examined the populat

  17. 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.

  18. 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.

  19. 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

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

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    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

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

    Directory of Open Access Journals (Sweden)

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

    Directory of Open Access Journals (Sweden)

    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

  3. A 3D Analysis of Flight Behavior of Anopheles gambiae sensu stricto Malaria Mosquitoes in Response to Human Odor and Heat

    NARCIS (Netherlands)

    Spitzen, J.; Spoor, C.W.; Grieco, F.; Braak, ter C.J.F.; Beeuwkes, J.; Brugge, van S.P.; Kranenbarg, S.; Noldus, L.; Leeuwen, van J.L.; Takken, W.

    2013-01-01

    Female mosquitoes use odor and heat as cues to navigate to a suitable landing site on their blood host. The way these cues affect flight behavior and modulate anemotactic responses, however, is poorly understood. We studied in-flight behavioral responses of females of the nocturnal malaria mosquito

  4. Effects of a botanical larvicide derived from Azadirachta indica (the neem tree) on oviposition behaviour in Anopheles gambiae s.s. mosquitoes

    NARCIS (Netherlands)

    Howard, A.F.V.; Adongo, E.A.; Vulule, J.; Githure, J.

    2011-01-01

    More focus is given to mosquito larval control due to the necessity to use several control techniques together in integrated vector management programmes. Botanical products are thought to be able to provide effective, sustainable and cheap mosquito larval control tools. However, bio-larvicides like

  5. Optimizing odor-baited trap methods for collecting mosquitoes during the malaria season in The Gambia.

    Directory of Open Access Journals (Sweden)

    Musa Jawara

    Full Text Available BACKGROUND: Baited traps are potential tools for removal or surveillance of disease vectors. To optimize the use of counter-flow traps baited with human odor (nylon socks that had been worn for a single day to capture wild mosquitoes in the Gambia, investigations were conducted at a field experimental site. METHODOLOGY/PRINCIPAL FINDINGS: Experiments employing Latin square design were conducted with a set of six huts to investigate the effects of the following on overnight mosquito trap catches: (1 placement of traps indoors or immediately outdoors, CO(2 supply, and presence of a human subject in the hut; (2 trap height for collecting mosquitoes immediately outdoors; (3 height and distance from hut; (4 interaction between multiple traps around a single hut and entry of mosquitoes into huts. A total of 106,600 adult mosquitoes (9.1% Anopheles gambiae s.l., 4.0% other Anopheles species were collected over 42 nights. The high numbers of An. gambiae s.l. and other mosquitoes collected by odor-baited traps required CO(2 but were largely independent of the presence of a person sleeping in the hut or of trap placement indoors or outdoors. For outdoor collection that is considered less intrusive, traps opening 15 cm above the floor of the hut veranda were more highly effective than traps at other heights or further from the hut. There was no significant evidence of saturation or competition by the traps, with multiple traps around a hut each collecting almost as many mosquitoes as single traps and no effect on the numbers of mosquitoes entering the huts. CONCLUSIONS/SIGNIFICANCE: The outdoor trapping protocol is convenient to compare attractiveness of different odors or synthetic chemicals to malaria vectors and other wild mosquitoes. The finding that such traps are reliably attractive in the presence or absence of a human volunteer encourages their potential development as standardised surveillance tools.

  6. 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

  7. 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.

  8. 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

    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......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....... Methods: A total of 1800 laboratory reared mosquito larvae of each species were used in the bioassays. Twelve replicates were performed, each testing 6 concentrations of ivermectin (0.0, 0.001, 0.01, 0.1, 1.0 and 10.0 parts per million (ppm)) against third instar larvae of An. gambiae and Cx...

  9. 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.

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

    Science.gov (United States)

    Ramos, Susana; Custódio, Ana; Silveira, Henrique

    2014-08-01

    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.

  11. 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.

  12. The Anopheles gambiae glutathione transferase supergene family: annotation, phylogeny and expression profiles

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    Rossiter Louise C

    2003-08-01

    Full Text Available Abstract Background Twenty-eight genes putatively encoding cytosolic glutathione transferases have been identified in the Anopheles gambiae genome. We manually annotated these genes and then confirmed the annotation by sequencing of A. gambiae cDNAs. Phylogenetic analysis with the 37 putative GST genes from Drosophila and representative GSTs from other taxa was undertaken to develop a nomenclature for insect GSTs. The epsilon class of insect GSTs has previously been implicated in conferring insecticide resistance in several insect species. We compared the expression level of all members of this GST class in two strains of A. gambiae to determine whether epsilon GST expression is correlated with insecticide resistance status. Results Two A. gambiae GSTs are alternatively spliced resulting in a maximum number of 32 transcripts encoding cytosolic GSTs. We detected cDNAs for 31 of these in adult mosquitoes. There are at least six different classes of GSTs in insects but 20 of the A. gambiae GSTs belong to the two insect specific classes, delta and epsilon. Members of these two GST classes are clustered on chromosome arms 2L and 3R respectively. Two members of the GST supergene family are intronless. Amongst the remainder, there are 13 unique introns positions but within the epsilon and delta class, there is considerable conservation of intron positions. Five of the eight epsilon GSTs are overexpressed in a DDT resistant strain of A. gambiae. Conclusions The GST supergene family in A. gambiae is extensive and regulation of transcription of these genes is complex. Expression profiling of the epsilon class supports earlier predictions that this class is important in conferring insecticide resistance.

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

    Science.gov (United States)

    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

  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. Infection of the malaria mosquito, Anopheles gambiae, with two species of entomopathogenic fungi: effects of concentration, co-formulation, exposure time and persistence

    NARCIS (Netherlands)

    Mnyone, L.L.; Kirby, M.J.; Lwetoijera, D.W.; Mpingwa, M.W.; Knols, B.G.J.; Takken, W.; Russell, T.L.

    2009-01-01

    Background - Entomopathogenic fungi Metarhizium anisopliae and Beauveria bassiana isolates have been shown to infect and reduce the survival of mosquito vectors. Methods - Here four different bioassays were conducted to study the effect of conidia concentration, co-formulation, exposure time and per

  16. Anopheles gambiae immune responses to human and rodent Plasmodium parasite species.

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    Yuemei Dong

    2006-06-01

    Full Text Available Transmission of malaria is dependent on the successful completion of the Plasmodium lifecycle in the Anopheles vector. Major obstacles are encountered in the midgut tissue, where most parasites are killed by the mosquito's immune system. In the present study, DNA microarray analyses have been used to compare Anopheles gambiae responses to invasion of the midgut epithelium by the ookinete stage of the human pathogen Plasmodium falciparum and the rodent experimental model pathogen P. berghei. Invasion by P. berghei had a more profound impact on the mosquito transcriptome, including a variety of functional gene classes, while P. falciparum elicited a broader immune response at the gene transcript level. Ingestion of human malaria-infected blood lacking invasive ookinetes also induced a variety of immune genes, including several anti-Plasmodium factors. Twelve selected genes were assessed for effect on infection with both parasite species and bacteria using RNAi gene silencing assays, and seven of these genes were found to influence mosquito resistance to both parasite species. An MD2-like receptor, AgMDL1, and an immunolectin, FBN39, showed specificity in regulating only resistance to P. falciparum, while the antimicrobial peptide gambicin and a novel putative short secreted peptide, IRSP5, were more specific for defense against the rodent parasite P. berghei. While all the genes that affected Plasmodium development also influenced mosquito resistance to bacterial infection, four of the antimicrobial genes had no effect on Plasmodium development. Our study shows that the impact of P. falciparum and P. berghei infection on A. gambiae biology at the gene transcript level is quite diverse, and the defense against the two Plasmodium species is mediated by antimicrobial factors with both universal and Plasmodium-species specific activities. Furthermore, our data indicate that the mosquito is capable of sensing infected blood constituents in the absence

  17. Isolation and characterization of native Bacillus thuringiensis strains from Saudi Arabia with enhanced larvicidal toxicity against the mosquito vector Anopheles gambiae (s.l.).

    OpenAIRE

    El-Kersh, TA; Ahmed, AM; Al-Sheikh, YA; Tripet, F; Ibrahim, MS; Metwalli, AAM

    2016-01-01

    BACKGROUND: Worldwide, mosquito vectors are transmitting several etiological agents of important human diseases, including malaria, causing millions of deaths every year. In Saudi Arabia, as elsewhere, vector-control is based mostly on chemical insecticides which may be toxic and cause environmental deprivation. Here, to support the development of bio-pesticide alternatives, a study was conducted to identify native Bacillus thuringiensis (Bt) isolates with improved toxicity against the malari...

  18. Isolation and characterization of native Bacillus thuringiensis strains from Saudi Arabia with enhanced larvicidal toxicity against the mosquito vector Anopheles gambiae (s.l.)

    OpenAIRE

    El-kersh, Talaat A.; Ahmed, Ashraf M.; AL-SHEIKH, YAZEED A.; Tripet, Frédéric; Ibrahim, Mohamed S.; Metwalli, Ali A. M.

    2016-01-01

    Background Worldwide, mosquito vectors are transmitting several etiological agents of important human diseases, including malaria, causing millions of deaths every year. In Saudi Arabia, as elsewhere, vector-control is based mostly on chemical insecticides which may be toxic and cause environmental deprivation. Here, to support the development of bio-pesticide alternatives, a study was conducted to identify native Bacillus thuringiensis (Bt) isolates with improved toxicity against the malaria...

  19. High genetic differentiation between the M and S molecular forms of Anopheles gambiae in Africa.

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    Caroline Esnault

    Full Text Available BACKGROUND: Anopheles gambiae, a major vector of malaria, is widely distributed throughout sub-Saharan Africa. In an attempt to eliminate infective mosquitoes, researchers are trying to develop transgenic strains that are refractory to the Plasmodium parasite. Before any release of transgenic mosquitoes can be envisaged, we need an accurate picture of the differentiation between the two molecular forms of An. gambiae, termed M and S, which are of uncertain taxonomic status. METHODOLOGY/PRINCIPAL FINDINGS: Insertion patterns of three transposable elements (TEs were determined in populations from Benin, Burkina Faso, Cameroon, Ghana, Ivory Coast, Madagascar, Mali, Mozambique, Niger, and Tanzania, using Transposon Display, a TE-anchored strategy based on Amplified Fragment Length Polymorphism. The results reveal a clear differentiation between the M and S forms, whatever their geographical origin, suggesting an incipient speciation process. CONCLUSIONS/SIGNIFICANCE: Any attempt to control the transmission of malaria by An. gambiae using either conventional or novel technologies must take the M/S genetic differentiation into account. In addition, we localized three TE insertion sites that were present either in every individual or at a high frequency in the M molecular form. These sites were found to be located outside the chromosomal regions that are suspected of involvement in the speciation event between the two forms. This suggests that these chromosomal regions are either larger than previously thought, or there are additional differentiated genomic regions interspersed with undifferentiated regions.

  20. High Genetic Differentiation between the M and S Molecular Forms of Anopheles gambiae in Africa

    Science.gov (United States)

    Esnault, Caroline; Boulesteix, Matthieu; Duchemin, Jean Bernard; Koffi, Alphonsine A.; Chandre, Fabrice; Dabiré, Roch; Robert, Vincent; Simard, Frédéric; Tripet, Frédéric; Donnelly, Martin J.; Fontenille, Didier; Biémont, Christian

    2008-01-01

    Background Anopheles gambiae, a major vector of malaria, is widely distributed throughout sub-Saharan Africa. In an attempt to eliminate infective mosquitoes, researchers are trying to develop transgenic strains that are refractory to the Plasmodium parasite. Before any release of transgenic mosquitoes can be envisaged, we need an accurate picture of the differentiation between the two molecular forms of An. gambiae, termed M and S, which are of uncertain taxonomic status. Methodology/Principal Findings Insertion patterns of three transposable elements (TEs) were determined in populations from Benin, Burkina Faso, Cameroon, Ghana, Ivory Coast, Madagascar, Mali, Mozambique, Niger, and Tanzania, using Transposon Display, a TE-anchored strategy based on Amplified Fragment Length Polymorphism. The results reveal a clear differentiation between the M and S forms, whatever their geographical origin, suggesting an incipient speciation process. Conclusions/Significance Any attempt to control the transmission of malaria by An. gambiae using either conventional or novel technologies must take the M/S genetic differentiation into account. In addition, we localized three TE insertion sites that were present either in every individual or at a high frequency in the M molecular form. These sites were found to be located outside the chromosomal regions that are suspected of involvement in the speciation event between the two forms. This suggests that these chromosomal regions are either larger than previously thought, or there are additional differentiated genomic regions interspersed with undifferentiated regions. PMID:18414665

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

    Science.gov (United States)

    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

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

    Directory of Open Access Journals (Sweden)

    Lyimo Issa N

    2009-12-01

    Full Text Available Abstract 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 fungus coated on test paper at a concentration of 3.3 × 106 conidia cm-2 reduced the daily survival of both mosquito species (HR = 3.14, p An. gambiae s.s relative to An. arabiensis (HR = 1.38, p 95% of mosquito cadavers in the treatment groups. The results indicate that M. anisopliae IP 46 has the potential to be a bio-control agent for African malaria vector species, and is a suitable candidate for further research and development.

  3. 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.

  4. 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

  5. Characterization of Plasmodium developmental transcriptomes in Anopheles gambiae midgut reveals novel regulators of malaria transmission.

    Science.gov (United States)

    Akinosoglou, Karolina A; Bushell, Ellen S C; Ukegbu, Chiamaka Valerie; Schlegelmilch, Timm; Cho, Jee-Sun; Redmond, Seth; Sala, Katarzyna; Christophides, George K; Vlachou, Dina

    2015-02-01

    The passage through the mosquito is a major bottleneck for malaria parasite populations and a target of interventions aiming to block disease transmission. Here, we used DNA microarrays to profile the developmental transcriptomes of the rodent malaria parasite Plasmodium berghei in vivo, in the midgut of Anopheles gambiae mosquitoes, from parasite stages in the midgut blood bolus to sporulating oocysts on the basal gut wall. Data analysis identified several distinct transcriptional programmes encompassing genes putatively involved in developmental processes or in interactions with the mosquito. At least two of these programmes are associated with the ookinete development that is linked to mosquito midgut invasion and establishment of infection. Targeted disruption by homologous recombination of two of these genes resulted in mutant parasites exhibiting notable infection phenotypes. GAMER encodes a short polypeptide with granular localization in the gametocyte cytoplasm and shows a highly penetrant loss-of-function phenotype manifested as greatly reduced ookinete numbers, linked to impaired male gamete release. HADO encodes a putative magnesium phosphatase with distinctive cortical localization along the concave ookinete periphery. Disruption of HADO compromises ookinete development leading to significant reduction of oocyst numbers. Our data provide important insights into the molecular framework underpinning Plasmodium development in the mosquito and identifies two genes with important functions at initial stages of parasite development in the mosquito midgut.

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

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    van Loon Joop JA

    2011-02-01

    Full Text Available Abstract 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 addition, volatiles produced by human skin bacteria are attractive to this mosquito species. The purpose of the current study was to test the effect of ten compounds present in the headspace of human bacteria on the host-seeking process of A. gambiae. The effect of each of the ten compounds on the attractiveness of a basic blend of ammonia, lactic and tetradecanoic acid to A. gambiae was examined. Methods The host-seeking response of A. gambiae was evaluated in a laboratory set-up using a dual-port olfactometer and in a semi-field facility in Kenya using MM-X traps. Odorants were released from LDPE sachets and placed inside the olfactometer as well as in the MM-X traps. Carbon dioxide was added in the semi-field experiments, provided from pressurized cylinders or fermenting yeast. Results The olfactometer and semi-field set-up allowed for high-throughput testing of the compounds in blends and in multiple concentrations. Compounds with an attractive or inhibitory effect were identified in both bioassays. 3-Methyl-1-butanol was the best attractant in both set-ups and increased the attractiveness of the basic blend up to three times. 2-Phenylethanol reduced the attractiveness of the basic blend in both bioassays by more than 50%. Conclusions Identification of volatiles released by human skin bacteria led to the discovery of compounds that have an impact on the host-seeking behaviour of A. gambiae. 3-Methyl-1-butanol may be used to increase mosquito trap catches, whereas 2-phenylethanol has potential as a spatial repellent. These two compounds could be applied in push-pull strategies to reduce mosquito numbers in malaria endemic areas.

  7. The toxicity of angiotensin converting enzyme inhibitors to larvae of the disease vectors Aedes aegypti and Anopheles gambiae.

    Science.gov (United States)

    Abu Hasan, Zatul-'Iffah; Williams, Helen; Ismail, Nur M; Othman, Hidayatulfathi; Cozier, Gyles E; Acharya, K Ravi; Isaac, R Elwyn

    2017-03-27

    The control of mosquitoes is threatened by the appearance of insecticide resistance and therefore new control chemicals are urgently required. Here we show that inhibitors of mosquito peptidyl dipeptidase, a peptidase related to mammalian angiotensin-converting enzyme (ACE), are insecticidal to larvae of the mosquitoes, Aedes aegypti and Anopheles gambiae. ACE inhibitors (captopril, fosinopril and fosinoprilat) and two peptides (trypsin-modulating oostatic factor/TMOF and a bradykinin-potentiating peptide, BPP-12b) were all inhibitors of the larval ACE activity of both mosquitoes. Two inhibitors, captopril and fosinopril (a pro-drug ester of fosinoprilat), were tested for larvicidal activity. Within 24 h captopril had killed >90% of the early instars of both species with 3(rd) instars showing greater resistance. Mortality was also high within 24 h of exposure of 1(st), 2(nd) and 3(rd) instars of An. gambiae to fosinopril. Fosinopril was also toxic to Ae. aegypti larvae, although the 1(st) instars appeared to be less susceptible to this pro-drug even after 72 h exposure. Homology models of the larval An. gambiae ACE proteins (AnoACE2 and AnoACE3) reveal structural differences compared to human ACE, suggesting that structure-based drug design offers a fruitful approach to the development of selective inhibitors of mosquito ACE enzymes as novel larvicides.

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

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    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.

  9. 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

  10. 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

  11. 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.

  12. 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.

  13. 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

  14. Larval density dependence in Anopheles gambiae s.s., the major African vector of malaria

    Science.gov (United States)

    Muriu, Simon M.; Coulson, Tim; Mbogo, Charles M.; Godfray, H. Charles J.

    2017-01-01

    Summary Anopheles gambiae sensu stricto is the most important vector of malaria in Africa although relatively little is known about the density-dependent processes determining its population size.Mosquito larval density was manipulated under semi-natural conditions using artificial larval breeding sites placed in the field in coastal Kenya; two experiments were conducted: one manipulating the density of a single cohort of larvae across a range of densities and the other employing fewer densities but with the treatments crossed with four treatments manipulating predator access.In the first experiment, larval survival, development rate and the size of the adult mosquito all decreased with larval density (controlling for block effects between 23% and 31% of the variance in the data could be explained by density).In the second experiment, the effects of predator manipulation were not significant, but again we observed strong density dependence in larval survival (explaining 30% of the variance).The results are compared with laboratory studies of A. gambiae larval competition and the few other studies conducted in the field, and the consequences for malaria control are discussed PMID:23163565

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

    Science.gov (United States)

    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.

  16. 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 ...

  17. 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

  18. Antiviral immunity of Anopheles gambiae is highly compartmentalized, with distinct roles for RNA interference and gut microbiota.

    Science.gov (United States)

    Carissimo, Guillaume; Pondeville, Emilie; McFarlane, Melanie; Dietrich, Isabelle; Mitri, Christian; Bischoff, Emmanuel; Antoniewski, Christophe; Bourgouin, Catherine; Failloux, Anna-Bella; Kohl, Alain; Vernick, Kenneth D

    2015-01-13

    Arboviruses are transmitted by mosquitoes and other arthropods to humans and animals. The risk associated with these viruses is increasing worldwide, including new emergence in Europe and the Americas. Anopheline mosquitoes are vectors of human malaria but are believed to transmit one known arbovirus, o'nyong-nyong virus, whereas Aedes mosquitoes transmit many. Anopheles interactions with viruses have been little studied, and the initial antiviral response in the midgut has not been examined. Here, we determine the antiviral immune pathways of the Anopheles gambiae midgut, the initial site of viral infection after an infective blood meal. We compare them with the responses of the post-midgut systemic compartment, which is the site of the subsequent disseminated viral infection. Normal viral infection of the midgut requires bacterial flora and is inhibited by the activities of immune deficiency (Imd), JAK/STAT, and Leu-rich repeat immune factors. We show that the exogenous siRNA pathway, thought of as the canonical mosquito antiviral pathway, plays no detectable role in antiviral defense in the midgut but only protects later in the systemic compartment. These results alter the prevailing antiviral paradigm by describing distinct protective mechanisms in different body compartments and infection stages. Importantly, the presence of the midgut bacterial flora is required for full viral infectivity to Anopheles, in contrast to malaria infection, where the presence of the midgut bacterial flora is required for protection against infection. Thus, the enteric flora controls a reciprocal protection tradeoff in the vector for resistance to different human pathogens.

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

    Science.gov (United States)

    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.

  20. 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

  1. 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.

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

    Science.gov (United States)

    Engdahl, Cecilia; Knutsson, Sofie; Fredriksson, Sten-Åke; Linusson, Anna; Bucht, Göran; Ekström, Fredrik

    2015-01-01

    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.

  3. 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

  4. Essential oils enhance the toxicity of permethrin against Aedes aegypti and Anopheles gambiae.

    Science.gov (United States)

    Gross, A D; Norris, E J; Kimber, M J; Bartholomay, L C; Coats, J R

    2017-03-01

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

  5. 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.

  6. Field Responses of Anopheles gambiae Complex (Diptera: Culicidae) in Liberia using Yeast-Generated Carbon Dioxide and Synthetic Lure-Baited Light Traps

    Science.gov (United States)

    2013-07-01

    VECTOR-BORNE DISEASES, SURVEILLANCE, PREVENTION Field Responses of Anopheles gambiae Complex (Diptera: Culicidae) in Liberia using Yeast-Generated...and light, bed-net, tent, andodor-baited traps (Mboera 2005). TheCenters forDiseaseControl and Prevention (CDC) light trap with its typical 4Ð6 W...using paper- clips. Although primarily developed and used to attract day ßying Stegomyia ( Aedes ) mosquitoes, blends of this lureÕs primary ingredients

  7. Inferring selection in the Anopheles gambiae species complex: an example from immune-related serine protease inhibitors

    Directory of Open Access Journals (Sweden)

    Little Tom J

    2009-06-01

    Full Text Available Abstract Background Mosquitoes of the Anopheles gambiae species complex are the primary vectors of human malaria in sub-Saharan Africa. Many host genes have been shown to affect Plasmodium development in the mosquito, and so are expected to engage in an evolutionary arms race with the pathogen. However, there is little conclusive evidence that any of these mosquito genes evolve rapidly, or show other signatures of adaptive evolution. Methods Three serine protease inhibitors have previously been identified as candidate immune system genes mediating mosquito-Plasmodium interaction, and serine protease inhibitors have been identified as hot-spots of adaptive evolution in other taxa. Population-genetic tests for selection, including a recent multi-gene extension of the McDonald-Kreitman test, were applied to 16 serine protease inhibitors and 16 other genes sampled from the An. gambiae species complex in both East and West Africa. Results Serine protease inhibitors were found to show a marginally significant trend towards higher levels of amino acid diversity than other genes, and display extensive genetic structuring associated with the 2La chromosomal inversion. However, although serpins are candidate targets for strong parasite-mediated selection, no evidence was found for rapid adaptive evolution in these genes. Conclusion It is well known that phylogenetic and population history in the An. gambiae complex can present special problems for the application of standard population-genetic tests for selection, and this may explain the failure of this study to detect selection acting on serine protease inhibitors. The pitfalls of uncritically applying these tests in this species complex are highlighted, and the future prospects for detecting selection acting on the An. gambiae genome are discussed.

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

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    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.

  9. 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.

  10. 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.

  11. 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.

  12. 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 mutaciones de resistencia knockdown (kdr) en Anopheles gambiae s.l. en Tanzania. Métodos Se recolectaron mosquitos Anopheles intradomiciliarios de 10 lugares diferentes y se evaluaron en busca de resistencia a insecticidas utilizando el protocolo estándar de la OMS. Mediante un diagnóstico molecular basado en la PCR se genotiparon los mosquitos y se detectaron los genotipos kdr. Resultados Los An. gambiae evaluados eran resistentes a 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

  13. Anopheles gambiae P450 reductase is highly expressed in oenocytes and in vivo knockdown increases permethrin susceptibility.

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    Lycett, G J; McLaughlin, L A; Ranson, H; Hemingway, J; Kafatos, F C; Loukeris, T G; Paine, M J I

    2006-06-01

    We describe an in vivo model for investigation of detoxification mechanisms of the mosquito Anopheles gambiae, important for the development of malaria control programmes. Cytochrome P450s are involved in metabolic insecticide resistance and require NADPH cytochrome P450 reductase (CPR) to function. Here we demonstrate that the major sites of adult mosquito CPR expression are oenocytes, mid-gut epithelia and head appendages. High CPR expression was also evident in Drosophila oenocytes indicating a general functional role in these insect cells. RNAi mediated knockdown drastically reduced CPR expression in oenocytes, and to a lesser extent in mid-gut epithelia; the head was unaffected. These flies showed enhanced sensitivity to permethrin, demonstrating a key role for abdominal/mid-gut P450s in pyrethroid metabolism, aiding the development of insecticides.

  14. 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.

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

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    Simões, Maria L; Gonçalves, Luzia; Silveira, Henrique

    2015-01-01

    Background Malaria is a worldwide infectious disease caused by Plasmodium parasites and transmitted by female Anopheles mosquitoes. The malaria vector mosquito Anopheles can trigger effective mechanisms to control completion of the Plasmodium lifecycle; the mosquito immune response to the parasite involves several pathways which are not yet well characterized. Plasmodium metabolite hemozoin has emerged as a potent immunostimulator of mammalian tissues. In this study, we aim to investigate the...

  16. Anopheles plumbeus (Diptera: Culicidae in Europe: a mere nuisance mosquito or potential malaria vector?

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    Schaffner Francis

    2012-11-01

    Full Text Available Abstract Background Anopheles plumbeus has been recognized as a minor vector for human malaria in Europe since the beginning of the 20th century. In recent years this tree hole breeding mosquito species appears to have exploited novel breeding sites, including large and organically rich man-made containers, with consequently larger mosquito populations in close vicinity to humans. This lead to investigate whether current populations of An. plumbeus would be able to efficiently transmit Plasmodium falciparum, the parasite responsible for the most deadly form of malaria. Methods Anopheles plumbeus immatures were collected from a liquid manure pit in Switzerland and transferred as adults to the CEPIA (Institut Pasteur, France where they were fed on P. falciparum gametocytes produced in vitro. Anopheles gambiae mosquitoes served as controls. Development of P. falciparum in both mosquito species was followed by microscopical detection of oocysts on mosquito midguts and by sporozoite detection in the head/thorax by PCR and microscopy. Results A total of 293 wild An. plumbeus females from four independent collections successfully fed through a membrane on blood containing P. falciparum gametocytes. Oocysts were observed in mosquito midguts and P. falciparum DNA was detected in head-thorax samples in all four experiments, demonstrating, on a large mosquito sample, that An. plumbeus is indeed receptive to P. falciparum NF54 and able to produce sporozoites. Importantly, the proportion of sporozoites-infected An. plumbeus was almost similar to that of An. gambiae (31 to 88% An. plumbeus versus 67 to 97% An. gambiae. However, the number of sporozoites produced was significantly lower in infected An. plumbeus. Conclusion The results show that a sample of field-caught An. plumbeus has a moderate to high receptivity towards P. falciparum. Considering the increased mobility of humans between Europe and malaria endemic countries and changes in environment and

  17. 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

  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. 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

  20. 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

  1. Transgenic Anopheles gambiae expressing an antimalarial peptide suffer no significant fitness cost.

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    McArthur, Clare C; Meredith, Janet M; Eggleston, Paul

    2014-01-01

    Mosquito-borne diseases present some of the greatest health challenges faced by the world today. In many cases, existing control measures are compromised by insecticide resistance, pathogen tolerance to drugs and the lack of effective vaccines. In light of these difficulties, new genetic tools for disease control programmes, based on the deployment of genetically modified mosquitoes, are seen as having great promise. Transgenic strains may be used to control disease transmission either by suppressing vector populations or by replacing susceptible with refractory genotypes. In practice, the fitness of the transgenic strain relative to natural mosquitoes will be a critical determinant of success. We previously described a transgenic strain of Anopheles gambiae expressing the Vida3 peptide into the female midgut following a blood-meal, which exhibited significant protection against malaria parasites. Here, we investigated the fitness of this strain relative to non-transgenic controls through comparisons of various life history traits. Experiments were designed, as far as possible, to equalize genetic backgrounds and heterogeneity such that fitness comparisons focussed on the presence and expression of the transgene cassette. We also employed reciprocal crosses to identify any fitness disturbance associated with inheritance of the transgene from either the male or female parent. We found no evidence that the presence or expression of the effector transgene or associated fluorescence markers caused any significant fitness cost in relation to larval mortality, pupal sex ratio, fecundity, hatch rate or longevity of blood-fed females. In fact, fecundity was increased in transgenic strains. We did, however, observe some fitness disturbances associated with the route of inheritance of the transgene. Maternal inheritance delayed male pupation whilst paternal inheritance increased adult longevity for both males and unfed females. Overall, in comparison to controls, there was

  2. Transgenic Anopheles gambiae expressing an antimalarial peptide suffer no significant fitness cost.

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    Clare C McArthur

    Full Text Available Mosquito-borne diseases present some of the greatest health challenges faced by the world today. In many cases, existing control measures are compromised by insecticide resistance, pathogen tolerance to drugs and the lack of effective vaccines. In light of these difficulties, new genetic tools for disease control programmes, based on the deployment of genetically modified mosquitoes, are seen as having great promise. Transgenic strains may be used to control disease transmission either by suppressing vector populations or by replacing susceptible with refractory genotypes. In practice, the fitness of the transgenic strain relative to natural mosquitoes will be a critical determinant of success. We previously described a transgenic strain of Anopheles gambiae expressing the Vida3 peptide into the female midgut following a blood-meal, which exhibited significant protection against malaria parasites. Here, we investigated the fitness of this strain relative to non-transgenic controls through comparisons of various life history traits. Experiments were designed, as far as possible, to equalize genetic backgrounds and heterogeneity such that fitness comparisons focussed on the presence and expression of the transgene cassette. We also employed reciprocal crosses to identify any fitness disturbance associated with inheritance of the transgene from either the male or female parent. We found no evidence that the presence or expression of the effector transgene or associated fluorescence markers caused any significant fitness cost in relation to larval mortality, pupal sex ratio, fecundity, hatch rate or longevity of blood-fed females. In fact, fecundity was increased in transgenic strains. We did, however, observe some fitness disturbances associated with the route of inheritance of the transgene. Maternal inheritance delayed male pupation whilst paternal inheritance increased adult longevity for both males and unfed females. Overall, in comparison to

  3. Attractiveness of MM-X traps baited with human or synthetic odor to mosquitoes (Diptera: Culicidae) in The Gambia.

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    Qiu, Yu Tong; Smallegange, Renate C; Ter, Braak Cajo J F; Spitzen, Jeroen; Van Loon, Joop J A; Jawara, Musa; Milligan, Paul; Galimard, Agnes M; Van Beek, Teris A; Knols, Bart G J; Takken, Willem

    2007-11-01

    Chemical cues play an important role in the host-seeking behavior of blood-feeding mosquitoes (Diptera: Culicidae). A field study was carried out in The Gambia to investigate the effects of human odor or synthetic odor blends on the attraction of mosquitoes. MM-X traps baited with 16 odor blends to which carbon dioxide (CO2) was added were tested in four sets of experiments. In a second series of experiments, MM-X traps with 14 odor blends without CO2 were tested. A blend of ammonia and L-lactic acid with or without CO2 was used as control odor in series 1 and 2, respectively. Centers for Disease Control and Prevention (CDC) traps were placed in a traditional house and an experimental house to monitor mosquito densities during the experiments. The MM-X traps caught a total number of 196,756 mosquitoes, with the most abundant species belonging to the genera Mansonia (70.6%), Anopheles (17.5%), and Culex (11.5%). The most abundant mosquito species caught by the CDC traps (56,290 in total) belonged to the genera Mansonia (59.4%), Anopheles (16.0% An. gambiae s.l. Giles, and 11.3% An. ziemanni Grünberg), and Culex (11.6%). MM-X traps baited with synthetic blends were in many cases more attractive than MM-X traps baited with human odors. Addition of CO2 to synthetic odors substantially increased the catch of all mosquito species in the MM-X traps. A blend of ammonia + L-lactic acid + CO, + 3-methylbutanoic acid was the most attractive odor for most mosquito species. The candidate odor blend shows the potential to enhance trap collections so that traps will provide better surveillance and possible control.

  4. Genetic variation of male reproductive success in a laboratory population of Anopheles gambiae

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    Voordouw Maarten J

    2007-07-01

    Full Text Available Abstract Background For Anopheline mosquitoes, the vectors of human malaria, genetic variation in male reproductive success can have important consequences for any control strategy based on the release of transgenic or sterile males. Methods A quantitative genetics approach was used to test whether there was a genetic component to variation in male reproductive success in a laboratory population of Anopheles gambiae. Swarms of full sibling brothers were mated with a fixed number of females and their reproductive success was measured as (1 proportion of ovipositing females, (2 proportion of ovipositing females that produced larvae, (3 proportion of females that produced larvae, (4 number of eggs laid per female, (5 number of larvae per ovipositing female and (6 number of larvae per female. Results The proportion of ovipositing females (trait 1 and the proportion of ovipositing females that produced larvae (trait 2 differed among full sib families, suggesting a genetic basis of mating success. In contrast, the other measures of male reproductive success showed little variation due to the full sib families, as their variation are probably mostly due to differences among females. While age at emergence and wing length of the males were also heritable, they were not associated with reproductive success. Larger females produced more eggs, but males did not prefer such partners. Conclusion The first study to quantify genetic variation for male reproductive success in A. gambiae found that while the initial stages of male reproduction (i.e. the proportion of ovipositing females and the proportion of ovipositing females that produced larvae had a genetic basis, the overall reproductive success (i.e. the mean number of larvae per female did not.

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

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    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

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

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    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.

  7. 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.

  8. Egg hatching, larval movement and larval survival of the malaria vector Anopheles gambiae in desiccating habitats

    NARCIS (Netherlands)

    Koenraadt, C.J.M.; Paaijmans, K.P.; Githeko, A.K.; Knols, B.G.J.; Takken, W.

    2003-01-01

    Background - Although the effects of rainfall on the population dynamics of the malaria vector Anopheles gambiae have been studied in great detail, the effects of dry periods on its survival remain less clear. Methods - The effects of drying conditions were simulated by creating desiccated habitats,

  9. Effectiveness of synthetic versus natural human volatiles as attractants for Anopheles gambiae (Diptera: Culicidae) sensu stricto

    NARCIS (Netherlands)

    Smallegange, R.C.; Knols, B.G.J.; Takken, W.

    2010-01-01

    Females of the African malaria vector, Anopheles gambiae Giles sensu stricto, use human volatiles to find their blood-host. Previous work has shown that ammonia, lactic acid, and aliphatic carboxylic acids significantly affect host orientation and attraction of this species, In the current study, th

  10. 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

  11. 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

  12. 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.

  13. 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

  14. 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.

  15. Site-specific genetic engineering of the Anopheles gambiae Y chromosome.

    Science.gov (United States)

    Bernardini, Federica; Galizi, Roberto; Menichelli, Miriam; Papathanos, Philippos-Aris; Dritsou, Vicky; Marois, Eric; Crisanti, Andrea; Windbichler, Nikolai

    2014-05-27

    Despite its function in sex determination and its role in driving genome evolution, the Y chromosome remains poorly understood in most species. Y chromosomes are gene-poor, repeat-rich and largely heterochromatic and therefore represent a difficult target for genetic engineering. The Y chromosome of the human malaria vector Anopheles gambiae appears to be involved in sex determination although very little is known about both its structure and function. Here, we characterize a transgenic strain of this mosquito species, obtained by transposon-mediated integration of a transgene construct onto the Y chromosome. Using meganuclease-induced homologous repair we introduce a site-specific recombination signal onto the Y chromosome and show that the resulting docking line can be used for secondary integration. To demonstrate its utility, we study the activity of a germ-line-specific promoter when located on the Y chromosome. We also show that Y-linked fluorescent transgenes allow automated sex separation of this important vector species, providing the means to generate large single-sex populations. Our findings will aid studies of sex chromosome function and enable the development of male-exclusive genetic traits for vector control.

  16. Biting behavior of Anopheles mosquitoes in Costa Marques, Rondonia, Brazil

    Directory of Open Access Journals (Sweden)

    Terry A. Klein

    1991-03-01

    Full Text Available Mosquito collections were made in and near Costa Marques, Rondonia, Brazil, to determine anopheline anthropophilic/zoophilic behavior. Collections from a non-illuminated, bovine-baited trap and indoor and outdoor human-bait collections were compared. Anopheles darlingi and Anopheles deaneorum were more anthropophilic than the other anophelines collected. The remainder of the Anopheles species were collected much morefrequently in bovine-baited traps than in human-bait collections. Anopheles darlingi and An. deaneorum were more frequently collected inside houses than the other anopheline species. But, when collections were made in a house with numerous openings in the walls, there were few differences in the percentages of each species biting man indoors versus outdoors. Anopheles darlingi was the predominant mosquito collected, both inside and outside houses, and had the strongest anthropophilic feeding behavior of the anophelines present.Para determinar o comportamento antropofilico e zoofilico dos anofelinos, foram capturados mosquitos na periferia e na zona urbana de Costa Marques, Rondônia, Brasil. Foram comparadas as capturas feitas à noite, com iscas bovinas e humanas, dentro efora de casa. O Anopheles darlingi e o Anopheles deaneorumforam mais antropojilicos do que os outros anofelinos capturados. O restante das espécies anofelinas foi capturado mais freqüentemente nas iscas bovinas do que nas humanas. Anopheles darlingi e Anopheles deaneorumforam capturados dentro de casa com mais freqüência do que as outras espécies anofelinas. Porém, quando a captura foi feita em casas com muitas aberturas nas paredes houve pouca diferença nas porcentagens de cada espécie sugadora de humanos dentro efora de casa. Anopheles darlingi foi o mosquito capturado com mais freqüência, dentro e fora de casa, e apresentava maior antropofilia em relação aos outros anofelinos presentes.

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

    Directory of Open Access Journals (Sweden)

    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.

  18. The Mode of Action of Spatial Repellents and Their Impact on Vectorial Capacity of Anopheles gambiae sensu stricto

    Science.gov (United States)

    Ogoma, Sheila B.; Ngonyani, Hassan; Simfukwe, Emmanuel T.; Mseka, Antony; Moore, Jason; Maia, Marta F.; Moore, Sarah J.; Lorenz, Lena M.

    2014-01-01

    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. PMID:25485850

  19. 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.

  20. Insecticidal activities of bark, leaf and seed extracts of Zanthoxylum heitzii against the African malaria vector Anopheles gambiae.

    Science.gov (United States)

    Overgaard, Hans J; Sirisopa, Patcharawan; Mikolo, Bertin; Malterud, Karl E; Wangensteen, Helle; Zou, Yuan-Feng; Paulsen, Berit S; Massamba, Daniel; Duchon, Stephane; Corbel, Vincent; Chandre, Fabrice

    2014-12-17

    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.

  1. 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.

  2. 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

  3. Targeting the X chromosome during spermatogenesis induces Y chromosome transmission ratio distortion and early dominant embryo lethality in Anopheles gambiae.

    OpenAIRE

    Nikolai Windbichler; Philippos Aris Papathanos; Andrea Crisanti

    2008-01-01

    Author Summary A. gambiae mosquitoes are the main vectors of human malaria. The inadequacy of existing control measures for these mosquitoes has prompted research into methods for genetic control. We have genetically engineered A. gambiae mosquitoes to express, during spermatozoa development, an enzyme that selectively cuts a DNA sequence present only on a family of essential genes located on the X chromosome. We found that in heterozygous male mosquitoes, this genetic modification induced co...

  4. 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

  5. Performance of five food regimes on Anopheles gambiae senso stricto larval rearing to adult emergence in insectary.

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    Happiness S Kivuyo

    Full Text Available BACKGROUND: Rearing of Anopheles gambiae s.s mosquitoes in insectary with quality cheap food sources is of paramount importance for better and healthy colony. This study evaluated larval survival and the development rate of aquatic stages of An.gambiae s.s under five food regimes; tetramin fish food (a standard insectary larval food, maize pollen, Cerelac, green filamentous algae and dry powdered filamentous algae. METHODS: Food materials were obtained from different sources, cerelac was made locally, fresh filamentous algae was taken from water bodies, dry filamentous algae was ground to powder after it was dried under shade, and maize pollen was collected from the flowering maize. Each food source type was used to feed three densities of mosquito larvae 20, 60, and 100 in six replicates each. Larval age structure was monitored daily until pupation and subsequently adult emergence. Tetramin was used and taken as a standard food source for An. gambiae s.s. larvae feeding in Insectary. RESULTS: Larval survivorship using maize pollen and Tetramin fish food was statistically insignificant (P = 0.564. However when compared to other food regime survivorship was significantly different with Tetramin fish food performing better than cerelac (P<0.001, dry algae (P<0.001 and fresh algae (P<0.001. The pupation rates and sex ratio of emerging adults had significant differences among the food regimes. CONCLUSION: The findings of this study have shown that maize pollen had closely similar nutritional value for larval survivorship to tetramin fish food, a standard larvae food in insectary. Further studies are required to assess the effect of food sources on various life traits of the emerged adults.

  6. 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

  7. Larval habitats of Anopheles gambiae s.s. (Diptera: Culicidae influences vector competence to Plasmodium falciparum parasites

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    Gouagna Louis C

    2007-04-01

    Full Text Available Abstract Background The origin of highly competent malaria vectors has been linked to productive larval habitats in the field, but there isn't solid quantitative or qualitative data to support it. To test this, the effect of larval habitat soil substrates on larval development time, pupation rates and vector competence of Anopheles gambiae to Plasmodium falciparum were examined. Methods Soils were collected from active larval habitats with sandy and clay substrates from field sites and their total organic matter estimated. An. gambiae larvae were reared on these soil substrates and the larval development time and pupation rates monitored. The emerging adult mosquitoes were then artificially fed blood with infectious P. falciparum gametocytes from human volunteers and their midguts examined for oocyst infection after seven days. The wing sizes of the mosquitoes were also measured. The effect of autoclaving the soil substrates was also evaluated. Results The total organic matter was significantly different between clay and sandy soils after autoclaving (P = 0.022. A generalized liner model (GLM analysis identified habitat type (clay soil, sandy soil, or lake water and autoclaving (that reduces presence of microbes as significant factors affecting larval development time and oocyst infection intensities in adults. Autoclaving the soils resulted in the production of significantly smaller sized mosquitoes (P = 0.008. Autoclaving clay soils resulted in a significant reduction in Plasmodium falciparum oocyst intensities (P = 0.041 in clay soils (unautoclaved clay soils (4.28 ± 0.18 oocysts/midgut; autoclaved clay soils = 1.17 ± 0.55 oocysts/midgut although no difference (P = 0.480 in infection rates was observed between clay soils (10.4%, sandy soils (5.3% or lake water (7.9%. Conclusion This study suggests an important nutritional role for organic matter and microbial fauna on mosquito fitness and vector competence. It shows that the quality of

  8. [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.

  9. Choice of resting sites by Anopheles gambiae (Diptera: Culici) in Mwea Rice Irrigation Scheme, Kirinyaga District, Kenya.

    Science.gov (United States)

    Mutinga, M J; Odhiambo, T R; Kamau, C C; Odulaja, A; Amimo, F A; Wachira, D W

    1995-03-01

    Investigations on Anopheles gambiae were carried out in Mwea Rice Irrigation Scheme, Kenya, to determine their preference for resting on various colours of fabrics and bare walls inside rural houses. The inside wall surface of each house was divided into upper, middle and lower sections and the section further partitioned into subsections measuring 0.6 x 0.3m. Two sets of experiments were conducted: one in which the walls were partly fitted with one colour of fabric at a time (double-choice situation), while in the other the walls were fitted with a combination of six colours (red, black, yellow, white, green and blue) of fabric at once, (multiple-choice situation). The number of mosquitoes resting on each of the pieces of cloth pinned to the walls and the uncovered subsections of the wall were recorded for each section of the wall after 24 hours. The results showed resting preference for cloth covered parts of the walls. The white colour was found to best attract mosquitoes followed by colours red, yellow, black, blue and green. The hanging strips in the middle of the houses had fewer mosquitoes resting on them than those on the wall.

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

    Directory of Open Access Journals (Sweden)

    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.

  11. Identification and validation of a gene causing cross-resistance between insecticide classes in Anopheles gambiae from Ghana.

    Science.gov (United States)

    Mitchell, Sara N; Stevenson, Bradley J; Müller, Pie; Wilding, Craig S; Egyir-Yawson, Alexander; Field, Stuart G; Hemingway, Janet; Paine, Mark J I; Ranson, Hilary; Donnelly, Martin James

    2012-04-17

    In the last decade there have been marked reductions in malaria incidence in sub-Saharan Africa. Sustaining these reductions will rely upon insecticides to control the mosquito malaria vectors. We report that in the primary African malaria vector, Anopheles gambiae sensu stricto, a single enzyme, CYP6M2, confers resistance to two classes of insecticide. This is unique evidence in a disease vector of cross-resistance associated with a single metabolic gene that simultaneously reduces the efficacy of two of the four classes of insecticide routinely used for malaria control. The gene-expression profile of a highly DDT-resistant population of A. gambiae s.s. from Ghana was characterized using a unique whole-genome microarray. A number of genes were significantly overexpressed compared with two susceptible West African colonies, including genes from metabolic families previously linked to insecticide resistance. One of the most significantly overexpressed probe groups (false-discovery rate-adjusted P metabolize both type I and type II pyrethroids in recombinant protein assays. Using in vitro assays we show that recombinant CYP6M2 is also capable of metabolizing the organochlorine insecticide DDT in the presence of solubilizing factor sodium cholate.

  12. Metabolic and Target-Site Mechanisms Combine to Confer Strong DDT Resistance in Anopheles gambiae

    OpenAIRE

    2014-01-01

    The development of resistance to insecticides has become a classic exemplar of evolution occurring within human time scales. In this study we demonstrate how resistance to DDT in the major African malaria vector Anopheles gambiae is a result of both target-site resistance mechanisms that have introgressed between incipient species (the M- and S-molecular forms) and allelic variants in a DDT-detoxifying enzyme. Sequencing of the detoxification enzyme, Gste2, from DDT resistant and susceptible ...

  13. 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.

  14. 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

  15. Evaluation of PermaNet 3.0 a deltamethrin-PBO combination net against Anopheles gambiae and pyrethroid resistant Culex quinquefasciatus mosquitoes: an experimental hut trial in Tanzania

    OpenAIRE

    Malima Robert; Maxwell Caroline; Magesa Stephen; Tungu Patrick; Masue Dennis; Sudi Wema; Myamba Joseph; Pigeon Olivier; Rowland Mark

    2010-01-01

    Abstract Background Combination mosquito nets incorporating two unrelated insecticides or insecticide plus synergist are designed to control insecticide resistant mosquitoes. PermaNet 3.0 is a long-lasting combination net incorporating deltamethrin on the side panels and a mixture of deltamethrin and synergist piperonyl butoxide (PBO) on the top panel. PBO is an inhibitor of mixed function oxidases implicated in pyrethroid resistance. Method An experimental hut trial comparing PermaNet 3.0, P...

  16. 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

  17. 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.

  18. Studies on the bionomics of male Anopheles gambiae Giles and male Anopheles funestus Giles from southern Mozambique.

    Science.gov (United States)

    Charlwood, J D

    2011-12-01

    Little is known about the fitness of wild male mosquitoes, the females of which are vectors of malaria. The problem of studying male biology has been exacerbated by difficulties associated with catching them. In southern Mozambique, however, almost the entire adult population of An. funestus and An. gambiae s.l. rest inside houses. They leave in a dusk exodus, which makes them easy to collect. In 8,348 exit collections from a village from 2003 to 2009, 567,195 male An. funestus and 34,591 male An. gambiae s.l. were collected. During the study, numbers of An. funestus increased but numbers of An. gambiae s.l. declined to the point of extinction. Overall numbers of An. gambiae s.l. were positively correlated with temperature, whilst the relationship between temperature and numbers of An. funestus changed from an initially positive one in the first three years of the study to a negative one in the last three years. Marked males were recaptured up to 300 m from the release site, with most recaptures occurring within 150 m. Estimated mean daily survival of male An. funestus was 0.86 (95% C.I. 0.869-0.850). For the years 2003-2007, estimated mean daily survival of male An gambiae s.l. was 0.660 (95% C.I. 0.682-0.638). For either species, there was no relationship between mean weekly temperature and estimated daily survival. These results imply that males of An. funestus live as long as females but have a relatively short flight range. They are discussed in the light of possible release strategies of sterile or genetically modified mosquitoes.

  19. 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

  20. Genetic and environmental factors associated with laboratory rearing affect survival and assortative mating but not overall mating success in Anopheles gambiae sensu stricto.

    Science.gov (United States)

    Paton, Doug; Touré, Mahamoudou; Sacko, Adama; Coulibaly, Mamadou B; Traoré, Sékou F; Tripet, Frédéric

    2013-01-01

    Anopheles gambiae sensu stricto, the main vector of malaria in Africa, is characterized by its vast geographical range and complex population structure. Assortative mating amongst the reproductively isolated cryptic forms that co-occur in many areas poses unique challenges for programs aiming to decrease malaria incidence via the release of sterile or genetically-modified mosquitoes. Importantly, whether laboratory-rearing affects the ability of An. gambiae individuals of a given cryptic taxa to successfully mate with individuals of their own form in field conditions is still unknown and yet crucial for mosquito-releases. Here, the independent effects of genetic and environmental factors associated with laboratory rearing on male and female survival, mating success and assortative mating were evaluated in the Mopti form of An. gambiae over 2010 and 2011. In semi-field enclosures experiments and despite strong variation between years, the overall survival and mating success of male and female progeny from a laboratory strain was not found to be significantly lower than those of the progeny of field females from the same population. Adult progeny from field-caught females reared at the larval stage in the laboratory and from laboratory females reared outdoors exhibited a significant decrease in survival but not in mating success. Importantly, laboratory individuals reared as larvae indoors were unable to mate assortatively as adults, whilst field progeny reared either outdoors or in the laboratory, as well as laboratory progeny reared outdoors all mated significantly assortatively. These results highlight the importance of genetic and environment interactions for the development of An. gambiae's full mating behavioral repertoire and the challenges this creates for mosquito rearing and release-based control strategies.

  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. 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. 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.

  4. Anopheles (Anopheles) petragnani Del Vecchio 1939-a new mosquito species for Germany.

    Science.gov (United States)

    Becker, Norbert; Pfitzner, Wolf Peter; Czajka, Christina; Kaiser, Achim; Weitzel, Thomas

    2016-07-01

    The so far known species of the Anopheles Claviger Complex, Anopheles claviger s.s. and Anopheles petragnani, can only be distinguished by partial overlapping characteristics of immature stages and by nucleotide sequence variation of the genomic ribosomal DNA (rDNA) internal transcribed spacer 2 (ITS2) region. The known distribution of An. petragnani is so far restricted to the western Mediterranean region, whereas An. claviger s.s. occurs across most of Europe, up to the Middle East and North Africa. In our study, we investigated the larval mosquito fauna in rock pools of the Murg valley (Black Forest, Germany) once a month from April to December 2015.Among other species, larvae belonging to the Anopheles Claviger Complex were found. The fourth instar larvae were morphologically identified by chaetotaxy of the head and abdomen. The results were confirmed by a multiplex PCR and additional sequencing of the amplificates.Of the 1289 collected larvae from the rock pools, seven belonged to the Anopheles Claviger Complex. Five individuals were determined morphologically as An. petragnani and two as An. claviger s.s. The associated mosquito fauna comprised of Aedes japonicus japonicus (548 individuals), Culex pipiens s.l. and Culex torrentium (493 individuals) and Culex hortensis (241 individuals).This is the first record of An. petragnani north of the Alps. Further studies will reveal whether this is an isolated population of An. petragnani and if the investigated rock pool breeding sites represent typical habitats of this species in temperate regions in Central Europe.

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

    Directory of Open Access Journals (Sweden)

    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

  6. The role of cow urine in the oviposition site preference of culicine and Anopheles mosquitoes

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    Kweka Eliningaya J

    2011-09-01

    Full Text Available 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 Oviposition response evaluation in laboratory conditions was carried out in mosquito rearing cages. The oviposition substrates were located in parallel or in diagonal positions inside the cage. Urine evaluation against gravid females of An. arabiensis and Cx. quinquefasciatus was carried out at Day 1, Day 3 and Day 7. Five millilitres (mls of cow urine was added to oviposition substrate while de-chlorinated water was used as a control. In field experiments, 500 mls of cow urine was added in artificial habitats with 2500 mls of de-chlorinated water and 2 kgs of soil. The experiment was monitored for thirty consecutive days, eggs were collected daily from the habitats at 7.00 hrs. Data analysis was performed using parametric and non-parametric tests for treatments and controls while attraction of the oviposition substrate in each species was presented using Oviposition Activity Index (OAI. Results The OAI was positive with ageing of cattle urine in culicine species in both laboratory and field experiments. The OAI for anopheline species was positive with fresh urine. The OAI during the rainy season was positive for all species tested while in the dry season the OAI for culicine spp and Anopheles gambiae s.l., changed with time from positive to negative values. Based on linear model analysis, seasons and treatments had a significant effect on the number of eggs laid in habitats, even though the number of days had no effect. Conclusion Oviposition substrates treated with

  7. 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...

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

    Directory of Open Access Journals (Sweden)

    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

  9. Association mapping of insecticide resistance in wild Anopheles gambiae populations: major variants identified in a low-linkage disequilbrium genome.

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    David Weetman

    Full Text Available BACKGROUND: Association studies are a promising way to uncover the genetic basis of complex traits in wild populations. Data on population stratification, linkage disequilibrium and distribution of variant effect-sizes for different trait-types are required to predict study success but are lacking for most taxa. We quantified and investigated the impacts of these key variables in a large-scale association study of a strongly selected trait of medical importance: pyrethroid resistance in the African malaria vector Anopheles gambiae. METHODOLOGY/PRINCIPAL FINDINGS: We genotyped ≈1500 resistance-phenotyped wild mosquitoes from Ghana and Cameroon using a 1536-SNP array enriched for candidate insecticide resistance gene SNPs. Three factors greatly impacted study power. (1 Population stratification, which was attributable to co-occurrence of molecular forms (M and S, and cryptic within-form stratification necessitating both a partitioned analysis and genomic control. (2 All SNPs of substantial effect (odds ratio, OR>2 were rare (minor allele frequency, MAF<0.05. (3 Linkage disequilibrium (LD was very low throughout most of the genome. Nevertheless, locally high LD, consistent with a recent selective sweep, and uniformly high ORs in each subsample facilitated significant direct and indirect detection of the known insecticide target site mutation kdr L1014F (OR≈6; P<10(-6, but with resistance level modified by local haplotypic background. CONCLUSION: Primarily as a result of very low LD in wild A. Gambiae, LD-based association mapping is challenging, but is feasible at least for major effect variants, especially where LD is enhanced by selective sweeps. Such variants will be of greatest importance for predictive diagnostic screening.

  10. 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

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

    NARCIS (Netherlands)

    Bukhari, 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 (specie

  12. Dynamics of multiple insecticide resistance in the malaria vector Anopheles gambiae in a rice growing area in South-Western Burkina Faso

    Directory of Open Access Journals (Sweden)

    Ouédraogo Jean-Bosco

    2008-09-01

    Full Text Available Abstract Background Insecticide resistance of the main malaria vector, Anopheles gambiae, has been reported in south-western Burkina Faso, West Africa. Cross-resistance to DDT and pyrethroids was conferred by alterations at site of action in the sodium channel, the Leu-Phe kdr mutation; resistance to organophosphates and carbamates resulted from a single point mutation in the oxyanion hole of the acetylcholinesterase enzyme designed as ace-1R. Methods An entomological survey was carried out during the rainy season of 2005 at Vallée du Kou, a rice growing area in south-western Burkina Faso. At the Vallée du Kou, both insecticide resistance mechanisms have been previously described in the M and S molecular forms of An. gambiae. This survey aimed i to update the temporal dynamics and the circumsporozoite infection rate of the two molecular forms M and S of An. gambiae ii to update the frequency of the Leu-Phe kdr mutation within these forms and finally iii to investigate the occurrence of the ace-1R mutation. Mosquitoes collected by indoor residual collection and by human landing catches were counted and morphologically identified. Species and molecular forms of An. gambiae, ace-1R and Leu-Phe kdr mutations were determined using PCR techniques. The presence of the circumsporozoite protein of Plasmodium falciparum was determined using ELISA. Results Anopheles gambiae populations were dominated by the M form. However the S form occurred in relative important proportion towards the end of the rainy season with a maximum peak in October at 51%. Sporozoite rates were similar in both forms. The frequency of the Leu-Phe kdr mutation in the S form reached a fixation level while it is still spreading in the M form. Furthermore, the ace-1R mutation prevailed predominately in the S form and has just started spreading in the M form. The two mutations occurred concomitantly both in M and S populations. Conclusion These results showed that the Vallée du Kou

  13. 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

  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. Metabolic and target-site mechanisms combine to confer strong DDT resistance in Anopheles gambiae.

    Science.gov (United States)

    Mitchell, Sara N; Rigden, Daniel J; Dowd, Andrew J; Lu, Fang; Wilding, Craig S; Weetman, David; Dadzie, Samuel; Jenkins, Adam M; Regna, Kimberly; Boko, Pelagie; Djogbenou, Luc; Muskavitch, Marc A T; Ranson, Hilary; Paine, Mark J I; Mayans, Olga; Donnelly, Martin J

    2014-01-01

    The development of resistance to insecticides has become a classic exemplar of evolution occurring within human time scales. In this study we demonstrate how resistance to DDT in the major African malaria vector Anopheles gambiae is a result of both target-site resistance mechanisms that have introgressed between incipient species (the M- and S-molecular forms) and allelic variants in a DDT-detoxifying enzyme. Sequencing of the detoxification enzyme, Gste2, from DDT resistant and susceptible strains of An. gambiae, revealed a non-synonymous polymorphism (I114T), proximal to the DDT binding domain, which segregated with strain phenotype. Recombinant protein expression and DDT metabolism analysis revealed that the proteins from the susceptible strain lost activity at higher DDT concentrations, characteristic of substrate inhibition. The effect of I114T on GSTE2 protein structure was explored through X-ray crystallography. The amino acid exchange in the DDT-resistant strain introduced a hydroxyl group nearby the hydrophobic DDT-binding region. The exchange does not result in structural alterations but is predicted to facilitate local dynamics and enzyme activity. Expression of both wild-type and 114T alleles the allele in Drosophila conferred an increase in DDT tolerance. The 114T mutation was significantly associated with DDT resistance in wild caught M-form populations and acts in concert with target-site mutations in the voltage gated sodium channel (Vgsc-1575Y and Vgsc-1014F) to confer extreme levels of DDT resistance in wild caught An. gambiae.

  17. 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.

  18. 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.

  19. Innexin AGAP001476 is critical for mediating anti-Plasmodium responses in Anopheles mosquitoes.

    Science.gov (United States)

    Li, Michelle W M; Wang, Jiuling; Zhao, Yang O; Fikrig, Erol

    2014-09-05

    The Toll and IMD pathways are known to be induced upon Plasmodium berghei and Plasmodium falciparum infection, respectively. It is unclear how Plasmodium or other pathogens in the blood meal and their invasion of the midgut epithelium would trigger the innate immune responses in immune cells, in particular hemocytes. Gap junctions, which can mediate both cell-to-cell and cell-to-extracellular communication, may participate in this signal transduction. This study examined whether innexins, gap junction proteins in insects, are involved in anti-Plasmodium responses in Anopheles gambiae. Inhibitor studies using carbenoxolone indicated that blocking innexons resulted in an increase in Plasmodium oocyst number and infection prevalence. This was accompanied by a decline in TEP1 levels in carbenoxolone-treated mosquitoes. Innexin AGAP001476 mRNA levels in midguts were induced during Plasmodium infection and a knockdown of AGAP001476, but not AGAP006241, caused an induction in oocyst number. Silencing AGAP001476 caused a concurrent increase in vitellogenin levels, a TEP1 inhibitor, in addition to a reduced level of TEP1-LRIM1-APL1C complex in hemolymph. Both vitellogenin and TEP1 are regulated by Cactus under the Toll pathway. Simultaneous knockdown of cactus and AGAP001476 failed to reverse the near refractoriness induced by the knockdown of cactus, suggesting that the AGAP001476-mediated anti-Plasmodium response is Cactus-dependent. These data demonstrate a critical role for innexin AGAP001476 in mediating innate immune responses against Plasmodium through Toll pathway in mosquitoes.

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

    Science.gov (United States)

    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.

  1. Combining indoor residual spraying with chlorfenapyr and long-lasting insecticidal bed nets for improved control of pyrethroid-resistant Anopheles gambiae: an experimental hut trial in Benin

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    Ngufor Corine

    2011-11-01

    Full Text Available Abstract Background Neither indoor residual spraying (IRS nor long-lasting insecticidal nets (LLINs are able to fully interrupt transmission in holoendemic Africa as single interventions. The combining of IRS and LLINs presents an opportunity for improved control and management of pyrethroid resistance through the simultaneous presentation of unrelated insecticides. Method Chlorfenapyr IRS and a pyrethroid-impregnated polyester LLIN (WHO approved were tested separately and together in experimental huts in southern Benin against pyrethroid resistant Anopheles gambiae and Culex quinquefasciatus. The bed nets were deliberately holed with either six or 80 holes to examine the effect of increasing wear and tear on protectiveness. Anopheles gambiae were genotyped for the kdr gene to assess the combination's potential to prevent the selection of pyrethroid resistance. Results The frequency of kdr was 84%. The overall mortality rates of An. gambiae were 37% and 49% with the six-hole and 80-hole LLINs, respectively, and reached 57% with chlorfenapyr IRS. Overall mortality rates were significantly higher with the combination treatments (82-83% than with the LLIN or IRS individual treatments. Blood feeding (mosquito biting rates were lowest with the 6-hole LLIN (12%, intermediate with the 80-hole LLIN (32% and highest with untreated nets (56% with the 6-hole and 54% with the 80-hole nets. Blood feeding (biting rates and repellency of mosquitoes with the combination of LLIN and chlorfenapyr IRS showed significant improvement compared to the IRS treatment but did not differ from the LLIN treatments indicating that the LLINs were the primary agents of personal protection. The combination killed significantly higher proportions of Cx. quinquefasciatus (51%, 41% than the LLIN (15%, 13% or IRS (32% treatments. Conclusion The chlorfenapyr IRS component was largely responsible for controlling pyrethroid-resistant mosquitoes and the LLIN component was largely

  2. The transmission potential of malaria-infected mosquitoes (An.gambiae-Keele, An.arabiensis-Ifakara) is altered by the vertebrate blood type they consume during parasite development

    Science.gov (United States)

    Emami, S. Noushin; Ranford-Cartwright, Lisa C.; Ferguson, Heather M.

    2017-01-01

    The efficiency of malaria parasite development within mosquito vectors (sporogony) is a critical determinant of transmission. Sporogony is thought to be controlled by environmental conditions and mosquito/parasite genetic factors, with minimal contribution from mosquito behaviour during the period of parasite development. We tested this assumption by investigating whether successful sporogony of Plasmodium falciparum parasites through to human-infectious transmission stages is influenced by the host species upon which infected mosquitoes feed. Studies were conducted on two major African vector species that generally are found to differ in their innate host preferences: Anopheles arabiensis and An. gambiae sensu stricto. We show that the proportion of vectors developing transmissible infections (sporozoites) was influenced by the source of host blood consumed during sporogony. The direction of this effect was associated with the innate host preference of vectors: higher sporozoite prevalences were generated in the usually human-specialist An. gambiae s.s. feeding on human compared to cow blood, whereas the more zoophilic An. arabiensis had significantly higher prevalences after feeding on cow blood. The potential epidemiological implications of these results are discussed. PMID:28094293

  3. Laser induced mortality of Anopheles stephensi mosquitoes

    Science.gov (United States)

    Keller, Matthew D.; Leahy, David J.; Norton, Bryan J.; Johanson, Threeric; Mullen, Emma R.; Marvit, Maclen; Makagon, Arty

    2016-02-01

    Small, flying insects continue to pose great risks to both human health and agricultural production throughout the world, so there remains a compelling need to develop new vector and pest control approaches. Here, we examined the use of short (mosquitoes, which were chosen as a representative species. The mortality of mosquitoes exposed to laser pulses of various wavelength, power, pulse duration, and spot size combinations was assessed 24 hours after exposure. For otherwise comparable conditions, green and far-infrared wavelengths were found to be more effective than near- and mid-infrared wavelengths. Pulses with larger laser spot sizes required lower lethal energy densities, or fluence, but more pulse energy than for smaller spot sizes with greater fluence. Pulse duration had to be reduced by several orders of magnitude to significantly lower the lethal pulse energy or fluence required. These results identified the most promising candidates for the lethal laser component in a system being designed to identify, track, and shoot down flying insects in the wild.

  4. 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

  5. Molecular evolution and population genetics of a Gram-negative binding protein gene in the malaria vector Anopheles gambiae (sensu lato)

    OpenAIRE

    Salgueiro, Patrícia; Lopes, Ana Sofia; Mendes, Cristina; Charlwood, Jacques Derek; Arez, Ana Paula; Pinto, João; Silveira, Henrique

    2016-01-01

    Background Clarifying the role of the innate immune system of the malaria vector Anopheles gambiae is a potential way to block the development of the Plasmodium parasites. Pathogen recognition is the first step of innate immune response, where pattern recognition proteins like GNBPs play a central role. Results We analysed 70 sequences of the protein coding gene GNBPB2 from two species, Anopheles gambiae (s.s.) and An. coluzzii, collected in six African countries. We detected 135 segregating ...

  6. 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

    Science.gov (United States)

    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 parasite transmission.

  7. Exploring proteins in Anopheles gambiae male and female antennae through MALDI mass spectrometry profiling.

    Directory of Open Access Journals (Sweden)

    Francesca R Dani

    Full Text Available MALDI profiling and imaging mass spectrometry (IMS are novel techniques for direct analysis of peptides and small proteins in biological tissues. In this work we applied them to the study of Anopheles gambiae antennae, with the aim of analysing expression of soluble proteins involved in olfaction perireceptor events. MALDI spectra obtained by direct profiling on single antennae and by the analysis of extracts, showed similar profiles, although spectra obtained through profiling had a richer ion population and higher signal to noise ratio. Male and female antennae showed distinct protein profiles. MALDI imaging experiments were also performed and differences were observed in the localization of some proteins. Two proteins were identified through high resolution measurement and top-down MS/MS experiments. A 8 kDa protein only present in the male antennae matched with an unannotated sequence of the An. gambiae genome, while the presence of odorant binding protein 9 (OBP-9 was confirmed through experiments of 2-DE, followed by MS and MS/MS analysis of digested spots. This work shows that MALDI MS profiling is a technique suitable for the analysis of proteins of small and medium MW in insect appendices, and allows obtaining data for several specimens which can be investigated for differences between groups. Proteins of interest can be identified through other complementary MS approaches.

  8. 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.

  9. Evidence for X-linked introgression between molecular forms of Anopheles gambiae from Angola.

    Science.gov (United States)

    Choi, K S; Townson, H

    2012-06-01

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

  10. Metabolic and target-site mechanisms combine to confer strong DDT resistance in Anopheles gambiae.

    Directory of Open Access Journals (Sweden)

    Sara N Mitchell

    Full Text Available The development of resistance to insecticides has become a classic exemplar of evolution occurring within human time scales. In this study we demonstrate how resistance to DDT in the major African malaria vector Anopheles gambiae is a result of both target-site resistance mechanisms that have introgressed between incipient species (the M- and S-molecular forms and allelic variants in a DDT-detoxifying enzyme. Sequencing of the detoxification enzyme, Gste2, from DDT resistant and susceptible strains of An. gambiae, revealed a non-synonymous polymorphism (I114T, proximal to the DDT binding domain, which segregated with strain phenotype. Recombinant protein expression and DDT metabolism analysis revealed that the proteins from the susceptible strain lost activity at higher DDT concentrations, characteristic of substrate inhibition. The effect of I114T on GSTE2 protein structure was explored through X-ray crystallography. The amino acid exchange in the DDT-resistant strain introduced a hydroxyl group nearby the hydrophobic DDT-binding region. The exchange does not result in structural alterations but is predicted to facilitate local dynamics and enzyme activity. Expression of both wild-type and 114T alleles the allele in Drosophila conferred an increase in DDT tolerance. The 114T mutation was significantly associated with DDT resistance in wild caught M-form populations and acts in concert with target-site mutations in the voltage gated sodium channel (Vgsc-1575Y and Vgsc-1014F to confer extreme levels of DDT resistance in wild caught An. gambiae.

  11. 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

  12. The knockdown resistance mutation and knockdown time in Anopheles gambiae collected from Mali evaluated through a bottle bioassay and a novel insecticide-treated net bioassay.

    Science.gov (United States)

    Fryxell, Rebecca T Trout; Seifert, Stephanie N; Lee, Yoosook; Sacko, Adama; Lanzaro, Gregory; Cornel, Anthony

    2012-06-01

    Successful malaria management in Mali includes the use of pyrethroids and insecticide-treated nets (ITNs) for mosquito control; however, management is threatened by the spread of insecticide resistance detected via the knockdown resistance (kdr) allele. In a preliminary study, we compared the knockdown times of Anopheles gambiae from Mali using a novel ITN bioassay and the World Health Organization (WHO) bottle bioassay. Additionally, the frequency and relationship between kdr genotypes, molecular forms, and pyrethroid resistance were analyzed. The S molecular form was predominant and accounted for 76% of the assayed population. Both kdr resistant alleles, West Africa resistant (kdr-w) and East Africa resistant (kdr-e), were observed. There was no significant difference in knockdown time based on kdr genotype or molecular form of individual mosquitoes, but mosquitoes in the ITN bioassay homozygous for the kdr-w allele were knocked down significantly faster than those in the WHO bottle bioassay. The ITN bioassay provides an additional indicator of insecticide efficacy because ITNs, frequently used within homes, are the most common form of vector control and malaria prevention, and the ITN bioassays can evaluate seasonal field effects.

  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. Field testing of different chemical combinations as odour baits for trapping wild mosquitoes in The Gambia.

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    Musa Jawara

    Full Text Available Odour baited traps have potential use in population surveillance of insect vectors of disease, and in some cases for vector population reduction. Established attractants for human host-seeking mosquitoes include a combination of CO(2 with L-lactic acid and ammonia, on top of which additional candidate compounds are being tested. In this field study in rural Gambia, using Latin square experiments with thorough randomization and replication, we tested nine different leading candidate combinations of chemical odorants for attractiveness to wild mosquitoes including anthropophilic malaria vectors, using modified Mosquito Magnet-X (MM-X counterflow traps outside experimental huts containing male human sleepers. Highest catches of female mosquitoes, particularly of An. gambiae s.l. and Mansonia species, were obtained by incorporation of tetradecanoic acid. As additional carboxylic acids did not increase the trap catches further, this 'reference blend' (tetradecanoic acid with L-lactic acid, ammonia and CO(2 was used in subsequent experiments. MM-X traps with this blend caught similar numbers of An. gambiae s.l. and slightly more Mansonia and Culex mosquitoes than a standard CDC light trap, and these numbers were not significantly affected by the presence or absence of human sleepers in the huts. Experiments with CO(2 produced from overnight yeast cultures showed that this organic source was effective in enabling trap attractiveness for all mosquito species, although at a slightly lower efficiency than obtained with use of CO(2 gas cylinders. Although further studies are needed to discover additional chemicals that increase attractiveness, as well as to optimise trap design and CO(2 source for broader practical use, the odour-baited traps described here are safe and effective for sampling host-seeking mosquitoes outdoors and can be incorporated into studies of malaria vector ecology.

  15. The Anopheles gambiae detoxification chip: A highly specific microarray to study metabolic-based insecticide resistance in malaria vectors

    OpenAIRE

    2005-01-01

    Metabolic pathways play an important role in insecticide resistance, but the full spectra of the genes involved in resistance has not been established. We constructed a microarray containing unique fragments from 230 Anopheles gambiae genes putatively involved in insecticide metabolism [cytochrome P450s (P450s), GSTs, and carboxylesterases and redox genes, partners of the P450 oxidative metabolic complex, and various controls]. We used this detox chip to monitor the expression of the detoxify...

  16. Spatio-temporal variations of Anopheles coluzzii and An. gambiae and their Plasmodium infectivity rates in Lobito, Angola.

    Science.gov (United States)

    Carnevale, Pierre; Toto, Jean-Claude; Besnard, Patrick; Santos, Maria Adelaide Dos; Fortes, Filomeno; Allan, Richard; Manguin, Sylvie

    2015-06-01

    From 2003 to 2007, entomological surveys were conducted in Lobito town (Benguela Province, Angola) to determine which Anopheles species were present and to identify the vectors responsible for malaria transmission in areas where workers of the Sonamet Company live. Two types of surveys were conducted: (1) time and space surveys in the low and upper parts of Lobito during the rainy and dry periods; (2) a two-year longitudinal study in Sonamet workers' houses provided with long-lasting insecticide-treated nets (LLIN), "PermaNet," along with the neighboring community. Both species, An. coluzzii (M molecular form) and An. gambiae (S molecular form), were collected. Anopheles coluzzii was predominant during the dry season in the low part of Lobito where larvae develop in natural ponds and temporary pools. However, during the rainy season, An. gambiae was found in higher proportions in the upper part of the town where larvae were collected in domestic water tanks built near houses. Anopheles melas and An. listeri were captured in higher numbers during the dry season and in the low part of Lobito where larvae develop in stagnant brackish water pools. The infectivity rates of An. gambiae s.l. varied from 0.90% to 3.41%.

  17. Evidence that agricultural use of pesticides selects pyrethroid resistance within Anopheles gambiae s.l. populations from cotton growing areas in Burkina Faso, West Africa

    Science.gov (United States)

    Hien, Aristide Sawdetuo; Soma, Dieudonné Diloma; Hema, Omer; Bayili, Bazoma; Namountougou, Moussa; Gnankiné, Olivier; Baldet, Thierry; Diabaté, Abdoulaye; Dabiré, Kounbobr Roch

    2017-01-01

    Many studies have shown the role of agriculture in the selection and spread of resistance of Anopheles gambiae s.l. to insecticides. However, no study has directly demonstrated the presence of insecticides in breeding sources as a source of selection for this resistance. It is in this context that we investigated the presence of pesticide residues in breeding habitats and their formal involvement in vector resistance to insecticides in areas of West Africa with intensive farming. This study was carried out from June to November 2013 in Dano, southwest Burkina Faso in areas of conventional (CC) and biological cotton (BC) growing. Water and sediment samples collected from breeding sites located near BC and CC fields were submitted for chromatographic analysis to research and titrate the residual insecticide content found there. Larvae were also collected in these breeding sites and used in toxicity tests to compare their mortality to those of the susceptible strain, Anopheles gambiae Kisumu. All tested mosquitoes (living and dead) were analyzed by PCR for species identification and characterization of resistance genes. The toxicity analysis of water from breeding sites showed significantly lower mortality rates in breeding site water from biological cotton (WBC) growing sites compared to that from conventional cotton (WCC) sites respective to both An. gambiae Kisumu (WBC: 80.75% vs WCC: 92.75%) and a wild-type strain (49.75% vs 66.5%). The allele frequencies L1014F, L1014S kdr, and G116S ace -1R mutations conferring resistance, respectively, to pyrethroids and carbamates / organophosphates were 0.95, 0.4 and 0.12. Deltamethrin and lambda-cyhalothrin were identified in the water samples taken in October/November from mosquitoes breeding in the CC growing area. The concentrations obtained were respectively 0.0147ug/L and 1.49 ug/L to deltamethrin and lambdacyhalothrin. Our results provided evidence by direct analysis (biological and chromatographic tests) of the role

  18. Identification and analysis of Single Nucleotide Polymorphisms (SNPs in the mosquito Anopheles funestus, malaria vector

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    Hemingway Janet

    2007-01-01

    Full Text Available Abstract Background Single nucleotide polymorphisms (SNPs are the most common source of genetic variation in eukaryotic species and have become an important marker for genetic studies. The mosquito Anopheles funestus is one of the major malaria vectors in Africa and yet, prior to this study, no SNPs have been described for this species. Here we report a genome-wide set of SNP markers for use in genetic studies on this important human disease vector. Results DNA fragments from 50 genes were amplified and sequenced from 21 specimens of An. funestus. A third of specimens were field collected in Malawi, a third from a colony of Mozambican origin and a third form a colony of Angolan origin. A total of 494 SNPs including 303 within the coding regions of genes and 5 indels were identified. The physical positions of these SNPs in the genome are known. There were on average 7 SNPs per kilobase similar to that observed in An. gambiae and Drosophila melanogaster. Transitions outnumbered transversions, at a ratio of 2:1. The increased frequency of transition substitutions in coding regions is likely due to the structure of the genetic code and selective constraints. Synonymous sites within coding regions showed a higher polymorphism rate than non-coding introns or 3' and 5'flanking DNA with most of the substitutions in coding regions being observed at the 3rd codon position. A positive correlation in the level of polymorphism was observed between coding and non-coding regions within a gene. By genotyping a subset of 30 SNPs, we confirmed the validity of the SNPs identified during this study. Conclusion This set of SNP markers represents a useful tool for genetic studies in An. funestus, and will be useful in identifying candidate genes that affect diverse ranges of phenotypes that impact on vector control, such as resistance insecticide, mosquito behavior and vector competence.

  19. 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

  20. Kinetic properties of alternatively spliced isoforms of laccase-2 from Tribolium castaneum and Anopheles gambiae.

    Science.gov (United States)

    Gorman, Maureen J; Sullivan, Lucinda I; Nguyen, Thi D T; Dai, Huaien; Arakane, Yasuyuki; Dittmer, Neal T; Syed, Lateef U; Li, Jun; Hua, Duy H; Kanost, Michael R

    2012-03-01

    Laccase-2 is a highly conserved multicopper oxidase that functions in insect cuticle pigmentation and tanning. In many species, alternative splicing gives rise to two laccase-2 isoforms. A comparison of laccase-2 sequences from three orders of insects revealed eleven positions at which there are conserved differences between the A and B isoforms. Homology modeling suggested that these eleven residues are not part of the substrate binding pocket. To determine whether the isoforms have different kinetic properties, we compared the activity of laccase-2 isoforms from Tribolium castaneum and Anopheles gambiae. We partially purified the four laccases as recombinant enzymes and analyzed their ability to oxidize a range of laccase substrates. The predicted endogenous substrates tested were dopamine, N-acetyldopamine (NADA), N-β-alanyldopamine (NBAD) and dopa, which were detected in T. castaneum previously and in A. gambiae as part of this study. Two additional diphenols (catechol and hydroquinone) and one non-phenolic substrate (2,2'-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid)) were also tested. We observed no major differences in substrate specificity between the A and B isoforms. Dopamine, NADA and NBAD were oxidized with catalytic efficiencies ranging from 51 to 550 min⁻¹ mM⁻¹. These results support the hypothesis that dopamine, NADA and NBAD are endogenous substrates for both isoforms of laccase-2. Catalytic efficiencies associated with dopa oxidation were low, ranging from 8 to 30 min⁻¹ mM⁻¹; in comparison, insect tyrosinase oxidized dopa with a catalytic efficiency of 201 min⁻¹ mM⁻¹. We found that dopa had the highest redox potential of the four endogenous substrates, and this property of dopa may explain its poor oxidation by laccase-2. We conclude that laccase-2 splice isoforms are likely to oxidize the same substrates in vivo, and additional experiments will be required to discover any isoform-specific functions.

  1. Attractiveness of MM-X traps baited with human or synthetic odor to mosquitoes (Diptera: Culicidae) in The Gambia

    NARCIS (Netherlands)

    Qiu, Y.T.; Smallegange, R.C.; Braak, ter C.J.F.; Spitzen, J.; Loon, van J.J.A.; Jawara, M.; Milligan, P.; Galimard, A.M.S.; Beek, van T.A.; Knols, B.G.J.; Takken, W.

    2007-01-01

    Chemical cues play an important role in the host-seeking behavior of blood-feeding mosquitoes (Diptera: Culicidae). A field study was carried out in The Gambia to investigate the effects of human odor or synthetic odor blends on the attraction of mosquitoes. MM-X traps baited with 16 odor blends to

  2. 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

  3. 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.

  4. Using a near-infrared spectrometer to estimate the age of Anopheles mosquitoes exposed to pyrethroids

    Science.gov (United States)

    We report on the accuracy of using near-infrared spectroscopy (NIRS) to predict the age of Anopheles mosquitoes reared from wild larvae and a mixed age-wild adult population collected from pit traps after exposure to pyrethroids. The mosquitoes reared from wild larvae were estimated as ,7 or $7 d ol...

  5. 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.

  6. Microbial larvicides for malaria control in The Gambia.

    OpenAIRE

    Green Clare; Lindsay Steven W; Majambere Silas; Kandeh Balla; Fillinger Ulrike

    2007-01-01

    Abstract Background Mosquito larval control may prove to be an effective tool for incorporating into integrated vector management (IVM) strategies for reducing malaria transmission. Here the efficacy of microbial larvicides against Anopheles gambiae s.l. was tested in preparation for a large-scale larviciding programme in The Gambia. Methods The impact of water-dispersible (WDG) and corn granule (CG) formulations of commercial Bacillus sphaericus strain 2362 (Bs; VectoLex®) and Bacillus thuri...

  7. Distribution of the species of the Anopheles gambiae complex and first evidence of Anopheles merus as a malaria vector in Madagascar

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    Le Goff Gilbert

    2003-10-01

    Full Text Available Abstract Background Members of the Anopheles gambiae complex are amongst the best malaria vectors in the world, but their vectorial capacities vary between species and populations. A large-scale sampling of An. gambiae sensu lato was carried out in various bioclimatic domains of Madagascar. Local abundance of an unexpected member of this complex raised questions regarding its role in malaria transmission. Methods Sampling took place at 38 sites and 2,067 females were collected. Species assessment was performed using a PCR targeting a sequence in the IGS of the rDNA. Analysis focused on the relative prevalence of the species per site, bioclimatic domain and altitude. Infectivity of Anopheles merus was assessed using an ELISA to detect the presence of malarial circumsporozoite protein in the head-thorax. Results Three species were identified: An. gambiae, Anopheles arabiensis and An. merus. The distribution of each species is mainly a function of bioclimatic domains and, to a lesser extent, altitude. An. arabiensis is present in all bioclimatic domains with highest prevalence in sub-humid, dry and sub-arid domains. An. gambiae has its highest prevalence in the humid domain, is in the minority in dry areas, rare in sub-humid and absent in sub-arid domains. An. merus is restricted to the coastal fringe in the south and west; it was in the majority in one southern village. The majority of sites were sympatric for at least two of the species (21/38 and two sites harboured all three species. The role of An. merus as malaria vector was confirmed in the case of two human-biting females, which were ELISA-positive for Plasmodium falciparum. Conclusion Despite the huge environmental (mainly man-made changes in Madagascar, the distribution of An. gambiae and An. arabiensis appears unchanged for the past 35 years. The distribution of An. merus is wider than was previously known, and its effectiveness as a malaria vector has been shown for the first time; this

  8. Malaria vectors in the Republic of Benin: distribution of species and molecular forms of the Anopheles gambiae complex.

    Science.gov (United States)

    Djogbénou, Luc; Pasteur, Nicole; Bio-Bangana, Sahabi; Baldet, Thierry; Irish, Seth R; Akogbeto, Martin; Weill, Mylène; Chandre, Fabrice

    2010-05-01

    Members of the Anopheles gambiae complex are among the best malaria vectors in the world, but their vectorial capacities vary between species and populations. A large-scale sampling of An. gambiae sensu lato was carried out in 2006 and 2007 in various bioclimatic areas of Benin (West Africa). The objective of this study was to collate data on the relative frequencies of species and forms within the An. gambiae complex and to produce a map of their spatial distribution. Sampling took place at 30 sites and 2122 females were analyzed. Two species were identified through molecular methods. The overall collection showed a preponderance of An. gambiae s.s., but unexpectedly, An. arabiensis was reported in the coastal-Guinean bioclimatic area characterized by a mean annual rainfall of >1500 mm where only An. gambiae s.s. was reported previously. Our study of Benin indicates that An. arabiensis would be adapted not only to the urban areas but also to the rural humid regions. Among 1717 An. gambiae s.s., 26.5% were of the M form and 73.3% were S form. Few hybrid specimens between the M and S forms were observed (0.2%). Only the spatial distribution of the M form appears to be mainly a function of bioclimatic area. Factors that influence the distribution of these malaria vectors are discussed. This study underlines the need of further investigations of biological, ecological, and behavioral traits of these species and forms to better appreciate their vectorial capacities. Acquisition of entomological field data appears essential to better estimate the stratification of malaria risk and help improve malaria vector control interventions.

  9. Bioefficacy of long-lasting insecticidal nets against pyrethroid-resistant populations of Anopheles gambiae s.s. from different malaria transmission zones in Uganda

    OpenAIRE

    Okia, Michael; Ndyomugyenyi, Richard; Kirunda, James; Byaruhanga, Anatol; Adibaku, Seraphine; Lwamafa, Denis K; Kironde, Fred

    2013-01-01

    Background There are major concerns over sustaining the efficacy of current malaria vector control interventions given the rapid spread of resistance, particularly to pyrethroids. This study assessed the bioefficacy of five WHO-recommended long-lasting insecticidal nets (LLINs) against pyrethroid-resistant Anopheles gambiae field populations from Uganda. Methods Adult An. gambiae from Lira, Tororo, Wakiso and Kanungu districts were exposed to permethrin (0.75%) or deltamethrin (0.05%) in stan...

  10. The effect of water physical quality and water level changes on the occurrence and density of larvae of Anopheles mosquitoes around the shoreline of the Koka reservoir, Central Ethiopia

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    S. Kibret

    2010-08-01

    Full Text Available Entomological studies to determine the effect of the physical characteristics of larval breeding water bodies and reservoir water level changes on the occurrence of Anopheles mosquito larvae and on the spatial and temporal formation of larval breeding habitats were conducted in two villages at Koka reservoir between August and December 2007. Of the two study villages, Ejersa is in close proximity to the reservoir, and Kuma is 5 km away from it. Data on the type, number and physical characteristics of Anopheles larval breeding habitat, species composition and densities of anopheles mosquitoes in and around the study villages were investigated and recorded. Meteorological and reservoir water level data were compared with availability of Anopheles larval breeding sites and densities. Entomological data from the weekly larval collections showed that Anopheles pharoensis Theobald, Anopheles gambiae s.l. Giles, Anopheles coustani Laveran and Anopheles squamosus Theobald were breeding in the study area. The mean larval density of A. gambiae s.l. in this study was higher in slightly turbid and shallow aquatic habitats than in turbid and relatively deep aquatic habitats (F=16.97, pF=6.03, pA. pharoensis in habitat with floating vegetation and with relatively shady conditions was significantly higher than that of less shaded aquatic habitat and greater emergent vegetation (F=15.75, pF=10.56, pAnopheles larvae with water temperature of the breeding habitat and daily minimum atmospheric temperature (r=0.541, pr=0.604, pr=0.605, pAnopheles larval count, and the density of Anopheles mosquitoes in the vicinity. The proliferation of suitable breeding habitats around the reservoir villages is strongly associated with reservoir water level changes. This is particularly important for A. pharoensis and A. arabiensis which are important vectors of malaria in the area. Further investigation on the species diversity, physical and chemical habitat characteristics and

  11. Mosquito biosurveillance on Kyushu Island, Japan, with emphasis on Anopheles Hyrcanus Group and related species (Diptera: culicidae).

    Science.gov (United States)

    Rueda, Leopoldo M; Pagac, Benedict; Iwakami, Masashiro; Spring, Alexandra R; Motoki, Mayasa T; Pecor, James E; Higa, Yukiko; Futami, Kyoko; Imanishi, Nozomi; Long, Lewis S; Debboun, Mustapha

    2014-01-01

    This report includes the distribution records of the Anopheles (Anopheles) Hyrcanus Group and associated species in Kyushu Island, Japan, based on our field collections from various localities of 4 prefectures (Fukuoka, Kumamoto, Nagasaki, Saga), primarily from 2002-2013. The status of common and potential mosquito vectors, particularly Anopheles species, in Japan are noted.

  12. Diversity of the Bacterial Microbiota of Anopheles Mosquitoes from Binh Phuoc Province, Vietnam.

    Science.gov (United States)

    Ngo, Chung T; Romano-Bertrand, Sara; Manguin, Sylvie; Jumas-Bilak, Estelle

    2016-01-01

    The naturally acquired microbiota of Anopheles can influence vector's susceptibility to Plasmodium and its capacity to transmit them. Microbiota modification is a new challenge to limit disease transmission but it still needs advanced knowledges on bacterial community in Anopheles, especially in wild and infected specimens from diverse origin and species. Bacterial culture and 16S rRNA gene-PCR associated to Temporal Temperature Gradient Electrophoresis (TTGE) were applied to explore the bacterial diversity in the abdomen of 100 wild specimens (eight Anopheles species) collected in the Binh Phuoc Province, Vietnam. Culture and PCR-TTGE were complementary. The bacterial richness of the mosquito collection encompassed 105 genera belonging to seven phyla, mostly Proteobacteria, Firmicutes, and Actinobacteria. Staphylococcus, Clostridium, and Bacillus in Firmicutes were the most prevalent genera. However, Proteobacteria represented by 57 genera was the most diversified phylum in Anopheles microbiota. The high overall of Anopheles-associated bacteria is confirmed with, to our knowledge, 51 genera described for the first time in Anopheles microbiota. However, the diversity per specimen was low with average diversity index and the average Shannon-Wiener score (H) of 4.843 and 5.569, respectively. The most represented bacterial genera were present in microbiota share by Anopheles from Binh Phuoc was very narrow, suggesting that Anopheles microbiota was greatly influenced by local environments. The repertory of bacterial genera in two specimens of An. dirus and An. pampanai naturally infected by Plasmodium vivax was also described as preliminary results. Finally, this study completed the repertory of bacteria associated to wild Anopheles. Anopheles associated-bacteria appeared specimen-dependent rather than mosquitoe species- or group-dependent. Their origin and the existence of Anopheles-specific bacterial taxa are discussed.

  13. Wash resistance and repellent properties of Africa University mosquito blankets against mosquitoes

    OpenAIRE

    N. Lukwa; A. Makuwaza; T. Chiwade; Mutambu, S L; M. Zimba; P. Munosiyei

    2013-01-01

    The effect of permethrin-treated Africa University (AU) mosquito blankets on susceptible female Anopheles gambiae sensu lato mosquitoes was studied under laboratory conditions at Africa University Campus in Mutare, Zimbabwe. Wash resistance (ability to retain an effective dose that kills ≥80% of mosquitoes after a number of washes) and repellence (ability to prevent ≥80% of mosquito bites) properties were studied. The AU blankets were wash resistant when 100% mortality was recorded up t...

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

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    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

  15. Mitochondrial NAD+-dependent malic enzyme from Anopheles stephensi: a possible novel target for malaria mosquito control

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    Pon Jennifer

    2011-10-01

    Full Text Available Abstract Background Anopheles stephensi mitochondrial malic enzyme (ME emerged as having a relevant role in the provision of pyruvate for the Krebs' cycle because inhibition of this enzyme results in the complete abrogation of oxygen uptake by mitochondria. Therefore, the identification of ME in mitochondria from immortalized A. stephensi (ASE cells and the investigation of the stereoselectivity of malate analogues are relevant in understanding the physiological role of ME in cells of this important malaria parasite vector and its potential as a possible novel target for insecticide development. Methods To characterize the mitochondrial ME from immortalized ASE cells (Mos. 43; ASE, mass spectrometry analyses of trypsin fragments of ME, genomic sequence analysis and biochemical assays were performed to identify the enzyme and evaluate its activity in terms of cofactor dependency and inhibitor preference. Results The encoding gene sequence and primary sequences of several peptides from mitochondrial ME were found to be highly homologous to the mitochondrial ME from Anopheles gambiae (98% and 59% homologous to the mitochondrial NADP+-dependent ME isoform from Homo sapiens. Measurements of ME activity in mosquito mitochondria isolated from ASE cells showed that (i Vmax with NAD+ was 3-fold higher than that with NADP+, (ii addition of Mg2+ or Mn2+ increased the Vmax by 9- to 21-fold, with Mn2+ 2.3-fold more effective than Mg2+, (iii succinate and fumarate increased the activity by 2- and 5-fold, respectively, at sub-saturating concentrations of malate, (iv among the analogs of L-malate tested as inhibitors of the NAD+-dependent ME catalyzed reaction, small (2- to 3-carbons organic diacids carrying a 2-hydroxyl/keto group behaved as the most potent inhibitors of ME activity (e.g., oxaloacetate, tartronic acid and oxalate. Conclusions The biochemical characterization of Anopheles stephensi ME is of critical relevance given its important role in

  16. 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

  17. Behavioural responses of Anopheles gambiae sensu stricto to components of human breath, sweat and urine depend on mixture composition and concentration

    NARCIS (Netherlands)

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

    2011-01-01

    Host-seeking behaviour of the anthropophilic malaria vector Anopheles gambiae sensu stricto (Diptera: Culicidae) is mediated predominantly by olfactory cues. Several hundreds of odour components have been identified from human emanations, but only a few have been proven to act as attractants or syne

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

    Science.gov (United States)

    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.

  19. 1.45 Å resolution structure of SRPN18 from the malaria vector Anopheles gambiae

    Energy Technology Data Exchange (ETDEWEB)

    Meekins, David A.; Zhang, Xin; Battaile, Kevin P.; Lovell, Scott; Michel, Kristin (Kansas); (KSU); (HWMRI)

    2016-11-19

    Serine protease inhibitors (serpins) in insects function within development, wound healing and immunity. The genome of the African malaria vector,Anopheles gambiae, encodes 23 distinct serpin proteins, several of which are implicated in disease-relevant physiological responses.A. gambiaeserpin 18 (SRPN18) was previously categorized as non-inhibitory based on the sequence of its reactive-center loop (RCL), a region responsible for targeting and initiating protease inhibition. The crystal structure ofA. gambiaeSRPN18 was determined to a resolution of 1.45 Å, including nearly the entire RCL in one of the two molecules in the asymmetric unit. The structure reveals that the SRPN18 RCL is extremely short and constricted, a feature associated with noncanonical inhibitors or non-inhibitory serpin superfamily members. Furthermore, the SRPN18 RCL does not contain a suitable protease target site and contains a large number of prolines. The SRPN18 structure therefore reveals a unique RCL architecture among the highly conserved serpin fold.

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

    Directory of Open Access Journals (Sweden)

    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.

  1. 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

  2. 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.

  3. Discovery of flavivirus-derived endogenous viral elements in Anopheles mosquito genomes supports the existence of Anopheles-associated insect-specific flaviviruses.

    Science.gov (United States)

    Lequime, Sebastian; Lambrechts, Louis

    2017-01-01

    The Flavivirus genus encompasses several arboviruses of public health significance such as dengue, yellow fever, and Zika viruses. It also includes insect-specific flaviviruses (ISFs) that are only capable of infecting insect hosts. The vast majority of mosquito-infecting flaviviruses have been associated with mosquito species of the Aedes and Culex genera in the Culicinae subfamily, which also includes most arbovirus vectors. Mosquitoes of the Anophelinae subfamily are not considered significant arbovirus vectors; however, flaviviruses have occasionally been detected in field-caught Anopheles specimens. Whether such observations reflect occasional spillover or laboratory contamination or whether Anopheles mosquitoes are natural hosts of flaviviruses is unknown. Here, we provide in silico and in vivo evidence of transcriptionally active, flavivirus-derived endogenous viral elements (EVEs) in the genome of Anopheles minimus and Anopheles sinensis. Such non-retroviral endogenization of RNA viruses is consistent with a shared evolutionary history between flaviviruses and Anopheles mosquitoes. Phylogenetic analyses of the two newly described EVEs support the existence of a distinct clade of Anopheles-associated ISFs.

  4. 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

  5. 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

  6. 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

  7. Cultured skin microbiota attracts malaria mosquitoes

    OpenAIRE

    Takken Willem; Knols Bart GJ; Beijleveld Hans; Verhulst Niels O; Schraa Gosse; Bouwmeester Harro J; Smallegange Renate C

    2009-01-01

    Abstract Background Host-seeking of the African malaria mosquito, Anopheles gambiae sensu stricto, is guided by human odours. The precise nature of the odours, and the composition of attractive blends of volatiles, remains largely unknown. Skin microbiota plays an important role in the production of human body odours. It is hypothesized that host attractiveness and selection of An. gambiae is affected by the species composition, density, and metabolic activity of the skin microbiota. A study ...

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

    Directory of Open Access Journals (Sweden)

    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.

  9. Fitness of Transgenic Anopheles stephensi Mosquitoes Expressing the SM1 Peptide under the Control of a Vitellogenin Promoter

    OpenAIRE

    Li, Chaoyang; Marrelli, Mauro T.; Yan, Guiyun; Jacobs-Lorena, Marcelo

    2008-01-01

    Three transgenic Anopheles stephensi lines were established that strongly inhibit transmission of the mouse malaria parasite Plasmodium berghei. Fitness of the transgenic mosquitoes was assessed based on life table analysis and competition experiments between transgenic and wild-type mosquitoes. Life table analysis indicated low fitness load for the 2 single-insertion transgenic mosquito lines VD35 and VD26 and no load for the double-insertion transgenic mosquito line VD9. However, in cage ex...

  10. Sperm quantity and size variation in un-irradiated and irradiated males of the malaria mosquito Anopheles arabiensis Patton

    NARCIS (Netherlands)

    Helinski, M.; Knols, B.G.J.

    2009-01-01

    Anopheles mosquitoes are important candidates for genetic control strategies. However, little is known about sperm quality and quantity as determinants of male reproductive success. In this study, sperm quantity and length variation were assessed in testes of un-irradiated and irradiated Anopheles a

  11. Exogenous gypsy insulator sequences modulate transgene expression in the malaria vector mosquito, Anopheles stephensi.

    Science.gov (United States)

    Carballar-Lejarazú, Rebeca; Jasinskiene, Nijole; James, Anthony A

    2013-04-30

    Malaria parasites are transmitted to humans by mosquitoes of the genus Anopheles, and these insects are the targets of innovative vector control programs. Proposed approaches include the use of genetic strategies based on transgenic mosquitoes to suppress or modify vector populations. Although substantial advances have been made in engineering resistant mosquito strains, limited efforts have been made in refining mosquito transgene expression, in particular attenuating the effects of insertions sites, which can result in variations in phenotypes and impacts on fitness due to the random integration of transposon constructs. A promising strategy to mitigate position effects is the identification of insulator or boundary DNA elements that could be used to isolate transgenes from the effects of their genomic environment. We applied quantitative approaches that show that exogenous insulator-like DNA derived from the Drosophila melanogaster gypsy retrotransposon can increase and stabilize transgene expression in transposon-mediated random insertions and recombinase-catalyzed, site-specific integrations in the malaria vector mosquito, Anopheles stephensi. These sequences can contribute to precise expression of transgenes in mosquitoes engineered for both basic and applied goals.

  12. Cloning and characterization of two glutathione S-transferases from a DDT-resistant strain of Anopheles gambiae.

    Science.gov (United States)

    Ranson, H; Prapanthadara, L a; Hemingway, J

    1997-05-15

    Two cDNA species, aggst1-5 and aggst1-6, comprising the entire coding region of two distinct glutathione S-transferases (GSTs) have been isolated from a 1,1,1-trichloro-2,2-bis-(p-chlorophenyl)ethane (DDT) resistant strain (ZANDS) of Anopheles gambiae. The nucleotide sequences of these cDNA species share 80.2% identity and their derived amino acid sequences are 82.3% similar. They have been classified as insect class I GSTs on the basis of their high sequence similarity to class I GSTs from Drosophila melanogaster and Musca domestica and they are localized to a region of an An. gambiae chromosome known to contain further class I GSTs. The genes aggst1-5 and aggst1-6 were expressed at high levels in Escherichia coli and the recombinant GSTs were purified by affinity chromatography and characterized. Both agGST1-5 and agGST1-6 showed high activity with the substrates 1-chloro-2,4-dinitrobenzene and 1, 2-dichloro-4-nitrobenzene but negligible activity with the mammalian theta class substrates, 1,2-epoxy-3-(4-nitrophenoxy)propane and p-nitrophenyl bromide. Despite their high level of sequence identity, agGST1-5 and agGST1-6 displayed different kinetic properties. Both enzymes were able to metabolize DDT and were localized to a subset of GSTs that, from earlier biochemical studies, are known to be involved in insecticide resistance in An. gambiae. This subset of enzymes is one of three in which the DDT metabolism levels are elevated in resistant insects.

  13. Cultured skin microbiota attracts malaria mosquitoes

    NARCIS (Netherlands)

    Verhulst, N.O.; Beijleveld, H.; Knols, B.G.J.; Takken, W.; Schraa, G.; Bouwmeester, H.J.; Smallegange, R.C.

    2009-01-01

    Background - Host-seeking of the African malaria mosquito, Anopheles gambiae sensu stricto, is guided by human odours. The precise nature of the odours, and the composition of attractive blends of volatiles, remains largely unknown. Skin microbiota plays an important role in the production of human

  14. 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.

  15. Wash resistance and repellent properties of Africa University mosquito blankets against mosquitoes

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    N. Lukwa

    2013-04-01

    Full Text Available The effect of permethrin-treated Africa University (AU mosquito blankets on susceptible female Anopheles gambiae sensu lato mosquitoes was studied under laboratory conditions at Africa University Campus in Mutare, Zimbabwe. Wash resistance (ability to retain an effective dose that kills ≥80% of mosquitoes after a number of washes and repellence (ability to prevent ≥80% of mosquito bites properties were studied. The AU blankets were wash resistant when 100% mortality was recorded up to 20 washes, declining to 90% after 25 washes. Untreated AU blankets did not cause any mortality on mosquitoes. However, mosquito repellence was 96%, 94%, 97.9%, 87%, 85% and 80.7% for treated AU blankets washed 0, 5, 10, 15, 20 and 25 times, respectively. Mosquito repellence was consistently above 80% from 0-25 washes. In conclusion, AU blankets washed 25 times were effective in repelling and killing An. gambiae sl mosquitoes under laboratory conditions.

  16. Mosquito transgenic technologies to reduce Plasmodium transmission.

    Science.gov (United States)

    Fuchs, Silke; Nolan, Tony; Crisanti, Andrea

    2013-01-01

    The ability to introduce genetic constructs of choice into the genome of Anopheles mosquitoes provides a valuable tool to study the molecular interactions between the Plasmodium parasite and its insect host. In the long term, this technology could potentially offer new ways to control vector-borne diseases through the suppression of target mosquito populations or through the introgression of traits that preclude pathogen transmission. Here, we describe in detail protocols for the generation of transgenic Anopheles gambiae mosquitoes based on germ-line transformation using either modified transposable elements or the site-specific PhiC31 recombinase.

  17. 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.

  18. Susceptibility Status of Anopheles sundaicus Mosquitoes Against Insecticides Cypermethrin in Garut Regency

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    Nunung Seniawati

    2010-06-01

    Full Text Available At the time of high vector populations and malaria transmission is in progress, it is necessary to use insecticides to control vector using the house spraying. To get the results as objective the eradication of the vector that is able to suppress the vector population so that no longer play a role in malaria transmission, the insecticide used should be effective against mosquitoes and the mosquitoes are still susceptible to the insecticide used. To determine the level of malaria vector mosquito susceptibility to insecticides, in the terri-tory of Garut district has conducted susceptibility tests of Anopheles sundaicus mosquitoes to insecticides Cypermethrin held in November up to December 2008. Mosquitoes tested were captured in the form of larvae from ponds and estuaries in Karyamukti Cibalong Garut, and then reared in the field insektarium. The adult level were then tested for their susceptibility. Mosquito susceptibility tests conducted using the WHO Susceptibility Test Kit as many as four repetitions performed simultaneously, while the insecticide used in the form of imprag-nated paper with a dose of 0.05%. From tests it is known that mosquito mortality rate up to 100% test. This indicates that the mosquito An. sundaicus in Garut regency of West Java, is still susceptible to the insecticide Cypermenthrin. Therefore, it can still be used in the eradication of malaria vectors in the recommended dosage of 0.20 g/m2.

  19. Bacterial diversity associated with wild caught Anopheles mosquitoes from Dak Nong Province, Vietnam using culture and DNA fingerprint.

    Directory of Open Access Journals (Sweden)

    Chung Thuy Ngo

    Full Text Available Microbiota of Anopheles midgut can modulate vector immunity and block Plasmodium development. Investigation on the bacterial biodiversity in Anopheles, and specifically on the identification of bacteria that might be used in malaria transmission blocking approaches, has been mainly conducted on malaria vectors of Africa. Vietnam is an endemic country for both malaria and Bancroftian filariasis whose parasitic agents can be transmitted by the same Anopheles species. No information on the microbiota of Anopheles mosquitoes in Vietnam was available previous to this study.The culture dependent approach, using different mediums, and culture independent (16S rRNA PCR - TTGE method were used to investigate the bacterial biodiversity in the abdomen of 5 Anopheles species collected from Dak Nong Province, central-south Vietnam. Molecular methods, sequencing and phylogenetic analysis were used to characterize the microbiota.The microbiota in wild-caught Anopheles was diverse with the presence of 47 bacterial OTUs belonging to 30 genera, including bacterial genera impacting Plasmodium development. The bacteria were affiliated with 4 phyla, Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria, the latter being the dominant phylum. Four bacterial genera are newly described in Anopheles mosquitoes including Coxiella, Yersinia, Xanthomonas, and Knoellia. The bacterial diversity per specimen was low ranging from 1 to 4. The results show the importance of pairing culture and fingerprint methods to better screen the bacterial community in Anopheles mosquitoes.Sampled Anopheles species from central-south Vietnam contained a diverse bacterial microbiota that needs to be investigated further in order to develop new malaria control approaches. The combination of both culture and DNA fingerprint methods allowed a thorough and complementary screening of the bacterial community in Anopheles mosquitoes.

  20. Bacterial Diversity Associated with Wild Caught Anopheles Mosquitoes from Dak Nong Province, Vietnam Using Culture and DNA Fingerprint

    Science.gov (United States)

    Ngo, Chung Thuy; Aujoulat, Fabien; Veas, Francisco; Jumas-Bilak, Estelle; Manguin, Sylvie

    2015-01-01

    Background Microbiota of Anopheles midgut can modulate vector immunity and block Plasmodium development. Investigation on the bacterial biodiversity in Anopheles, and specifically on the identification of bacteria that might be used in malaria transmission blocking approaches, has been mainly conducted on malaria vectors of Africa. Vietnam is an endemic country for both malaria and Bancroftian filariasis whose parasitic agents can be transmitted by the same Anopheles species. No information on the microbiota of Anopheles mosquitoes in Vietnam was available previous to this study. Method The culture dependent approach, using different mediums, and culture independent (16S rRNA PCR – TTGE) method were used to investigate the bacterial biodiversity in the abdomen of 5 Anopheles species collected from Dak Nong Province, central-south Vietnam. Molecular methods, sequencing and phylogenetic analysis were used to characterize the microbiota. Results and Discussion The microbiota in wild-caught Anopheles was diverse with the presence of 47 bacterial OTUs belonging to 30 genera, including bacterial genera impacting Plasmodium development. The bacteria were affiliated with 4 phyla, Actinobacteria, Bacteroidetes, Firmicutes and Proteobacteria, the latter being the dominant phylum. Four bacterial genera are newly described in Anopheles mosquitoes including Coxiella, Yersinia, Xanthomonas, and Knoellia. The bacterial diversity per specimen was low ranging from 1 to 4. The results show the importance of pairing culture and fingerprint methods to better screen the bacterial community in Anopheles mosquitoes. Conclusion Sampled Anopheles species from central-south Vietnam contained a diverse bacterial microbiota that needs to be investigated further in order to develop new malaria control approaches. The combination of both culture and DNA fingerprint methods allowed a thorough and complementary screening of the bacterial community in Anopheles mosquitoes. PMID:25747513

  1. Mosquito Larvicidal Potential of Gossypium hirsutum (Bt cotton) Leaves Extracts against Aedes aegypti and Anopheles stephensi larvae.

    OpenAIRE

    Patil, Chandrashekhar D; Hemant P Borase; Salunkhe, Rahul B; Rahul K Suryawanshi; Narkhade, Chandrakant P; Salunke, Bipinchandra K.; Satish V Patil

    2014-01-01

    Background: We aimed to extract the ingredients from leaves of Gossypium hirsutum (Bt cotton) using different solvents and evaluate for potential use to control different larval stages of mosquito species, Aedes aegypti and Anopheles stephensi. Methods: Qualitative and quantitative estimation of ingredients from Go. hirsutum (Bt) plant extract was carried out and their inhibitory action against mosquito larvae was determined using mosquito larvicidal assay. Results: LC50 values of water, etha...

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

    Directory of Open Access Journals (Sweden)

    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

  3. 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.

  4. 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.

  5. A low-cost microfluidic chip for rapid genotyping of malaria-transmitting mosquitoes.

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    Changchun Liu

    Full Text Available BACKGROUND: Vector control is one of the most effective measures to prevent the transmission of malaria, a disease that causes over 600,000 deaths annually. Around 30-40 Anopheles mosquito species are natural vectors of malaria parasites. Some of these species cannot be morphologically distinguished, but have behavioral and ecological differences. Emblematic of this is the Anopheles gambiae species complex. The correct identification of vector species is fundamental to the development of control strategies and epidemiological studies of disease transmission. METHODOLOGY/PRINCIPAL FINDINGS: An inexpensive, disposable, field-deployable, sample-to-answer, microfluidic chip was designed, constructed, and tested for rapid molecular identification of Anopheles gambiae and Anopheles arabiensis. The chip contains three isothermal amplification reactors. One test reactor operates with specific primers to amplify Anopheles gambiae DNA, another with specific primers for Anopheles arabiensis DNA, and the third serves as a negative control. A mosquito leg was crushed on an isolation membrane. Two discs, laden with mosquito tissue, were punched out of the membrane and inserted into the two test chambers. The isolated, disc-bound DNA served as a template in the amplification processes. The amplification products were detected with intercalating fluorescent dye that was excited with a blue light-emitting diode. The emitted light was observed by eye and recorded with a cell-phone camera. When the target consisted of Anopheles gambiae, the reactor containing primers specific to An. gambiae lit up while the other two reactors remained dark. When the target consisted of Anopheles arabiensis, the reactor containing primers specific to An. arabiensis lit up while the other two reactors remained dark. CONCLUSIONS/SIGNIFICANCE: The microfluidic chip provides a means to identify mosquito type through molecular analysis. It is suitable for field work, allowing one to

  6. 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.

  7. 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

  8. SNP discovery and molecular evolution in Anopheles gambiae, with special emphasis on innate immune system

    OpenAIRE

    Koutsos Anastasios; Morlais Isabelle; Simard Frédéric; Krishnakumar Sujatha; Cohuet Anna; Fontenille Didier; Mindrinos Michael; Kafatos Fotis C

    2008-01-01

    Abstract Background Anopheles innate immunity affects Plasmodium development and is a potential target of innovative malaria control strategies. The extent and distribution of nucleotide diversity in immunity genes might provide insights into the evolutionary forces that condition pathogen-vector interactions. The discovery of polymorphisms is an essential step towards association studies of susceptibility to infection. Results We sequenced coding fragments of 72 immune related genes in natur...

  9. Natural plant sugar sources of Anopheles mosquitoes strongly impact malaria transmission potential.

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    Weidong Gu

    Full Text Available An improved knowledge of mosquito life history could strengthen malaria vector control efforts that primarily focus on killing mosquitoes indoors using insecticide treated nets and indoor residual spraying. Natural sugar sources, usually floral nectars of plants, are a primary energy resource for adult mosquitoes but their role in regulating the dynamics of mosquito populations is unclear. To determine how the sugar availability impacts Anopheles sergentii populations, mark-release-recapture studies were conducted in two oases in Israel with either absence or presence of the local primary sugar source, flowering Acacia raddiana trees. Compared with population estimates from the sugar-rich oasis, An. sergentii in the sugar-poor oasis showed smaller population size (37,494 vs. 85,595, lower survival rates (0.72 vs. 0.93, and prolonged gonotrophic cycles (3.33 vs. 2.36 days. The estimated number of females older than the extrinsic incubation period of malaria (10 days in the sugar rich site was 4 times greater than in the sugar poor site. Sugar feeding detected in mosquito guts in the sugar-rich site was significantly higher (73% than in the sugar-poor site (48%. In contrast, plant tissue feeding (poor quality sugar source in the sugar-rich habitat was much less (0.3% than in the sugar-poor site (30%. More important, the estimated vectorial capacity, a standard measure of malaria transmission potential, was more than 250-fold higher in the sugar-rich oasis than that in the sugar-poor site. Our results convincingly show that the availability of sugar sources in the local environment is a major determinant regulating the dynamics of mosquito populations and their vector potential, suggesting that control interventions targeting sugar-feeding mosquitoes pose a promising tactic for combating transmission of malaria parasites and other pathogens.

  10. 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...

  11. 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.

  12. 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...

  13. Stereoscopic video analysis of Anopheles gambiae behavior in the field: challenges and opportunities

    Science.gov (United States)

    Advances in our ability to localize and track individual swarming mosquitoes in the field via stereoscopic image analysis have enabled us to test long standing ideas about individual male behavior and directly observe coupling. These studies further our fundamental understanding of the reproductive ...

  14. 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...

  15. Unbiased Characterization of Anopheles Mosquito Blood Meals by Targeted High-Throughput Sequencing.

    Science.gov (United States)

    Logue, Kyle; Keven, John Bosco; Cannon, Matthew V; Reimer, Lisa; Siba, Peter; Walker, Edward D; Zimmerman, Peter A; Serre, David

    2016-03-01

    Understanding mosquito host choice is important for assessing vector competence or identifying disease reservoirs. Unfortunately, the availability of an unbiased method for comprehensively evaluating the composition of insect blood meals is very limited, as most current molecular assays only test for the presence of a few pre-selected species. These approaches also have limited ability to identify the presence of multiple mammalian hosts in a single blood meal. Here, we describe a novel high-throughput sequencing method that enables analysis of 96 mosquitoes simultaneously and provides a comprehensive and quantitative perspective on the composition of each blood meal. We validated in silico that universal primers targeting the mammalian mitochondrial 16S ribosomal RNA genes (16S rRNA) should amplify more than 95% of the mammalian 16S rRNA sequences present in the NCBI nucleotide database. We applied this method to 442 female Anopheles punctulatus s. l. mosquitoes collected in Papua New Guinea (PNG). While human (52.9%), dog (15.8%) and pig (29.2%) were the most common hosts identified in our study, we also detected DNA from mice, one marsupial species and two bat species. Our analyses also revealed that 16.3% of the mosquitoes fed on more than one host. Analysis of the human mitochondrial hypervariable region I in 102 human blood meals showed that 5 (4.9%) of the mosquitoes unambiguously fed on more than one person. Overall, analysis of PNG mosquitoes illustrates the potential of this approach to identify unsuspected hosts and characterize mixed blood meals, and shows how this approach can be adapted to evaluate inter-individual variations among human blood meals. Furthermore, this approach can be applied to any disease-transmitting arthropod and can be easily customized to investigate non-mammalian host sources.

  16. Unbiased Characterization of Anopheles Mosquito Blood Meals by Targeted High-Throughput Sequencing.

    Directory of Open Access Journals (Sweden)

    Kyle Logue

    2016-03-01

    Full Text Available Understanding mosquito host choice is important for assessing vector competence or identifying disease reservoirs. Unfortunately, the availability of an unbiased method for comprehensively evaluating the composition of insect blood meals is very limited, as most current molecular assays only test for the presence of a few pre-selected species. These approaches also have limited ability to identify the presence of multiple mammalian hosts in a single blood meal. Here, we describe a novel high-throughput sequencing method that enables analysis of 96 mosquitoes simultaneously and provides a comprehensive and quantitative perspective on the composition of each blood meal. We validated in silico that universal primers targeting the mammalian mitochondrial 16S ribosomal RNA genes (16S rRNA should amplify more than 95% of the mammalian 16S rRNA sequences present in the NCBI nucleotide database. We applied this method to 442 female Anopheles punctulatus s. l. mosquitoes collected in Papua New Guinea (PNG. While human (52.9%, dog (15.8% and pig (29.2% were the most common hosts identified in our study, we also detected DNA from mice, one marsupial species and two bat species. Our analyses also revealed that 16.3% of the mosquitoes fed on more than one host. Analysis of the human mitochondrial hypervariable region I in 102 human blood meals showed that 5 (4.9% of the mosquitoes unambiguously fed on more than one person. Overall, analysis of PNG mosquitoes illustrates the potential of this approach to identify unsuspected hosts and characterize mixed blood meals, and shows how this approach can be adapted to evaluate inter-individual variations among human blood meals. Furthermore, this approach can be applied to any disease-transmitting arthropod and can be easily customized to investigate non-mammalian host sources.

  17. The molecular evolution of four anti-malarial immune genes in the Anopheles gambiae species complex

    OpenAIRE

    Simard Frederic; Antonio-Nkondjio Christophe; Awono-Ambene Parfait H; Marshall Jonathon C; Slotman Michel A; Parmakelis Aristeidis; Caccone Adalgisa; Powell Jeffrey R

    2008-01-01

    Abstract Background If the insect innate immune system is to be used as a potential blocking step in transmission of malaria, then it will require targeting one or a few genes with highest relevance and ease of manipulation. The problem is to identify and manipulate those of most importance to malaria infection without the risk of decreasing the mosquito's ability to stave off infections by microbes in general. Molecular evolution methodologies and concepts can help identify such genes. Withi...

  18. Gal4-based enhancer-trapping in the malaria mosquito Anopheles stephensi.

    Science.gov (United States)

    O'Brochta, David A; Pilitt, Kristina L; Harrell, Robert A; Aluvihare, Channa; Alford, Robert T

    2012-11-01

    Transposon-based forward and reverse genetic technologies will contribute greatly to ongoing efforts to study mosquito functional genomics. A piggyBac transposon-based enhancer-trap system was developed that functions efficiently in the human malaria vector, Anopheles stephensi. The system consists of six transgenic lines of Anopheles stephensi, each with a single piggyBac-Gal4 element in a unique genomic location; six lines with a single piggyBac-UAStdTomato element; and two lines, each with a single Minos element containing the piggyBac-transposase gene under the regulatory control of the hsp70 promoter from Drosophila melanogaster. Enhancer detection depended upon the efficient remobilization of piggyBac-Gal4 transposons, which contain the yeast transcription factor gene Gal4 under the regulatory control of a basal promoter. Gal4 expression was detected through the expression of the fluorescent protein gene tdTomato under the regulatory control of a promoter with Gal4-binding UAS elements. From five genetic screens for larval- and adult-specific enhancers, 314 progeny were recovered from 24,250 total progeny (1.3%) with unique patterns of tdTomato expression arising from the influence of an enhancer. The frequency of piggyBac remobilization and enhancer detection was 2.5- to 3-fold higher in female germ lines compared with male germ lines. A small collection of enhancer-trap lines are described in which Gal4 expression occurred in adult female salivary glands, midgut, and fat body, either singly or in combination. These three tissues play critical roles during the infection of Anopheles stephensi by malaria-causing Plasmodium parasites. This system and the lines generated using it will be valuable resources to ongoing mosquito functional genomics efforts.

  19. Studies on the bionomics of male Anopheles gambiae Giles and male Anopheles funestus Giles from southern Mozambique

    DEFF Research Database (Denmark)

    Charlwood, J D

    2011-01-01

    . gambiae s.l. rest inside houses. They leave in a dusk exodus, which makes them easy to collect. In 8,348 exit collections from a village from 2003 to 2009, 567,195 male An. funestus and 34,591 male An. gambiae s.l. were collected. During the study, numbers of An. funestus increased but numbers of An...

  20. Green tea proanthocyanidins cause impairment of hormone-regulated larval development and reproductive fitness via repression of juvenile hormone acid methyltransferase, insulin-like peptide and cytochrome P450 genes in Anopheles gambiae sensu stricto

    Science.gov (United States)

    Nyanjom, Steven G.; Mutunga, James M.; Njeru, Sospeter N.; Bargul, Joel L.

    2017-01-01

    Successful optimization of plant-derived compounds into control of nuisance insects would benefit from scientifically validated targets. However, the close association between the genotypic responses and physiological toxicity effects mediated by these compounds remains underexplored. In this study, we evaluated the sublethal dose effects of proanthocyanidins (PAs) sourced from green tea (Camellia sinensis) on life history traits of Anopheles gambiae (sensu stricto) mosquitoes with an aim to unravel the probable molecular targets. Based on the induced phenotypic effects, genes selected for study targeted juvenile hormone (JH) biosynthesis, signal transduction, oxidative stress response and xenobiotic detoxification in addition to vitellogenesis in females. Our findings suggest that chronic exposure of larval stages (L3/L4) to sublethal dose of 5 ppm dramatically extended larval developmental period for up to 12 days, slowed down pupation rates, induced abnormal larval-pupal intermediates and caused 100% inhibition of adult emergence. Further, females exhibited significant interference of fecundity and egg hatchability relative to controls (p reproductive fitness thus could be potentially used for controlling populations of malaria vectors. PMID:28301607

  1. 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.

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

    Directory of Open Access Journals (Sweden)

    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

  3. 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...

  4. Differential effect of human ivermectin treatment on blood feeding Anopheles gambiae and Culex quinquefasciatus

    OpenAIRE

    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, in an area of north-eastern Tanzania which has been exposed to ivermectin mass drug administration, this study assessed and compared the effect of human ivermectin treatment on blood feeding Anopheles ...

  5. The role of proboscis of the malaria vector mosquito Anopheles stephensi in host-seeking behavior

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    Yoshimura Aya

    2011-01-01

    Full Text Available Abstract Background The proboscis is an essential head appendage in insects that processes gustatory code during food intake, particularly useful considering that blood-sucking arthropods routinely reach vessels under the host skin using this proboscis as a probe. Results Here, using an automated device able to quantify CO2-activated thermo (35°C-sensing behavior of the malaria vector Anopheles stephensi, we uncovered that the protruding proboscis of mosquitoes contributes unexpectedly to host identification from a distance. Ablation experiments indicated that not only antennae and maxillary palps, but also proboscis were required for the identification of pseudo-thermo targets. Furthermore, the function of the proboscis during this behavior can be segregated from CO2 detection required to evoke mosquito activation, suggesting that the proboscis of mosquitoes divide the proboscis into a "thermo-antenna" in addition to a "thermo-probe". Conclusions Our findings support an emerging view with a possible role of proboscis as important equipment during host-seeking, and give us an insight into how these appendages likely evolved from a common origin in order to function as antenna organs.

  6. Production of a Transgenic Mosquito Expressing Circumsporozoite Protein, a Malarial Protein, in the Salivary Gland of Anopheles stephensi (Diptera:Culicidae)

    OpenAIRE

    Matsuoka, Hiroyuki; Ikezawa, Tsunetaka; Hirai, Makoto

    2010-01-01

    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 co...

  7. A Study of the Essential Oils of Four Sudanese Accessions of Basil (Ocimum basilicum L. Against Anopheles Mosquito Larvae

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    Azhari H. Nour

    2009-01-01

    Full Text Available Problem statement: Certain basil essential oils were claimed to have a larvicidal activity towards mosquito's larvae. To test this claim the essential oils of four accessions of basil grown in Sudan were selected and tested for Anopheles larvae. Malaria is the major health problem in the Sudan and the whole country is now considered endemic, with varying degrees, about 35,000 deaths every year due to malaria. Anopheles mosquito is the major vector of malaria disease in Sudan. Search for larvicidal active compound(s is one of several attempts to fine effective and affordable ways to control this mosquito. To determine the toxic effects of basil essential oils extracted by steam distillation against Anopheles larvae. Approach: For the larvicidal bioassay, three concentrations (100, 300, 500 ppm of essential oil solutions of four basil accessions were prepared; 1 mL of ethanol was used to solubilize the oil in water (999 mL. In each concentration of oil solution were inserted 20 larvae (third instars. A set of controls using 0.1% ethanol and untreated sets of larvae in (tap water, were also run for comparison. Data were evaluated through regression analysis, from the regression line; the LC50 values were read. The active ingredients were separated and/ or identified by TLC, IR and GC-MS. Results: Larvicidal activity of the essential oils is varied, lasted for about 9 h and thereafter decreased, LC50 values ranging from 190-300 ppm. Linalool, geraniol and eugenol are active components of basil essential oil against Anopheles larvae. Two accessions were caused 100% mortality at a concentration range 300-500 ppm for 3 h. Conclusion: These results indicated that basil essential oils have larvicidal activity towards Anopheles larvae. Therefore, could be affordable way to control this mosquito.

  8. 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

  9. Repellency effect of forty-one essential oils against Aedes, Anopheles, and Culex mosquitoes.

    Science.gov (United States)

    Amer, Abdelkrim; Mehlhorn, Heinz

    2006-09-01

    Since ancient times, plant products were used in various aspects. However, their use against pests decreased when chemical products became developed. Recently, concerns increased with respect to public health and environmental security requiring detection of natural products that may be used against insect pests. In this study, 41 plant extracts and 11 oil mixtures were evaluated against the yellow fever mosquito, Aedes aegypti (Linnaeus), the malaria vector, Anopheles stephensi (Liston), and the filariasis and encephalitis vector, Culex quinquefasciatus (Say) (Diptera: Culicidae) using the skin of human volunteers to find out the protection time and repellency. The five most effective oils were those of Litsea (Litsea cubeba), Cajeput (Melaleuca leucadendron), Niaouli (Melaleuca quinquenervia), Violet (Viola odorata), and Catnip (Nepeta cataria), which induced a protection time of 8 h at the maximum and a 100% repellency against all three species. This effect needs, however, a peculiar formulation to fix them on the human skin.

  10. Quetzal: a transposon of the Tc1 family in the mosquito Anopheles albimanus.

    Science.gov (United States)

    Ke, Z; Grossman, G L; Cornel, A J; Collins, F H

    1996-10-01

    A member of the Tc1 family of transposable elements has been identified in the Central and South American mosquito Anopheles albimanus. The full-length Quetzal element is 1680 base pairs (bp) in length, possesses 236 bp inverted terminal repeats (ITRs), and has a single open reading frame (ORF) with the potential of encoding a 341-amino-acid (aa) protein that is similar to the transposases of other members of the Tc1 family, particularly elements described from three different Drosophila species. The approximately 10-12 copies per genome of Quetzal are found in the euchromatin of all three chromosomes of A. albimanus. One full-length clone, Que27, appears capable of encoding a complete transposase and may represent a functional copy of this element.

  11. Radiation-induced sterility for pupal and adult stages of the malaria mosquito Anopheles arabiensis

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    Knols Bart GJ

    2006-05-01

    Full Text Available Abstract 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 Results Irradiation of pupae, for all doses tested, had no effect on adult emergence. Survival curves of males irradiated as pupae or adults were similar or even slightly higher than non-irradiated males. Overall, adults appeared to be slightly more susceptible to irradiation, although no significant differences for individual doses were observed. In the pupal stage, a significant negative correlation was found between insemination and dose, but the correlation-coefficient was associated with less than 25% of the total variation. A review of the literature indicated that An. arabiensis is more radiation resistant than other anopheline mosquitoes. Conclusion The optimal dose for male insects to be released in an SIT programme depends on their level of sterility and competitiveness. The use of semi-sterilizing doses to produce more competitive insects is discussed. The most convenient developmental stage for mosquito irradiation on a mass-scale are pupae, but pupal irradiation resulted in a lower insemination rate at the highest dose compared to adult irradiation. On the basis of this study, a suitable dose range that includes semi-sterilizing doses is identified to initiate competitiveness experiments for males irradiated at both developmental stages.

  12. Genotyping of chloroquine resistant Plasmodium falciparum in wild caught Anopheles minimus mosquitoes in a malaria endemic area of Assam, India.

    Science.gov (United States)

    Sarma, D K; Mohapatra, P K; Bhattacharyya, D R; Mahanta, J; Prakash, A

    2014-09-01

    We validated the feasibility of using Plasmodium falciparum, the human malaria parasite, DNA present in wild caught vector mosquitoes for the characterization of chloroquine resistance status. House frequenting mosquitoes belonging to Anopheles minimus complex were collected from human dwellings in a malaria endemic area of Assam, Northeast India and DNA was extracted from the head-thorax region of individual mosquitoes. Anopheles minimus complex mosquitoes were identified to species level and screened for the presence of Plasmodium sp. using molecular tools. Nested PCR-RFLP method was used for genotyping of P. falciparum based on K76T mutation in the chloroquine resistance transporter (pfcrt) gene. Three of the 27 wild caught An. minimus mosquitoes were harbouring P. falciparum sporozoites (positivity 11.1%) and all 3 were had 76T mutation in the pfcrt gene, indicating chloroquine resistance. The approach of characterizing antimalarial resistance of malaria parasite in vector mosquitoes can potentially be used as a surveillance tool for monitoring transmission of antimalarial drug resistant parasite strains in the community.

  13. Production of a transgenic mosquito expressing circumsporozoite protein, a malarial protein, in the salivary gland of Anopheles stephensi (Diptera: Culicidae).

    Science.gov (United States)

    Matsuoka, Hiroyuki; Ikezawa, Tsunetaka; Hirai, Makoto

    2010-08-01

    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 40 ng 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.

  14. Molecular Evolution of Immune Genes in the Malaria Mosquito Anopheles gambiae

    OpenAIRE

    Tovi Lehmann; Jen C C Hume; Monica Licht; Burns, Christopher S.; Kurt Wollenberg; Fred Simard; Ribeiro, Jose M. C.

    2009-01-01

    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 polymorphis...

  15. Screening of selected ethnomedicinal plants from South Africa for larvicidal activity against the mosquito Anopheles arabiensis

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    Maharaj Rajendra

    2012-09-01

    Full Text Available Abstract Background This study was initiated to establish whether any South African ethnomedicinal plants (indigenous or exotic, that have been reported to be used traditionally to repel or kill mosquitoes, exhibit effective mosquito larvicidal properties. Methods Extracts of a selection of plant taxa sourced in South Africa were tested for larvicidal properties in an applicable assay. Thirty 3rd instar Anopheles arabiensis larvae were exposed to various extract types (dichloromethane, dichloromethane/methanol (1:1, methanol and purified water of each species investigated. Mortality was evaluated relative to the positive control Temephos (Mostop; Agrivo, an effective emulsifiable concentrate larvicide. Results Preliminary screening of crude extracts revealed substantial variation in toxicity with 24 of the 381 samples displaying 100% larval mortality within the seven day exposure period. Four of the high activity plants were selected and subjected to bioassay guided fractionation. The results of the testing of the fractions generated identified one fraction of the plant, Toddalia asiatica as being very potent against the An. arabiensis larvae. Conclusion The present study has successfully identified a plant with superior larvicidal activity at both the crude and semi pure fractions generated through bio-assay guided fractionation. These results have initiated further research into isolating the active compound and developing a malaria vector control tool.

  16. Artificial activation of mature unfertilized eggs in the malaria vector mosquito, Anopheles stephensi (Diptera, Culicidae).

    Science.gov (United States)

    Yamamoto, Daisuke S; Hatakeyama, Masatsugu; Matsuoka, Hiroyuki

    2013-08-01

    In the past decade, many transgenic lines of mosquitoes have been generated and analyzed, whereas the maintenance of a large number of transgenic lines requires a great deal of effort and cost. In vitro fertilization by an injection of cryopreserved sperm into eggs has been proven to be effective for the maintenance of strains in mammals. The technique of artificial egg activation is a prerequisite for the establishment of in vitro fertilization by sperm injection. We demonstrated that artificial egg activation is feasible in the malaria vector mosquito, Anopheles stephensi (Diptera, Culicidae). Nearly 100% of eggs dissected from virgin females immersed in distilled water darkened, similar to normally oviposited fertilized eggs. It was revealed by the cytological examination of chromosomes that meiotic arrest was relieved in these eggs approximately 20 min after incubation in water. Biochemical examinations revealed that MAPK (mitogen-activated protein kinase)/ERK (extracellular signal-regulated protein kinase) and MEK (MAPK/ERK kinase) were dephosphorylated similar to that in fertilized eggs. These results indicate that dissected unfertilized eggs were activated in distilled water and started development. Injection of distilled water into body cavity of the virgin blood-fed females also induced activation of a portion of eggs in the ovaries. The technique of artificial egg activation is expected to contribute to the success of in vitro fertilization in A. stephensi.

  17. Laser dosimetry for disabling anopheles stephensi mosquitoes in-flight (Conference Presentation)

    Science.gov (United States)

    Keller, Matthew D.; Norton, Bryan J.; Rutschman, Phil; Farrar, David J.; Marvit, Maclen; Makagon, Artyom

    2016-03-01

    The Photonic Fence is a system designed to detect mosquitoes and other pestilent flying insects in an active region and to apply lethal doses of laser light to them. Previously, we determined lethal fluence levels for a variety of lasers and pulse conditions on anesthetized Anopheles stephensi mosquitoes. In this work, similar studies were performed while the bugs were freely flying within transparent cages. Dose-response curves were created for various beam diameter, pulse width, and power conditions at 455 nm, 532 nm, 1064nm, and 1540 nm wavelengths. Besides mortality outcomes, the flight behavior of the bugs and the performance of the tracking system were monitored for consistency and to ensure that they had no impact on the mortality outcomes. As in anesthetized experiments, the visible wavelengths required significantly less fluence than near infrared wavelengths to reliably disable bugs. For the visible wavelengths, lethal fluence values were generally equivalent to those found in anesthetized dosing, while near infrared wavelengths required approximately twice the fluence compared with anesthetized experiments. The performance of the optical tracking system remained highly stable throughout the experiments, and it was found not to influence mortality results for pulse widths up to 25 ms. In general, keeping energy constant while decreasing power and increasing pulse width reduced mortality levels. The results of this study further affirm the practicality of using optical approaches to protect people and crops from flying insects.

  18. Mathematical evaluation of community level impact of combining bed nets and indoor residual spraying upon malaria transmission in areas where the main vectors are Anopheles arabiensis mosquitoes

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    Okumu Fredros O

    2013-01-01

    Full Text Available Abstract Background Indoor residual insecticide spraying (IRS and long-lasting insecticide treated nets (LLINs are commonly used together even though evidence that such combinations confer greater protection against malaria than either method alone is inconsistent. Methods A deterministic model of mosquito life cycle processes was adapted to allow parameterization with results from experimental hut trials of various combinations of untreated nets or LLINs (Olyset®, PermaNet 2.0®, Icon Life® nets with IRS (pirimiphos methyl, lambda cyhalothrin, DDT, in a setting where vector populations are dominated by Anopheles arabiensis, so that community level impact upon malaria transmission at high coverage could be predicted. Results Intact untreated nets alone provide equivalent personal protection to all three LLINs. Relative to IRS plus untreated nets, community level protection is slightly higher when Olyset® or PermaNet 2.0® nets are added onto IRS with pirimiphos methyl or lambda cyhalothrin but not DDT, and when Icon Life® nets supplement any of the IRS insecticides. Adding IRS onto any net modestly enhances communal protection when pirimiphos methyl is sprayed, while spraying lambda cyhalothrin enhances protection for untreated nets but not LLINs. Addition of DDT reduces communal protection when added to LLINs. Conclusions Where transmission is mediated primarily by An. arabiensis, adding IRS to high LLIN coverage provides only modest incremental benefit (e.g. when an organophosphate like pirimiphos methyl is used, but can be redundant (e.g. when a pyrethroid like lambda cyhalothin is used or even regressive (e.g. when DDT is used for the IRS. Relative to IRS plus untreated nets, supplementing IRS with LLINs will only modestly improve community protection. Beyond the physical protection that intact nets provide, additional protection against transmission by An. arabiensis conferred by insecticides will be remarkably small, regardless of

  19. Larvicidal properties of two asclepiadaceous plant species against the mosquito Anopheles arabiensis Patton (Diptera: Culicidae

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    Amal Elsayed Edriss

    2013-01-01

    Full Text Available Certain mosquito species are important vectors of fatal human diseases, among which Anopheles arabiensis is known to be associated with malaria transmission in different tropical and subtropical areas. Since chemical control of mosquitoes was linked with numerous drawbacks, like resistance development, the search for effective environmentally sound alternatives is urgently needed. Therefore, it was aimed by this study to evaluate some extracts prepared from two asclepiadaceous plants, viz., Solenostemma argel “Hargel” (seeds and leaves and Calotropis procera “Usher” (leaves and flowers, as natural larvicides against An. arabiensis. The main parameters included bioassays of treatments for knockdown and residual effects, besides phytochemical analysis of the tested extracts. The results revealed variable groups of secondary metabolites in the two plants, with S. argel seemed to be the richest one. Hence, S. argel extracts caused higher larval mortalities than those of C. procera. This could be ascribed to some potent secondary metabolites in the former plant, which needs further studies. Almost all the high concentrations of S. argel extracts exerted the highest knockdown effect (90% mortality after 24 h, which were comparable with those obtained by two standard insecticides. The highest doses of petroleum ether and water extracts of this plant also manifested significantly higher residual effects than the other extracts after three days following treatments, but were surpassed by the chemical insecticides thereafter. However, S. argel seed petroleum ether extract at 0.5% was the most effective of all botanicals up to three weeks of exposure. This extract needs to be evaluated under field conditions for proper exploitation as mosquito larvicide.

  20. 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

  1. Factors affecting transmission of Wuchereria bancrofti by anopheline mosquitoes. 2. Damage to ingested microfilariae by mosquito foregut armatures and development of filarial larvae in mosquitoes.

    Science.gov (United States)

    Bryan, J H; Southgate, B A

    1988-01-01

    Microfilariae (mf) of Wuchereria bancrofti from the midgut of 639 Anopheles gambiae, 557 An. arabiensis, 117 An. melas and 9 An. funestus were examined immediately after the mosquitoes had fed on carriers with different densities of mf. The percentages of mf damaged during ingestion were 57.1-60.0 in An. gambiae, 33.3-50.6 in An. arabiensis and 38.7-55.7 in An. melas. In each species the percentage of mf damaged was independent of mf density in the human host. A further 3657 An. gambiae, 2875 An. arabiensis, 347 An. melas and 32 An. funestus were examined 7 d or more after feeding on mf carriers. In An. gambiae and An. arabiensis, mean numbers of larvae per mosquito were strongly correlated to mf blood density, with similar regression slopes to those obtained from the regression of mf blood density on mean uptake of mf/mosquito. The ratio of mean numbers of larvae per mosquito to mean numbers of intact mf ingested per mosquito increased as the density of mf in the human host increased in An. gambiae and An. arabiensis, but decreased in An. melas as host mf density increased.

  2. The human malaria parasite Pfs47 gene mediates evasion of the mosquito immune system.

    Science.gov (United States)

    Molina-Cruz, Alvaro; Garver, Lindsey S; Alabaster, Amy; Bangiolo, Lois; Haile, Ashley; Winikor, Jared; Ortega, Corrie; van Schaijk, Ben C L; Sauerwein, Robert W; Taylor-Salmon, Emma; Barillas-Mury, Carolina

    2013-05-24

    Plasmodium falciparum transmission by Anopheles gambiae mosquitoes is remarkably efficient, resulting in a very high prevalence of human malaria infection in sub-Saharan Africa. A combination of genetic mapping, linkage group selection, and functional genomics was used to identify Pfs47 as a P. falciparum gene that allows the parasite to infect A. gambiae without activating the mosquito immune system. Disruption of Pfs47 greatly reduced parasite survival in the mosquito, and this phenotype could be reverted by genetic complementation of the parasite or by disruption of the mosquito complement-like system. Pfs47 suppresses midgut nitration responses that are critical to activate the complement-like system. We provide direct experimental evidence that immune evasion mediated by Pfs47 is critical for efficient human malaria transmission by A. gambiae.

  3. Attractiveness of volatiles from different body parts to the malaria mosquito Anopheles coluzzii is affected by deodorant compounds.

    Science.gov (United States)

    Verhulst, Niels O; Weldegergis, Berhane T; Menger, David; Takken, Willem

    2016-06-01

    Mosquitoes display biting preferences among different sites of the human body. In addition to height or convection currents, body odour may play a role in the selection of these biting sites. Previous studies have shown that skin emanations are important host-finding cues for mosquitoes. In this study, skin emanations were collected from armpits, hands and feet; the volatile profiles were analysed and tested for their attractiveness to the malaria mosquito Anopheles coluzzii. Skin emanations collected from armpits were less attractive to An. coluzzii compared to hands or/and feet. The difference may have been caused by deodorant residues, which were found in the armpit samples and not in those of hands and feet. In a subsequent experiment, volunteers were asked to avoid using skincare products for five days, and thereafter, no differences in attractiveness of the body parts to mosquitoes were found. The detected deodorant compound isopropyl tetradecanoate inhibited mosquito landings in a repellent bioassay. It is concluded that the volatiles emanated from different body parts induced comparable levels of attraction in mosquitoes, and that skincare products may reduce a person's attractiveness to mosquitoes.

  4. 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

  5. Mosquito larvicidal, ovicidal, and repellent properties of botanical extracts against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae).

    Science.gov (United States)

    Govindarajan, M; Mathivanan, T; Elumalai, K; Krishnappa, K; Anandan, A

    2011-08-01

    Mosquito-borne diseases have an economic impact, including loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates; however, no part of the world is free from vector-borne diseases. In mosquito control programs, botanical origin may have the potential to be used successfully as eggs, larvae, and adult. The larvicidal, ovicidal, and repellent activities of crude benzene and ethyl acetate extracts of leaf of Ervatamia coronaria and Caesalpinia pulcherrima were assayed for their toxicity against three important vector mosquitoes, viz., Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in benzene extract of E. coronaria against the larvae of Anopheles Stephensi, Aedes aegypti, and Culex quinquefasciatus with the LC(50) and LC(90) values were 79.08, 89.59, and 96.15 ppm and 150.47, 166.04, and 174.10 ppm, respectively. Mean percent hatchability of the ovicidal activity was observed 48 h posttreatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. The leaf extract of E. coronaria was found to be most effective than Caesalpinia pulcherrima against eggs/egg rafts of three vector mosquitoes. For E. coronaria, the benzene extract exerted 300, 250, and 200 ppm against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus, respectively. The results of the repellent activity of benzene and ethyl acetate extract of E. coronaria and Caesalpinia pulcherrima plants at three different concentrations of 1.0, 2.5, and 5.0 mg/cm(2) were applied on skin of fore arm in man and exposed against adult female mosquitoes. In this observation, these two plant crude extracts gave protection against mosquito bites without any allergic reaction to the test person, and also, the

  6. Insecticide resistance in Bemisia tabaci Gennadius (Homoptera: Aleyrodidae) and Anopheles gambiae Giles (Diptera: Culicidae) could compromise the sustainability of malaria vector control strategies in West Africa.

    Science.gov (United States)

    Gnankiné, Olivier; Bassolé, Imael H N; Chandre, Fabrice; Glitho, Isabelle; Akogbeto, Martin; Dabiré, Roch K; Martin, Thibaud

    2013-10-01

    Insecticides from the organophosphate (OP) and pyrethroid (PY) chemical families, have respectively, been in use for 50 and 30 years in West Africa, mainly against agricultural pests, but also against vectors of human disease. The selection pressure, with practically the same molecules year after year (mainly on cotton), has caused insecticide resistance in pest populations such as Bemisia tabaci, vector of harmful phytoviruses on vegetables. The evolution toward insecticide resistance in malaria vectors such as Anopheles gambiae sensus lato (s.l.) is probably related to the current use of these insecticides in agriculture. Thus, successful pest and vector control in West Africa requires an investigation of insect susceptibility, in relation to the identification of species and sub species, such as molecular forms or biotypes. Identification of knock down resistance (kdr) and acetylcholinesterase gene (Ace1) mutations modifying insecticide targets in individual insects and measure of enzymes activity typically involved in insecticide metabolism (oxidase, esterase and glutathion-S-transferase) are indispensable in understanding the mechanisms of resistance. Insecticide resistance is a good example in which genotype-phenotype links have been made successfully. Insecticides used in agriculture continue to select new resistant populations of B. tabaci that could be from different biotype vectors of plant viruses. As well, the evolution of insecticide resistance in An. gambiae threatens the management of malaria vectors in West Africa. It raises the question of priority in the use of insecticides in health and/or agriculture, and more generally, the question of sustainability of crop protection and vector control strategies in the region. Here, we review the susceptibility tests, biochemical and molecular assays data for B. tabaci, a major pest in cotton and vegetable crops, and An. gambiae, main vector of malaria. The data reviewed was collected in Benin and Burkina

  7. Malaria infected mosquitoes express enhanced attraction to human odor.

    Directory of Open Access Journals (Sweden)

    Renate C Smallegange

    Full Text Available There is much evidence that some pathogens manipulate the behaviour of their mosquito hosts to enhance pathogen transmission. However, it is unknown whether this phenomenon exists in the interaction of Anopheles gambiae sensu stricto with the malaria parasite, Plasmodium falciparum--one of the most important interactions in the context of humanity, with malaria causing over 200 million human cases and over 770 thousand deaths each year. Here we demonstrate, for the first time, that infection with P. falciparum causes alterations in behavioural responses to host-derived olfactory stimuli in host-seeking female An. gambiae s.s. mosquitoes. In behavioural experiments we showed that P. falciparum-infected An. gambiae mosquitoes were significantly more attracted to human odors than uninfected mosquitoes. Both P. falciparum-infected and uninfected mosquitoes landed significantly more on a substrate emanating human skin odor compared to a clean substrate. However, significantly more infected mosquitoes landed and probed on a substrate emanating human skin odor than uninfected mosquitoes. This is the first demonstration of a change of An. gambiae behaviour in response to olfactory stimuli caused by infection with P. falciparum. The results of our study provide vital information that could be used to provide better predictions of how malaria is transmitted from human being to human being by An. gambiae s.s. females. Additionally, it highlights the urgent need to investigate this interaction further to determine the olfactory mechanisms that underlie the differential behavioural responses. In doing so, new attractive compounds could be identified which could be used to develop improved mosquito traps for surveillance or trapping programmes that may even specifically target P. falciparum-infected An. gambiae s.s. females.

  8. Suboptimal Larval Habitats Modulate Oviposition of the Malaria Vector Mosquito Anopheles coluzzii.

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    Eunho Suh

    Full Text Available Selection of oviposition sites by gravid females is a critical behavioral step in the reproductive cycle of Anopheles coluzzii, which is one of the principal Afrotropical malaria vector mosquitoes. Several studies suggest this decision is mediated by semiochemicals associated with potential oviposition sites. To better understand the chemosensory basis of this behavior and identify compounds that can modulate oviposition, we examined the generally held hypothesis that suboptimal larval habitats give rise to semiochemicals that negatively influence the oviposition preference of gravid females. Dual-choice bioassays indicated that oviposition sites conditioned in this manner do indeed foster significant and concentration dependent aversive effects on the oviposition site selection of gravid females. Headspace analyses derived from aversive habitats consistently noted the presence of dimethyl disulfide (DMDS, dimethyl trisulfide (DMTS and 6-methyl-5-hepten-2-one (sulcatone each of which unitarily affected An. coluzzii oviposition preference. Electrophysiological assays across the antennae, maxillary palp, and labellum of gravid An. coluzzii revealed differential responses to these semiochemicals. Taken together, these findings validate the hypothesis in question and suggest that suboptimal environments for An. coluzzii larval development results in the release of DMDS, DMTS and sulcatone that impact the response valence of gravid females.

  9. Modeling of Anopheles minimus Mosquito NADPH-Cytochrome P450 Oxidoreductase (CYPOR and Mutagenesis Analysis

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    Pornpimol Rongnoparut

    2013-01-01

    Full Text Available Malaria is one of the most dangerous mosquito-borne diseases in many tropical countries, including Thailand. Studies in a deltamethrin resistant strain of Anopheles minimus mosquito, suggest cytochrome P450 enzymes contribute to the detoxification of pyrethroid insecticides. Purified A. minimus CYPOR enzyme (AnCYPOR, which is the redox partner of cytochrome P450s, loses flavin-adenosine di-nucleotide (FAD and FLAVIN mono-nucleotide (FMN cofactors that affect its enzyme activity. Replacement of leucine residues at positions 86 and 219 with phenylalanines in FMN binding domain increases FMN binding, enzyme stability, and cytochrome c reduction activity. Membrane-Bound L86F/L219F-AnCYPOR increases A. minimus P450-mediated pyrethroid metabolism in vitro. In this study, we constructed a comparative model structure of AnCYPOR using a rat CYPOR structure as a template. Overall model structure is similar to rat CYPOR, with some prominent differences. Based on primary sequence and structural analysis of rat and A. minimus CYPOR, C427R, W678A, and W678H mutations were generated together with L86F/L219F resulting in three soluble Δ55 triple mutants. The C427R triple AnCYPOR mutant retained a higher amount of FAD binding and increased cytochrome c reduction activity compared to wild-type and L86F/L219F-Δ55AnCYPOR double mutant. However W678A and W678H mutations did not increase FAD and NAD(PH bindings. The L86F/L219F double and C427R triple membrane-bound AnCYPOR mutants supported benzyloxyresorufin O-deakylation (BROD mediated by mosquito CYP6AA3 with a two- to three-fold increase in efficiency over wild-type AnCYPOR. The use of rat CYPOR in place of AnCYPOR most efficiently supported CYP6AA3-mediated BROD compared to all AnCYPORs.

  10. Mosquito Repellent Activity and Phytochemical Characterization of Essential Oils From Striga hermonthica, Hyptis spicigera and Ocimum basilicum Leaf Extracts

    OpenAIRE

    Gabi Baba; A.O. Lawal; Hauwa B. Sharif

    2012-01-01

    The main aim of this study is to screen the phytochemicals and compare the mosquito repellent activities of essential oils from Hyptis spicigera, Striga hermonthica and Ocimum basilicum (Basil) against Anopheles gambiae and Culex quinquefasciatus under laboratory conditions. The global threat of malaria to human race and the need to control its advances is on the focus. Mosquito is the target being the primary host in the spread of malaria. Alkaloids, saponnins, steroids, tannins and terpenoi...

  11. Effects of Anti-Mosquito Salivary Glands and Deglycosylated Midgut Antibodies of Anopheles stephensi on Fecundity and Longevity

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    H Mohammadzadeh Hajipirloo

    2005-09-01

    Full Text Available With the aim of controlling malaria by reducing vector population, the effects of antibodies produced against salivary glands and deglycosylated midgut antigens of Anopheles stephensi mosquitoes on fecundity and longevity of the same species were tested. Three deglycosylated preparations of midgut and two preparations of salivary glands were produced, conjugated with aluminum hydroxide gel, and subcutaneously injected to shoulders of TO (Turner Out-bred mice. After 4 immunizations and assurance of enough antibody production against utilized antigenic suspensions, effects of blood feeding on immunized and control mice were assayed. Insoluble preparation of midgut showed the strongest effect with 23.5% reduction in egg laying, and increasing death rate of vectors in third day after feeding. No significant reduction in fecundity or survivorship was seen with other preparations. Anopheles midgut insoluble antigens are potential candidates for designing vaccines against malaria vectors and further investigations need to be done to find effective antigens and the best way of their use.

  12. Plasmodium falciparum malaria challenge by the bite of aseptic Anopheles stephensi mosquitoes: results of a randomized infectivity trial.

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    Kirsten E Lyke

    Full Text Available BACKGROUND: Experimental infection of malaria-naïve volunteers by the bite of Plasmodium falciparum-infected mosquitoes is a preferred means to test the protective effect of malaria vaccines and drugs. The standard model relies on the bite of five infected mosquitoes to induce malaria. We examined the efficacy of malaria transmission using mosquitoes raised aseptically in compliance with current Good Manufacturing Practices (cGMPs. METHODS AND FINDINGS: Eighteen adults aged 18-40 years were randomized to receive 1, 3 or 5 bites of Anopheles stephensi mosquitoes infected with the chloroquine-sensitive NF54 strain of P. falciparum. Seventeen participants developed malaria; fourteen occurring on Day 11. The mean prepatent period was 10.9 days (9-12 days. The geometric mean parasitemia was 15.7 parasites/µL (range: 4-70 by microscopy. Polymerase chain reaction (PCR detected parasites 3.1 (range: 0-4 days prior to microscopy. The geometric mean sporozoite load was 16,753 sporozoites per infected mosquito (range: 1,000-57,500. A 1-bite participant withdrew from the study on Day 13 post-challenge and was PCR and smear negative. CONCLUSIONS: The use of aseptic, cGMP-compliant P. falciparum-infected mosquitoes is safe, is associated with a precise prepatent period compared to the standard model and appears more efficient than the standard approach, as it led to infection in 100% (6/6 of volunteers exposed to three mosquito bites and 83% (5/6 of volunteers exposed to one mosquito bite. TRIAL REGISTRATION: ClinicalTrials.gov NCT00744133.

  13. Targeted mutagenesis in the malaria mosquito using TALE nucleases.

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    Andrea L Smidler

    Full Text Available Anopheles gambiae, the main mosquito vector of human malaria, is a challenging organism to manipulate genetically. As a consequence, reverse genetics studies in this disease vector have been largely limited to RNA interference experiments. Here, we report the targeted disruption of the immunity gene TEP1 using transgenic expression of Transcription-Activator Like Effector Nucleases (TALENs, and the isolation of several TEP1 mutant A. gambiae lines. These mutations inhibited protein production and rendered TEP1 mutants hypersusceptible to Plasmodium berghei. The TALEN technology opens up new avenues for genetic analysis in this disease vector and may offer novel biotechnology-based approaches for malaria control.

  14. Targeted mutagenesis in the malaria mosquito using TALE nucleases.

    Science.gov (United States)

    Smidler, Andrea L; Terenzi, Olivier; Soichot, Julien; Levashina, Elena A; Marois, Eric

    2013-01-01

    Anopheles gambiae, the main mosquito vector of human malaria, is a challenging organism to manipulate genetically. As a consequence, reverse genetics studies in this disease vector have been largely limited to RNA interference experiments. Here, we report the targeted disruption of the immunity gene TEP1 using transgenic expression of Transcription-Activator Like Effector Nucleases (TALENs), and the isolation of several TEP1 mutant A. gambiae lines. These mutations inhibited protein production and rendered TEP1 mutants hypersusceptible to Plasmodium berghei. The TALEN technology opens up new avenues for genetic analysis in this disease vector and may offer novel biotechnology-based approaches for malaria control.

  15. Maternal germline-specific genes in the Asian malaria mosquito Anopheles stephensi: characterization and application for disease control.

    Science.gov (United States)

    Biedler, James K; Qi, Yumin; Pledger, David; James, Anthony A; Tu, Zhijian

    2014-12-05

    Anopheles stephensi is a principal vector of urban malaria on the Indian subcontinent and an emerging model for molecular and genetic studies of mosquito biology. To enhance our understanding of female mosquito reproduction, and to develop new tools for basic research and for genetic strategies to control mosquito-borne infectious diseases, we identified 79 genes that displayed previtellogenic germline-specific expression based on RNA-Seq data generated from 11 life stage-specific and sex-specific samples. Analysis of this gene set provided insights into the biology and evolution of female reproduction. Promoters from two of these candidates, vitellogenin receptor and nanos, were used in independent transgenic cassettes for the expression of artificial microRNAs against suspected mosquito maternal-effect genes, discontinuous actin hexagon and myd88. We show these promoters have early germline-specific expression and demonstrate 73% and 42% knockdown of myd88 and discontinuous actin hexagon mRNA in ovaries 48 hr after blood meal, respectively. Additionally, we demonstrate maternal-specific delivery of mRNA and protein to progeny embryos. We discuss the application of this system of maternal delivery of mRNA/miRNA/protein in research on mosquito reproduction and embryonic development, and for the development of a gene drive system based on maternal-effect dominant embryonic arrest.

  16. The Genome of Anopheles darlingi, the main neotropical malaria vector

    Science.gov (United States)

    Marinotti, Osvaldo; Cerqueira, Gustavo C.; de Almeida, Luiz Gonzaga Paula; Ferro, Maria Inês Tiraboschi; Loreto, Elgion Lucio da Silva; Zaha, Arnaldo; Teixeira, Santuza M. R.; Wespiser, Adam R.; Almeida e Silva, Alexandre; Schlindwein, Aline Daiane; Pacheco, Ana Carolina Landim; da Silva, Artur Luiz da Costa; Graveley, Brenton R.; Walenz, Brian P.; Lima, Bruna de Araujo; Ribeiro, Carlos Alexandre Gomes; Nunes-Silva, Carlos Gustavo; de Carvalho, Carlos Roberto; Soares, Célia Maria de Almeida; de Menezes, Claudia Beatriz Afonso; Matiolli, Cleverson; Caffrey, Daniel; Araújo, Demetrius Antonio M.; de Oliveira, Diana Magalhães; Golenbock, Douglas; Grisard, Edmundo Carlos; Fantinatti-Garboggini, Fabiana; de Carvalho, Fabíola Marques; Barcellos, Fernando Gomes; Prosdocimi, Francisco; May, Gemma; de Azevedo Junior, Gilson Martins; Guimarães, Giselle Moura; Goldman, Gustavo Henrique; Padilha, Itácio Q. M.; Batista, Jacqueline da Silva; Ferro, Jesus Aparecido; Ribeiro, José M. C.; Fietto, Juliana Lopes Rangel; Dabbas, Karina Maia; Cerdeira, Louise; Agnez-Lima, Lucymara Fassarella; Brocchi, Marcelo; de Carvalho, Marcos Oliveira; Teixeira, Marcus de Melo; Diniz Maia, Maria de Mascena; Goldman, Maria Helena S.; Cruz Schneider, Maria Paula; Felipe, Maria Sueli Soares; Hungria, Mariangela; Nicolás, Marisa Fabiana; Pereira, Maristela; Montes, Martín Alejandro; Cantão, Maurício E.; Vincentz, Michel; Rafael, Miriam Silva; Silverman, Neal; Stoco, Patrícia Hermes; Souza, Rangel Celso; Vicentini, Renato; Gazzinelli, Ricardo Tostes; Neves, Rogério de Oliveira; Silva, Rosane; Astolfi-Filho, Spartaco; Maciel, Talles Eduardo Ferreira; Ürményi, Turán P.; Tadei, Wanderli Pedro; Camargo, Erney Plessmann; de Vasconcelos, Ana Tereza Ribeiro

    2013-01-01

    Anopheles darlingi is the principal neotropical malaria vector, responsible for more than a million cases of malaria per year on the American continent. Anopheles darlingi diverged from the African and Asian malaria vectors ∼100 million years ago (mya) and successfully adapted to the New World environment. Here we present an annotated reference A. darlingi genome, sequenced from a wild population of males and females collected in the Brazilian Amazon. A total of 10 481 predicted protein-coding genes were annotated, 72% of which have their closest counterpart in Anopheles gambiae and 21% have highest similarity with other mosquito species. In spite of a long period of divergent evolution, conserved gene synteny was observed between A. darlingi and A. gambiae. More than 10 million single nucleotide polymorphisms and short indels with potential use as genetic markers were identified. Transposable elements correspond to 2.3% of the A. darlingi genome. Genes associated with hematophagy, immunity and insecticide resistance, directly involved in vector–human and vector–parasite interactions, were identified and discussed. This study represents the first effort to sequence the genome of a neotropical malaria vector, and opens a new window through which we can contemplate the evolutionary history of anopheline mosquitoes. It also provides valuable information that may lead to novel strategies to reduce malaria transmission on the South American continent. The A. darlingi genome is accessible at www.labinfo.lncc.br/index.php/anopheles-darlingi. PMID:23761445

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

    NARCIS (Netherlands)

    Howard, A.F.V.; Koenraadt, C.J.M.; Farenhorst, M.; Knols, B.G.J.; Takken, W.

    2010-01-01

    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 pr

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

    NARCIS (Netherlands)

    Howard, A.F.V.; Koenraadt, C.J.M.; Farenhorst, M.; Knols, B.G.J.; Takken, W.

    2010-01-01

    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 pr

  19. Effect of Novaluron (Rimon 10 EC) on the mosquitoes Anopheles albimanus, Anopheles pseudopunctipennis, Aedes aegypti, Aedes albopictus and Culex quinquefasciatus from Chiapas, Mexico.

    Science.gov (United States)

    Arredondo-Jiménez, J I; Valdez-Delgado, K M

    2006-12-01

    Dengue fever is a serious problem in Mexico and vector control has not been effective enough at preventing outbreaks. Malaria is largely under control, but it is important that new control measures continue to be developed. Novaluron, a novel host-specific insect growth regulator and chitin synthesis inhibitor, has proved to be effective against agricultural pests, but its efficacy against larval mosquito vectors under field conditions remains unknown. In accordance with the World Health Organization Pesticide Evaluation Scheme, phase I, II and III studies were conducted to evaluate the efficacy and residual effect of Novaluron (Rimon 10 EC, Makhteshim, Beer-Sheva, Israel) on the malaria vectors Anopheles albimanus Wiedemann (Diptera: Culicidae) and Anopheles pseudopunctipennis Theobald, the dengue vectors Aedes aegypti (L) and Aedes albopictus Skuse and the nuisance mosquito Culex quinquefasciatus Say. Laboratory susceptibility tests yielded diagnostic concentrations for all five target species. Field trials to identify the optimum field dosage of Novaluron against Anopheles mosquitoes were carried out under semi-natural conditions in artificial plots and in vessels with wild mosquitoes. Efficacy was measured by monitoring mortality of larvae and pupae and the percentage of inhibition of emergence from floating cages. Dosages of Novaluron for field tests were based on pupal LC(99) (lethal concentration 99%) of An. pseudopunctipennis (0.166 mg/L) in plots and average pupal LC(99) of Ae. aegypti and Ae. albopictus (0.55 mg/L). At all dosages tested, Novaluron significantly reduced larval populations of An. albimanus, Culex coronator Dyar & Knab, Ae. albopictus and Cx. quinquefasciatus by approximately 90%, inhibited adult emergence of An. albimanus and An. pseudopunctipennis by approximately 97% for almost 4 months in experimental plots, and inhibited adult emergence of Ae. aegypti and Ae. albopictus by approximately 97% for up to 14 weeks. Recommended dosages of

  20. Single concentration tests show synergism among Bacillus thuringiensis subsp. israelensis toxins against the malaria vector mosquito Anopheles albimanus.

    Science.gov (United States)

    Fernández-Luna, María Teresa; Tabashnik, Bruce E; Lanz-Mendoza, Humberto; Bravo, Alejandra; Soberón, Mario; Miranda-Ríos, Juan

    2010-07-01

    Bioassays of insecticidal proteins from Bacillus thuringiensis subsp. israelensis with larvae of the malaria vector mosquito Anopheles albimanus showed that the cytolytic protein Cyt1Aa was not toxic alone, but it increased the toxicity of the crystalline proteins Cry4Ba and Cry11Aa. Synergism also occurred between Cry4Ba and Cry11Aa toxins. Whereas many previous analyses of synergism have been based on a series of toxin concentrations leading to comparisons between expected and observed values for the concentration killing 50% of insects tested (LC(50)), we describe and apply a method here that enables testing for synergism based on single concentrations of toxins.

  1. Larvicidal property of green synthesized silver nanoparticles against vector mosquitoes (Anopheles stephensi and Aedes aegypti

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    Rajamani Bhuvaneswari

    2016-10-01

    Full Text Available Mosquito vectors spread severe human diseases which lead to millions of deaths every year. Vector management is ultimately aimed to develop the health of every individual’s life by reducing the mosquito diversity. Control of vectors in growing counties is an important issue with various aspects. The advancement of green nanotechnology will attribute the solution for vector controlling policy. To identify the larvicidal property of silver nanoparticles (AgNPs using Belosynapsis kewensis (B. kewensis leaf extract against the Anopheles stephensi (A. stephensi and Aedes aegypti (A. aegypti in vitro study (LC50 and LC90 was analyzed. The synthesized AgNPs were characterized by UV–vis. (Ultraviolet–visible spectroscopy, FTIR (Fourier Transform Infra Red spectroscopy, TEM (Transmission Electron Microscopy, and XRD (X-ray Diffraction. Green AgNPs have a maximum absorption at 411 nm. The FTIR spectrum showed prominent peaks in (3863.55, 3759.02, 3361.01, 2926.81, 1575.12, 1388.16, 1034.79, 821.96, 717.07, 590.92, 534.32 and 472.42 cm−1 in the region of 4000–400 cm−1. The XRD peaks shown at 27.4°, 35.90°, 37.20°, 51.23° and 71.10° correspond to (311, (100, (101, (104 and (006 planes for face centered cubic (FCC. The TEM image showed the NPs with an average size of 24 ± 1.6 nm. In vitro larvicidal activity of AgNPs was used against the fourth instar of A. stephensi and A. aegypti. The LC50 and LC90 values of AgNPs showed to be effective against A. stephensi (LC50 = 78.4; LC90 = 144.7 ppm followed by A. aegypti (LC50 = 84.2; LC90 = 117.3 ppm. These results recommend that the green synthesized AgNPs have a potential to be used as a candidate for the control of A. stephensi and A. aegypti through eco-friendly and cost effective approach.

  2. [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.

  3. 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...

  4. Bitter-sensitive gustatory receptor neuron responds to chemically diverse insect repellents in the common malaria mosquito Anopheles quadrimaculatus

    Science.gov (United States)

    Sparks, Jackson T.; Dickens, Joseph C.

    2016-06-01

    Female mosquitoes feed on blood from animal hosts to obtain nutritional resources used for egg production. These contacts facilitate the spread of harmful human diseases. Chemical repellents are used to disrupt mosquito host-seeking and blood-feeding behaviors; however, little is known about the gustatory sensitivity of mosquitoes to known repellents. Here, we recorded electrical responses from gustatory receptor neurons (GRNs) housed within the labellar sensilla of female Anopheles quadrimaculatus to N,N-diethyl-3-methylbenzamide (DEET), picaridin, IR3535, 2-undecanone, p-menthane-3,8-diol, geraniol, trans-2-hexen-1-ol, quinine, and quinidine. A bitter-sensitive GRN responded to all tested repellents and quinine, a known feeding deterrent. Responses of the bitter-sensitive neuron to quinine and an isomer, quinidine, did not differ. Delayed bursts of electrical activity were observed in response to continuous stimulation with synthetic repellents at high concentrations. Electrophysiological recordings from bitter-sensitive GRNs associated with mosquito gustatory sensilla represent a convenient model to evaluate candidate repellents.

  5. Tissue-specific features of the X chromosome and nucleolus spatial dynamics in a malaria mosquito, Anopheles atroparvus

    Science.gov (United States)

    Bondarenko, Semen M.; Artemov, Gleb N.; Stegniy, Vladimir N.

    2017-01-01

    Spatial organization of chromosome territories is important for maintenance of genomic stability and regulation of gene expression. Recent studies have shown tissue-specific features of chromosome attachments to the nuclear envelope in various organisms including malaria mosquitoes. However, other spatial characteristics of nucleus organization, like volume and shape of chromosome territories, have not been studied in Anopheles. We conducted a thorough analysis of tissue-specific features of the X chromosome and nucleolus volume and shape in follicular epithelium and nurse cells of the Anopheles atroparvus ovaries using a modern open-source software. DNA of the polytene X chromosome from ovarian nurse cells was obtained by microdissection and was used as a template for amplification with degenerate oligo primers. A fluorescently labeled X chromosome painting probe was hybridized with formaldehyde-fixed ovaries of mosquitoes using a 3D-FISH method. The nucleolus was stained by immunostaining with an anti-fibrillarin antibody. The analysis was conducted with TANGO—a software for a chromosome spatial organization analysis. We show that the volume and position of the X chromosome have tissue-specific characteristics. Unlike nurse cell nuclei, the growth of follicular epithelium nuclei is not accompanied with the proportional growth of the X chromosome. However, the shape of the X chromosome does not differ between the tissues. The dynamics of the X chromosome attachment regions location is tissue-specific and it is correlated with the process of nucleus growth in follicular epithelium and nurse cells. PMID:28158219

  6. 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

  7. Combined effect of seaweed (Sargassum wightii) and Bacillus thuringiensis var. israelensis on the coastal mosquito,Anopheles sundaicus, in Tamil Nadu, India

    Science.gov (United States)

    Studies were made of the extract of Sargassum wightii combined with Bacillus thuringiensis var. israelensis (Bti) for control of the malaria vector Anopheles sundaicus. Treatment of mosquito larvae with 0.001% S. wightii extract indicated median lethal concentrations (LC50) of 88, 73, 134, 156, and...

  8. Engineered resistance to Plasmodium falciparum development in transgenic Anopheles stephensi.

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    Alison T Isaacs

    2011-04-01

    Full Text Available Transposon-mediated transformation was used to produce Anopheles stephensi that express single-chain antibodies (scFvs designed to target the human malaria parasite, Plasmodium falciparum. The scFvs, m1C3, m4B7, and m2A10, are derived from mouse monoclonal antibodies that inhibit either ookinete invasion of the midgut or sporozoite invasion of salivary glands. The scFvs that target the parasite surface, m4B7 and m2A10, were fused to an Anopheles gambiae antimicrobial peptide, Cecropin A. Previously-characterized Anopheles cis-acting DNA regulatory elements were included in the transgenes to coordinate scFv production with parasite development. Gene amplification and immunoblot analyses showed promoter-specific increases in transgene expression in blood-fed females. Transgenic mosquito lines expressing each of the scFv genes had significantly lower infection levels than controls when challenged with P. falciparum.

  9. Successful human infection with P. falciparum using three aseptic Anopheles stephensi mosquitoes: a new model for controlled human malaria infection.

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    Matthew B Laurens

    Full Text Available Controlled human malaria infection (CHMI is a powerful method for assessing the efficacy of anti-malaria vaccines and drugs targeting pre-erythrocytic and erythrocytic stages of the parasite. CHMI has heretofore required the bites of 5 Plasmodium falciparum (Pf sporozoite (SPZ-infected mosquitoes to reliably induce Pf malaria. We reported that CHMI using the bites of 3 PfSPZ-infected mosquitoes reared aseptically in compliance with current good manufacturing practices (cGMP was successful in 6 participants. Here, we report results from a subsequent CHMI study using 3 PfSPZ-infected mosquitoes reared aseptically to validate the initial clinical trial. We also compare results of safety, tolerability, and transmission dynamics in participants undergoing CHMI using 3 PfSPZ-infected mosquitoes reared aseptically to published studies of CHMI using 5 mosquitoes. Nineteen adults aged 18-40 years were bitten by 3 Anopheles stephensi mosquitoes infected with the chloroquine-sensitive NF54 strain of Pf. All 19 participants developed malaria (100%; 12 of 19 (63% on Day 11. The mean pre-patent period was 258.3 hours (range 210.5-333.8. The geometric mean parasitemia at first diagnosis by microscopy was 9.5 parasites/µL (range 2-44. Quantitative polymerase chain reaction (qPCR detected parasites an average of 79.8 hours (range 43.8-116.7 before microscopy. The mosquitoes had a geometric mean of 37,894 PfSPZ/mosquito (range 3,500-152,200. Exposure to the bites of 3 aseptically-raised, PfSPZ-infected mosquitoes is a safe, effective procedure for CHMI in malaria-naïve adults. The aseptic model should be considered as a new standard for CHMI trials in non-endemic areas. Microscopy is the gold standard used for the diagnosis of Pf malaria after CHMI, but qPCR identifies parasites earlier. If qPCR continues to be shown to be highly specific, and can be made to be practical, rapid, and standardized, it should be considered as an alternative for diagnosis

  10. Population dynamics of sporogony for Plasmodium vivax parasites from western Thailand developing within three species of colonized Anopheles mosquitoes

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    Sattabongkot Jetsumon

    2006-08-01

    Full Text Available Abstract Background The population dynamics of Plasmodium sporogony within mosquitoes consists of an early phase where parasite abundance decreases during the transition from gametocyte to oocyst, an intermediate phase where parasite abundance remains static as oocysts, and a later phase where parasite abundance increases during the release of progeny sporozoites from oocysts. Sporogonic development is complete when sporozoites invade the mosquito salivary glands. The dynamics and efficiency of this developmental sequence were determined in laboratory strains of Anopheles dirus, Anopheles minimus and Anopheles sawadwongporni mosquitoes for Plasmodium vivax parasites circulating naturally in western Thailand. Methods Mosquitoes were fed blood from 20 symptomatic Thai adults via membrane feeders. Absolute densities were estimated for macrogametocytes, round stages (= female gametes/zygotes, ookinetes, oocysts, haemolymph sporozoites and salivary gland sporozoites. From these census data, five aspects of population dynamics were analysed; 1 changes in life-stage prevalence during early sporogony, 2 kinetics of life-stage formation, 3 efficiency of life-stage transitions, 4 density relationships between successive life-stages, and 5 parasite aggregation patterns. Results There was no difference among the three mosquito species tested in total losses incurred by P. vivax populations during early sporogony. Averaged across all infections, parasite populations incurred a 68-fold loss in abundance, with losses of ca. 19-fold, 2-fold and 2-fold at the first (= gametogenesis/fertilization, second (= round stage transformation, and third (= ookinete migration life-stage transitions, respectively. However, total losses varied widely among infections, ranging from 6-fold to over 2,000-fold loss. Losses during gametogenesis/fertilization accounted for most of this variability, indicating that gametocytes originating from some volunteers were more fertile than

  11. [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.

  12. Wolbachia surface protein induces innate immune responses in mosquito cells

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    Pinto Sofia B

    2012-01-01

    Full Text Available Abstract Background Wolbachia endosymbiotic bacteria are capable of inducing chronic upregulation of insect immune genes in some situations and this phenotype may influence the transmission of important insect-borne pathogens. However the molecules involved in these interactions have not been characterized. Results Here we show that recombinant Wolbachia Surface Protein (WSP stimulates increased transcription of immune genes in mosquito cells derived from the mosquito Anopheles gambiae, which is naturally uninfected with Wolbachia; at least two of the upregulated genes, TEP1 and APL1, are known to be important in Plasmodium killing in this species. When cells from Aedes albopictus, which is naturally Wolbachia-infected, were challenged with WSP lower levels of upregulation were observed than for the An. gambiae cells. Conclusions We have found that WSP is a strong immune elicitor in a naturally Wolbachia-uninfected mosquito species (Anopheles gambiae while a milder elicitor in a naturally-infected species (Aedes albopictus. Since the WSP of a mosquito non-native (nematode Wolbachia strain was used, these data suggest that there is a generalized tolerance to WSP in Ae. albopictus.

  13. Water management for controlling the breeding of Anopheles mosquitoes in rice irrigation schemes in Kenya.

    Science.gov (United States)

    Mutero, C M; Blank, H; Konradsen, F; van der Hoek, W

    2000-10-02

    An experiment to assess the impact of intermittent irrigation on Anopheles larval populations, rice yields and water use was conducted in the Mwea rice irrigation scheme in Kenya. Four water regimes including intermittent irrigation were tested in a complete randomized block experimental design. Intermittent irrigation was carried out on a weekly schedule, with flooded conditions from Saturday through Tuesday morning. Larval sampling at each plot was conducted every Monday and prior to draining of intermittently irrigated subplots on Tuesday. All the adult anopheline mosquitoes emerging from larvae collected in the experimental plots were identified as being An. arabiensis. By far the highest numbers of An. arabiensis 1st instar larvae were found in the intermittently irrigated subplots, indicating that the water regime provided the most attractive environment for egg laying. However, the ratio between the 4th and 1st instar larvae in the subplots was only 0.08, indicating very low survival rates. In contrast, the 4th/1st instar ratio for subplots with other water management regimes ranged between 0.27 and 0.68, suggesting a correspondingly higher survival than observed with intermittent irrigation. The total number of 4th instars was almost the same in the intermittently irrigated subplots and the irrigation system normally practised by the farmers. The failure to eliminate larval development up to the 4th instar in the former method was attributed to residual pools of water. Larval abundance fluctuated throughout the 12-week sampling period. The highest larval densities were recorded in the 3 weeks after transplanting the rice seedlings. Afterwards, larval numbers dropped dramatically as the height of rice plants increased. Rice yields at harvest did not show statistically significant differences among subplots with different water regimes. The average yield per hectare ranged from 4.8-5.3 metric tonnes. The average daily water percolation/seepage rate was 3.6 mm

  14. Mosquito larvicidal properties of volatile oil from salt marsh mangrove plant of Sesuvium portulacastrum against Anopheles stephensi and Aedes aegypti

    Institute of Scientific and Technical Information of China (English)

    Mohamed Yacoob Syed Ali; Venkatraman Anuradha; SyedAbudhair Sirajudeen; Prathasarathy Vijaya; Nagarajan Yogananth; Ramachandran Rajan; Peer Mohamed Kalitha Parveen

    2013-01-01

    Objective: To identify the larvicidal activity of the volatile oil from Sesuvium portulacastrum (S.portulacastrum ) against Anopheles stephensi and Aedes aegypti. Methods: Volatile oil extract of S. portulacastrum was prepared in a graded series of concentration. The test for the larvicidal effect of volatile oil against mosquitos larvae was conducted in accordance with the WHO standard method. Batches of 25 early 4th instar larvae of two mosquitoes were transferred to 250 mL enamel bowl containing 199 mL of distilled water and 1 mL of plant extracts. Each experiment was conducted with triplicate with concurrent a control group.Results:Volatile oil extract of S. portulacastrum showed toxicity against 4th instar larvae of Aedes aegypti and Anopheles stephensi with equivalent LC50 value [(63±7.8) µL/mL, LCL-UCL=55.2-64.0] and LC90 value [(94.2±3.9) µL/mL)] in maximum activity with minimum concentration (200 µL/mL) of volatile oil and followed by maximum activity of 250 µL concentration showed LC50 value=(68.0±8.2) µL/mL, LCL-UCL=66.26-69.2 and LC50 value of (55.2±2.8) µL/mL, LCL-UCL=53.7-56.9, LC90=(95.2±1.25) µL/mL and followed by 250 µL of oil extract against 4th instar larvae of Aedes aegypti respectively.Conclusions:It is inferred from the present study that, the extracts from salt marsh mangrove plan of S. portulacastum are identified as a potential source of safe and efficacious mosquito control agents for the management of vector borne diseases of malaria and dengue.

  15. 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.

  16. Heterosis Increases Fertility, Fecundity, and Survival of Laboratory-Produced F1 Hybrid Males of the Malaria Mosquito Anopheles coluzzii

    Science.gov (United States)

    Ekechukwu, Nkiru E.; Baeshen, Rowida; Traorè, Sékou F.; Coulibaly, Mamadou; Diabate, Abdoulaye; Catteruccia, Flaminia; Tripet, Frédéric

    2015-01-01

    The success of vector control strategies aiming to decrease disease transmission via the release of sterile or genetically-modified male mosquitoes critically depends on mating between laboratory-reared males and wild females. Unfortunately, mosquito colonization, laboratory rearing, and genetic manipulations can all negatively affect male competitiveness. Heterosis is commonly used to produce domestic animals with enhanced vigor and homogenous genetic background and could therefore potentially improve the mating performance of mass-reared male mosquitoes. Here, we produced enhanced hybrid males of the malaria mosquito Anopheles coluzzii by crossing two strains colonized >35 and 8 years ago. We compared the amount of sperm and mating plug proteins they transferred to females, as well as their insemination rate, reproductive success and longevity under various experimental conditions. Across experiments, widespread adaptations to laboratory mating were detected in the older strain. In large-group mating experiments, no overall hybrid advantage in insemination rates and the amount of sperm and accessory gland proteins transferred to females was detected. Despite higher sperm activity, hybrid males did not appear more fecund. However, individual-male mating and laboratory-swarm experiments revealed that hybrid males, while inseminating fewer females than older inbred males, were significantly more fertile, producing larger mating plugs and drastically increasing female fecundity. Heterotic males also showed increased longevity. These results validate the use of heterosis for creating hybrid males with improved fitness from long-established inbred laboratory strains. Therefore, this simple approach could facilitate disease control strategies based on male mosquito releases with important ultimate benefits to human health. PMID:26497140

  17. Susceptibility of Anopheles stephensi to Plasmodium gallinaceum: a trait of the mosquito, the parasite, and the environment.

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    Jen C C Hume

    Full Text Available BACKGROUND: Vector susceptibility to Plasmodium infection is treated primarily as a vector trait, although it is a composite trait expressing the joint occurrence of the parasite and the vector with genetic contributions of both. A comprehensive approach to assess the specific contribution of genetic and environmental variation on "vector susceptibility" is lacking. Here we developed and implemented a simple scheme to assess the specific contributions of the vector, the parasite, and the environment to "vector susceptibility." To the best of our knowledge this is the first study that employs such an approach. METHODOLOGY/PRINCIPAL FINDINGS: We conducted selection experiments on the vector (while holding the parasite "constant" and on the parasite (while holding the vector "constant" to estimate the genetic contributions of the mosquito and the parasite to the susceptibility of Anopheles stephensi to Plasmodium gallinaceum. We separately estimated the realized heritability of (i susceptibility to parasite infection by the mosquito vector and (ii parasite compatibility (transmissibility with the vector while controlling the other. The heritabilities of vector and the parasite were higher for the prevalence, i.e., fraction of infected mosquitoes, than the corresponding heritabilities of parasite load, i.e., the number of oocysts per mosquito. CONCLUSIONS: The vector's genetics (heritability comprised 67% of "vector susceptibility" measured by the prevalence of mosquitoes infected with P. gallinaceum oocysts, whereas the specific contribution of parasite genetics (heritability to this trait was only 5%. Our parasite source might possess minimal genetic diversity, which could explain its low heritability (and the high value of the vector. Notably, the environment contributed 28%. These estimates are relevant only to the particular system under study, but this experimental design could be useful for other parasite-host systems. The prospects and

  18. 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.

  19. Laboratory evaluation of traditional insect/mosquito repellent plants against Anopheles arabiensis, the predominant malaria vector in Ethiopia.

    Science.gov (United States)

    Karunamoorthi, Kaliyaperumal; Mulelam, Adane; Wassie, Fentahun

    2008-08-01

    Laboratory study was carried out to evaluate the repellent efficiency of most commonly known four traditional insect/mosquito repellent plants Wogert [vernacular name (local native language, Amharic); Silene macroserene], Kebercho [vernacular name (local native language, Amharic); Echinops sp.], Tinjut [vernacular name (local native language, Amharic); Ostostegia integrifolia], and Woira[vernacular name (local native language, Amharic); Olea europaea] against Anopheles arabiensis under the laboratory conditions. One hundred (4-5 days old) female A. arabiensis were introduced into the both 'control' and 'test' repellent chamber through the hole on top. Traditional charcoal stoves were used for direct burning. The experiment was conducted by applying the smoke into the repellent "test" mosquito cage by direct burning of 25 gm of dried plant materials (leaves and roots) until plant materials completely burned. The number of mosquitoes driving away from the "test" and "control" cage was recorded for every 5 min. In the present investigation, the results clearly revealed that the roots of S. macroserene has potent repellent efficiency (93.61%) and was the most effective. The leaves of Echinops sp. (92.47%), leaves of O. integrifolia (90.10%) and O. europaea (79.78%) were also effective. Roots of S. macroserene exhibited the highest repellent efficiency by direct burning. The present study identified these four traditional indigenous insect/mosquito repellent plant materials are very promising and can be used as safer alternative to modern synthetic chemical repellents against mosquito vectors of disease. Since people have been using these plants for some medicinal purposes, no side effects have been found.

  20. Infection of malaria (Anopheles gambiae s.s.) and filariasis (Culex quinquefasciatus) vectors with the entomopathogenic fungus Metarhizium anisopliae

    NARCIS (Netherlands)

    Scholte, E.J.; Njiru, B.N.; Smallegange, R.C.; Takken, W.; Knols, B.G.J.

    2003-01-01

    Background: Current intra-domiciliary vector control depends on the application of residual insecticides and/or repellents. Although biological control agents have been developed against aquatic mosquito stages, none are available for adults. Following successful use of an entomopathogenic fungus ag

  1. Beer Consumption Increases Human Attractiveness to Malaria Mosquitoes

    Science.gov (United States)

    Lefèvre, Thierry; Gouagna, Louis-Clément; Dabiré, Kounbobr Roch; Elguero, Eric; Fontenille, Didier; Renaud, François; Costantini, Carlo; Thomas, Frédéric

    2010-01-01

    Background Malaria and alcohol consumption both represent major public health problems. Alcohol consumption is rising in developing countries and, as efforts to manage malaria are expanded, understanding the links between malaria and alcohol consumption becomes crucial. Our aim was to ascertain the effect of beer consumption on human attractiveness to malaria mosquitoes in semi field conditions in Burkina Faso. Methodology/Principal Findings We used a Y tube-olfactometer designed to take advantage of the whole body odour (breath and skin emanations) as a stimulus to gauge human attractiveness to Anopheles gambiae (the primary African malaria vector) before and after volunteers consumed either beer (n = 25 volunteers and a total of 2500 mosquitoes tested) or water (n = 18 volunteers and a total of 1800 mosquitoes). Water consumption had no effect on human attractiveness to An. gambiae mosquitoes, but beer consumption increased volunteer attractiveness. Body odours of volunteers who consumed beer increased mosquito activation (proportion of mosquitoes engaging in take-off and up-wind flight) and orientation (proportion of mosquitoes flying towards volunteers' odours). The level of exhaled carbon dioxide and body temperature had no effect on human attractiveness to mosquitoes. Despite individual volunteer variation, beer consumption consistently increased attractiveness to mosquitoes. Conclusions/Significance These results suggest that beer consumption is a risk factor for malaria and needs to be integrated into public health policies for the design of control measures. PMID:20209056

  2. Beer consumption increases human attractiveness to malaria mosquitoes.

    Directory of Open Access Journals (Sweden)

    Thierry Lefèvre

    Full Text Available BACKGROUND: Malaria and alcohol consumption both represent major public health problems. Alcohol consumption is rising in developing countries and, as efforts to manage malaria are expanded, understanding the links between malaria and alcohol consumption becomes crucial. Our aim was to ascertain the effect of beer consumption on human attractiveness to malaria mosquitoes in semi field conditions in Burkina Faso. METHODOLOGY/PRINCIPAL FINDINGS: We used a Y tube-olfactometer designed to take advantage of the whole body odour (breath and skin emanations as a stimulus to gauge human attractiveness to Anopheles gambiae (the primary African malaria vector before and after volunteers consumed either beer (n = 25 volunteers and a total of 2500 mosquitoes tested or water (n = 18 volunteers and a total of 1800 mosquitoes. Water consumption had no effect on human attractiveness to An. gambiae mosquitoes, but beer consumption increased volunteer attractiveness. Body odours of volunteers who consumed beer increased mosquito activation (proportion of mosquitoes engaging in take-off and up-wind flight and orientation (proportion of mosquitoes flying towards volunteers' odours. The level of exhaled carbon dioxide and body temperature had no effect on human attractiveness to mosquitoes. Despite individual volunteer variation, beer consumption consistently increased attractiveness to mosquitoes. CONCLUSIONS/SIGNIFICANCE: These results suggest that beer consumption is a risk factor for malaria and needs to be integrated into public health policies for the design of control measures.

  3. Mosquito repellent potential ofPithecellobium dulce leaf and seed against malaria vectorAnopheles stephensi (Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    Mohan Rajeswary; Marimuthu Govindarajan

    2016-01-01

    Objective:To determine the repellent properties of hexane, benzene, ethyl acetate, chloroform and methanol extract ofPithecellobium dulce (P. dulce) leaf and seed against Anopheles stephensi(An. stephensi). Methods:Repellent activity assay was carried out in a net cage (45 cmí30 cmí25 cm) containing 100 blood starved female mosquitoes ofAn. stephensi. This assay was carried out in the laboratory conditions according to theWHO 2009 protocol. Plant crude extracts ofP. 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 ofP. dulceprovided over 180 min and 150 min protection, respectively. Conclusions: Crude extracts ofP. dulceexhibit the potential for controlling malaria vector mosquitoAn. stephensi.

  4. Mosquito larvicidal and biting deterrency activity of bud of Polianthes tuberosa plants extract against Anopheles stephensi and Culex quinquefasciatus.

    Science.gov (United States)

    Anjali, Rawani; Atanu, Banerjee; Goutam, Chandra

    2012-06-01

    Mosquito control by phytochemicals is an alternative method to synthetic insecticides, as it is biodegradable and non resistant to vector mosquito. Polianthes tuberosa is a perennial plant distributed in many parts of India. The present study was undertaken to scientifically evaluate the larvicide and biting deterrency activity of bud of Polianthes tuberosa against Culex quinquefasciatus and Anopheles stephensi. Crude and solvent extract [ethyl acetate, chloroform: methanol (1:1, v/v), acetone] of fresh, mature, bud of P. tuberosa was tested against (ex. quinquefasciatus and An. stephensi. The repellent activity tested by chloroform: methanol (1:1, v/v) solvent extract against both mosquito species. The appropriate lethal concentrations at 24h for chloroform: methanol (1:1, v/v) extract was also studied on non target organisms such as Toxorhynchites larvae, Diplonychus annulatum and Chironomus circumdatus. In a 72 hour bioassay experiment, 0.5% crude extract showed the highest mortality and chloroform: methanol (1:1, v/v) solvent extract showed the highest mortality, the maximum (p larvicide agent. There is no changes in the activity non-target organism so, it is safe to use.

  5. The molecular evolution of four anti-malarial immune genes in the Anopheles gambiae species complex - art. no. 79

    OpenAIRE

    Parmakelis, A.; Slotman, M. A.; Marshall, J. C.; Awono Ambene, P. H.; Antonio Nkondjio, C.; Simard, Frédéric; Caccone, A; Powell, J. R.

    2008-01-01

    Background: If the insect innate immune system is to be used as a potential blocking step in transmission of malaria, then it will require targeting one or a few genes with highest relevance and ease of manipulation. The problem is to identify and manipulate those of most importance to malaria infection without the risk of decreasing the mosquito's ability to stave off infections by microbes in general. Molecular evolution methodologies and concepts can help identify such genes. Within the se...

  6. Mosquito mass rearing technology: a cold-water vortex device for continuous unattended separation of Anopheles arabiensis pupae from larvae.

    Science.gov (United States)

    Balestrino, Fabrizio; Gilles, Jérémie R L; Soliban, Sharon M; Nirschl, Anton; Benedict, Quentin E; Benedict, Mark Q

    2011-09-01

    In mass rearing of anopheline mosquitoes, pupae are usually separated from larvae on a daily basis to prevent unwanted adult emergence from trays. Depending on the device and species, 2 physical characteristics have most often been used for separation: buoyant density and size. In this report, we describe a system for continuous separation of Anopheles arabiensis larvae from pupae based on the natural difference in buoyant density and behavior between the 2 stages. We determined that temperatures 4-15 degrees C caused neither mortality nor reduction in likelihood of pupation or emergence. Separation improved as temperatures decreased down to 4 degrees C. We devised and demonstrated a 15 degrees C water vortex separator that we anticipate can process approximately 1 million larvae and pupae per hour with a < 0.3% pupal contamination rate and which operates unattended.

  7. 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.

  8. Mosquito larvicidal activity of some common spices and vegetable waste onCulex quinquefasciatus andAnopheles stephensi

    Institute of Scientific and Technical Information of China (English)

    Someshwar Singha; Goutam Chandra

    2011-01-01

    Objective:To investigate the larvicidal activities of crude and chloroform: methanol (1:1 v/v) extracts of some common spices (Cuminum cyminum, Allium sativum, Zingiber offinale, Curcuma longa) and vegetable waste (Solanum tuberosum germinated tuber) againstAnopheles stephensiandCulex quinquefasciatus mosquito larvae.Methods:Larval mortality of above mosquito species were observed after 24, 48 and72 h of exposure to five concentrations of aqueous extract (0.1%, 0.2%, 0.3%, 0.4% and0.5%) and four concentrations (25, 50, 75 ppm) of chloroform: methanol (1:1 v/v) extract. The lethal concentration of individual spices or vegetable waste was determined by log-probit analysis (at95%confidence level) and effect of crude and chloroform: methanol (1:1 v/v) extracts were recorded on non target organisms.Results:Relative mortality rate of both larval mosquito species were recorded in the following sequences:Cuminum cyminum>Allium sativum>Zingiber offinale, Curcuma longa > Solanum tuberosum germinated tuber for crude extract, and efficacy of chloroform: methanol (1:1 v/v) extract were as follows:Curcuma longa > Zingiber offinale > Solanum tuberosum germinated tuber >Cuminum cyminum>Allium sativum.Conclusions: Crude and chloroform: methanol (1:1 v/v) extract of Cuminum cyminum, Allium sativum, Zingiber offinale, Curcuma longa andSolanum tuberosum germinated tuber can be recommended effectively in mosquito control programmes at very low concentrations. No mortality and other abnormalities were noticed on non target organisms and further studies are needed to investigate the chemical structure of active principal which are responsible for larvicidal activity.

  9. A 15N stable isotope semen label to detect mating in the malaria mosquito Anopheles arabiensis Patton

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    Gludovacz Doris

    2008-07-01

    Full Text Available Abstract In previous studies it was determined that the stable isotope 13-carbon can be used as a semen label to detect mating events in the malaria mosquito Anopheles arabiensis. In this paper we describe the use of an additional stable isotope, 15-nitrogen (15N, for that same purpose. Both stable isotopes can be analysed simultaneously in a mass spectrometer, offering the possibility to detect both labels in one sample in order to study complex and difficult-to-detect mating events, such as multiple mating. 15N-glycine was added to larval rearing water and the target enrichment was 5 atom% 15N. Males from these trays were mated with unlabelled virgin females, and spiked spermathecae were analysed for isotopic composition after mating using mass spectrometry. Results showed that spermathecae positive for semen could be distinguished from uninseminated or control samples using the raw δ15N‰ values. The label persisted in spermathecae for up to 5 days after insemination, and males aged 10 days transferred similar amounts of label as males aged 4 days. There were no negative effects of the label on larval survival and male longevity. Enrichment of teneral mosquitoes after emergence was 4.85 ± 0.10 atom% 15N. A threshold value defined as 3 standard deviations above the mean of virgin (i.e. uninseminated spermathecae samples was successful in classifying a large proportion of samples correctly (i.e. on average 95%. We conclude that alongside 13C, 15N can be used to detect mating in Anopheles and the suitability of both labels is briefly discussed.

  10. 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.

  11. A comprehensive transcriptomic view of renal function in the malaria vector, Anopheles gambiae

    DEFF Research Database (Denmark)

    Overend, Gayle; Cabrero, Pablo; Halberg, Kenneth Agerlin

    2015-01-01

    Renal function is essential to maintain homeostasis. This is particularly significant for insects that undergo complete metamorphosis; larval mosquitoes must survive a freshwater habitat whereas adults are terrestrial, and mature females must maintain ion and fluid homeostasis after blood feeding......, and generated RNAi knockdown flies. Three of these genes proved essential, showing conservation of critical functions across 150 million years of phylogenetic separation. This extensive data-set is available as an online resource, MozTubules.org, and could potentially be mined for novel insecticide targets...

  12. Mosquito larvicidal properties ofOrthosiphon thymiflorus(Roth) Sleesen. (Family:Labiatae) against mosquito vectors,Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti (Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    K Kovendan; K Murugan; S Vincent; Donald R Barnard

    2012-01-01

    ABSTRACT Objective:To determine the mosquito larvicidal activities of hexane, chloroform, ethyl acetate, acetone and methanol leaf extract ofOrthosiphon thymiflorus (O. thymiflorus) againstAnopheles stephensi (An. stephensi), Culex quinquefasciatus (Cx. quinquefasciatus) andAedes aegypti (Ae. aegypti).Methods: The larvicidal activity was assayed against three mosquito species at various concentrations ranging from (50-450 ppm) under the laboratory conditions. TheLC50and LC90value of theO. thymiflorus leaf extract was determined by Probit analysis.Results: The LC50values of hexane, chloroform, ethyl acetate, acetone and methanol extract ofO. thymiflorus third instar larvae ofAn. stephensiwereLC50= 201.39, 178.76, 158.06, 139.22 and118.74 ppm;Cx. quinquefasciatus were LC50=228.13, 209.72, 183.35, 163.55 and149.96 ppm andAe. aegyptiwere LC50=215.65, 197.91, 175.05, 154.80 and137.26 ppm, respectively. Maximum larvicidal activity was observed in the methanolic extract followed by acetone, ethyl acetate chloroform and hexane extract. The larval mortality was observed after24h exposure. No mortality was observed in control.Conclusions:The present results suggest that the effective plant crude extracts have potential to be used as an ideal eco-friendly approach for the control of mosquito vectors. This study provides the first report on the larvicidal activity of this plant crude solvent extract of againstAn. stephensi, Cx. quinquefasciatus andAe. aegyptimosquitoes.

  13. 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.

  14. Ammonium sulphate fertiliser increases larval populations of Anopheles arabiensis and culicine mosquitoes in rice fields

    DEFF Research Database (Denmark)

    Mutero, C M; Ng'ang'a, P N; Wekoyela, P;

    2004-01-01

    Field experiments were conducted in central Kenya, to study the effect of ammonium sulphate fertiliser ((NH(4))(2)SO(4)) on mosquito larval populations in rice fields. The experiments used a complete randomised block design having four blocks with two experimental ponds per block...... in rice fields, thereby making them visually more attractive for egg-laying by An. arabiensis and culicine mosquitoes....

  15. Gustatory receptor neuron responds to chemically diverse insect repellents in the common malaria mosquito Anopheles quadrimaculatus

    Science.gov (United States)

    Female mosquitoes feed on blood from animal hosts to obtain nutritional resources used for egg production. These contacts facilitate the spread of harmful human diseases. Chemical repellents are used to disrupt mosquito host seeking and blood feeding behaviors; however, little is known about the g...

  16. Anopheles (Nyssorhynchus) Pictipennis: A New Mosquito Record from the Atacama Region of Northern Chile

    Science.gov (United States)

    2008-01-01

    Santiago, this species rarely feeds on humans, but it is highly attracted to domestic animals (Linthicum 1988). In this paper, we report An. (N...Mosq Control Assoc 12:619-626. Neghme AR. 1943. Contribucion a la biologia del Anopheles pictipenllis (Philippi, 1865). I. Communica- cion

  17. Mosquito larvicidal and ovicidal activity of puffer fish extracts against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti (Diptera: Culicidae).

    Science.gov (United States)

    Samidurai, Kaliyaperumal; Mathew, Nisha

    2013-03-01

    The extracts of liver (LE), ovary (OE), skin (SE) and muscle (ME) tissues of four species of puffer fishes viz., Arothron hispidus, Lagocephalus inermis, Lagocephalus scleratus and Chelonodon patoca were evaluated against larvae and eggs of three mosquito vectors, Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. The LC50 values were 1194.26, 1382.73 (LE); 1421.42, 1982.73 (OE); 7116.86, 15038.98 (ME) and 10817.8 ppm (SE) for An. stephensi and Cx. quinquefasciatus respectively for A. hispidus. In the case of L. inermis, the LC50 values were 1163.83, 1556.1 and 2426.38 (LE); 1653.53, 2734.74 (OE); 6067.47 (ME) and 10283.04 ppm (SE) for An. stephensi, Cx. quinquefasciatus and Ae. aegypti respectively. The LC50 values were 1509.98, 1608.69 (LE) and 1414.9, 2278.69 ppm (OE) for An. stephensi and Cx. quinquefasciatus respectively for the extracts of L. scleratus. In the case C. patoca extracts the LC50 values were 1182.29, 1543.00, 2441.03 (LE) and 1076.13, 2582.11 ppm (OE) for An. stephensi, Cx. quinquefasciatus and Ae. aegypti respectively. OE and LE of all puffer fishes exhibited zero percent egg hatchability from 600 to 1000 ppm against eggs of An. stephensi and Cx. quinquefasciatus. This study shows that puffer toxins are effective in killing the larvae and eggs of mosquitoes.

  18. Recombinant Pvs48/45 antigen expressed in E. coli generates antibodies that block malaria transmission in Anopheles albimanus mosquitoes.

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    Myriam Arévalo-Herrera

    Full Text Available Transmission of malaria parasites from humans to Anopheles mosquitoes can be inhibited by specific antibodies elicited during malaria infection, which target surface Plasmodium gametocyte/gamete proteins. Some of these proteins may have potential for vaccine development. Pvs48/45 is a P. vivax gametocyte surface antigen orthologous to Pfs48/45, which may play a role during parasite fertilization and thus has potential for transmission blocking (TB activity. Here we describe the expression of a recombinant Pvs48/45 protein expressed in Escherichia coli as a ∼60kDa construct which we tested for antigenicity using human sera and for its immunogenicity and transmission blocking activity of specific anti-mouse and anti-monkey Pvs48/45 antibodies. The protein reacted with sera of individuals from malaria-endemic areas and in addition induced specific IgG antibody responses in BALB/c mice and Aotus l. griseimembra monkeys. Sera from both immunized animal species recognized native P. vivax protein in Western blot (WB and immunofluorescence assays. Moreover, sera from immunized mice and monkeys produced significant inhibition of parasite transmission to An. Albimanus mosquitoes as shown by membrane feeding assays. Results indicate the presence of reactive epitopes in the Pvs48/45 recombinant product that induce antibodies with TB activity. Further testing of this protein is ongoing to determine its vaccine potential.

  19. 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.

  20. Efficacy of leaves extract of Calotropis procera Ait. (Asclepiadaceae) in controlling Anopheles arabiensis and Culex quinquefasciatus mosquitoes.

    Science.gov (United States)

    Elimam, Abdalla M; Elmalik, Khitma H; Ali, Faysal S

    2009-10-01

    The present study aimed to investigate, the larvicidal, adult emergence inhibition and oviposition deterrent activity of aqueous leaves extract of Calotropis procera against Anopheles arabiensis and Culex quinquefasciatus as natural mosquito larvicide. The larvicidal activity was monitored against 2nd, 3rd and 4th instar larvae of each mosquito species 24 h post-treatment. Adult emergence inhibition activity was tested by exposing 3rd instar larvae of each mosquito species to different concentrations of extracts (200, 400, 600, 800 and 1000 ppm for An. arabiensis and 100, 200, 300, 400, 500 and 600 ppm for Cx. quinquefasciatus). Probit analysis was used to analyze data from bioassay experiments. The oviposition deterrent activity was tested by using three different concentrations of extracts (1000, 500 and 200 for An. arabiensis, and 1000, 500 and 100 for Cx. quinquefasciatus) that caused high, moderate and low larval mortality in the larvicidal experiment against 3rd instar larvae. It was found that, LC50-LC90 values calculated were 273.53-783.43, 366.44-1018.59 and 454.99-1224.62 ppm for 2nd, 3rd and 4th larval instars, respectively, of An. arabiensis and 187.93-433.51, 218.27-538.27 and 264.85-769.13 ppm for 2nd, 3rd and 4th larval instars, respectively, of Cx. quinquefasciatus. Fifty percent of adult emergence inhibition (EI50) was shown at 277.90 and 183.65 ppm for An. arabiensis and Cx. quinquefasciatus, respectively. The pupal stage was not affected till a concentration of 5000 ppm. The extract showed oviposition deterrence and effective repellence against both mosquito species at different concentrations, with the observation on that maximal eggs were laid in low concentration of extract. These results suggest that the leaves extract of C. procera possess remarkable larvicidal, adult emergence inhibitor, repellent and oviposition deterrent effect against both An. arabiensis and Cx. quinquefasciatus, and might be used as natural biocides for mosquito

  1. Mosquito Larvicidal Potential of Gossypium hirsutum (Bt cotton Leaves Extracts against Aedes aegypti and Anopheles stephensi larvae.

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    Chandrashekhar D Patil

    2014-06-01

    Full Text Available We aimed to extract the ingredients from leaves of Gossypium hirsutum (Bt cotton using different solvents and evaluate for potential use to control different larval stages of mosquito species, Aedes aegypti and Anopheles stephensi.Qualitative and quantitative estimation of ingredients from Go. hirsutum (Bt plant extract was carried out and their inhibitory action against mosquito larvae was determined using mosquito larvicidal assay.LC50 values of water, ethanol, ethyl acetate and hexane extracts for Ae. aegypti were 211.73±21.49, 241.64±19.92, 358.07±32.43, 401.03±36.19 and 232.56±26.00, 298.54±21.78, 366.50±30.59, 387.19±31.82 for 4(th instar of An. stephensi, respectively. The water extract displayed lowest LC50 value followed by ethanol, ethyl acetate and hexane. Owing to the comparatively better activity of water extract, its efficacy was further evaluated for mosquito larvicidal activity, which exhibited LC50 values of 133.95±12.79, 167.65±11.34 against 2(nd and 3(rd instars of Ae. aegypti and 145.48±11.76, 188.10±12.92 against 2(nd and 3(rd instars of An. stephensi, respectively. Crude protein from the water extract was precipitated using acetone and tested against 2(nd, 3(rd and 4(th instars of Ae. aegypti and An. stephensi. It revealed further decrease in LC50 values as 105.72±25.84, 138.23±23.18, 126.19±25.65, 134.04±04 and 137.88±17.59, 154.25±16.98 for 2(nd, 3(rd and 4(th instars of Ae. aegypti and An. stephensi, respectively.Leaves extracts of Go. hirsutum (Bt is potential mosquito larvicide and can be used as a potent alternative to chemical insecticides in integrated pest management.

  2. 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

  3. A Genome-Scale Investigation of Incongruence in Culicidae Mosquitoes.

    Science.gov (United States)

    Wang, Yuyu; Zhou, Xiaofan; Yang, Ding; Rokas, Antonis

    2015-12-01

    Comparison of individual gene trees in several recent phylogenomic studies from diverse lineages has revealed a surprising amount of topological conflict or incongruence, but we still know relatively little about its distribution across the tree of life. To further our understanding of incongruence, the factors that contribute to it and how it can be ameliorated, we examined its distribution in a clade of 20 Culicidae mosquito species through the reconstruction and analysis of the phylogenetic histories of 2,007 groups of orthologous genes. Levels of incongruence were generally low, the three exceptions being the internodes concerned with the branching of Anopheles christyi, with the branching of the subgenus Anopheles as well as the already reported incongruence within the Anopheles gambiae species complex. Two of these incongruence events (A. gambiae species complex and A. christyi) are likely due to biological factors, whereas the third (subgenus Anopheles) is likely due to analytical factors. Similar to previous studies, the use of genes or internodes with high bootstrap support or internode certainty values, both of which were positively correlated with gene alignment length, substantially reduced the observed incongruence. However, the clade support values of the internodes concerned with the branching of the subgenus Anopheles as well as within the A. gambiae species complex remained very low. Based on these results, we infer that the prevalence of incongruence in Culicidae mosquitoes is generally low, that it likely stems from both analytical and biological factors, and that it can be ameliorated through the selection of genes with strong phylogenetic signal. More generally, selection of genes with strong phylogenetic signal may be a general empirical solution for reducing incongruence and increasing the robustness of inference in phylogenomic studies.

  4. Efficacy of PermaNet® 2.0 and PermaNet® 3.0 against insecticide-resistant Anopheles gambiae in experimental huts in Côte d'Ivoire

    Directory of Open Access Journals (Sweden)

    Koffi Alphonsine A

    2011-06-01

    Full Text Available Abstract Background Pyrethroid resistance in vectors could limit the efficacy of long-lasting insecticidal nets (LLINs because all LLINs are currently treated with pyrethroids. The goal of this study was to evaluate the efficacy and wash resistance of PermaNet® 3.0 compared to PermaNet® 2.0 in an area of high pyrethroid in Côte d'Ivoire. PermaNet® 3.0 is impregnated with deltamethrin at 85 mg/m2 on the sides of the net and with deltamethrin and piperonyl butoxide on the roof. PermaNet® 2.0 is impregnated with deltamethrin at 55 mg/m2 across the entire net. Methods The study was conducted in the station of Yaokoffikro, in central Côte d'Ivoire. The efficacy of intact unwashed and washed LLINs was compared over a 12-week period with a conventionally-treated net (CTN washed to just before exhaustion. WHO cone bioassays were performed on sub-sections of the nets, using wild-resistant An. gambiae and Kisumu strains. Mosquitoes were collected five days per week and were identified to genus and species level and classified as dead or alive, then unfed or blood-fed. Results Mortality rates of over 80% from cone bioassays with wild-caught pyrethroid-resistant An. gambiae s.s were recorded only with unwashed PermaNet® 3.0. Over 12 weeks, a total of 7,291 mosquitoes were collected. There were significantly more An. gambiae s.s. and Culex spp. caught in control huts than with other treatments (P An. gambiae s.s and Culex spp, were lower for the control than for other treatments (P 0.05 except for unwashed PermaNet® 3.0 (P Conclusions This study showed that unwashed PermaNet® 3.0 caused significantly higher mortality against pyrethroid resistant An. gambiae s.s and Culex spp than PermaNet® 2.0 and the CTN. The increased efficacy with unwashed PermaNet® 3.0 over PermaNet® 2.0 and the CTN was also demonstrated by higher KD and mortality rates (KD > 95% and mortality rate > 80% in cone bioassays performed with wild pyrethroid-resistant An. gambiae s

  5. Larvicidal property of green synthesized silver nanoparticles against vector mosquitoes (Anopheles stephensi and Aedes aegypti)

    OpenAIRE

    Rajamani Bhuvaneswari; Raju John Xavier; Manickam Arumugam

    2016-01-01

    Mosquito vectors spread severe human diseases which lead to millions of deaths every year. Vector management is ultimately aimed to develop the health of every individual’s life by reducing the mosquito diversity. Control of vectors in growing counties is an important issue with various aspects. The advancement of green nanotechnology will attribute the solution for vector controlling policy. To identify the larvicidal property of silver nanoparticles (AgNPs) using Belosynapsis kewensis (B. k...

  6. Chemical composition and laboratory investigation of Melissa officinalis essential oil against human malarial vector mosquito, Anopheles stephensi L. (Diptera: Culicidae

    Directory of Open Access Journals (Sweden)

    Mathalaimuthu Baranitharan

    2016-11-01

    Full Text Available Objective: To decide the larvicides, ovicidal, pupicidal and repellent activity of Melissa officinalis (M. officinalis chemical compositions against important mosquito Anopheles stephensi (An. stephensi (Diptera: Culicidae. Methods: A chemical constituent of 24 compounds was identified in the oils of M. officinalis compounds representing to 98.73%. A total of 25 3rd instar larvae of An. stephensi were showed to a variety of concentrations (30–300 mg/L in laboratory by means of utilizing the standard procedure portrayed by World Health Organization (2005. The larvae were exposed for 24 h and mortalities were subjected to probit analysis. The ovicidal activity was strong-minded against An. stephensi mosquito to a variety of concentrations ranging from 15– 90 mg/L under the laboratory circumstances. The repellent activity of M. officinalis chemical compositions tested at concentrations of 0.75 and 1.50 mg/cm2 was evaluated in a net cage (45 cm × 45 cm × 40 cm including 100 blood starving female mosquitoes of An. stephensi using the methods of World Health Organization (1996. Results: The LC50 and LC90 values of citronellal compound against An. stephensi larvae were 85.44 and 159.73 mg/L, respectively. Mean percent hatchability of the ovicidal action was observed 48 h post-treatment. Similarly, the citronellal compound and other compositions were found to be mainly effective against eggs of An. stephensi. Citronellal compound exerted 45, 60, 75 and 90 mg/L against An. stephensi, respectively. The repellent activity of citronellal compound was contained to be mainly effective and the maximum action was observed at 0.75 and 1.50 mg/cm2 concentrations giving 100% protection up to 210 min against An. stephensi. Conclusions: This current study was undertaken to evaluate the larvicidal, ovicidal, repellent potential of compounds from the M. officinalis essential oil against An. stephensi. This is initial statement on the mosquito larvicidal

  7. Radical remodeling of the Y chromosome in a recent radiation of malaria mosquitoes.

    Science.gov (United States)

    Hall, Andrew Brantley; Papathanos, Philippos-Aris; Sharma, Atashi; Cheng, Changde; Akbari, Omar S; Assour, Lauren; Bergman, Nicholas H; Cagnetti, Alessia; Crisanti, Andrea; Dottorini, Tania; Fiorentini, Elisa; Galizi, Roberto; Hnath, Jonathan; Jiang, Xiaofang; Koren, Sergey; Nolan, Tony; Radune, Diane; Sharakhova, Maria V; Steele, Aaron; Timoshevskiy, Vladimir A; Windbichler, Nikolai; Zhang, Simo; Hahn, Matthew W; Phillippy, Adam M; Emrich, Scott J; Sharakhov, Igor V; Tu, Zhijian Jake; Besansky, Nora J

    2016-04-12

    Y chromosomes control essential male functions in many species, including sex determination and fertility. However, because of obstacles posed by repeat-rich heterochromatin, knowledge of Y chromosome sequences is limited to a handful of model organisms, constraining our understanding of Y biology across the tree of life. Here, we leverage long single-molecule sequencing to determine the content and structure of the nonrecombining Y chromosome of the primary African malaria mosquito, Anopheles gambiae We find that the An. gambiae Y consists almost entirely of a few massively amplified, tandemly arrayed repeats, some of which can recombine with similar repeats on the X chromosome. Sex-specific genome resequencing in a recent species radiation, the An. gambiae complex, revealed rapid sequence turnover within An. gambiae and among species. Exploiting 52 sex-specific An. gambiae RNA-Seq datasets representing all developmental stages, we identified a small repertoire of Y-linked genes that lack X gametologs and are not Y-linked in any other species except An. gambiae, with the notable exception of YG2, a candidate male-determining gene. YG2 is the only gene conserved and exclusive to the Y in all species examined, yet sequence similarity to YG2 is not detectable in the genome of a more distant mosquito relative, suggesting rapid evolution of Y chromosome genes in this highly dynamic genus of malaria vectors. The extensive characterization of the An. gambiae Y provides a long-awaited foundation for studying male mosquito biology, and will inform novel mosquito control strategies based on the manipulation of Y chromosomes.

  8. Laminin and the malaria parasite's journey through the mosquito midgut.

    Science.gov (United States)

    Arrighi, Romanico B G; Lycett, Gareth; Mahairaki, Vassiliki; Siden-Kiamos, Inga; Louis, Christos

    2005-07-01

    During the invasion of the mosquito midgut epithelium, Plasmodium ookinetes come to rest on the basal lamina, where they transform into the sporozoite-producing oocysts. Laminin, one of the basal lamina's major components, has previously been shown to bind several surface proteins of Plasmodium ookinetes. Here, using the recently developed RNAi technique in mosquitoes, we used a specific dsRNA construct targeted against the LANB2 gene (laminin gamma1) of Anopheles gambiae to reduce its mRNA levels, leading to a substantial reduction in the number of successfully developed oocysts in the mosquito midgut. Moreover, this molecular relationship is corroborated by the intimate association of developing P. berghei parasites and laminin in the gut, as observed using confocal microscopy. Our data support the notion of laminin playing a functional role in the development of the malaria parasite within the mosquito midgut.

  9. Infection of the malaria mosquito Anopheles gambiae with the entomopathogenic fungus Metarhizium anisopliae reduces blood feeding and fecundity

    NARCIS (Netherlands)

    Scholte, E.J.; Knols, B.G.J.; Takken, W.

    2006-01-01

    The entomopathogenic fungus Metarhizium anisopliae is being considered as a biocontrol agent against adult African malaria vectors. In addition to causing significant mortality, this pathogen is known to cause reductions in feeding and fecundity in a range of insects. In the present study we investi

  10. Infection of Anopheles gambiae mosquitoes with entomopathogenic fungi: effect of host age and blood-feeding status

    NARCIS (Netherlands)

    L.L. Mnyone; M.J. Kirby; M.W. Mpingwa; D.W. Lwetoijera; B.G.J. Knols; W. Takken; C.J.M. Koenraadt; T.L. Russell

    2011-01-01

    Physiological characteristics of insects can influence their susceptibility to fungal infection of which age and nutritional status are among the most important. An understanding of host-pathogen interaction with respect to these physiological characteristics of the host is essential if we are to de

  11. A Multi-detection Assay for Malaria Transmitting Mosquitoes

    Science.gov (United States)

    Lee, Yoosook; Weakley, Allison M.; Nieman, Catelyn C.; Malvick, Julia; Lanzaro, Gregory C.

    2015-01-01

    The Anopheles gambiae species complex includes the major malaria transmitting mosquitoes in Africa. Because these species are of such medical importance, several traits are typically characterized using molecular assays to aid in epidemiological studies. These traits include species identification, insecticide resistance, parasite infection status, and host preference. Since populations of the Anopheles gambiae complex are morphologically indistinguishable, a polymerase chain reaction (PCR) is traditionally used to identify species. Once the species is known, several downstream assays are routinely performed to elucidate further characteristics. For instance, mutations known as KDR in a para gene confer resistance against DDT and pyrethroid insecticides. Additionally, enzyme-linked immunosorbent assays (ELISAs) or Plasmodium parasite DNA detection PCR assays are used to detect parasites present in mosquito tissues. Lastly, a combination of PCR and restriction enzyme digests can be used to elucidate host preference (e.g., human vs. animal blood) by screening the mosquito bloodmeal for host-specific DNA. We have developed a multi-detection assay (MDA) that combines all of the aforementioned assays into a single multiplex reaction genotyping 33SNPs for 96 or 384 samples at a time. Because the MDA includes multiple markers for species, Plasmodium detection, and host blood identification, the likelihood of generating false positives or negatives is greatly reduced from previous assays that include only one marker per trait. This robust and simple assay can detect these key mosquito traits cost-effectively and in a fraction of the time of existing assays. PMID:25867057

  12. 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

  13. Comparative mosquito repellency of essential oils against Aedes aegypti (Linn.), Anopheles dirus (Peyton and Harrison) and Culex quinquefasciatus (Say)

    Institute of Scientific and Technical Information of China (English)

    Siriporn Phasomkusolsil; Mayura Soonwera

    2011-01-01

    Objective: To assess the repellency to female Aedes aegypti (Ae. aegypti), Anopheles dirus (An. dirus) and Culex quinquefasciatus (Cx. quinquefasciatus) of seven essential oils using two treatment methods. Methods: Topical applications of three dose concentrations (0.02, 0.10 and 0.21 mg/cm2) were made on the forearms of volunteers. Dose-response study and protection time study were employed in the experiment. Results: In the dose-response test, Cymbopogon citratus (C. citratus), Cymbopogon nardus (C. nardus), Syzygium aromaticum (S. aromaticum) and Ocimum basilicum (O. basilicum) exhibited a high repellency against Ae. aegypti with ED50 at < 0.045 mg/cm2, whereas C. citratus, C. nardus and S. aromaticum showed repellency against An. dirus with ED50 at <0.068 mg/cm2. Furthermore, the essential oils of C. citratus, C. nardus, S. aromaticum, O.basilicum and Cananga odorata gave strong effective dose (ED 50) values at <0.003 mg/cm2 when tested against Cx. quinquefasciatus. For testing by arm in cage method, at 0.21 mg/cm2, protection time of C. citratus gave the longest lasting period against three mosquito species, 72 min for Ae. aegypti, 132 min for An. dirus and 84 min for Cx. quinquefasciatus. In addition, the two essential oils exhibited moderate repellency against Ae. aegypti, An. dirus and Cx. quinquefasciatus, at 60, 90 and 78 min with C. nardus, and 54, 96 and 72 min with S. aromaticum, respectively. Conclusions: The percentage repellency increased when the concentration of essential oils increased. In contrast, biting rates decreased when the concentration of essential oils increased.C. citratus exhibited high efficiency for the protection time and the percentage of biting deterrent against all of 3 mosquito species.

  14. Reduction of malaria transmission to Anopheles mosquitoes with a six-dose regimen of co-artemether.

    Directory of Open Access Journals (Sweden)

    Colin J Sutherland

    2005-04-01

    Full Text Available BACKGROUND: Resistance of malaria parasites to chloroquine (CQ and sulphadoxine-pyrimethamine (SP is increasing in prevalence in Africa. Combination therapy can both improve treatment and provide important public health benefits if it curbs the spread of parasites harbouring resistance genes. Thus, drug combinations must be identified which minimise gametocyte emergence in treated cases, and so prevent selective transmission of parasites resistant to any of the partner drugs. METHODS AND FINDINGS: In a randomised controlled trial, 497 children with uncomplicated falciparum malaria were treated with CQ and SP (three doses and one dose respectively; n = 91, or six doses of artemether in fixed combination with lumefantrine (co-artemether [Coartem, Riamet] (n = 406. Carriage rates of Plasmodium falciparum gametocytes and trophozoites were measured 7, 14, and 28 d after treatment. The infectiousness of venous blood from 29 children carrying P. falciparum gametocytes 7 d after treatment was tested by membrane-feeding of Anopheles mosquitoes. Children treated with co-artemether were significantly less likely to carry gametocytes within the 4 weeks following treatment than those receiving CQ/SP (30 of 378 [7.94%] versus 42 of 86 [48.8%]; p < 0.0001. Carriers in the co-artemether group harboured gametocytes at significantly lower densities, for shorter periods (0.3 d versus 4.2 d; p < 0.0001 and were less infectious to mosquitoes at day 7 (p < 0.001 than carriers who had received CQ/SP. CONCLUSIONS: Co-artemether is highly effective at preventing post-treatment transmission of P. falciparum. Our results suggest that co-artemether has specific activity against immature sequestered gametocytes, and has the capacity to minimise transmission of drug-resistant parasites.

  15. Chemical composition and larvicidal activity of Blumea densiflora essential oils against Anopheles anthropophagus: a malarial vector mosquito.

    Science.gov (United States)

    Zhu, Liang; Tian, Yingjuan

    2011-11-01

    Blumea densiflora, an edible and medicinal plant, is chiefly distributed in Southeast Asia and South Asia. Essential oils extracted by steam distillation from B. densiflora were investigated for their chemical composition and larvicidal activity against Anopheles anthropophagus, the primary vector of malaria in China and other East Asian countries. Totally, 46 compounds were identified by gas chromatography and mass spectroscopy. The major chemical compounds identified were borneol (11.43%), germacrene D (8.66%), β-caryophyllene (6.68%), γ-terpinene (4.35%), sabinene (4.34%), and β-bisabolene (4.24%). A series of concentrations of essential oil (that ranged from 6.25 to 150 ppm) were tested against A. anthropophagus fourth-instar larvae according to WHO recommendation. In general, larval mortality increased as concentration and exposure time increased, indicating a dose-dependent effect, and high insecticidal activity showed that 100% mortality occurred within 6 h at 150 ppm, 10 h at 100 ppm, 30 h at 50 ppm, and 30 h at 25 ppm essential oil concentration. The LC(50) values were 22.32 (after 12 h) and 10.55 ppm (after 24 h), and the LC(90) values were 54.04 (after 12 h) and 33.56 ppm (after 24 h). Pylarvex, the reference standard, had better larvicidal activity, causing 100% mortality within 2 h at 150 ppm and within 6 h at 6.25 ppm. The results clearly reveal that the essential oil of B. densiflora served as a potential, eco-friendly mosquito larvicide against the malarial vector mosquito A. anthropophagus.

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

    Science.gov (United States)

    Gantz, Valentino M; Jasinskiene, Nijole; Tatarenkova, Olga; Fazekas, Aniko; Macias, Vanessa M; Bier, Ethan; James, Anthony A

    2015-12-08

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

  17. Breeding of Anopheles mosquitoes in irrigated areas of South Punjab, Pakistan

    DEFF Research Database (Denmark)

    Herrel, N; Amerasinghe, F P; Ensink, J;

    2001-01-01

    As part of investigations on potential linkages between irrigation and malaria transmission, all surface water bodies in and around three villages along an irrigation distributary in South Punjab, Pakistan, were surveyed for anopheline mosquito larvae (Diptera: Culicidae) from April 1999 to March...

  18. 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.

  19. Mosquito larvicidal properties of Impatiens balsamina (Balsaminaceae) against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera:Culicidae)

    Institute of Scientific and Technical Information of China (English)

    Marimuthu Govindarajan; Mohan Rajeswary

    2014-01-01

    Objective: To assess the larvicidal potential of the crude benzene, chloroform, ethyl acetate and methanol solvent extracts of the medicinal plant Impatiens balsamina against Anopheles stephensi(An. stephensi), Aedes aegypti (Ae. aegypti) and Culex quinquefasciatus (Cx. quinquefasciatus). Methods: Twenty five third instar larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus were exposed to various concentrations and were assayed in the laboratory by using the protocol recommended by WHO. The larval mortality was observed after 24 h of treatment. Results: Among extracts tested, the highest larvicidal activity was observed in leaf methanol extract of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus with the LC50 and LC90 values 98.04, 119.68, 125.06 and 172.93, 210.14, 220.60 mg/L, respectively. Conclusions:From the results it can be concluded that the larvicidal effect of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus make this plant product promising as an alternative to synthetic insecticide in mosquito control programs.

  20. Mosquito larvicidal properties of Impatiens balsamina (Balsaminaceae against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae

    Directory of Open Access Journals (Sweden)

    Marimuthu Govindarajan

    2014-03-01

    Full Text Available Objective: To assess the larvicidal potential of the crude benzene, chloroform, ethyl acetate and methanol solvent extracts of the medicinal plant Impatiens balsamina against Anopheles stephensi (An. stephensi, Aedes aegypti (Ae. aegypti and Culex quinquefasciatus (Cx. quinquefasciatus. Methods: Twenty five third instar larvae of An. stephensi, Ae. aegypti and Cx. quinquefasciatus were exposed to various concentrations and were assayed in the laboratory by using the protocol recommended by WHO. The larval mortality was observed after 24 h of treatment. Results: Among extracts tested, the highest larvicidal activity was observed in leaf methanol extract of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus with the LC50 and LC90 values 98.04, 119.68, 125.06 and 172.93, 210.14, 220.60 mg/L, respectively. Conclusions: From the results it can be concluded that the larvicidal effect of Impatiens balsamina against An. stephensi, Ae. aegypti and Cx. quinquefasciatus make this plant product promising as an alternative to synthetic insecticide in mosquito control programs.

  1. Mecanismos de invasión del esporozoíto de Plasmodium en el mosquito vector Anopheles

    Directory of Open Access Journals (Sweden)

    Lilian M. Spencer

    2016-09-01

    Full Text Available La Malaria o Paludismo es una de las enfermedades tropicales considerada un problema de salud pública a nivel mundial por la OMS. Plasmodium es un protozoario cuyo vector es la hembra del mosquito Anopheles. En este vector se cumplen dos procesos fundamentales en el ciclo de vida del parásito, como son la reproducción sexual, con la formación de un cigoto móvil llamado ooquineto como producto de la fertilización entre los gametos; y la invasión del epitelio del estómago y formación del ooquiste. El estadio producto de esta esporogonia son los esporozoítos (reproducción asexual que se dirigen a las glándulas salivales; y es el infectivo para el mamífero. El esporozoíto es el responsable de establecer la enfermedad en su hospedador vertebrado y por lo tanto los procesos de invasión de este a las glándulas salivales del mosquito es uno de los puntos fundamentales de estudio. Nosotros presentamos una revisión acerca de los mecanismos de invasión del parásito dentro del vector mosquito y las proteínas más importantes que median este proceso. Uno de los aspectos más estudiados en las investigaciones en malaria ha sido determinar la antigenicidad de dichas proteínas en esta parte del ciclo con el fin de ser usadas en el diseño de vacunas. Entre ellas, algunas de las más estudiadas son: P230, P48/45, P28, P25, CTRP, CS, TRAP, WARP y SOAP las cuales han sido consideradas en las estrategias para inhibir el desarrollo del parásito, mejor conocidas como vacunas de bloqueo de trasmisión por el vector. Por lo tanto, presentamos algunas de las estrategias en el diseño de vacunas, basado en las proteínas implicadas en los estadios desarrollados dentro del vector.

  2. Ammonium sulphate fertiliser increases larval populations of Anopheles arabiensis and culicine mosquitoes in rice fields.

    Science.gov (United States)

    Mutero, C M; Ng'ang'a, P N; Wekoyela, P; Githure, J; Konradsen, F

    2004-01-01

    Field experiments were conducted in central Kenya, to study the effect of ammonium sulphate fertiliser ((NH(4))(2)SO(4)) on mosquito larval populations in rice fields. The experiments used a complete randomised block design having four blocks with two experimental ponds per block, and the fertiliser and control treatments allocated randomly among the ponds. Student's two-sample unpaired t-test was used to test for the significance of differences between the relative counts of larvae in fertiliser and control treatments. The results showed a significant overall increase in the larval populations of An. arabiensis (Price fields, thereby making them visually more attractive for egg-laying by An. arabiensis and culicine mosquitoes.

  3. Molecular docking and simulation studies of gustatory receptor of Aedes aegypti: A potent drug target to distract host-seeking behaviour in mosquitoes

    OpenAIRE

    Krishna Kant Gupta; Guneswar Sethi; Manikandan Jayaraman

    2016-01-01

    Background & objectives: It is well reported that exhaled CO 2 and skin odour from human being assist female mosquitoes to locate human host. Basically, the receptors for this activity are expressed in cpA neurons. In both Aedes aegypti and Anopheles gambiae, this CO 2-sensitive olfactory neuron detects myriad number of chemicals present in human skin. Therefore, manipulation of gustatory receptors housing these neurons may serve as important targets for behavioural intervention. The study wa...

  4. Characterization of the doublesex gene within the Culex pipiens complex suggests regulatory plasticity at the base of the mosquito sex determination cascade

    OpenAIRE

    Price, Dana C.; Egizi, Andrea; Dina M Fonseca

    2015-01-01

    Background The doublesex gene controls somatic sexual differentiation of many metazoan species, including the malaria mosquito Anopheles gambiae and the dengue and yellow fever vector Aedes aegypti (Diptera: Culicidae). As in other studied dipteran dsx homologs, the gene maintains functionality via evolutionarily conserved protein domains and sex-specific alternative splicing. The upstream factors that regulate splicing of dsx and the manner in which they do so however remain variable even am...

  5. 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.

  6. Genetic Study of Propoxur Resistance—A Carbamate Insecticide in the Malaria Mosquito, Anopheles stephensi Liston

    Directory of Open Access Journals (Sweden)

    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. Water management for controlling the breeding of Anopheles mosquitoes in rice irrigation schemes in Kenya

    DEFF Research Database (Denmark)

    Mutero, C M; Blank, H; Konradsen, F

    2000-01-01

    between the 4th and 1st instar larvae in the subplots was only 0.08, indicating very low survival rates. In contrast, the 4th/1st instar ratio for subplots with other water management regimes ranged between 0.27 and 0.68, suggesting a correspondingly higher survival than observed with intermittent...... differences among subplots with different water regimes. The average yield per hectare ranged from 4.8-5.3 metric tonnes. The average daily water percolation/seepage rate was 3.6 mm and did not significantly differ among different water management regimes. Further research is necessary to, among other things......An experiment to assess the impact of intermittent irrigation on Anopheles larval populations, rice yields and water use was conducted in the Mwea rice irrigation scheme in Kenya. Four water regimes including intermittent irrigation were tested in a complete randomized block experimental design...

  8. [The distribution of the mosquitoes of the Anopheles maculipennis complex (Diptera, Culicidae, Anophelinae) in Central Asia].

    Science.gov (United States)

    Zvantsov, A B; Gordeev, M I; Goriacheva, I I; Ezhov, M N

    2014-01-01

    The polymerase chain reaction-restriction fragment length polymorphism method, cytogenetic analysis, and investigation of egg exochorion have indicated that three representatives of the Anopheles maculipennis complex (subgenus Anopheles): An artemievi Gordeev et al., An. messeae Falleroni, and An. marinius Shingarev. An. messeae is a European-Siberian species that has extended the southern border of its habitat and has been distributed in the south of Kazakhstan and in the north of Kyrgyzstan. In, Kyrgyzstan, An. messeae inhabiting the plains of Europe and Siberia is encountered rather high up in the mountains: the highest point where this species is found is at 1,879 m above sea level. An. artemievi is present in the highland and piedmont regions of Central Asia (Uzbekistan, Kyrgyzstan, southern Kazakhstan, and northern Tajikistan) and in the intermountain basins (Naryn and Fergana ones). The single finding of this species is in south-eastern Turkmenistan. On the contrary, An. martinius tends to be in the plains and occurs in north-eastern Turkmenistan, Karakalpakstan, and Kazakhstan (Kzyl-Orda). On the other hand, a population of this species is found in proximity to the foothills of the Gissar Range in the east of Uzbekistan. An.maculipennis s.str. is not seen in Central Asia. Early evidence for the presence of both An. maculipennis s.str. and An. martinius in Kopet Dag (Southern Turkmenistan) is rather questionable. It is not improbable that these data are appropriate for either the newly described species An.persiensis or the scientifically new representative of the An. maculipennis complex.

  9. Composition of human skin microbiota affects attractiveness to malaria mosquitoes.

    Directory of Open Access Journals (Sweden)

    Niels O Verhulst

    Full Text Available The African malaria mosquito Anopheles gambiae sensu stricto continues to play an important role in malaria transmission, which is aggravated by its high degree of anthropophily, making it among the foremost vectors of this disease. In the current study we set out to unravel the strong association between this mosquito species and human beings, as it is determined by odorant cues derived from the human skin. Microbial communities on the skin play key roles in the production of human body odour. We demonstrate that the composition of the skin microbiota affects the degree of attractiveness of human beings to this mosquito species. Bacterial plate counts and 16S rRNA sequencing revealed that individuals that are highly attractive to An. gambiae s.s. have a significantly higher abundance, but lower diversity of bacteria on their skin than individuals that are poorly attractive. Bacterial genera that are correlated with the relative degree of attractiveness to mosquitoes were identified. The discovery of the connection between skin microbial populations and attractiveness to mosquitoes may lead to the development of new mosquito attractants and personalized methods for protection against vectors of malaria and other infectious diseases.

  10. 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.

  11. Insecticidal potency of Aspergillus terreus against larvae and pupae of three mosquito species Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti.

    Science.gov (United States)

    Ragavendran, Chinnasamy; Natarajan, Devarajan

    2015-11-01

    Microbial control agents offer alternatives to chemical pest control, as they can be more selective than chemical insecticides. The present study evaluates the mosquito larvicidal and pupicidal potential of fungus mycelia using ethyl acetate and methanol solvent extracts produced by Aspergillus terreus against Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti. The A. terreus mycelia were extracted after 15 days from Sabouraud dextrose broth medium. The ethyl acetate extracts showed lethal concentration that kills 50% of the exposed larvae (LC50) and lethal concentration that kills 90% of the exposed larvae (LC90) values of the first, second, third, and fourth instar larvae of An. stephensi (LC50 = 97.410, 102.551, 29.802, and 8.907; LC90 = 767.957, 552.546, 535.474, and 195.677 μg/ml), Cx. quinquefasciatus (LC50 = 89.584, 74.689, 68.265, and 67.40; LC90 = 449.091, 337.355, 518.793, and 237.347 μg/ml), and Ae. aegypti (LC50 = 83.541, 84.418, 80.407, and 95.926; LC90 = 515.464, 443.167, 387.910, and 473.998 μg/ml). Pupicidal activity of mycelium extracts was tested against An. stephensi (LC50 = 25.228, LC90 = 140.487), Cx. quinquefasciatus (LC50 = 54.525, LC90 = 145.366), and Ae. aegypti (LC50 = 10.536, LC90 = 63.762 μg/ml). At higher concentration (500 μg/ml), mortality starts within the first 6 h of exposure. One hundred percent mortality occurs at 24-h exposure. The overall result observed that effective activity against selected mosquito larvae and pupae after 24 h was a dose and time-dependent activity. These ensure that the resultant mosquito population reduction is substantial even where the larvicidal and pupicidal potential is minimal. The FTIR spectra of ethyl acetate extract reflect prominent peaks (3448.32, 3000.36, 2914.59, 2118.73, 1668.21, 1436.87, 1409.02, 954.33, 901.13, and 704.67 cm(-1)). The spectra showed a sharp absorption band at 1314.66 cm(-1) assigned to wagging vibration of

  12. Ability of TEP1 in intestinal flora to modulate natural resistance of Anopheles dirus.

    Science.gov (United States)

    Wang, Yanyan; Wang, Ying; Zhang, Jingru; Xu, Wenyue; Zhang, Jian; Huang, Fu Sheng

    2013-08-01

    Blocking transmission of malaria is a reliable way to control and eliminate infection. However, in-depth knowledge of the interaction between Plasmodium and mosquito is needed. Studies suggest that innate immunity is the main mechanism inhibiting development of malaria parasites in the mosquito. Recent studies have found that use of antibiotics that inhibit the mosquito gut flora can reduce the immune response of Anopheles gambiae, thereby contributing to the development of malaria parasites. In our study, we used the non susceptible model of Anopheles dirus-Plasmodium yoelii to explore the effect of Anopheles intestinal flora on the natural resistance of A. dirus to P. yoelii. We found that in mosquitoes infected with Plasmodium, the intestinal flora can regulate expression of thioester-containing protein (TEP1) via an RNAi gene-silencing approach. Our results suggest that in the absence of TEP1, the natural microbiota cannot suppress the development of P. yoelii in A. dirus. This suggests that AdTEP1 plays an important role in the resistance of A. dirus to P. yoelii. The intestinal flora may modulate the development of P. yoelii in A. dirus by regulating TEP1 expression.

  13. Evaluating the lethal and pre-lethal effects of a range of fungi against adult Anopheles stephensi mosquitoes

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    Blanford Simon

    2012-11-01

    Full Text Available Abstract Background Insecticide resistance is seriously undermining efforts to eliminate malaria. In response, research on alternatives to the use of chemical insecticides against adult mosquito vectors has been increasing. Fungal entomopathogens formulated as biopesticides have received much attention and have shown considerable potential. This research has necessarily focused on relatively few fungal isolates in order to ‘prove concept’. Further, most attention has been paid to examining fungal virulence (lethality and not the other properties of fungal infection that might also contribute to reducing transmission potential. Here, a range of fungal isolates were screened to examine variation in virulence and how this relates to additional pre-lethal reductions in feeding propensity. Methods The Asian malaria vector, Anopheles stephensi was exposed to 17 different isolates of entomopathogenic fungi belonging to species of Beauveria bassiana, Metarhizium anisopliae, Metarhizium acridum and Isaria farinosus. Each isolate was applied to a test substrate at a standard dose rate of 1×109 spores ml-1 and the mosquitoes exposed for six hours. Subsequently the insects were removed to mesh cages where survival was monitored over the next 14 days. During this incubation period the mosquitoes’ propensity to feed was assayed for each isolate by offering a feeding stimulant at the side of the cage and recording the number probing. Results and conclusions Fungal isolates showed a range of virulence to A. stephensi with some causing >80% mortality within 7 days, while others caused little increase in mortality relative to controls over the study period. Similarly, some isolates had a large impact on feeding propensity, causing >50% pre-lethal reductions in feeding rate, whereas other isolates had very little impact. There was clear correlation between fungal virulence and feeding reduction with virulence explaining nearly 70% of the variation in

  14. Microbial larvicides for malaria control in The Gambia

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    Green Clare

    2007-06-01

    Full Text Available Abstract Background Mosquito larval control may prove to be an effective tool for incorporating into integrated vector management (IVM strategies for reducing malaria transmission. Here the efficacy of microbial larvicides against Anopheles gambiae s.l. was tested in preparation for a large-scale larviciding programme in The Gambia. Methods The impact of water-dispersible (WDG and corn granule (CG formulations of commercial Bacillus sphaericus strain 2362 (Bs; VectoLex® and Bacillus thuringiensis var.israelensis strain AM65-52 (Bti; VectoBac® on larval development were tested under laboratory and field conditions to (1 identify the susceptibility of local vectors, (2 evaluate the residual effect and re-treatment intervals, (3 test the effectiveness of the microbials under operational application conditions and (4 develop a method for large-scale application. Results The major malaria vectors were highly susceptible to both microbials. The lethal concentration (LC to kill 95% of third instar larvae of Anopheles gambiae s.s. after 24 hours was 0.023 mg/l (14.9 BsITU/l for Bs WDG and 0.132 mg/l (396 ITU/l for Bti WDG. In general Bs had little residual effect under field conditions even when the application rate was 200 times greater than the LC95. However, there was a residual effect up to 10 days in standardized field tests implemented during the dry season. Both microbials achieved 100% mortality of larvae 24–48 hours post-application but late instar larvae were detected 4 days after treatment. Pupae development was reduced by 94% (95% Confidence Interval = 90.8–97.5% at weekly re-treatment intervals. Field tests showed that Bs had no residual activity against anopheline larvae. Both microbials provided complete protection when applied weekly. The basic training of personnel in identification of habitats, calibration of application equipment and active larviciding proved to be successful and achieved full coverage and control of mosquito

  15. Evaluation of some aromatic plant extracts for mosquito larvicidal potential against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi.

    Science.gov (United States)

    Jayaraman, M; Senthilkumar, A; Venkatesalu, V

    2015-04-01

    In the present investigation, larvicidal potential of hexane, choloroform, ethyl acetate, acetone, and methanol extracts of seven aromatic plants, viz., Blumea mollis, Chloroxylon swietenia, Clausena anisata, Feronia limnonia, Lantana camera, Plectranthus amboinicus, and Tagetes erecta were screened against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. The larval mortality was observed after 12 and 24 h of exposure period. The results revealed that all the extracts showed varied levels of larvicidal activity against the mosquito species tested. However, the ethyl acetate extract of Chloroxylon swietenia showed the remarkable larvicidal activity against C. quinquefasciatus, Ae. aegypti, and An. stephensi. After 12 h of exposure period, the larvicidal activity was LC50 = 194.22 and LC90 = 458.83 ppm (C. quinquefasciatus), LC50 = 173.04 and LC90 = 442.73 ppm (Ae. aegypti), and LC50 = 167.28 and LC90 = 433.07 ppm (An. stephensi), and the larvicidal activity after 24-h exposure period was LC50 = 94.12 and LC90 = 249.83 ppm (C. quinquefasciatus), LC50 = 80.58 and LC90 = 200.96 ppm (Ae. aegypti), and LC50 = 76.24 and LC90 = 194.51 ppm (An. stephensi). The larvicidal potential of other plant extracts were in order of ethyl acetate extract of Clausena anisata > methanol extract of P. amboinicus > acetone extract of F. limonia > methanol extract of T. erecta > methanol extract of B. mollis > and methanol extract of L. camera. The results of the present study offer a possible way for further investigations to find out the active molecule responsible for the activity.

  16. Suppressor of hairy-wing, modifier of mdg4 and centrosomal protein of 190 gene orthologues of the gypsy insulator complex in the malaria mosquito, Anopheles stephensi.

    Science.gov (United States)

    Carballar-Lejarazú, R; Brennock, P; James, A A

    2016-08-01

    DNA insulators organize independent gene regulatory domains and can regulate interactions amongst promoter and enhancer elements. They have the potential to be important in genome enhancing and editing technologies because they can mitigate chromosomal position effects on transgenes. The orthologous genes of the Anopheles stephensi putative gypsy-like insulator protein complex were identified and expression characteristics studied. These genes encode polypeptides with all the expected protein domains (Cysteine 2 Histidine 2 (C2H2) zinc fingers and/or a bric-a-brac/poxvirus and zinc finger). The mosquito gypsy transcripts are expressed constitutively and are upregulated in ovaries of blood-fed females. We have uncovered significant experimental evidence that the gypsy insulator protein complex is widespread in vector mosquitoes.

  17. 传疟按蚊抗药性研究进展%Research advance on insecticide resistance of malaria vector mosquito Anopheles

    Institute of Scientific and Technical Information of China (English)

    秦茜; 闫桂云; 陈晓光

    2014-01-01

    In May 2010,the Ministry of Health and Chinese Government issued Action Plan of China Malaria Elimination (2010-2020),aiming at complete elimination of malaria in P.R.China by 2020.Vector control is a fundamental element of the existing global strategy to fight malaria.Anopheles is the main malaria vector mosquito.However,rapidly increasing insecticide resistance of mosquitoes threatens current malaria vector control efforts.In order to understand current status of mosquito resistance to insecticide and resistance mechanisms of the malaria vector,the current status of insecticide resistance in malaria vector mosquito Anopheles,the resistance mechanisms and the detection methods on resistance were reviewed in this paper.%我国卫生部于2010年5月印发《中国消除疟疾行动计划(2010-2020年)》,提出在2020年,将全面彻底消除疟疾.控制及消除疟疾的关键在于传播媒介的控制,而按蚊为疟疾主要传播媒介,传疟媒介对杀虫剂的抗药性直接导致了疟疾发病的死灰复燃.为了全面了解疟疾蚊媒的抗药性现状和产生机制,该文对国内外传疟按蚊抗药性现状、产生机制和检测方法进行综述.

  18. Determination of nitric oxide metabolites, nitrate and nitrite, in Anopheles culicifacies mosquito midgut and haemolymph by anion exchange high-performance liquid chromatography: plausible mechanism of refractoriness

    OpenAIRE

    Adak Tridibesh; Raghavendra Kamaraju; Sharma Arun; Dash Aditya P

    2008-01-01

    Abstract Background The diverse physiological and pathological role of nitric oxide in innate immune defenses against many intra and extracellular pathogens, have led to the development of various methods for determining nitric oxide (NO) synthesis. NO metabolites, nitrite (NO2-) and nitrate (NO3-) are produced by the action of an inducible Anopheles culicifacies NO synthase (AcNOS) in mosquito mid-guts and may be central to anti-parasitic arsenal of these mosquitoes. Method While exploring a...

  19. Potential topical natural repellent against Ae. aegypti, Culex sp. and Anopheles sp. mosquitoes

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    Dewi Nur Hodijah

    2014-08-01

    Full Text Available AbstrakLatar belakang:Minyak atsiri daun sirih diketahui mempunyai daya proteksi. Dibuatkan losion berdasarkan pengantar sediaan farmasi yang ditambahkan minyak atsiri daun nilam. Sediaan losion dipilih agar dapat menempel lebih lama di permukaan kulit. Tujuan penelitian ini untuk membandingkan daya proteksi antara losion dengan penambahan minyak nilam dan losion tanpa penambahan minyak nilam dibandingkan daya proteksi dengan DEET. Metode: Penelitian ini merupakan penelitian eksperimental laboratorium. Semua nyamuk uji berasal dari insektarium laboratorium penelitian kesehatan Loka litbang P2B2 Ciamis. Konsentrasi minyak atsiri daun sirih dalam losion adalah 4%; konsentrasi minyak nilam sebagai zat pengikat adalah 0,4%. Formula yang digunakan yaitu formula dasar yang ada pada pengantar sediaan farmasi. Uji repelensi dilakukan dengan menggunakan metoda yang direkomendasikan oleh Komisi pestisida.Hasil: Dihasilkan formulasi losion yang stabil dan masih memenuhi standar formulasi sediaan. Berdasarkan hasil, diperoleh data bahwa DEET dan losion hasil modifikasi memiliki rata-rata daya proteksi di atas 90% selama 6 jam terhadap nyamuk Ae.aegypti dan Culex sp. Kesimpulan: Penambahan minyak nilam pada losion sirih dapat meningkatkan daya proteksi terhadap hinggapan nyamuk Ae. aegypti dan Culex sp. (Health Science Indones 2014;1:44-8Kata kunci:repelen alamiah, minyak atsiri, daun sirih, daun nilam, Ae. aegypti, Culex sp.AbstractBackground: Betel leaf essential oil lotion has been known to have insect repellent properties. A lotion was made based on a pharmaceutical formula from a monograph where patchouli leaf essential oil was added. A lotion preparation was intended to enhance adherence of the formula on the surface of the skin. The purpose of this study was to compare protection percentage of lotion with patchouli oil and without patchouli oil lotion compared to DEET.Methods: This study is an experimental laboratory-based research. All mosquitoes

  20. 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.

  1. Deep sequencing revealed molecular signature of horizontal gene transfer of plant like transcripts in the mosquito Anopheles culicifacies: an evolutionary puzzle [version 1; referees: 2 approved

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    Punita Sharma

    2015-12-01

    Full Text Available In prokaryotes, horizontal gene transfer (HGT has been regarded as an important evolutionary drive to acquire and retain beneficial genes for their survival in diverse ecologies. However, in eukaryotes, the functional role of HGTs remains questionable, although current genomic tools are providing increased evidence of acquisition of novel traits within non-mating metazoan species. Here, we provide another transcriptomic evidence for the acquisition of massive plant genes in the mosquito, Anopheles culicifacies. Our multiple experimental validations including genomic PCR, RT-PCR, real-time PCR, immuno-blotting and immuno-florescence microscopy, confirmed that plant like transcripts (PLTs are of mosquito origin and may encode functional proteins. A comprehensive molecular analysis of the PLTs and ongoing metagenomic analysis of salivary microbiome provide initial clues that mosquitoes may have survival benefits through the acquisition of nuclear as well as chloroplast encoded plant genes. Our findings of PLTs further support the similar questionable observation of HGTs in other higher organisms, which is still a controversial and debatable issue in the community of evolutionists. We believe future understanding of the underlying mechanism of the feeding associated molecular responses may shed new insights in the functional role of PLTs in the mosquito.

  2. Cultured skin microbiota attracts malaria mosquitoes

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    Takken Willem

    2009-12-01

    Full Text Available Abstract Background Host-seeking of the African malaria mosquito, Anopheles gambiae sensu stricto, is guided by human odours. The precise nature of the odours, and the composition of attractive blends of volatiles, remains largely unknown. Skin microbiota plays an important role in the production of human body odours. It is hypothesized that host attractiveness and selection of An. gambiae is affected by the species composition, density, and metabolic activity of the skin microbiota. A study is presented in which the production and constituency of volatile organic compounds (VOCs by human skin microbiota is examined and the behavioural responses of An. gambiae to VOCs from skin microbiota are investigated. Methods Blood agar plates incubated with skin microbiota from human feet or with a reference strain of Staphylococcus epidermidis were tested for their attractiveness to An. gambiae in olfactometer bioassays and indoor trapping experiments. Entrained air collected from blood agar plates incubated with natural skin microbiota or with S. epidermidis were analysed using GC-MS. A synthetic blend of the compounds identified was tested for its attractiveness to An. gambiae. Behavioural data were analysed by a χ2-test and GLM. GC-MS results were analysed by fitting an exponential regression line to test the effect of the concentration of bacteria. Results More An. gambiae were caught with blood agar plates incubated with skin bacteria than with sterile blood agar plates, with a significant effect of incubation time and dilution of the skin microbiota. When bacteria from the feet of four other volunteers were tested, similar effects were found. Fourteen putative attractants were found in the headspace of the skin bacteria. A synthetic blend of 10 of these was attractive to An. gambiae. Conclusions The discovery that volatiles produced by human skin microorganisms in vitro mediate An. gambiae host-seeking behaviour creates new opportunities for the

  3. The distribution and bionomics of anopheles malaria vector mosquitoes in Indonesia.

    Science.gov (United States)

    Elyazar, Iqbal R F; Sinka, Marianne E; Gething, Peter W; Tarmidzi, Siti N; Surya, Asik; Kusriastuti, Rita; Winarno; Baird, J Kevin; Hay, Simon I; Bangs, Michael J

    2013-01-01

    Malaria remains one of the greatest human health burdens in Indonesia. Although Indonesia has a long and renowned history in the early research and discoveries of malaria and subsequently in the successful use of environmental control methods to combat the vector, much remains unknown about many of these mosquito species. There are also significant gaps in the existing knowledge on the transmission epidemiology of malaria, most notably in the highly malarious eastern half of the archipelago. These compound the difficulty of developing targeted and effective control measures. The sheer complexity and number of malaria vectors in the country are daunting. The difficult task of summarizing the available information for each species and/or species complex is compounded by the patchiness of the data: while relatively plentiful in one area or region, it can also be completely lacking in others. Compared to many other countries in the Oriental and Australasian biogeographical regions, only scant information on vector bionomics and response to chemical measures is available in Indonesia. That information is often either decades old, geographically patchy or completely lacking. Additionally, a large number of information sources are published in Dutch or Indonesian language and therefore less accessible. This review aims to present an updated overview of the known distribution and bionomics of the 20 confirmed malaria vector species or species complexes regarded as either primary or secondary (incidental) malaria vectors within Indonesia. This chapter is not an exhaustive review of each of these species. No attempt is made to specifically discuss or resolve the taxonomic record of listed species in this document, while recognizing the ever evolving revisions in the systematics of species groups and complexes. A review of past and current status of insecticide susceptibility of eight vector species of malaria is also provided.

  4. Mosquito larvicidal properties of silver nanoparticles synthesized using Heliotropium indicum (Boraginaceae) against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus (Diptera: Culicidae).

    Science.gov (United States)

    Veerakumar, Kaliyan; Govindarajan, Marimuthu; Rajeswary, Mohan; Muthukumaran, Udaiyan

    2014-06-01

    Mosquitoes transmit dreadful diseases to human beings wherein biological control of these vectors using plant-derived molecules would be an alternative to reduce mosquito population. In the present study activity of aqueous leaf extract and silver nanoparticles (AgNPs) synthesized using Helitropium indicum plant leaves against late third instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. The range of varying concentrations of synthesized AgNPs (8, 16, 24, 32, and 40 μg/mL) and aqueous leaf extract (30, 60, 90, 120, and 150 μg/mL) were tested against the larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The synthesized AgNPs from H. indicum 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, energy-dispersive X-ray spectroscopy analysis, transmission electron microscopy, and histogram. The synthesized AgNPs showed larvicidal effects after 24 h of exposure. Considerable mortality was evident after the treatment of H. indicum for all three important vector mosquitoes. The LC50 and LC90 values of H. indicum aqueous leaf extract appeared to be effective against A. stephensi (LC50, 68.73 μg/mL; LC90, 121.07 μg/mL) followed by A. aegypti (LC50, 72.72 μg/mL; LC90, 126.86 μg/mL) and C. quinquefasciatus (LC50, 78.74 μg/mL; LC90, 134.39 μg/mL). Synthesized AgNPs against the vector mosquitoes of A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC50 and LC90 values: A. stephensi had LC50 and LC90 values of 18.40 and 32.45 μg/mL, A. aegypti had LC50 and LC90 values of 20.10 and 35.97 μg/mL, and C. quinquefasciatus had LC50 and LC90 values of 21.84 and 38.10 μg/mL. No mortality was observed in the control. These results suggest that the leaf aqueous extracts of H. indicum and green synthesis of silver nanoparticles have the

  5. Efficacy of AquatainTM, a monomolecular surface film, against the malaria vectors Anopheles stephensi and An. gambiae s.s. in the laboratory

    NARCIS (Netherlands)

    Bukhari, S.T.; Knols, B.G.J.

    2009-01-01

    Monomolecular films are used for mosquito control because of their asphyxiating effect on larvae and pupae. Compared with other films, Aquatain mosquito formulation (AMFTM) has an improved spreading ability and flexibility on a water surface. In the laboratory, AMFTM showed larvicidal, pupicidal, an

  6. Mosquito cell line glycoproteins: an unsuitable model system for the Plasmodium ookinete-mosquito midgut interaction?

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    Wilkins Simon

    2010-03-01

    Full Text Available Abstract Background Mosquito midgut glycoproteins may act as key recognition sites for the invading malarial ookinete. Effective transmission blocking strategies require the identification of novel target molecules. We have partially characterised the surface glycoproteins of two cell lines from two mosquito species; Anopheles stephensi and Anopheles gambiae, and investigated the binding of Plasmodium berghei ookinetes to carbohydrate ligands on the cells. Cell line extracts were run on SDS-PAGE gels and carbohydrate moieties determined by blotting against a range of biotinylated lectins. In addition, specific glycosidases were used to cleave the oligosaccharides. Results An. stephensi 43 and An. gambiae 55 cell line glycoproteins expressed oligosaccharides containing oligomannose and hybrid oligosaccharides, with and without α1-6 core fucosylation; N-linked oligosaccharides with terminal Galβ1-3GalNAc or GalNAcβ1-3Gal; O-linked α/βGalNAc. An. stephensi 43 cell line glycoproteins also expressed N-linked Galβ1-4R and O-linked Galβ1-3GalNAc. Although P. berghei ookinetes bound to both mosquito cell lines, binding could not be inhibited by GlcNAc, GalNAc or Galactose. Conclusions Anopheline cell lines displayed a limited range of oligosaccharides. Differences between the glycosylation patterns of the cell lines and mosquito midgut epithelial cells could be a factor why ookinetes did not bind in a carbohydrate inhibitable manner. Anopheline cell lines are not suitable as a potential model system for carbohydrate-mediated adhesion of Plasmodium ookinetes.

  7. Spectral and HRTEM analyses of Annona muricata leaf extract mediated silver nanoparticles and its Larvicidal efficacy against three mosquito vectors Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti.

    Science.gov (United States)

    Santhosh, Shanthi Bhupathi; Ragavendran, Chinnasamy; Natarajan, Devarajan

    2015-12-01

    Mosquitoes transmit various diseases which mainly affect the human beings and every year cause millions of deaths globally. Currently available chemical and synthetic mosquitocidal agents pose severe side effects, pollute the environment vigorously, and become resistance. There is an urgent need to identify and develop the cost effective, compatible and eco-friendly product for mosquito control. The present study was aimed to find out the larvicidal potential of aqueous crude extract and green synthesized silver nanoparticles (AgNPs) from Annona muricata leaves were tested against fourth instar larvae of three important mosquitoes i.e. Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti using different concentrations of AgNPs (10, 20, 30, 40 and 50 ppm) and the aqueous leaf extract (100, 200, 300, 400, and 500 ppm) for 24 and 48 h. The maximum mortality was noticed in AgNPs than aqueous leaf extract of A. muricata against tested mosquitoes with least LC50 values of 37.70, 31.29, and 20.65 ppm (24h) and 546.7, 516.2, and 618.4 ppm (48 h), respectively. All tested concentrations of AgNps exhibited 100% mortality in A. aegypti larvae at 48 hour observation. In addition, the plant mediated AgNPs were characterized by UV-vis spectrum, Fourier transform infrared spectroscopy, particle size analyser, X-ray diffraction, high resonance transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis for confirmation of nanoparticle synthesis. Based on the findings of the study suggests that the use of A. muricata plant mediated AgNPs can act as an alternate insecticidal agents for controlling target mosquitoes.

  8. Chemical composition and larvicidal activity of the essential oil of Plectranthus amboinicus (Lour.) Spreng against Anopheles stephensi: a malarial vector mosquito.

    Science.gov (United States)

    Senthilkumar, Annadurai; Venkatesalu, Venugopalan

    2010-10-01

    Essential oil of Plectranthus amboinicus was studied for its chemical composition and larvicidal potential against the malarial vector mosquito Anopheles stephensi. Totally 26 compounds were identified by GC and GC-MS. The major chemical compounds were carvacrol (28.65%) followed by thymol (21.66%), α-humulene (9.67%), undecanal (8.29%), γ-terpinene (7.76%), ρ-cymene (6.46%), caryophyllene oxide (5.85%), α-terpineol (3.28%) and β-selinene (2.01%). The larvicidal assay was conducted to record the LC(50) and LC(90) values and the larval mortality was observed after 12 and 24 h of exposure period. The LC(50) values of the oil were 33.54 (after 12 h) and 28.37 ppm (after 24 h). The LC(90) values of the oil were 70.27 (after 12 h) and 59.38 ppm (after 24 h). The results of the present study showed that the essential oil of P. amboinicus is one of the inexpensive and eco-friendly sources of natural mosquito larvicidal agent to control/reduce the population of malarial vector mosquito.

  9. Trapping of the malaria vector Anopheles gambiae with odour-baited MM-X traps in semi-field conditions in western Kenya

    NARCIS (Netherlands)

    Njiru, B.N.; Mukabana, W.R.; Takken, W.; Knols, B.G.J.

    2006-01-01

    Background - The successful development of odour-baited trapping systems for mosquitoes depends on the identification of behaviourally active semiochemicals, besides the design and operating principles of such devices. A large variety of 'attractants' has been identified in laboratory investigations

  10. Gravid Anopheles gambiae sensu stricto avoid ovipositing in Bermuda grass hay infusion and it’s volatiles in two choice egg-count bioassays

    OpenAIRE

    Eneh, LK; Okal, MN; Borg-Karlson, AK; Fillinger, U; Lindh, JM

    2016-01-01

    Background A number of mosquito species in the Culex and Aedes genera prefer to lay eggs in Bermuda grass (Cynodon dactylon) hay infusions compared to water alone. These mosquitoes are attracted to volatile compounds from the hay infusions making the infusions effective baits in gravid traps used for monitoring vectors of arboviral and filarial pathogens. Since Bermuda grass is abundant and widespread, it is plausible to explore infusions made from it as a potential low cost bait for outdoor ...

  11. Risk factors for house-entry by culicine mosquitoes in a rural town and satellite villages in The Gambia

    Directory of Open Access Journals (Sweden)

    Jasseh Momodou

    2008-10-01

    Full Text Available Abstract Background Screening doors, windows and eaves of houses should reduce house entry by eusynanthropic insects, including the common African house mosquito Culex pipiens quinquefasciatus and other culicines. In the pre-intervention year of a randomized controlled trial investigating the protective effects of house screening against mosquito house entry, a multi-factorial risk factor analysis study was used to identify factors influencing house entry by culicines of nuisance biting and medical importance. These factors were house location, architecture, human occupancy and their mosquito control activities, and the number and type of domestic animals within the compound. Results 40,407 culicines were caught; the dominant species were Culex thalassius, Cx. pipiens s.l., Mansonia africanus, M. uniformis and Aedes aegypti. There were four times more Cx. pipiens s.l. in Farafenni town (geometric mean/trap/night = 8.1, 95% confidence intervals, CIs = 7.2–9.1 than in surrounding villages (2.1, 1.9–2.3, but over five times more other culicines in the villages (25.1, 22.1–28.7 than in town (4.6, 4.2–5.2. The presence of Cx. pipiens s.l. was reduced in both settings if the house had closed eaves (odds ratios, OR town = 0.62, 95% CIs = 0.49–0.77; OR village = 0.49, 0.33–0.73, but increased per additional person in the trapping room (OR town = 1.16, 1.09–1.24; OR village = 1.10, 1.02–1.18. In the town only, Cx. pipiens s.l. numbers were reduced if houses had a thatched roof (OR = 0.70, 0.51–0.96, for each additional cow tethered near the house (OR = 0.73, 0.65–0.82 and with increasing distance from a pit latrine (OR = 0.97, 0.95–0.99. In the villages a reduction in Cx. pipiens s.l. numbers correlated with increased horses in the compound (OR = 0.90, 0.82–0.99. The presence of all other culicines was reduced in houses with closed eaves (both locations, with horses tethered outside (village only and with increasing room height

  12. Screening mosquito house entry points as a potential method for integrated control of endophagic filariasis, arbovirus and malaria vectors.

    Directory of Open Access Journals (Sweden)

    Sheila B Ogoma

    Full Text Available BACKGROUND: Partial mosquito-proofing of houses with screens and ceilings has the potential to reduce indoor densities of malaria mosquitoes. We wish to measure whether it will also reduce indoor densities of vectors of neglected tropical diseases. METHODOLOGY: The main house entry points preferred by anopheline and culicine vectors were determined through controlled experiments using specially designed experimental huts and village houses in Lupiro village, southern Tanzania. The benefit of screening different entry points (eaves, windows and doors using PVC-coated fibre glass netting material in terms of reduced indoor densities of mosquitoes was evaluated compared to the control. FINDINGS: 23,027 mosquitoes were caught with CDC light traps; 77.9% (17,929 were Anopheles gambiae sensu lato, of which 66.2% were An. arabiensis and 33.8% An. gambiae sensu stricto. The remainder comprised 0.2% (50 An. funestus, 10.2% (2359 Culex spp. and 11.6% (2664 Mansonia spp. Screening eaves reduced densities of Anopheles gambiae s. l. (Relative ratio (RR = 0.91; 95% CI = 0.84, 0.98; P = 0.01; Mansonia africana (RR = 0.43; 95% CI = 0.26, 0.76; P<0.001 and Mansonia uniformis (RR = 0.37; 95% CI = 0.25, 0.56; P<0.001 but not Culex quinquefasciatus, Cx. univittatus or Cx. theileri. Numbers of these species were reduced by screening windows and doors but this was not significant. SIGNIFICANCE: This study confirms that across Africa, screening eaves protects households against important mosquito vectors of filariasis, Rift Valley Fever and O'Nyong nyong as well as malaria. While full house screening is required to exclude Culex species mosquitoes, screening of eaves alone or fitting ceilings has considerable potential for integrated control of other vectors of filariasis, arbovirus and malaria.

  13. To assess whether indoor residual spraying can provide additional protection against clinical malaria over current best practice of long-lasting insecticidal mosquito nets in The Gambia: study protocol for a two-armed cluster-randomised trial

    Directory of Open Access Journals (Sweden)

    Parker David

    2011-06-01

    Full Text Available Abstract Background Recently, there has been mounting interest in scaling-up vector control against malaria in Africa. It needs to be determined if indoor residual spraying (IRS with DDT will provide significant marginal protection against malaria over current best practice of long-lasting insecticidal nets (LLINs and prompt treatment in a controlled trial, given that DDT is currently the most persistent insecticide for IRS. Methods A 2 armed cluster-randomised controlled trial will be conducted to assess whether DDT IRS and LLINs combined provide better protection against clinical malaria in children than LLINs alone in rural Gambia. Each cluster will be a village, or a group of small adjacent villages; all clusters will receive LLINs and half will receive IRS in addition. Study children, aged 6 months to 13 years, will be enrolled from all clusters and followed for clinical malaria using passive case detection to estimate malaria incidence for 2 malaria transmission seasons in 2010 and 2011. This will be the primary endpoint. Exposure to malaria parasites will be assessed using light and exit traps followed by detection of Anopheles gambiae species and sporozoite infection. Study children will be surveyed at the end of each transmission season to estimate the prevalence of Plasmodium falciparum infection and the prevalence of anaemia. Discussion Practical issues concerning intervention implementation, as well as the potential benefits and risks of the study, are discussed. Trial Registration ISRCTN01738840 - Spraying And Nets Towards malaria Elimination (SANTE

  14. Impact of mating behaviour on the success of malaria control through a single inundative release of transgenic mosquitoes.

    Science.gov (United States)

    Boëte, C; Agusto, F B; Reeves, R G

    2014-04-21

    Transgenic mosquitoes are a potential tool for the control or eradication of insect-vectored diseases. For malaria, one possible strategy relies on the introduction of malaria-refractory transgenes into wild Anopheles mosquito populations that would limit their capacity to transmit the disease. The success of such an approach obviously depends on a variety of factors. By developing a model that integrates both population genetics and epidemiology, we explore how mosquito mating preferences and the cost and efficacy of refractoriness affects the long-term prevalence of malaria in humans subsequent to a single generation inundative release of male transgenic mosquitoes. As may be intuitively expected, mating discrimination by wild-type individuals against transgenic ones generally reduces the probability that transgenes become stably established at a high frequency in mosquito populations. We also show that in circumstances where transgenic individuals exhibit some degree of discrimination against wild-type individuals, this can favour the spread of refractory alleles and lead to a significant reduction in malaria prevalence in the human population (if the efficacy of a dominant refractory mechanism exceeds at least 75%). The existence of such a non-intuitive outcome highlights the practical value of increasing the understanding of Anopheles mating preferences in the wild as a means to harness them in the implementation of population replacement approaches. Potential strategies by which previously described mating preferences of Anopheles gambiae populations could be exploited to manipulate the mate choice of transgenic release stocks are discussed.