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Sample records for aegypti anopheles gambiae

  1. oskar gene expression in the vector mosquitoes, Anopheles gambiae and Aedes aegypti.

    PubMed

    Juhn, J; James, A A

    2006-06-01

    A disease control strategy based on the introduction into mosquito populations of a gene conferring a pathogen-refractory phenotype is currently under investigation. This population replacement approach requires a drive system that will quickly spread and fix antipathogen effector genes in target populations. Modified transposable elements containing the control sequences of developmentally regulated genes may provide the basis for a gene drive system that regulates gene mobilization in a sex- and stage-restrictive manner. Screening of a Drosophila melanogaster database for genes whose products localize exclusively in the future germ cells during early embryonic development resulted in the identification of several candidate genes. The regulatory sequences of these genes could be used to drive transposition. Mosquito orthologous genes of oskar were identified based on sequence homology and characterized further. The tissue- and sex-specific expression profiles and hybridizations in situ show that oskar orthologous transcripts in Anopheles gambiae and Aedes aegypti accumulate in developing oocytes of adult females and localize to the posterior poles of early embryos. These characteristics potentiate the use of the regulatory sequences of mosquito oskar genes for the control of modified transposable elements.

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

    PubMed

    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.

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

    PubMed

    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. PMID:26336230

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

    PubMed Central

    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. PMID:26447952

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

    PubMed

    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. PMID:26447952

  6. Genome-Based Microsatellite Development in the Culex pipiens Complex and Comparative Microsatellite Frequency with Aedes aegypti and Anopheles gambiae

    PubMed Central

    Hickner, Paul V.; deBruyn, Becky; Lovin, Diane D.; Mori, Akio; Behura, Susanta K.; Pinger, Robert; Severson, David W.

    2010-01-01

    Background Mosquitoes in the Culex pipiens complex are among the most medically important vectors for human disease worldwide and include major vectors for lymphatic filariasis and West Nile virus transmission. However, detailed genetic studies in the complex are limited by the number of genetic markers available. Here, we describe methods for the rapid and efficient identification and development of single locus, highly polymorphic microsatellite markers for Cx. pipiens complex mosquitoes via in silico screening of the Cx. quinquefasciatus genome sequence. Methodology/Principal Findings Six lab colonies representing four Cx. pipiens and two Cx. quinquefasciatus populations were utilized for preliminary assessment of 38 putative loci identified within 16 Cx. quinquefasciatus supercontig assemblies (CpipJ1) containing previously mapped genetic marker sequences. We identified and validated 12 new microsatellite markers distributed across all three linkage groups that amplify consistently among strains representing the complex. We also developed four groups of 3–5 microsatellite loci each for multiplex-ready PCR. Field collections from three cities in Indiana were used to assess the multiplex groups for their application to natural populations. All were highly polymorphic (Mean  = 13.0 alleles) per locus and reflected high polymorphism information content (PIC) (Mean  = 0.701). Pairwise FST indicated population structuring between Terre Haute and Fort Wayne and between Terre Haute and Indianapolis, but not between Fort Wayne and Indianapolis. In addition, we performed whole genome comparisons of microsatellite motifs and abundance between Cx. quinquefasciatus and the primary vectors for dengue virus and malaria parasites, Aedes aegypti and Anopheles gambiae, respectively. Conclusions/Significance We demonstrate a systematic approach for isolation and validation of microsatellites for the Cx. pipiens complex by direct screen of the Cx. quinquefasciatus genome

  7. Brain Proteomics of Anopheles gambiae

    PubMed Central

    Dwivedi, Sutopa B.; Muthusamy, Babylakshmi; Kumar, Praveen; Kim, Min-Sik; Nirujogi, Raja Sekhar; Getnet, Derese; Ahiakonu, Priscilla; De, Gourav; Nair, Bipin; Gowda, Harsha; Prasad, T.S. Keshava; Kumar, Nirbhay

    2014-01-01

    Abstract Anopheles gambiae has a well-adapted system for host localization, feeding, and mating behavior, which are all governed by neuronal processes in the brain. However, there are no published reports characterizing the brain proteome to elucidate neuronal signaling mechanisms in the vector. To this end, a large-scale mapping of the brain proteome of An. gambiae was carried out using high resolution tandem mass spectrometry, revealing a repertoire of >1800 proteins, of which 15% could not be assigned any function. A large proportion of the identified proteins were predicted to be involved in diverse biological processes including metabolism, transport, protein synthesis, and olfaction. This study also led to the identification of 10 GPCR classes of proteins, which could govern sensory pathways in mosquitoes. Proteins involved in metabolic and neural processes, chromatin modeling, and synaptic vesicle transport associated with neuronal transmission were predominantly expressed in the brain. Proteogenomic analysis expanded our findings with the identification of 15 novel genes and 71 cases of gene refinements, a subset of which were validated by RT-PCR and sequencing. Overall, our study offers valuable insights into the brain physiology of the vector that could possibly open avenues for intervention strategies for malaria in the future. PMID:24937107

  8. Tools for Anopheles gambiae Transgenesis.

    PubMed

    Volohonsky, Gloria; Terenzi, Olivier; Soichot, Julien; Naujoks, Daniel A; Nolan, Tony; Windbichler, Nikolai; Kapps, Delphine; Smidler, Andrea L; Vittu, Anaïs; Costa, Giulia; Steinert, Stefanie; Levashina, Elena A; Blandin, Stéphanie A; Marois, Eric

    2015-06-01

    Transgenesis is an essential tool to investigate gene function and to introduce desired characters in laboratory organisms. Setting-up transgenesis in non-model organisms is challenging due to the diversity of biological life traits and due to knowledge gaps in genomic information. Some procedures will be broadly applicable to many organisms, and others have to be specifically developed for the target species. Transgenesis in disease vector mosquitoes has existed since the 2000s but has remained limited by the delicate biology of these insects. Here, we report a compilation of the transgenesis tools that we have designed for the malaria vector Anopheles gambiae, including new docking strains, convenient transgenesis plasmids, a puromycin resistance selection marker, mosquitoes expressing cre recombinase, and various reporter lines defining the activity of cloned promoters. This toolbox contributed to rendering transgenesis routine in this species and is now enabling the development of increasingly refined genetic manipulations such as targeted mutagenesis. Some of the reagents and procedures reported here are easily transferable to other nonmodel species, including other disease vector or agricultural pest insects.

  9. Neuropeptides and Peptide Hormones in Anopheles gambiae

    NASA Astrophysics Data System (ADS)

    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.

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

    PubMed Central

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

    2015-01-01

    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. PMID:26220140

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  12. Salivary vasodilators of Aedes triseriatus and Anopheles gambiae (Diptera: Culicidae).

    PubMed

    Ribeiro, J M; Nussenzveig, R H; Tortorella, G

    1994-09-01

    Salivary vasodilators of Aedes aegypti (L.) and Anopheles albimanus (Wiedemann) were characterized previously as a tachykinin peptide and a catechol oxidase/peroxidase activity, respectively. To verify whether these two different vasodilators also were found in other distantly related members of each mosquito genus, we characterized the vasodilators from A. triseriatus and A. gambiae. A. triseriatus salivary gland homogenates produced a reversible, endothelium dependent vasorelaxation of rabbit aortic rings constricted with norepinephrine, and contracted an isolated guinea pig ileum preparation. Additionally, the homogenate had no activity on both smooth muscle preparations when both tissues were desensitized previously by a large dose of substance P. Taken together, these assays suggest the presence of a salivary tachykinin in A. triseriatus. A. gambiae salivary gland homogenates induced a slow vasodilation on both endothelium intact and endotheliumless preparations of aortic rings. A. gambiae homogenates also displayed catechol oxidase and peroxidase activity against o-dianisidine but not against serotonin, indicating the presence of an enzyme system slightly different from A. albimanus. We conclude that the presence of salivary tachykinins or catechol/oxidase is not unique to A. aegypti or A. albimanus. PMID:7966179

  13. Biological variations in Anopheles darlingi and Anopheles gambiae

    PubMed Central

    Giglioli, G.

    1956-01-01

    A remarkable range of variation has been demonstrated in the biting habits of A. darlingi in South America. At the centre of its area of distribution, the species comprises both anthropo-endophilic and zoo-exophilic individuals, but the anthropophilic tendencies increase progressively towards the periphery of its distributional area. Conditions of travel and migration, imposed by the South American rain forest, have probably favoured the development of a specialized anthropophilic strain on a geographical pattern. There is ample evidence of a similar strain evolution in A. gambiae, but on a more confused pattern. Aëdes aegypti also presents an outstanding example of variation in biting habits. A proper understanding of the extent and significance on anthropophilic and zoophilic tendencies is essential for the proper conduct of malaria control campaigns; for the formulation of long-term policies; and for the proper evaluation of the probabilities of resistance to insecticides. PMID:13404433

  14. RNAi Trigger Delivery into Anopheles gambiae Pupae

    PubMed Central

    Regna, Kimberly; Harrison, Rachel M.; Heyse, Shannon A.; Chiles, Thomas C.; Michel, Kristin; Muskavitch, Marc A. T.

    2016-01-01

    RNA interference (RNAi), a naturally occurring phenomenon in eukaryotic organisms, is an extremely valuable tool that can be utilized in the laboratory for functional genomic studies. The ability to knockdown individual genes selectively via this reverse genetic technique has allowed many researchers to rapidly uncover the biological roles of numerous genes within many organisms, by evaluation of loss-of-function phenotypes. In the major human malaria vector Anopheles gambiae, the predominant method used to reduce the function of targeted genes involves injection of double-stranded (dsRNA) into the hemocoel of the adult mosquito. While this method has been successful, gene knockdown in adults excludes the functional assessment of genes that are expressed and potentially play roles during pre-adult stages, as well as genes that are expressed in limited numbers of cells in adult mosquitoes. We describe a method for the injection of Serine Protease Inhibitor 2 (SRPN2) dsRNA during the early pupal stage and validate SRPN2 protein knockdown by observing decreased target protein levels and the formation of melanotic pseudo-tumors in SRPN2 knockdown adult mosquitoes. This evident phenotype has been described previously for adult stage knockdown of SRPN2 function, and we have recapitulated this adult phenotype by SRPN2 knockdown initiated during pupal development. When used in conjunction with a dye-labeled dsRNA solution, this technique enables easy visualization by simple light microscopy of injection quality and distribution of dsRNA in the hemocoel. PMID:27023367

  15. Anopheles gambiae complex along The Gambia river, with particular reference to the molecular forms of An. gambiae s.s

    PubMed Central

    Caputo, Beniamino; Nwakanma, Davis; Jawara, Musa; Adiamoh, Majidah; Dia, Ibrahima; Konate, Lassana; Petrarca, Vincenzo; Conway, David J; della Torre, Alessandra

    2008-01-01

    Background The geographic and temporal distribution of M and S molecular forms of the major Afrotropical malaria vector species Anopheles gambiae s.s. at the western extreme of their range of distribution has never been investigated in detail. Materials and methods Collections of indoor-resting An. gambiae s.l. females were carried out along a ca. 400 km west to east transect following the River Gambia from the western coastal region of The Gambia to south-eastern Senegal during 2005 end of rainy season/early dry season and the 2006 rainy season. Specimens were identified to species and molecular forms by PCR-RFLP and the origin of blood-meal of fed females was determined by ELISA test. Results Over 4,000 An. gambiae s.l. adult females were collected and identified, 1,041 and 3,038 in 2005 and 2006, respectively. M-form was mainly found in sympatry with Anopheles melas and S-form in the western part of the transect, and with Anopheles arabiensis in the central part. S-form was found to prevail in rural Sudan-Guinean savannah areas of Eastern Senegal, in sympatry with An. arabiensis. Anopheles melas and An. arabiensis relative frequencies were generally lower in the rainy season samples, when An. gambiae s.s. was prevailing. No large seasonal fluctuations were observed for M and S-forms. In areas where both M and S were recorded, the frequency of hybrids between them ranged from to 0.6% to 7%. Discussion The observed pattern of taxa distribution supports the hypothesis of a better adaptation of M-form to areas characterized by water-retaining alluvial deposits along the Gambia River, characterized by marshy vegetation, mangrove woods and rice cultivations. In contrast, the S-form seems to be better adapted to free-draining soil, covered with open woodland savannah or farmland, rich in temporary larval breeding sites characterizing mainly the eastern part of the transect, where the environmental impact of the Gambia River is much less profound and agricultural

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

    PubMed

    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. PMID:25417803

  17. Two genes encoding midgut-specific maltase-like polypeptides from Anopheles gambiae.

    PubMed

    Zheng, L; Whang, L H; Kumar, V; Kafatos, F C

    1995-11-01

    Full-length cDNA clones of two genes have been isolated from the African malaria vector mosquito, Anopheles gambiae. These genes, designated Agm1 and Agm2, encode maltase-like polypeptides of 498 and 599 residues, respectively. Deduced amino acid sequences contain a putative signal peptide sequence and four potential glycosylation sites. Agm1 and Agm2 show highest similarities to the Mal1 gene from Aedes aegypti and three clustered maltase genes from Drosophila melanogaster. Both genes are located at position 46D, in the terminal division of the left arm of the third chromosome. Agm2 has very strict tissue and temporal specificity, being expressed exclusively in the adult midgut. The specificity of Agm1 is similar but appears slightly broader; transcripts of this gene are detected at a low level in the pupae, and occasionally in the adult carcass after removal of the midgut.

  18. The problem of exophily in Anopheles gambiae

    PubMed Central

    Gillies, M. T.

    1956-01-01

    Studies of the exophily exhibited by anophelines in a humid coastal area and in an arid inland region of Tanganyika gave very different results. In the former area, catches of A. gambiae were scanty and largely composed of unfed and gravid females, while in the latter, large numbers of mosquitos were caught outside and a great many of them were recently fed females. The differences are attributed primarily to the presence outside of large herds of cattle in the inland region and to their absence near the coast. The reports of exophily from other parts of Africa are also analysed and show that there is much variation in the behaviour of A. gambiae in different regions. Some of the variation can be explained in terms of environmental differences, particularly in the availability at night of different hosts. But some of it may be genetically determined. It is suggested that the main task in this field is twofold: firstly, to establish the existence and nature of the behaviour differences described; secondly, to study in detail the mosquitos that survive as exophilic populations in areas where systematic house-spraying is in operation. PMID:13404431

  19. Landscape Movements of Anopheles gambiae Malaria Vector Mosquitoes in Rural Gambia

    PubMed Central

    Thomas, Christopher J.; Cross, Dónall E.; Bøgh, Claus

    2013-01-01

    Background For malaria control in Africa it is crucial to characterise the dispersal of its most efficient vector, Anopheles gambiae, in order to target interventions and assess their impact spatially. Our study is, we believe, the first to present a statistical model of dispersal probability against distance from breeding habitat to human settlements for this important disease vector. Methods/Principal Findings We undertook post-hoc analyses of mosquito catches made in The Gambia to derive statistical dispersal functions for An. gambiae sensu lato collected in 48 villages at varying distances to alluvial larval habitat along the River Gambia. The proportion dispersing declined exponentially with distance, and we estimated that 90% of movements were within 1.7 km. Although a ‘heavy-tailed’ distribution is considered biologically more plausible due to active dispersal by mosquitoes seeking blood meals, there was no statistical basis for choosing it over a negative exponential distribution. Using a simple random walk model with daily survival and movements previously recorded in Burkina Faso, we were able to reproduce the dispersal probabilities observed in The Gambia. Conclusions/Significance Our results provide an important quantification of the probability of An. gambiae s.l. dispersal in a rural African setting typical of many parts of the continent. However, dispersal will be landscape specific and in order to generalise to other spatial configurations of habitat and hosts it will be necessary to produce tractable models of mosquito movements for operational use. We show that simple random walk models have potential. Consequently, there is a pressing need for new empirical studies of An. gambiae survival and movements in different settings to drive this development. PMID:23874719

  20. Silencing of genes and alleles by RNAi in Anopheles gambiae.

    PubMed

    Lamacchia, Marina; Clayton, John R; Wang-Sattler, Rui; Steinmetz, Lars M; Levashina, Elena A; Blandin, Stéphanie A

    2013-01-01

    Anopheles gambiae mosquitoes are the major vectors of human malaria parasites. However, mosquitoes are not passive hosts for parasites, actively limiting their development in vivo. Our current understanding of the mosquito antiparasitic response is mostly based on the phenotypic analysis of gene knockdowns obtained by RNA interference (RNAi), through the injection or transfection of long dsRNAs in adult mosquitoes or cultured cells, respectively. Recently, RNAi has been extended to silence specifically one allele of a given gene in a heterozygous context, thus allowing to compare the contribution of different alleles to a phenotype in the same genetic background. PMID:22990777

  1. G Protein-Coupled Receptors in Anopheles gambiae

    NASA Astrophysics Data System (ADS)

    Hill, Catherine A.; Fox, A. Nicole; Pitts, R. Jason; Kent, Lauren B.; Tan, Perciliz L.; Chrystal, Mathew A.; Cravchik, Anibal; Collins, Frank H.; Robertson, Hugh M.; Zwiebel, Laurence J.

    2002-10-01

    We used bioinformatic approaches to identify a total of 276 G protein-coupled receptors (GPCRs) from the Anopheles gambiae genome. These include GPCRs that are likely to play roles in pathways affecting almost every aspect of the mosquito's life cycle. Seventy-nine candidate odorant receptors were characterized for tissue expression and, along with 76 putative gustatory receptors, for their molecular evolution relative to Drosophila melanogaster. Examples of lineage-specific gene expansions were observed as well as a single instance of unusually high sequence conservation.

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

    PubMed

    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. PMID:27365445

  3. Insecticide susceptibility of Anopheles coluzzii and Anopheles gambiae mosquitoes in Ibadan, South-West Nigeria

    PubMed Central

    Okorie, Patricia N.; Ademowo, George O.; Irving, Helen; Kelly-Hope, Louise A.; Wondji, Charles S.

    2014-01-01

    The emergence of insecticide resistance in Anopheles mosquitoes has great implications for malaria control in Nigeria. This study aimed to determine the dynamics of insecticide susceptibility levels and frequency of knockdown resistance (kdr) mutations (L1014F) in wild Anopheles coluzzii Coetzee & Wilkerson sp. n. and An. gambiae Giles ( (Diptera: Culicidae) from Ojoo and Bodija areas of Ibadan, South-West, Nigeria. Insecticide susceptibility to pyrethroids, organophosphates, carbamates and organochlorines was assessed using WHO bioassays. A subset of the mosquitoes exposed to pyrethroids and DDT was used for species and molecular form identification and kdr genotyping was determined using the Taqman real time PCR assay. The mosquitoes were resistant to pyrethroids and DDT but completely susceptible to organophosphates and carbamates. Bodja samples (n=186) comprised of An. gambiae (91.4%) and An. coluzzii (8.1%) while one An. coluzzii / An. gambiae hybrid was recorded. All mosquitoes screened in Ojoo (n=26) were An. gambiae. The 1014F kdr mutation was detected at a frequency of 24.52% 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 resistance and highlights the need to implement insecticide resistance management strategies. PMID:25417803

  4. Dosage Compensation in the African Malaria Mosquito Anopheles gambiae

    PubMed Central

    Rose, Graham; Krzywinska, Elzbieta; Kim, Jan; Revuelta, Loic; Ferretti, Luca; Krzywinski, Jaroslaw

    2016-01-01

    Dosage compensation is the fundamental process by which gene expression from the male monosomic X chromosome and from the diploid set of autosomes is equalized. Various molecular mechanisms have evolved in different organisms to achieve this task. In Drosophila, genes on the male X chromosome are upregulated to the levels of expression from the two X chromosomes in females. To test whether a similar mechanism is operating in immature stages of Anopheles mosquitoes, we analyzed global gene expression in the Anopheles gambiae fourth instar larvae and pupae using high-coverage RNA-seq data. In pupae of both sexes, the median expression ratios of X-linked to autosomal genes (X:A) were close to 1.0, and within the ranges of expression ratios between the autosomal pairs, consistent with complete compensation. Gene-by-gene comparisons of expression in males and females revealed mild female bias, likely attributable to a deficit of male-biased X-linked genes. In larvae, male to female ratios of the X chromosome expression levels were more female biased than in pupae, suggesting that compensation may not be complete. No compensation mechanism appears to operate in male germline of early pupae. Confirmation of the existence of dosage compensation in A. gambiae lays the foundation for research into the components of dosage compensation machinery in this important vector species. PMID:26782933

  5. Breakdown in the Process of Incipient Speciation in Anopheles gambiae

    PubMed Central

    Nwakanma, Davis C.; Neafsey, Daniel E.; Jawara, Musa; Adiamoh, Majidah; Lund, Emily; Rodrigues, Amabelia; Loua, Kovana M.; Konate, Lassana; Sy, Ngayo; Dia, Ibrahima; Awolola, T. Samson; Muskavitch, Marc A. T.; Conway, David J.

    2013-01-01

    Understanding genetic causes and effects of speciation in sympatric populations of sexually reproducing eukaryotes is challenging, controversial, and of practical importance for controlling rapidly evolving pests and pathogens. The major African malaria vector mosquito Anopheles gambiae sensu stricto (s.s.) is considered to contain two incipient species with strong reproductive isolation, hybrids between the M and S molecular forms being very rare. Following recent observations of higher proportions of hybrid forms at a few sites in West Africa, we conducted new surveys of 12 sites in four contiguous countries (The Gambia, Senegal, Guinea-Bissau, and Republic of Guinea). Identification and genotyping of 3499 A. gambiae s.s. revealed high frequencies of M/S hybrid forms at each site, ranging from 5 to 42%, and a large spectrum of inbreeding coefficient values from 0.11 to 0.76, spanning most of the range expected between the alternative extremes of panmixia and assortative mating. Year-round sampling over 2 years at one of the sites in The Gambia showed that M/S hybrid forms had similar relative frequencies throughout periods of marked seasonal variation in mosquito breeding and abundance. Genome-wide scans with an Affymetrix high-density single-nucleotide polymorphism (SNP) microarray enabled replicate comparisons of pools of different molecular forms, in three separate populations. These showed strong differentiation between M and S forms only in the pericentromeric region of the X chromosome that contains the molecular form-specific marker locus, with only a few other loci showing minor differences. In the X chromosome, the M/S hybrid forms were more differentiated from M than from S forms, supporting a hypothesis of asymmetric introgression and backcrossing. PMID:23335339

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

    PubMed Central

    2013-01-01

    Background Mosquitoes exhibit 24 hr rhythms in flight activity, feeding, reproduction and development. To better understand the molecular basis for these rhythms in the nocturnal malaria vector Anopheles gambiae, we have utilized microarray analysis on time-of-day specific collections of mosquitoes over 48 hr to explore the coregulation of gene expression rhythms by the circadian clock and light, and compare these with the 24 hr rhythmic gene expression in the diurnal Aedes aegypti dengue vector mosquito. Results In time courses from An. gambiae head and body collected under light:dark cycle (LD) and constant dark (DD) conditions, we applied three algorithms that detect sinusoidal patterns and an algorithm that detects spikes in expression. This revealed across four experimental conditions 393 probes newly scored as rhythmic. These genes correspond to functions such as metabolic detoxification, immunity and nutrient sensing. This includes glutathione S-transferase GSTE5, whose expression pattern and chromosomal location are shared with other genes, suggesting shared chromosomal regulation; and pulsatile expression of the gene encoding CYP6M2, a cytochrome P450 that metabolizes pyrethroid insecticides. We explored the interaction of light and the circadian clock and highlight the regulation of odorant binding proteins (OBPs), important components of the olfactory system. We reveal that OBPs have unique expression patterns as mosquitoes make the transition from LD to DD conditions. We compared rhythmic expression between An. gambiae and Ae. aegypti heads collected under LD conditions using a single cosine fitting algorithm, and report distinct similarities and differences in the temporal regulation of genes involved in tRNA priming, the vesicular-type ATPase, olfaction and vision between the two species. Conclusions These data build on our previous analyses of time-of-day specific regulation of the An. gambiae transcriptome to reveal additional rhythmic genes, an

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

  8. A new chromosomal phylogeny supports the repeated origin of vectorial capacity in malaria mosquitoes of the Anopheles gambiae complex.

    PubMed

    Kamali, Maryam; Xia, Ai; Tu, Zhijian; Sharakhov, Igor V

    2012-01-01

    Understanding phylogenetic relationships within species complexes of disease vectors is crucial for identifying genomic changes associated with the evolution of epidemiologically important traits. However, the high degree of genetic similarity among sibling species confounds the ability to determine phylogenetic relationships using molecular markers. The goal of this study was to infer the ancestral-descendant relationships among malaria vectors and nonvectors of the Anopheles gambiae species complex by analyzing breakpoints of fixed chromosomal inversions in ingroup and several outgroup species. We identified genes at breakpoints of fixed overlapping chromosomal inversions 2Ro and 2Rp of An. merus using fluorescence in situ hybridization, a whole-genome mate-paired sequencing, and clone sequencing. We also mapped breakpoints of a chromosomal inversion 2La (common to An. merus, An. gambiae, and An. arabiensis) in outgroup species using a bioinformatics approach. We demonstrated that the "standard" 2R+(p) arrangement and "inverted" 2Ro and 2La arrangements are present in outgroup species Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The data indicate that the ancestral species of the An. gambiae complex had the 2Ro, 2R+(p), and 2La chromosomal arrangements. The "inverted" 2Ro arrangement uniquely characterizes a malaria vector An. merus as the basal species in the complex. The rooted chromosomal phylogeny implies that An. merus acquired the 2Rp inversion and that its sister species An. gambiae acquired the 2R+(o) inversion from the ancestral species. The karyotype of nonvectors An. quadriannulatus A and B was derived from the karyotype of the major malaria vector An. gambiae. We conclude that the ability to effectively transmit human malaria had originated repeatedly in the complex. Our findings also suggest that saltwater tolerance originated first in An. merus and then independently in An. melas. The new chromosomal phylogeny will facilitate

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

    PubMed Central

    2011-01-01

    Background The different taxa belonging to Anopheles gambiae complex display phenotypic differences that may impact their contribution to malaria transmission. More specifically, their susceptibility to infection, resulting from a co-evolution between parasite and vector, might be different. The aim of this study was to compare the susceptibility of M and S molecular forms of Anopheles gambiae and Anopheles arabiensis to infection by Plasmodium falciparum. Methods F3 progenies of Anopheles gambiae s.l. collected in Senegal were infected, using direct membrane feeding, with P. falciparum gametocyte-containing blood sampled on volunteer patients. The presence of oocysts was determined by light microscopy after 7 days, and the presence of sporozoite by ELISA after 14 days. Mosquito species and molecular forms were identified by PCR. Results The oocyst rate was significantly higher in the molecular S form (79.07%) than in the M form (57.81%, Fisher's exact test p < 0.001) and in Anopheles arabiensis (55.38%, Fisher's exact test vs. S group p < 0.001). Mean ± s.e.m. number of oocyst was greater in the An. gambiae S form (1.72 ± 0.26) than in the An. gambiae M form (0.64 ± 0.04, p < 0.0001) and in the An. arabiensis group (0.58 ± 0.04, vs. S group, p < 0.0001). Sporozoite rate was also higher in the molecular form S (83.52%) than in form M (50.98%, Fisher's exact test p < 0.001) and Anopheles arabiensis 50.85%, Fisher's exact test vs. S group p < 0.001). Conclusion Infected in the same experimental conditions, the molecular form S of An. gambiae is more susceptible to infection by P. falciparum than the molecular form M of An. gambiae and An. arabiensis. PMID:21929746

  10. Separation of Anopheles merus from freshwater Anopheles gambiae by salinity tolerance test and morphological characters.

    PubMed

    Mosha, F W; Mutero, C M

    1982-12-01

    The separation methods for Anopheles merus from freshwater A. gambiae s.l. involving the use of salinity tolerance test, sensilla coeloconica, palpal ratio and palpal bands were evaluated for a period of one year on a total of about 340 mosquitoes. The salinity tolerance test method was found to be quite simple and reliable but unsuitable in disease transmission studies due to an interval of 2-3 days between the collection and dissection periods and also due to the fact that only a fraction of the mosquito sample is generally identified by this method. Although significantly higher proportions of sensilla coeloconica and palpal ratio were observed in A. merus as compared to freshwater A. gambiae s.l. these characters were found quite unreliable due to their overlapping between two mosquito groups. Sensilla coeloconica and palpal ratio used separately could separate respective percentages of 11.4 and 11.8 A. merus from freshwater A. gambiae s.l., while in combination they separated up to 40.9%. Percentages 4-banded palp mosquitoes accounted for about 32% in A. merus and 19% in freshwater A. gambiae s.l. All these characters also displayed some seasonal variations in the two mosquito groups. PMID:6926942

  11. Inhibition of Anopheles gambiae Odorant Receptor Function by Mosquito Repellents*

    PubMed Central

    Tsitoura, Panagiota; Koussis, Konstantinos; Iatrou, Kostas

    2015-01-01

    The identification of molecular targets of insect repellents has been a challenging task, with their effects on odorant receptors (ORs) remaining a debatable issue. Here, we describe a study on the effects of selected mosquito repellents, including the widely used repellent N,N-diethyl-meta-toluamide (DEET), on the function of specific ORs of the African malaria vector Anopheles gambiae. This study, which has been based on quantitative measurements of a Ca2+-activated photoprotein biosensor of recombinant OR function in an insect cell-based expression platform and a sequential compound addition protocol, revealed that heteromeric OR (ORx/Orco) function was susceptible to strong inhibition by all tested mosquito repellents except DEET. Moreover, our results demonstrated that the observed inhibition was due to efficient blocking of Orco (olfactory receptor coreceptor) function. This mechanism of repellent action, which is reported for the first time, is distinct from the mode of action of other characterized insect repellents including DEET. PMID:25657000

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

    PubMed Central

    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

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

    PubMed

    Tsitoura, Panagiota; Koussis, Konstantinos; Iatrou, Kostas

    2015-03-20

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

  14. Transmission potential of Rickettsia felis infection by Anopheles gambiae mosquitoes

    PubMed Central

    Dieme, Constentin; Bechah, Yassina; Socolovschi, Cristina; Audoly, Gilles; Berenger, Jean-Michel; Faye, Ousmane; Raoult, Didier; Parola, Philippe

    2015-01-01

    A growing number of recent reports have implicated Rickettsia felis as a human pathogen, paralleling the increasing detection of R. felis in arthropod hosts across the globe, primarily in fleas. Here Anopheles gambiae mosquitoes, the primary malarial vectors in sub-Saharan Africa, were fed with either blood meal infected with R. felis or infected cellular media administered in membrane feeding systems. In addition, a group of mosquitoes was fed on R. felis-infected BALB/c mice. The acquisition and persistence of R. felis in mosquitoes was demonstrated by quantitative PCR detection of the bacteria up to day 15 postinfection. R. felis was detected in mosquito feces up to day 14. Furthermore, R. felis was visualized by immunofluorescence in salivary glands, in and around the gut, and in the ovaries, although no vertical transmission was observed. R. felis was also found in the cotton used for sucrose feeding after the mosquitoes were fed infected blood. Natural bites from R. felis-infected An. gambiae were able to cause transient rickettsemias in mice, indicating that this mosquito species has the potential to be a vector of R. felis infection. This is particularly important given the recent report of high prevalence of R. felis infection in patients with “fever of unknown origin” in malaria-endemic areas. PMID:26056256

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

    SciTech Connect

    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.

  16. Chromosome end elongation by recombination in the mosquito Anopheles gambiae.

    PubMed Central

    Roth, C W; Kobeski, F; Walter, M F; Biessmann, H

    1997-01-01

    One of the functions of telomeres is to counteract the terminal nucleotide loss associated with DNA replication. While the vast majority of eukaryotic organisms maintain their chromosome ends via telomerase, an enzyme system that generates short, tandem repeats on the ends of chromosomes, other mechanisms such as the transposition of retrotransposons or recombination can also be used in some species. Chromosome end regression and extension were studied in a medically important mosquito, the malaria vector Anopheles gambiae, to determine how this dipteran insect maintains its chromosome ends. The insertion of a transgenic pUChsneo plasmid at the left end of chromosome 2 provided a unique marker for measuring the dynamics of the 2L telomere over a period of about 3 years. The terminal length was relatively uniform in the 1993 population with the chromosomes ending within the white gene sequence of the inserted transgene. Cloned terminal chromosome fragments did not end in short repeat sequences that could have been synthesized by telomerase. By late 1995, the chromosome ends had become heterogeneous: some had further shortened while other chromosomes had been elongated by regenerating part of the integrated pUChsneo plasmid. A model is presented for extension of the 2L chromosome by recombination between homologous 2L chromosome ends by using the partial plasmid duplication generated during its original integration. It is postulated that this mechanism is also important in wild-type telomere elongation. PMID:9271395

  17. Ligand-binding study of Anopheles gambiae chemosensory proteins.

    PubMed

    Iovinella, Immacolata; Bozza, Francesco; Caputo, Beniamino; Della Torre, Alessandra; Pelosi, Paolo

    2013-06-01

    Chemosensory proteins (CSPs) are a class of small proteins expressed only in arthropods and endowed with heterogeneous functions. Some of them are involved in chemical communications, others in development or other physiological roles. The numbers of CSPs in different species of insects range from 4 in Drosophila to at least 70 in locusts, whereas in other arthropods such as crustaceans and millipedes, only 2-3 very similar sequences have been reported in each species. We have expressed, in a bacterial system, 5 of the 8 CSPs predicted by the genome of the malaria mosquito Anopheles gambiae, 4 identified at the protein level (SAP1, SAP2, SAP3, and CSP3) and a fifth annotated as part of this work, obtaining the proteins with high yields and in their soluble forms. Purified CSPs have been used to study their ligand-binding properties, both using competitive binding assays and quenching of intrinsic tryptophan fluorescence, in order to get insights into their physiological functions. The agreement between the 2 sets of data supports the assumptions that the ligands, including the fluorescent reporter, bind within the core of the proteins. Their different affinities toward a set of pure chemicals suggest specific roles in chemical communication.

  18. Inactivation of Anopheles gambiae Glutathione Transferase ε2 by Epiphyllocoumarin

    PubMed Central

    Marimo, Patience; Hayeshi, Rose; Mukanganyama, Stanley

    2016-01-01

    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. PMID:26925266

  19. The “Far-West” of Anopheles gambiae Molecular Forms

    PubMed Central

    Caputo, Beniamino; Santolamazza, Federica; Vicente, José L.; Nwakanma, Davis C.; Jawara, Musa; Palsson, Katinka; Jaenson, Thomas; White, Bradley J.; Mancini, Emiliano; Petrarca, Vincenzo; Conway, David J.; Besansky, Nora J.; Pinto, João; Torre, Alessandra della

    2011-01-01

    The main Afrotropical malaria vector, Anopheles gambiae sensu stricto, is undergoing a process of sympatric ecological diversification leading to at least two incipient species (the M and S molecular forms) showing heterogeneous levels of divergence across the genome. The physically unlinked centromeric regions on all three chromosomes of these closely related taxa contain fixed nucleotide differences which have been found in nearly complete linkage disequilibrium in geographic areas of no or low M-S hybridization. Assays diagnostic for SNP and structural differences between M and S forms in the three centromeric regions were applied in samples from the western extreme of their range of sympatry, the only area where high frequencies of putative M/S hybrids have been reported. The results reveal a level of admixture not observed in the rest of the range. In particular, we found: i) heterozygous genotypes at each marker, although at frequencies lower than expected under panmixia; ii) virtually all possible genotypic combinations between markers on different chromosomes, although genetic association was nevertheless detected; iii) discordant M and S genotypes at two X-linked markers near the centromere, suggestive of introgression and inter-locus recombination. These results could be indicative either of a secondary contact zone between M and S, or of the maintenance of ancestral polymorphisms. This issue and the perspectives opened by these results in the study of the M and S incipient speciation process are discussed. PMID:21347223

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

    PubMed Central

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

    PubMed Central

    Hume, Jennifer CC; Tunnicliff, Mark; Ranford-Cartwright, Lisa C; Day, Karen P

    2007-01-01

    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. PMID:17958900

  2. Identification and Characterization of Two Novel RNA Viruses from Anopheles gambiae Species Complex Mosquitoes

    PubMed Central

    Carissimo, Guillaume; Eiglmeier, Karin; Reveillaud, Julie; Holm, Inge; Diallo, Mawlouth; Diallo, Diawo; Vantaux, Amélie; Kim, Saorin; Ménard, Didier; Siv, Sovannaroth; Belda, Eugeni; Bischoff, Emmanuel; Antoniewski, Christophe; Vernick, Kenneth D.

    2016-01-01

    Mosquitoes of the Anopheles gambiae complex display strong preference for human bloodmeals and are major malaria vectors in Africa. However, their interaction with viruses or role in arbovirus transmission during epidemics has been little examined, with the exception of O’nyong-nyong virus, closely related to Chikungunya virus. Deep-sequencing has revealed different RNA viruses in natural insect viromes, but none have been previously described in the Anopheles gambiae species complex. Here, we describe two novel insect RNA viruses, a Dicistrovirus and a Cypovirus, found in laboratory colonies of An. gambiae taxa using small-RNA deep sequencing. Sequence analysis was done with Metavisitor, an open-source bioinformatic pipeline for virus discovery and de novo genome assembly. Wild-collected Anopheles from Senegal and Cambodia were positive for the Dicistrovirus and Cypovirus, displaying high sequence identity to the laboratory-derived virus. Thus, the Dicistrovirus (Anopheles C virus, AnCV) and Cypovirus (Anopheles Cypovirus, AnCPV) are components of the natural virome of at least some anopheline species. Their possible influence on mosquito immunity or transmission of other pathogens is unknown. These natural viruses could be developed as models for the study of Anopheles-RNA virus interactions in low security laboratory settings, in an analogous manner to the use of rodent malaria parasites for studies of mosquito anti-parasite immunity. PMID:27138938

  3. A High-Affinity Adenosine Kinase from Anopheles Gambiae

    SciTech Connect

    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.

  4. Seasonal climate effects anemotaxis in newly emerged adult anopheles gambiae giles in Mali, West Africa

    Technology Transfer Automated Retrieval System (TEKTRAN)

    The direction and magnitude of movement of the malaria vector Anopheles gambiae Giles has been of great interest to medical entomologists for over 70 years. This direction of movement is likely to be affected by many factors, from environmental conditions and stage of life history of the mosquito to...

  5. Equivalent susceptibility of Anopheles gambiae M and S molecular forms and Anopheles arabiensis to Plasmodium falciparum infection in Burkina Faso

    PubMed Central

    2013-01-01

    Background The Anopheles gambiae sensu lato (s.l.) species complex in Burkina Faso consists of Anopheles arabiensis, and molecular forms M and S of Anopheles gambiae sensu stricto (s.s.). Previous studies comparing the M and S forms for level of infection with Plasmodium falciparum have yielded conflicting results. Methods Mosquito larvae were sampled from natural pools, reared to adulthood under controlled conditions, and challenged with natural P. falciparum by experimental feeding with blood from gametocyte carriers. Oocyst infection prevalence and intensity was determined one week after infection. DNA from carcasses was genotyped to identify species and molecular form. Results In total, 7,400 adult mosquitoes grown from wild-caught larvae were challenged with gametocytes in 29 experimental infections spanning four transmission seasons. The overall infection prevalence averaged 40.7% for A. gambiae M form, 41.4% for A. gambiae S form, and 40.1% for A. arabiensis. There was no significant difference in infection prevalence or intensity between the three population groups. Notably, infection experiments in which the population groups were challenged in parallel on the same infective blood displayed less infection difference between population groups, while infections with less balanced composition of population groups had lower statistical power and displayed apparent differences that fluctuated more often from the null average. Conclusion The study clearly establishes that, at the study site in Burkina Faso, there is no difference in genetic susceptibility to P. falciparum infection between three sympatric population groups of the A. gambiae s.l. complex. Feeding the mosquito groups on the same infective blood meal greatly increases statistical power. Conversely, comparison of the different mosquito groups between, rather than within, infections yields larger apparent difference between mosquito groups, resulting from lower statistical power and greater noise

  6. Evidence for a Lectin Specific for Sulfated Glycans in the Salivary Gland of the Malaria Vector, Anopheles gambiae

    PubMed Central

    Francischetti, Ivo M. B.; Ma, Dongying; Andersen, John F.; Ribeiro, José M. C.

    2014-01-01

    Salivary gland homogenate (SGH) from the female mosquitoes Anopheles gambiae, An. stephensi, An. freeborni, An. dirus and An. albimanus were found to exhibit hemagglutinating (lectin) activity. Lectin activity was not found for male An. gambiae, or female Ae aegypti, Culex quinquefasciatus, Phlebotomus duboscqi, and Lutzomyia longipalpis. With respect to species-specificity, An. gambiae SGH agglutinates red blood cells (RBC) from humans, horse, sheep, goat, pig, and cow; it is less active for rats RBC, and not detectable for guinea-pigs or chicken RBC. Notably, lectin activity was inhibited by low concentrations of dextran sulfate 50–500 K, fucoidan, heparin, laminin, heparin sulfate proteoglycan, sialyl-containing glycans (e.g. 3′-sialyl Lewis X, and 6′-sialyl lactose), and gangliosides (e.g. GM3, GD1, GD1b, GTB1, GM1, GQ1B), but not by simple sugars. These results imply that molecule(s) in the salivary gland target sulfated glycans. SGH from An. gambiae was also found to promote agglutination of HL-60 cells which are rich in sialyl Lewis X, a glycan that decorates PSGL-1, the neutrophils receptor that interacts with endothelial cell P-selectin. Accordingly, SGH interferes with HL-60 cells adhesion to immobilized P-selectin. Because An. gambiae SGH expresses galectins, one member of this family (herein named Agalectin) was expressed in E. coli. Recombinant Agalectin behaves as a non-covalent homodimer. It does not display lectin activity, and does not interact with 500 candidates tested in a Glycan microarray. Gel-filtration chromatography of the SGH of An. gambiae identified a fraction with hemagglutinating activity, which was analyzed by 1D PAGE followed by in-gel tryptic digestion, and nano-LC MS/MS. This approach identified several genes which emerge as candidates for a lectin targeting sulfated glycans, the first with this selectivity to be reported in the SGH of a blood-sucking arthropod. The role of salivary molecules (sialogenins) with lectin

  7. Evidence for a lectin specific for sulfated glycans in the salivary gland of the malaria vector, Anopheles gambiae.

    PubMed

    Francischetti, Ivo M B; Ma, Dongying; Andersen, John F; Ribeiro, José M C

    2014-01-01

    Salivary gland homogenate (SGH) from the female mosquitoes Anopheles gambiae, An. stephensi, An. freeborni, An. dirus and An. albimanus were found to exhibit hemagglutinating (lectin) activity. Lectin activity was not found for male An. gambiae, or female Ae aegypti, Culex quinquefasciatus, Phlebotomus duboscqi, and Lutzomyia longipalpis. With respect to species-specificity, An. gambiae SGH agglutinates red blood cells (RBC) from humans, horse, sheep, goat, pig, and cow; it is less active for rats RBC, and not detectable for guinea-pigs or chicken RBC. Notably, lectin activity was inhibited by low concentrations of dextran sulfate 50-500 K, fucoidan, heparin, laminin, heparin sulfate proteoglycan, sialyl-containing glycans (e.g. 3'-sialyl Lewis X, and 6'-sialyl lactose), and gangliosides (e.g. GM3, GD1, GD1b, GTB1, GM1, GQ1B), but not by simple sugars. These results imply that molecule(s) in the salivary gland target sulfated glycans. SGH from An. gambiae was also found to promote agglutination of HL-60 cells which are rich in sialyl Lewis X, a glycan that decorates PSGL-1, the neutrophils receptor that interacts with endothelial cell P-selectin. Accordingly, SGH interferes with HL-60 cells adhesion to immobilized P-selectin. Because An. gambiae SGH expresses galectins, one member of this family (herein named Agalectin) was expressed in E. coli. Recombinant Agalectin behaves as a non-covalent homodimer. It does not display lectin activity, and does not interact with 500 candidates tested in a Glycan microarray. Gel-filtration chromatography of the SGH of An. gambiae identified a fraction with hemagglutinating activity, which was analyzed by 1D PAGE followed by in-gel tryptic digestion, and nano-LC MS/MS. This approach identified several genes which emerge as candidates for a lectin targeting sulfated glycans, the first with this selectivity to be reported in the SGH of a blood-sucking arthropod. The role of salivary molecules (sialogenins) with lectin activity is

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

    PubMed Central

    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

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

    PubMed

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

    2016-01-01

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

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

    PubMed

    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. PMID:27322614

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

    PubMed Central

    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 the two major and widespread malaria vectors in sub-Saharan Africa. Several studies have examined the ecology and population dynamics of these vectors. Ecological conditions along the Kilombero valley in Tanzania influence the distribution and population density of these two vector species. It remains unclear whether the ecological diversity within the Kilombero valley has affected the population structure of An. gambiae s.l. populations. The goal of this study was to characterise the genetic structure of sympatric An. gambiae s.s and An. arabiensis populations along the Kilombero valley. Methodology Mosquitoes were collected from seven locations in Tanzania: six from the Kilombero valley and one outside the valley (~700 km away) as an out-group. To archive a genome-wide coverage, 13 microsatellite markers from chromosomes X, 2 and 3 were used. Results High levels of genetic differentiation among An. arabiensis populations was observed, as opposed to An. gambiae s.s., which was genetically undifferentiated across the 6,650 km2 of the Kilombero valley landscape. It appears that genetic differentiation is not attributed to physical barriers or distance, but possibly by ecological diversification within the Kilombero valley. Genetic divergence among An. arabiensis populations (FST = 0.066) was higher than that of the well-known M and S forms of An. gambiae s. s. in West and Central Africa (FST = 0.035), suggesting that these populations are maintained by some level of reproductive isolation. Conclusion It was hypothesized that ecological diversification across the valley may be a driving force for observed An. arabiensis genetic divergence. The impact of the observed An

  12. Anopheles gambiae Antiviral Immune Response to Systemic O'nyong-nyong Infection

    PubMed Central

    Waldock, Joanna; Olson, Kenneth E.; Christophides, George K.

    2012-01-01

    Background Mosquito-borne viral diseases cause significant burden in much of the developing world. Although host-virus interactions have been studied extensively in the vertebrate host, little is known about mosquito responses to viral infection. In contrast to mosquitoes of the Aedes and Culex genera, Anopheles gambiae, the principal vector of human malaria, naturally transmits very few arboviruses, the most important of which is O'nyong-nyong virus (ONNV). Here we have investigated the A. gambiae immune response to systemic ONNV infection using forward and reverse genetic approaches. Methodology/Principal Findings We have used DNA microarrays to profile the transcriptional response of A. gambiae inoculated with ONNV and investigate the antiviral function of candidate genes through RNAi gene silencing assays. Our results demonstrate that A. gambiae responses to systemic viral infection involve genes covering all aspects of innate immunity including pathogen recognition, modulation of immune signalling, complement-mediated lysis/opsonisation and other immune effector mechanisms. Patterns of transcriptional regulation and co-infections of A. gambiae with ONNV and the rodent malaria parasite Plasmodium berghei suggest that hemolymph immune responses to viral infection are diverted away from melanisation. We show that four viral responsive genes encoding two putative recognition receptors, a galectin and an MD2-like receptor, and two effector lysozymes, function in limiting viral load. Conclusions/Significance This study is the first step in elucidating the antiviral mechanisms of A. gambiae mosquitoes, and has revealed interesting differences between A. gambiae and other invertebrates. Our data suggest that mechanisms employed by A. gambiae are distinct from described invertebrate antiviral immunity to date, and involve the complement-like branch of the humoral immune response, supressing the melanisation response that is prominent in anti-parasitic immunity. The

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

    PubMed

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

    2012-07-01

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

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

    PubMed

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

    1996-06-01

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

  15. Cloning and analysis of a cecropin gene from the malaria vector mosquito, Anopheles gambiae.

    PubMed

    Vizioli, J; Bulet, P; Charlet, M; Lowenberger, C; Blass, C; Müller, H M; Dimopoulos, G; Hoffmann, J; Kafatos, F C; Richman, A

    2000-02-01

    Parasites of the genus Plasmodium are transmitted to mammalian hosts by anopheline mosquitoes. Within the insect vector, parasite growth and development are potentially limited by antimicrobial defence molecules. Here, we describe the isolation of cDNA and genomic clones encoding a cecropin antibacterial peptide from the malaria vector mosquito Anopheles gambiae. The locus was mapped to polytene division 1C of the X chromosome. Cecropin RNA was induced by infection with bacteria and Plasmodium. RNA levels varied in different body parts of the adult mosquito. During development, cecropin expression was limited to the early pupal stage. The peptide was purified from both adult mosquitoes and cell culture supernatants. Anopheles gambiae synthetic cecropins displayed activity against Gram-negative and Gram-positive bacteria, filamentous fungi and yeasts. PMID:10672074

  16. Beta-integrin of Anopheles gambiae: mRNA cloning and analysis of structure and expression.

    PubMed

    Mahairaki, V; Lycett, G; Blass, C; Louis, C

    2001-06-01

    We have isolated an mRNA encoding a beta integrin subunit of the malaria mosquito Anopheles gambiae. Our analysis predicts a protein that is very similar to betaPS, the fruitfly orthologue. The gene is expressed during all developmental stages and it is found in all body parts, including the midgut. Finally, the expression of the gene does not seem to be modulated during blood meals, except for a substantial increase 48 h posthaematophagy, when digestion is nearly complete. PMID:11437913

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

    PubMed

    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.

  18. Assessing the Drosophila melanogaster and Anopheles gambiae Genome Annotations Using Genome-Wide Sequence Comparisons

    PubMed Central

    Jaillon, Olivier; Dossat, Carole; Eckenberg, Ralph; Eiglmeier, Karin; Segurens, Béatrice; Aury, Jean-Marc; Roth, Charles W.; Scarpelli, Claude; Brey, Paul T.; Weissenbach, Jean; Wincker, Patrick

    2003-01-01

    We performed genome-wide sequence comparisons at the protein coding level between the genome sequences of Drosophila melanogaster and Anopheles gambiae. Such comparisons detect evolutionarily conserved regions (ecores) that can be used for a qualitative and quantitative evaluation of the available annotations of both genomes. They also provide novel candidate features for annotation. The percentage of ecores mapping outside annotations in the A. gambiae genome is about fourfold higher than in D. melanogaster. The A. gambiae genome assembly also contains a high proportion of duplicated ecores, possibly resulting from artefactual sequence duplications in the genome assembly. The occurrence of 4063 ecores in the D. melanogaster genome outside annotations suggests that some genes are not yet or only partially annotated. The present work illustrates the power of comparative genomics approaches towards an exhaustive and accurate establishment of gene models and gene catalogues in insect genomes. PMID:12840038

  19. Distribution and spread of pyrethroid and DDT resistance among the Anopheles gambiae complex in Tanzania.

    PubMed

    Kabula, B; Tungu, P; Malima, R; Rowland, M; Minja, J; Wililo, R; Ramsan, M; McElroy, P D; Kafuko, J; Kulkarni, M; Protopopoff, N; Magesa, S; Mosha, F; Kisinza, W

    2014-09-01

    The development of insecticide resistance is a threat to the control of malaria in Africa. We report the findings of a national survey carried out in Tanzania in 2011 to monitor the susceptibility of malaria vectors to pyrethroid, organophosphate, carbamate and DDT insecticides, and compare these findings with those identified in 2004 and 2010. Standard World Health Organization (WHO) methods were used to detect knock-down and mortality rates in wild female Anopheles gambiae s.l. (Diptera: Culicidae) collected from 14 sentinel districts. Diagnostic doses of the pyrethroids deltamethrin, lambdacyhalothrin and permethrin, the carbamate propoxur, the organophosphate fenitrothion and the organochlorine DDT were used. Anopheles gambiae s.l. was resistant to permethrin in Muleba, where a mortality rate of 11% [95% confidence interval (CI) 6-19%] was recorded, Muheza (mortality rate of 75%, 95% CI 66-83%), Moshi and Arumeru (mortality rates of 74% in both). Similarly, resistance was reported to lambdacyhalothrin in Muleba, Muheza, Moshi and Arumeru (mortality rates of 31-82%), and to deltamethrin in Muleba, Moshi and Muheza (mortality rates of 28-75%). Resistance to DDT was reported in Muleba. No resistance to the carbamate propoxur or the organophosphate fenitrothion was observed. Anopheles gambiae s.l. is becoming resistant to pyrethoids and DDT in several parts of Tanzania. This has coincided with the scaling up of vector control measures. Resistance may impair the effectiveness of these interventions and therefore demands close monitoring and the adoption of a resistance management strategy.

  20. Variation in metabolic rate of Anopheles gambiae and A. arabiensis in a Sahelian village.

    PubMed

    Huestis, Diana L; Yaro, Alpha S; Traoré, Adama I; Adamou, Abdoulaye; Kassogué, Yaya; Diallo, Moussa; Timbiné, Seydou; Dao, Adama; Lehmann, Tovi

    2011-07-15

    In the Sahel, the Anopheles gambiae complex consists of Anopheles arabiensis and the M and S molecular forms of A. gambiae sensu stricto. However, the composition of these malaria vectors varies spatially and temporally throughout the region and is thought to be linked to environmental factors such as rainfall, larval site characteristics and duration of the dry season. To examine possible physiological divergence between these taxa, we measured metabolic rates of mosquitoes during the wet season in a Sahelian village in Mali. To our knowledge, this study provides the first measurements of metabolic rates of A. gambiae and A. arabiensis in the field. The mean metabolic rate of A. arabiensis was higher than that of M-form A. gambiae when accounting for the effects of female gonotrophic status, temperature and flight activity. However, after accounting for their difference in body size, no significant difference in metabolic rate was found between these two species (whilst all other factors were found to be significant). Thus, body size may be a key character that has diverged in response to ecological differences between these two species. Alternatively, these species may display additional differences in metabolic rate only during the dry season. Overall, our results indicate that changes in behavior and feeding activity provide an effective mechanism for mosquitoes to reduce their metabolic rate, and provide insight into the possible strategies employed by aestivating individuals during the dry season. We hypothesize that female mosquitoes switch to sugar feeding while in dormancy because of elevated metabolism associated with blood digestion.

  1. Filling the Gap 115 Years after Ronald Ross: The Distribution of the Anopheles coluzzii and Anopheles gambiae s.s from Freetown and Monrovia, West Africa

    PubMed Central

    de Souza, Dziedzom K.; Koudou, Benjamin G.; Bolay, Fatorma K.; Boakye, Daniel A.; Bockarie, Moses J.

    2013-01-01

    It was in Freetown, Sierra Leone, that the malaria mosquito Anopheles coastalis, now known as Anopheles gambiae, was first discovered as the vector of malaria, in 1899. That discovery led to a pioneering vector research in Sierra Leone and neighbouring Liberia, where mosquito species were extensively characterized. Unfortunately, the decade long civil conflicts of the 1990s, in both countries, resulted in a stagnation of the once vibrant research on disease vectors. This paper attempts to fill in some of the gaps on what is now known of the distribution of the sibling species of the An. gambiae complex, and especially the An. coluzzii and An. gambiae s.s, formerly known as the An. gambiae molecular M and S forms respectively, in the cities of Freetown and Monrovia. PMID:23741429

  2. A test of the chromosomal theory of ecotypic speciation in Anopheles gambiae.

    PubMed

    Manoukis, Nicholas C; Powell, Jeffrey R; Touré, Mahamoudou B; Sacko, Adama; Edillo, Frances E; Coulibaly, Mamadou B; Traoré, Sekou F; Taylor, Charles E; Besansky, Nora J

    2008-02-26

    The role of chromosomal inversions in speciation has long been of interest to evolutionists. Recent quantitative modeling has stimulated reconsideration of previous conceptual models for chromosomal speciation. Anopheles gambiae, the most important vector of human malaria, carries abundant chromosomal inversion polymorphism nonrandomly associated with ecotypes that mate assortatively. Here, we consider the potential role of paracentric inversions in promoting speciation in A. gambiae via "ecotypification," a term that refers to differentiation arising from local adaptation. In particular, we focus on the Bamako form, an ecotype characterized by low inversion polymorphism and fixation of an inversion, 2Rj, that is very rare or absent in all other forms of A. gambiae. The Bamako form has a restricted distribution by the upper Niger River and its tributaries that is associated with a distinctive type of larval habitat, laterite rock pools, hypothesized to be its optimal breeding site. We first present computer simulations to investigate whether the population dynamics of A. gambiae are consistent with chromosomal speciation by ecotypification. The models are parameterized using field observations on the various forms of A. gambiae that exist in Mali, West Africa. We then report on the distribution of larvae of this species collected from rock pools and more characteristic breeding sites nearby. Both the simulations and field observations support the thesis that speciation by ecotypification is occurring, or has occurred, prompting consideration of Bamako as an independent species. PMID:18287019

  3. A test of the chromosomal theory of ecotypic speciation in Anopheles gambiae.

    PubMed

    Manoukis, Nicholas C; Powell, Jeffrey R; Touré, Mahamoudou B; Sacko, Adama; Edillo, Frances E; Coulibaly, Mamadou B; Traoré, Sekou F; Taylor, Charles E; Besansky, Nora J

    2008-02-26

    The role of chromosomal inversions in speciation has long been of interest to evolutionists. Recent quantitative modeling has stimulated reconsideration of previous conceptual models for chromosomal speciation. Anopheles gambiae, the most important vector of human malaria, carries abundant chromosomal inversion polymorphism nonrandomly associated with ecotypes that mate assortatively. Here, we consider the potential role of paracentric inversions in promoting speciation in A. gambiae via "ecotypification," a term that refers to differentiation arising from local adaptation. In particular, we focus on the Bamako form, an ecotype characterized by low inversion polymorphism and fixation of an inversion, 2Rj, that is very rare or absent in all other forms of A. gambiae. The Bamako form has a restricted distribution by the upper Niger River and its tributaries that is associated with a distinctive type of larval habitat, laterite rock pools, hypothesized to be its optimal breeding site. We first present computer simulations to investigate whether the population dynamics of A. gambiae are consistent with chromosomal speciation by ecotypification. The models are parameterized using field observations on the various forms of A. gambiae that exist in Mali, West Africa. We then report on the distribution of larvae of this species collected from rock pools and more characteristic breeding sites nearby. Both the simulations and field observations support the thesis that speciation by ecotypification is occurring, or has occurred, prompting consideration of Bamako as an independent species.

  4. Multimodal Pyrethroid Resistance in Malaria Vectors, Anopheles gambiae s.s., Anopheles arabiensis, and Anopheles funestus s.s. in Western Kenya

    PubMed Central

    Kawada, Hitoshi; Dida, Gabriel O.; Ohashi, Kazunori; Komagata, Osamu; Kasai, Shinji; Tomita, Takashi; Sonye, George; Maekawa, Yoshihide; Mwatele, Cassian; Njenga, Sammy M.; Mwandawiro, Charles; Minakawa, Noboru; Takagi, Masahiro

    2011-01-01

    Anopheles gambiae s.s., Anopheles arabiensis, and Anopheles funestus s.s. are the most important species for malaria transmission. Pyrethroid resistance of these vector mosquitoes is one of the main obstacles against effective vector control. The objective of the present study was to monitor the pyrethroid susceptibility in the 3 major malaria vectors in a highly malaria endemic area in western Kenya and to elucidate the mechanisms of pyrethroid resistance in these species. Gembe East and West, Mbita Division, and 4 main western islands in the Suba district of the Nyanza province in western Kenya were used as the study area. Larval and adult collection and bioassay were conducted, as well as the detection of point mutation in the voltage-gated sodium channel (1014L) by using direct DNA sequencing. A high level of pyrethroid resistance caused by the high frequency of point mutations (L1014S) was detected in An. gambiae s.s. In contrast, P450-related pyrethroid resistance seemed to be widespread in both An. arabiensis and An. funestus s.s. Not a single L1014S mutation was detected in these 2 species. A lack of cross-resistance between DDT and permethrin was also found in An. arabiensis and An. funestus s.s., while An. gambiae s.s. was resistant to both insecticides. It is noteworthy that the above species in the same area are found to be resistant to pyrethroids by their unique resistance mechanisms. Furthermore, it is interesting that 2 different resistance mechanisms have developed in the 2 sibling species in the same area individually. The cross resistance between permethrin and DDT in An. gambiae s.s. may be attributed to the high frequency of kdr mutation, which might be selected by the frequent exposure to ITNs. Similarly, the metabolic pyrethroid resistance in An. arabiensis and An. funestus s.s. is thought to develop without strong selection by DDT. PMID:21853038

  5. Mass spectrometry identification of age-associated proteins from the malaria mosquitoes Anopheles gambiae s.s. and Anopheles stephensi.

    PubMed

    Sikulu, Maggy T; Monkman, James; Dave, Keyur A; Hastie, Marcus L; Dale, Patricia E; Kitching, Roger L; Killeen, Gerry F; Kay, Brian H; Gorman, Jeffry J; Hugo, Leon E

    2015-09-01

    This study investigated proteomic changes occurring in Anopheles gambiae and Anopheles stephensi during adult mosquito aging. These changes were evaluated using two-dimensional difference gel electrophoresis (2D-DIGE) and the identities of aging related proteins were determined using capillary high-pressure liquid chromatography (capHPLC) coupled with a linear ion-trap (LTQ)-Orbitrap XL hybrid mass spectrometry (MS). Here, we have described the techniques used to determine age associated proteomic changes occurring in heads and thoraces across three age groups; 1, 9 and 17 d old A. gambiae and 4 age groups; 1, 9, 17 and 34 d old A. stephensi. We have provided normalised spot volume raw data for all protein spots that were visible on 2D-DIGE images for both species and processed Orbitrap mass spectrometry data. For public access, mass spectrometry raw data are available via ProteomeXchange with identifier PXD002153. A detailed description of this study has been described elsewhere [1].

  6. Mass spectrometry identification of age-associated proteins from the malaria mosquitoes Anopheles gambiae s.s. and Anopheles stephensi

    PubMed Central

    Sikulu, Maggy T.; Monkman, James; Dave, Keyur A.; Hastie, Marcus L.; Dale, Patricia E.; Kitching, Roger L.; Killeen, Gerry F.; Kay, Brian H.; Gorman, Jeffry J.; Hugo, Leon E.

    2015-01-01

    This study investigated proteomic changes occurring in Anopheles gambiae and Anopheles stephensi during adult mosquito aging. These changes were evaluated using two-dimensional difference gel electrophoresis (2D-DIGE) and the identities of aging related proteins were determined using capillary high-pressure liquid chromatography (capHPLC) coupled with a linear ion-trap (LTQ)-Orbitrap XL hybrid mass spectrometry (MS). Here, we have described the techniques used to determine age associated proteomic changes occurring in heads and thoraces across three age groups; 1, 9 and 17 d old A. gambiae and 4 age groups; 1, 9, 17 and 34 d old A. stephensi. We have provided normalised spot volume raw data for all protein spots that were visible on 2D-DIGE images for both species and processed Orbitrap mass spectrometry data. For public access, mass spectrometry raw data are available via ProteomeXchange with identifier PXD002153. A detailed description of this study has been described elsewhere [1]. PMID:26306320

  7. Molasses as a source of carbon dioxide for attracting the malaria mosquitoes Anopheles gambiae and Anopheles funestus

    PubMed Central

    2014-01-01

    Background Most odour baits for haematophagous arthropods contain carbon dioxide (CO2). The CO2 is sourced artificially from the fermentation of refined sugar (sucrose), dry ice, pressurized gas cylinders or propane. These sources of CO2 are neither cost-effective nor sustainable for use in remote areas of sub-Saharan Africa. In this study, molasses was evaluated as a potential substrate for producing CO2 used as bait for malaria mosquitoes. Methods The attraction of laboratory-reared and wild Anopheles gambiae complex mosquitoes to CO2 generated from yeast-fermentation of molasses was assessed under semi-field and field conditions in western Kenya. In the field, responses of wild Anopheles funestus were also assessed. Attraction of the mosquitoes to a synthetic mosquito attractant, Mbita blend (comprising ammonia, L-lactic acid, tetradecanoic acid and 3-methyl-1-butanol) when augmented with CO2 generated from yeast fermentation of either molasses or sucrose was also investigated. Results In semi-field, the release rate of CO2 and proportion of An. gambiae mosquitoes attracted increased in tandem with an increase in the quantity of yeast-fermented molasses up to an optimal ratio of molasses and dry yeast. More An. gambiae mosquitoes were attracted to a combination of the Mbita blend plus CO2 produced from fermenting molasses than the Mbita blend plus CO2 from yeast-fermented sucrose. In the field, significantly more female An. gambiae sensu lato mosquitoes were attracted to the Mbita blend augmented with CO2 produced by fermenting 500 g of molasses compared to 250 g of sucrose or 250 g of molasses. Similarly, significantly more An. funestus, Culex and other anopheline mosquito species were attracted to the Mbita blend augmented with CO2 produced from fermenting molasses than the Mbita blend with CO2 produced from sucrose. Augmenting the Mbita blend with CO2 produced from molasses was associated with high catches of blood-fed An. gambiae s.l. and An. funestus

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

    PubMed

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

    2011-08-16

    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

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

    PubMed Central

    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

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

    PubMed

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

    2005-02-01

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

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

    NASA Astrophysics Data System (ADS)

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

    2007-03-01

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

  12. Selective Introgression of Paracentric Inversions between Two Sibling Species of the Anopheles Gambiae Complex

    PubMed Central

    Torre, A. D.; Merzagora, L.; Powell, J. R.; Coluzzi, M.

    1997-01-01

    The Anopheles gambiae complex includes the major vectors of malaria in sub-Saharan Africa where >80% of all world-wide cases occur. These mosquitoes are characterized by chromosomal inversions associated to the speciation process and to intraspecific ecological and behavioral flexibility. It has been postulated that introgressive hybridization has selectively transferred inversions on the second chromosome between A. gambiae and A. arabiensis, the two most important vectors of malaria. Here we directly test this hypothesis with laboratory experiments in which hybrid populations were established and the fate of chromosomal inversions were followed. Consistent with the hypothesis, ``foreign'' X chromosomes were eliminated within two generations, while some ``foreign'' second chromosomes persisted for the duration of the experiments and, judging from the excess of heterozygotes, established stable heterotic polymorphisms. Only those second chromosome inversions found naturally in the species could be introgressed. PMID:9136013

  13. Serosal cuticle formation and distinct degrees of desiccation resistance in embryos of the mosquito vectors Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus.

    PubMed

    Vargas, Helena Carolina Martins; Farnesi, Luana Cristina; Martins, Ademir Jesus; Valle, Denise; Rezende, Gustavo Lazzaro

    2014-03-01

    Given their medical importance, mosquitoes have been studied as vectors of parasites since the late 1800's. However, there are still many gaps concerning some aspects of their biology, such as embryogenesis. The embryonic desiccation resistance (EDR), already described in Aedes and Anopheles gambiae mosquitoes, is a peculiar trait. Freshly laid eggs are susceptible to water loss, a condition that can impair their viability. EDR is acquired during embryogenesis through the formation of the serosal cuticle (SC), protecting eggs from desiccation. Nevertheless, conservation of both traits (SC presence and EDR acquisition) throughout mosquito evolution is unknown. Comparative physiological studies with mosquito embryos from different genera, exhibiting distinct evolutionary histories and habits is a feasible approach. In this sense, the process of EDR acquisition of Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus at 25°C was evaluated. Completion of embryogenesis occurs in Ae. aegypti, An. aquasalis and Cx. quinquefasciatus at, respectively 77.4, 51.3 and 34.3hours after egg laying, Cx. quinquefasciatus embryonic development taking less than half the time of Ae. aegypti. In all cases, EDR is acquired in correlation with SC formation. For both Ae. aegypti and An. aquasalis, EDR and SC appear at 21% of total embryonic development, corresponding to the morphological stage of complete germ band elongation/beginning of germ band retraction. Although phylogenetically closer to Ae. aegypti than to An. aquasalis, Cx. quinquefasciatus acquires both EDR and serosal cuticle later, with 35% of total development, when the embryo already progresses to the middle of germ band retraction. EDR confers distinct egg viability in these species. While Ae. aegypti eggs demonstrated high viability when left up to 72hours in a dry environment, those of An. aquasalis and Cx. quinquefasciatus supported these conditions for only 24 and 5hours, respectively. Our data suggest that

  14. Implications of low-density microfilariae carriers in Anopheles transmission areas: molecular forms of Anopheles gambiae and Anopheles funestus populations in perspective

    PubMed Central

    2014-01-01

    Background Previous studies have shown a general reduction in annual transmission potential (ATP) of Anopheles species after mass drug administration (MDA) in lymphatic filariasis endemic communities. Whereas results obtained from a monitoring programme after three years of MDA revealed a decrease in ATP of Anopheles funestus this was not the same for An. gambiae s.s. in Ghana. In this study, the ability of these vectors in transmitting Wuchereria bancrofti in nine lymphatic filariasis endemic communities in Gomoa District of Ghana after four rounds of MDA with ivermectin and albendazole was investigated. Methods After mass screening of inhabitants in these communities, twelve consenting volunteers with different intensities of microfilariae (mf) slept under partly opened mosquito nets as sources of mf blood meal. Hourly collection of mosquitoes and finger-pricked blood were taken from 21.00 to 06.00 hours the following day. For each hour, half of the mosquitoes collected were immediately killed and dissected for mf. The remaining half were maintained up to 13 days for parasite maturation. Parasitaemia and infection rates in the mosquitoes were determined by microscopy. The mosquitoes were identified by microscopy and molecular techniques. Results A total of 1,083 participants were screened and the overall parasite prevalence was 1.6% with mf intensities ranging from 0 to 59 per 100 μl and geometric mean intensity of 1.1 mf per ml of blood. Of the 564 mosquitoes collected, 350 (62.1%) were Anopheles spp., from which 310 (88.6%) were An. funestus and 32 (9.1%) An. gambiae. Six anopheline mosquitoes (1.7%) were found infected with L1, but no larva was observed in any of the mosquitoes maintained up to 13 days. Molecular studies showed all An. gambiae s.l. to be An. gambiae s.s., of which 21 (70%) were of the M molecular form. Conclusion At low-level parasitaemia after 4 rounds of MDA, there was no recovery of infective stage larvae of W. bancrofti in An

  15. Variation in metabolic rate of Anopheles gambiae and A. arabiensis in a Sahelian village

    PubMed Central

    Huestis, Diana L.; Yaro, Alpha S.; Traoré, Adama I.; Adamou, Abdoulaye; Kassogué, Yaya; Diallo, Moussa; Timbiné, Seydou; Dao, Adama; Lehmann, Tovi

    2011-01-01

    SUMMARY In the Sahel, the Anopheles gambiae complex consists of Anopheles arabiensis and the M and S molecular forms of A. gambiae sensu stricto. However, the composition of these malaria vectors varies spatially and temporally throughout the region and is thought to be linked to environmental factors such as rainfall, larval site characteristics and duration of the dry season. To examine possible physiological divergence between these taxa, we measured metabolic rates of mosquitoes during the wet season in a Sahelian village in Mali. To our knowledge, this study provides the first measurements of metabolic rates of A. gambiae and A. arabiensis in the field. The mean metabolic rate of A. arabiensis was higher than that of M-form A. gambiae when accounting for the effects of female gonotrophic status, temperature and flight activity. However, after accounting for their difference in body size, no significant difference in metabolic rate was found between these two species (whilst all other factors were found to be significant). Thus, body size may be a key character that has diverged in response to ecological differences between these two species. Alternatively, these species may display additional differences in metabolic rate only during the dry season. Overall, our results indicate that changes in behavior and feeding activity provide an effective mechanism for mosquitoes to reduce their metabolic rate, and provide insight into the possible strategies employed by aestivating individuals during the dry season. We hypothesize that female mosquitoes switch to sugar feeding while in dormancy because of elevated metabolism associated with blood digestion. PMID:21697426

  16. Evaluating preservation methods for identifying Anopheles gambiae s.s. and Anopheles arabiensis complex mosquitoes species using near infra-red spectroscopy

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Near infra-red spectroscopy (NIRS) has been successfully used on fresh and RNAlater® preserved Anopheles gambiae complex to identify sibling species and age. No preservation methods other than RNAlater® have been tested to preserve mosquitoes for species identification using NIRS. However, RNAlater®...

  17. Bacterial associations reveal spatial population dynamics in Anopheles gambiae mosquitoes

    PubMed Central

    Buck, Moritz; Nilsson, Louise K. J.; Brunius, Carl; Dabiré, Roch K.; Hopkins, Richard; Terenius, Olle

    2016-01-01

    The intolerable burden of malaria has for too long plagued humanity and the prospect of eradicating malaria is an optimistic, but reachable, target in the 21st century. However, extensive knowledge is needed about the spatial structure of mosquito populations in order to develop effective interventions against malaria transmission. We hypothesized that the microbiota associated with a mosquito reflects acquisition of bacteria in different environments. By analyzing the whole-body bacterial flora of An. gambiae mosquitoes from Burkina Faso by 16 S amplicon sequencing, we found that the different environments gave each mosquito a specific bacterial profile. In addition, the bacterial profiles provided precise and predicting information on the spatial dynamics of the mosquito population as a whole and showed that the mosquitoes formed clear local populations within a meta-population network. We believe that using microbiotas as proxies for population structures will greatly aid improving the performance of vector interventions around the world. PMID:26960555

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

    PubMed Central

    2013-01-01

    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. PMID:23336669

  19. Habitat discrimination by gravid Anopheles gambiae sensu lato – a push-pull system

    PubMed Central

    2014-01-01

    Background The non-random distribution of anopheline larvae in natural habitats suggests that gravid females discriminate between habitats of different quality. Whilst physical and chemical cues used by Culex and Aedes vector mosquitoes for selecting an oviposition site have been extensively studied, those for Anopheles remain poorly explored. Here the habitat selection by Anopheles gambiae sensu lato (s.l.), the principal African malaria vector, was investigated when presented with a choice of two infusions made from rabbit food pellets, or soil. Methods Natural colonization and larval survival was evaluated in artificial ponds filled randomly with either infusion. Dual-choice, egg-count bioassays evaluated the responses of caged gravid females to (1) two- to six-day old infusions versus lake water; (2) autoclaved versus non-autoclaved soil infusions; and assessed (3) the olfactory memory of gravid females conditioned in pellet infusion as larvae. Results Wild Anopheles exclusively colonized ponds with soil infusion and avoided those with pellet infusion. When the individual infusions were tested in comparison with lake water, caged An. gambiae sensu stricto (s.s.) showed a dose response: females increasingly avoided the pellet infusion with increasing infusion age (six-day versus lake water: odds ratio (OR) 0.22; 95% confidence interval (CI) 0.1-0.5) and showed increasing preference to lay eggs as soil infusion age increased (six-day versus lake water: OR 2.1; 95% CI 1.4-3.3). Larvae survived in soil infusions equally well as in lake water but died in pellet infusions. Anopheles gambiae s.s. preferred to lay eggs in the non-autoclaved soil (OR 2.6; 95% CI 1.8-3.7) compared with autoclaved soil. There was no change in the avoidance of pellet infusion by individuals reared in the infusion compared with those reared in lake water. Conclusion Wild and caged An. gambiae s.l. females discriminate between potential aquatic habitats for oviposition. These choices benefit

  20. Discriminative feeding behaviour of Anopheles gambiae s.s. on endemic plants in western Kenya

    PubMed Central

    Manda, H.; Gouagna, L. C.; Nyandat, E.; Kabiru, E. W.; Jackson, R. R.; Foster, W. A.; Githure, J. I.; Beier, J. C.; Hassanali, A.

    2009-01-01

    Anopheles gambiae Giles s.s. (Diptera: Culicidae) is known to feed on plant sugars, but this is the first experimental study to consider whether it discriminates between plant species. Thirteen perennial plant species were selected on the basis of their local availability within the vicinity of human dwellings and larval habitats of An. gambiae s.s. in western Kenya. Groups of 100 or 200 mosquitoes were released into cages either with a cutting of one plant type at a time (single-plant assay) or with cuttings of all 13 plants simultaneously (choice assay), respectively, and left overnight. In the choice assay, direct observations of the percentages of mosquitoes perching or feeding on each plant were recorded over four 1-h periods each night. For both types of assay, mosquitoes were recaptured and the percentage that had fed on plants was assessed by testing them individually for the presence of fructose. To identify which plants the choice-assay mosquitoes had fed on, gas chromatography (GC) profiles of samples of mosquito homogenates were compared with GC profiles of extracts from relevant parts of each plant. Four of the plants that were observed to have been fed on most frequently in the choice assay (Parthenium hysterophorus L., Tecoma stans L., Ricinus communis L., and Senna didymobotrya Fresen) were also shown to have been ingested most often by mosquitoes in both types of assay, suggesting that An. gambiae is differentially responsive to this range of plants, regardless of whether the plants were presented singly or mixed together. Significantly more females than males fed on plants, with the exception of P. hysterophorus L., one of the plants most frequently fed on. For most plant species (ten of 13), GC profiles indicated that An. gambiae obtained sugars primarily from flowers. The exceptions were P. hysterophorus L., Lantana camara L. and R. communis L., on which An. gambiae fed more often from leaves and stems than from flowers. PMID:17373953

  1. Crepuscular Behavioral Variation and Profiling of Opsin Genes in Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae)

    PubMed Central

    Jenkins, Adam M.; Muskavitch, Marc A. T.

    2015-01-01

    We understand little about photopreference and the molecular mechanisms governing vision-dependent behavior in vector mosquitoes. Investigations of the influence of photopreference on adult mosquito behaviors such as endophagy and exophagy and endophily and exophily will enhance our ability to develop and deploy vector-targeted interventions and monitoring techniques. Our laboratory-based analyses have revealed that crepuscular period photopreference differs between An. gambiae and An. stephensi. We employed qRT-PCR to assess crepuscular transcriptional expression patterns of long wavelength-, short wavelength-, and ultraviolet wavelength-sensing opsins (i.e., rhodopsin-class G-protein coupled receptors) in An. gambiae and in An. stephensi. Transcript levels do not exhibit consistent differences between species across diurnal cycles, indicating that differences in transcript abundances within this gene set are not correlated with these behavioral differences. Using developmentally staged and gender-specific RNAseq data sets in An. gambiae, we show that long wavelength-sensing opsins are expressed in two different patterns (one set expressed during larval stages, and one set expressed during adult stages), while short wavelength- and ultraviolet wavelength-sensing opsins exhibit increased expression during adult stages. Genomic organization of An. gambiae opsins suggests paralogous gene expansion of long wavelength-sensing opsins in comparison with An. stephensi. We speculate that this difference in gene number may contribute to variation between these species in photopreference behavior (e.g., visual sensitivity). PMID:26334802

  2. Crepuscular Behavioral Variation and Profiling of Opsin Genes in Anopheles gambiae and Anopheles stephensi (Diptera: Culicidae).

    PubMed

    Jenkins, Adam M; Muskavitch, Marc A T

    2015-05-01

    We understand little about photo-preference and the molecular mechanisms governing vision-dependent behavior in vector mosquitoes. Investigations of the influence of photo-preference on adult mosquito behaviors such as endophagy and exophagy and endophily and exophily will enhance our ability to develop and deploy vector-targeted interventions and monitoring techniques. Our laboratory-based analyses have revealed that crepuscular period photo-preference differs between An. gambiae and An. stephensi. We employed qRT-PCR to assess crepuscular transcriptional expression patterns of long wavelength-, short wavelength-, and ultraviolet wavelength-sensing opsins (i.e., rhodopsin-class G-protein coupled receptors) in An. gambiae and in An. stephensi. Transcript levels do not exhibit consistent differences between species across diurnal cycles, indicating that differences in transcript abundances within this gene set are not correlated with these behavioral differences. Using developmentally staged and gender-specific RNAseq data sets in An. gambiae, we show that long wavelength-sensing opsins are expressed in two different patterns (one set expressed during larval stages, and one set expressed during adult stages), while short wavelength- and ultraviolet wavelength-sensing opsins exhibit increased expression during adult stages. Genomic organization of An. gambiae opsins suggests paralogous gene expansion of long wavelength-sensing opsins in comparison with An. stephensi. We speculate that this difference in gene number may contribute to variation between these species in photo-preference behavior (e.g., visual sensitivity).

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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...

  4. Electroantennogram and behavioural responses of the malaria vector Anopheles gambiae to human-specific sweat components.

    PubMed

    Costantini, C; Birkett, M A; Gibson, G; Ziesmann, J; Sagnon, N F; Mohammed, H A; Coluzzi, M; Pickett, J A

    2001-09-01

    Afrotropical malaria vectors of the Anopheles gambiae complex (Diptera: Culicidae), particularly An. gambiae sensu stricto, are attracted mainly to human hosts. A major source of human volatile emissions is sweat, from which key human-specific components are the carboxylic acids (E)- and (Z)-3-methyl-2-hexenoic acid and 7-octenoic acid. Electrophysiological studies on the antennae of An. gambiae s.s. showed selective sensitivity to these compounds, with a threshold at 10(-6) g comparable to that of known olfactory stimulants 1-octen-3-ol, p-cresol, isovaleric acid, and lower than threshold sensitivity to L-lactic acid and the synthetic mosquito repellent N,N-diethyltoluamide (DEET). A combination of the acids released at concentrations > 10(-5) g in wind tunnel bioassays significantly reduced the response to CO2, the major attractant released by human hosts, for strains of An. gambiae s.s. originating from East and West Africa. Field trials with odour-baited entry traps (OBETs) in Burkina Faso showed that 7-octenoic acid significantly increased (by 1.7-fold) the catch of females of An. gambiae sensu lato (comprising two sibling species: An. arabiensis Patton and An. gambiae s.s.) in OBETs baited with CO2, whereas combinations of the acids significantly reduced the catch in CO2-baited traps (by 2.1-fold) and in whole human odour-baited traps (by 1.5-fold). The pure (E) and (Z) geometric isomers of 3-methyl-2-hexenoic acid gave comparable results to the (EIZ) isomer mixture. These results provide the first experimental evidence that human-specific compounds affect the behaviour of highly anthropophilic An. gambiae s.l. mosquitoes. The compounds appear to inhibit the upwind flight' response to known long-range attractants, and may serve either to mask' the attractants present or, more probably, to 'arrest' upwind flight when mosquitoes arrive at a host under natural conditions. In the final approach to hosts, vectors are known to reduce their flight speed and increase

  5. Assessment of alternative mating strategies in Anopheles gambiae: Does mating occur indoors?

    PubMed Central

    Dao, Adama; Adamou, Abdoulaye; Yaro, Alpha Seydou; Maïga, Hamidou Moussa; Kassogue, Yaya; Traoré, Sékou Fantamady; Lehmann, Tovi

    2016-01-01

    Mating in Anopheles gambiae has been observed only in outdoor swarms. Here we evaluate if mating also occurs indoors. Mark release recapture of virgin males and females in natural houses showed that mating occurred over a single day even when mosquitoes can leave the house through exit traps and without adaptation to laboratory conditions. In these experiments, insemination rate in the M molecular form of An. gambiae (and An. arabiensis) was higher than that of the S form (15% vs. 6%). Under these conditions, smaller females of the M form mated more frequently than larger females of that form. Sampling mosquitoes throughout the day showed that both sexes enter houses around sunrise and leave around sunset, staying indoors together from dawn to dusk. In an area dominated by the M form, the daily rate of insemination in samples from exit traps was approximately 5% higher than in those from entry traps, implying that mating occurred indoors. Importantly, frequency of cross mating between the molecular forms was as high as that between members of the same form, indicating that indoors - assortative mating breaks down. Altogether, these results suggest that indoor mating is an alternative mating strategy of the M molecular form of An. gambiae. Because naturally occurring mating couples have not yet been observed indoors, this conclusion awaits validation. PMID:18714863

  6. Visual and olfactory associative learning in the malaria vector Anopheles gambiae sensu stricto

    PubMed Central

    2012-01-01

    Background Memory and learning are critical aspects of the ecology of insect vectors of human pathogens because of their potential effects on contacts between vectors and their hosts. Despite this epidemiological importance, there have been only a limited number of studies investigating associative learning in insect vector species and none on Anopheline mosquitoes. Methods A simple behavioural assays was developed to study visual and olfactory associative learning in Anopheles gambiae, the main vector of malaria in Africa. Two contrasted membrane qualities or levels of blood palatability were used as reinforcing stimuli for bi-directional conditioning during blood feeding. Results Under such experimental conditions An. gambiae females learned very rapidly to associate visual (chequered and white patterns) and olfactory cues (presence and absence of cheese or Citronella smell) with the reinforcing stimuli (bloodmeal quality) and remembered the association for up to three days. Associative learning significantly increased with the strength of the conditioning stimuli used. Importantly, learning sometimes occurred faster when a positive reinforcing stimulus (palatable blood) was associated with an innately preferred cue (such as a darker visual pattern). However, the use of too attractive a cue (e.g. Shropshire cheese smell) was counter-productive and decreased learning success. Conclusions The results address an important knowledge gap in mosquito ecology and emphasize the role of associative memory for An. gambiae's host finding and blood-feeding behaviour with important potential implications for vector control. PMID:22284012

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

    PubMed Central

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

    1996-01-01

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

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

    PubMed

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

    2015-05-28

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

  9. Homing endonuclease mediated gene targeting in Anopheles gambiae cells and embryos.

    PubMed

    Windbichler, Nikolai; Papathanos, Philippos Aris; Catteruccia, Flaminia; Ranson, Hilary; Burt, Austin; Crisanti, Andrea

    2007-01-01

    Homing endonuclease genes (HEGs) are 'selfish' genetic elements that combine the capability to selectively disrupt specific gene sequences with the ability to rapidly spread from a few individuals to an entire population through homologous recombination repair events. Because of these properties, HEGs are regarded as promising candidates to transfer genetic modifications from engineered laboratory mosquitoes to wild-type populations including Anopheles gambiae the vector of human malaria. Here we show that I-SceI and I-PpoI homing endonucleases cleave their recognition sites with high efficiency in A. gambiae cells and embryos and we demonstrate HEG-induced homologous and non-homologous repair events in a variety of functional assays. We also propose a gene drive system for mosquitoes that is based on our finding that I-PpoI cuts genomic rDNA located on the X chromosome in A. gambiae, which could be used to selectively incapacitate X-carrying spermatozoa thereby imposing a severe male-biased sex ratio.

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

    PubMed

    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.

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

    PubMed Central

    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

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

    PubMed

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

    2015-06-01

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

  13. Daily rhythms in antennal protein and olfactory sensitivity in the malaria mosquito Anopheles gambiae

    PubMed Central

    Rund, Samuel S. C.; Bonar, Nicolle A.; Champion, Matthew M.; Ghazi, John P.; Houk, Cameron M.; Leming, Matthew T.; Syed, Zainulabeuddin; Duffield, Giles E.

    2013-01-01

    We recently characterized 24-hr daily rhythmic patterns of gene expression in Anopheles gambiae mosquitoes. These include numerous odorant binding proteins (OBPs), soluble odorant carrying proteins enriched in olfactory organs. Here we demonstrate that multiple rhythmically expressed genes including OBPs and takeout proteins, involved in regulating blood feeding behavior, have corresponding rhythmic protein levels as measured by quantitative proteomics. This includes AgamOBP1, previously shown as important to An. gambiae odorant sensing. Further, electrophysiological investigations demonstrate time-of-day specific differences in olfactory sensitivity of antennae to major host-derived odorants. The pre-dusk/dusk peaks in OBPs and takeout gene expression correspond with peak protein abundance at night, and in turn coincide with the time of increased olfactory sensitivity to odorants requiring OBPs and times of increased blood-feeding behavior. This suggests an important role for OBPs in modulating temporal changes in odorant sensitivity, enabling the olfactory system to coordinate with the circadian niche of An. gambiae. PMID:23986098

  14. Genome-Wide Patterns of Gene Expression during Aging in the African Malaria Vector Anopheles gambiae

    PubMed Central

    Wang, Mei-Hui; Marinotti, Osvaldo; James, Anthony A.; Walker, Edward; Githure, John; Yan, Guiyun

    2010-01-01

    The primary means of reducing malaria transmission is through reduction in longevity in days of the adult female stage of the Anopheles vector. However, assessing chronological age is limited to crude physiologic methods which categorize the females binomially as either very young (nulliparous) or not very young (parous). Yet the epidemiologically relevant reduction in life span falls within the latter category. Age-grading methods that delineate chronological age, using accurate molecular surrogates based upon gene expression profiles, will allow quantification of the longevity-reducing effects of vector control tools aimed at the adult, female mosquito. In this study, microarray analyses of gene expression profiles in the African malaria vector Anopheles gambiae were conducted during natural senescence of females in laboratory conditions. Results showed that detoxification-related and stress-responsive genes were up-regulated as mosquitoes aged. A total of 276 transcripts had age-dependent expression, independently of blood feeding and egg laying events. Expression of 112 (40.6%) of these transcripts increased or decreased monotonically with increasing chronologic age. Seven candidate genes for practical age assessment were tested by quantitative gene amplification in the An. gambiae G3 strain in a laboratory experiment and the Mbita strain in field enclosures set up in western Kenya under conditions closely resembling natural ones. Results were similar between experiments, indicating that senescence is marked by changes in gene expression and that chronological age can be gauged accurately and repeatedly with this method. These results indicate that the method may be suitable for accurate gauging of the age in days of field-caught, female An. gambiae. PMID:20967211

  15. Pyrethroid Resistance in Anopheles gambiae, in Bomi County, Liberia, Compromises Malaria Vector Control

    PubMed Central

    Temu, Emmanuel A.; Maxwell, Caroline; Munyekenye, Godwil; Howard, Annabel F. V.; Munga, Stephen; Avicor, Silas W.; Poupardin, Rodolphe; Jones, Joel J.; Allan, Richard; Kleinschmidt, Immo; Ranson, Hilary

    2012-01-01

    Background Long Lasting Insecticidal Nets (LLIN) and Indoor Residual Spraying (IRS) have both proven to be effective malaria vector control strategies in Africa and the new technology of insecticide treated durable wall lining (DL) is being evaluated. Sustaining these interventions at high coverage levels is logistically challenging and, furthermore, the increase in insecticide resistance in African malaria vectors may reduce the efficacy of these chemical based interventions. Monitoring of vector populations and evaluation of the efficacy of insecticide based control approaches should be integral components of malaria control programmes. This study reports on entomological survey conducted in 2011 in Bomi County, Liberia. Methods Anopheles gambiae larvae were collected from four sites in Bomi, Liberia, and reared in a field insectary. Two to five days old female adult An gambiae s.l. were tested using WHO tube (n = 2027) and cone (n = 580) bioassays in houses treated with DL or IRS. A sample of mosquitoes (n = 169) were identified to species/molecular form and screened for the presence of knock down resistance (kdr) alleles associated with pyrethroid resistance. Results Anopheles gambiae s.l tested were resistant to deltamethrin but fully susceptible to bendiocarb and fenithrothion. The corrected mortality of local mosquitoes exposed to houses treated with deltamethrin either via IRS or DL was 12% and 59% respectively, suggesting that resistance may affect the efficacy of these interventions. The presence of pyrethroid resistance was associated with a high frequency of the 1014F kdr allele (90.5%) although this mutation alone cannot explain the resistance levels observed. Conclusion High prevalence of resistance to deltamethrin in Bomi County may reduce the efficacy of malaria strategies relying on this class of insecticide. The findings highlight the urgent need to expand and sustain monitoring of insecticide resistance in Liberian malaria vectors

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

    NASA Astrophysics Data System (ADS)

    Rian, Sigrid Katrine Eivindsdatter

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

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

    PubMed

    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.

  18. Characterization of the target of ivermectin, the glutamate-gated chloride channel, from Anopheles gambiae

    PubMed Central

    Meyers, Jacob I.; Gray, Meg; Kuklinski, Wojtek; Johnson, Lucas B.; Snow, Christopher D.; Black, William C.; Partin, Kathryn M.; Foy, Brian D.

    2015-01-01

    ABSTRACT The use of insecticide-treated nets and indoor residual insecticides targeting adult mosquito vectors is a key element in malaria control programs. However, mosquito resistance to the insecticides used in these applications threatens malaria control efforts. Recently, the mass drug administration of ivermectin (IVM) has been shown to kill Anopheles gambiae mosquitoes and disrupt Plasmodium falciparum transmission in the field. We cloned the molecular target of IVM from A. gambiae, the glutamate-gated chloride channel (AgGluCl), and characterized its transcriptional patterns, protein expression and functional responses to glutamate and IVM. AgGluCl cloning revealed an unpredicted fourth splice isoform as well as a novel exon and splice site. The predicted gene products contained heterogeneity in the N-terminal extracellular domain and the intracellular loop region. Responses to glutamate and IVM were measured using two-electrode voltage clamp on Xenopus laevis oocytes expressing AgGluCl. IVM induced non-persistent currents in AgGluCl-a1 and did not potentiate glutamate responses. In contrast, AgGluCl-b was insensitive to IVM, suggesting that the AgGluCl gene could produce IVM-sensitive and -insensitive homomultimers from alternative splicing. AgGluCl isoform-specific transcripts were measured across tissues, ages, blood feeding status and sex, and were found to be differentially transcribed across these physiological variables. Lastly, we stained adult, female A. gambiae for GluCl expression. The channel was expressed in the antenna, Johnston's organ, supraesophageal ganglion and thoracic ganglia. In summary, we have characterized the first GluCl from a mosquito, A. gambiae, and described its unique activity and expression with respect to it as the target of the insecticide IVM. PMID:25994631

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

    PubMed Central

    OHBA, SHIN-YA; KAWADA, HITOSHI; DIDA, GABRIEL O.; JUMA, DUNCAN; SONYE, GORGE; MINAKAWA, NOBORU; TAKAGI, MASAHIRO

    2010-01-01

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

  20. Biochemical properties, expression profiles, and tissue localization of orthologous acetylcholinesterase-2 in the mosquito, Anopheles gambiae.

    PubMed

    Zhao, Picheng; Wang, Yang; Jiang, Haobo

    2013-03-01

    Acetylcholinesterases (AChEs) catalyze the hydrolysis of acetylcholine, a neurotransmitter for cholinergic neurotransmission in animals. Most insects studied so far possess two AChE genes: ace-1 paralogous and ace-2 orthologous to Drosophila melanogaster ace. We characterized the catalytic domain of Anopheles gambiae AChE1 in a previous study (Jiang et al., 2009) and report here biochemical properties of A. gambiae AChE2 expressed in Sf9 cells. An unknown protease in the expression system cleaved the recombinant AChE2 next to Arg(110), yielding two non-covalently associated polypeptides. A mixture of the intact and cleaved AChE2 had a specific activity of 72.3 U/mg, much lower than that of A. gambiae AChE1 (523 U/mg). The order of V(max)/K(M) values for the model substrates was acetylthiocholine > propionylthiocholine ≈ acetyl-(β-methyl)thiocholine > butyrylthiocholine. The IC(50)'s for eserine, carbaryl, BW284C51, paraoxon and malaoxon were 1.32, 13.6, 26.8, 192 and 294 nM, respectively. A. gambiae AChE2 bound eserine and carbaryl stronger than paraoxon and malaoxon, whereas eserine and malaoxon modified the active site Ser(232) faster than carbaryl or paraoxon did. Consequently, the k(i)'s were 1.173, 0.245, 0.029 and 0.018 μM(-1)min(-1) for eserine, carbaryl, paraoxon and malaoxon, respectively. Quantitative polymerase chain reactions showed a similar pattern of ace-1 and ace-2 expression. Their mRNAs were abundant in early embryos, greatly decreased in late embryos, larvae, pupae, and pharate adult, and became abundant again in adults. Both transcripts were higher in head and abdomen than thorax of adults and higher in male than female mosquitoes. Transcript levels of ace-1 were 1.9- to 361.8-fold higher than those of ace-2, depending on developmental stages and body parts. Cross-reacting polyclonal antibodies detected AChEs in adult brains, thoracic ganglia, and genital/rectal area. Activity assays, immunoblotting, and tandem mass spectrometric

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

    PubMed Central

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

    2013-01-01

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

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

    PubMed Central

    2013-01-01

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

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

    PubMed Central

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

    2002-01-01

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

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

    PubMed Central

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

    2015-01-01

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

  5. Visual arrestins in olfactory pathways of Drosophila and the malaria vector mosquito Anopheles gambiae

    PubMed Central

    Merrill, C. E.; Riesgo-Escovar, J.; Pitts, R. J.; Kafatos, F. C.; Carlson, J. R.; Zwiebel, L. J.

    2002-01-01

    Arrestins are important components for desensitization of G protein-coupled receptor cascades that mediate neurotransmission as well as olfactory and visual sensory reception. We have isolated AgArr1, an arrestin-encoding cDNA from the malaria vector mosquito, Anopheles gambiae, where olfaction is critical for vectorial capacity. Analysis of AgArr1 expression revealed an overlap between chemosensory and photoreceptor neurons. Furthermore, an examination of previously identified arrestins from Drosophila melanogaster exposed similar bimodal expression, and Drosophila arrestin mutants demonstrate impaired electrophysiological responses to olfactory stimuli. Thus, we show that arrestins in Drosophila are required for normal olfactory physiology in addition to their previously described role in visual signaling. These findings suggest that individual arrestins function in both olfactory and visual pathways in Dipteran insects; these genes may prove useful in the design of control strategies that target olfactory-dependent behaviors of insect disease vectors. PMID:11792843

  6. A modular chitin-binding protease associated with hemocytes and hemolymph in the mosquito Anopheles gambiae

    PubMed Central

    Danielli, Alberto; Loukeris, Thanasis G.; Lagueux, Marie; Müller, Hans-Michael; Richman, Adam; Kafatos, Fotis C.

    2000-01-01

    Sp22D, a modular serine protease encompassing chitin binding, low density lipoprotein receptor, and scavenger receptor cysteine-rich domains, was identified by molecular cloning in the malaria vector, Anopheles gambiae. It is expressed in multiple body parts and during much of development, most intensely in hemocytes. The protein appears to be posttranslationally modified. Its integral, putatively glycosylated form is secreted in the hemolymph, whereas a smaller form potentially generated by proteolytic processing is associated with the tissues. Bacterial challenge or wounding result in low-level RNA induction, but the protein does not bind to bacteria, nor is its processing affected by infection. However, Sp22D binds to chitin with high affinity and undergoes transient changes in processing during pupal to adult metamorphosis; it may respond to exposure to naked chitin during tissue remodeling or damage. PMID:10860981

  7. A modular chitin-binding protease associated with hemocytes and hemolymph in the mosquito Anopheles gambiae.

    PubMed

    Danielli, A; Loukeris, T G; Lagueux, M; Müller, H M; Richman, A; Kafatos, F C

    2000-06-20

    Sp22D, a modular serine protease encompassing chitin binding, low density lipoprotein receptor, and scavenger receptor cysteine-rich domains, was identified by molecular cloning in the malaria vector, Anopheles gambiae. It is expressed in multiple body parts and during much of development, most intensely in hemocytes. The protein appears to be posttranslationally modified. Its integral, putatively glycosylated form is secreted in the hemolymph, whereas a smaller form potentially generated by proteolytic processing is associated with the tissues. Bacterial challenge or wounding result in low-level RNA induction, but the protein does not bind to bacteria, nor is its processing affected by infection. However, Sp22D binds to chitin with high affinity and undergoes transient changes in processing during pupal to adult metamorphosis; it may respond to exposure to naked chitin during tissue remodeling or damage. PMID:10860981

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

    PubMed Central

    2011-01-01

    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 < 0.0001) and predator species (P < 0.0001) had a significant impact on the predation rate in the 24 hour evaluations. In semi-field experiments, predator species (P < 0.0001) and habitat type (P < 0.0001) were significant factors in both the daily survival and the overall developmental time of larvae. Pupation rates took significantly longer in habitats with refugia. 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

  9. Dihydroisoxazole inhibitors of Anopheles gambiae seminal transglutaminase AgTG3

    PubMed Central

    2014-01-01

    Background Current vector-based malaria control strategies are threatened by the rise of biochemical and behavioural resistance in mosquitoes. Researching mosquito traits of immunity and fertility is required to find potential targets for new vector control strategies. The seminal transglutaminase AgTG3 coagulates male Anopheles gambiae seminal fluids, forming a ‘mating plug’ that is required for male reproductive success. Inhibitors of AgTG3 can be useful both as chemical probes of A. gambiae reproductive biology and may further the development of new chemosterilants for mosquito population control. Methods A targeted library of 3-bromo-4,5-dihydroxoisoxazole inhibitors were synthesized and screened for inhibition of AgTG3 in a fluorescent, plate-based assay. Positive hits were tested for in vitro activity using cross-linking and mass spectrometry, and in vivo efficacy in laboratory mating assays. Results A targeted chemical library was screened for inhibition of AgTG3 in a fluorescent plate-based assay using its native substrate, plugin. Several inhibitors were identified with IC50 < 10 μM. Preliminary structure-activity relationships within the library support the stereo-specificity and preference for aromatic substituents in the chemical scaffold. Both inhibition of plugin cross-linking and covalent modification of the active site cysteine of AgTG3 were verified. Administration of an AgTG3 inhibitor to A. gambiae males by intrathoracic injection led to a 15% reduction in mating plug transfer in laboratory mating assays. Conclusions A targeted screen has identified chemical inhibitors of A. gambiae transglutaminase 3 (AgTG3). The most potent inhibitors are known inhibitors of human transglutaminase 2, suggesting a common binding pose may exist within the active site of both enzymes. Future efforts to develop additional inhibitors will provide chemical tools to address important biological questions regarding the role of the A. gambiae mating plug. A

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

    PubMed

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

    2016-11-01

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

  11. Anopheles gambiae Immune Responses to Human and Rodent Plasmodium Parasite Species

    PubMed Central

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

    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. 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 of invading

  12. Visualizing Non Infectious and Infectious Anopheles gambiae Blood Feedings in Naive and Saliva-Immunized Mice

    PubMed Central

    Choumet, Valerie; Attout, Tarik; Chartier, Loïc; Khun, Huot; Sautereau, Jean; Robbe-Vincent, Annie; Brey, Paul; Huerre, Michel; Bain, Odile

    2012-01-01

    Background Anopheles gambiae is a major vector of malaria and lymphatic filariasis. The arthropod-host interactions occurring at the skin interface are complex and dynamic. We used a global approach to describe the interaction between the mosquito (infected or uninfected) and the skin of mammals during blood feeding. Methods Intravital video microscopy was used to characterize several features during blood feeding. The deposition and movement of Plasmodium berghei sporozoites in the dermis were also observed. We also used histological techniques to analyze the impact of infected and uninfected feedings on the skin cell response in naive mice. Results The mouthparts were highly mobile within the skin during the probing phase. Probing time increased with mosquito age, with possible effects on pathogen transmission. Repletion was achieved by capillary feeding. The presence of sporozoites in the salivary glands modified the behavior of the mosquitoes, with infected females tending to probe more than uninfected females (86% versus 44%). A white area around the tip of the proboscis was observed when the mosquitoes fed on blood from the vessels of mice immunized with saliva. Mosquito feedings elicited an acute inflammatory response in naive mice that peaked three hours after the bite. Polynuclear and mast cells were associated with saliva deposits. We describe the first visualization of saliva in the skin by immunohistochemistry (IHC) with antibodies directed against saliva. Both saliva deposits and sporozoites were detected in the skin for up to 18 h after the bite. Conclusion This study, in which we visualized the probing and engorgement phases of Anopheles gambiae blood meals, provides precise information about the behavior of the insect as a function of its infection status and the presence or absence of anti-saliva antibodies. It also provides insight into the possible consequences of the inflammatory reaction for blood feeding and pathogen transmission. PMID

  13. Cytochrome P450 associated with insecticide resistance catalyzes cuticular hydrocarbon production in Anopheles gambiae

    PubMed Central

    Balabanidou, Vasileia; Kampouraki, Anastasia; MacLean, Marina; Blomquist, Gary J.; Tittiger, Claus; Juárez, M. Patricia; Mijailovsky, Sergio J.; Chalepakis, George; Anthousi, Amalia; Lynd, Amy; Antoine, Sanou; Hemingway, Janet; Ranson, Hilary; Lycett, Gareth J.; Vontas, John

    2016-01-01

    The role of cuticle changes in insecticide resistance in the major malaria vector Anopheles gambiae was assessed. The rate of internalization of 14C deltamethrin was significantly slower in a resistant strain than in a susceptible strain. Topical application of an acetone insecticide formulation to circumvent lipid-based uptake barriers decreased the resistance ratio by ∼50%. Cuticle analysis by electron microscopy and characterization of lipid extracts indicated that resistant mosquitoes had a thicker epicuticular layer and a significant increase in cuticular hydrocarbon (CHC) content (∼29%). However, the CHC profile and relative distribution were similar in resistant and susceptible insects. The cellular localization and in vitro activity of two P450 enzymes, CYP4G16 and CYP4G17, whose genes are frequently overexpressed in resistant Anopheles mosquitoes, were analyzed. These enzymes are potential orthologs of the CYP4G1/2 enzymes that catalyze the final step of CHC biosynthesis in Drosophila and Musca domestica, respectively. Immunostaining indicated that both CYP4G16 and CYP4G17 are highly abundant in oenocytes, the insect cell type thought to secrete hydrocarbons. However, an intriguing difference was indicated; CYP4G17 occurs throughout the cell, as expected for a microsomal P450, but CYP4G16 localizes to the periphery of the cell and lies on the cytoplasmic side of the cell membrane, a unique position for a P450 enzyme. CYP4G16 and CYP4G17 were functionally expressed in insect cells. CYP4G16 produced hydrocarbons from a C18 aldehyde substrate and thus has bona fide decarbonylase activity similar to that of dmCYP4G1/2. The data support the hypothesis that the coevolution of multiple mechanisms, including cuticular barriers, has occurred in highly pyrethroid-resistant An. gambiae. PMID:27439866

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

    PubMed

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

    2006-06-01

    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. 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 of invading

  15. Cytochrome P450 associated with insecticide resistance catalyzes cuticular hydrocarbon production in Anopheles gambiae.

    PubMed

    Balabanidou, Vasileia; Kampouraki, Anastasia; MacLean, Marina; Blomquist, Gary J; Tittiger, Claus; Juárez, M Patricia; Mijailovsky, Sergio J; Chalepakis, George; Anthousi, Amalia; Lynd, Amy; Antoine, Sanou; Hemingway, Janet; Ranson, Hilary; Lycett, Gareth J; Vontas, John

    2016-08-16

    The role of cuticle changes in insecticide resistance in the major malaria vector Anopheles gambiae was assessed. The rate of internalization of (14)C deltamethrin was significantly slower in a resistant strain than in a susceptible strain. Topical application of an acetone insecticide formulation to circumvent lipid-based uptake barriers decreased the resistance ratio by ∼50%. Cuticle analysis by electron microscopy and characterization of lipid extracts indicated that resistant mosquitoes had a thicker epicuticular layer and a significant increase in cuticular hydrocarbon (CHC) content (∼29%). However, the CHC profile and relative distribution were similar in resistant and susceptible insects. The cellular localization and in vitro activity of two P450 enzymes, CYP4G16 and CYP4G17, whose genes are frequently overexpressed in resistant Anopheles mosquitoes, were analyzed. These enzymes are potential orthologs of the CYP4G1/2 enzymes that catalyze the final step of CHC biosynthesis in Drosophila and Musca domestica, respectively. Immunostaining indicated that both CYP4G16 and CYP4G17 are highly abundant in oenocytes, the insect cell type thought to secrete hydrocarbons. However, an intriguing difference was indicated; CYP4G17 occurs throughout the cell, as expected for a microsomal P450, but CYP4G16 localizes to the periphery of the cell and lies on the cytoplasmic side of the cell membrane, a unique position for a P450 enzyme. CYP4G16 and CYP4G17 were functionally expressed in insect cells. CYP4G16 produced hydrocarbons from a C18 aldehyde substrate and thus has bona fide decarbonylase activity similar to that of dmCYP4G1/2. The data support the hypothesis that the coevolution of multiple mechanisms, including cuticular barriers, has occurred in highly pyrethroid-resistant An gambiae. PMID:27439866

  16. Life cycle transcriptome of the malaria mosquito Anopheles gambiae and comparison with the fruitfly Drosophila melanogaster.

    PubMed

    Koutsos, Anastasios C; Blass, Claudia; Meister, Stephan; Schmidt, Sabine; MacCallum, Robert M; Soares, Marcelo B; Collins, Frank H; Benes, Vladimir; Zdobnov, Evgeny; Kafatos, Fotis C; Christophides, George K

    2007-07-01

    The African mosquito Anopheles gambiae is the major vector of human malaria. We report a genome-wide survey of mosquito gene expression profiles clustered temporally into developmental programs and spatially into adult tissue-specific patterns. Global expression analysis shows that genes that belong to related functional categories or that encode the same or functionally linked protein domains are associated with characteristic developmental programs or tissue patterns. Comparative analysis of our data together with data published from Drosophila melanogaster reveal an overall strong and positive correlation of developmental expression between orthologous genes. The degree of correlation varies, depending on association of orthologs with certain developmental programs or functional groups. Interestingly, the similarity of gene expression is not correlated with the coding sequence similarity of orthologs, indicating that expression profiles and coding sequences evolve independently. In addition to providing a comprehensive view of temporal and spatial gene expression during the A. gambiae life cycle, this large-scale comparative transcriptomic analysis has detected important evolutionary features of insect transcriptomes.

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

    PubMed

    Lombardo, Fabrizio; Ghani, Yasmeen; Kafatos, Fotis C; Christophides, George K

    2013-01-01

    Reverse genetics in the mosquito Anopheles gambiae by RNAi mediated gene silencing has led in recent years to an advanced understanding of the mosquito immune response against infections with bacteria and malaria parasites. We developed RNAi screens in An. gambiae hemocyte-like cells using a library of double-stranded RNAs targeting 109 genes expressed highly or specifically in mosquito hemocytes to identify novel regulators of the hemocyte immune response. Assays included phagocytosis of bacterial bioparticles, expression of the antimicrobial peptide CEC1, and basal and induced expression of the mosquito complement factor LRIM1. A cell viability screen was also carried out to assess dsRNA cytotoxicity and to identify genes involved in cell growth and survival. Our results identify 22 novel immune regulators, including proteins putatively involved in phagosome assembly and maturation (Ca²⁺ channel, v-ATPase and cyclin-dependent protein kinase), pattern recognition (fibrinogen-domain lectins and Nimrod), immune modulation (peptidase and serine protease homolog), immune signaling (Eiger and LPS-induced factor), cell adhesion and communication (Laminin B1 and Ninjurin) and immune homeostasis (Lipophorin receptor). The development of robust functional cell-based assays paves the way for genome-wide functional screens to study the mosquito immune response to infections with human pathogens. PMID:23382679

  18. Analysis of the metabolome of Anopheles gambiae mosquito after exposure to Mycobacterium ulcerans

    PubMed Central

    Hoxmeier, J. Charles; Thompson, Brice D.; Broeckling, Corey D.; Small, Pamela; Foy, Brian D.; Prenni, Jessica; Dobos, Karen M.

    2015-01-01

    Infection with Mycobacterium ulcerans causes Buruli Ulcer, a neglected tropical disease. Mosquito vectors are suspected to participate in the transmission and environmental maintenance of the bacterium. However, mechanisms and consequences of mosquito contamination by M. ulcerans are not well understood. We evaluated the metabolome of the Anopheles gambiae mosquito to profile the metabolic changes associated with bacterial colonization. Contamination of mosquitoes with live M. ulcerans bacilli results in disruptions to lipid metabolic pathways of the mosquito, specifically the utilization of glycerolipid molecules, an affect that was not observed in mosquitoes exposed to dead M. ulcerans. These results are consistent with aberrations of lipid metabolism described in other mycobacterial infections, implying global host-pathogen interactions shared across diverse saprophytic and pathogenic mycobacterial species. This study implicates features of the bacterium, such as the putative M. ulcerans encoded phospholipase enzyme, which promote virulence, survival, and active adaptation in concert with mosquito development, and provides significant groundwork for enhanced studies of the vector-pathogen interactions using metabolomics profiling. Lastly, metabolic and survival data suggest an interaction which is unlikely to contribute to transmission of M. ulcerans by A. gambiae and more likely to contribute to persistence of M. ulcerans in waters cohabitated by both organisms. PMID:25784490

  19. Gene flow among populations of the malaria vector, Anopheles gambiae, in Mali, West Africa.

    PubMed Central

    Taylor, C; Touré, Y T; Carnahan, J; Norris, D E; Dolo, G; Traoré, S F; Edillo, F E; Lanzaro, G C

    2001-01-01

    The population structure of the Anopheles gambiae complex is unusual, with several sibling species often occupying a single area and, in one of these species, An. gambiae sensu stricto, as many as three "chromosomal forms" occurring together. The chromosomal forms are thought to be intermediate between populations and species, distinguishable by patterns of chromosome gene arrangements. The extent of reproductive isolation among these forms has been debated. To better characterize this structure we measured effective population size, N(e), and migration rates, m, or their product by both direct and indirect means. Gene flow among villages within each chromosomal form was found to be large (N(e)m > 40), was intermediate between chromosomal forms (N(e)m approximately 3-30), and was low between species (N(e)m approximately 0.17-1.3). A recently developed means for distinguishing among certain of the forms using PCR indicated rates of gene flow consistent with those observed using the other genetic markers. PMID:11156993

  20. Morphological Differentiation May Mediate Mate-Choice between Incipient Species of Anopheles gambiae s.s.

    PubMed Central

    Sanford, Michelle R.; Demirci, Berna; Marsden, Clare D.; Lee, Yoosook; Cornel, Anthony J.; Lanzaro, Gregory C.

    2011-01-01

    The M and S molecular forms of Anopheles gambiae s.s. have been considered incipient species for more than ten years, yet the mechanism underlying assortative mating of these incipient species has remained elusive. The discovery of the importance of harmonic convergence of wing beat frequency in mosquito mating and its relation to wing size have laid the foundation for exploring phenotypic divergence in wing size of wild populations of the two forms. In this study, wings from field collected mosquitoes were measured for wing length and wing width from two parts of the sympatric distribution, which differ with respect to the strength of assortative mating. In Mali, where assortative mating is strong, as evidenced by low rates of hybridization, mean wing lengths and wing widths were significantly larger than those from Guinea-Bissau. In addition, mean wing widths in Mali were significantly different between molecular forms. In Guinea-Bissau, assortative mating appears comparatively reduced and wing lengths and widths did not differ significantly between molecular forms. The data presented in this study support the hypothesis that wing beat frequency may mediate assortative mating in the incipient species of A. gambiae and represent the first documentation of a morphological difference between the M and S molecular forms. PMID:22132169

  1. Impact of trehalose transporter knockdown on Anopheles gambiae stress adaptation and susceptibility to Plasmodium falciparum infection

    PubMed Central

    Liu, Kun; Dong, Yuemei; Huang, Yuzheng; Rasgon, Jason L.; Agre, Peter

    2013-01-01

    Anopheles gambiae is a major vector mosquito for Plasmodium falciparum, the deadly pathogen causing most human malaria in sub-Saharan Africa. Synthesized in the fat body, trehalose is the predominant sugar in mosquito hemolymph. It not only provides energy but also protects the mosquito against desiccation and heat stresses. Trehalose enters the mosquito hemolymph by the trehalose transporter AgTreT1. In adult female A. gambiae, AgTreT1 is predominantly expressed in the fat body. We found that AgTreT1 expression is induced by environmental stresses such as low humidity or elevated temperature. AgTreT1 RNA silencing reduces the hemolymph trehalose concentration by 40%, and the mosquitoes succumb sooner after exposure to desiccation or heat. After an infectious blood meal, AgTreT1 RNA silencing reduces the number of P. falciparum oocysts in the mosquito midgut by over 70% compared with mock-injected mosquitoes. These data reveal important roles for AgTreT1 in stress adaptation and malaria pathogen development in a major vector mosquito. Thus, AgTreT1 may be a potential target for malaria vector control. PMID:24101462

  2. Antennal-Expressed Ammonium Transporters in the Malaria Vector Mosquito Anopheles gambiae

    PubMed Central

    Pulous, Fadi E.; Zwiebel, Laurence J.

    2014-01-01

    The principal Afrotropical malaria vector mosquito, Anopheles gambiae remains a significant threat to human health. In this anthropophagic species, females detect and respond to a range of human-derived volatile kairomones such as ammonia, lactic acid, and other carboxylic acids in their quest for blood meals. While the molecular underpinnings of mosquito olfaction and host seeking are becoming better understood, many questions remain unanswered. In this study, we have identified and characterized two candidate ammonium transporter genes, AgAmt and AgRh50 that are expressed in the mosquito antenna and may contribute to physiological and behavioral responses to ammonia, which is an important host kairomone for vector mosquitoes. AgAmt transcripts are highly enhanced in female antennae while a splice variant of AgRh50 appears to be antennal-specific. Functional expression of AgAmt in Xenopus laevis oocytes facilitates inward currents in response to both ammonium and methylammonium, while AgRh50 is able to partially complement a yeast ammonium transporter mutant strain, validating their conserved roles as ammonium transporters. We present evidence to suggest that both AgAmt and AgRh50 are in vivo ammonium transporters that are important for ammonia sensitivity in An. gambiae antennae, either by clearing ammonia from the sensillar lymph or by facilitating sensory neuron responses to environmental exposure. Accordingly, AgAmt and AgRh50 represent new and potentially important targets for the development of novel vector control strategies. PMID:25360676

  3. Conserved boundary elements from the Hox complex of mosquito, Anopheles gambiae.

    PubMed

    Ahanger, Sajad H; Srinivasan, Arumugam; Vasanthi, Dasari; Shouche, Yogesh S; Mishra, Rakesh K

    2013-01-01

    The conservation of hox genes as well as their genomic organization across the phyla suggests that this system of anterior-posterior axis formation arose early during evolution and has come under strong selection pressure. Studies in the split Hox cluster of Drosophila have shown that proper expression of hox genes is dependent on chromatin domain boundaries that prevent inappropriate interactions among different types of cis-regulatory elements. To investigate whether boundary function and their role in regulation of hox genes is conserved in insects with intact Hox clusters, we used an algorithm to locate potential boundary elements in the Hox complex of mosquito, Anopheles gambiae. Several potential boundary elements were identified that could be tested for their functional conservation. Comparative analysis revealed that like Drosophila, the bithorax region in A. gambiae contains an extensive array of boundaries and enhancers organized into domains. We analysed a subset of candidate boundary elements and show that they function as enhancer blockers in Drosophila. The functional conservation of boundary elements from mosquito in fly suggests that regulation of hox genes involving chromatin domain boundaries is an evolutionary conserved mechanism and points to an important role of such elements in key developmentally regulated loci.

  4. Ecophysiology of Anopheles gambiae s.l.: Persistence in the Sahel

    PubMed Central

    Huestis, Diana L.; Lehmann, Tovi

    2014-01-01

    The dry-season biology of malaria vectors is poorly understood, especially in arid environments when no surface waters are available for several months, such as during the dry season in the Sahel. Here we reappraise results on the dry-season physiology of members of the Anopheles gambiae s.l. complex in the broad context of dormancy in insects and especially in mosquitoes. We examine evidence on seasonal changes in reproduction, metabolism, stress tolerance, nutrition, molecular regulation, and environmental conditions and determine if the current results are compatible with dry-season diapause (aestivation) as the primary strategy for persistence throughout the dry season in the Sahel. In the process, we point out critical gaps in our knowledge that future studies can fill. We find compelling evidence that members of the An. gambiae s.l. complex undergo a form of aestivation during the Sahelian dry season by shifting energetic resources away from reproduction and towards increased longevity. Considering the differences between winter at temperate latitudes, which entails immobility of the insect and hence reliance on physiological solutions, as opposed to the Sahelian dry season, which restricts reproduction exclusively, we propose that behavioral changes play an important role in complementing physiological changes in this strategy. PMID:24933461

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

    PubMed

    Magnusson, Kalle; Mendes, Antonio M; Windbichler, Nikolai; Papathanos, Philippos-Aris; Nolan, Tony; Dottorini, Tania; Rizzi, Ermanno; Christophides, George K; Crisanti, Andrea

    2011-01-01

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

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

    PubMed

    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.

  7. Importance of algal biomass to growth and development of Anopheles gambiae larvae.

    PubMed

    Kaufman, Michael G; Wanja, Elizabeth; Maknojia, Shahnaz; Bayoh, M Nabie; Vulule, John M; Walker, Edward D

    2006-07-01

    We conducted experiments to investigate the importance of algal food resources for larval growth and adult emergence of Anopheles gambiae Giles s.s. in simulated larval habitats in Kenya, and in greenhouse and laboratory microcosms in the United States. In the first experiment, we used shading to reduce algal biomass, and because algal production and larval development might be a function of underlying soil nutrients, we crossed sun-shade treatments with soils of two distinct types collected near larval habitats. Shading reduced pupation rates and total adult biomass of An. gambiae by approximately 50%. Soil type had no significant effect on mosquito production, but it did significantly affect concentrations of phosphorus and chlorophyll a in the surface microlayer. In a subsequent experiment conducted in the greenhouse to reduce temperature differences found between the shaded and sunlit treatments, <1% of larvae in the shaded treatments reached the pupal stage. There was a marked reduction of chlorophyll a levels as a function of shading and larval density. In a third experiment, larvae receiving material harvested from sunlit surface microlayers performed as well as those receiving liver powder, whereas those receiving surface microlayer from shaded habitats suffered >90% mortality and failed to pupate. In a fourth experiment, glucose was added to shaded microcosms to stimulate bacterial activity in the absence of algae. Bacterial growth rates were 2 to 3 times higher, and larval development was enhanced in glucose-amended treatments. However, pupation rates and adult weights in glucose-amended shaded microcosms were still poor compared with those in nonamended sunlit microcosms. Overall, these results demonstrate the importance of algal biomass in the surface microlayers of larval habitats to development and adult production of An. gambiae.

  8. Functional development of carbon dioxide detection in the maxillary palp of Anopheles gambiae.

    PubMed

    Omondi, Bonaventure Aman; Majeed, Shahid; Ignell, Rickard

    2015-08-01

    Olfactory information drives several behaviours critical for the survival and persistence of insect pests and vectors. Insect behaviour is variable, linked to their biological needs, and regulated by physiological dynamics. For mosquitoes, CO2 is an important cue that signifies the presence of a host, and which elicits activation and attraction. To investigate the genetic basis of olfactory modulation in mosquitoes, we assayed changes in CO2 detection from receptor gene expression through physiological function to behaviour, associated with the onset of host seeking in the malaria vector, Anopheles gambiae. The gene encoding a subunit of the CO2 receptor, AgGr22, was found to be significantly up-regulated in host-seeking females, consistent with a significant increase in sensitivity of CO2-responsive neurons (cpA) housed in capitate peg sensilla of the maxillary palp. In addition, the odorant receptor AgOr28, which is expressed in cpC neurons, was significantly up-regulated. In contrast, AgOr8, which is expressed in cpB neurons, was not affected by this change in physiological state, in agreement with results for the obligate co-receptor Orco. Moreover, the sensitivity of the cpB neuron to (R)-1-octen-3-ol, a well-known mammalian kairomone, did not change in response to the onset of host seeking. The concentration of CO2 flux influenced both the propensity of A. gambiae to take off into the wind and the speed with which this activation occurred. Female A. gambiae mosquitoes responded to CO2 whether mature for host seeking or not, but onset of host seeking enhanced sensitivity and speed of activation at relevant doses of CO2.

  9. Antennal transcriptome profiles of anopheline mosquitoes reveal human host olfactory specialization in Anopheles gambiae

    PubMed Central

    2013-01-01

    Background Two sibling members of the Anopheles gambiae species complex display notable differences in female blood meal preferences. An. gambiae s.s. has a well-documented preference for feeding upon human hosts, whereas An. quadriannulatus feeds on vertebrate/mammalian hosts, with only opportunistic feeding upon humans. Because mosquito host-seeking behaviors are largely driven by the sensory modality of olfaction, we hypothesized that hallmarks of these divergent host seeking phenotypes will be in evidence within the transcriptome profiles of the antennae, the mosquito’s principal chemosensory appendage. Results To test this hypothesis, we have sequenced antennal mRNA of non-bloodfed females from each species and observed a number of distinct quantitative and qualitative differences in their chemosensory gene repertoires. In both species, these gene families show higher rates of sequence polymorphisms than the overall rates in their respective transcriptomes, with potentially important divergences between the two species. Moreover, quantitative differences in odorant receptor transcript abundances have been used to model potential distinctions in volatile odor receptivity between the two sibling species of anophelines. Conclusion This analysis suggests that the anthropophagic behavior of An. gambiae s.s. reflects the differential distribution of olfactory receptors in the antenna, likely resulting from a co-option and refinement of molecular components common to both species. This study improves our understanding of the molecular evolution of chemoreceptors in closely related anophelines and suggests possible mechanisms that underlie the behavioral distinctions in host seeking that, in part, account for the differential vectorial capacity of these mosquitoes. PMID:24182346

  10. Population genetic structure of Anopheles gambiae mosquitoes on Lake Victoria islands, west Kenya

    PubMed Central

    Chen, Hong; Minakawa, Noboru; Beier, John; Yan, Guiyun

    2004-01-01

    Background Understanding the genetic structure of island Anopheles gambiae populations is important for the current tactics in mosquito control and for the proposed strategy using genetically-modified mosquitoes (GMM). Genetically-isolated mosquito populations on islands are a potential site for testing GMM. The objective of this study was to determine the genetic structure of A. gambiae populations on the islands in Lake Victoria, western Kenya. Methods The genetic diversity and the population genetic structures of 13 A. gambiae populations from five islands on Lake Victoria and six villages from the surrounding mainland area in the Suba District were examined using six microsatellite markers. The distance range of sampling sites varied between 2.5 and 35.1 km. Results A similar level of genetic diversity between island mosquito populations and adjacent mainland populations was found. The average number of alleles per locus was 7.3 for the island populations and 6.8 for the mainland populations. The average observed heterozygosity was 0.32 and 0.28 for the island and mainland populations, respectively. A low but statistically significant genetic structure was detected among the island populations (FST = 0.019) and between the island and mainland populations (FST = 0.003). A total of 12 private alleles were found, and nine of them were from the island populations. Conclusion A level of genetic differentiation between the island and mainland populations was found. Large extent of gene flow between the island and mainland mosquito populations may result from wind- or human-assisted dispersal. Should the islands on Lake Victoria be used as a trial site for the release program of GMM, mosquito dispersal between the islands and between the island and the mainland should be vigorously monitored. PMID:15581429

  11. Functional development of carbon dioxide detection in the maxillary palp of Anopheles gambiae.

    PubMed

    Omondi, Bonaventure Aman; Majeed, Shahid; Ignell, Rickard

    2015-08-01

    Olfactory information drives several behaviours critical for the survival and persistence of insect pests and vectors. Insect behaviour is variable, linked to their biological needs, and regulated by physiological dynamics. For mosquitoes, CO2 is an important cue that signifies the presence of a host, and which elicits activation and attraction. To investigate the genetic basis of olfactory modulation in mosquitoes, we assayed changes in CO2 detection from receptor gene expression through physiological function to behaviour, associated with the onset of host seeking in the malaria vector, Anopheles gambiae. The gene encoding a subunit of the CO2 receptor, AgGr22, was found to be significantly up-regulated in host-seeking females, consistent with a significant increase in sensitivity of CO2-responsive neurons (cpA) housed in capitate peg sensilla of the maxillary palp. In addition, the odorant receptor AgOr28, which is expressed in cpC neurons, was significantly up-regulated. In contrast, AgOr8, which is expressed in cpB neurons, was not affected by this change in physiological state, in agreement with results for the obligate co-receptor Orco. Moreover, the sensitivity of the cpB neuron to (R)-1-octen-3-ol, a well-known mammalian kairomone, did not change in response to the onset of host seeking. The concentration of CO2 flux influenced both the propensity of A. gambiae to take off into the wind and the speed with which this activation occurred. Female A. gambiae mosquitoes responded to CO2 whether mature for host seeking or not, but onset of host seeking enhanced sensitivity and speed of activation at relevant doses of CO2. PMID:26056246

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

    PubMed

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

    2007-08-01

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

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

    PubMed

    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.

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

    PubMed

    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.

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

    PubMed Central

    Manda, Hortance; Gouagna, Louis C; Foster, Woodbridge A; Jackson, Robert R; Beier, John C; Githure, John I; Hassanali, Ahmed

    2007-01-01

    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 associated mainly with

  16. Effects of Plant-Community Composition on the Vectorial Capacity and Fitness of the Malaria Mosquito Anopheles gambiae

    PubMed Central

    Stone, Christopher M.; Jackson, Bryan T.; Foster, Woodbridge A.

    2012-01-01

    Dynamics of Anopheles gambiae abundance and malaria transmission potential rely strongly on environmental conditions. Female and male An. gambiae use sugar and are affected by its absence, but how the presence or absence of nectariferous plants affects An. gambiae abundance and vectorial capacity has not been studied. We report on four replicates of a cohort study performed in mesocosms with sugar-poor and sugar-rich plants, in which we measured mosquito survival, biting rates, and fecundity. Survivorship was greater with access to sugar-rich plant species, and mortality patterns were age-dependent. Sugar-poor populations experienced Weibull mortality patterns, and of four populations in the sugar-rich environment, two female and three male subpopulations were better fitted by Gompertz functions. A tendency toward higher biting rates in sugar-poor mesocosms, particularly for young females, was found. Therefore, vectorial capacity was pulled in opposing directions by nectar availability, resulting in highly variable vectorial capacity values. PMID:22927493

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

    PubMed Central

    Simard, Frédéric; Ayala, Diego; Kamdem, Guy Colince; Pombi, Marco; Etouna, Joachim; Ose, Kenji; Fotsing, Jean-Marie; Fontenille, Didier; Besansky, Nora J; Costantini, Carlo

    2009-01-01

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

  18. Gut Content Identification of Larvae of the Anopheles gambiae Complex in Western Kenya Using a Barcoding Approach

    PubMed Central

    Garros, C.; Ngungi, N.; Githeko, A.E.; Tuno, N.; Yan, G.

    2013-01-01

    Although larvae feeding and food source are vital to the development, survival and population regulation of African malaria vectors, the prey organisms of Anopheles gambiae larvae in the natural environment have not been well studied. This study used a molecular barcoding approach to investigate the natural diets of Anopheles gambiae s.l. larvae in Western Kenya. Gut contents from third- and fourth-instar larvae from natural habitats were dissected and DNA was extracted. The 18S ribosomal DNA gene was amplified, the resulting clones were screened using a RFLP method and non-mosquito clones were sequenced. Homology search and phylogenetic analyses were then conducted using the sequences of non-mosquito clones to identify the putative microorganisms ingested. The phylogenetic analyses clustered ingested microorganisms in four clades, including two clades of green algae (Chlorophyta, Chlorophyceae Class, Chlamydomonadales and Chlorococcales families), one fungal clade, and one unknown eukaryote clade. In parallel, using the same approach, an analysis of the biodiversity present in the larval habitats was done. This present study demonstrated the feasibility of the barcoding approach to infer the natural diets of Anopheles gambiae larvae. Our analysis suggests that despite the wide range of microorganisms available in natural habitats, mosquito larvae fed on specific groups of algae. The novel tools developed from this study can be used to improve our understanding of the larval ecology of African malaria vectors and to facilitate the development of new mosquito control tools. PMID:21585828

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

    PubMed

    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%.

  20. Identification of morphological and chemical markers of dry- and wet-season conditions in female Anopheles gambiae mosquitoes

    PubMed Central

    2014-01-01

    Background Increased understanding of the dry-season survival mechanisms of Anopheles gambiae in semi-arid regions could benefit vector control efforts by identifying weak links in the transmission cycle of malaria. In this study, we examined the effect of photoperiod and relative humidity on morphologic and chemical traits known to control water loss in mosquitoes. Methods Anopheles gambiae body size (indexed by wing length), mesothoracic spiracle size, and cuticular hydrocarbon composition (both standardized by body size) were examined in mosquitoes raised from eggs exposed to short photoperiod and low relative humidity, simulating the dry season, or long photoperiod and high relative humidity, simulating the wet-season. Results Mosquitoes exposed to short photoperiod exhibited larger body size and larger mesothoracic spiracle length than mosquitoes exposed to long photoperiod. Mosquitoes exposed to short photoperiod and low relative humidity exhibited greater total cuticular hydrocarbon amount than mosquitoes exposed to long photoperiod and high relative humidity. In addition, total cuticular hydrocarbon amount increased with age and was higher in mated females. Mean n-alkane retention time (a measure of cuticular hydrocarbon chain length) was lower in mosquitoes exposed to short photoperiod and low relative humidity, and increased with age. Individual cuticular hydrocarbon peaks were examined, and several cuticular hydrocarbons were identified as potential biomarkers of dry- and wet-season conditions, age, and insemination status. Conclusions Results from this study indicate that morphological and chemical changes underlie aestivation of Anopheles gambiae and may serve as biomarkers of aestivation. PMID:24970701

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

    PubMed Central

    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

  2. Effect of bioactive fractions of Citrullus vulgaris Schrad. leaf extract against Anopheles stephensi and Aedes aegypti.

    PubMed

    Mullai, K; Jebanesan, A; Pushpanathan, T

    2008-04-01

    The benzene extract of Citrullus vulgaris was tested against Anopheles stephensi and Aedes aegypti for the larvicidal activity and ovicidal properties. The crude benzene extract was found to be more effective against A. stephensi than A. aegypti. The LC50 values were 18.56 and 42.76 ppm respectively. The LC50 values for silica gel fractions (bioactive fractions I, II, III and IV) were 11.32, 14.12, 14.53 and 16.02 ppm respectively. The mean per cent hatchability of the egg rafts were observed after 48 h post treatment. The crude extract of benzene exerted 100% mortality at 250 ppm against A. stephensi and at 300 ppm against A. aegypti. The silica gel fractions I and II afforded 100% mortality at 100 ppm and III and IV exerted the hatchability rate of 4.9 and 5.3% at the same concentration against A. stephensi.

  3. Differences in timing of mating swarms in sympatric populations of Anopheles coluzzii and Anopheles gambiae s.s. (formerly An. gambiae M and S molecular forms) in Burkina Faso, West Africa

    PubMed Central

    2013-01-01

    Background The M and S molecular forms of Anopheles gambiae s.s. Giles appear to have speciated in West Africa and the M form is now formally named An. coluzzii Coetzee & Wilkerson sp.n. and the S form retains the nominotypical name (abbreviated here to An. gambiae). Reproductive isolation is thought to be the main barrier to hybridisation; even though both species are found in the same mating swarms, hybrid fertilisations in copulae have not been found in the study area. The aim of the study, therefore, was to determine whether differences in circadian and/or environmental control over the timing of swarming in the two species contribute to reproductive isolation. Methods The timing of male swarming in these species was recorded four nights per month over four years at five swarming sites in each of two villages. The timing of the start and end of swarming, and the concurrent environmental parameters, temperature, humidity and light intensity, were recorded for n = 20 swarms/month/species. The timing of 'spontaneous’ activity at dusk of individual An. coluzzii and An. gambiae males was video-recorded in an actograph outdoors for 21 nights. Results Of the environmental parameters considered, swarming was most strongly correlated with sunset (r2 > 0.946). Anopheles gambiae started and stopped swarming earlier than An. coluzzii (3:35 ± 0:68 min:sec and 4:51 ± 1:21, respectively), and the mean duration of swarming was 23:37 ± 0:33 for An. gambiae and 21:39 ± 0:33 for An. coluzzii. Accordingly, in principle, whenever both species swarm over the same marker, a mean of 15.3 ± 3.1% of An. gambiae swarming would occur before An. coluzzii males arrived, and 19.5 ± 4.55% of An. coluzzii swarming would occurred after An. gambiae males had stopped swarming. These results are consistent with the finding that An. gambiae males became active in the actograph 09:35 ± 00:22 min:sec earlier than An. coluzzii males. Conclusions The

  4. Intraspecific isozyme polymorphism of Anopheles gambiae in relation to environment, behavior, and malaria transmission in southwestern Burkina Faso.

    PubMed

    Coosemans, M; Smits, A; Roelants, P

    1998-01-01

    A cellulose acetate electrophoresis system was used to study the isozyme polymorphism of the Anopheles gambiae complex in a rural village and a city in southwestern Burkina Faso. In both areas A. gambiae Giles was the dominant species (95%) whereas A. arabiensis Patton represented only 5%. Both species were separated readily by octanol dehydrogenase Odh and mannose phosphate isomerase (Mpi) even if they shared some alleles at these two loci. Polymorphism analysis (13 loci) at the intraspecific level of A. gambiae showed a significant difference between the specimens collected in the city from those collected in the village in their allelic and genotypic frequencies of isocitrate dehydrogenase-1 and malate dehydrogenase-1 and in their allelic frequencies for Mpi. No genetic difference was observed between the human biting A. gambiae collected inside or outside the houses in either the village or the city. The Plasmodium falciparum-infected A. gambiae differed from the noninfected ones in their allelic and genotypic frequencies at Mpi and acid phosphatase (Acp). A two-fold difference in infection rate was found for the genotype Mpi130/130 and Acp110/100 compared with other genotypes. However, infected anophelines were found in all genotypes that belonged to these two enzyme systems. Consequently, no refractory mechanism occurs in these natural populations.

  5. Comparative and functional genomics of the innate immune system in the malaria vector Anopheles gambiae.

    PubMed

    Christophides, George K; Vlachou, Dina; Kafatos, Fotis C

    2004-04-01

    In much of Africa, the mosquito Anopheles gambiae is the major vector of human malaria, a devastating infectious disease caused by Plasmodium parasites. Vector and parasite interact at multiple stages and locations, and the nature and effectiveness of this reciprocal interaction determines the success of transmission. Many of the interactions engage the mosquito's innate immunity, a primitive but very effective defense system. In some cases, the mosquito kills the parasite, thus blocking the transmission cycle. However, not all interactions are antagonistic; some represent immune evasion. The sequence of the A. gambiae genome revealed numerous potential components of the innate immune system, and it established that they evolve rapidly, as summarized in the present review. Their rapid evolution by gene family expansion diversification as well as the prevalence of haplotype alleles in the best-studied families may reflect selective adaptation of the immune system to the exigencies of multiple immune challenges in a variety of ecologic niches. As a follow-up to the comparative genomic analysis, the development of functional genomic methodologies has provided novel opportunities for understanding the immune system and the nature of its interactions with the parasite. In this context, identification of both Plasmodium antagonists and protectors in the mosquito represents a significant conceptual advance. In addition to providing fundamental understanding of primitive immune systems, studies of mosquito interactions with the parasite open unprecedented opportunities for novel interventions against malaria transmission. The generation of transgenic mosquitoes that resist malaria infection in the wild and the development of antimalarial 'smart sprays' capable of disrupting interactions that are protective of the parasite, or reinforcing others that are antagonistic, represent technical challenges but also immense opportunities for improvement of global health.

  6. Delayed egg hatching of Anopheles gambiae (Diptera: Culicidae) pending water agitation.

    PubMed

    Ebrahimi, Babak; Shakibi, Sanam; Foster, Woodbridge A

    2014-05-01

    Mosquito eggs laid on water surfaces typically hatch spontaneously soon after the embryos within them become fully formed first-instar larvae. However, we have found that Anopheles gambiae Giles, an important vector of malaria in Africa, exhibits delayed hatching until the water surface is agitated, a feature overlooked in most laboratory colonies. Agitation within 24 h postoviposition, before embryonation was complete, failed to stimulate delayed postembryonic hatching of isolated eggs on the following day (day 2), when < 1% had hatched spontaneously. However, 5 min of water agitation of these dormant pharate first-instar larvae on day 2 resulted in an almost immediate hatch of 63.3 versus 0% of nonagitated controls, plus another 3.9 versus 0.3%, respectively, during the following 24 h. With daily agitation, installment hatching occurred mainly during 2-6 d postoviposition. The mean cumulative hatch after 7 d of daily agitation was 83.1 versus 1.1% of nonagitated eggs. Experiments with eggs in groups demonstrated that egg density and activity of already-hatched larvae had no stimulatory effect. Eggs stored 1-4 wk at 25.5 or at 15.5 degrees C, and then agitated daily for 6 d at 25.5 degrees C, showed a gradual decline in viability. Viability was sustained longer at the lower temperature. Implications of agitation-induced egg hatching for rainy-season and dry-season ecology of An. gambiae are discussed. Suspended hatching and cool storage already are proving convenient for efficient mass rearing and accurate modeling of weather-based population dynamics.

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

    PubMed Central

    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.

    2011-01-01

    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 – 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 – 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. PMID:22198355

  8. Seasonal variation in metabolic rate, flight activity and body size of Anopheles gambiae in the Sahel

    PubMed Central

    Huestis, Diana L.; Yaro, Alpha S.; Traoré, Adama I.; Dieter, Kathryne L.; Nwagbara, Juliette I.; Bowie, Aleah C.; Adamou, Abdoulaye; Kassogué, Yaya; Diallo, Moussa; Timbiné, Seydou; Dao, Adama; Lehmann, Tovi

    2012-01-01

    SUMMARY Malaria in Africa is vectored primarily by the Anopheles gambiae complex. Although the mechanisms of population persistence during the dry season are not yet known, targeting dry season mosquitoes could provide opportunities for vector control. In the Sahel, it appears likely that M-form A. gambiae survive by aestivation (entering a dormant state). To assess the role of eco-physiological changes associated with dry season survival, we measured body size, flight activity and metabolic rate of wild-caught mosquitoes throughout 1 year in a Sahelian locality, far from permanent water sources, and at a riparian location adjacent to the Niger River. We found significant seasonal variation in body size at both the Sahelian and riparian sites, although the magnitude of the variation was greater in the Sahel. For flight activity, significant seasonality was only observed in the Sahel, with increased flight activity in the wet season when compared with that just prior to and throughout the dry season. Whole-organism metabolic rate was affected by numerous biotic and abiotic factors, and a significant seasonal component was found at both locations. However, assay temperature accounted completely for seasonality at the riparian location, while significant seasonal variation remained after accounting for all measured variables in the Sahel. Interestingly, we did not find that mean metabolic rate was lowest during the dry season at either location, contrary to our expectation that mosquitoes would conserve energy and increase longevity by reducing metabolism during this time. These results indicate that mosquitoes may use mechanisms besides reduced metabolic rate to enable survival during the Sahelian dry season. PMID:22623189

  9. Gene Flow–Dependent Genomic Divergence between Anopheles gambiae M and S Forms

    PubMed Central

    Weetman, David; Wilding, Craig S.; Steen, Keith; Pinto, João; Donnelly, Martin J.

    2012-01-01

    Anopheles gambiae sensu stricto exists as two often-sympatric races termed the M and S molecular forms, characterized by fixed differences at an X-linked marker. Extreme divergence between M and S forms at pericentromeric “genomic islands” suggested that selection on variants therein could be driving interform divergence in the presence of ongoing gene flow, but recent work has detected much more widespread genomic differentiation. Whether such genomic islands are important in reproductive isolation or represent ancestral differentiation preserved by low recombination is currently unclear. A critical test of these competing hypotheses could be provided by comparing genomic divergence when rates of recent introgression vary. We genotyped 871 single nucleotide polymorphisms (SNPs) in A. gambiae sensu stricto from locations of M and S sympatry and allopatry, encompassing the full range of observed hybridization rates (0–25%). M and S forms were readily partitioned based on genomewide SNP variation in spite of evidence for ongoing introgression that qualitatively reflects hybridization rates. Yet both the level and the heterogeneity of genomic divergence varied markedly in line with levels of introgression. A few genomic regions of differentiation between M and S were common to each sampling location, the most pronounced being two centromere–proximal speciation islands identified previously but with at least one additional region outside of areas expected to exhibit reduced recombination. Our results demonstrate that extreme divergence at genomic islands does not simply represent segregating ancestral polymorphism in regions of low recombination and can be resilient to substantial gene flow. This highlights the potential for islands comprising a relatively small fraction of the genome to play an important role in early-stage speciation when reproductive isolation is limited. PMID:21836185

  10. Expression of metallothionein and α-tubulin in heavy metal-tolerant Anopheles gambiae sensu stricto (Diptera: Culicidae)

    PubMed Central

    Mireji, Paul O.; Keating, Joseph; Hassanali, Ahmed; Impoinvil, Daniel E.; Mbogo, Charles M.; Njeri, Martha; Nyambaka, Hudson; Kenya, Eucharia; Githure, John I; Beier, John C.

    2009-01-01

    Anopheles mosquitoes have been shown to adapt to heavy metals in their natural habitats. In this study we explored the possibility of using Anopheles gambiae sensu stricto as bio-reporters for environmental heavy metal pollution through expressions of their metal responsive metallothionein and α-tubulin genes. The study was undertaken with third instar larvae after selection by cadmium, copper, or lead at LC30 through five successive generations. Expression levels were determined in the fifth generation by semi quantitative RT-PCR on the experimental and control populations. The data were analyzed using one-way ANOVA. The highest metallothionein (F3, 11= 4.574, P = 0.038) and α-tubulin (F3,11= 12.961, P = 0.002) responses were observed in cadmium-tolerant treatments. There was significantly higher expression of metallothionein in cadmium or copper treatments relative to the control (P = 0.012), and in cadmium than in lead treatments (P = 0.044). Expressions of α-tubulin were significantly higher in cadmium than in control treatments (P = 0.008). These results demonstrate capacity of An. gambiae s.s. to develop tolerance to increased levels of heavy metal challenge. The results also confirm the potential of heavy metal responsive genes in mosquitoes as possible bio-indicators of heavy metal environmental pollution. How the tolerance and expressions relate to An. gambiae s.s. fitness and vectorial capacity in the environment remains to be elucidated. PMID:19735939

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

    PubMed

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

    2014-04-22

    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

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

    PubMed Central

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

    2014-01-01

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

  13. Comparative Genome and Proteome Analysis of Anopheles gambiae and Drosophila melanogaster

    NASA Astrophysics Data System (ADS)

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

    2002-10-01

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

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

    PubMed

    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.

  15. The neurotransmitters serotonin and glutamate accelerate the heart rate of the mosquito Anopheles gambiae.

    PubMed

    Hillyer, Julián F; Estévez-Lao, Tania Y; Mirzai, Homa E

    2015-10-01

    Serotonin and glutamate are neurotransmitters that in insects are involved in diverse physiological processes. Both serotonin and glutamate have been shown to modulate the physiology of the dorsal vessel of some insects, yet until the present study, their activity in mosquitoes remained unknown. To test whether serotonin or glutamate regulate dorsal vessel physiology in the African malaria mosquito, Anopheles gambiae, live mosquitoes were restrained, and a video of the contracting heart (the abdominal portion of the dorsal vessel) was acquired. These adult female mosquitoes were then injected with various amounts of serotonin, glutamate, or a control vehicle solution, and additional videos were acquired at 2 and 10 min post-treatment. Comparison of the videos taken before and after treatment revealed that serotonin accelerates the frequency of heart contractions, with the cardioacceleration being significantly more pronounced when the wave-like contractions of cardiac muscle propagate in the anterograde direction (toward the head). Comparison of the videos taken before and after treatment with glutamate revealed that this molecule is also cardioacceleratory. However, unlike serotonin, the activity of glutamate does not depend on whether the contractions propagate in the anterograde or the retrograde (toward the posterior of the abdomen) directions. Serotonin or glutamate induces a minor change or no change in the percentage of contractions and the percentage of the time that the heart contracts in the anterograde or the retrograde directions. In summary, this study shows that the neurotransmitters serotonin and glutamate increase the heart contraction rate of mosquitoes. PMID:26099947

  16. The neurotransmitters serotonin and glutamate accelerate the heart rate of the mosquito Anopheles gambiae.

    PubMed

    Hillyer, Julián F; Estévez-Lao, Tania Y; Mirzai, Homa E

    2015-10-01

    Serotonin and glutamate are neurotransmitters that in insects are involved in diverse physiological processes. Both serotonin and glutamate have been shown to modulate the physiology of the dorsal vessel of some insects, yet until the present study, their activity in mosquitoes remained unknown. To test whether serotonin or glutamate regulate dorsal vessel physiology in the African malaria mosquito, Anopheles gambiae, live mosquitoes were restrained, and a video of the contracting heart (the abdominal portion of the dorsal vessel) was acquired. These adult female mosquitoes were then injected with various amounts of serotonin, glutamate, or a control vehicle solution, and additional videos were acquired at 2 and 10 min post-treatment. Comparison of the videos taken before and after treatment revealed that serotonin accelerates the frequency of heart contractions, with the cardioacceleration being significantly more pronounced when the wave-like contractions of cardiac muscle propagate in the anterograde direction (toward the head). Comparison of the videos taken before and after treatment with glutamate revealed that this molecule is also cardioacceleratory. However, unlike serotonin, the activity of glutamate does not depend on whether the contractions propagate in the anterograde or the retrograde (toward the posterior of the abdomen) directions. Serotonin or glutamate induces a minor change or no change in the percentage of contractions and the percentage of the time that the heart contracts in the anterograde or the retrograde directions. In summary, this study shows that the neurotransmitters serotonin and glutamate increase the heart contraction rate of mosquitoes.

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

    PubMed

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

    2016-03-01

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

  18. Crystal structure of native Anopheles gambiae Serpin-2, a negative regulator of melanization in mosquitoes

    PubMed Central

    An, Chunju; Lovell, Scott; Kanost, Michael R.; Battaile, Kevin P.; Michel, Kristin

    2011-01-01

    Serpins are the dominant group of protease inhibitors in metazoans that control a wide variety of biological processes including major innate immune reactions. One of these inhibitors, SRPN2, controls melanization in mosquitoes – a powerful, arthropod-specific innate immune response. SRPN2 depletion from the hemolymph of adult female mosquitoes significantly reduces longevity and therefore this serpin is a potential target for novel insecticides. We report here the crystal structure of SRPN2 in its native conformation from the African malaria mosquito, Anopheles gambiae to 1.75 Å resolution. SRPN2 adopts a similar fold as observed for other serpins with a core of three β-sheets surrounded by nine α-helices with an exposed reactive center loop (RCL) that extends from the protein body. Similar to other native serpin structures, several residues within the reactive center loop were disordered and could not be modeled. Intriguingly, the N-terminal hinge of the RCL in SRPN2 was found to be inserted into β-sheet A, suggesting a potential activation mechanism analogous to heparin-mediated activation of Antithrombin III. PMID:21465556

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

    PubMed

    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.

  20. Survivorship and distribution of immature Anopheles gambiae s.l. (Diptera: Culicidae) in Banambani village, Mali.

    PubMed

    Edillo, Frances E; Touré, Yeya T; Lanzaro, Gregory C; Dolo, Guimogo; Taylor, Charles E

    2004-05-01

    We observed the survivorship and distribution of larvae and pupae of Anopheles gambiae s.l. Giles immature stages in three habitats (rock pools, swamp, and puddles) in Banambani village. Mali, West Africa, during the mid-rainy season of 2000. Horizontal life tables were constructed for immatures in the laboratory. Times spent in the various immature stages were determined, and laboratory survival was measured. Vertical life tables were obtained from each habitat. We found large day-to-day variation for age class composition within habitats across days. The swamp samples had small but statistically significant different distributions in some instar stages compared with rock pools and puddles as affected by precipitation history. There were obviously unstable age distributions in the swamp and puddles and to some extent in rock pools. There were more individuals in some later age classes than in earlier ones. The daily survival estimates using an exponential decay model were 0.807 in rock pools, 0.899 in the swamp, 0.818 in puddles, and 0.863 in the overall village. Possible reasons for the departure from stable age distribution were cannibalism, predation and other complex interactions, rainfall effects, sampling bias, and differences in physicochemical properties of the water in the habitats. PMID:15185933

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

    PubMed

    Wang, Ying; Gilbreath, Thomas M; Kukutla, Phanidhar; Yan, Guiyun; Xu, Jiannong

    2011-01-01

    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.

  2. Carbamate and pyrethroid resistance in the akron strain of Anopheles gambiae.

    PubMed

    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

    2015-06-01

    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

  3. Fumigant toxicity of the essential oils of some African plants against Anopheles gambiae sensu stricto.

    PubMed

    Omolo, M O; Okinyo, D; Ndiege, I O; Lwande, W; Hassanali, A

    2005-03-01

    The essential oils from 15 species of African plants selected by ethnobotanical considerations and field inspection (odour and presence of insects) were screened for fumigant toxicity to Anopheles gambiae s.s. in the laboratory. Essential oils from 6 species showed varying levels of toxicity, with Conyza newii (Compositae) and Plectranthus marruboides (Labiateae) being the most potent. Fifty compounds representing approximately 74% of the essential oil of C. newii were identified by GC-MS and GC-coinjection (for available standards). The major and some of the minor constituents of the two oils were assayed at different doses. Two compounds, from C. newii, perillaldehyde and perillyl alcohol, exhibited higher fumigant toxicity (LD50 = 1.05 x 10(-4) and 2.52 x 10(-4) mg cm(-3), respectively) than the parent oil (2.0 x 10(-3) mg cm(-3)). GC-MS analysis of the essential oil of P. marruboides gave results similar to that previously reported. Interestingly, none of its components were active, suggesting that the insecticidal activity of the oil results from either some of the minor components or as a blend effect of some of the major constituents. PMID:15830848

  4. Interactive cost of Plasmodium infection and insecticide resistance in the malaria vector Anopheles gambiae

    PubMed Central

    Alout, Haoues; Dabiré, Roch K.; Djogbénou, Luc S.; Abate, Luc; Corbel, Vincent; Chandre, Fabrice; Cohuet, Anna

    2016-01-01

    Insecticide resistance raises concerns for the control of vector-borne diseases. However, its impact on parasite transmission could be diverse when considering the ecological interactions between vector and parasite. Thus we investigated the fitness cost associated with insecticide resistance and Plasmodium falciparum infection as well as their interactive cost on Anopheles gambiae survival and fecundity. In absence of infection, we observed a cost on fecundity associated with insecticide resistance. However, survival was higher for mosquito bearing the kdr mutation and equal for those with the ace-1R mutation compared to their insecticide susceptible counterparts. Interestingly, Plasmodium infection reduced survival only in the insecticide resistant strains but not in the susceptible one and infection was associated with an increase in fecundity independently of the strain considered. This study provides evidence for a survival cost associated with infection by Plasmodium parasite only in mosquito selected for insecticide resistance. This suggests that the selection of insecticide resistance mutation may have disturbed the interaction between parasites and vectors, resulting in increased cost of infection. Considering the fitness cost as well as other ecological aspects of this natural mosquito-parasite combination is important to predict the epidemiological impact of insecticide resistance. PMID:27432257

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

    PubMed Central

    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

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

    PubMed

    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.

  7. Identification and characterization of two chitin synthase genes in African malaria mosquito, Anopheles gambiae

    PubMed Central

    Zhang, Xin; Zhang, Jianzhen; Park, Yoonseong; Zhu, Kun Yan

    2012-01-01

    Chitin synthase (CHS) represents an attractive target site for combating insect pests as insect growth and development are strictly dependent on precisely tuned chitin biosynthesis and this pathway is absent in humans and other vertebrates. Current knowledge on CHS in insects, especially their structures, functions, and regulations is still very limited. We report the identification and characterization of two chitin synthase genes, AgCHS1 and AgCHS2, in African malaria mosquito, Anopheles gambiae. AgCHS1 and AgCHS2 were predicted to encode proteins of 1,578 and 1,586 amino acid residues, respectively. Their deduced amino acid sequences show high similarities to other insect chitin synthases. Transcriptional analysis indicated that AgCHS1 was expressed in egg, larval, pupal and adult stages whereas AgCHS2 appeared to be expressed at relatively low levels, particularly during the larval stages as examined by reverse transcription (RT)-PCR and real-time quantitative PCR. Relatively high expression was detected in the carcass followed by the foregut and hindgut for AgCHS1, and the foregut (cardia included) followed by the midgut for AgCHS2. Fluorescence in situ hybridization (FISH) and immunohistochemical analysis revealed new information including the localization of the two enzymes in the ommatidia of the compound eyes, and AgCHS2 in the thoracic and abdominal inter-segmental regions of pupal integument. PMID:22683441

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

    PubMed Central

    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

  9. Survivorship and distribution of immature Anopheles gambiae s.l. (Diptera: Culicidae) in Banambani village, Mali.

    PubMed

    Edillo, Frances E; Touré, Yeya T; Lanzaro, Gregory C; Dolo, Guimogo; Taylor, Charles E

    2004-05-01

    We observed the survivorship and distribution of larvae and pupae of Anopheles gambiae s.l. Giles immature stages in three habitats (rock pools, swamp, and puddles) in Banambani village. Mali, West Africa, during the mid-rainy season of 2000. Horizontal life tables were constructed for immatures in the laboratory. Times spent in the various immature stages were determined, and laboratory survival was measured. Vertical life tables were obtained from each habitat. We found large day-to-day variation for age class composition within habitats across days. The swamp samples had small but statistically significant different distributions in some instar stages compared with rock pools and puddles as affected by precipitation history. There were obviously unstable age distributions in the swamp and puddles and to some extent in rock pools. There were more individuals in some later age classes than in earlier ones. The daily survival estimates using an exponential decay model were 0.807 in rock pools, 0.899 in the swamp, 0.818 in puddles, and 0.863 in the overall village. Possible reasons for the departure from stable age distribution were cannibalism, predation and other complex interactions, rainfall effects, sampling bias, and differences in physicochemical properties of the water in the habitats.

  10. Behavioral responses of catnip (Nepeta cataria) by two species of mosquitoes, Aedes aegypti and Anopheles harrisoni, in Thailand.

    PubMed

    Polsomboon, Suppaluck; Grieco, John P; Achee, Nicole L; Chauhan, Kamlesh R; Tanasinchayakul, Somchai; Pothikasikorn, Jinrapa; Chareonviriyaphap, Theeraphap

    2008-12-01

    An investigation of the biological effect of catnip oil (Nepeta cataria L.) on the behavioral response of field collected Aedes aegypti and Anopheles harrisoni was conducted using an automated excitorepellency test system. Aedes aegypti showed significantly higher escape rates from the contact chamber at 5% catnip oil compared to other concentrations (P < 0.05). With Anopheles harrisoni, a high escape response was seen at 2.5% catnip oil from the contact chamber, while in the noncontact chamber a higher escape response was observed at a concentration of 5%. Results showed that this compound exhibits both irritant and repellent actions.

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

    PubMed

    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.

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

    PubMed Central

    2012-01-01

    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. PMID:23206364

  13. Select Small Core Structure Carbamates Exhibit High Contact Toxicity to “Carbamate-Resistant” Strain Malaria Mosquitoes, Anopheles gambiae (Akron)

    PubMed Central

    Wong, Dawn M.; Li, Jianyong; Chen, Qiao-Hong; Han, Qian; Mutunga, James M.; Wysinski, Ania; Anderson, Troy D.; Ding, Haizhen; Carpenetti, Tiffany L.; Verma, Astha; Islam, Rafique; Paulson, Sally L.; Lam, Polo C.-H.; Totrov, Maxim; Bloomquist, Jeffrey R.; Carlier, Paul R.

    2012-01-01

    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 (kcat) 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 (LC50>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 (LC50 = 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. PMID:23049714

  14. Toxicity of Boswellia dalzielii (Burseraceae) Leaf Fractions Against Immature Stages of Anopheles gambiae (Giles) and Culex quinquefasciatus (Say) (Diptera: Culicidae)

    PubMed Central

    Younoussa, Lame; Nukenine, Elias Nchiwan; Esimone, Charles Okechukwu

    2016-01-01

    Mosquitoes are vectors of several human pathogens, and great attention has recently been placed on insecticides from plant-derived products, in search for mosquito control agents. This study, thus, investigated the potency of Boswellia dalzielii methanol leaf extract and its four fractions as mosquito ovicide, larvicide, and pupicide against Anopheles gambiae and Culex quinquefasciatus. The plant products were tested at the following concentrations: 125, 250, 500, 1000, and 2000 ppm on eggs and 312.5, 625, 1250, and 2500 ppm on the larvae and pupae of the mosquitoes. For results, hatchability of A. gambiae eggs was reduced to 5% with n-hexane fraction at 2000 ppm. Among the plant products tested, n-hexane fraction was most toxic against A. gambiae (LC50 = 385.9 ppm) and C. quinquefasciatus (LC50 = 3394.9 ppm). The n-hexane fraction of B. dalzielii might be used as a mosquitocidal agent in the breeding sites of A. gambiae and C. quinquefasciatus. PMID:27279752

  15. Toxicity of Boswellia dalzielii (Burseraceae) Leaf Fractions Against Immature Stages of Anopheles gambiae (Giles) and Culex quinquefasciatus (Say) (Diptera: Culicidae).

    PubMed

    Younoussa, Lame; Nukenine, Elias Nchiwan; Esimone, Charles Okechukwu

    2016-01-01

    Mosquitoes are vectors of several human pathogens, and great attention has recently been placed on insecticides from plant-derived products, in search for mosquito control agents. This study, thus, investigated the potency of Boswellia dalzielii methanol leaf extract and its four fractions as mosquito ovicide, larvicide, and pupicide against Anopheles gambiae and Culex quinquefasciatus. The plant products were tested at the following concentrations: 125, 250, 500, 1000, and 2000 ppm on eggs and 312.5, 625, 1250, and 2500 ppm on the larvae and pupae of the mosquitoes. For results, hatchability of A. gambiae eggs was reduced to 5% with n-hexane fraction at 2000 ppm. Among the plant products tested, n-hexane fraction was most toxic against A. gambiae (LC50 = 385.9 ppm) and C. quinquefasciatus (LC50 = 3394.9 ppm). The n-hexane fraction of B. dalzielii might be used as a mosquitocidal agent in the breeding sites of A. gambiae and C. quinquefasciatus. PMID:27279752

  16. A theoretical model of the dynamics of an Anopheles gambiae population under challenge with eggs giving rise to sterile males*

    PubMed Central

    Cuellar, C. B.

    1969-01-01

    An explanation is given of the probable effects of seeding a breeding place of Anopheles gambiae with hybrid eggs that produce almost exclusively sterile males. For the calculations, the dynamics of an A. gambiae population in a single breeding place have been simulated in a computer programme. General considerations and implications are discussed with reference to practical applications, and the conclusion is reached that it will be feasible to eradicate the species from any breeding place only if the number of factory-produced eggs distributed daily bears a known, optimal relationship to the numbers deposited daily on the site. The period of treatment, if the first-day ratio of normal: factory-produced eggs is 1: 1, was estimated to be in the region of 9 weeks. Several runs of the computer have been condensed in a graph which might be used in the early evaluation of progress in the field. PMID:5306541

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

    PubMed

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

    2002-09-01

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

  18. Larvicidal, oviposition, and ovicidal effects of Artemisia annua (Asterales: Asteraceae) against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Cheah, Shao-Xiong; Tay, Jia-Wei; Chan, Lai-Keng; Jaal, Zairi

    2013-09-01

    This study focuses on the larvicidal, oviposition, and ovicidal effects of a crude extract of Artemisia annua against Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus. Dried cells of Artemisia annua from cell suspension cultures were extracted using hexane. The extract showed moderate larvicidal effects against mosquitoes. At 24-h post treatment, the LC50 values for Anopheles sinensis, Aedes aegypti, and Culex quinquefasciatus were recorded as 244.55, 276.14, and 374.99 ppm, respectively. The percentage mortality of larvae was directly proportional to the tested concentration. Anopheles sinensis was found to be the most susceptible species, whereas Culex quinquefasciatus was the most tolerant to the Artemisia annua extract. The results indicated that the Artemisia annua extract showed concentration-dependent oviposition deterrent activity and had a strong deterrent effect. At 500 ppm, the percentage effective repellency was more than 85% compared with the control group for all the species, with oviposition activity index values of -0.94, -0.95, and -0.78 for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. In the ovicidal assay, the percentage hatchability of eggs after treatment with 500 ppm of Artemisia annua extract was significantly lower than the control, with values of 48.84 ± 4.08, 38.42 ± 3.67, and 79.35 ± 2.09% for Aedes aegypti, Anopheles sinensis, and Culex quinquefasciatus, respectively. Artemisia annua was found to be more effective against Aedes aegypti and Anopheles sinensis compared with Culex quinquefasciatus. This study indicated that crude extract of A. annua could be a potential alternative for use in vector management programs.

  19. Acetylcholinesterase (Ace-1) target site mutation 119S is strongly diagnostic of carbamate and organophosphate resistance in Anopheles gambiae s.s. and Anopheles coluzzii across southern Ghana

    PubMed Central

    2013-01-01

    Background With high DDT resistance present throughout much of West Africa, carbamates and organophosphates are increasingly important alternatives to pyrethroids for indoor residual spraying (IRS). Though less widespread, resistance to both of these alternative insecticide classes has also been documented within the Anopheles gambiae species pair (formerly the M and S molecular forms) in West Africa. To manage insecticide efficacy, it is important to predict how and where resistance is likely to occur and spread, which could be aided by using molecular diagnostics with high predictive value. Methods Anopheles coluzzii and An. gambiae s.s. were collected from 18 sites throughout southern Ghana and bioassayed with bendiocarb, the most commonly applied carbamate, and an organophosphate, fenitrothion. The Ace-1 target site substitution G119S was genotyped by qPCR. Results Fenitrothion induced higher mortality than bendiocarb, though phenotypes correlated strongly across populations. Ace-1 119S was found at much higher frequency in An. gambiae s.s than An. coluzzii, exceeding 90% in a population from Greater Accra, the highest frequency reported to date. Ace-1 G119S was very strongly associated with resistance to both insecticides, providing high predictive power for diagnosis, though with some evidence for a differential effect between molecular forms for bendiocarb. Sequencing of the gene revealed a lack of variation in resistant alleles precluding determination of origin, but Ace-1 copy number variation was detected for the first time in Ghana. Conclusions The results validate G119S as a useful diagnostic of organophosphate and carbamate resistance within and among populations, whilst highlighting the potential for an aggregate nature of Ace-1 genotypes, which may comprise both single-copy and duplicated genes. Further work is now required to determine the distribution and resistance-association of Ace-1 duplication. PMID:24206629

  20. Cytogenetics of the Anopheles gambiae complex in Sudan, with special reference to An. arabiensis: relationships with East and West African populations.

    PubMed

    Petrarca, V; Nugud, A D; Ahmed, M A; Haridi, A M; Di Deco, M A; Coluzzi, M

    2000-06-01

    The species composition of malaria vector mosquitoes belonging to the Anopheles gambiae complex (Diptera: Culicidae) from >40 localities in Sudan, representing most ecological situations, was determined by analysis of ovarian polytene chromosomes. Of 2162 females, 93% were identified as An. arabiensis Patton and 7% were An. gambiae Giles sensu stricto. No hybrids were found between the two species. Anopheles arabiensis occurred in all but two sites, whereas An. gambiae s.s. was effectively limited to the southernmost, more humid localities. For chromosomal paracentric inversions, the degree of polymorphism was low in An. gambiae s.s. (inversions 2La, 2Rb and 2Rd), higher in An. arabiensis (inversions Xe, 2Ra, b, bc, d1, s; 3Ra, d). Anopheles gambiae samples from Sudan were all apparently panmictic, i.e. they did not show restricted gene flow such as observed among West African populations (interpreted as incipient speciation). Chromosomal inversion patterns of An. gambiae in southern Sudan showed characteristics of intergrading Savanna/Forest populations similar to those observed in comparable eco-climatic situations of West Africa. Anopheles arabiensis was polymorphic for inversion systems recorded in West Africa (2Ra, 2Rb, 2Rdl, 3Ra) and for a novel 2Rs polymorphism, overlapping with inversion systems 2Rb and 2Rd1. Samples carrying the 2Rs inversion were mostly from Khashm-el-Girba area in central-eastern Sudan. In the great majority of the samples all polymorphic inversions were found to be in Hardy-Weinberg equilibrium. Sudan populations of An. arabiensis should therefore be considered as generally panmictic. Anopheles arabiensis shows more inversion polymorphism in west than in east African populations. Sudan populations have more evident similarities with those from westwards than those from eastwards of the Great Rift Valley. The possible influence of the Rift on evolution of An. arabiensis is discussed.

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

    PubMed Central

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

    2011-01-01

    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 HgCl2 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 (<20% relative humidity), mosquitoes with reduced AgAQP1 protein survive significantly longer than controls. These studies support a role for AgAQP1 in water homeostasis during blood feeding and humidity adaptation of A. gambiae, a major mosquito vector of human malaria in sub-Saharan Africa. PMID:21444767

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

    PubMed

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

    2012-01-01

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

  3. Multicopper Oxidase-3 Is a Laccase Associated with the Peritrophic Matrix of Anopheles gambiae

    PubMed Central

    Lang, Minglin; Kanost, Michael R.; Gorman, Maureen J.

    2012-01-01

    The multicopper oxidase (MCO) family of enzymes includes laccases, which oxidize a broad range of substrates including polyphenols and phenylendiamines; ferroxidases, which oxidize ferrous iron; and several other oxidases with specific substrates such as ascorbate, bilirubin or copper. The genome of Anopheles gambiae, a species of mosquito, encodes five putative multicopper oxidases. Of these five, only AgMCO2 has known enzymatic and physiological functions: it is a highly conserved laccase that functions in cuticle pigmentation and tanning by oxidizing dopamine and dopamine derivatives. AgMCO3 is a mosquito-specific gene that is expressed predominantly in adult midguts and Malpighian tubules. To determine its enzymatic function, we purified recombinant AgMCO3 and analyzed its activity. AgMCO3 oxidized hydroquinone (a p-diphenol), the five o-diphenols tested, 2,2′-azino-bis(3-ethylbenzthiazoline-6-sulphonic acid) (ABTS), and p-phenylenediamine, but not ferrous iron. The catalytic efficiencies of AgMCO3 were similar to those of cuticular laccases (MCO2 orthologs), except that AgMCO3 oxidized all of the phenolic substrates with similar efficiencies whereas the MCO2 isoforms were less efficient at oxidizing catechol or dopa. These results demonstrate that AgMCO3 can be classified as a laccase and suggest that AgMCO3 has a somewhat broader substrate specificity than MCO2 orthologs. In addition, we observed AgMCO3 immunoreactivity in the peritrophic matrix, which functions as a selective barrier between the blood meal and midgut epithelial cells, protecting the midgut from mechanical damage, pathogens, and toxic molecules. We propose that AgMCO3 may oxidize toxic molecules in the blood meal leading to detoxification or to cross-linking of the molecules to the peritrophic matrix, thus targeting them for excretion. PMID:22479493

  4. Distribution of cuticular proteins in different structures of adult Anopheles gambiae.

    PubMed

    Zhou, Yihong; Badgett, Majors J; Bowen, John Hunter; Vannini, Laura; Orlando, Ron; Willis, Judith H

    2016-08-01

    Anopheles gambiae devotes over 2% (295) of its protein coding genes to structural cuticular proteins (CPs) that have been classified into 13 different families plus ten low complexity proteins not assigned to families. Small groups of genes code for identical proteins reducing the total number of unique cuticular proteins to 282. Is the large number because different structures utilize different CPs, or are all of the genes widely expressed? We used LC-MS/MS to learn how many products of these genes were found in five adult structures: Johnston's organs, the remainder of the male antennae, eye lenses, legs, and wings. Data were analyzed against both the entire proteome and a smaller database of just CPs. We recovered unique peptides for 97 CPs and shared peptides for another 35. Members of 11 of the 13 families were recovered as well as some unclassified. Only 11 CPs were present exclusively in only one structure while 43 CPs were recovered from all five structures. A quantitative analysis, using normalized spectral counts, revealed that only a few CPs were abundant in each structure. When the MS/MS data were run against the entire proteome, the majority of the top hits were to CPs, but peptides were recovered from an additional 467 proteins. CP peptides were frequently recovered from chitin-binding domains, confirming that protein-chitin interactions are not mediated by covalent bonds. Comparison with three other MS/MS analyses of cuticles or cuticle-rich structures augmented the current analysis. Our findings provide new insights into the composition of different mosquito structures and reveal the complexity of selection and utilization of genes coding for structural cuticular proteins.

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

    PubMed Central

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

    2012-01-01

    The association between fitness-related phenotypic traits and an environmental gradient offers one of the best opportunities to study the interplay between natural selection and migration. In cases in which specific genetic variants also show such clinal patterns, it may be possible to uncover the mutations responsible for local adaptation. The malaria vector, Anopheles gambiae, is associated with a latitudinal cline in aridity in Cameroon; a large inversion on chromosome 2L of this mosquito shows large differences in frequency along this cline, with high frequencies of the inverted karyotype present in northern, more arid populations and an almost complete absence of the inverted arrangement in southern populations. Here we use a genome resequencing approach to investigate patterns of population divergence along the cline. By sequencing pools of individuals from both ends of the cline as well as in the center of the cline—where the inversion is present in intermediate frequency—we demonstrate almost complete panmixia across collinear parts of the genome and high levels of differentiation in inverted parts of the genome. Sequencing of separate pools of each inversion arrangement in the center of the cline reveals large amounts of gene flux (i.e., gene conversion and double crossovers) even within inverted regions, especially away from the inversion breakpoints. The interplay between natural selection, migration, and gene flux allows us to identify several candidate genes responsible for the match between inversion frequency and environmental variables. These results, coupled with similar conclusions from studies of clinal variation in Drosophila, point to a number of important biological functions associated with local environmental adaptation. PMID:22209907

  6. Mosquito larvicidal activity of seaweeds extracts against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus

    PubMed Central

    Ali, Mohamed Yacoob Syed; Ravikumar, Sundaram; Beula, Johanson Margaret

    2013-01-01

    Objective To identify the larvicidal activity of the seaweed extracts against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus Methods Seaweed extracts of Ulva lactuca, Caulerpa racemosa (C. racemosa), Sargassum microystum, Caulerpa scalpelliformis, Gracilaria corticata, Turbinaria decurrens, Turbinaria conoides and Caulerpa toxifolia were dissolved in DMSO to prepare a graded series of concentration. The test for the larvicidal effect of seaweeds against mosquitos larvae was conducted in accordance with the WHO standard method. Batches of 25 early 4th instar larvae of three mosquitoes were transferred to 250 mL enamel bowl containing 199 mL of distilled water and 1 mL of plant extracts (10-100 µg). Each experiment was conducted with triplicate with concurrent a control group. Results Among the seaweeds extract, C. racemosa showed toxicity against 4th instar larvae of Aedes aegypti, Culex quinquefasciatus, Anopheles stephensi with equivalent LC50 value (0.055 6±0.010 3) µg/mL, (0.067 5±0.136 0) µg/mL and (0.066 1±0.007 6) µg/mL, respectively. Conclusions The present study concluded that, the mosquito larvicidal property of C. racemosa might be the prospective alternative source to control the mosquitoes.

  7. Screening of Methanolic Plant Extracts against Larvae of Aedes aegypti and Anopheles stephensi in Mysore

    PubMed Central

    Mohankumar, Thirumalapura Krishnaiah; Shivanna, Kumuda Sathigal; Achuttan, Vijayan Valiakottukal

    2016-01-01

    Background: Mosquitoes transmit serious human diseases, causing millions of death every year. Vector control is facing a threat due to the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. Nine different locally available medicinally important plants suspected to posse larvicidal property were screened against fourth instar larvae of Aedes aegypti and Anopheles stephensi to a series of concentrations of the methanolic extracts. Methods: Susceptibility tests on Ae. aegypti and An. stephensi were conducted using standard WHO methods. The larvae of two mosquito species were exposed to methanolic extracts and mortality counts were made after 24 hours of exposure as per WHO method. Larvae of Ae. aegypti were more susceptible than that of An. stephensi. Results: Among the nine plant species tested, Annona reticulata leaf extract was more effective against Ae. aegypti larvae with LC50 and LC90 values of 95.24 and 262.64 ppm respectively and against An. stephensi larvae 262.71 and 636.94 ppm respectively. The least efficacy was in Cosmos bipinnatus with LC50 and LC90 values of 442.6 and 1225.93 ppm against Ae. aegypti and LC50 and LC90 values of 840.69 and 1334.01 ppm of Thespesia populnea against An. stephensi. Conclusion: The crude methanolic extract of the An. reticulata with good larvicidal efficacy could be considered for further characterization to control mosquito vectors instead of chemical insecticides. High efficacy found in An. reticulata extract will be considered for further studies to isolate the bioactive compound. PMID:27308289

  8. Melanotic Pathology and Vertical Transmission of the Gut Commensal Elizabethkingia meningoseptica in the Major Malaria Vector Anopheles gambiae

    PubMed Central

    Christophides, Georges K.

    2013-01-01

    Background The resident gut flora is known to have significant impacts on the life history of the host organism. Endosymbiotic bacterial species in the Anopheles mosquito gut are potent modulators of sexual development of the malaria parasite, Plasmodium, and thus proposed as potential control agents of malaria transmission. Results Here we report a melanotic pathology in the major African malaria vector Anopheles gambiae, caused by the dominant mosquito endosymbiont Elizabethkingiameningoseptica. Transfer of melanised tissues into the haemolymph of healthy adult mosquitoes or direct haemolymph inoculation with isolated E. meningoseptica bacteria were the only means for transmission and de novo formation of melanotic lesions, specifically in the fat body tissues of recipient individuals. We show that E. meningoseptica can be vertically transmitted from eggs to larvae and that E. meningoseptica-mono-associated mosquitoes display significant mortality, which is further enhanced upon Plasmodium infection, suggesting a synergistic impact of E. meningoseptica and Plasmodium on mosquito survival. Conclusion The high pathogenicity and permanent association of E. meningoseptica with An. Gambiae through vertical transmission constitute attractive characteristics towards the potential design of novel mosquito/malaria biocontrol strategies. PMID:24098592

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

    PubMed Central

    2010-01-01

    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. However, traps baited with

  10. Comparison of the oxidative phosphorylation (OXPHOS) nuclear genes in the genomes of Drosophila melanogaster, Drosophila pseudoobscura and Anopheles gambiae

    PubMed Central

    Tripoli, Gaetano; D'Elia, Domenica; Barsanti, Paolo; Caggese, Corrado

    2005-01-01

    Background In eukaryotic cells, oxidative phosphorylation (OXPHOS) uses the products of both nuclear and mitochondrial genes to generate cellular ATP. Interspecies comparative analysis of these genes, which appear to be under strong functional constraints, may shed light on the evolutionary mechanisms that act on a set of genes correlated by function and subcellular localization of their products. Results We have identified and annotated the Drosophila melanogaster, D. pseudoobscura and Anopheles gambiae orthologs of 78 nuclear genes encoding mitochondrial proteins involved in oxidative phosphorylation by a comparative analysis of their genomic sequences and organization. We have also identified 47 genes in these three dipteran species each of which shares significant sequence homology with one of the above-mentioned OXPHOS orthologs, and which are likely to have originated by duplication during evolution. Gene structure and intron length are essentially conserved in the three species, although gain or loss of introns is common in A. gambiae. In most tissues of D. melanogaster and A. gambiae the expression level of the duplicate gene is much lower than that of the original gene, and in D. melanogaster at least, its expression is almost always strongly testis-biased, in contrast to the soma-biased expression of the parent gene. Conclusions Quickly achieving an expression pattern different from the parent genes may be required for new OXPHOS gene duplicates to be maintained in the genome. This may be a general evolutionary mechanism for originating phenotypic changes that could lead to species differentiation. PMID:15693940

  11. Repellent, Irritant and Toxic Effects of 20 Plant Extracts on Adults of the Malaria Vector Anopheles gambiae Mosquito

    PubMed Central

    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. PMID:24376515

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

    PubMed

    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.

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

    PubMed

    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. PMID:24376515

  14. Large fluctuations in the effective population size of the malaria mosquito Anopheles gambiae s.s. during vector control cycle.

    PubMed

    Hodges, Theresa K; Athrey, Giridhar; Deitz, Kevin C; Overgaard, Hans J; Matias, Abrahan; Caccone, Adalgisa; Slotman, Michel A

    2013-12-01

    On Bioko Island, Equatorial Guinea, indoor residual spraying (IRS) has been part of the Bioko Island Malaria Control Project since early 2004. Despite success in reducing childhood infections, areas of high transmission remain on the island. We therefore examined fluctuations in the effective population size (N e ) of the malaria vector Anopheles gambiae in an area of persistent high transmission over two spray rounds. We analyzed data for 13 microsatellite loci from 791 An. gambiae specimens collected at six time points in 2009 and 2010 and reconstructed the demographic history of the population during this period using approximate Bayesian computation (ABC). Our analysis shows that IRS rounds have a large impact on N e , reducing it by 65%-92% from prespray round N e . More importantly, our analysis shows that after 3-5 months, the An. gambiae population rebounded by 2818% compared shortly following the spray round. Our study underscores the importance of adequate spray round frequency to provide continuous suppression of mosquito populations and that increased spray round frequency should substantially improve the efficacy of IRS campaigns. It also demonstrates the ability of ABC to reconstruct a detailed demographic history across only a few tens of generations in a large population.

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

    PubMed

    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.

  16. Large fluctuations in the effective population size of the malaria mosquito Anopheles gambiae s.s. during vector control cycle

    PubMed Central

    Hodges, Theresa K; Athrey, Giridhar; Deitz, Kevin C; Overgaard, Hans J; Matias, Abrahan; Caccone, Adalgisa; Slotman, Michel A

    2013-01-01

    On Bioko Island, Equatorial Guinea, indoor residual spraying (IRS) has been part of the Bioko Island Malaria Control Project since early 2004. Despite success in reducing childhood infections, areas of high transmission remain on the island. We therefore examined fluctuations in the effective population size (Ne) of the malaria vector Anopheles gambiae in an area of persistent high transmission over two spray rounds. We analyzed data for 13 microsatellite loci from 791 An. gambiae specimens collected at six time points in 2009 and 2010 and reconstructed the demographic history of the population during this period using approximate Bayesian computation (ABC). Our analysis shows that IRS rounds have a large impact on Ne, reducing it by 65%–92% from prespray round Ne. More importantly, our analysis shows that after 3–5 months, the An. gambiae population rebounded by 2818% compared shortly following the spray round. Our study underscores the importance of adequate spray round frequency to provide continuous suppression of mosquito populations and that increased spray round frequency should substantially improve the efficacy of IRS campaigns. It also demonstrates the ability of ABC to reconstruct a detailed demographic history across only a few tens of generations in a large population. PMID:24478799

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

    PubMed Central

    2014-01-01

    Background 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 behaviour especially on how the extracts of such plants impact on the fitness of this vector has not been explored. This study determined the toxicity and larvicidal activity on this vector of extracts from six selected plants found in Kenya and two compounds identified from Ricinus communis: 3-carbonitrile-4-methoxy-N-methyl-2-pyridone (ricinine), and its carboxylic acid derivative 3-carboxy-4-methoxy-N-methyl-2-pyridone, the latter compound being reported for the first time from this plant. Methods Feeding assays tested for toxic effects of extracts from the plants Artemisia afra Jacq. ex Willd, Bidens pilosa L., Parthenium hysterophorus L., Ricinus coummunis L., Senna didymobotrya Fresen. and Tithonia diversifolia Hemsl. on adult females and larvicidal activity was tested against third-instar larvae of Anopheles gambiae s.s. Mortality of larvae and adult females was monitored for three and eight days, respectively; Probit analysis was used to calculate LC50. Survival was analysed with Kaplan-Meier Model. LC-MS was used to identify the pure compounds. Results Of the six plants screened, extracts from T. diversifolia and R. communis were the most toxic against adult female mosquitoes after 7 days of feeding, with LC50 of 1.52 and 2.56 mg/mL respectively. Larvicidal activity of all the extracts increased with the exposure time with the highest mortality recorded for the extract from R. communis after 72 h of exposure (LC50 0.18 mg/mL). Mosquitoes fed on solutions of the pure compounds, 3-carboxy-4-methoxy-N-methyl-2-pyridone and ricinine survived almost as long as those fed on the R. communis extract with mean survival of 4.93 ± 0.07, 4.85 ± 0.07 and 4.50 ± 0.05 days respectively. Conclusions Overall, these findings demonstrate that extracts from the six plant species exhibit

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Objective: To determine the larvicidal activity of hexane, chloroform, ethyl acetate, acetone, and methanol extracts of Orthosiphon thymiflorus leaves against Anopheles stephensi, Culex quinquefasciatus and Aedes aegypti. Methods: Larvicidal activity was determined in laboratory bioassays using var...

  19. Preliminary lack of evidence for simian odour preferences of savanna populations of Anopheles gambiae and other malaria vectors.

    PubMed

    Costantini, C; Diallo, M

    2001-12-01

    The behavioural response to several culicine and anopheline mosquitoes to the odour of alternative hosts (human vs monkey) arranged in a choice set-up using odour-baited entry traps (OBETs) was assessed in a field experiment in south-eastern Senegal. The experimental protocol followed procedures analogous to those adopted in olfactometer laboratory tests. Two adult Cercopithecus aethiops and a child of similar mass slept inside separate tents and their odours were drawn to each one of two paired OBETs so that approaching mosquitoes could experience both odour-laden streams before "choosing" to fly against one of the two air currents and into the trap. The traps were set up in a riverine forest clearing near the town of Kedougou, where primates (Papio papio, Cercopithecus aethiops, and Erythrocebus patas) are common. A total of 192 mosquitoes belonging to 4 genera was captured during 8 trap nights. All major human malaria vectors including Anopheles gambiae sensu lato, An. funestus, and An. nili, which constituted the bulk of the trap catch (N = 153), clearly expressed a preference for human odour, with > 90% of captured mosquitoes caught in the human-baited trap. A sub-sample of specimens belonging to the An. gambiae complex caught in both traps was identified by rDNA-PCR and RFLP as An. gambiae sensu stricto molecular form S (7/10), and An. arabiensis (3/10). The only species that did not show a preference for the alternative odour-laden air streams, among those caught in significant numbers, were mosquitoes of the genus Mansonia, with both Ma. uniformis and Ma. africana weakly preferring human odour, but not at a statistically significant level. These results are in accordance with the hypothesis that the strongly anthropophilic feeding preferences of An. gambiae did not evolve from an ancestral association with non-human primates.

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

    PubMed Central

    Obbard, Darren J; Welch, John J; Little, Tom J

    2009-01-01

    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. PMID:19497100

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

    PubMed

    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.

  2. Gambia.

    PubMed

    1985-05-01

    This compendium on the Gambia contains a summary of 1) current and projected demographic indicators, 2) government population policies and concerns, and 3) the current status of the national population data collecting system. The information is further summarized in a 1-page fact sheet. Gambia is the smallest African nation and has an average density of 50 persons/square kilometer. The total population in 1980 was 583,000. Between 1975-80 the annual population growth rate was 2.2%, and the annual rate of natural increase was 1.8%. Preliminary results of the 1983 census indicate that the annual growth rate between 1973-83 was 2.5%. The middle range, projected total population for 2000 is 898,000. The Gambia's recent population growth is attributed to a decline in mortality and a relatively high level of uncontrolled immigration from surrounding countries. The emigration rate was relatively low. Life expectancy at birth was 33.5 years, and in 1975-80, the infant mortality rate was 204. During 1975-80, the crude birth rate was 48.3, and the net reproduction rate was 1.7. The urban rate was 18.5% (1980), and the annual urban growth rate was 4.4% (1975-80). Per capita gross domestic product was US$369 (1979), and in 1983, 76.7% of the labor force was engaged in agriculture. The Gambia's second 5-year plan (1981-82 to 1985-86) contains the nation's first formal statement of population policy. The plan calls for controlling population growth by reducing mortality, fertility, and immigration and recognizes the relationship between economic development and population. The government recognizes the difficulties involved in providing basic services under conditions of rapid population growth and that rapid population is contributing to the deterioration of the environment. The country's primary development goals are to diversify the economy and increase productivity, especially in the agricultural sector. A reduction in mortality will be achieved by promoting preventive and

  3. The CPCFC cuticular protein family: Anatomical and cuticular locations in Anopheles gambiae and distribution throughout Pancrustacea.

    PubMed

    Vannini, Laura; Bowen, John Hunter; Reed, Tyler W; Willis, Judith H

    2015-10-01

    Arthropod cuticles have, in addition to chitin, many structural proteins belonging to diverse families. Information is sparse about how these different cuticular proteins contribute to the cuticle. Most cuticular proteins lack cysteine with the exception of two families (CPAP1 and CPAP3), recently described, and the one other that we now report on that has a motif of 16 amino acids first identified in a protein, Bc-NCP1, from the cuticle of nymphs of the cockroach, Blaberus craniifer (Jensen et al., 1997). This motif turns out to be present as two or three copies in one or two proteins in species from many orders of Hexapoda. We have named the family of cuticular proteins with this motif CPCFC, based on its unique feature of having two cysteines interrupted by five amino acids (C-X(5)-C). Analysis of the single member of the family in Anopheles gambiae (AgamCPCFC1) revealed that its mRNA is most abundant immediately following ecdysis in larvae, pupae and adults. The mRNA is localized primarily in epidermis that secretes hard cuticle, sclerites, setae, head capsules, appendages and spermatheca. EM immunolocalization revealed the presence of the protein, generally in endocuticle of legs and antennae. A phylogenetic analysis found proteins bearing this motif in 14 orders of Hexapoda, but not in some species for which there are complete genomic data. Proteins were much longer in Coleoptera and Diptera than in other orders. In contrast to the 1 and occasionally 2 copies in other species, a dragonfly, Ladona fulva, has at least 14 genes coding for family members. CPCFC proteins were present in four classes of Crustacea with 5 repeats in one species, and motifs that ended C-X(7)-C in Malacostraca. They were not detected, except as obvious contaminants, in any other arthropod subphyla or in any other phylum. The conservation of CPCFC proteins throughout the Pancrustacea and the small number of copies in individual species indicate that, when present, these proteins are

  4. Insecticide resistance mechanisms associated with different environments in the malaria vector Anopheles gambiae: a case study in Tanzania

    PubMed Central

    2014-01-01

    Background Resistance of mosquitoes to insecticides is a growing concern in Africa. Since only a few insecticides are used for public health and limited development of new molecules is expected in the next decade, maintaining the efficacy of control programmes mostly relies on resistance management strategies. Developing such strategies requires a deep understanding of factors influencing resistance together with characterizing the mechanisms involved. Among factors likely to influence insecticide resistance in mosquitoes, agriculture and urbanization have been implicated but rarely studied in detail. The present study aimed at comparing insecticide resistance levels and associated mechanisms across multiple Anopheles gambiae sensu lato populations from different environments. Methods Nine populations were sampled in three areas of Tanzania showing contrasting agriculture activity, urbanization and usage of insecticides for vector control. Insecticide resistance levels were measured in larvae and adults through bioassays with deltamethrin, DDT and bendiocarb. The distribution of An. gambiae sub-species and pyrethroid target-site mutations (kdr) were investigated using molecular assays. A microarray approach was used for identifying transcription level variations associated to different environments and insecticide resistance. Results Elevated resistance levels to deltamethrin and DDT were identified in agriculture and urban areas as compared to the susceptible strain Kisumu. A significant correlation was found between adult deltamethrin resistance and agriculture activity. The subspecies Anopheles arabiensis was predominant with only few An. gambiae sensu stricto identified in the urban area of Dar es Salaam. The L1014S kdr mutation was detected at elevated frequency in An gambiae s.s. in the urban area but remains sporadic in An. arabiensis specimens. Microarrays identified 416 transcripts differentially expressed in any area versus the susceptible reference

  5. The MitoDrome database annotates and compares the OXPHOS nuclear genes of Drosophila melanogaster, Drosophila pseudoobscura and Anopheles gambiae.

    PubMed

    D'Elia, Domenica; Catalano, Domenico; Licciulli, Flavio; Turi, Antonio; Tripoli, Gaetano; Porcelli, Damiano; Saccone, Cecilia; Caggese, Corrado

    2006-10-01

    The oxidative phosphorylation (OXPHOS) is the primary energy-producing process of all aerobic organisms and the only cellular function under the dual control of both the mitochondrial and the nuclear genomes. Functional characterization and evolutionary study of the OXPHOS system is of great importance for the understanding of many as yet unclear aspects of nucleus-mitochondrion genomic co-evolution and co-regulation gene networks. The MitoDrome database is a web-based database which provides genomic annotations about nuclear genes of Drosophila melanogaster encoding for mitochondrial proteins. Recently, MitoDrome has included a new section annotating genomic information about OXPHOS genes in Drosophila pseudoobscura and Anopheles gambiae and their comparative analysis with their Drosophila melanogaster and human counterparts. The introduction of this new comparative annotation section into MitoDrome is expected to be a useful resource for both functional and structural genomics related to the OXPHOS system.

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

    PubMed Central

    2012-01-01

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

  7. Diel timing and frequency of sugar feeding in the mosquito Anopheles gambiae, depending on sex, gonotrophic state and resource availability.

    PubMed

    Gary, R E; Foster, W A

    2006-09-01

    Little is known about the sugar-feeding behaviour of equatorial Africa's principal vector of malaria, Anopheles gambiae Giles (Diptera: Culicidae). It is suspected to feed on plant sugar infrequently, but possibly the timing depends on environmental circumstances, and males may differ markedly from females. These points of uncertainty were clarified in the laboratory, by monitoring both diel and longterm sugar-feeding activity in both sexes. Males fed on sugar in a nocturnal diel rhythm closely approximating non-specific flight activity. Female diel sugar-feeding patterns resembled published rhythms and cycles of host seeking. Males sugar fed nightly at an average frequency of about twice per night, sustained over 17 days. This was substantially higher than the sugar-feeding frequency of females that were allowed both blood and oviposition sites every night: they averaged about one sugar feed in every 4 nights. These females fed on sugar between gonotrophic cycles, after eggs were mature but before the next bloodmeal. They did not sugar feed during the 2 days after blood feeding, while blood was being digested and the eggs developed. A slight delay in the availability of either the oviposition site or blood led to an increase in female sugar-feeding frequency: they averaged more than once per night until the delayed resource was made available. These observations support the conclusion that sugar feeding is a normal part of the biology of both sexes of An. gambiae.

  8. Effect of temperature on the development of the aquatic stages of Anopheles gambiae sensu stricto (Diptera: Culicidae).

    PubMed

    Bayoh, M N; Lindsay, S W

    2003-10-01

    Global warming may affect the future pattern of many arthropod-borne diseases, yet the relationship between temperature and development has been poorly described for many key vectors. Here the development of the aquatic stages of Africa's principal malaria vector, Anopheles gambiae s.s. Giles, is described at different temperatures. Development time from egg to adult was measured under laboratory conditions at constant temperatures between 10 and 40 degrees C. Rate of development from one immature stage to the next increased at higher temperatures to a peak around 28 degrees C and then declined. Adult development rate was greatest between 28 and 32 degrees C, although adult emergence was highest between 22 and 26 degrees C. No adults emerged below 18 degrees C or above 34 degrees C. Non-linear models were used to describe the relationship between developmental rate and temperature, which could be used for developing process-based models of malaria transmission. The utility of these findings is demonstrated by showing that a map where the climate is suitable for the development of aquatic stages of A. gambiae s.s. corresponded closely with the best map of malaria risk currently available for Africa.

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

    PubMed

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

    2016-01-01

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

  10. Variation in susceptibility of African Plasmodium falciparum malaria parasites to TEP1 mediated killing in Anopheles gambiae mosquitoes

    PubMed Central

    Eldering, Maarten; Morlais, Isabelle; van Gemert, Geert-Jan; van de Vegte-Bolmer, Marga; Graumans, Wouter; Siebelink-Stoter, Rianne; Vos, Martijn; Abate, Luc; Roeffen, Will; Bousema, Teun; Levashina, Elena A.; Sauerwein, Robert W.

    2016-01-01

    Anopheles gambiae s.s. mosquitoes are efficient vectors for Plasmodium falciparum, although variation exists in their susceptibility to infection. This variation depends partly on the thioester-containing protein 1 (TEP1) and TEP depletion results in significantly elevated numbers of oocysts in susceptible and resistant mosquitoes. Polymorphism in the Plasmodium gene coding for the surface protein Pfs47 modulates resistance of some parasite laboratory strains to TEP1-mediated killing. Here, we examined resistance of P. falciparum isolates of African origin (NF54, NF165 and NF166) to TEP1-mediated killing in a susceptible Ngousso and a refractory L3–5 strain of A. gambiae. All parasite clones successfully developed in susceptible mosquitoes with limited evidence for an impact of TEP1 on transmission efficiency. In contrast, NF166 and NF165 oocyst densities were strongly reduced in refractory mosquitoes and TEP1 silencing significantly increased oocyst densities. Our results reveal differences between African P. falciparum strains in their capacity to evade TEP1-mediated killing in resistant mosquitoes. There was no significant correlation between Pfs47 genotype and resistance of a given P. falciparum isolate for TEP1 killing. These data suggest that polymorphisms in this locus are not the sole mediators of immune evasion of African malaria parasites. PMID:26861587

  11. Molecular and functional characterization of Anopheles gambiae inward rectifier potassium (Kir1) channels: A novel role in egg production

    PubMed Central

    Raphemot, Rene; Estévez-Lao, Tania Y.; Rouhier, Matthew F.; Piermarini, Peter M.; Denton, Jerod S.; Hillyer, Julián F.

    2014-01-01

    Inward rectifier potassium (Kir) channels play essential roles in regulating diverse physiological processes. Although Kir channels are encoded in mosquito genomes, their functions remain largely unknown. In this study, we identified the members of the Anopheles gambiae Kir gene family and began to investigate their function. Notably, we sequenced the A. gambiae Kir1 (AgKir1) gene and showed that it encodes all the canonical RIP features of a Kir channel: an ion pore that is composed of a pore helix and a selectivity filter, two transmembrane domains that flank the ion pore, and the so-called G-loop. Heterologous expression of AgKir1 in Xenopus oocytes revealed that this gene encodes a functional, barium-sensitive Kir channel. Quantitative RT-PCR experiments then showed that relative AgKir1 mRNA levels are highest in the pupal stage, and that AgKir1 mRNA is enriched in the adult ovaries. Gene silencing of AgKir1 by RNA interference did not affect the survival of female mosquitoes following a blood in mosquito fecundity, and further validates them as promising molecular targets for the meal, but decreased their egg output. These data provide evidence for a new role of Kir channels development of a new class of mosquitocides to be used in vector control. PMID:24855023

  12. Differential attractiveness of human foot odours to Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) and variation in their chemical composition.

    PubMed

    Omolo, M O; Njiru, B; Ndiege, I O; Musau, R M; Hassanali, A

    2013-10-01

    Counter flow geometry (CFG) traps (American Biophysics) baited with foot odours (adsorbed overnight on a combination of a nylon and a cotton socks) from 4 groups of 4 male volunteers were initially used in a screen-house to compare and grade the attractiveness of Anopheles gambiae Giles sensu stricto to these odours. Ten individuals were then selected from the 4 groups for further grading in which mosquito attractiveness to odours adsorbed on socks worn by each of the individuals was compared against a control (clean, un-worn cotton and nylon socks). A gradation of attractiveness was found, with the most attractive foot odour catching 8-fold more mosquitoes than the least attractive one (t-test, p=0.001). For comparison of the chromatographic profiles of the foot odours and identification of their constituents, six adsorbents (tenax, chromsorb, porapak Q, activated charcoal, reverse phase octadecyl and octyl bonded silica) were evaluated for their suitability in trapping the volatiles in a static mode. Of these, porapak Q was found to be the most effective. Comparison of the gas-chromatographic (GC) and gas chromatography-linked mass spectrometry (GC-MS) profiles of blends collected from the most and least attractive feet on porapak Q adsorbent revealed both qualitative and quantitative differences. The implication of our finding on differential attraction of An. gambiae to their preferred feeding site of different human subjects is highlighted.

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

    PubMed

    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

    2014-01-01

    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. PMID:25525826

  14. Genetic differentiation in the African malaria vector, Anopheles gambiae s.s., and the problem of taxonomic status.

    PubMed Central

    Gentile, Gabriele; Della Torre, Alessandra; Maegga, Bertha; Powell, Jeffrey R; Caccone, Adalgisa

    2002-01-01

    Of the seven recognized species of the Anopheles gambiae complex, A. gambiae s.s. is the most widespread and most important vector of malaria. It is becoming clear that, in parts of West Africa, this nominal species is not a single panmictic unit. We found that the internal transcribed spacer (ITS) of the X-linked rDNA has two distinct sequences with three fixed nucleotide differences; we detected no heterozygotes at these three sites, even in areas of sympatry of the two ITS types. The intergenic spacer (IGS) of this region also displays two distinct sequences that are in almost complete linkage disequilibrium with the distinct ITS alleles. We have designated these two types as S/type I and M/type II. These rDNA types correspond at least partly to the previously recognized chromosomal forms. Here we expand the geographic range of sampling to 251 individuals from 38 populations. Outside of West Africa, a single rDNA type, S/type I, corresponds to the Savanna chromosomal form. In West Africa, both types are often found in a single local sample. To understand if these findings might be due to unusual behavior of the rDNA region, we sequenced the same region for 46 A. arabiensis, a sympatric sibling species. No such distinct discontinuity was observed for this species. Autosomal inversions in one chromosome arm (2R), an insecticide resistance gene on 2L, and this single X-linked region indicate at least two genetically differentiated subpopulations of A. gambiae. Yet, rather extensive studies of other regions of the genome have failed to reveal genetic discontinuity. Evidently, incomplete genetic isolation exists within this single nominal species. PMID:12196401

  15. How the Malaria Vector Anopheles gambiae Adapts to the Use of Insecticide-Treated Nets by African Populations

    PubMed Central

    Ndiath, Mamadou Ousmane; Mazenot, Catherine; Sokhna, Cheikh; Trape, Jean-François

    2014-01-01

    Background Insecticide treated bed nets have been recommended and proven efficient as a measure to protect African populations from malaria mosquito vector Anopheles spp. This study evaluates the consequences of bed nets use on vectors resistance to insecticides, their feeding behavior and malaria transmission in Dielmo village, Senegal, were LLINs were offered to all villagers in July 2008. Methods Adult mosquitoes were collected monthly from January 2006 to December 2011 by human landing catches (HLC) and by pyrethroid spray catches (PCS). A randomly selected sub-sample of 15–20% of An. gambiae s.l. collected each month was used to investigate the molecular forms of the An. gambiae complex, kdr mutations, and Plasmodium falciparum circumsporozoite (CSP) rate. Malaria prevalence and gametocytaemia in Dielmo villagers were measured quarterly. Results Insecticide susceptible mosquitoes (wild kdr genotype) presented a reduced lifespan after LLINs implementation but they rapidly adapted their feeding behavior, becoming more exophageous and zoophilic, and biting earlier during the night. In the meantime, insecticide-resistant specimens (kdr L1014F genotype) increased in frequency in the population, with an unchanged lifespan and feeding behaviour. P. falciparum prevalence and gametocyte rate in villagers decreased dramatically after LLINs deployment. Malaria infection rate tended to zero in susceptible mosquitoes whereas the infection rate increased markedly in the kdr homozygote mosquitoes. Conclusion Dramatic changes in vector populations and their behavior occurred after the deployment of LLINs due to the extraordinary adaptative skills of An. gambiae s. l. mosquitoes. However, despite the increasing proportion of insecticide resistant mosquitoes and their almost exclusive responsibility in malaria transmission, the P. falciparum gametocyte reservoir continued to decrease three years after the deployment of LLINs. PMID:24892677

  16. Ovicidal and repellent activities of botanical extracts against Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi (Diptera: Culicidae)

    PubMed Central

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

    2011-01-01

    Objective To determine the ovicidal and repellent activities of methanol leaf extract of Ervatamia coronaria (E. coronaria) and Caesalpinia pulcherrima (C. pulcherrima) against Culex quinquefasciatus (Cx. quinquefasciatus), Aedes aegypti (Ae. aegypti) and Anopheles stephensi (An. stephensi). Methods The ovicidal activity was determined against three mosquito species at various concentrations ranging from 50-450 ppm under the laboratory conditions. The hatch rates were assessed 48 h after treatment. The repellent efficacy was determined against three mosquito species at three concentrations viz., 1.0, 2.5 and 5.0 mg/cm2 under the laboratory conditions. Results The crude extract of E. coronaria exerted zero hatchability (100% mortality) at 250, 200 and 150 ppm for Cx. quinquefasciatus, Ae. aegypti and An. stephensi, respectively. The crude extract of C. pulcherrima exerted zero hatchability (100% mortality) at 375, 300 and 225 ppm for Cx. quinquefasciatus, Ae. aegypti and An. Stephensi, respectively. The methanol extract of E. coronaria found to be more repellenct than C. pulcherrima extract. A higher concentration of 5.0 mg/cm2 provided 100% protection up to 150, 180 and 210 min against Cx. quinquefasciatus, Ae. aegypti and An. stephensi, respectively. The results clearly showed that repellent activity was dose dependent. Conclusions From the results it can be concluded the crude extracts of E. coronaria and C. pulcherrima are an excellent potential for controlling Cx. quinquefasciatus, Ae. aegypti and An. stephensi mosquitoes. PMID:23569723

  17. Organ-Specific Splice Variants of Aquaporin Water Channel AgAQP1 in the Malaria Vector Anopheles gambiae

    PubMed Central

    Tsujimoto, Hitoshi; Liu, Kun; Linser, Paul J.; Agre, Peter; Rasgon, Jason L.

    2013-01-01

    Background Aquaporin (AQP) water channels are important for water homeostasis in all organisms. Malaria transmission is dependent on Anopheles mosquitoes. Water balance is a major factor influencing mosquito survival, which may indirectly affect pathogen transmission. Methodology/Principal Findings We obtained full-length mRNA sequences for Anopheles gambiae aquaporin 1 (AgAQP1) and identified two splice variants for the gene. In vitro expression analysis showed that both variants transported water and were inhibited by Hg2+. One splice variant (AgAQP1A) was exclusively expressed in adult female ovaries indicating a function in mosquito reproduction. The other splice variant (AgAQP1B) was expressed in the midgut, malpighian tubules and the head in adult mosquitoes. Immunolabeling showed that in malpighian tubules, AgAQP1 is expressed in principal cells in the proximal portion and in stellate cells in the distal portion. Moreover, AgAQP1 is expressed in Johnston’s organ (the “ear”), which is important for courtship behavior. Conclusions And Significance These results suggest that AgAQP1 may play roles associated with mating (courtship) and reproduction in addition to water homeostasis in this important African malaria vector. PMID:24066188

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

    PubMed Central

    2011-01-01

    Background The mosquito, Anopheles gambiae, is the primary vector of human malaria, a disease responsible for millions of deaths each year. To improve strategies for controlling transmission of the causative parasite, Plasmodium falciparum, we require a thorough understanding of the developmental mechanisms, physiological processes and evolutionary pressures affecting life-history traits in the mosquito. Identifying genes expressed in particular tissues or involved in specific biological processes is an essential part of this process. Results In this study, we present transcription profiles for ~82% of annotated Anopheles genes in dissected adult male and female tissues. The sensitivity afforded by examining dissected tissues found gene activity in an additional 20% of the genome that is undetected when using whole-animal samples. The somatic and reproductive tissues we examined each displayed patterns of sexually dimorphic and tissue-specific expression. By comparing expression profiles with Drosophila melanogaster we also assessed which genes are well conserved within the Diptera versus those that are more recently evolved. Conclusions Our expression atlas and associated publicly available database, the MozAtlas (http://www.tissue-atlas.org), provides information on the relative strength and specificity of gene expression in several somatic and reproductive tissues, isolated from a single strain grown under uniform conditions. The data will serve as a reference for other mosquito researchers by providing a simple method for identifying where genes are expressed in the adult, however, in addition our resource will also provide insights into the evolutionary diversity associated with gene expression levels among species. PMID:21649883

  19. Pyriproxyfen for mosquito control: female sterilization or horizontal transfer to oviposition substrates by Anopheles gambiae sensu stricto and Culex quinquefasciatus

    PubMed Central

    2014-01-01

    Background The use of gravid mosquitoes as vehicles to auto-disseminate larvicides was recently demonstrated for the transfer of pyriproxyfen (PPF) by container-breeding Aedes mosquitoes and presents an appealing idea to explore for other disease vectors. The success of this approach depends on the female’s behaviour, the time of exposure and the amount of PPF that can be carried by an individual. We explore the effect of PPF exposure at seven time points around blood feeding on individual Anopheles gambiae sensu stricto and Culex quinquefasciatus fecundity and ability to transfer in laboratory assays. Method Mosquitoes were exposed to 2.6 mg PPF per m2 at 48, 24 and 0.5 hours before and after a blood meal and on the day of egg-laying. The proportion of exposed females (N = 80-100) laying eggs, the number of eggs laid and hatched was studied. Transfer of PPF to oviposition cups was assessed by introducing 10 late instar insectary-reared An. gambiae s.s. larvae into all the cups and monitored for adult emergence inhibition. Results Exposure to PPF between 24 hours before and after a blood meal had significant sterilizing effects: females of both species were 6 times less likely (Odds ratio (OR) 0.16, 95% confidence interval (CI) 0.10-0.26) to lay eggs than unexposed females. Of the few eggs laid, the odds of an egg hatching was reduced 17 times (OR 0.06, 95% CI 0.04-0.08) in Anopheles but only 1.2 times (OR 0.82, 95% CI 0.73-0.93) in Culex. Adult emergence inhibition from larvae introduced in the oviposition cups was observed only from cups in which eggs were laid. When females were exposed to PPF close to egg laying they transferred enough PPF to reduce emergence by 65-71% (95% CI 62-74%). Conclusion PPF exposure within a day before and after blood feeding affects egg-development in An. gambiae s.s. and Cx. quinquefasciatus and presents a promising opportunity for integrated control of vectors and nuisance mosquitoes. However, sterilized females are

  20. Evaluating RNAlater® as a preservative for using near-infrared spectroscopy to predict Anopheles gambiae age and species

    PubMed Central

    2011-01-01

    Background Mosquito age and species identification is a crucial determinant of the efficacy of vector control programmes. Near-infrared spectroscopy (NIRS) has previously been applied successfully to rapidly, non-destructively, and simultaneously determine the age and species of freshly anesthetized African malaria vectors from the Anopheles gambiae s.l. species complex: An. gambiae s. s. and Anopheles arabiensis. However, this has only been achieved on freshly-collected specimens and future applications will require samples to be preserved between field collections and scanning by NIRS. In this study, a sample preservation method (RNAlater®) was evaluated for mosquito age and species identification by NIRS against scans of fresh samples. Methods Two strains of An. gambiae s.s. (CDC and G3) and two strains of An. arabiensis (Dongola, KGB) were reared in the laboratory while the third strain of An. arabiensis (Ifakara) was reared in a semi-field system. All mosquitoes were scanned when fresh and rescanned after preservation in RNAlater® for several weeks. Age and species identification was determined using a cross-validation. Results The mean accuracy obtained for predicting the age of young (<7 days) or old (≥ 7 days) of all fresh (n = 633) and all preserved (n = 691) mosquito samples using the cross-validation technique was 83% and 90%, respectively. For species identification, accuracies were 82% for fresh against 80% for RNAlater® preserved. For both analyses, preserving mosquitoes in RNAlater® was associated with a highly significant reduction in the likelihood of a misclassification of mosquitoes as young or old using NIRS. Important to note is that the costs for preserving mosquito specimens with RNAlater® ranges from 3-13 cents per insect depending on the size of the tube used and the number of specimens pooled in one tube. Conclusion RNAlater® can be used to preserve mosquitoes for subsequent scanning and analysis by NIRS to determine their age and

  1. Embryonic Development and Rates of Metabolic Activity in Early and Late Hatching Eggs of the Major Malaria Vector Anopheles gambiae

    PubMed Central

    Kaiser, Maria L.; Duncan, Frances D.; Brooke, Basil D.

    2014-01-01

    Anopheles gambiae eggs generally hatch at the completion of embryo development; two-three days post oviposition. However, staggered or delayed hatching has been observed whereby a single batch of eggs shows marked variation in time-to-hatch, with some eggs hatching 18 days post oviposition or later. The mechanism enabling delayed hatch has not been clearly elucidated but is likely mediated by environmental and genetic factors that either induce diapause or slow embryo development. This study aimed to compare metabolic activity and embryonic development between eggs collected from sub-colonies of the baseline Anopheles gambiae GAH colony previously selected for early or late time-to-hatch. Egg batches from early and late hatch sub-colonies as well as from the baseline colony were monitored for hatching. For both time-to-hatch selected sub-colonies and the baseline colony the majority of eggs hatched on day two post oviposition. Nevertheless, eggs produced by the late hatch sub-colony showed a significantly longer mean time to hatch than those produced by the early hatch sub-colony. The overall proportions that hatched were similar for all egg batches. CO2 output between eggs from early and late hatch sub-colonies showed significant differences only at 3 and 7 days post oviposition where eggs from the early hatch and the late hatch sub-colony were more metabolically active, respectively. No qualitative differences were observed in embryo development between the sub-colonies. It is concluded that all viable embryos develop to maturity at the same rate and that a small proportion then enter a state of diapause enabling them to hatch later. As it has previously been shown that it is possible to at least partially select for late hatch, this characteristic is likely to involve genetic as well as environmental factors. Delayed hatching in An. gambiae is likely an adaptation to maximise reproductive output despite the increased risk of desiccation in an unstable aquatic

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

    PubMed

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

    2015-12-01

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

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

    PubMed

    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. PMID:26419486

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

    PubMed Central

    2011-01-01

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

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

    PubMed Central

    2012-01-01

    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 < 0.05) in populations exposed to DDT. All mosquitoes tested were identified as 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

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

    PubMed Central

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

    2015-01-01

    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. PMID:25548172

  7. Monitoring Dry Season Persistence of Anopheles gambiae s.l. Populations in a Contained Semi-Field System in Southwestern Burkina Faso, West Africa.

    PubMed

    Mamai, W; Simard, F; Couret, D; Ouedraogo, G A; Renault, D; Dabiré, K R; Mouline, K

    2016-01-01

    To gain insight into the dry season survival strategies of Anopheles gambiae s.l., a new contained semi-field system was developed and used for the first time in Burkina Faso, West Africa. The system consisted of a screened greenhouse within which the local environment was reproduced, including all ecological requirements for mosquito development cycle completion. The system was seeded with the progenies of female Anopheles gambiae, Anopheles coluzzii, and Anopheles arabiensis collected in the vicinity of the greenhouse during the rainy season. After successful establishment in the semi-field system, mosquito populations were monitored over a 1-yr period by regular surveys of larval and adult specimens. We provided evidence for the persistence of adult mosquitoes throughout the dry season, in the absence of any suitable larval development site. During the hot and dry periods, adult insects were observed in artificial shelters (clay pots, building blocks, and dark corners). The mosquito population rapidly built up with the return of the rainy season in the area, when artificial breeding sites were refilled in the enclosure. However, only An. coluzzii and, later, An. arabiensis were detected in the subsequent rainy season, whereas no An. gambiae specimen was found. Our findings suggest that An. coluzzii and An. arabiensis may be able to aestivate throughout the dry season in Southwestern Burkina Faso, whereas An. gambiae might adopt a different dry-season survival strategy, such as long-distance re-colonization from distant locations. These results may have important implications for malaria control through targeted vector control interventions. PMID:26576935

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

    PubMed

    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

  9. Evaluation of the influence of electric nets on the behaviour of oviposition site seeking Anopheles gambiae s.s

    PubMed Central

    2014-01-01

    Background Electric nets (e-nets) are used to analyse the flight behaviour of insects and have been used extensively to study the host-oriented flight of tsetse flies. Recently we adapted this tool to analyse the oviposition behaviour of gravid malaria vectors, Anopheles gambiae s.s., orienting towards aquatic habitats and traps by surrounding an artificial pond with e-nets and collecting electrocuted mosquitoes on sticky boards on the ground next to the nets. Here we study whether e-nets themselves affect the responses of gravid An. gambiae s.s.. Methods Dual-choice experiments were carried out in 80 m2 screened semi-field systems where 200 gravid An. gambiae s.s. were released each night for 12 nights per experiment. The numbers of mosquito landing on or approaching an oviposition site were studied by adding detergent to the water in an artificial pond or surrounding the pond with a square of e-nets. We also assessed whether the supporting framework of the nets or the sticky boards used to retain electrocuted mosquitoes influenced the catch. Results Two similar detergent treated ponds presented in choice tests caught an equal proportion of the mosquitoes released, whereas a pond surrounded by e-nets caught a higher proportion than an open pond (odds ratio (OR) 1.7, 95% confidence interval (CI) 1.1 - 2.7; p < 0.017). The separate evaluation of the impact of the square of electric nets and the yellow boards on the approach of gravid females towards a pond suggests that the tower-like construction of the square of electric nets did not restrict the approach of females but the yellow sticky boards on the ground attract gravid females to a source of water (OR 2.7 95% CI 1.7 – 4.3; p < 0.001). Conclusion The trapping efficiency of the electric nets is increased when large yellow sticky boards are placed on the ground next to the e-nets to collect electrocuted mosquitoes, possibly because of increased visual contrast to the aquatic habitat. It is therefore

  10. Zoophily of Anopheles arabiensis and An. gambiae in Madagascar demonstrated by odour-baited entry traps.

    PubMed

    Duchemin, J B; Tsy, J M; Rabarison, P; Roux, J; Coluzzi, M; Costantini, C

    2001-03-01

    In Madagascar we used odour-baited entry traps (OBETs) for host choice tests of wild female anopheline mosquitoes (Diptera: Culicidae) at representative localities on the East and West sides of the island (villages Fenoarivo and Tsararano, respectively) and at the southern margin of the central plateau (Zazafotsy village, 800 m altitude). No insecticide house-spraying operations have been undertaken at these villages. Odours from a man and a calf of similar mass, concealed in different tents, were drawn by fans into separate OBETs set side by side. Traps were alternated to compensate for position effects, and different pairs of individual baits were employed for successive replicates. Totals of 266 An. funestus Giles sensu stricto and 362 An. gambiae Giles sensu lato were collected in 48 trap nights during March-June 1999. For each mosquito species the 'index of anthropophily' was defined as the proportion of females caught in the human-baited trap. For An. funestus this index was found to be consistently greater than 0.5 (value for random choice between traps/hosts), indicating that this species 'preferred' human to calf odour (index=0.83). Conversely, the index of anthropophily for An. gambiae s.l. indicated they 'chose' calf in preference to human odour (index=0.26). No significant differences of relative preference for calf or man were detected between villages; geographical variance accounted for <8% of the total experimental variance. Molecular identifications of 181 specimens of the An. gambiae complex (approximately 50% of the samples) revealed only An. arabiensis Patton at Tsararano and Zazafotsy, but >97% An. gambiae Giles sensu stricto at Fenoarivo, in accordance with prior knowledge of the differential distributions of these sibling species on the island. Predominant zoophily (i.e. intrinsic 'preference' for cattle odours) by both An. arabiensis and An. gambiae s.s. in Madagascar contrasts with their greater anthropophily in continental Africa.

  11. Field responses of Anopheles gambiae complex (Diptera: Culicidae) in Liberia using yeast-generated carbon dioxide and synthetic lure-baited light traps.

    PubMed

    Obenauer, P J; Abdel-Dayem, M S; Stoops, C A; Villinski, J T; Tageldin, R; Fahmy, N T; Diclaro, J W; Bolay, F

    2013-07-01

    Malaria infection is a serious public health problem throughout Liberia, but vector surveillance is limited or nonexistent in remote regions of the country. To better understand the spatial and temporal distribution of malaria vectors in Liberia and to support vector and malaria activities of the Liberian Ministry of Health, a study was conducted to determine the efficacy of light traps baited with a synthetic lure and CO2 for capturing Anopheles gambiae sensu lato (Giles). Traps with a ultraviolet, light-emitting diode, and incandescent lights baited with a synthetic skin lure and CO2 combinations were evaluated at four field sites in three counties of Liberia for five consecutive nights every 8 wk during 2011. In total, 4,788 mosquitoes representing 56 species from nine genera were collected throughout the 30-wk study; An. gambiae s. l. comprised 32% and of the148 An. gambiae s. s. collected, 85% were of the S form. A greater percentage of An. gambiae s. l. were collected in ultraviolet traps baited with a synthetic lure and CO2 compared with any other trap configuration. The influence of trap configuration on conclusions from surveillance efforts, specifically with regards to An. gambiae is discussed.

  12. Difluoromethyl ketones: Potent inhibitors of wild type and carbamate-insensitive G119S mutant Anopheles gambiae acetylcholinesterase.

    PubMed

    Camerino, Eugene; Wong, Dawn M; Tong, Fan; Körber, Florian; Gross, Aaron D; Islam, Rafique; Viayna, Elisabet; Mutunga, James M; Li, Jianyong; Totrov, Maxim M; Bloomquist, Jeffrey R; Carlier, Paul R

    2015-10-15

    Malaria is a devastating disease in sub-Saharan Africa, and current vector control measures are threatened by emerging resistance mechanisms. With the goal of developing new, selective, resistance-breaking insecticides we explored α-fluorinated methyl ketones as reversible covalent inhibitors of Anopheles gambiae acetylcholinesterase (AgAChE). Trifluoromethyl ketones 5 demonstrated remarkable volatility in microtiter plate assays, but 5c,e-h exhibited potent (1-100 nM) inhibition of wild type (WT) AgAChE and weak inhibition of resistant mutant G119S mutant AgAChE. Fluoromethyl ketones 10c-i exhibited submicromolar to micromolar inhibition of WT AgAChE, but again only weakly inhibited G119S AgAChE. Interestingly, difluoromethyl ketone inhibitors 9c and 9g had single digit nanomolar inhibition of WT AgAChE, and 9g had excellent potency against G119S AgAChE. Approach to steady-state inhibition was quite slow, but after 23 h incubation an IC50 value of 25.1 ± 1.2 nM was measured. We attribute the slow, tight-binding G119S AgAChE inhibition of 9g to a balance of steric size and electrophilicity. However, toxicities of 5g, 9g, and 10g to adult A. gambiae in tarsal contact, fumigation, and injection assays were lower than expected based on WT AgAChE inhibition potency and volatility. Potential toxicity-limiting factors are discussed. PMID:26386602

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

    PubMed Central

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

    2013-01-01

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

  14. Structural and Inhibitory Effects of Hinge Loop Mutagenesis in Serpin-2 from the Malaria Vector Anopheles gambiae*

    PubMed Central

    Zhang, Xin; Meekins, David A.; An, Chunju; Zolkiewski, Michal; Battaile, Kevin P.; Kanost, Michael R.; Lovell, Scott; Michel, Kristin

    2015-01-01

    Serpin-2 (SRPN2) is a key negative regulator of the melanization response in the malaria vector Anopheles gambiae. SRPN2 irreversibly inhibits clip domain serine proteinase 9 (CLIPB9), which functions in a serine proteinase cascade culminating in the activation of prophenoloxidase and melanization. Silencing of SRPN2 in A. gambiae results in spontaneous melanization and decreased life span and is therefore a promising target for vector control. The previously determined structure of SRPN2 revealed a partial insertion of the hinge region of the reactive center loop (RCL) into β sheet A. This partial hinge insertion participates in heparin-linked activation in other serpins, notably antithrombin III. SRPN2 does not contain a heparin binding site, and any possible mechanistic function of the hinge insertion was previously unknown. To investigate the function of the SRPN2 hinge insertion, we developed three SRPN2 variants in which the hinge regions are either constitutively expelled or inserted and analyzed their structure, thermostability, and inhibitory activity. We determined that constitutive hinge expulsion resulted in a 2.7-fold increase in the rate of CLIPB9Xa inhibition, which is significantly lower than previous observations of allosteric serpin activation. Furthermore, we determined that stable insertion of the hinge region did not appreciably decrease the accessibility of the RCL to CLIPB9. Together, these results indicate that the partial hinge insertion in SRPN2 does not participate in the allosteric activation observed in other serpins and instead represents a molecular trade-off between RCL accessibility and efficient formation of an inhibitory complex with the cognate proteinase. PMID:25525260

  15. Strain- and sex-specific differences in daily flight activity and the circadian clock of Anopheles gambiae mosquitoes.

    PubMed

    Rund, Samuel S C; Lee, Samuel J; Bush, Brian R; Duffield, Giles E

    2012-12-01

    Anopheles gambiae, the primary African malaria vector, is currently speciating into two incipient species, the so-called "molecular forms" M and S. While some geographic areas may contain only one form, in many areas both forms are found coexisting, but reproductively isolated. It appears that spatial segregation of mating swarms may contribute significantly to reproductive isolation as in many locales single-form swarms exist almost exclusively even though they are in close geographic proximity. The mechanism causing this spatial segregation is not well understood. Here, we compare the locomotor flight activity of M and S form male and female An. gambiae mosquitoes with the goal of identifying potential strain-specific temporal differences that could potentially serve as a mating barrier. We use an infrared beam break method to monitor flight activity of individual mosquitoes with a minute-to-minute time resolution under both LD cycle and constant dark conditions. We compare daily total flight activity, activity onset, peak in early nocturnal activity, the build up of dusk-related activity, and the free-running circadian period length. Our investigations revealed strain- and sex-specific differences in total daily activity. In both forms, males commenced nightly flight activity earlier than females, and this corresponded with a significantly shorter circadian period length in males compared to females. We note strain-specific differences in this response to dusk as males of the M form have a pronounced build up in flight activity relative to the S form males prior to complete darkness. This is likely driven by a differential response to the decreasing light intensity at dusk. We hypothesize that this behavioral difference could be a temporal factor contributing to the assembly of single-form swarms. PMID:23068991

  16. Analysis of expression in the Anopheles gambiae developing testes reveals rapidly evolving lineage-specific genes in mosquitoes

    PubMed Central

    Krzywinska, Elzbieta; Krzywinski, Jaroslaw

    2009-01-01

    Background Male mosquitoes do not feed on blood and are not involved in delivery of pathogens to humans. Consequently, they are seldom the subjects of research, which results in a very poor understanding of their biology. To gain insights into male developmental processes we sought to identify genes transcribed exclusively in the reproductive tissues of male Anopheles gambiae pupae. Results Using a cDNA subtraction strategy, five male-specifically or highly male-biased expressed genes were isolated, four of which remain unannotated in the An. gambiae genome. Spatial and temporal expression patterns suggest that each of these genes is involved in the mid-late stages of spermatogenesis. Their sequences are rapidly evolving; however, two genes possess clear homologs in a wide range of taxa and one of these probably acts in a sperm motility control mechanism conserved in many organisms, including humans. The other three genes have no match to sequences from non-mosquito taxa, thus can be regarded as orphans. RNA in situ hybridization demonstrated that one of the orphans is transcribed in spermatids, which suggests its involvement in sperm maturation. Two other orphans have unknown functions. Expression analysis of orthologs of all five genes indicated that male-biased transcription was not conserved in the majority of cases in Aedes and Culex. Conclusion Discovery of testis-expressed orphan genes in mosquitoes opens new prospects for the development of innovative control methods. The orphan encoded proteins may represent unique targets of selective anti-mosquito sterilizing agents that will not affect non-target organisms. PMID:19580678

  17. Risk Mapping of Anopheles gambiae s.l. Densities Using Remotely-Sensed Environmental and Meteorological Data in an Urban Area: Dakar, Senegal

    PubMed Central

    Machault, Vanessa; Vignolles, Cécile; Pagès, Frédéric; Gadiaga, Libasse; Tourre, Yves M.; Gaye, Abdoulaye; Sokhna, Cheikh; Trape, Jean-François; Lacaux, Jean-Pierre; Rogier, Christophe

    2012-01-01

    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. PMID:23226351

  18. 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.

    PubMed

    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.

  19. [Bio-ecological characteristics of Anopheles gambiae s.s. in irrigated rice fields of central Côte d'Ivoire].

    PubMed

    Zahouli, Z B J; Tchicaya, E S; Nsanzabana, C; Donzé, J; Utzinger, J; N'Goran, E K; Koudou, B G

    2011-12-01

    This longitudinal entomological survey was conducted between September 2008 and September 2009 in the villages of Abokro and Yaokoffikro located in an irrigated rice farming area of central Côte d'Ivoire. The purpose was to investigate the bio-ecological characteristics of Anopheles gambiae s.s. during the gonotrophic cycle. In both villages, adult mosquitoes were captured in 72 light traps, collected on humans subjects at a rate of 72 man-night from 6 p.m. to 6 a.m., and knocked down using pyrethroid spray inside 60 sentinel houses in the early morning. A total of 10,312 adult mosquitoes were collected in Abokro and 7,662 in Yaokoffikro. Anopheles was the dominant genus at both locations. Light traps were three times more efficient in Abokro than in Yaokoffikro. In both places, An. gambiae s.s. biting rates increased gradually up to a peak observed between midnight and 1 a.m. In Abokro, most An. gambiae s.s. were collected inside sleeping rooms. The endophagic rate and indoor resting density was 67.4% (n = 4798) and 14.9 females per bedroom per night, respectively, in Abokro as compared to 49.3% (n = 6775) and 2.9 females per bedroom per day, respectively, in Yaokoffikro. PMID:22393624

  20. [Bio-ecological characteristics of Anopheles gambiae s.s. in irrigated rice fields of central Côte d'Ivoire].

    PubMed

    Zahouli, Z B J; Tchicaya, E S; Nsanzabana, C; Donzé, J; Utzinger, J; N'Goran, E K; Koudou, B G

    2011-12-01

    This longitudinal entomological survey was conducted between September 2008 and September 2009 in the villages of Abokro and Yaokoffikro located in an irrigated rice farming area of central Côte d'Ivoire. The purpose was to investigate the bio-ecological characteristics of Anopheles gambiae s.s. during the gonotrophic cycle. In both villages, adult mosquitoes were captured in 72 light traps, collected on humans subjects at a rate of 72 man-night from 6 p.m. to 6 a.m., and knocked down using pyrethroid spray inside 60 sentinel houses in the early morning. A total of 10,312 adult mosquitoes were collected in Abokro and 7,662 in Yaokoffikro. Anopheles was the dominant genus at both locations. Light traps were three times more efficient in Abokro than in Yaokoffikro. In both places, An. gambiae s.s. biting rates increased gradually up to a peak observed between midnight and 1 a.m. In Abokro, most An. gambiae s.s. were collected inside sleeping rooms. The endophagic rate and indoor resting density was 67.4% (n = 4798) and 14.9 females per bedroom per night, respectively, in Abokro as compared to 49.3% (n = 6775) and 2.9 females per bedroom per day, respectively, in Yaokoffikro.

  1. Efficacy of herbal essential oils as insecticide against Aedes aegypti (Linn.), Culex quinquefasciatus (Say) and Anopheles dirus (Peyton and Harrison).

    PubMed

    Phasomkusolsil, Siriporn; Soonwera, Mayura

    2011-09-01

    The essential oils of Cananga odorata (ylang ylang), Citrus sinensis (orange), Cymbopogon citratus (lemongrass), Cymbopogon nardus (citronella grass), Eucalyptus citriodora (eucalyptus), Ocimum basilicum (sweet basil) and Syzygium aromaticum (clove), were tested for their insecticide activity against Aedes aegypti, Culex quinquefasciatus and Anopheles dirus using the WHO standard susceptibility test. These were applied in soybean oil at dose of 1%, 5% and 10% (w/v). C. citratus had the KT, values against the three mosquito species tested but the knockdown rates (at 10, 30 and 60 minutes) were lower than some essential oils. C. citratus oil had high insecticidal activity against Ae. aegypti, Cx. quinquefasciatus and An. dirus, with LC50 values of < 0.1, 2.22 and < 0.1%, respectively. Ten percent C. citratus gave the highest mortality rates (100%) 24 hours after application. This study demonstrates the potential for the essential oil of C. citratus to be used as an insecticide against 3 species of mosquitoes. PMID:22299433

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

    PubMed

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

    2016-01-01

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

  3. Ecological zones rather than molecular forms predict genetic differentiation in the malaria vector Anopheles gambiae s.s. in Ghana.

    PubMed

    Yawson, Alexander E; Weetman, David; Wilson, Michael D; Donnelly, Martin J

    2007-02-01

    The malaria mosquito Anopheles gambiae s.s. is rapidly becoming a model for studies on the evolution of reproductive isolation. Debate has centered on the taxonomic status of two forms (denoted M and S) within the nominal taxon identified by point mutations in the X-linked rDNA region. Evidence is accumulating that there are significant barriers to gene flow between these forms, but that the barriers are not complete throughout the entire range of their distribution. We sampled populations from across Ghana and southern Burkina Faso, West Africa, from areas where the molecular forms occurred in both sympatry and allopatry. Neither Bayesian clustering methods nor F(ST)-based analysis of microsatellite data found differentiation between the M and S molecular forms, but revealed strong differentiation among different ecological zones, irrespective of M/S status and with no detectable effect of geographical distance. Although no M/S hybrids were found in the samples, admixture analysis detected evidence of contemporary interform gene flow, arguably most pronounced in southern Ghana where forms occur sympatrically. Thus, in the sampled area of West Africa, lack of differentiation between M and S forms likely reflects substantial introgression, and ecological barriers appear to be of greater importance in restricting gene flow.

  4. A novel mechanism of ligand binding and release in the odorant binding protein 20 from the malaria mosquito Anopheles gambiae

    PubMed Central

    Ziemba, Brian P; Murphy, Emma J; Edlin, Hannah T; Jones, David N M

    2013-01-01

    Anopheles gambiae mosquitoes that transmit malaria are attracted to humans by the odor molecules that emanate from skin and sweat. Odorant binding proteins (OBPs) are the first component of the olfactory apparatus to interact with odorant molecules, and so present potential targets for preventing transmission of malaria by disrupting the normal olfactory responses of the insect. AgamOBP20 is one of a limited subset of OBPs that it is preferentially expressed in female mosquitoes and its expression is regulated by blood feeding and by the day/night light cycles that correlate with blood-feeding behavior. Analysis of AgamOBP20 in solution reveals that the apo-protein exhibits significant conformational heterogeneity but the binding of odorant molecules results in a significant conformational change, which is accompanied by a reduction in the conformational flexibility present in the protein. Crystal structures of the free and bound states reveal a novel pathway for entrance and exit of odorant molecules into the central-binding pocket, and that the conformational changes associated with ligand binding are a result of rigid body domain motions in α-helices 1, 4, and 5, which act as lids to the binding pocket. These structures provide new insights into the specific residues involved in the conformational adaptation to different odorants and have important implications in the selection and development of reagents targeted at disrupting normal OBP function. PMID:23081820

  5. Patterns of Mitochondrial Variation within and between African Malaria Vectors, Anopheles Gambiae and An. Arabiensis, Suggest Extensive Gene Flow

    PubMed Central

    Besansky, N. J.; Lehmann, T.; Fahey, G. T.; Fontenille, D.; Braack, LEO.; Hawley, W. A.; Collins, F. H.

    1997-01-01

    Anopheles gambiae and An. arabiensis are mosquito species responsible for most malaria transmission in sub-Saharan Africa. They are also closely related sibling species that share chromosomal and molecular polymorphisms as a consequence of incomplete lineage sorting or introgressive hybridization. To help resolve these processes, this study examined the partitioning of mtDNA sequence variation within and between species across Africa, from both population genetic and phylogeographic perspectives. Based on partial gene sequences from the cytochrome b, ND1 and ND5 genes, haplotype diversity was high but sequences were very closely related. Within species, little or no population subdivision was detected, and there was no evidence for isolation by distance. Between species, there were no fixed nucleotide differences, a high proportion of shared polymorphisms, and eight haplotypes in common over distances as great as 6000 km. Only one of 16 shared polymorphisms led to an amino acid difference, and there was no compelling evidence for nonneutral variation. Parsimony networks constructed of haplotypes from both species revealed no correspondence of haplotype with either geography or taxonomy. This trend of low intraspecific genetic divergence is consistent with evidence from allozyme and microsatellite data and is interpreted in terms of both extensive gene flow and recent range expansion from relatively large, stable populations. We argue that retention of ancestral polymorphisms is a plausible but insufficient explanation for low interspecific genetic divergence, and that extensive hybridization is a contributing factor. PMID:9409838

  6. A Comparative Genomic Analysis of Two Distant Diptera, the Fruit Fly, Drosophila melanogaster, and the Malaria Mosquito, Anopheles gambiae

    PubMed Central

    Bolshakov, Viacheslav N.; Topalis, Pantelis; Blass, Claudia; Kokoza, Elena; della Torre, Alessandra; Kafatos, Fotis C.; Louis, Christos

    2002-01-01

    Genome evolution entails changes in the DNA sequence of genes and intergenic regions, changes in gene numbers, and also changes in gene order along the chromosomes. Genes are reshuffled by chromosomal rearrangements such as deletions/insertions, inversions, translocations, and transpositions. Here we report a comparative study of genome organization in the main African malaria vector, Anopheles gambiae, relative to the recently determined sequence of the Drosophila melanogaster genome. The ancestral lines of these two dipteran insects are thought to have separated ∼250 Myr, a long period that makes this genome comparison especially interesting. Sequence comparisons have identified 113 pairs of putative orthologs of the two species. Chromosomal mapping of orthologous genes reveals that each polytene chromosome arm has a homolog in the other species. Between 41% and 73% of the known orthologous genes remain linked in the respective homologous chromosomal arms, with the remainder translocated to various nonhomologous arms. Within homologous arms, gene order is extensively reshuffled, but a limited degree of conserved local synteny (microsynteny) can be recognized. PMID:11779831

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

    PubMed Central

    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

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

    PubMed

    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.

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

    PubMed Central

    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 [AU please provide a real estimate, this seems vague]. We describe CRISPR-Cas9 endonuclease constructs that function as gene-drive systems in Anopheles gambiae, the main vector for malaria [AU:OK?]. We identified three genes (AGAP005958, AGAP011377 and AGAP007280) that confer a recessive female sterility phenotype upon disruption, and inserted into each locus CRISPR-Cas9 gene-drive constructs designed to target and edit each gene [AU:OK?]. For each locus targeted we observed strong gene drive at the molecular level, with transmission rates to progeny of 91 to 99.6%. Population modelling 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 control suppress mosquito populations to levels that do not support malaria transmission. PMID:26641531

  10. Improving the effectiveness of three essential oils against Aedes aegypti (Linn.) and Anopheles dirus (Peyton and Harrison).

    PubMed

    Auysawasdi, Nutthanun; Chuntranuluck, Sawitri; Phasomkusolsil, Siriporn; Keeratinijakal, Vichien

    2016-01-01

    Repellency of essential oil extracted from Curcuma longa, Eucalyptus globulus, and Citrus aurantium at various concentrations (5, 10, 15, 20, and 25 %) with and without 5 % vanillin was evaluated against female mosquitoes: Aedes aegypti and Anopheles dirus. The comparisons were made with a commercial chemical repellent (N,N-diethyl-3-methylbenzamide (DEET) 25 % w/w; KOR YOR 15) by arm in cage method. It was found that the essential oils with 5 % vanillin gave the longest lasting period against two mosquitoes as follows: Curcuma longa gave 150 min for Ae. aegypti, 480 min for An. dirus; Eucalyptus globulus gave 144 min for Ae. aegypti, 390 min for An. dirus; and Citrus aurantium gave 120 min for Ae. aegypti, 360 min for An. dirus. The 25 % Curcuma longa essential oil exhibited the best efficiency as equal as a commercial repellent (480 min against An. dirus). Vanillin can extend the period of time in protection against the two mosquitoes. This study indicates the potential uses of the essential oils (Curcuma longa, Eucalyptus globulus, and Citrus aurantium) with vanillin as natural mosquito repellents. PMID:26358103

  11. The effects of plant essential oils on escape response and mortality rate of Aedes aegypti and Anopheles minimus.

    PubMed

    Sathantriphop, Sunaiyana; Achee, Nicole L; Sanguanpong, Unchalee; Chareonviriyaphap, Theeraphap

    2015-12-01

    The High Throughput Screening System (HITSS) has been applied in insecticide behavioral response studies with various mosquito species. In general, chemical or natural compounds can produce a range of insect responses: contact irritancy, spatial repellency, knock-down, and toxicity. This study characterized these actions in essential oils derived from citronella, hairy basil, catnip, and vetiver in comparison to DEET and picaridin against Aedes aegypti and Anopheles minimus mosquito populations. Results indicated the two mosquito species exhibited significantly different (P<0.05) contact irritant escape responses between treatment and control for all tested compound concentrations, except with the minimum dose of picaridin (P>0.05) against Ae. aegypti. Spatial repellency responses were elicited in both mosquito species when exposed to all compounds, but the strength of the repellent response was dependent on compound and concentration. Data show that higher test concentrations had greatest toxic effects on both mosquito populations, but vetiver had no toxic effect on Ae. aegypti and picaridin did not elicit toxicity in either Ae. aegypti or An. minimus at any test concentration. Ultimately, this study demonstrates the ability of the HITSS assay to guide selection of effective plant essential oils for repelling, irritating, and killing mosquitoes. PMID:26611967

  12. Improving the effectiveness of three essential oils against Aedes aegypti (Linn.) and Anopheles dirus (Peyton and Harrison).

    PubMed

    Auysawasdi, Nutthanun; Chuntranuluck, Sawitri; Phasomkusolsil, Siriporn; Keeratinijakal, Vichien

    2016-01-01

    Repellency of essential oil extracted from Curcuma longa, Eucalyptus globulus, and Citrus aurantium at various concentrations (5, 10, 15, 20, and 25 %) with and without 5 % vanillin was evaluated against female mosquitoes: Aedes aegypti and Anopheles dirus. The comparisons were made with a commercial chemical repellent (N,N-diethyl-3-methylbenzamide (DEET) 25 % w/w; KOR YOR 15) by arm in cage method. It was found that the essential oils with 5 % vanillin gave the longest lasting period against two mosquitoes as follows: Curcuma longa gave 150 min for Ae. aegypti, 480 min for An. dirus; Eucalyptus globulus gave 144 min for Ae. aegypti, 390 min for An. dirus; and Citrus aurantium gave 120 min for Ae. aegypti, 360 min for An. dirus. The 25 % Curcuma longa essential oil exhibited the best efficiency as equal as a commercial repellent (480 min against An. dirus). Vanillin can extend the period of time in protection against the two mosquitoes. This study indicates the potential uses of the essential oils (Curcuma longa, Eucalyptus globulus, and Citrus aurantium) with vanillin as natural mosquito repellents.

  13. The effects of plant essential oils on escape response and mortality rate of Aedes aegypti and Anopheles minimus.

    PubMed

    Sathantriphop, Sunaiyana; Achee, Nicole L; Sanguanpong, Unchalee; Chareonviriyaphap, Theeraphap

    2015-12-01

    The High Throughput Screening System (HITSS) has been applied in insecticide behavioral response studies with various mosquito species. In general, chemical or natural compounds can produce a range of insect responses: contact irritancy, spatial repellency, knock-down, and toxicity. This study characterized these actions in essential oils derived from citronella, hairy basil, catnip, and vetiver in comparison to DEET and picaridin against Aedes aegypti and Anopheles minimus mosquito populations. Results indicated the two mosquito species exhibited significantly different (P<0.05) contact irritant escape responses between treatment and control for all tested compound concentrations, except with the minimum dose of picaridin (P>0.05) against Ae. aegypti. Spatial repellency responses were elicited in both mosquito species when exposed to all compounds, but the strength of the repellent response was dependent on compound and concentration. Data show that higher test concentrations had greatest toxic effects on both mosquito populations, but vetiver had no toxic effect on Ae. aegypti and picaridin did not elicit toxicity in either Ae. aegypti or An. minimus at any test concentration. Ultimately, this study demonstrates the ability of the HITSS assay to guide selection of effective plant essential oils for repelling, irritating, and killing mosquitoes.

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

    PubMed Central

    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 < 0.0001) and L1014F associated with An. arabiensis (χ2 = 11.21; P = 0.0008). The occurrence of the L1014S allele was significantly associated with lambdacyhalothrin resistance mosquitoes (Fisher exact P < 0.001). Conclusion The observed co-occurrence of L1014S and L1014F mutations coupled with reports of insecticide resistance in the country suggest that pyrethroid resistance is becoming a widespread phenomenon among our malaria vector populations. The presence of L1014F mutation in this East African mosquito population indicates the spreading of this gene across Africa. The potential operational implications of these findings on malaria control need further exploration. Objectif Les marqueurs moléculaires de la résistance aux insecticides peuvent fournir des indicateurs sensibles du développement de la résistance dans les populations de vecteurs Anopheles. Le test de ces

  15. Sugar-sensitive neurone responses and sugar feeding preferences influence lifespan and biting behaviours of the Afrotropical malaria mosquito, Anopheles gambiae.

    PubMed

    Kessler, Sébastien; Vlimant, Michèle; Guerin, Patrick M

    2015-03-01

    Floral nectar is the main source of carbohydrates for many insects including mosquitoes. Nonetheless, the physiological mechanisms underlying feeding on carbohydrates by the Afrotropical malaria mosquito Anopheles gambiae remain poorly understood. Here, we tested whether sugar sensitivity and sugar feeding preferences correlate with longevity in A. gambiae. We also tested whether feeding females on different sugar diets influences their biting behaviours. Electrophysiological recordings show that sugar neurones on the labella of females are most sensitive to sucrose, mixtures of glucose and fructose, and to melezitose; other sugars tested, including glucose and fructose presented alone, only weakly activate these taste neurones. Mosquitoes survive longest on sucrose, the most preferred sugar. Whereas feeding on a mixture of glucose and fructose is preferred over fructose or glucose alone, fructose supports higher longevity than either glucose or the mixture of the two hexoses. Females that had previously fed on glucose show a stronger biting response than those fed on sucrose, perhaps in an effort to compensate for their lower energetic state. These findings contribute to our understanding of the physiological basis of sugar feeding in A. gambiae and indicate how the sugar diet can affect laboratory-reared A. gambiae biting behaviours.

  16. Geographic population structure of the African malaria vector Anopheles gambiae suggests a role for the forest-savannah biome transition as a barrier to gene flow

    PubMed Central

    Pinto, J; Egyir-Yawson, A; Vicente, JL; Gomes, B; Santolamazza, F; Moreno, M; Charlwood, JD; Simard, F; Elissa, N; Weetman, D; Donnelly, MJ; Caccone, A; della Torre, A

    2013-01-01

    The primary Afrotropical malaria mosquito vector Anopheles gambiae sensu stricto has a complex population structure. In west Africa, this species is split into two molecular forms and displays local and regional variation in chromosomal arrangements and behaviors. To investigate patterns of macrogeographic population substructure, 25 An. gambiae samples from 12 African countries were genotyped at 13 microsatellite loci. This analysis detected the presence of additional population structuring, with the M-form being subdivided into distinct west, central, and southern African genetic clusters. These clusters are coincident with the central African rainforest belt and northern and southern savannah biomes, which suggests restrictions to gene flow associated with the transition between these biomes. By contrast, geographically patterned population substructure appears much weaker within the S-form. PMID:24062800

  17. Activity of Cinnamomum osmophloeum leaf essential oil against Anopheles gambiae s.s

    PubMed Central

    2014-01-01

    Background The increasing status of insecticide resistant mosquitoes in sub-Saharan Africa is a threatening alert to the existing control efforts. All sibling species of An. gambiae complex have evolved insecticide resistance in wild populations for different approved classes of the insecticides currently in use in the field. An alternative compound for vector control is absolutely urgently needed. In this study, the larvicidal activity and chemical composition of the Cinnamomum osmophloeum leaf essential oils were investigated. Methods C. osmophloeum leaf essential oils were extracted by hydrodistillation in a Clevenger-type apparatus for 6 hours, and their chemical compositions identified using GC-MS. These oils were evaluated against An. gambiae s.s. in both laboratory and semi-field situations. The WHO test procedures for monitoring larvicidal efficacy in malaria vectors were used. Results The composition of C. osmophloeum leaf essential oil has been found to have 11 active compounds. The most abundant compound was trans-cinnamaldehyde (70.20%) and the least abundant was caryophyllene oxide (0.08%). The larvicidal activity was found to be dosage and time dependant both in laboratory and semi-field environments with mortality ranging from 0% to 100%. The LC50 value was found to vary from 22.18 to 58.15 μg/ml in the laboratory while in semi-field environments it was 11.91 to 63.63 μg/ml. The LC90 value was found to range between 57.71 to 91.54 μg/ml in the laboratory while in semi-field environments was 52.07 to 173.77 μg/ml. Mortality ranged from 13% to 100% in the laboratory while in semi-field environments it ranged between 43% to 100% within mortality recording time intervals of 12, 24, 48, and 72 hours. Conclusions The larvicidal activity shown by C. osmophloeum leaf essential oil is a promising alternative to existing larvicides or to be incorporated in integrated larval source management compounds for An. gambiae s.s control. The efficacy

  18. Exposure to disinfectants (soap or hydrogen peroxide) increases tolerance to permethrin in Anopheles gambiae populations from the city of Yaoundé, Cameroon

    PubMed Central

    2014-01-01

    Background The rapid expansion of insecticide resistance is limiting the efficiency of malaria vector control interventions. However, current knowledge of factors inducing pyrethroid resistance remains incomplete. In the present study, the role of selection at the larval stage by disinfectants, such as soap and hydrogen peroxide (H2O2), on adult mosquito resistance to permethrin was investigated. Methods Field Anopheles gambiae sensu lato larvae, were exposed to variable concentrations of soap and H2O2. Larvae surviving to acute toxicity assays after 24 hours were reared to the adult stage and exposed to permethrin. The susceptibility level of adults was compared to the untreated control group. The effect of soap or hydrogen peroxide selection on the length of larval development and emergence rate was assessed. Result Larval bioassays analysis showed a more acute effect of hydrogen peroxide on mosquito larvae compared to soap. The regression lines describing the dose mortality profile showed higher mean and variance to hydrogen peroxide than to soap. The duration of larval development (<5 days) and adults emergence rates (1 to 77%) were shorter and lower compare to control. Anopheles gambiae s.l. larvae surviving to selection with either soap or hydrogen peroxide or both, produced adults who were up to eight-times more resistant to permethrin than mosquitoes from the untreated control group. Conclusion The present study shows that selective pressure exerted by non-insecticidal compounds such as soap and hydrogen peroxide affect An. gambiae s.l. tolerance to pyrethroids. This requires further studies with regard to the adaptation of An. gambiae s.l. to polluted habitats across sub-Saharan Africa cities. PMID:25086741

  19. Silencing of carbonic anhydrase in an Anopheles gambiae larval cell line, Ag55.

    PubMed

    Smith, Kristin E; Linser, Paul J

    2009-06-17

    RNAi has been used extensively to down-regulate proteins in adult mosquitoes; however, it is not well adapted for use in larvae. Larval mosquitoes can generate a pH as high as 10.5 in the anterior region of their midgut. The mechanisms responsible for the generation and maintenance of this pH are not entirely understood, but members of the carbonic anhydrase (CA) family of enzymes have been implicated. Here we use an An. gambiae larval cell line, Ag55 cells, to demonstrate that application of full-length double-stranded RNA specific to one CA, AgCA9, is sufficient to silence AgCA9 mRNA and down-regulate the corresponding protein. This is a first step towards determining the role(s) of these enzymes in pH regulation.

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

    PubMed

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

    2002-01-01

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

  1. Electroantennogram, flight orientation and oviposition responses of Anopheles stephensi and Aedes aegypti to a fatty acid ester-propyl octadecanoate.

    PubMed

    Seenivasagan, Thangaraj; Sharma, Kavita R; Prakash, Shri

    2012-10-01

    Studies were carried out to evaluate the role of a C(21)-fatty acid ester; propyl octadecanoate (PO) for olfaction-mediated behavioral responses of urban malaria vector, Anopheles stephensi and dengue vector, Aedes aegypti mosquitoes using electroantennogram (EAG), flight orientation and oviposition experiments. Dose dependent electrophysiological responses were recorded for PO from the antenna of both mosquito species in which 10(-5) g elicited significant EAG response. An. stephensi exhibited 2.4, 4.2 and 5.5 fold increased EAG response compared to control, while Ae. aegypti showed 1.9, 4.6 and 5.8 fold EAG responses respectively at 10(-7) g, 10(-6) g and 10(-5) g doses. In the Y-tube olfactometer, 77-80% gravid females of An. stephensi, and 64-77% of Ae. aegypti were caught in the chambers releasing 10(-6) g and 10(-5) g plume of PO. The synthetic fatty acid ester loaded onto an effervescent tablet at 0.1 mg/L, 1 mg/L and 10 mg/L elicited increased ovipositional responses from gravid mosquitoes compared to control. The oviposition activity indices (OAI) of An. stephensi females were +0.40, +0.51 and +0.58, whereas the OAI for Ae. aegypti females were +0.05, +0.36 and +0.57 respectively in 0.1, 1, 10 mg/L of PO; indicated concentration dependent increased egg deposition. Similarly, in the residual activity studies, oviposition substrates treated with PO on effervescent tablet at 1mg/L and 10mg/L received significantly increased egg deposition by gravid females of both mosquito species for up to 1 week compared to control substrates. PO can potentially be used in ovitraps to monitor An. stephensi and Ae. aegypti populations in the vector surveillance programs.

  2. Larvicidal & ovicidal efficacy of Pithecellobium dulce (Roxb.) Benth. (Fabaceae) against Anopheles stephensi Liston & Aedes aegypti Linn. (Diptera: Culicidae)

    PubMed Central

    Govindarajan, M.; Rajeswary, M.; Sivakumar, R.

    2013-01-01

    Background & objectives: In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal and ovicidal potential of the crude hexane, benzene, chloroform, ethyl acetate and methanol solvent extracts from the medicinal plant Pithecellobium dulce against the mosquito vectors, Anopheles stephensi and Aedes aegypti (Diptera: Culicidae). Methods: Larvicidal activity of P. dulce plant extracts was studied in the range of 60 to 450 mg/l against early third instar larvae of An. stephensi and Ae. aegypti in the laboratory. The larval mortality was observed after 24 h of exposure. The ovicidal activity was determined against An. stephensi and Ae. aegypti to various concentrations ranging from 100 to 750 mg/l under the laboratory conditions. Mean per cent hatchability of the eggs were observed after 48 h post treatment. Results: All leaf and seed extracts showed moderate larvicidal and ovicidal effects; however, the highest larval mortality was found in methanol extract of leaf of P. dulce against the larvae of An. stephensi and Ae. aegypti with the LC50 and LC90 values 145.43, 155.78 mg/l and 251.23, 279.73 mg/l, respectively. The per cent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. Zero hatchability was observed at 400 mg/l for leaf methanol extract and 625 mg/l for seed methanol extract of P. dulce against An. stephensi and Ae. aegypti, respectively. Compared to leaf extracts, seed extracts have low potency against the two mosquitoes. Interpretation & conclusions: The present results suggest that the leaf and seed extracts of P. dulce have the potential to be used as an ideal eco-friendly approach for the control of mosquitoes. PMID:24056567

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

    PubMed Central

    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

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

    PubMed

    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

  5. Differential effects of inbreeding and selection on male reproductive phenotype associated with the colonization and laboratory maintenance of Anopheles gambiae

    PubMed Central

    2014-01-01

    Background Effective mating between laboratory-reared males and wild females is paramount to the success of vector control strategies aiming to decrease disease transmission via the release of sterile or genetically modified male mosquitoes. However mosquito colonization and laboratory maintenance have the potential to negatively affect male genotypic and phenotypic quality through inbreeding and selection, which in turn can decrease male mating competitiveness in the field. To date, very little is known about the impact of those evolutionary forces on the reproductive biology of mosquito colonies and how they ultimately affect male reproductive fitness. Methods Here several male reproductive physiological traits likely to be affected by inbreeding and selection following colonization and laboratory rearing were examined. Sperm length, and accessory gland and testes size were compared in male progeny from field-collected females and laboratory strains of Anopheles gambiae sensu stricto colonized from one to over 25 years ago. These traits were also compared in the parental and sequentially derived, genetically modified strains produced using a two-phase genetic transformation system. Finally, genetic crosses were performed between strains in order to distinguish the effects of inbreeding and selection on reproductive traits. Results Sperm length was found to steadily decrease with the age of mosquito colonies but was recovered in refreshed strains and crosses between inbred strains therefore incriminating inbreeding costs. In contrast, testes size progressively increased with colony age, whilst accessory gland size quickly decreased in males from colonies of all ages. The lack of heterosis in response to crossing and strain refreshing in the latter two reproductive traits suggests selection for insectary conditions. Conclusions These results show that inbreeding and selection differentially affect reproductive traits in laboratory strains overtime and that

  6. Changes in transcript abundance for cuticular proteins and other genes three hours after a blood meal in Anopheles gambiae

    PubMed Central

    Vannini, Laura; Dunn, W. Augustine; Reed, Tyler W.; Willis, Judith H.

    2014-01-01

    Numerous studies have examined changes in transcript levels after Anopheles gambiae takes a blood meal. Marinotti et al. (2006) used microarrays and reported massive changes in transcript levels 3 h after feeding (BF3h) compared to non-blood fed (NBF). We were intrigued by the number of transcripts for structural cuticular proteins (CPs) that showed such major differences in levels and employed paired-end (50 bp) RNA-seq technology to compare whole body transcriptomes from 5-day-old females NBF and BF3h. We detected transcripts for the majority of CPs (164/243) but levels of only 12 were significantly altered by the blood meal. While relative transcript levels of NBF females were somewhat similar to the microarray data, there were major differences in BF3h animals, resulting in levels of many transcripts, both for CPs and other genes changing in the opposite direction. We compared our data also to other studies done with both microarrays and RNA-seq. Findings were consistent that a small number of CP genes have transcripts that persist even in 5-day-old adults. Some of these transcripts showed diurnal rhythms (Rund et al., 2013; Rinker et al., 2013). In situ hybridization revealed that transcripts for several of these CP genes were found exclusively or predominantly in the eye. Transcripts other than for CPs that changed in response to blood-feeding were predominantly expressed in midgut and Malpighian tubules. Even in these tissues, genes responsible for proteins with similar functions, such as immunity or digestion, responded differently, with transcript levels for some rising and others falling. These data demonstrate that genes coding for some CPs are dynamic in expression even in adults and that the response to a blood meal is rapid and precisely orchestrated. PMID:24269292

  7. Changes in transcript abundance for cuticular proteins and other genes three hours after a blood meal in Anopheles gambiae.

    PubMed

    Vannini, Laura; Augustine Dunn, W; Reed, Tyler W; Willis, Judith H

    2014-01-01

    Numerous studies have examined changes in transcript levels after Anopheles gambiae takes a blood meal. Marinotti et al. (2006) used microarrays and reported massive changes in transcript levels 3 h after feeding (BF3h) compared to non-blood fed (NBF). We were intrigued by the number of transcripts for structural cuticular proteins (CPs) that showed such major differences in levels and employed paired-end (50 bp) RNA-seq technology to compare whole body transcriptomes from 5-day-old females NBF and BF3h. We detected transcripts for the majority of CPs (164/243) but levels of only 12 were significantly altered by the blood meal. While relative transcript levels of NBF females were somewhat similar to the microarray data, there were major differences in BF3h animals, resulting in levels of many transcripts, both for CPs and other genes changing in the opposite direction. We compared our data also to other studies done with both microarrays and RNA-seq. Findings were consistent that a small number of CP genes have transcripts that persist even in 5-day-old adults. Some of these transcripts showed diurnal rhythms (Rund et al., 2013; Rinker et al., 2013). In situ hybridization revealed that transcripts for several of these CP genes were found exclusively or predominantly in the eye. Transcripts other than for CPs that changed in response to blood-feeding were predominantly expressed in midgut and Malpighian tubules. Even in these tissues, genes responsible for proteins with similar functions, such as immunity or digestion, responded differently, with transcript levels for some rising and others falling. These data demonstrate that genes coding for some CPs are dynamic in expression even in adults and that the response to a blood meal is rapid and precisely orchestrated.

  8. Expression and characterization of an epoxide hydrolase from Anopheles gambiae with high activity on epoxy fatty acids

    PubMed Central

    Xu, Jiawen; Morisseau, Christophe; Hammock, Bruce D.

    2014-01-01

    In insects, epoxide hydrolases (EHs) play critical roles in the metabolism of xenobiotic epoxides from the food resources and in the regulation of endogenous chemical mediators, such as juvenile hormones. Using the baculovirus expression system, we expressed and characterized an epoxide hydrolase from Anopheles gambiae (AgEH) that is distinct in evolutionary history from insect juvenile hormone epoxide hydrolases (JHEHs). We partially purified the enzyme by ion exchange chromatography and isoelectric focusing. The experimentally determined molecular weight and pI were estimated to be 35kD and 6.3 respectively, different than the theoretical ones. The AgEH had the greatest activity on long chain epoxy fatty acids such as 14,15-epoxyeicosatrienoic acids (14,15-EET) and 9,10-epoxy-12Z-octadecenoic acids (9,10-EpOME or leukotoxin) among the substrates evaluated. Juvenile hormone III, a terpenoid insect growth regulator, was the next best substrate tested. The AgEH showed kinetics comparable to the mammalian soluble epoxide hydrolases, and the activity could be inhibited by AUDA [12-(3-adamantan-1-yl-ureido) dodecanoic acid], a urea-based inhibitor designed to inhibit the mammalian soluble epoxide hydrolases. The rabbit serum generated against the soluble epoxide hydrolase of Mus musculus can both cross-react with natural and denatured forms of the AgEH, suggesting immunologically they are similar. The study suggests there are mammalian sEH homologs in insects, and epoxy fatty acids may be important chemical mediators in insects. PMID:25173592

  9. MalariaSphere: A greenhouse-enclosed simulation of a natural Anopheles gambiae (Diptera: Culicidae) ecosystem in western Kenya

    PubMed Central

    Knols, Bart GJ; Njiru, Basilio N; Mathenge, Evan M; Mukabana, Wolfgang R; Beier, John C; Killeen, Gerry F

    2002-01-01

    Background The development and implementation of innovative vector control strategies for malaria control in Africa requires in-depth ecological studies in contained semi-field environments. This particularly applies to the development and release of genetically-engineered vectors that are refractory to Plasmodium infection. Here we describe a modified greenhouse, designed to simulate a natural Anopheles gambiae Giles ecosystem, and the first successful trials to complete the life-cycle of this mosquito vector therein. Methods We constructed a local house, planted crops and created breeding sites to simulate the natural ecosystem of this vector in a screen-walled greenhouse, exposed to ambient climate conditions, in western Kenya. Using three different starting points for release (blood-fed females, virgin females and males, or eggs), we allowed subsequent stages of the life-cycle to proceed under close observation until one cycle was completed. Results Completion of the life-cycle was observed in all three trials, indicating that the major life-history behaviours (mating, sugar feeding, oviposition and host seeking) occurred successfully. Conclusion The system described can be used to study the behavioural ecology of laboratory-reared and wild mosquitoes, and lends itself to contained studies on the stability of transgenes, fitness effects and phenotypic characteristics of genetically-engineered disease vectors. The extension of this approach, to enable continuous maintenance of successive and overlapping insect generations, should be prioritised. Semi-field systems represent a promising means to significantly enhance our understanding of the behavioural and evolutionary ecology of African malaria vectors and our ability to develop and evaluate innovative control strategies. With regard to genetically-modified mosquitoes, development of such systems is an essential prerequisite to full field releases. PMID:12537599

  10. Semi-Field Evaluation of Metofluthrin-Impregnated Nets on Host-Seeking Aedes aegypti and Anopheles dirus.

    PubMed

    Ponlawat, Alongkot; Kankaew, Prasan; Chanaimongkol, Somporn; Pongsiri, Arissara; Richardson, Jason H; Evans, Brian P

    2016-06-01

    The efficacy of a metofluthrin-impregnated net (MIN) known as the "Mushikonazu" on the house entry behavior of female Aedes aegypti and Anopheles dirus mosquitoes was evaluated using a semi-field 50-m tunnel setup. While the MIN is labeled for the control of chironomids and moth flies, this study determined the feasibility of using the device, given its current construction and metofluthrin formulation, as a spatial repellent against mosquitoes. Sentinel and cone bioassays were used to determine the insecticidal effect of the MIN. A spatial activity index (SAI) was calculated to evaluate responses of the mosquitoes. For the spatial repellent evaluation against Ae. aegypti, the overall mean of SAI was slightly less than 0 at wk 1 after the MIN application and then decreased for the last 4 wk showing a preference to treatment tent. For An. dirus, the mean SAI at wk 1 was positive, indicating a presumed repellent effect of the MIN against An. dirus. For the subsequent 4 wk, the SAI was negative, indicating a preference for the MIN. Results suggested that the MIN may not be a promising approach to repel Ae. aegypti and An. dirus under field conditions in Thailand. However, it remains probable that the MIN may be effective as a spatial repellent if modifications are made to the metofluthrin concentration or formulation and/or the construction of the device.

  11. Semi-Field Evaluation of Metofluthrin-Impregnated Nets on Host-Seeking Aedes aegypti and Anopheles dirus.

    PubMed

    Ponlawat, Alongkot; Kankaew, Prasan; Chanaimongkol, Somporn; Pongsiri, Arissara; Richardson, Jason H; Evans, Brian P

    2016-06-01

    The efficacy of a metofluthrin-impregnated net (MIN) known as the "Mushikonazu" on the house entry behavior of female Aedes aegypti and Anopheles dirus mosquitoes was evaluated using a semi-field 50-m tunnel setup. While the MIN is labeled for the control of chironomids and moth flies, this study determined the feasibility of using the device, given its current construction and metofluthrin formulation, as a spatial repellent against mosquitoes. Sentinel and cone bioassays were used to determine the insecticidal effect of the MIN. A spatial activity index (SAI) was calculated to evaluate responses of the mosquitoes. For the spatial repellent evaluation against Ae. aegypti, the overall mean of SAI was slightly less than 0 at wk 1 after the MIN application and then decreased for the last 4 wk showing a preference to treatment tent. For An. dirus, the mean SAI at wk 1 was positive, indicating a presumed repellent effect of the MIN against An. dirus. For the subsequent 4 wk, the SAI was negative, indicating a preference for the MIN. Results suggested that the MIN may not be a promising approach to repel Ae. aegypti and An. dirus under field conditions in Thailand. However, it remains probable that the MIN may be effective as a spatial repellent if modifications are made to the metofluthrin concentration or formulation and/or the construction of the device. PMID:27280351

  12. Microarray and RNAi Analysis of P450s in Anopheles gambiae Male and Female Steroidogenic Tissues: CYP307A1 Is Required for Ecdysteroid Synthesis

    PubMed Central

    Pondeville, Emilie; David, Jean-Philippe; Guittard, Emilie; Maria, Annick; Ranson, Hilary

    2013-01-01

    In insects, the steroid hormone 20-hydroxyecdysone (20E) coordinates major developmental transitions. While the first and the final steps of 20E biosynthesis are characterized, the pathway from 7-dehydrocholesterol to 5β-ketodiol, commonly referred as the “black box”, remains hypothetical and whether there are still unidentified enzymes is unknown. The black box would include some oxidative steps, which are believed to be mediated by P450 enzymes. To identify new enzyme(s) involved in steroid synthesis, we analyzed by small-scale microarray the expression of all the genes encoding P450 enzymes of the malaria mosquito Anopheles gambiae in active steroidogenic organs of adults, ovaries from blood-fed females and male reproductive tracts, compared to inactive steroidogenic organs, ovaries from non-blood-fed females. Some genes encoding P450 enzymes were specifically overexpressed in female ovaries after a blood-meal or in male reproductive tracts but only three genes were found to be overexpressed in active steroidogenic organs of both females and males: cyp307a1, cyp4g16 and cyp6n1. Among these genes, only cyp307a1 has an expression pattern similar to other mosquito steroidogenic genes. Moreover, loss-of-function by transient RNAi targeting cyp307a1 disrupted ecdysteroid production demonstrating that this gene is required for ecdysteroid biosynthesis in Anopheles gambiae. PMID:24324583

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

    PubMed

    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. PMID:25630696

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

    PubMed

    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.

  15. Short report: A new polymerase chain reaction-restriction fragment length polymorphism method to identify Anopheles arabiensis from An. gambiae and its two molecular forms from degraded DNA templates or museum samples.

    PubMed

    Santolamazza, Federica; Della Torre, Alessandra; Caccone, Adalgisa

    2004-06-01

    We present a polymerase chain reaction-restriction fragment length polymorphism method to simultaneously distinguish the two Anopheles gambiae M and S molecular forms and Anopheles arabiensis. This method uses different diagnostic sites than previously published methods, and it is based on the amplification of a smaller ribosomal DNA fragment. We have tested this protocol in a variety of samples from different geographic regions and various ages of preservation to ascertain the robustness of this protocol over a wide geographic window and on DNA templates of poor quality. This procedure is as efficient as previous ones in discriminating An. arabiensis from the two taxa in An gambiae s.s. However, it performs better than others on poor quality templates such as the ones from museum collections, and poorly stored field collected material. However, it must be noted that it does not allow the simultaneous discrimination of all the species in the An. gambiae complex.

  16. Larvicidal activity of Morinda citrifolia L. (Noni) (Family: Rubiaceae) leaf extract against Anopheles stephensi, Culex quinquefasciatus, and Aedes aegypti.

    PubMed

    Kovendan, Kalimuthu; Murugan, Kadarkarai; Shanthakumar, Shanmugam Perumal; Vincent, Savariar; Hwang, Jiang-Shiou

    2012-10-01

    Morinda citrifolia leaf extract was tested for larvicidal activity against three medically important mosquito vectors such as malarial vector Anopheles stephensi, dengue vector Aedes aegypti, and filarial vector Culex quinquefasciatus. The plant material was shade dried at room temperature and powdered coarsely. From the leaf, 1-kg powder was macerated with 3.0 L of hexane, chloroform, acetone, methanol, and water sequentially for a period of 72 h each and filtered. The yield of extracts was hexane (13.56 g), chloroform (15.21 g), acetone (12.85 g), methanol (14.76 g), and water (12.92 g), respectively. The extracts were concentrated at reduced temperature on a rotary vacuum evaporator and stored at a temperature of 4°C. The M. citrifolia leaf extract at 200, 300, 400, 500, and 600 ppm caused a significant mortality of three mosquito species. Hexane, chloroform, acetone, and water caused moderate considerable mortality; however, the highest larval mortality was methanolic extract, observed in three mosquito vectors. The larval mortality was observed after 24-h exposure. No mortality was observed in the control. The third larvae of Anopheles stephensi had values of LC(50) = 345.10, 324.26, 299.97, 261.96, and 284.59 ppm and LC(90) = 653.00, 626.58, 571.89, 505.06, and 549.51 ppm, respectively. The Aedes aegypti had values of LC(50) = 361.75, 343.22, 315.40, 277.92, and 306.98 ppm and LC(90) = 687.39, 659.02, 611.35, 568.18, and 613.25 ppm, respectively. The Culex quinquefasciatus had values of LC(50) = 382.96, 369.85, 344.34, 330.42, and 324.64 ppm and LC(90) = 726.18, 706.57, 669.28, 619.63, and 644.47 ppm, respectively. The results of the leaf extract of M. citrifolia are promising as good larvicidal activity against the mosquito vector Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. This is a new eco-friendly approach for the control of vector control programs. Therefore, this study provides first report on the larvicidal activities against three

  17. How reliable are light traps in estimating biting rates of adult Anopheles gambiae s.l. (Diptera: Culicidae) in the presence of treated bed nets?

    PubMed

    Magbity, E B; Magbity, E B; Lines, J D; Marbiah, M T; David, K; Peterson, E

    2002-02-01

    The sampling efficiency of light trap catches relative to human bait catches in estimating biting rates of the mosquito Anopheles gambiae Giles was investigated in two types of community in southern Sierra Leone: (i) where most of the inhabitants slept under treated bed nets; and (ii) where most of the inhabitants slept without bed nets. The number of female A. gambiae mosquitoes caught in these communities by light trap was strongly correlated (r > or = 0.72) with those from corresponding human biting catches performed either on the same or adjacent nights. It was found that the relative sampling efficiency of light traps varied slightly but significantly with mosquito abundance in villages with treated bed nets, but not in those without them. Nevertheless, the relationship between relative sampling efficiency and mosquito abundance did not differ significantly between the two types of village. Overall, there was insufficient evidence to show that the presence of treated nets altered the relative efficiency of light traps and any bias was only slight, and unlikely to be of any practical importance. Hence, it was concluded that light traps can be used as a surrogate for human bait catches in estimating biting rates of A. gambiae mosquitoes in the two communities.

  18. 3-Oxoisoxazole-2(3H)-carboxamides and isoxazol-3-yl carbamates: Resistance-breaking acetylcholinesterase inhibitors targeting the malaria mosquito, Anopheles gambiae

    PubMed Central

    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

  19. Mapping the ranges and relative abundance of the two principal African malaria vectors, Anopheles gambiae sensu stricto and An. arabiensis, using climate data.

    PubMed Central

    Lindsay, S W; Parson, L; Thomas, C J

    1998-01-01

    Members of the Anopheles gambiae complex are major malaria vectors in Africa. We tested the hypothesis that the range and relative abundance of the two major vectors in the complex, An. gambiae sensu stricto and An. arabiensis, could be defined by climate. Climate was characterized at mosquito survey sites by extracting data for each location from climate surfaces using a Geographical Information System. Annual precipitation, together with annual and wet season temperature, defined the ranges of both vectors and were used to map suitable climate zones. Using data from West Africa, we found that where the species were sympatric, An. gambiae s.s. predominated in saturated environments, and An. arabiensis was more common in sites subject to desiccation (r2 = 0.875, p < 0.001). We used the nonlinear equation that best described this relationship to map habitat suitability across Africa. This simple model predicted accurately the relative abundance of both vectors in Tanzania (rs = 0.745, p = 0.002), where species composition is highly variable. The combined maps of species' range and relative abundance showed very good agreement with published maps. This technique represents a new approach to mapping the distribution of malaria vectors over large areas and may facilitate species-specific vector control activities. PMID:9633110

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

    USGS Publications Warehouse

    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 < 0.01). Survival was greater when sugar sources were randomly

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

    PubMed

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

    2013-04-01

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

  2. Biophysical Analysis of Anopheles gambiae Leucine-Rich Repeat Proteins APL1A1, APL1B and APL1C and Their Interaction with LRIM1

    SciTech Connect

    Williams, Marni; Summers, Brady J.; Baxter, Richard H. G.; Kobe, Bostjan

    2015-03-16

    Natural infection of Anopheles gambiae by malaria-causing Plasmodium parasites is significantly influenced by the APL1 genetic locus. The locus contains three closely related leucine-rich repeat (LRR) genes, APL1A, APL1B and APL1C. Multiple studies have reported the participation of APL1A—C in the immune response of A. gambiae to invasion by both rodent and human Plasmodium isolates. APL1C forms a heterodimer with the related LRR protein LRIM1 via a C-terminal coiled-coil domain that is also present in APL1A and APL1B. The LRIM1/APL1C heterodimer protects A. gambiae from infection by binding the complement-like protein TEP1 to form a stable and active immune complex. We report solution x-ray scatting data for the LRIM1/APL1C heterodimer, the oligomeric state of LRIM1/APL1 LRR domains in solution and the crystal structure of the APL1B LRR domain. The LRIM1/APL1C heterodimeric complex has a flexible and extended structure in solution. In contrast to the APL1A, APL1C and LRIM1 LRR domains, the APL1B LRR domain is a homodimer. The crystal structure of APL1B-LRR shows that the homodimer is formed by an N-terminal helix that complements for the absence of an N-terminal capping motif in APL1B, which is a unique distinction within the LRIM1/APL1 protein family. Full-length APL1A1 and APL1B form a stable complex with LRIM1. Our results support a model in which APL1A1, APL1B and APL1C can all form an extended, flexible heterodimer with LRIM1, providing a repertoire of functional innate immune complexes to protect A. gambiae from a diverse array of pathogens.

  3. Biophysical Analysis of Anopheles gambiae Leucine-Rich Repeat Proteins APL1A1, APL1B and APL1C and Their Interaction with LRIM1

    DOE PAGES

    Williams, Marni; Summers, Brady J.; Baxter, Richard H. G.; Kobe, Bostjan

    2015-03-16

    Natural infection of Anopheles gambiae by malaria-causing Plasmodium parasites is significantly influenced by the APL1 genetic locus. The locus contains three closely related leucine-rich repeat (LRR) genes, APL1A, APL1B and APL1C. Multiple studies have reported the participation of APL1A—C in the immune response of A. gambiae to invasion by both rodent and human Plasmodium isolates. APL1C forms a heterodimer with the related LRR protein LRIM1 via a C-terminal coiled-coil domain that is also present in APL1A and APL1B. The LRIM1/APL1C heterodimer protects A. gambiae from infection by binding the complement-like protein TEP1 to form a stable and active immune complex.more » We report solution x-ray scatting data for the LRIM1/APL1C heterodimer, the oligomeric state of LRIM1/APL1 LRR domains in solution and the crystal structure of the APL1B LRR domain. The LRIM1/APL1C heterodimeric complex has a flexible and extended structure in solution. In contrast to the APL1A, APL1C and LRIM1 LRR domains, the APL1B LRR domain is a homodimer. The crystal structure of APL1B-LRR shows that the homodimer is formed by an N-terminal helix that complements for the absence of an N-terminal capping motif in APL1B, which is a unique distinction within the LRIM1/APL1 protein family. Full-length APL1A1 and APL1B form a stable complex with LRIM1. Our results support a model in which APL1A1, APL1B and APL1C can all form an extended, flexible heterodimer with LRIM1, providing a repertoire of functional innate immune complexes to protect A. gambiae from a diverse array of pathogens.« less

  4. Biophysical Analysis of Anopheles gambiae Leucine-Rich Repeat Proteins APL1A1, APL1B and APL1C and Their Interaction with LRIM1

    PubMed Central

    Williams, Marni; Summers, Brady J.; Baxter, Richard H. G.

    2015-01-01

    Natural infection of Anopheles gambiae by malaria-causing Plasmodium parasites is significantly influenced by the APL1 genetic locus. The locus contains three closely related leucine-rich repeat (LRR) genes, APL1A, APL1B and APL1C. Multiple studies have reported the participation of APL1A—C in the immune response of A. gambiae to invasion by both rodent and human Plasmodium isolates. APL1C forms a heterodimer with the related LRR protein LRIM1 via a C-terminal coiled-coil domain that is also present in APL1A and APL1B. The LRIM1/APL1C heterodimer protects A. gambiae from infection by binding the complement-like protein TEP1 to form a stable and active immune complex. Here we report solution x-ray scatting data for the LRIM1/APL1C heterodimer, the oligomeric state of LRIM1/APL1 LRR domains in solution and the crystal structure of the APL1B LRR domain. The LRIM1/APL1C heterodimeric complex has a flexible and extended structure in solution. In contrast to the APL1A, APL1C and LRIM1 LRR domains, the APL1B LRR domain is a homodimer. The crystal structure of APL1B-LRR shows that the homodimer is formed by an N-terminal helix that complements for the absence of an N-terminal capping motif in APL1B, which is a unique distinction within the LRIM1/APL1 protein family. Full-length APL1A1 and APL1B form a stable complex with LRIM1. These results support a model in which APL1A1, APL1B and APL1C can all form an extended, flexible heterodimer with LRIM1, providing a repertoire of functional innate immune complexes to protect A. gambiae from a diverse array of pathogens. PMID:25775123

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

    PubMed

    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

  6. The Anopheles gambiae cE5 salivary protein: a sensitive biomarker to evaluate the efficacy of insecticide-treated nets in malaria vector control.

    PubMed

    Marie, Alexandra; Ronca, Raffaele; Poinsignon, Anne; Lombardo, Fabrizio; Drame, Papa M; Cornelie, Sylvie; Besnard, Patrick; Le Mire, Jacques; Fiorentino, Gabriella; Fortes, Filomeno; Carnevale, Pierre; Remoue, Franck; Arcà, Bruno

    2015-06-01

    Evaluation of vector control is crucial for improving malaria containment and, according to World Health Organization, new complementary indicators would be very valuable. In this study the IgG response to the Anopheles-specific cE5 salivary protein was tested as a tool to evaluate the efficacy of insecticide-treated nets in reducing human exposure to malaria vectors. Sera collected during a longitudinal study carried out in Angola, and including entomological and parasitological data, were used to assess the IgG response to the Anopheles gambiae cE5 in both children and adults, before and after the application of insecticide-treated nets. Seasonal fluctuation of specific IgG antibody levels according to exposure was only found in children (up to ≈ 14 years old) whose anti-cE5 IgG response dropped after bed nets installation. These results were fully consistent with previous findings obtained with the same set of sera and indicating a substantial reduction of human-vector contact shortly after nets implementation. Overall, children IgG response to the cE5 protein appeared a very sensitive biomarker, which allowed for the detection of even weak exposure to Anopheles bites, indicating it may represent a reliable additional tool to evaluate the efficacy of vector control interventions.

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

    PubMed Central

    2011-01-01

    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 Progress in understanding

  8. Effect of sugar on male Anopheles gambiae mating performance, as modified by temperature, space, and body size

    PubMed Central

    Gary, Richard E; Cannon, James W; Foster, Woodbridge A

    2009-01-01

    Background Anopheles gambiae plant-sugar feeding was thought to be rare and physiologically optional. Unlike adult females, males have no alternative source of energy and soon die with only water, yet they might be competent to inseminate all females within their brief lifespan. This study was designed to detect sugar's effect, if any, on male performance. Methods Males with and without 20% sucrose were evaluated at two body sizes and two temperatures, 23° and 27°C. Survival was recorded twice daily, and sexual behaviour was recorded each night after adult emergence. Insemination at a 2:1 male:female ratio was examined in three cage sizes, including walk-in mesocosms. Results Without sugar, males of both sizes lived longer at 23° than 27°C, and large males lived longer at each temperature. Survival of large males at low temperature averaged 3.7 days, small males at high temperature, 1.9 days. With sugar, males in all four treatments suffered minimal mortality. With sugar, in small cages, large males at 27°C matured most rapidly. A few erected fibrillae and inseminated females on night 1. On night 2, maximal proportions erected fibrillae and swarmed, and over one-third of females became inseminated. Small sugar-fed males at 23°C matured most slowly but had achieved nearly maximal levels of swarming by night 3. By night 5, small males had inseminated more than half the females, and large males had inseminated nearly all of them. Without sugar, large males progressed similarly during the first two nights. On night 3, however, the proportion erecting fibrillae and swarming declined precipitously at 27°C, and to a lesser degree at 23°C. Cumulative insemination never reached high levels. Small males never achieved high levels of fibrillar erection or swarming and inseminated few females, even at 23°C. In larger cages and under more semi-natural conditions, regardless of body size, without sugar male insemination capacity was virtually nonexistent. Conclusion

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

    PubMed Central

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

    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, 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 4th 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 2nd and 3rd instars of Ae. aegypti and 145.48±11.76, 188.10±12.92 against 2nd and 3rd instars of An. stephensi, respectively. Crude protein from the water extract was precipitated using acetone and tested against 2nd, 3rd and 4th 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 2nd, 3rd and 4th instars of Ae. aegypti and An. stephensi, respectively. Conclusion: 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. PMID:25629069

  10. Behavioral responses of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus against various synthetic and natural repellent compounds.

    PubMed

    Sathantriphop, Sunaiyana; White, Sabrina A; Achee, Nicole L; Sanguanpong, Unchalee; Chareonviriyaphap, Theeraphap

    2014-12-01

    The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito-repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65-98% escape for contact, 21.4-94.4% escape for non-contact) compared to Ae. aegypti (3.7-72.2% escape (contact), 0-31.7% (non-contact)) and Ae. albopictus (3.5-94.4% escape (contact), 11.2-63.7% (non-contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65-97.8% escape) than non-contact repellents (0-50.8% escape for non-contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes.

  11. Behavioral responses of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus against various synthetic and natural repellent compounds.

    PubMed

    Sathantriphop, Sunaiyana; White, Sabrina A; Achee, Nicole L; Sanguanpong, Unchalee; Chareonviriyaphap, Theeraphap

    2014-12-01

    The behavioral responses of colony populations of Aedes aegypti, Aedes albopictus, Culex quinquefasciatus, and Anopheles minimus to four essential oils (citronella, hairy basil, catnip, and vetiver), two standard repellents (DEET and picaridin), and two synthetic pyrethroids (deltamethrin and permethrin) were conducted in the laboratory using an excito-repellency test system. Results revealed that Cx. quinquefasciatus and An. minimus exhibited much stronger behavioral responses to all test compounds (65-98% escape for contact, 21.4-94.4% escape for non-contact) compared to Ae. aegypti (3.7-72.2% escape (contact), 0-31.7% (non-contact)) and Ae. albopictus (3.5-94.4% escape (contact), 11.2-63.7% (non-contact)). In brief, essential oil from vetiver elicited the greatest irritant responses in Cx. quinquefasciatus (96.6%) and An. minimus (96.5%) compared to the other compounds tested. The synthetic pyrethroids caused a stronger contact irritant response (65-97.8% escape) than non-contact repellents (0-50.8% escape for non-contact) across all four mosquito species. Picaridin had the least effect on all mosquito species. Findings from the current study continue to support the screening of essential oils from various plant sources for protective properties against field mosquitoes. PMID:25424262

  12. Adulticidal and smoke toxicity of Cipadessa baccifera (Roth) plant extracts against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus.

    PubMed

    Ramkumar, Govindaraju; Karthi, Sengodan; Muthusamy, Ranganathan; Natarajan, Devarajan; Shivakumar, Muthugounder S

    2015-01-01

    Mosquito vectors are responsible for the transmission of parasitic and viral infections, including loss in commercial and labor outputs, particularly in developing countries with tropical and subtropical climates. The aim of the present study is to evaluate the adulticidal and smoke toxicity of Cipadessa baccifera (Roth) against three important mosquitoes vectors, Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae). 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 acetone extract against An. stephensi followed by Ae. aegypti and Cx. quinquefasciatus with the LD50 and LD90 values 16.021 (14.080-18.345), 29.095 (25.118-34.089); 23.581 (22.100-28.315), 38.636 (35.321-41.021); and 13.560 (9.479-17.391), 248.35 (203.47-344.43) mg/ml, respectively. No mortality was recorded in the control. Smoke toxicity was observed at 10-min interval for 40 min and the mortality data were recorded. Among the C. baccifera plant powder tested. Smoke toxicity results show that Cx. quinquefasciatus, An. stephensi, and Ae. aegypti shows 88.6 ± 1.8, 78.2 ± 0.5, and 77 ± 1, respectively. One hundred percent mortality was recorded in the commercial mosquito control. The present study shows that C. baccifera leaf powder can be used as an efficient toxicity against mosquitoes. These results suggest that the leaf extracts of C. baccifera have a potential to be used as an ideal eco-friendly approach for the control of mosquitoes. PMID:25320044

  13. Prodigiosin produced by Serratia marcescens NMCC46 as a mosquito larvicidal agent against Aedes aegypti and Anopheles stephensi.

    PubMed

    Patil, Chandrashekhar D; Patil, Satish V; Salunke, Bipinchandra K; Salunkhe, Rahul B

    2011-10-01

    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 potential of microbial pigment prodigiosin produced by Serratia marcescens NMCC46 against Aedes aegypti and Anopheles stephensi. The pigment of S. marcescens NMCC46 was extracted after 24 h from mannitol containing nutrient broth media. The effects of crude extracted pigment on the growth, survival, development, and other life cycle aspects were studied. The LC(50) and LC(90) values of second, third, and fourth instars of A. aegypti (LC(50) = 41.65, 139.51, 103.95; LC(90) = 117.81, 213.68, 367.82) and A. stephensi (LC(50) = 51.12, 105.52, 133.07; LC(90) = 134.81, 204.45, 285.35) were determined. At higher concentration (500 ppm), mortality starts within first 6 h of exposure. More than 50% mortality occurs within the first 24 h. The overall observed effects against A. aegypti and A. stephensi larvae after 48 h were increasing percent survival larvae, survival pupation, adult emergence with decreasing crude pigment extract concentration. These ensure that the resultant mosquito population reduction is substantial even where the larvicidal potential is minimal. The UV (λ (max) = 536 nm), TLC (Rf = 0.9), HPLC, and FTIR analysis of crude pigment shows the presence of prodigiosin as active compound. Thus, the active compound produced by this species would be more useful against vectors responsible for diseases of public health importance. This is the first report on mosquito larvicidal activity of prodigiosin produced by Serratia species.

  14. Effects of food, water depth, and temperature on diving activity of larval Anopheles gambiae sensu stricto: evidence for diving to forage.

    PubMed

    Phelan, Conan; Roitberg, Bernard D

    2013-12-01

    Anopheles gambiae larvae have frequently been observed to dive, but the ecology of this behavior has not been extensively examined. We manipulated food level, water depth, and temperature for individually-reared larvae and observed diving activity. Larvae dived more often under low food, which suggests that they dive to forage. There was only weak evidence for effects of water depth or temperature on diving. Experimental results are discussed in the context of energy budgets. Understanding larval ecology of this species is important for predicting how it will respond to environmental change. Further study is needed to assess the role that larval diving plays in both feeding ecology and thermal regulation of this and other medically important species.

  15. Nectar-related vs. human-related volatiles: behavioural response and choice by female and male Anopheles gambiae (Diptera: Culicidae) between emergence and first feeding.

    PubMed

    Foster, W A; Takken, W

    2004-04-01

    The close association of Anopheles gambiae Giles with humans and its females' ability to live on human blood alone suggest that females may ignore sources of sugar in favour of human blood as a source of energy. They have limited energy reserves at emergence, and at 27 degrees C both sexes generally die if they do not feed during night 1, 24-36 h after emergence. Food preferences during this critical period were tested by measuring responses to volatiles from honey and soiled socks, which served as surrogates for nectar-related and human-related volatiles in a wind-tunnel olfactometer. Both sexes responded more strongly to honey than to human volatiles, and given a choice, preferred honey over human volatiles. After 5 days of sugar access and maturation, males continued to prefer honey volatiles, whereas females changed behaviour, responding almost exclusively to human volatiles. Night 1 experiments also demonstrated that: (i). females previously having had sugar during the night of emergence responded more strongly to human volatiles; (ii). large-bodied mosquitoes of both sexes responded more strongly to honey than small-bodied ones; and (iii). females were equally responsive to honey in both early and late scotophase but were slightly more responsive to human volatiles in late scotophase. These results indicate that for a female's first meal, sugar is a viable option and is preferred when nectar-related stimuli are strong. This supports field evidence that sugar-feeding is a significant component of A. gambiae female behaviour.

  16. Diverged Alleles of the Anopheles gambiae Leucine-Rich Repeat Gene APL1A Display Distinct Protective Profiles against Plasmodium falciparum

    PubMed Central

    Mitri, Christian; Riehle, Michelle M.; Bischoff, Emmanuel; Brito-Fravallo, Emma; Takashima, Eizo; Thiery, Isabelle; Zettor, Agnes; Petres, Stephane; Bourgouin, Catherine; Vernick, Kenneth D.; Eiglmeier, Karin

    2012-01-01

    Functional studies have demonstrated a role for the Anopheles gambiae APL1A gene in resistance against the human malaria parasite, Plasmodium falciparum. Here, we exhaustively characterize the structure of the APL1 locus and show that three structurally different APL1A alleles segregate in the Ngousso colony. Genetic association combined with RNAi-mediated gene silencing revealed that APL1A alleles display distinct protective profiles against P. falciparum. One APL1A allele is sufficient to explain the protective phenotype of APL1A observed in silencing experiments. Epitope-tagged APL1A isoforms expressed in an in vitro hemocyte-like cell system showed that under assay conditions, the most protective APL1A isoform (APL1A2) localizes within large cytoplasmic vesicles, is not constitutively secreted, and forms only one protein complex, while a less protective isoform (APL1A1) is constitutively secreted in at least two protein complexes. The tested alleles are identical to natural variants in the wild A. gambiae population, suggesting that APL1A genetic variation could be a factor underlying natural heterogeneity of vector susceptibility to P. falciparum. PMID:23285147

  17. Diving behavior in Anopheles gambiae (Diptera: Culicidae): avoidance of a predacious wolf spider (Araneae: Lycosidae) in relation to life stage and water depth.

    PubMed

    Futami, Kyoko; Sonye, George; Akweywa, Peter; Kaneko, Satoshi; Minakawa, Noboru

    2008-11-01

    It has been suggested that mosquito larvae and pupae dive to avoid predators. We tested this predator-avoidance hypothesis by using immature Anopheles gambiae Giles (Diptera: Culicidae) and the wolf spider Pardosa messingerae (Stand) (Araneae: Lycosidae). Because previous studies have suggested that wolf spiders are poor predators of immature mosquitoes, we first examined the predatory ability of the wolf spider and found that the spider was effective at capturing all stages of larvae and pupae. The mortality from experimental cups containing deep water increased with the age of mosquitoes, with the exception of pupae. In contrast, this trend was not observed in shallow water. In particular, mortality was significantly lower in deep water during the second instar. During the third instar, the opposite trend was observed. When the effect of cannibalism was excluded by subtracting the number of missing mosquitoes for the treatment without spiders from those with spiders, the cannibalism corrected mortality was significantly lower in deep water during the second instar. The duration of diving by larvae and pupae decreased with age. With the exception of first instar, diving frequency also decreased with age. We postulate that this diving behavior allows An. gambiae to escape predation by wolf spiders, which supports the predator-avoidance hypothesis. This study indicates some important implications for vector control.

  18. Human Antibody Response to Anopheles gambiae Saliva: An Immuno-Epidemiological Biomarker to Evaluate the Efficacy of Insecticide-Treated Nets in Malaria Vector Control

    PubMed Central

    Drame, Papa M.; Poinsignon, Anne; Besnard, Patrick; Le Mire, Jacques; Dos-Santos, Maria A.; Sow, Cheikh S.; Cornelie, Sylvie; Foumane, Vincent; Toto, Jean-Claude; Sembene, Mbacké; Boulanger, Denis; Simondon, François; Fortes, Filomeno; Carnevale, Pierre; Remoue, Franck

    2010-01-01

    For the fight against malaria, the World Health Organization (WHO) has emphasized the need for indicators to evaluate the efficacy of vector-control strategies. This study investigates a potential immunological marker, based on human antibody responses to Anopheles saliva, as a new indicator to evaluate the efficacy of insecticide-treated nets (ITNs). Parasitological, entomological, and immunological assessments were carried out in children and adults from a malaria-endemic region of Angola before and after the introduction of ITNs. Immunoglobulin G (IgG) levels to An. gambiae saliva were positively associated with the intensity of An. gambiae exposure and malaria infection. A significant decrease in the anti-saliva IgG response was observed after the introduction of ITNs, and this was associated with a drop in parasite load. This study represents the first stage in the development of a new indicator to evaluate the efficacy of malaria vector-control strategies, which could apply in other arthropod vector-borne diseases. PMID:20595489

  19. Potential use of neem leaf slurry as a sustainable dry season management strategy to control the malaria vector Anopheles gambiae (DIPTERA: CULICIDAE) in west African villages.

    PubMed

    Luong, Kyphuong; Dunkel, Florence V; Coulibaly, Keriba; Beckage, Nancy E

    2012-11-01

    Larval management of the malaria vector, Anopheles gambiae Giles s.s., has been successful in reducing disease transmission. However, pesticides are not affordable to farmers in remote villages in Mali, and in other material resource poor countries. Insect resistance to insecticides and nontarget toxicity pose additional problems. Neem (Azadirachta indica A. Juss) is a tree with many beneficial, insect bioactive compounds, such as azadirachtin. We tested the hypothesis that neem leaf slurry is a sustainable, natural product, anopheline larvicide. A field study conducted in Sanambele (Mali) in 2010 demonstrated neem leaf slurry can work with only the available tools and resources in the village. Laboratory bioassays were conducted with third instar An. gambiae and village methods were used to prepare the leaf slurry. Experimental concentration ranges were 1,061-21,224 mg/L pulverized neem leaves in distilled water. The 50 and 90% lethal concentrations at 72 h were 8,825 mg/L and 15,212 mg/L, respectively. LC concentrations were higher than for other parts of the neem tree when compared with previous published studies because leaf slurry preparation was simplified by omitting removal of fibrous plant tissue. Using storytelling as a medium of knowledge transfer, villagers combined available resources to manage anopheline larvae. Preparation of neem leaf slurries is a sustainable approach which allows villagers to proactively reduce mosquito larval density within their community as part of an integrated management system. PMID:23270164

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

    PubMed

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

    PubMed Central

    2011-01-01

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

  2. Filaria vector competence of some Anopheles species.

    PubMed

    Zahedi, M; White, G B

    1994-03-01

    The filaria vector competence of Anopheles stephensi was compared with Brugia-susceptible Aedes aegypti Liverpool strain, An. gambiae Badagry Lagos strain and An. dirus Perlis Malaysia strain. An. stephensi ingested more Brugia pahangi microfilariae, had the highest infectivity rate and yielded more infective mosquitoes than the other two anopheline species. The overall vector competence of An. stephensi was 0.13 times that of Ae. aegypti, 0.62 times that of An. gambiae and 2.17 times that of An. dirus. However, heavy mortality among infected An. stephensi in the present investigation indicates that the filaria vectorial capacity of the mosquito might be limited epidemiologically. The relationship between filaria vector competence and mosquito foregut armature is discussed. It was observed that the relative vector competence of the three anopheline species tested was in the same order as their relative degrees of armature elaboration. The converse would be expected if foregut armatures really give partial protection to the mosquitoes against filarial infection. It is suggested that high host microfilariae density favours larval survival proportional to the degree of armature development in Anopheles (Cellia) species.

  3. Chemical constituents and larvicidal potential of Feronia limonia leaf essential oil against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus.

    PubMed

    Senthilkumar, A; Jayaraman, M; Venkatesalu, V

    2013-03-01

    In the present investigation, the leaf essential oil of Feronia limonia was evaluated for chemical constituents and mosquito larvicidal activity against the larvae of Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus. GC and GC-MS analyses revealed that the essential oil contain 51 compounds. Estragole (34.69 %) and β-pinene(23.59 %) were identified as the major constituents followed by methyl (Z)-caryophyllene (11.05 %), eugenol (6.50 %), linalool (3.97 %), phytol (3.27 %), sabinene (2.41 %) and limonene (2.27 %). Larval mortality was observed after 12 and 24 h of exposure period. The oil showed remarkable larvicidal activity against A. stephensi (LC(50) = 38.93 and LC(90) = 108.64 ppm (after 12 h); LC(50) = 15.03 and LC(90) = 36.69 ppm (after 24 h)), A. aegypti (LC(50) = 37.60 and LC(90) = 104.69 ppm (after 12 h); LC(50) = 11.59 and LC(90) = 42.95 ppm (after 24 h)) and C. quinquefasciatus (LC(50) = 52.08 and LC(90) = 124.33 ppm (after 12 h); LC(50) = 22.49 and LC(90) = 60.90 ppm (after 24 h)). Based on the results, the essential oil of F. limonia can be considered as a new source of larvicide for the control of vector mosquitoes.

  4. 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.

    PubMed

    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

  5. Trends in the selection of insecticide resistance in Anopheles gambiae s.l. mosquitoes in northwest Tanzania during a community randomized trial of longlasting insecticidal nets and indoor residual spraying.

    PubMed

    Matowo, J; Kitau, J; Kaaya, R; Kavishe, R; Wright, A; Kisinza, W; Kleinschmidt, I; Mosha, F; Rowland, M; Protopopoff, N

    2015-03-01

    Anopheles gambiae s.l. (Diptera: Culicidae) in Muleba, Tanzania has developed high levels of resistance to most insecticides currently advocated for malaria control. The kdr mutation has almost reached fixation in An. gambiae s.s. in Muleba. This change has the potential to jeopardize malaria control interventions carried out in the region. Trends in insecticide resistance were monitored in two intervention villages using World Health Organization (WHO) susceptibility test kits. Additional mechanisms contributing to observed phenotypic resistance were investigated using Centers for Disease Control (CDC) bottle bioassays with piperonylbutoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) synergists. Resistance genotyping for kdr and Ace-1 alleles was conducted using quantitative polymerase chain reaction (qPCR). In both study villages, high phenotypic resistance to several pyrethroids and DDT was observed, with mortality in the range of 12-23%. There was a sharp decrease in mortality in An. gambiae s.l. exposed to bendiocarb (carbamate) from 84% in November 2011 to 31% in December 2012 after two rounds of bendiocarb-based indoor residual spraying (IRS). Anopheles gambiae s.l. remained susceptible to pirimiphos-methyl (organophosphate). Bendiocarb-based IRS did not lead to the reversion of pyrethroid resistance. There was no evidence for selection for Ace-1 resistance alleles. The need to investigate the operational impact of the observed resistance selection on the effectiveness of longlasting insecticidal nets and IRS for malaria control is urgent. PMID:25537754

  6. Trends in the selection of insecticide resistance in Anopheles gambiae s.l. mosquitoes in northwest Tanzania during a community randomized trial of longlasting insecticidal nets and indoor residual spraying.

    PubMed

    Matowo, J; Kitau, J; Kaaya, R; Kavishe, R; Wright, A; Kisinza, W; Kleinschmidt, I; Mosha, F; Rowland, M; Protopopoff, N

    2015-03-01

    Anopheles gambiae s.l. (Diptera: Culicidae) in Muleba, Tanzania has developed high levels of resistance to most insecticides currently advocated for malaria control. The kdr mutation has almost reached fixation in An. gambiae s.s. in Muleba. This change has the potential to jeopardize malaria control interventions carried out in the region. Trends in insecticide resistance were monitored in two intervention villages using World Health Organization (WHO) susceptibility test kits. Additional mechanisms contributing to observed phenotypic resistance were investigated using Centers for Disease Control (CDC) bottle bioassays with piperonylbutoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) synergists. Resistance genotyping for kdr and Ace-1 alleles was conducted using quantitative polymerase chain reaction (qPCR). In both study villages, high phenotypic resistance to several pyrethroids and DDT was observed, with mortality in the range of 12-23%. There was a sharp decrease in mortality in An. gambiae s.l. exposed to bendiocarb (carbamate) from 84% in November 2011 to 31% in December 2012 after two rounds of bendiocarb-based indoor residual spraying (IRS). Anopheles gambiae s.l. remained susceptible to pirimiphos-methyl (organophosphate). Bendiocarb-based IRS did not lead to the reversion of pyrethroid resistance. There was no evidence for selection for Ace-1 resistance alleles. The need to investigate the operational impact of the observed resistance selection on the effectiveness of longlasting insecticidal nets and IRS for malaria control is urgent.

  7. Experimental Swap of Anopheles gambiae's Assortative Mating Preferences Demonstrates Key Role of X-Chromosome Divergence Island in Incipient Sympatric Speciation

    PubMed Central

    Aboagye-Antwi, Fred; Alhafez, Nahla; Weedall, Gareth D.; Brothwood, Jessica; Kandola, Sharanjit; Paton, Doug; Fofana, Abrahamane; Olohan, Lisa; Betancourth, Mauro Pazmiño; Ekechukwu, Nkiru E.; Baeshen, Rowida; Traorè, Sékou F.; Diabate, Abdoulaye; Tripet, Frédéric

    2015-01-01

    Although many theoretical models of sympatric speciation propose that genes responsible for assortative mating amongst incipient species should be associated with genomic regions protected from recombination, there are few data to support this theory. The malaria mosquito, Anopheles gambiae, is known for its sympatric cryptic species maintained by pre-mating reproductive isolation and its putative genomic islands of speciation, and is therefore an ideal model system for studying the genomic signature associated with incipient sympatric speciation. Here we selectively introgressed the island of divergence located in the pericentric region of the X chromosome of An. gambiae s.s. into its sister taxon An. coluzzii through 5 generations of backcrossing followed by two generations of crosses within the introgressed strains that resulted in An. coluzzii-like recombinant strains fixed for the M and S marker in the X chromosome island. The mating preference of recombinant strains was then tested by giving virgin recombinant individuals a choice of mates with X-islands matching and non-matching their own island type. We show through genetic analyses of transferred sperm that recombinant females consistently mated with matching island-type males thereby associating assortative mating genes with the X-island of divergence. Furthermore, full-genome sequencing confirmed that protein-coding differences between recombinant strains were limited to the experimentally swapped pericentromeric region. Finally, targeted-genome comparisons showed that a number of these unique differences were conserved in sympatric field populations, thereby revealing candidate speciation genes. The functional demonstration of a close association between speciation genes and the X-island of differentiation lends unprecedented support to island-of-speciation models of sympatric speciation facilitated by pericentric recombination suppression. PMID:25880677

  8. A Small-Scale Field Trial of Pyriproxyfen-Impregnated Bed Nets against Pyrethroid-Resistant Anopheles gambiae s.s. in Western Kenya

    PubMed Central

    Kawada, Hitoshi; Dida, Gabriel O.; Ohashi, Kazunori; Kawashima, Emiko; Sonye, George; Njenga, Sammy M.; Mwandawiro, Charles; Minakawa, Noboru

    2014-01-01

    Pyrethroid resistance is becoming a major problem for vector control programs, because at present, there are few suitable chemical substitutes for pyrethroids, as when used on bed nets the insecticide must have low mammalian toxicity as well as high activity to mosquitoes. Pyriproxyfen (PPF) is one of the most active chemicals among the juvenile hormone mimic (JHM) group. Sterilizing mosquitoes by using PPF could be a potential control measure for pyrethroid-resistant malaria vectors. We investigated the sterilizing effects of two types of PPF-impregnated bed nets – a 1% PPF-impregnated net and a 1% PPF +2% permethrin-impregnated net (Olyset Duo) – to pyrethroid-resistant wild population of Anopheles gambiae s.s. in western Kenya. High mortality of blood-fed mosquitos was observed 3 days post-collection, in the houses where PPF-impregnated nets were used, indicating the effect of PPF on the longevity of mosquitos that came in contact with the net. Reduction in the number of ovipositing females, number of eggs, and number of progeny per female were also observed in the houses in which both Olyset Duo and PPF-impregnated nets were used. This is the first field study showing the high sterilizing efficacy of PPF against wild pyrethroid-resistant An. gambiae s.s. population. In addition, we recognized the necessity of combined use of permethrin with PPF, in order to reduce the risk of mosquito bites and provide a level of personal protection. Further studies on wild pyrethroid-resistant mosquito populations such as An. arabiensis and An. funestus s.s. would provide more information on the practical use of the PPF-impregnated bed nets. PMID:25333785

  9. Parallel evolution or purifying selection, not introgression, explains similarity in the pyrethroid detoxification linked GSTE4 of Anopheles gambiae and An. arabiensis.

    PubMed

    Wilding, C S; Weetman, D; Rippon, E J; Steen, K; Mawejje, H D; Barsukov, I; Donnelly, M J

    2015-02-01

    Insecticide resistance is a major impediment to the control of vectors and pests of public health importance and is a strongly selected trait capable of rapid spread, sometimes even between closely related species. Elucidating the mechanisms generating insecticide resistance in mosquito vectors of disease, and understanding the spread of resistance within and between populations and species are vital for the development of robust resistance management strategies. Here, we studied the mechanisms of resistance in two sympatric members of the Anopheles gambiae species complex-the major vector of malaria in sub-Saharan Africa-to understand how resistance has developed and spread in eastern Uganda, a region with some of the highest levels of malaria. In eastern Uganda, where the mosquitoes Anopheles arabiensis and An. gambiae can be found sympatrically, low levels of hybrids (0.4 %) occur, offering a route for introgression of adaptively important variants between species. In independent microarray studies of insecticide resistance, Gste4, an insect-specific glutathione S-transferase, was among the most significantly up-regulated genes in both species. To test the hypothesis of interspecific introgression, we sequenced 2.3 kbp encompassing Gste4. Whilst this detailed sequencing ruled out introgression, we detected strong positive selection acting on Gste4. However, these sequences, followed by haplotype-specific qPCR, showed that the apparent up-regulation in An. arabiensis is a result of allelic variation across the microarray probe binding sites which artefactually elevates the gene expression signal. Thus, face-value acceptance of microarray data can be misleading and it is advisable to conduct a more detailed investigation of the causes and nature of such signal. The identification of positive selection acting on this locus led us to functionally express and characterise allelic variants of GSTE4. Although the in vitro data do not support a direct role for GSTE4

  10. 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).

    PubMed

    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

  11. Differential Induction of Proteins in Anopheles gambiae sensu stricto (Diptera: Cullicidae) Larvae in Response to Heavy Metal Selection

    PubMed Central

    Mireji, Paul O.; Keating, Joseph; Kenya, Eucharia; Mbogo, Charles; Nyambaka, Hudson; Osir, Ellie; Githure, John; Beier, John

    2009-01-01

    Investigations were conducted to establish the magnitude and pattern of differential expression of proteins due to generational selection of third instar An. gambiae s.s. larvae by cadmium, copper and lead heavy metals, three possible common urban pollutants. A susceptible strain of An. gambiae s.s. third instar larvae was separately placed under selection pressure with cadmium, copper and lead at LC30 and controls through five generations. First, third and fifth generation selection survivors were screened for differentially expressed proteins relative to non-exposed control by two-dimensional gel electrophoresis. Distribution patterns of the spots were analysed by Chi Square or Fishers exact test and variations in expressions between and within generation by ANOVA. Most differentially expressed spots were acidic and of low molecular weight among all metals and generations. Type of heavy metals and generation were main indicators of variations in differential expressions. Variation between generations was most significant among cadmium-selected populations of which most number of spots were induced in the fifth generation. Most spots were induced in the copper-selected population in the third generation. The induced protein spots may be products from respective genes that respond to heavy metals and counter their toxicity, thus building An. gambiae s.s. tolerance to these pollutants. The differential pattern and magnitude of expressed spots has potential application as molecular markers for assessment of anopheline adaptation status to heavy metals, and provide insight into the extent of environmental pollution. PMID:20651951

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

    PubMed Central

    2012-01-01

    Background The purpose of this study was to determine whether the sugar-or-blood meal choice of Anopheles gambiae females one day after emergence is influenced by blood-host presence and accessibility, nectariferous plant abundance, and female size. This tested the hypothesis that the initial meal of female An. gambiae is sugar, even when a blood host is available throughout the night, and, if not, whether the use of a bed net diverts mosquitoes to sugar sources. Methods Females and males <1-day post-emergence were released in a mesocosm. Overnight they had access to either one or six Senna didymobotrya plants. Simultaneously they had access to a human blood host, either for 8 h or for only 30 min at dusk and dawn (the remainder of the night being excluded by an untreated bed net). In a third situation, the blood host was not present. All mosquitoes were collected in the morning. Their wing lengths, an indicator of pre-meal energetic state, were measured, and their meal choice was determined by the presence of midgut blood and of fructose. Results Female sugar feeding after emergence was facultative. When a blood host was accessible for 8 h per night, 92% contained blood, and only 3.7% contained sugar. Even with the use of a bed net, 78% managed to obtain a blood meal during the 30 min of accessibility at dusk or dawn, but 14% of females were now fructose-positive. In the absence of a blood host, and when either one or six plants were available, a total of 21.7% and 23.6% of females and 30.8% and 43.5% of males contained fructose, respectively. Feeding on both sugar and blood was more likely with bed net use and with greater plant abundance. Further, mosquitoes that fed on both resources were more often small and had taken a sugar meal earlier than the blood meal. The abundance of sugar hosts also affected the probability of sugar feeding by males and the amount of fructose obtained by both males and females. Conclusion Even in an abundance of potential sugar

  13. Insect repellent activity of medicinal plant oils against Aedes aegypti (Linn.), Anopheles minimus (Theobald) and Culex quinquefasciatus Say based on protection time and biting rate.

    PubMed

    Phasomkusolsil, Siriporn; Soonwera, Mayura

    2010-07-01

    This study investigated insect bite protection and length of the protection with 30 repellents which were divided into 3 categories: plant oil, essential oil and essential oil with ethyl alcohol, tested against three mosquito species, Aedes aegypti, Anopheles minimus and Culex quinquefasciatus, under laboratory conditions. The plant oil group was comprised of Phlai (Zingiber cassumunar) and Sweet basil (Ocimum basilicum). Both substances were effective as repellents and feeding deterrents against An. minimus (205 minutes protection time and a biting rate of 0.9%), Cx. quinquefasciatus (165 minutes protection time and 0.9% biting rate) and Ae. aegypti (90 minutes protection time and 0.8% biting rate). Essential oil from citronella grass (Cymbopogon nardus) exhibited protection against biting from all 3 mosquito species: for An. minimus, Cx. quinquefasciatus and Ae. aegypti, the results were 130 minutes and 0.9%, 140 minutes and 0.8%, and 115 minutes and 0.8%, respectively. The period of protection time against Ae. aegypti for all repellent candidates tested was lower than the Thai Industrial Standards Institute (TISI) determined time of greater than 2 hours. PMID:21073057

  14. Effect of Cymbopogon citratus (lemongrass) and Syzygium aromaticum (clove) oils on the morphology and mortality of Aedes aegypti and Anopheles dirus larvae.

    PubMed

    Soonwera, Mayura; Phasomkusolsil, Siriporn

    2016-04-01

    Cymbopogon citratus (lemongrass) and Syzygium aromaticum (clove) oils were evaluated to determine mortality rates, morphological aberrations, and persistence when used against third and fourth larval instars of Aedes aegypti and Anopheles dirus. The oils were evaluated at 1, 5, and 10 % concentrations in mixtures with soybean oil. Persistence of higher concentrations was measured over a period of 10 days. For Ae. aegypti, both plant oils caused various morphological aberrations to include deformed larvae, incomplete eclosion, white pupae, deformed pupae, dead normal pupae, and incomplete pupal eclosion. All of these aberrations led to larval mortality. In Ae. aegypti larvae, there were no significant differences in mortality at days 1, 5, and 10 or between third and fourth larval instar exposure. In An. dirus, morphological aberrations were rare and S. aromaticum oil was more effective in causing mortality among all larval stages. Both oils were equally effective at producing mortality on days 1, 5, and 10. Both oils had slightly increased LT50 rates from day 1 to day 10. In conclusion, both lemongrass and clove oils have significant effects on the immature stages of Ae. aegypti and An. dirus and could potentially be developed for use as larvicides. PMID:26796022

  15. Effect of Cymbopogon citratus (lemongrass) and Syzygium aromaticum (clove) oils on the morphology and mortality of Aedes aegypti and Anopheles dirus larvae.

    PubMed

    Soonwera, Mayura; Phasomkusolsil, Siriporn

    2016-04-01

    Cymbopogon citratus (lemongrass) and Syzygium aromaticum (clove) oils were evaluated to determine mortality rates, morphological aberrations, and persistence when used against third and fourth larval instars of Aedes aegypti and Anopheles dirus. The oils were evaluated at 1, 5, and 10 % concentrations in mixtures with soybean oil. Persistence of higher concentrations was measured over a period of 10 days. For Ae. aegypti, both plant oils caused various morphological aberrations to include deformed larvae, incomplete eclosion, white pupae, deformed pupae, dead normal pupae, and incomplete pupal eclosion. All of these aberrations led to larval mortality. In Ae. aegypti larvae, there were no significant differences in mortality at days 1, 5, and 10 or between third and fourth larval instar exposure. In An. dirus, morphological aberrations were rare and S. aromaticum oil was more effective in causing mortality among all larval stages. Both oils were equally effective at producing mortality on days 1, 5, and 10. Both oils had slightly increased LT50 rates from day 1 to day 10. In conclusion, both lemongrass and clove oils have significant effects on the immature stages of Ae. aegypti and An. dirus and could potentially be developed for use as larvicides.

  16. The effects of herbal essential oils on the oviposition-deterrent and ovicidal activities of Aedes aegypti (Linn.), Anopheles dirus (Peyton and Harrison) and Culex quinquefasciatus (Say).

    PubMed

    Siriporn, P; Mayura, S

    2012-03-01

    The effect of oviposition-deterrent and ovicidal of seven essential oils were evaluated towards three mosquito vectors, Aedes aegypti, Anopheles dirus and Culex quinquefasciatus. The oviposition activity index (OAI) values of six essential oils namely Cananga odorata, Cymbopogon citratus, Cymbopogon nardus, Eucalyptus citriodora, Ocimum basilicum and Syzygium aromaticum indicated that there were more deterrent than the control whereas Citrus sinensis oil acted as oviposition attractant. At higher concentration (10%) of Ca. odorata (ylang ylang flowers) showed high percent effective repellency (ER) against oviposition at 99.4% to Ae. aegypti, 97.1% to An. dirus and 100% to Cx. quinquefasciatus, respectively. The results showed that mean numbers of eggs were lower in treated than in untreated water. In addition, there was an inverse relationship between essential oil concentrations and ovicidal activity. As the concentration of essential oil increased from 1%, 5% and up to 10% conc., the hatching rate decreased. The essential oil of Ca. odorata at 10% conc. gave minimum egg hatch of 10.4% (for Ae. aegypti), 0.8% (for An. dirus) and 1.1% (for Cx. quinquefasciatus) respectively. These results clearly revealed that the essential oil of Ca. odorata served as a potential oviposition-deterrent and ovicidal activity against Ae. aegypti, An. dirus and Cx. quinquefasciatus. PMID:22543614

  17. The effect of water turbidity on the near-surface water temperature of larval habitats of the malaria mosquito Anopheles gambiae

    NASA Astrophysics Data System (ADS)

    Paaijmans, K. P.; Takken, W.; Githeko, A. K.; Jacobs, A. F. G.

    2008-11-01

    Water temperature is an important determinant in many aquatic biological processes, including the growth and development of malaria mosquito ( Anopheles arabiensis and A. gambiae) immatures. Water turbidity affects water temperature, as suspended particles in a water column absorb and scatter sunlight and hence determine the extinction of solar radiation. To get a better understanding of the relationship between water turbidity and water temperature, a series of semi-natural larval habitats (diameter 0.32 m, water depth 0.16 m) with increasing water turbidity was created. Here we show that at midday (1300 hours) the upper water layer (thickness of 10 mm) of the water pool with the highest turbidity was on average 2.8°C warmer than the same layer of the clearest water pool. Suspended soil particles increase the water temperature and furthermore change the temperature dynamics of small water collections during daytime, exposing malaria mosquito larvae, which live in the top water layer, longer to higher temperatures.

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

    PubMed

    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. Comparative structural and functional analysis of the larval and adult dorsal vessel and its role in hemolymph circulation in the mosquito Anopheles gambiae

    PubMed Central

    League, Garrett P.; Onuh, Ogechukwu C.; Hillyer, Julián F.

    2015-01-01

    Hemolymph circulation in insects is driven primarily by the contractile action of a dorsal vessel, which is divided into an abdominal heart and a thoracic aorta. As holometabolous insects, mosquitoes undergo striking morphological and physiological changes during metamorphosis. This study presents a comprehensive structural and functional analysis of the larval and adult dorsal vessel in the malaria mosquito Anopheles gambiae. Using intravital video imaging we show that, unlike the adult heart, the larval heart contracts exclusively in the anterograde direction and does not undergo heartbeat directional reversals. The larval heart contracts 24% slower than the adult heart, and hemolymph travels across the larval dorsal vessel at a velocity that is 68% slower than what is seen in adults. By fluorescently labeling muscle tissue we show that although the general structure of the heart and its ostia are similar across life stages, the heart-associated alary muscles are significantly less robust in larvae. Furthermore, unlike the adult ostia, which are the entry points for hemolymph into the heart, the larval ostia are almost entirely lacking in incurrent function. Instead, hemolymph enters the larval heart through incurrent openings located at the posterior terminus of the heart. These posterior openings are structurally similar across life stages, but in adults have an opposite, excurrent function. Finally, the larval aorta and heart differ significantly in the arrangement of their cardiomyocytes. In summary, this study provides an in-depth developmental comparison of the circulatory system of larval and adult mosquitoes. PMID:25524976

  20. A determinant of odorant specificity is located at the extracellular loop 2-transmembrane domain 4 interface of an Anopheles gambiae odorant receptor subunit.

    PubMed

    Hughes, David T; Wang, Guirong; Zwiebel, Laurence J; Luetje, Charles W

    2014-11-01

    To explore the structural basis for odorant specificity in odorant receptors of the human malaria vector mosquito, Anopheles gambiae, odorant-binding subunits (Agam\\Ors) expressed in Xenopus oocytes in combination with Agam\\Orco (coreceptor subunit) were assayed by 2-electrode voltage clamp against 25 structurally related odorants. Agam\\Or13 and Agam\\Or15 display 82% amino acid identity and had similar, but somewhat distinct odorant response profiles. The ratio of acetophenone to 4-methylphenol responses was used in a mutation-based analysis of Agam\\Or15, interchanging 37 disparate residues between Agam\\Or15 and Agam\\Or13. Eleven mutations caused significant changes in odorant responsiveness. Mutation of alanine 195 resulted in the largest shift in response ratio from Agam\\Or15 toward Agam\\Or13. Concentration-response analysis for a series of mutations of residue 195 revealed a large effect on acetophenone sensitivity, with EC50 values varying by >1800-fold and correlating with residue side chain length. Similar results were obtained for propiophenone and benzaldehyde. But, for other odorants, such as 4-methylphenol, 4-methylbenzaldehyde, and 4-methylpropiophenone, the effect of mutation was much smaller (EC50 values varied by ≤16-fold). These results show that alanine 195, putatively located at the second extracellular loop/fourth transmembrane domain interface, plays a critical role in determining the odorant response specificity of Agam\\Or15. PMID:25270378

  1. Infrared video tracking of Anopheles gambiae at insecticide-treated bed nets reveals rapid decisive impact after brief localised net contact

    PubMed Central

    Parker, Josephine E.A.; Angarita-Jaimes, Natalia; Abe, Mayumi; Towers, Catherine E.; Towers, David; McCall, Philip J.

    2015-01-01

    Long-lasting insecticidal bed nets (LLINs) protect humans from malaria transmission and are fundamental to malaria control worldwide, but little is known of how mosquitoes interact with nets. Elucidating LLIN mode of action is essential to maintain or improve efficacy, an urgent need as emerging insecticide resistance threatens their future. Tracking multiple free-flying Anopheles gambiae responding to human-occupied bed nets in a novel large-scale system, we characterised key behaviours and events. Four behavioural modes with different levels of net contact were defined: swooping, visiting, bouncing and resting. Approximately 75% of all activity occurred at the bed net roof where multiple brief contacts were focussed above the occupant’s torso. Total flight and net contact times were lower at LLINs than untreated nets but the essential character of the response was unaltered. LLINs did not repel mosquitoes but impacted rapidly: LLIN contact of less than 1 minute per mosquito during the first ten minutes reduced subsequent activity; after thirty minutes, activity at LLINs was negligible. Velocity measurements showed that mosquitoes detected nets, including unbaited untreated nets, prior to contact. This is the most complete characterisation of mosquito-LLIN interactions to date, and reveals many aspects of LLIN mode of action, important for developing the next generation of LLINs. PMID:26323965

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

    PubMed

    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

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

    PubMed

    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

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

    PubMed

    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

  5. Identification of chemical constituents and larvicidal activity of essential oil from Murraya exotica L. (Rutaceae) against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Krishnamoorthy, Shanmugam; Chandrasekaran, Manivachagam; Raj, Gnanaprakasam Adaikala; Jayaraman, Mahalingam; Venkatesalu, Venugopalan

    2015-05-01

    This study was conducted to evaluate the phytochemical composition and larvicidal effect of leaf essential oil from Murraya exotica against early fourth-instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. Gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS) analyses revealed that the essential oil contained 27 components. The major chemical components identified were β-humulene (40.62%), benzyl benzoate (23.96%), β-caryophyllene (7.05%) and α-terpinene (5.66%). The larval mortality was observed after 12 and 24 h of exposure period. The results revealed that essential oil showed varied levels of larvicidal activity against A. aegypti, A. stephensi and C. quinquefasciatus. After 12 h of exposure period, the larvicidal activities were LC₅₀ = 74.7 and LC₉₀ = 152.7 ppm (A. aegypti), LC₅₀ = 56.3 and LC₉₀ = 107.8 ppm (A. stephensi ), and LC₅₀ = 74.4 and LC₉₀ = 136.9 ppm (C. quinquefasciatus) and the larvicidal activities after 24 h of exposure period were LC₅₀ = 35.8 and LC₉₀ = 85.4 ppm (A. aegypti), LC₅₀ = 31.3 and LC₉₀ = 75.1 ppm (A. stephensi), and LC₅₀ = 43.2 and LC₉₀ = 103.2 ppm (C. quinquefasciatus). These results suggest that leaf essential oil from M. exotica is a promising and eco-friendly source of natural larvicidal agent against A. aegypti, A. stephensi and C. quinquefasciatus. PMID:25697880

  6. Identification of chemical constituents and larvicidal activity of essential oil from Murraya exotica L. (Rutaceae) against Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Krishnamoorthy, Shanmugam; Chandrasekaran, Manivachagam; Raj, Gnanaprakasam Adaikala; Jayaraman, Mahalingam; Venkatesalu, Venugopalan

    2015-05-01

    This study was conducted to evaluate the phytochemical composition and larvicidal effect of leaf essential oil from Murraya exotica against early fourth-instar larvae of Aedes aegypti, Anopheles stephensi and Culex quinquefasciatus. Gas chromatography (GC) and gas chromatography mass spectrometry (GC-MS) analyses revealed that the essential oil contained 27 components. The major chemical components identified were β-humulene (40.62%), benzyl benzoate (23.96%), β-caryophyllene (7.05%) and α-terpinene (5.66%). The larval mortality was observed after 12 and 24 h of exposure period. The results revealed that essential oil showed varied levels of larvicidal activity against A. aegypti, A. stephensi and C. quinquefasciatus. After 12 h of exposure period, the larvicidal activities were LC₅₀ = 74.7 and LC₉₀ = 152.7 ppm (A. aegypti), LC₅₀ = 56.3 and LC₉₀ = 107.8 ppm (A. stephensi ), and LC₅₀ = 74.4 and LC₉₀ = 136.9 ppm (C. quinquefasciatus) and the larvicidal activities after 24 h of exposure period were LC₅₀ = 35.8 and LC₉₀ = 85.4 ppm (A. aegypti), LC₅₀ = 31.3 and LC₉₀ = 75.1 ppm (A. stephensi), and LC₅₀ = 43.2 and LC₉₀ = 103.2 ppm (C. quinquefasciatus). These results suggest that leaf essential oil from M. exotica is a promising and eco-friendly source of natural larvicidal agent against A. aegypti, A. stephensi and C. quinquefasciatus.

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

    PubMed

    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.

  8. Strong larvicidal potential of Artemisia annua leaf extract against malaria (Anopheles stephensi Liston) and dengue (Aedes aegypti L.) vectors and bioassay-driven isolation of the marker compounds.

    PubMed

    Sharma, Gaurav; Kapoor, Himanshi; Chopra, Madhu; Kumar, Kaushal; Agrawal, Veena

    2014-01-01

    Malaria and dengue are the two most important vector-borne human diseases caused by mosquito vectors Anopheles stephensi and Aedes aegypti, respectively. Of the various strategies adopted for eliminating these diseases, controlling of vectors through herbs has been reckoned as one of the important measures for preventing their resurgence. Artemisia annua leaf chloroform extract when tried against larvae of A. stephensi and A. aegypti has shown a strong larvicidal activity against both of these vectors, their respective LC50 and LC90 values being 0.84 and 4.91 ppm for A. stephensi and 0.67 and 5.84 ppm for A. aegypti. The crude extract when separated through column chromatography using petroleum ether-ethyl acetate gradient (0-100%) yielded 76 fractions which were pooled into three different active fractions A, B and C on the basis of same or nearly similar R f values. The aforesaid pooled fractions when assayed against the larvae of A. stephensi too reported a strong larvicidal activity. The respective marker compound purified from the individual fractions A, B and C, were Artemisinin, Arteannuin B and Artemisinic acid, as confirmed and characterized through FT-IR and NMR. This is our first report of strong mortality of A. annua leaf chloroform extract against vectors of two deadly diseases. This technology can be scaled up for commercial exploitation. PMID:24158647

  9. Phytochemical profile and larvicidal properties of seed essential oil from Nigella sativa L. (Ranunculaceae), against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Raj, Gnanaprakasam Adaikala; Chandrasekaran, Manivachagam; Krishnamoorthy, Shanmugam; Jayaraman, Mahalingam; Venkatesalu, Venugopalan

    2015-09-01

    The present study deals with investigation of larvicidal activity and their chemical constituents of the essential oil from the seeds of Nigella sativa L. (Ranunculaceae). Totally, 18 chemical compounds were identified by GC and GC-MS analysis. Thymol (19.13 %) and α-phellandrene (14.9 %) were identified as major chemical components followed by camphor (12.14 %), borneol (11.31 %), and carvacrol (8.65 %). The larval mortality was observed after 12 and 24 h of exposure period. The results revealed that the essential oil were evaluated against the fourth instar larvae of Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. After 12 h of exposure period, the larvicidal activities were LC50 = 196.9 and LC90 = 523.5 ppm (A. aegypti), LC50 = 88.1 and LC90 = 272.4 ppm (A.stephensi), and LC50 = 241.6 and LC90 = 545.4 ppm (C. quinquefasciatus), and the larvicidal activities after 24 h of exposure period were LC50 = 99.9 and LC90 = 300.8 ppm (A. aegypti), LC50 = 53.9 and LC90 = 172.6 ppm (A. stephensi), and LC50 = 141.7 and LC90 = 364.0 ppm (C. quinquefasciatus). The results of the present study showed that the essential oil from seeds of N. sativa is inexpensive food formulation and new source of natural larvicidal agent. PMID:26091760

  10. Phytochemical profile and larvicidal properties of seed essential oil from Nigella sativa L. (Ranunculaceae), against Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Raj, Gnanaprakasam Adaikala; Chandrasekaran, Manivachagam; Krishnamoorthy, Shanmugam; Jayaraman, Mahalingam; Venkatesalu, Venugopalan

    2015-09-01

    The present study deals with investigation of larvicidal activity and their chemical constituents of the essential oil from the seeds of Nigella sativa L. (Ranunculaceae). Totally, 18 chemical compounds were identified by GC and GC-MS analysis. Thymol (19.13 %) and α-phellandrene (14.9 %) were identified as major chemical components followed by camphor (12.14 %), borneol (11.31 %), and carvacrol (8.65 %). The larval mortality was observed after 12 and 24 h of exposure period. The results revealed that the essential oil were evaluated against the fourth instar larvae of Aedes aegypti, Anopheles stephensi, and Culex quinquefasciatus. After 12 h of exposure period, the larvicidal activities were LC50 = 196.9 and LC90 = 523.5 ppm (A. aegypti), LC50 = 88.1 and LC90 = 272.4 ppm (A.stephensi), and LC50 = 241.6 and LC90 = 545.4 ppm (C. quinquefasciatus), and the larvicidal activities after 24 h of exposure period were LC50 = 99.9 and LC90 = 300.8 ppm (A. aegypti), LC50 = 53.9 and LC90 = 172.6 ppm (A. stephensi), and LC50 = 141.7 and LC90 = 364.0 ppm (C. quinquefasciatus). The results of the present study showed that the essential oil from seeds of N. sativa is inexpensive food formulation and new source of natural larvicidal agent.

  11. Insecticidal potential of Ocimum canum plant extracts against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus larval and adult mosquitoes (Diptera: Culicidae).

    PubMed

    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.

  12. Contribution of Anopheles funestus, An. gambiae and An. nili (Diptera: Culicidae) to the perennial malaria transmission in the southern and western forest areas of Côte d’Ivoire

    PubMed Central

    Adja, A M; N’goran, E K; Koudou, B G; Dia, I; Kengne, P; Fontenille, D; Chandre, F

    2011-01-01

    The involvement of members of the Anopheles gambiae complex Giles and An. funestus Giles and An. nili Theobald groups in the transmission of Plasmodium falciparum was recently investigated in the villages of Gbatta and Kpéhiri, which lie, respectively, in forest areas in the west and south of Côte d’Ivoire. Adult female mosquitoes were collected, using human landing catches, inside and outside dwellings. After identification and dissection, the heads and thoraces of all the anopheline mosquitoes were tested, in an ELISA, for circumsporozoite protein (CSP). All the female anopheline mosquitoes collected and identified to species using PCR were found to be An. gambiae s.s., An. nili s.s. or An. funestus s.s., with An. gambiae s.s. and An. funestus s.s. predominant in Gbatta but An. nili s.s. the most common species in Kpéhiri. In Gbatta, 3.1% of the female An. gambiae collected, 5.0% of the female An. funestus and 1.8% of the female An. nili were found CSP-positive. The corresponding values in Kpéhiri were even higher, at 5.9%, 6.2% and 2.4%, respectively. The estimated entomological inoculation rates (EIR) were very high: 302 infected bites (139 from An. gambiae, 146 from An. funestus and 17 from An. nili)/person-year in Gbatta and 484 infected bites (204 from An. gambiae, 70 from An. funestus and 210 from An. nili)/person-year in Kpéhiri. In Gbatta, An. gambiae s.s. was responsible for most of the rainy-season transmission while An. funestus became the main malaria vector in the dry seasons. In Kpéhiri, however, An. nili appeared to be the main vector throughout the year, with An. gambiae of secondary importance and An. funestus only becoming a significant vector during the rainy season. Although, in both study sites, intense transmission was therefore occurring and the same three species of anopheline mosquito were present, the relative importance of each mosquito species in the epidemiology of the human malaria at each site differed markedly. PMID

  13. Variation in exposure to Anopheles gambiae salivary gland peptide (gSG6-P1) across different malaria transmission settings in the western Kenya highlands

    PubMed Central

    2012-01-01

    Background The existing metrics of malaria transmission are limited in sensitivity under low transmission intensity. Robust surveillance systems are needed as interventions to monitor reduced transmission and prevention of rapid reintroduction. Serological tools based on antibody responses to parasite and vector antigens are potential tools for transmission measurements. The current study sought to evaluate antibody responses to Anopheles gambiae salivary gland peptide (gSG6- P1), as a biomarker of human exposure to Anopheles bites, in different transmission settings and seasons. The comparison between anti-MSP-119 IgG immune responders and non-responders allowed exploring the robustness of the gSG6-P1 peptide as a surveillance tool in an area of decreasing malaria transmission. Methods Total IgG levels to gSG6-P1 were measured in an age-stratified cohort (< 5, 5–14 and ≥ 15 years) in a total of 1,366 participants from three localities in western Kenya [Kisii (hypoendemic), Kakamega (mesoendemic), and Kombewa (hyperendemic)] including 607 sera that were additionally tested for MSP-119 specific responses during a low and a high malaria transmission seasons. Antibody prevalence and levels were compared between localities with different transmission intensities. Regression analysis was performed to examine the association between gSG6-P1 and MSP-119 seroprevalence and parasite prevalence. Result Seroprevalence of gSG6-P1 in the uphill population was 36% while it was 50% valley bottom (χ2 = 13.2, df = 1, p < 0.001). Median gSG6-P1 antibody levels in the Valley bottom were twice as high as that observed in the uphill population [4.50 vs. 2.05, p < 0.001] and showed seasonal variation. The odds of gSG6-P1 seropositives having MSP-119 antibodies were almost three times higher than the odds of seronegatives (OR = 2.87, 95% CI [1.977, 4.176]). The observed parasite prevalence for Kisii, Kakamega and Kombewa were 4%, 19.7% and 44.6% whilst the

  14. A field attempt to assess the mating competitiveness of sterile males produced by crossing 2 members of the Anopheles gambiae complex

    PubMed Central

    Davidson, G.; Odetoyinbo, J. A.; Colussa, B.; Coz, J.

    1970-01-01

    In laboratory experiments sterile males produced by crossing member species of the Anopheles gambiae complex competed very successfully with normal males for normal females and the latter when mated with sterile males laid sterile eggs. A field trial was arranged in a small isolated village near Bobo-Dioulasso, Upper Volta; the trial took place at the end of the rainy season against a declining population of species A. Sterile males were produced from the cross between species B males and An. melas females; this cross yielded an F1 generation mainly composed of sterile males with a sporadic occurrence of a sometimes significant proportion of females which were reproductively normal. The F1 generation was reared to pupae and these were released into existing breeding places and later from artificial containers. Some 300 000 pupae were so released over a period of 2 months and adult collections were made periodically from inside houses and from outside shelters after releases started. Of the males caught, 75% proved to be sterile while nearly 6% of normal-looking ovipositions failed to hatch. However 2.5% of them proved to be from hybrid females and not from species A females. In control villages, 1.35% of normal-looking ovipositions did not hatch. It was concluded from this field trial that the sterile males were not mating on any significant scale with the natural species A females. This could have been due to a number of factors but the most important is considered to be an ethological one—a mating barrier preventing mating between introduced sterile males and natural females. This is strongly expressed under natural conditions but not operative in the limited confines of a cage. The use of a cross between two species against a third species may well have enhanced this barrier. PMID:5309520

  15. A 3D analysis of flight behavior of Anopheles gambiae sensu stricto malaria mosquitoes in response to human odor and heat.

    PubMed

    Spitzen, Jeroen; Spoor, Cornelis W; Grieco, Fabrizio; ter Braak, Cajo; Beeuwkes, Jacob; van Brugge, Sjaak P; Kranenbarg, Sander; Noldus, Lucas P J J; van Leeuwen, Johan L; Takken, Willem

    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 Anopheles gambiae sensu stricto to human odor and heat. Flight-path characteristics in a wind tunnel (flow 20 cm/s) were quantified in three dimensions. With wind as the only stimulus (control), short and close to straight upwind flights were recorded. With heat alone, flights were similarly short and direct. The presence of human odor, in contrast, caused prolonged and highly convoluted flight patterns. The combination of odor+heat resulted in longer flights with more landings on the source than to either cue alone. Flight speed was greatest (mean groundspeed 27.2 cm/s) for odor+heat. Odor alone resulted in decreased flight speed when mosquitoes arrived within 30 cm of the source whereas mosquitoes exposed to odor+heat maintained a high flight speed while flying in the odor plume, until they arrived within 15 cm of the source. Human odor evoked an increase in crosswind flights with an additive effect of heat at close range (<15 cm) to the source. This was found for both horizontal and vertical flight components. However, mosquitoes nevertheless made upwind progress when flying in the odor+heat generated plume, suggesting that mosquitoes scan their environment intensively while they progress upwind towards their host. These observations may help to improve the efficacy of trapping systems for malaria mosquitoes by (1) optimizing the site of odor release relative to trap entry and (2) adding a heat source which enhances a landing response.

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

    PubMed Central

    Spitzen, Jeroen; Spoor, Cornelis W.; Grieco, Fabrizio; ter Braak, Cajo; Beeuwkes, Jacob; van Brugge, Sjaak P.; Kranenbarg, Sander; Noldus, Lucas P. J. J.; van Leeuwen, Johan L.; Takken, Willem

    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 Anopheles gambiae sensu stricto to human odor and heat. Flight-path characteristics in a wind tunnel (flow 20 cm/s) were quantified in three dimensions. With wind as the only stimulus (control), short and close to straight upwind flights were recorded. With heat alone, flights were similarly short and direct. The presence of human odor, in contrast, caused prolonged and highly convoluted flight patterns. The combination of odor+heat resulted in longer flights with more landings on the source than to either cue alone. Flight speed was greatest (mean groundspeed 27.2 cm/s) for odor+heat. Odor alone resulted in decreased flight speed when mosquitoes arrived within 30 cm of the source whereas mosquitoes exposed to odor+heat maintained a high flight speed while flying in the odor plume, until they arrived within 15 cm of the source. Human odor evoked an increase in crosswind flights with an additive effect of heat at close range (<15 cm) to the source. This was found for both horizontal and vertical flight components. However, mosquitoes nevertheless made upwind progress when flying in the odor+heat generated plume, suggesting that mosquitoes scan their environment intensively while they progress upwind towards their host. These observations may help to improve the efficacy of trapping systems for malaria mosquitoes by (1) optimizing the site of odor release relative to trap entry and (2) adding a heat source which enhances a landing response. PMID:23658792

  17. Cytotoxicity of the Bacillus thuringiensis Cry4B toxin is mediated by the cadherin receptor BT-R₃ of Anopheles gambiae.

    PubMed

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

    2013-07-01

    We demonstrate for the first time the selective cytotoxicity of Bacillus thuringiensis subsp. israelensis Cry4B toxin mediated by BT-R₃ using a cell-based system, which employs High Five insect cells stably expressing BT-R₃. Discovery and validation of BT-R₃ as the Cry4B receptor was accomplished using a web-based computational pipeline platform that facilitates high-throughput insecticidal target identification utilizing the Anopheles gambiae genome. Once the Cry4B toxin binds to the BT-R₃ receptor, a cell death pathway is manifested by sequential cytological changes that include membrane blebbing, cell swelling and lysis. Cry4B toxin associates with cell membrane in both oligomeric and monomeric forms. Monomeric toxin binds specifically to BT-R₃ whereas oligomer interacts with cell membrane non-specifically. Cytotoxicity and cell death are the direct result of binding of toxin monomer to BT-R₃. The oligomeric form of Cry4B toxin is not involved in cell death. Both the location of the toxin-binding region within BT-R₃ and its structural motif are critical to the binding affinity and specificity of the toxin. The toxin-binding region of BT-R₃ appears to be located in EC11, the most membrane proximal EC module within the extracellular domain. It is characterized by the presence of two highly conserved amino acid sequences within their N- and C-termini that flank EC11. These sequences represent signature motifs that mark the toxin-binding function in BT-R₃. The two sequences form two adjacent β-strands within the β-barrel of EC11, the positioning of which is a hallmark of all Cry toxin receptors thus far reported.

  18. Larvicidal activity of neem and karanja oil cakes against mosquito vectors, Culex quinquefasciatus (say), Aedes aegypti (L.) and Anopheles stephensi (L.).

    PubMed

    Shanmugasundaram, R; Jeyalakshmi, T; Dutt, M Sunil; Murthy, P Balakrishna

    2008-01-01

    Larvicidal effect of neem (Azadirachta indica) and karanja (Pongamia glabra) oil cakes (individuals and combination) was studied against mosquito species. Both the oil cakes showed larvicidal activity against the mosquito species tested. The combination of neem and karanja oil cakes in equal proportion proved to have better effect than the individual treatments. The combination of the two oil cakes recorded an LC95 of 0.93, 0.54 and 0.77% against the mosquitoes, Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi respectively The increase in efficacy of the combination treatment over individuals in all the mosquito larvae tested was found to range about 4 to 10 fold in terms of LC50 and 2 to 6 fold in terms of LC95.

  19. Larvicidal, ovicidal, and adulticidal efficacy of Erythrina indica (Lam.) (Family: Fabaceae) against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Govindarajan, Marimuthu; Sivakumar, Rajamohan

    2014-02-01

    Mosquitoes are the major vector for the transmission of malaria, dengue fever, yellow fever, filariasis, schistosomiasis, and Japanese encephalitis. Mosquito control is facing a threat because of the emergence of resistance to synthetic insecticides. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal, ovicidal, and adulticidal potential of the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts from the medicinal plant Erythrina indica against the medically important mosquito vectors, 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 methanol extract of leaf of E. indica against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus with the LC50 and LC90 values of 69.43, 75.13, and 91.41 ppm and 125.49, 134.31, and 167.14 ppm, respectively. 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 E. indica against A. stephensi, A. aegypti, and C. quinquefasciatus exerted 100 % mortality (zero hatchability) at 150, 200, and 250 ppm, respectively. Control eggs showed above 99.3-100 % hatchability. 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

  20. Low-cost and eco-friendly green synthesis of silver nanoparticles using Feronia elephantum (Rutaceae) against Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti (Diptera: Culicidae).

    PubMed

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

    2014-05-01

    Mosquitoes transmit serious human diseases, causing millions of deaths every year. The use of synthetic insecticides to control vector mosquitoes has caused physiological resistance and adverse environmental effects in addition to high operational cost. Insecticides of synthesized natural products for vector control have been a priority in this area. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Feronia elephantum plant leaf extract against late third-instar larvae of Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus was determined. The range of concentrations of synthesized AgNPs (5, 10, 15, 20, and 25 μg mL(-1)) and aqueous leaf extract (25, 50, 75, 100, and 125 μg mL(-1)) were tested against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of F. elephantum for all three important vector mosquitoes. The synthesized AgNPs from F. elephantum were highly toxic than crude leaf aqueous extract to three important vector mosquito species. The results were recorded from UV-visible spectroscopy, Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM), and energy-dispersive X-ray spectroscopy analysis (EDX). Synthesized AgNPs against the vector mosquitoes A. stephensi, A. aegypti, and C. quinquefasciatus had the following LC50 and LC90 values: A. stephensi had LC50 and LC90 values of 11.56 and 20.56 μg mL(-1); A. aegypti had LC50 and LC90 values of 13.13 and 23.12 μg mL(-1); and C. quinquefasciatus had LC50 and LC90 values of 14.19 and 24.30 μg mL(-1). No mortality was observed in the control. These results suggest that the green synthesis of silver nanoparticles using F. elephantum has the potential to be used as an ideal eco-friendly approach for the control of A. stephensi, A. aegypti, and C

  1. Larvicidal and repellent properties of Streptomyces sp. VITJS4 crude extract against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Naine, S Jemimah; Devi, C Subathra

    2014-01-01

    The aim of the present study was to assess the larvicidal and repellent properties of marine Streptomyces sp. VITJS4 crude extracts. The marine soil samples were collected from the Puducherry coast, Tamil Nadu, India. The isolate Streptomyces sp. VITJS4 was taxonomically characterized and identified. The ethyl acetate crude extract tested for larvicidal property showed 100% mortality for all the 3 species after 24 h exposure against the early fourth instar larvae of malarial vector--Anopheles stephensi at 50% and 90% lethal concentration (LC50 = 132.86, LC90 396.14 ppm); dengue vector--Aedes aegypti (LC50 = 112.78, LC90 336.42 ppm) and filariasis vector--Culex quinquefasciatus (LC50 = 156.53, LC90 468.37 ppm). The Streptomyces sp. VITJS4 solvent extracts of hexane, ethyl acetate, benzene, chloroform and methanol were tested for repellent activity against A. stephensi, A. aegypti and C. quinquefasciatus. The ethyl acetate extract showed complete protection for 210 min at 6 mg/cm2 against these mosquito bites. The crude extract was analyzed further for Fourier Transform-infrared spectroscopy (FT-IR) analysis. In addition to the importance of bioactive compounds, the utilization of Streptomyces sp. VITJS4 crude extracts revealed effective larvicidal and repellent activity against the vectors, which perhaps represents a promising tool in the management of mosquito control. PMID:25546945

  2. Ovicidal and adulticidal potential of leaf and seed extract of Albizia lebbeck (L.) Benth. (Family: Fabaceae) against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi (Diptera: Culicidae).

    PubMed

    Govindarajan, Marimuthu; Rajeswary, Mohan

    2015-05-01

    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. In the present study, hexane, benzene, chloroform, ethyl acetate, and methanol extracts of leaf and seed of Albizia lebbeck were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. 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. One hundred percent mortality was observed at 250, 200, and 150 ppm for leaf methanol extract and 375, 300, and 225 ppm for seed methanol extract of A. lebbeck against C. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The adulticidal activity of plant leaf and seed extracts showed moderate toxic effect on the adult mosquitoes after 24 h of exposure period. However, the highest adulticidal activity was observed in the leaf methanol extract of A. lebbeck against An. stephensi where the LC₅₀ and LC₉₀ values were 65.12 and 117.70 ppm, respectively. Compared to leaf extracts, seeds have low potency against three mosquito species. No mortality was recorded in the control. Our data suggest that the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts of A. lebbeck have the potential to be used as an eco-friendly approach for the control of the An. stephensi, Ae. aegypti, and C. quinquefasciatus. These results suggest that the leaf and seed extracts have the potential to be used as an ideal

  3. Ovicidal and adulticidal potential of leaf and seed extract of Albizia lebbeck (L.) Benth. (Family: Fabaceae) against Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi (Diptera: Culicidae).

    PubMed

    Govindarajan, Marimuthu; Rajeswary, Mohan

    2015-05-01

    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. In the present study, hexane, benzene, chloroform, ethyl acetate, and methanol extracts of leaf and seed of Albizia lebbeck were assayed for their toxicity against three important vector mosquitoes, viz., Culex quinquefasciatus, Aedes aegypti, and Anopheles stephensi. 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. One hundred percent mortality was observed at 250, 200, and 150 ppm for leaf methanol extract and 375, 300, and 225 ppm for seed methanol extract of A. lebbeck against C. quinquefasciatus, Ae. aegypti, and An. stephensi, respectively. The adulticidal activity of plant leaf and seed extracts showed moderate toxic effect on the adult mosquitoes after 24 h of exposure period. However, the highest adulticidal activity was observed in the leaf methanol extract of A. lebbeck against An. stephensi where the LC₅₀ and LC₉₀ values were 65.12 and 117.70 ppm, respectively. Compared to leaf extracts, seeds have low potency against three mosquito species. No mortality was recorded in the control. Our data suggest that the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts of A. lebbeck have the potential to be used as an eco-friendly approach for the control of the An. stephensi, Ae. aegypti, and C. quinquefasciatus. These results suggest that the leaf and seed extracts have the potential to be used as an ideal

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

    PubMed

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

    2015-05-01

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

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

    PubMed Central

    2013-01-01

    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 a greater mean and

  6. Effects of an Opal Termination Codon Preceding the nsP4 Gene Sequence in the O'Nyong-Nyong Virus Genome on Anopheles gambiae Infectivity

    PubMed Central

    Myles, Kevin M.; Kelly, Cindy L. H.; Ledermann, Jeremy P.; Powers, Ann M.

    2006-01-01

    The genomic RNA of an alphavirus encodes four different nonstructural proteins, nsP1, nsP2, nsP3, and nsP4. The polyprotein P123 is produced when translation terminates at an opal termination codon between nsP3 and nsP4. The polyprotein P1234 is produced when translational readthrough occurs or when the opal termination codon has been replaced by a sense codon in the alphavirus genome. Evolutionary pressures appear to have maintained genomic sequences encoding both a stop codon (opal) and an open reading frame (arginine) as a general feature of the O'nyong-nyong virus (ONNV) genome, indicating that both are required at some point. Alternate replication of ONNVs in both vertebrate and invertebrate hosts may determine predominance of a particular codon at this locus in the viral quasispecies. However, no systematic study has previously tested this hypothesis in whole animals. We report here the results of the first study to investigate in a natural mosquito host the functional significance of the opal stop codon in an alphavirus genome. We used a full-length cDNA clone of ONNV to construct a series of mutants in which the arginine between nsP3 and nsP4 was replaced with an opal, ochre, or amber stop codon. The presence of an opal stop codon upstream of nsP4 nearly doubled (75.5%) the infectivity of ONNV over that of virus possessing a codon for the amino acid arginine at the corresponding position (39.8%). Although the frequency with which the opal virus disseminated from the mosquito midgut did not differ significantly from that of the arginine virus on days 8 and 10, dissemination did began earlier in mosquitoes infected with the opal virus. Although a clear fitness advantage is provided to ONNV by the presence of an opal codon between nsP3 and nsP4 in Anopheles gambiae, sequence analysis of ONNV RNA extracted from mosquito bodies and heads indicated codon usage at this position corresponded with that of the virus administered in the blood meal. These results

  7. Effects of an opal termination codon preceding the nsP4 gene sequence in the O'Nyong-Nyong virus genome on Anopheles gambiae infectivity.

    PubMed

    Myles, Kevin M; Kelly, Cindy L H; Ledermann, Jeremy P; Powers, Ann M

    2006-05-01

    The genomic RNA of an alphavirus encodes four different nonstructural proteins, nsP1, nsP2, nsP3, and nsP4. The polyprotein P123 is produced when translation terminates at an opal termination codon between nsP3 and nsP4. The polyprotein P1234 is produced when translational readthrough occurs or when the opal termination codon has been replaced by a sense codon in the alphavirus genome. Evolutionary pressures appear to have maintained genomic sequences encoding both a stop codon (opal) and an open reading frame (arginine) as a general feature of the O'nyong-nyong virus (ONNV) genome, indicating that both are required at some point. Alternate replication of ONNVs in both vertebrate and invertebrate hosts may determine predominance of a particular codon at this locus in the viral quasispecies. However, no systematic study has previously tested this hypothesis in whole animals. We report here the results of the first study to investigate in a natural mosquito host the functional significance of the opal stop codon in an alphavirus genome. We used a full-length cDNA clone of ONNV to construct a series of mutants in which the arginine between nsP3 and nsP4 was replaced with an opal, ochre, or amber stop codon. The presence of an opal stop codon upstream of nsP4 nearly doubled (75.5%) the infectivity of ONNV over that of virus possessing a codon for the amino acid arginine at the corresponding position (39.8%). Although the frequency with which the opal virus disseminated from the mosquito midgut did not differ significantly from that of the arginine virus on days 8 and 10, dissemination did began earlier in mosquitoes infected with the opal virus. Although a clear fitness advantage is provided to ONNV by the presence of an opal codon between nsP3 and nsP4 in Anopheles gambiae, sequence analysis of ONNV RNA extracted from mosquito bodies and heads indicated codon usage at this position corresponded with that of the virus administered in the blood meal. These results

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

    PubMed

    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

  9. Examining the impact of larval source management and insecticide-treated nets using a spatial agent-based model of Anopheles gambiae and a landscape generator tool

    PubMed Central

    2013-01-01

    Background Agent-based models (ABMs) have been used to estimate the effects of malaria-control interventions. Early studies have shown the efficacy of larval source management (LSM) and insecticide-treated nets (ITNs) as vector-control interventions, applied both in isolation and in combination. However, the robustness of results can be affected by several important modelling assumptions, including the type of boundary used for landscapes, and the number of replicated simulation runs reported in results. Selection of the ITN coverage definition may also affect the predictive findings. Hence, by replication, independent verification of prior findings of published models bears special importance. Methods A spatially-explicit entomological ABM of Anopheles gambiae is used to simulate the resource-seeking process of mosquitoes in grid-based landscapes. To explore LSM and replicate results of an earlier LSM study, the original landscapes and scenarios are replicated by using a landscape generator tool, and 1,800 replicated simulations are run using absorbing and non-absorbing boundaries. To explore ITNs and evaluate the relative impacts of the different ITN coverage schemes, the settings of an earlier ITN study are replicated, the coverage schemes are defined and simulated, and 9,000 replicated simulations for three ITN parameters (coverage, repellence and mortality) are run. To evaluate LSM and ITNs in combination, landscapes with varying densities of houses and human populations are generated, and 12,000 simulations are run. Results General agreement with an earlier LSM study is observed when an absorbing boundary is used. However, using a non-absorbing boundary produces significantly different results, which may be attributed to the unrealistic killing effect of an absorbing boundary. Abundance cannot be completely suppressed by removing aquatic habitats within 300 m of houses. Also, with density-dependent oviposition, removal of insufficient number of aquatic

  10. Larvicidal activity of silver nanoparticles synthesized using Pergularia daemia plant latex against Aedes aegypti and Anopheles stephensi and nontarget fish Poecillia reticulata.

    PubMed

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

    2012-08-01

    In present study, the bioactivity of latex-producing plant Pergularia daemia as well as synthesized silver nanoparticles (AgNPs) against the larval instars of Aedes aegypti and Anopheles stephensi mosquito larvae was determined. The range of concentrations of plant latex (1,000, 500, 250, 125, 62.25, and 31.25 ppm) and AgNPs (10, 5, 2.5, 1.25, 0.625, and 0.3125 ppm) were prepared. The LC(50) and LC(90) values for first, second, third, and fourth instars of synthesized AgNPs-treated first, second, third, and fourth instars of A. aegypti (LC(50) = 4.39, 5.12, 5.66, 6.18; LC(90) = 9.90, 11.13, 12.40, 12.95 ppm) and A. stephensi (LC(50) = 4.41, 5.35, 5.91, 6.47; LC(90) = 10.10, 12.04, 13.05, 14.08 ppm) were found many fold lower than crude latex-treated A. aegypti (LC(50) = 55.13, 58.81, 75.66, 94.31; LC(90) = 113.00, 118.25, 156.95, 175.71 ppm) and A. stephensi (LC(50) = 81.47, 92.09, 96.07, 101.31; LC(90) = 159.51, 175.97, 180.67, 190.42 ppm). The AgNPs did not exhibit any noticeable effects on Poecillia reticulata after either 24 or 48 h of exposure at their LC(50) and LC(90) values against fourth-instar larvae of A. aegypti and A. stephensi. The UV-visible analysis shows absorbance for AgNPs at 520 nm. TEM reveals spherical shape of synthesized AgNPs. Particle size analysis revealed that the size of particles ranges from 44 to 255 nm with average size of 123.50 nm. AgNPs were clearly negatively charged (zeta potential -27.4 mV). This is the first report on mosquito larvicidal activity P. daemia-synthesized AgNPs.

  11. Larvicidal, pupicidal, repellent and adulticidal activity of Citrus sinensis orange peel extract against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Murugan, Kadarkarai; Mahesh Kumar, Palanisamy; Kovendan, Kalimuthu; Amerasan, Duraisamy; Subrmaniam, Jayapal; Hwang, Jiang-Shiou

    2012-10-01

    Mosquitoes are the carriers of severe and well-known illnesses such as malaria, arboviral encephalitis, dengue fever, chikunguniya fever, West Nile virus and yellow fever. These diseases produce significant morbidity and mortality in humans and livestock around the world. The present study explored the effects of orange peel ethanol extract of Citrus sinensis on larvicidal, pupicidal, repellent and adulticidal activity against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus. The orange peel material was shade dried at room temperature and powdered coarsely. From orange peel, 300 g powdered was macerated with 1 L of ethanol sequentially for a period of 72 h each and filtered. The yields of the orange peel ethanol crude extract of C. sinensis 13.86 g, respectively. The extracts were concentrated at reduced temperature on a rotary vacuum evaporator and stored at a temperature of 4 °C. The larvicidal, pupicidal and adult mortality was observed after 24 h of exposure; no mortality was observed in the control group. For C. sinensis, the median lethal concentration values (LC(50)) observed for the larvicidal and pupicidal activities against mosquito vector species A. stephensi first to fourth larval instars and pupae were 182.24, 227.93, 291.69, 398.00 and 490.84 ppm; A. aegypti values were 92.27, 106.60, 204.87, 264.26, 342.45, 436.93 and 497.41 ppm; and C. quinquefasciatus values were 244.70, 324.04, 385.32, 452.78 and 530.97 ppm, respectively. The results of maximum repellent activity were observed at 450 ppm in ethanol extracts of C. sinensis and the mean complete protection time ranged from 150 to 180 min was tested. The ethanol extract of C. sinensis showed 100% repellency in 150 min and showed complete protection in 90 min at 350 ppm against A. stephensi, A. aegypti and C. quinquefasciatus, respectively. The adult mortality was found in ethanol extract of C. sinensis with the LC(50) and LC(90) values of 272.19 and 457.14 ppm, A. stephensi; 289.62 and

  12. Dose–response tests and semi-field evaluation of lethal and sub-lethal effects of slow release pyriproxyfen granules (Sumilarv®0.5G) for the control of the malaria vectors Anopheles gambiae sensu lato

    PubMed Central

    2013-01-01

    Background Recently research has shown that larviciding can be an effective tool for integrated malaria vector control. Nevertheless, the uptake of this intervention has been hampered by the need to re-apply larvicides frequently. There is a need to explore persistent, environmentally friendly larvicides for malaria vector control to reduce intervention efforts and costs by reducing the frequency of application. In this study, the efficacy of a 0.5% pyriproxyfen granule (Surmilarv®0.5G, Sumitomo Chemicals) was assessed for the control of Anopheles gambiae sensu stricto and Anopheles arabiensis, the major malaria vectors in sub-Saharan Africa. Methods Dose–response and standardized field tests were implemented following standard procedures of the World Health Organization’s Pesticide Evaluation Scheme to determine: (i) the susceptibility of vectors to this formulation; (ii) the residual activity and appropriate retreatment schedule for field application; and, (iii) sub-lethal impacts on the number and viability of eggs laid by adults after exposure to Sumilarv®0.5G during larval development. Results Anopheles gambiae s.s. and An. arabiensis were highly susceptible to Sumilarv®0.5G. Estimated emergence inhibition (EI) values were very low and similar for both species. The minimum dosage that completely inhibited adult emergence was between 0.01-0.03 parts per million (ppm) active ingredient (ai). Compared to the untreated control, an application of 0.018 ppm ai prevented 85% (95% confidence interval (CI) 82%-88%) of adult emergence over six weeks under standardized field conditions. A fivefold increase in dosage of 0.09 ppm ai prevented 97% (95% CI 94%-98%) emergence. Significant sub-lethal effects were observed in the standardized field tests. Female An. gambiae s.s. that were exposed to 0.018 ppm ai as larvae laid 47% less eggs, and females exposed to 0.09 ppm ai laid 74% less eggs than females that were unexposed to the treatment. Furthermore, 77

  13. Status of pyrethroid resistance in Anopheles gambiae s. s. M form prior to the scaling up of Long Lasting Insecticidal Nets (LLINs) in Adzopé, Eastern Côte d’Ivoire

    PubMed Central

    2012-01-01

    Background The growing development of pyrethroid resistance constitutes a serious threat to malaria control programmes and if measures are not taken in time, resistance may compromise control efforts in the foreseeable future. Prior to Long Lasting Insecticidal Nets (LLINs) distribution in Eastern Cote d’Ivoire, we conducted bioassays to inform the National Malaria Control Programme of the resistance status of the main malaria vector, Anopheles gambiae s. s. and the need for close surveillance of resistance. Methods Larvae of An. gambiae s. s. were collected in two areas of Adzopé (Port-Bouët and Tsassodji) and reared to adults. WHO susceptibility tests with impregnated filter papers were carried out to detect resistance to three pyrethroids commonly used to develop LLINs: permethrin 1%, deltamethrin 0.05% and lambda-cyhalothrin 0.05%. Molecular assays were conducted to detect M and S forms and the L1014F kdr allele in individual mosquitoes. Results Resistance, at various degrees was detected in both areas of Adzopé. Overall, populations of An. gambiae at both sites surveyed showed equivalent frequency of the L1014F kdr allele (0.67) but for all tested pyrethroids, there were significantly higher survival rates for mosquitoes from Tsassodji (32–58%) than those from Port-Bouët (3–32%) (p < 0.001), indicating the implication of resistance mechanisms other than kdr alone. During the survey period (May–June) in this forested area of Côte d’Ivoire, An. gambiae s. s. found were exclusively of the M form and were apparently selected for pyrethroid resistance through agricultural and household usage of insecticides. Conclusion Prior to LLINs scaling up in Eastern Côte d’Ivoire, resistance was largely present at various levels in An. gambiae. Underlying mechanisms included the high frequency of the L1014F kdr mutation and other unidentified components, probably metabolic detoxifiers. Their impact on the efficacy of the planned strategy (LLINs) in the

  14. Mosquito larvicidal potential of silver nanoparticles synthesized using Chomelia asiatica (Rubiaceae) against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Muthukumaran, Udaiyan; Govindarajan, Marimuthu; Rajeswary, Mohan

    2015-03-01

    Mosquitoes transmit serious human diseases, causing millions of deaths every year. Mosquito control is to enhance the health and quality of life of county residents and visitors through the reduction of mosquito populations. Mosquito control is a serious concern in developing countries like India due to the lack of general awareness, development of resistance, and socioeconomic reasons. Today, nanotechnology is a promising research domain which has a wide ranging application in vector control programs. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, larvicidal activity of aqueous leaf extract and silver nanoparticles (AgNPs) synthesized using C. asiatica plant leaves against late third instar larvae of Anopheles stephensi, Aedes aegypti, and Cx. quinquefasciatus. The range of varying concentrations of synthesized AgNPs (8, 16, 24, 32, and 40 μg/mL) and aqueous leaf extract (40, 80, 120, 160, and 200 μg/mL) were tested against the larvae of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus. The synthesized AgNPs from C. asiatica were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV-Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy analysis (EDX). Considerable mortality was evident after the treatment of C. asiatica for all three important vector mosquitoes. The LC50 and LC90 values of C. asiatica aqueous leaf extract appeared to be effective against An. stephensi (LC50, 90.17 μg/mL; LC90, 165.18 μg/mL) followed by Ae. aegypti (LC50, 96.59 μg/mL; LC90, 173.83 μg/mL) and Cx. quinquefasciatus (LC50, 103.08 μg/mL; LC90, 183.16 μg/mL). Synthesized AgNPs against the vector mosquitoes of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus had the following LC50 and LC90

  15. Mosquito larvicidal potential of silver nanoparticles synthesized using Chomelia asiatica (Rubiaceae) against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus (Diptera: Culicidae).

    PubMed

    Muthukumaran, Udaiyan; Govindarajan, Marimuthu; Rajeswary, Mohan

    2015-03-01

    Mosquitoes transmit serious human diseases, causing millions of deaths every year. Mosquito control is to enhance the health and quality of life of county residents and visitors through the reduction of mosquito populations. Mosquito control is a serious concern in developing countries like India due to the lack of general awareness, development of resistance, and socioeconomic reasons. Today, nanotechnology is a promising research domain which has a wide ranging application in vector control programs. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, larvicidal activity of aqueous leaf extract and silver nanoparticles (AgNPs) synthesized using C. asiatica plant leaves against late third instar larvae of Anopheles stephensi, Aedes aegypti, and Cx. quinquefasciatus. The range of varying concentrations of synthesized AgNPs (8, 16, 24, 32, and 40 μg/mL) and aqueous leaf extract (40, 80, 120, 160, and 200 μg/mL) were tested against the larvae of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus. The synthesized AgNPs from C. asiatica were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV-Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, and energy-dispersive X-ray spectroscopy analysis (EDX). Considerable mortality was evident after the treatment of C. asiatica for all three important vector mosquitoes. The LC50 and LC90 values of C. asiatica aqueous leaf extract appeared to be effective against An. stephensi (LC50, 90.17 μg/mL; LC90, 165.18 μg/mL) followed by Ae. aegypti (LC50, 96.59 μg/mL; LC90, 173.83 μg/mL) and Cx. quinquefasciatus (LC50, 103.08 μg/mL; LC90, 183.16 μg/mL). Synthesized AgNPs against the vector mosquitoes of An. stephensi, Ae. aegypti, and Cx. quinquefasciatus had the following LC50 and LC90

  16. Physical features and chitin content of eggs from the mosquito vectors Aedes aegypti, Anopheles aquasalis and Culex quinquefasciatus: Connection with distinct levels of resistance to desiccation.

    PubMed

    Farnesi, Luana Cristina; Menna-Barreto, Rubem Figueiredo Sadok; Martins, Ademir Jesus; Valle, Denise; Rezende, Gustavo Lazzaro

    2015-12-01

    Mosquito eggs are laid in water but freshly laid eggs are susceptible to dehydration, if their surroundings dry out at the first hours of development. During embryogenesis of different mosquito vectors the serosal cuticle, an extracellular matrix, is produced; it wraps the whole embryo and becomes part of the eggshell. This cuticle is an essential component of the egg resistance to desiccation (ERD). However, ERD is variable among species, sustaining egg viability for different periods of time. While Aedes aegypti eggs can survive for months in a dry environment (high ERD), those of Anopheles aquasalis and Culex quinquefasciatus in the same condition last, respectively, for one day (medium ERD) or a few hours (low ERD). Resistance to desiccation is determined by the rate of water loss, dehydration tolerance and total amount of water of a given organism. The ERD variability observed among mosquitoes probably derives from diverse traits. We quantified several attributes of whole eggs, potentially correlated with the rate of water loss: length, width, area, volume, area/volume ratio and weight. In addition, some eggshell aspects were also evaluated, such as absolute and relative weight, weight/area relationship (herein called surface density) and chitin content. Presence of chitin specifically in the serosal cuticle as well as aspects of endochorion external surface were also investigated. Three features could be related to differences on ERD levels: chitin content, directly related to ERD, the increase in the egg volume during embryogenesis and the eggshell surface density, which were both inversely related to ERD. Although data suggest that the amount of chitin in the eggshell is relevant for egg impermeability, the participation of other yet unidentified eggshell attributes must be considered in order to account for the differences in the ERD levels observed among Ae. aegypti, An. aquasalis and Cx. quinquefasciatus. PMID:26514070

  17. 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.

    PubMed

    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.

  18. 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.

    PubMed

    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. PMID:26410042

  19. Larvicidal and Biting Deterrent Activity of Essential Oils of Curcuma longa, Ar-turmerone, and Curcuminoids Against Aedes aegypti and Anopheles quadrimaculatus (Culicidae: Diptera).

    PubMed

    Ali, Abbas; Wang, Yan-Hong; Khan, Ikhlas A

    2015-09-01

    Essential oils and extract of Curcuma longa, ar-turmerone, and curcuminoids were evaluated for their larvicidal and deterrent activity against mosquitoes. Ar-turmerone and curcuminoids constituted 36.9, 24.9 and 50.6% of rhizome oil, leaf oil, and rhizome extract, respectively. Ar-turmerone was the major compound of the rhizome oil (36.9%) and leaf oil (24.9%). The ethanolic extract had 15.4% ar-turmerone with 6.6% bisdesmethoxycurcumin, 6.1% desmethoxycurcumin, and 22.6% curcumin. In in vitro studies, essential oils of the leaf (biting deterrence index [BDI] = 0.98), rhizome (BDI = 0.98), and rhizome ethanolic extract (BDI = 0.96) at 10 µg/cm(2) showed biting deterrent activity similar to DEET at 25 nmol/cm(2) against Aedes aegypti L. Among the pure compounds, ar-turmerone (BDI = 1.15) showed the biting deterrent activity higher than DEET at 25 nmol/cm(2) whereas the activity of other compounds was lower than DEET. In Anopheles quadrimaculatus Say, only ar-turmerone showed deterrent activity similar to DEET. In dose-response bioassay, ar-turmerone showed significantly higher biting deterrence than DEET at all the dosages. Ar-turmerone, at 15 nmol/cm(2), showed activity similar to DEET at 25 nmol/cm(2) and activity at 5 nmol/cm(2) was similar to DEET at 20 and 15 nmol/cm(2). Leaf essential oil with LC(50) values of 1.8 and 8.9 ppm against larvae of An. quadrimaculatus and Ae. aegypti, respectively, showed highest toxicity followed by rhizome oil and ethanolic extract. Among the pure compounds, ar-turmerone with LC(50) values of 2.8 and 2.5 ppm against larvae of An. quadrimaculatus and Ae. aegypti, respectively, was most toxic followed by bisdesmethoxycurcumin, curcumin, and desmethoxycurcumin. PMID:26336212

  20. Larvicidal and Biting Deterrent Activity of Essential Oils of Curcuma longa, Ar-turmerone, and Curcuminoids Against Aedes aegypti and Anopheles quadrimaculatus (Culicidae: Diptera).

    PubMed

    Ali, Abbas; Wang, Yan-Hong; Khan, Ikhlas A

    2015-09-01

    Essential oils and extract of Curcuma longa, ar-turmerone, and curcuminoids were evaluated for their larvicidal and deterrent activity against mosquitoes. Ar-turmerone and curcuminoids constituted 36.9, 24.9 and 50.6% of rhizome oil, leaf oil, and rhizome extract, respectively. Ar-turmerone was the major compound of the rhizome oil (36.9%) and leaf oil (24.9%). The ethanolic extract had 15.4% ar-turmerone with 6.6% bisdesmethoxycurcumin, 6.1% desmethoxycurcumin, and 22.6% curcumin. In in vitro studies, essential oils of the leaf (biting deterrence index [BDI] = 0.98), rhizome (BDI = 0.98), and rhizome ethanolic extract (BDI = 0.96) at 10 µg/cm(2) showed biting deterrent activity similar to DEET at 25 nmol/cm(2) against Aedes aegypti L. Among the pure compounds, ar-turmerone (BDI = 1.15) showed the biting deterrent activity higher than DEET at 25 nmol/cm(2) whereas the activity of other compounds was lower than DEET. In Anopheles quadrimaculatus Say, only ar-turmerone showed deterrent activity similar to DEET. In dose-response bioassay, ar-turmerone showed significantly higher biting deterrence than DEET at all the dosages. Ar-turmerone, at 15 nmol/cm(2), showed activity similar to DEET at 25 nmol/cm(2) and activity at 5 nmol/cm(2) was similar to DEET at 20 and 15 nmol/cm(2). Leaf essential oil with LC(50) values of 1.8 and 8.9 ppm against larvae of An. quadrimaculatus and Ae. aegypti, respectively, showed highest toxicity followed by rhizome oil and ethanolic extract. Among the pure compounds, ar-turmerone with LC(50) values of 2.8 and 2.5 ppm against larvae of An. quadrimaculatus and Ae. aegypti, respectively, was most toxic followed by bisdesmethoxycurcumin, curcumin, and desmethoxycurcumin.

  1. Green synthesis of silver nanoparticles using Sida acuta (Malvaceae) leaf extract against Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti (Diptera: Culicidae).

    PubMed

    Veerakumar, Kaliyan; Govindarajan, Marimuthu; Rajeswary, Mohan

    2013-12-01

    Mosquitoes act as a vector for most of the life-threatening diseases like malaria, yellow fever, dengue fever, chikungunya fever, filariasis, encephalitis, West Nile Virus infection, etc. Under the Integrated Mosquito Management, emphasis was given on the application of alternative strategies in mosquito control. The continuous application of synthetic insecticides causes development of resistance in vector species, biological magnification of toxic substances through the food chain, and adverse effects on environmental quality and nontarget organisms including human health. Application of active toxic agents from plant extracts as an alternative mosquito control strategy was available from ancient times. These are nontoxic, easily available at affordable prices, biodegradable, and show broad-spectrum target-specific activities against different species of vector mosquitoes. In the present study, the larvicidal activity of silver nanoparticles (AgNPs) synthesized using Sida acuta plant leaf extract against late third instar larvae of Culex quinquefasciatus, Anopheles stephensi, and Aedes aegypti was determined. Range of concentrations of synthesized AgNPs (10, 20, 30, 40, and 50 μg/mL) and aqueous leaf extract (50, 100, 150, 200, and 250 μg/mL) were tested against the larvae of C. quinquefasciatus, A. stephensi and A. aegypti. The synthesized AgNPs from S. acuta leaf were highly toxic than crude leaf aqueous extract in three important vector mosquito species. The results were recorded from UV-Vis spectrum, Fourier transform infrared spectroscopy, scanning electron microscopy, transmission electron microscopy, and energy-dispersive X-ray spectroscopy analysis. Larvae were exposed to varying concentrations of aqueous crude extract and synthesized AgNPs for 24 h. Considerable mortality was evident after the treatment of S. acuta for all three important vector mosquitoes. The LC50 and LC90 values of S. acuta aqueous leaf extract appeared to be most effective

  2. 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

    PubMed Central

    2010-01-01

    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 to An. gambiae s.l. females

  3. 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

    PubMed Central

    2012-01-01

    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 resistance should be

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

    PubMed Central

    2010-01-01

    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 achieve

  5. Distribution and Frequency of kdr Mutations within Anopheles gambiae s.l. Populations and First Report of the Ace.1G119S Mutation in Anopheles arabiensis from Burkina Faso (West Africa)

    PubMed Central

    Dabiré, Roch K.; Namountougou, Moussa; Diabaté, Abdoulaye; Soma, Dieudonné D.; Bado, Joseph; Toé, Hyacinthe K.; Bass, Chris; Combary, Patrice

    2014-01-01

    An entomological survey was carried out at 15 sites dispersed throughout the three eco-climatic regions of Burkina Faso (West Africa) in order to assess the current distribution and frequency of mutations that confer resistance to insecticides in An. gambiae s.l. populations in the country. Both knockdown (kdr) resistance mutation variants (L1014F and L1014S), that confer resistance to pyrethroid insecticides, were identified concomitant with the ace-1 G119S mutation confirming the presence of multiple resistance mechanisms in the An. gambiae complex in Burkina Faso. Compared to the last survey, the frequency of the L1014F kdr mutation appears to have remained largely stable and relatively high in all species. In contrast, the distribution and frequency of the L1014S mutation has increased significantly in An. gambiae s.l. across much of the country. Furthermore we report, for the first time, the identification of the ace.1 G116S mutation in An. arabiensis populations collected at 8 sites. This mutation, which confers resistance to organophosphate and carbamate insecticides, has been reported previously only in the An. gambiae S and M molecular forms. This finding is significant as organophosphates and carbamates are used in indoor residual sprays (IRS) to control malaria vectors as complementary strategies to the use of pyrethroid impregnated bednets. The occurrence of the three target-site resistance mutations in both An. gambiae molecular forms and now An. arabiensis has significant implications for the control of malaria vector populations in Burkina Faso and for resistance management strategies based on the rotation of insecticides with different modes of action. PMID:25077792

  6. Chlorfenapyr (A Pyrrole Insecticide) Applied Alone or as a Mixture with Alpha-Cypermethrin for Indoor Residual Spraying against Pyrethroid Resistant Anopheles gambiae sl: An Experimental Hut Study in Cove, Benin

    PubMed Central

    Ngufor, Corine; Critchley, Jessica; Fagbohoun, Josias; N’Guessan, Raphael; Todjinou, Damien; Rowland, Mark

    2016-01-01

    Background Indoor spraying of walls and ceilings with residual insecticide remains a primary method of malaria control. Insecticide resistance in malaria vectors is a growing problem. Novel insecticides for indoor residual spraying (IRS) which can improve the control of pyrethroid resistant malaria vectors are urgently needed. Insecticide mixtures have the potential to improve efficacy or even to manage resistance in some situations but this possibility remains underexplored experimentally. Chlorfenapyr is a novel pyrrole insecticide which has shown potential to improve the control of mosquitoes which are resistant to current WHO-approved insecticides. Method The efficacy of IRS with chlorfenapyr applied alone or as a mixture with alpha-cypermeththrin (a pyrethroid) was evaluated in experimental huts in Cove, Southern Benin against wild free flying pyrethroid resistant Anopheles gambiae sl. Comparison was made with IRS with alpha-cypermethrin alone. Fortnightly 30-minute in situ cone bioassays were performed to assess the residual efficacy of the insecticides on the treated hut walls. Results Survival rates of wild An gambiae from the Cove hut site in WHO resistance bioassays performed during the trial were >90% with permethrin and deltamethrin treated papers. Mortality of free-flying mosquitoes entering the experimental huts was 4% in the control hut. Mortality with alpha-cypermethrin IRS did not differ from the control (5%, P>0.656). The highest mortality was achieved with chlorfenapyr alone (63%). The alpha-cypermethrin + chlorfenapyr mixture killed fewer mosquitoes than chlorfenapyr alone (43% vs. 63%, P<0.001). While the cone bioassays showed a more rapid decline in residual mortality with chlorfenapyr IRS to <30% after only 2 weeks, fortnightly mortality rates of wild free-flying An gambiae entering the chlorfenapyr IRS huts were consistently high (50–70%) and prolonged, lasting over 4 months. Conclusion IRS with chlorfenapyr shows potential to

  7. Efficacy of essential oil from Cananga odorata (Lamk.) Hook.f. & Thomson (Annonaceae) against three mosquito species Aedes aegypti (L.), Anopheles dirus (Peyton and Harrison), and Culex quinquefasciatus (Say).

    PubMed

    Soonwera, Mayura

    2015-12-01

    The essential oil of Cananga odorata flowers was evaluated for oviposition-deterrent, ovicidal, insecticidal, and repellent activities toward three mosquito species: Aedes aegypti, Anopheles dirus, and Culex quinquefasciatus. Oviposition deterrence of the oil was evaluated on gravid females using oviposition deterrence bioassay. The results showed that 10 % Ca. odorata exhibited high percent effective repellency against oviposition at 99.4 % to Ae. aegypti, 97.1 % to An. dirus, and 100 % to Cx. quinquefasciatus. Ca. odorata oil was tested for ovicidal activity. Regression equations revealed that the ovicidal rates were positively correlated with the concentrations of the essential oil. As the concentration of essential oil increased from 1, 5, and up to 10 % concentration, the ovicidal rate increased accordingly. Larvicidal activity of the oils was used on immature stages (third and fourth instar lavae and pupae). The maximum larval mortality was found with 10 % Ca. odorata against immature stages, and there were LC50 values ranged from 10.4 to 10.5 % (for Ae. aegypti), <1 % (for An. dirus), and <1 % (for Cx. quinquefasciatus). Adulticidal properties were evaluated with unfed females. Ten percent Ca. odorata oil had high knockdown rates against the three mosquito species at 96 % (for Ae. aegypti), 98.4 % (for An. dirus), and 100 % (for Cx. quinquefasciatus), with EC50 values of 6.2, 4.7, and 5.4 %, respectively. It gave moderate mortality rates after 24 and 48 h of exposure. Ca. odorata oil was assessed for repellency to females by using the modified K&D module. Ten percent Ca. odorata oil gave the strongest value against Ae. aegypti, An. dirus, and Cx. quinquefasciatus, with percentage repellency of 66, 92, and 90 %, respectively. This study demonstrates the potential for the essential oil of Ca. odorata essential oil to be used as a botanical insecticide against three mosquito species.

  8. Efficacy of essential oil from Cananga odorata (Lamk.) Hook.f. & Thomson (Annonaceae) against three mosquito species Aedes aegypti (L.), Anopheles dirus (Peyton and Harrison), and Culex quinquefasciatus (Say).

    PubMed

    Soonwera, Mayura

    2015-12-01

    The essential oil of Cananga odorata flowers was evaluated for oviposition-deterrent, ovicidal, insecticidal, and repellent activities toward three mosquito species: Aedes aegypti, Anopheles dirus, and Culex quinquefasciatus. Oviposition deterrence of the oil was evaluated on gravid females using oviposition deterrence bioassay. The results showed that 10 % Ca. odorata exhibited high percent effective repellency against oviposition at 99.4 % to Ae. aegypti, 97.1 % to An. dirus, and 100 % to Cx. quinquefasciatus. Ca. odorata oil was tested for ovicidal activity. Regression equations revealed that the ovicidal rates were positively correlated with the concentrations of the essential oil. As the concentration of essential oil increased from 1, 5, and up to 10 % concentration, the ovicidal rate increased accordingly. Larvicidal activity of the oils was used on immature stages (third and fourth instar lavae and pupae). The maximum larval mortality was found with 10 % Ca. odorata against immature stages, and there were LC50 values ranged from 10.4 to 10.5 % (for Ae. aegypti), <1 % (for An. dirus), and <1 % (for Cx. quinquefasciatus). Adulticidal properties were evaluated with unfed females. Ten percent Ca. odorata oil had high knockdown rates against the three mosquito species at 96 % (for Ae. aegypti), 98.4 % (for An. dirus), and 100 % (for Cx. quinquefasciatus), with EC50 values of 6.2, 4.7, and 5.4 %, respectively. It gave moderate mortality rates after 24 and 48 h of exposure. Ca. odorata oil was assessed for repellency to females by using the modified K&D module. Ten percent Ca. odorata oil gave the strongest value against Ae. aegypti, An. dirus, and Cx. quinquefasciatus, with percentage repellency of 66, 92, and 90 %, respectively. This study demonstrates the potential for the essential oil of Ca. odorata essential oil to be used as a botanical insecticide against three mosquito species. PMID:26337270

  9. An age–size reaction norm yields insight into environmental interactions affecting life-history traits: a factorial study of larval development in the malaria mosquito Anopheles gambiae sensu stricto

    PubMed Central

    Phelan, Conan; Rotiberg, Bernard D

    2013-01-01

    Environmental factors frequently act nonindependently to determine growth and development of insects. Because age and size at maturity strongly influence population dynamics, interaction effects among environmental variables complicate the task of predicting dynamics of insect populations under novel conditions. We reared larvae of the African malaria mosquito Anopheles gambiae sensu stricto (s.s.) under three factors relevant to changes in climate and land use: food level, water depth, and temperature. Each factor was held at two levels in a fully crossed design, for eight experimental treatments. Larval survival, larval development time, and adult size (wing length) were measured to indicate the importance of interaction effects upon population-level processes. For age and size at emergence, but not survival, significant interaction effects were detected for all three factors, in addition to sex. Some of these interaction effects can be understood as consequences of how the different factors influence energy usage in the context of a nonindependent relationship between age and size. Experimentally assessing interaction effects for all potential future sets of conditions is intractable. However, considering how different factors affect energy usage within the context of an insect's evolved developmental program can provide insight into the causes of complex environmental effects on populations. PMID:23919132

  10. Re-Visiting Insecticide Resistance Status in Anopheles gambiae from Côte d'Ivoire: A Nation-Wide Informative Survey

    PubMed Central

    Koffi, Alphonsine A.; Ahoua Alou, Ludovic P.; Kabran, Jean-Paul K.; N'Guessan, Raphael; Pennetier, Cédric

    2013-01-01

    Insecticide resistance constitutes a major threat that may undermine current gain in malaria control in most endemic countries. National Malaria Control Programmes (NMCPs) need as much information as possible on the resistance status of malaria vectors and underlying mechanisms in order to implement the most relevant and efficient control strategy. Bioassays, biochemical and molecular analysis were performed on An. gambiae collected in six sentinel sites in Côte d'Ivoire. The sites were selected on the basis of their bioclimatic status and agricultural practices. An. gambiae populations across sites showed high levels of resistance to organochloride, pyrethroid and carbamate insecticides. The kdr and ace-1R mutations were detected in almost all sentinel sites with mosquitoes on the coastal and cotton growing areas mostly affected by these mutations. At almost all sites, the levels of detoxifying enzymes (mixed-function oxidases (MFOs), non-specific esterases (NSE) and glutathione-S-transferases (GSTs)) in An. gambiae populations were significantly higher than the levels found in the susceptible strain Kisumu. Pre-exposure of mosquitoes to PBO, an inhibitor of MFOs and NSEs, significantly increased mortality rates to pyrethroids and carbamates in mosquitoes but resistance in most cases was not fully synergised by PBO, inferring a residual role of additional mechanisms, including kdr and ace-1 site insensitivity. The large distribution of resistance in Côte d'Ivoire raises an important question of whether to continue to deploy pyrethroid-based long-lasting insecticidal nets (LLINs) and insecticide residual spraying (IRS) towards which resistance continues to rise with no guarantee that the level of resistance would not compromise their efficacy. Innovative strategies that combine insecticide and synergists in LLINs or spatially LLIN and an effective non-pyrethroid insecticide for IRS could be in the short term the best practice for the NMCP to manage insecticide

  11. Behavioral responses of catnip (Nepeta cataria l.)by two species of mosquitoes, Aedes aegypti (l.) and Anopheles harrisoni harbach and manguin, in Thailand.

    Technology Transfer Automated Retrieval System (TEKTRAN)

    An investigation of the biological effect of catnip oil (Nepeta cataria L.) on the behavioral response of field collected Ae. aegypti and An. harrisoni were conducted using an automated excito-repellency test system. Aedes aegypti showed significant higher escape rates from the contact chamber at 5%...

  12. The Gambia.

    PubMed

    1982-11-01

    Attention in this discussion of the Republic of the Gambia is directed to the following: the people; geography; history; government and political conditions; economy; foreign relations; and relations between the US and Gambia. In 1980 Gambia's population was estimated to be 619,052; the 1980-81 annual growth rate estimate was 2.8%. The infant mortality rate is 217/1000; life expectancy is 32 years for men and 34 years for women. Almost 85% of the population live in rural areas. The principal ethnic groups are Mandinka, Fula, Wolof, Jola, and Serahuli, English is the official language, although most Gambians speak their native language. The Gambia, located on the bulge of West Africa, is low lying, with a maximum altitude of 73 meters above sea level. From what is known of its early history, the Gambia was once part of the Empire of Ghana and the Kingdom of the Songhais. By the 16th century, Portuguese slave traders and gold seekers had settled in the lower river area. During the late 17th and throughout the 18th century, England and France struggled continously for political and commercial supremacy in the regions of the Senegal and Gambia rivers. The Gambia achieved independence on February 18, 1965, as a constitutional monarchy within the British Commonwealth. Shortly thereafter the government proposed conversion from a monarchy to a republic with an elected president replacing the British monarch as chief of state. In 1970 a referendum was approved by the required majority, and the Gambia became a republic on April 24th. The government is divided into independent executive, legislative, and judicial branches. Executive power is vested in a president, popularly elected for a 5 year term. The Gambia is 1 of the few African countries with a genuine multiparty system. There is no army. The economy is almost totally agricultural. The small manfacturing sector accounts for about 5% of the gross domestic product. Few foreign investors have taken advantage of the

  13. Identity and transfer of male reproductive gland proteins of the dengue vector mosquito, Aedes aegypti: potential tools for control of female feeding and reproduction.

    PubMed

    Sirot, Laura K; Poulson, Rebecca L; McKenna, M Caitlin; Girnary, Hussein; Wolfner, Mariana F; Harrington, Laura C

    2008-02-01

    Male reproductive gland proteins (mRGPs) impact the physiology and/or behavior of mated females in a broad range of organisms. We sought to identify mRGPs of the yellow fever mosquito, Aedes aegypti, the primary vector of dengue and yellow fever viruses. Earlier studies with Ae. aegypti demonstrated that "matrone" (a partially purified male reproductive accessory gland substance) or male accessory gland fluid injected into virgin female Ae. aegypti affect female sexual refractoriness, blood feeding and digestion, flight, ovarian development, and oviposition. Using bioinformatic comparisons to Drosophila melanogaster accessory gland proteins and mass spectrometry of proteins from Ae. aegypti male accessory glands and ejaculatory ducts (AG/ED) and female reproductive tracts, we identified 63 new putative Ae. aegypti mRGPs. Twenty-one of these proteins were found in the reproductive tract of mated females but not of virgin females suggesting that they are transferred from males to females during mating. Most of the putative mRGPs fall into the same protein classes as mRGPs in other organisms, although some appear to be evolving rapidly and lack identifiable homologs in Culex pipiens, Anopheles gambiae, and D. melanogaster. Our results identify candidate male-derived molecules that may have an important influence on behavior, survival, and reproduction of female mosquitoes.

  14. Insecticidal and sterilizing effect of Olyset Duo®, a permethrin and pyriproxyfen mixture net against pyrethroid-susceptible and -resistant strains of Anopheles gambiae s.s.: a release-recapture assay in experimental huts

    PubMed Central

    Djènontin, Armel; Ahoua Alou, Ludovic P.; Koffi, Alphonsine; Zogo, Barnabas; Duarte, Elves; N’Guessan, Raphael; Moiroux, Nicolas; Pennetier, Cédric

    2015-01-01

    In the context of the widespread distribution of pyrethroid resistance among malaria vectors, we did a release-recapture trial in experimental huts to investigate the insecticidal and sterilizing effects of a novel long-lasting net (LN), Olyset® Duo, incorporating a mixture of permethrin (PER) and the insect growth regulator (IGR), pyri-proxyfen (PPF). An LN containing PPF alone and a classic Olyset® Net were tested in parallel as positive controls. The effect of progressive number of holes (6, 30, or 150) that may accrue in nets over time was simulated. We used two laboratory Anopheles gambiae s.s. strains: the susceptible Kisumu strain and the pyrethroid-resistant VK-Per strain having solely kdr as resistance mechanism. The effect of these nets on the reproductive success of blood-fed females that survived the different LNs conditions was recorded. Regardless of the mosquito strain, the LNs containing PPF alone with as many as 30 holes drastically reduced the number of eggs laid by females succeeding in feeding, i.e. fecundity by 98% and egg hatching rate (fertility) by 93% relative to untreated control net. Very few of the resistant females blood fed and survived under the Olyset® Duo with similar number of holes (up to 30) but of these few, the inhibition of reproductive success was 100%. There was no evidence that the Olyset® Duo LN with 150 holes impacted fecundity or fertility of the resistant colony. The efficacy of Olyset® Duo is encouraging and clearly illustrates that this new net might be a promising tool for malaria transmission control and resistance management. PMID:26489479

  15. A heterodimeric complex of the LRR proteins LRIM1 and APL1C regulates complement-like immunity in Anopheles gambiae

    SciTech Connect

    Baxter, Richard H.G.; Steinert, Stefanie; Chelliah, Yogarany; Volohonsky, Gloria; Levashina, Elena A.; Deisenhofer, Johann

    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.

  16. Silencing of P-glycoprotein increases mortality in temephos-treated Aedes aegypti larvae.

    PubMed

    Figueira-Mansur, J; Ferreira-Pereira, A; Mansur, J F; Franco, T A; Alvarenga, E S L; Sorgine, M H F; Neves, B C; Melo, A C A; Leal, W S; Masuda, H; Moreira, M F

    2013-12-01

    Re-emergence of vector-borne diseases such as dengue and yellow fever, which are both transmitted by the Aedes aegypti mosquito, has been correlated with insecticide resistance. P-glycoproteins (P-gps) are ATP-dependent efflux pumps that are involved in the transport of substrates across membranes. Some of these proteins have been implicated in multidrug resistance (MDR). In this study, we identified a putative P-glycoprotein in the Ae. aegypti database based on its significantly high identity with Anopheles gambiae, Culex quinquefasciatus, Drosophila melanogaster and human P-gps. The basal ATPase activity of ATP-binding cassette transporters in larvae was significantly increased in the presence of MDR modulators (verapamil and quinidine). An eightfold increase in Ae. aegypti P-gp (AaegP-gp) gene expression was detected in temephos-treated larvae as determined by quantitative PCR. To analyse the potential role of AaegP-gp in insecticide efflux, a temephos larvicide assay was performed in the presence of verapamil. The results showed an increase of 24% in temephos toxicity, which is in agreement with the efflux reversing effect. RNA interference (RNAi)-mediated silencing of the AaegP-gp gene caused a significant increase in temephos toxicity (57%). In conclusion, we have demonstrated for the first time in insects that insecticide-induced P-gp expression can be involved in the modulation of insecticide efflux. PMID:23980723

  17. Intraspecific DNA variation in nuclear genes of the mosquito Aedes aegypti.

    PubMed

    Morlais, I; Severson, D W

    2003-12-01

    Single nucleotide polymorphisms (SNPs) are an abundant source of genetic variation among individual organisms. To assess the usefulness of SNPs for genome analysis in the yellow fever mosquito, Aedes aegypti, we sequenced 25 nuclear genes in each of three strains and analysed nucleotide diversity. The average frequency of nucleotide variation was 12 SNPs per kilobase, indicating that nucleotide variation in Ae. aegypti is similar to that in other organisms, including Drosophila and the malaria vector Anopheles gambiae. Transition polymorphisms outnumbered transversion polymorphisms, at a ratio of about 2:1. We examined codon usage and confirmed that mutational bias favours G and C ending codons. Codon bias was most pronounced in highly expressed genes. Nucleotide diversity estimates indicated that substitution rates are positively correlated in coding and non-coding regions. Nucleotide diversity varied from one gene to another. The unequal distribution of SNPs among Ae. aegypti nuclear genes suggests that single base variations are non-neutral and are subject to selective constraints. Our analysis showed that ubiquitously expressed genes have lower polymorphism rates and are likely under strong purifying selection, whereas tissue specific genes and genes with a putative role in parasite defence exhibit higher levels of polymorphism that may be associated with diversifying selection. PMID:14986924

  18. Silencing of P-glycoprotein increases mortality in temephos-treated Aedes aegypti larvae.

    PubMed

    Figueira-Mansur, J; Ferreira-Pereira, A; Mansur, J F; Franco, T A; Alvarenga, E S L; Sorgine, M H F; Neves, B C; Melo, A C A; Leal, W S; Masuda, H; Moreira, M F

    2013-12-01

    Re-emergence of vector-borne diseases such as dengue and yellow fever, which are both transmitted by the Aedes aegypti mosquito, has been correlated with insecticide resistance. P-glycoproteins (P-gps) are ATP-dependent efflux pumps that are involved in the transport of substrates across membranes. Some of these proteins have been implicated in multidrug resistance (MDR). In this study, we identified a putative P-glycoprotein in the Ae. aegypti database based on its significantly high identity with Anopheles gambiae, Culex quinquefasciatus, Drosophila melanogaster and human P-gps. The basal ATPase activity of ATP-binding cassette transporters in larvae was significantly increased in the presence of MDR modulators (verapamil and quinidine). An eightfold increase in Ae. aegypti P-gp (AaegP-gp) gene expression was detected in temephos-treated larvae as determined by quantitative PCR. To analyse the potential role of AaegP-gp in insecticide efflux, a temephos larvicide assay was performed in the presence of verapamil. The results showed an increase of 24% in temephos toxicity, which is in agreement with the efflux reversing effect. RNA interference (RNAi)-mediated silencing of the AaegP-gp gene caused a significant increase in temephos toxicity (57%). In conclusion, we have demonstrated for the first time in insects that insecticide-induced P-gp expression can be involved in the modulation of insecticide efflux.

  19. Traditional use of mosquito-repellent plants in western Kenya and their evaluation in semi-field experimental huts against Anopheles gambiae: ethnobotanical studies and application by thermal expulsion and direct burning.

    PubMed

    Seyoum, A; Pålsson, K; Kung'a, S; Kabiru, E W; Lwande, W; Killeen, G F; Hassanali, A; Knols, B G J

    2002-01-01

    Ethnobotanical survey in 2 communities in western Kenya revealed that the most commonly known repellent plants were Ocimum americanum L. (64.1%), Lantana camara L. (17.9%), Tagetes minuta L. (11.3%) and Azadirachta indica A. Juss (8.7%) on Rusinga Island, and Hyptis suaveolens Poit. (49.2%), L. camara (30.9%) and O. basilicum L. (30.4%) in Rambira. Direct burning of plants is the most common method of application for O. americanum (68.8%), L. camara (100%) and O. basilicum (58.8%). Placing branches or whole plants inside houses is most common for H. suaveolens (33.3 and 57.8% for the respective locations), A. indica (66.7 and 100%), and T. minuta (54.8 and 56.0%). The repellency of plants suggested by the ethnobotanical survey and other empirical information was evaluated against the malaria vector Anopheles gambiae s.s. Giles in experimental huts within a screenwalled greenhouse. Thermal expulsion and direct burning were tested as alternative application methods for the selected plants O. americanum, O. kilimandscharicum Guerke, O. suave Willd., L. camara, A. indica, H. suaveolens, Lippia uckambensis Spreng and Corymbia citriodora Hook. When thermally expelled, only H. suaveolens failed to repel mosquitoes, whereas the leaves of C. citriodora (74.5%, P < 0.0001), leaves and seeds of O. suave (53.1%, P < 0.0001) and O. kilimandscharicum (52.0%, P < 0.0001) were the most effective. Leaves of C. citriodora also exhibited the highest repellency (51.3%, P < 0.0001) by direct burning, followed by leaves of L. uckambensis (33.4%, P = 0.0004) and leaves and seeds of O. suave (28.0%, P = 0.0255). The combination of O. kilimandscharicum with L. uckambensis repelled 54.8% of mosquitoes (P < 0.0001) by thermal expulsion. No combination of plants increased repellency by either method. The semi-field system described appears a promising alternative to full-field trials for screening large numbers of candidate repellents without risk of malaria exposure.

  20. Détermination des paramètres bioécologiques et entomologiques d’Anopheles gambiae sl dans la transmission du paludisme à Bandundu-ville, République Démocratique de Congo

    PubMed Central

    Matubi, Emery Metelo; Bukaka, Eric; Luemba, Trésor Bakambana; Situakibanza, Hyppolite; Sangaré, Ibrahim; Mesia, Gauthier; Ngoyi, Dieudonné Mumba; Maniania, Nguya Kalemba; Akikwa, Charles Ngandote; Kanza, Jean Pierre Basilua; Tamfum, Jean-Jacques Muyembe; sudi, Jonas Nagahuedi Bongo

    2015-01-01

    Introduction La présente étude a été menée à Bandundu-ville (RDC) en vue d'identifier les paramètres écologiques et entomologiques modulant la transmission du paludisme ainsi que leur tendance saisonnière dans cette agglomération. Méthodes Cette étude a été réalisée dans la période du 1er juin au 31 décembre 2011. Des prospections des gîtes larvaires d'anophèles avec récolte ont été réalisées, les paramètres physiques, physico-chimiques et environnementaux déterminés. La densité larvaire a été estimée selon une échelle de classes de densité, inspirée de la méthode de Carron pour chaque type de gîtes. Quarante-huit maisons ont été sélectionnées et prospectées pour la récolte des moustiques par pulvérisation intradomicilaire. L'identification des moustiques a été faite sur base des critères morphologiques de Gilles et Demeillon. L'Indice sporozoïtique (Is) a été déterminé par le test ELISA CSP de Plasmodium falciparum à l'Institut National de Recherche Biomédicale selon le protocole de Robert Wirtz. Les autres paramètres entomologiques comme la densité, le taux d'agressivité, le taux d'inoculation entomologique (TIE) ainsi que l'indice de stabilité ont été déterminés selon le protocole de l'OMS. La régression linéaire a été réalisée au seuil de signification de 0,05 pour identifier les déterminants de la densité larvaire. Résultats Cent-sept gîtes larvaires ont été identifiés et caractérisés en 5 types (digues et puits d'eau, collections d'eau maraîchère et concasseurs moellons, marais Régie de distribution d'eau, marais le long des rivières et ruisseaux et flaques d'eau de pluies). La densité larvaire moyenne a été de 117,4±64,1. Quatre mille cinq cents quatre-vingt-huit moustiques ont été capturés et identifiés, parmi lesquels 1.258 Anopheles gambiae sl avec une densité de 8,86, un taux d'agressivité de 1,55 piqûre par homme par nuit, l'Is de 5,6%, un TIE de 0,085 piq

  1. Larvicidal and ovicidal properties of leaf and seed extracts of Delonix elata (L.) Gamble (family: Fabaceae) against malaria (Anopheles stephensi Liston) and dengue (Aedes aegypti Linn.) (Diptera: Culicidae) vector mosquitoes.

    PubMed

    Marimuthu, Govindarajan; Rajamohan, Sivakumar; Mohan, Rajeswari; Krishnamoorthy, Yogalakshmi

    2012-07-01

    Mosquito-borne diseases with an economic impact create loss in commercial and labor outputs, particularly in countries with tropical and subtropical climates. Mosquito control is facing a threat because of the emergence of resistance to synthetic insecticides. Extracts from plants may be alternative sources of mosquito control agents because they constitute a rich source of bioactive compounds that are biodegradable into nontoxic products and potentially suitable for use to control mosquitoes. Insecticides of botanical origin may serve as suitable alternative biocontrol techniques in the future. In view of the recently increased interest in developing plant origin insecticides as an alternative to chemical insecticide, this study was undertaken to assess the larvicidal and ovicidal potential of the crude hexane, benzene, chloroform, ethyl acetate, and methanol solvent extracts from the medicinal plant Delonix elata against the medically important mosquito vectors, Anopheles stephensi and Aedes aegypti (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 methanol extract of leaf of D. elata against the larvae of A. stephensi and A. aegypti with the LC(50) and LC(90) values being 93.59 and 111.83, and 163.69 and 202.77 ppm, respectively. Compared to leaf extracts, seeds have low potency against two mosquitoes with the LC(50) and LC(90) values being 115.28 and 139.04, and 225.07 and 273.03 ppm, respectively. 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. One hundred percent mortality was observed at 300 ppm for leaf methanol extract and 500 ppm for seed

  2. Genomic Analysis of Detoxification Supergene Families in the Mosquito Anopheles sinensis

    PubMed Central

    Zhou, Dan; Liu, Xianmiao; Sun, Yan; Ma, Lei; Shen, Bo; Zhu, Changliang

    2015-01-01

    Anopheles sinensis is an important malaria vector in China and other Southeast Asian countries, and the emergence of insecticide resistance in this mosquito poses a serious threat to the efficacy of malaria control programs. The recently published An. sinensis genome and transcriptome provide an opportunity to understand the molecular mechanisms of insecticide resistance. Analysis of the An. sinensis genome revealed 174 detoxification genes, including 93 cytochrome P450s (P450s), 31 glutathione-S-transferases (GSTs), and 50 choline/carboxylesterases (CCEs). The gene number was similar to that in An. gambiae, but represented a decrease of 29% and 42% compared with Aedes aegypti and Culex quinquefasciatus, respectively. The considerable contraction in gene number in Anopheles mosquitoes mainly occurred in two detoxification supergene families, P450s and CCEs. The available An. sinensis transcriptome was also re-analyzed to further identify key resistance-associated detoxification genes. Among 174 detoxification genes, 124 (71%) were detected. Several candidate genes overexpressed in a deltamethrin-resistant strain (DR-strain) were identified as belonging to the CYP4 or CYP6 family of P450s and the Delta GST class. These generated data provide a basis for identifying the resistance-associated genes of An. sinensis at the molecular level. PMID:26588704

  3. Nigeria Anopheles Vector Database: An Overview of 100 Years' Research

    PubMed Central

    Okorie, Patricia Nkem; McKenzie, F. Ellis; Ademowo, Olusegun George; Bockarie, Moses; Kelly-Hope, Louise

    2011-01-01

    Anopheles mosquitoes are important vectors of malaria and lymphatic filariasis (LF), which are major public health diseases in Nigeria. Malaria is caused by infection with a protozoan parasite of the genus Plasmodium and LF by the parasitic worm Wuchereria bancrofti. Updating our knowledge of the Anopheles species is vital in planning and implementing evidence based vector control programs. To present a comprehensive report on the spatial distribution and composition of these vectors, all published data available were collated into a database. Details recorded for each source were the locality, latitude/longitude, time/period of study, species, abundance, sampling/collection methods, morphological and molecular species identification methods, insecticide resistance status, including evidence of the kdr allele, and P. falciparum sporozoite rate and W. bancrofti microfilaria prevalence. This collation resulted in a total of 110 publications, encompassing 484,747 Anopheles mosquitoes in 632 spatially unique descriptions at 142 georeferenced locations being identified across Nigeria from 1900 to 2010. Overall, the highest number of vector species reported included An. gambiae complex (65.2%), An. funestus complex (17.3%), An. gambiae s.s. (6.5%). An. arabiensis (5.0%) and An. funestus s.s. (2.5%), with the molecular forms An. gambiae M and S identified at 120 locations. A variety of sampling/collection and species identification methods were used with an increase in molecular techniques in recent decades. Insecticide resistance to pyrethroids and organochlorines was found in the main Anopheles species across 45 locations. Presence of P. falciparum and W. bancrofti varied between species with the highest sporozoite rates found in An. gambiae s.s, An. funestus s.s. and An. moucheti, and the highest microfilaria prevalence in An. gambiae s.l., An. arabiensis, and An. gambiae s.s. This comprehensive geo-referenced database provides an essential baseline on Anopheles

  4. Identification of Aedes aegypti Long Intergenic Non-coding RNAs and Their Association with Wolbachia and Dengue Virus Infection

    PubMed Central

    Etebari, Kayvan; Asad, Sultan; Zhang, Guangmei; Asgari, Sassan

    2016-01-01

    Long intergenic non-coding RNAs (lincRNAs) are appearing as an important class of regulatory RNAs with a variety of biological functions. The aim of this study was to identify the lincRNA profile in the dengue vector Aedes aegypti and evaluate their potential role in host-pathogen interaction. The majority of previous RNA-Seq transcriptome studies in Ae. aegypti have focused on the expression pattern of annotated protein coding genes under different biological conditions. Here, we used 35 publically available RNA-Seq datasets with relatively high depth to screen the Ae. aegypti genome for lincRNA discovery. This led to the identification of 3,482 putative lincRNAs. These lincRNA genes displayed a slightly lower GC content and shorter transcript lengths compared to protein-encoding genes. Ae. aegypti lincRNAs also demonstrate low evolutionary sequence conservation even among closely related species such as Culex quinquefasciatus and Anopheles gambiae. We examined their expression in dengue virus serotype 2 (DENV-2) and Wolbachia infected and non-infected adult mosquitoes and Aa20 cells. The results revealed that DENV-2 infection increased the abundance of a number of host lincRNAs, from which some suppress viral replication in mosquito cells. RNAi-mediated silencing of lincRNA_1317 led to enhancement in viral replication, which possibly indicates its potential involvement in the host anti-viral defense. A number of lincRNAs were also differentially expressed in Wolbachia-infected mosquitoes. The results will facilitate future studies to unravel the function of lncRNAs in insects and may prove to be beneficial in developing new ways to control vectors or inhibit replication of viruses in them. PMID:27760142

  5. Laboratory and field evaluation of the impact of washings on the effectiveness of LifeNet®, Olyset® and PermaNet® 2.0 in two areas, where there is a high level of resistance of Anopheles gambiae to pyrethroids, Benin, West Africa

    PubMed Central

    2014-01-01

    Background An investigation carried out in Benin has shown that, in some areas close to rivers where density of mosquitoes is high, long-lasting, insecticidal bed nets (LLINs) are permanently used. In such areas, LLINs are washed every month. Based on this situation, the 20-wash minimum efficacy advised by the manufacturers would be inadequate. The main goal of this study was to evaluate the effectiveness of LifeNet®, Olyset® and Permanet® 2.0 washed several times against Anopheles gambiae sensu stricto (s.s.) populations, which have developed high resistance to pyrethroids. Methods Efficacy of LifeNet®, Olyset® and PermaNet® 2.0 washed 30 and 40 times was expressed in terms of blood-feeding inhibition rate, deterrence, induced exophily and mortality rates. This WHOPES phase II evaluation, conducted in experimental huts in Akron (southern Benin) and in Malanville (northern Benin), was accompanied by WHOPES Phase I evaluation. Results Over 40 successive washes, LifeNet® induced a mortality rate over 80% in phase I. However, beyond 10 washes, Permanet® 2.0 and Olyset induced dramatically reduced mortality rates, respectively 12.5 and 2.5%. With regard to Phase II results, unwashed LifeNet®, LifeNet® and Olyset® washed 30 and 40 times induced a similar exophily rate per study site (at least 58% in Malanville and at least 71% in Akron). Regarding blood feeding inhibition, LifeNet® and Olyset® washed 30 and 40 times significantly reduced wild An. gambiae s.s. blood feeding showing a similar personal protection as unwashed LifeNet®. LifeNet® washed 30 and 40 times induced mortality rates significantly higher than those induced by Olyset® and Permanet® 2.0 (P < 0,05). Conclusion LifeNet®, followed by Olyset®, have shown good efficacy against host-seeking resistant An. gambiae s.s. population in experimental huts in Benin. Lifenet® have shown to be an effective and promising vector control tool to prevent malaria in areas where repeated washings is

  6. Efficacy of Olyset® Duo, a permethrin and pyriproxyfen mixture net against wild pyrethroid-resistant Anopheles gambiae s.s. from Côte d’Ivoire: an experimental hut trial

    PubMed Central

    Koffi, Alphonsine A.; Ahoua Alou, Ludovic P.; Djenontin, Armel; Kabran, Jean-Paul K.; Dosso, Youssouf; Kone, Aboubacar; Moiroux, Nicolas; Pennetier, Cedric

    2015-01-01

    Pyrethroid resistance in malaria vectors has spread across sub-Saharan Africa. Alternative tools and molecules are urgently needed for effective vector control. One of the most promising strategies to prevent or delay the development of resistance is to use at least two molecules having unrelated modes of action in combination in the same bed net. We evaluated in experimental huts in Côte d’Ivoire, a new polyethylene long-lasting insecticidal net (LN) product, Olyset® Duo, incorporating permethrin (PER) and pyriproxyfen (PPF), an insect growth regulator (IGR). PPF alone or in combination with permethrin had a significant impact on fertility (7–12% reduction relative to control) and no effect on fecundity of wild multi-resistant An. gambiae s.s. These results triggered crucial research questions on the behaviour of targeted mosquitoes around the LN. To maximize the sterilizing effect of PPF in the combination, there would be a need for a trade-off between the necessary contact time of the insect with PPF and the surface content of the pyrethroid insecticide that is bioavailable and induces excito-repellency. PMID:26489480

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

    PubMed Central

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

    2015-01-01

    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. PMID:26583525

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

    PubMed

    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. PMID:26583525

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

    PubMed

    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.

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

    PubMed Central

    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

  11. “Saltwater Anopheles gambiae” on Mauritius*

    PubMed Central

    Paterson, H. E.

    1964-01-01

    In this paper the author reports the results of three months' study of the saltwater-breeding member of the Anopheles gambiae complex of sibling species on Mauritius. There is evidence for the views that this form's distribution on the island is limited by the availability of suitable breeding areas, that it does not usually disperse far from the breeding grounds or coast, and that it is probably not an important vector except, perhaps, in the near vicinity of its breeding places. Some new evidence is presented in support of the view that this form (and forms A and B) are distinct species. This turns on the observed close coexistence of these three forms on Mauritius, supported by a theoretical consideration of what would be expected to happen in such circumstances if a system of random mating prevailed. Evidence is given that the Mauritian saltwater-breeding form of the A. gambiae complex is conspecific with the form occurring on the east coast of Africa. The practical importance of reaching general agreement on the evolutionary status of the members of the A. gambiae complex is emphasized. PMID:14278002

  12. Dose and developmental responses of Anopheles merus larvae to salinity.

    PubMed

    White, Bradley J; Kundert, Peter N; Turissini, David A; Van Ekeris, Leslie; Linser, Paul J; Besansky, Nora J

    2013-09-15

    Saltwater tolerance is a trait that carries both ecological and epidemiological significance for Anopheles mosquitoes that transmit human malaria, as it plays a key role in determining their habitat use and ecological distribution, and thus their local contribution to malaria transmission. Here, we lay the groundwork for genetic dissection of this trait by quantifying saltwater tolerance in three closely related cryptic species and malaria vectors from the Afrotropical Anopheles gambiae complex that are known to differ starkly in their tolerance to salinity: the obligate freshwater species A. gambiae and A. coluzzii, and the saltwater-tolerant species A. merus. We performed detailed comparisons of survivorship under varying salinities, using multiple strains of A. gambiae, A. coluzzii and A. merus, as well as F1 progeny from reciprocal crosses of A. merus and A. coluzzii. Additionally, using immunohistochemistry, we compared the location of three ion regulatory proteins (Na(+)/K(+)-ATPase, carbonic anhydrase and Na(+)/H(+)-antiporter) in the recta of A. coluzzii and A. merus reared in freshwater or saline water. As expected, we found that A. merus survives exposure to high salinities better than A. gambiae and A. coluzzii. Further, we found that exposure to a salinity level of 15.85 g NaCl l(-1) is a discriminating dose that kills all A. gambiae, A. coluzzii and A. coluzzii-A. merus F1 larvae, but does not negatively impact the survival of A. merus. Importantly, phenotypic expression of saltwater tolerance by A. merus is highly dependent upon the developmental time of exposure, and based on immunohistochemistry, salt tolerance appears to involve a major shift in Na(+)/K+-ATPase localization in the rectum, as observed previously for the distantly related saline-tolerant species A. albimanus. PMID:23966587

  13. The effect of physical water quality and water level changes on the occurrence and density of Anopheles mosquito larvae around the shoreline of the Koka reservoir, central Ethiopia

    NASA Astrophysics Data System (ADS)

    Teklu, B. M.; Tekie, H.; McCartney, M.; Kibret, S.

    2010-12-01

    Entomological studies to determine the effect of the physical characteristics of mosquito larval breeding water bodies and reservoir water level changes on the occurrence of Anopheles mosquito larvae were conducted in two villages at Koka reservoir in central Ethiopia between August and December 2007. Of the two study villages, Ejersa is located close 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, derived from 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 An. gambiae s.l. in this study was higher in slightly turbid and shallow aquatic habitat than in turbid and relatively deep aquatic habitat. The density of An. 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. There was also a positive correlation between the occurrence of Anopheles larvae with the water and daily minimum atmospheric temperature. Similarly at Ejersa, over the sampling period, there was a positive correlation between falling reservoir water levels and the number of positive breeding habitats. These results confirm that physical characteristics of the water bodies play an important role in the species composition, total Anopheles larval count, and the density of Anopheles mosquitoes. Suitable breeding habitat in the vicinity of the reservoir village was strongly associated with the reservoir. This is particularly important for An

  14. Spatial and temporal heterogeneity of Anopheles mosquitoes and Plasmodium falciparum transmission along the Kenyan coast.

    PubMed

    Mbogo, Charles M; Mwangangi, Joseph M; Nzovu, Joseph; Gu, Weidong; Yan, Guiyan; Gunter, James T; Swalm, Chris; Keating, Joseph; Regens, James L; Shililu, Josephat I; Githure, John I; Beier, John C

    2003-06-01

    The seasonal dynamics and spatial distributions of Anopheles mosquitoes and Plasmodium falciparum parasites were studied for one year at 30 villages in Malindi, Kilifi, and Kwale Districts along the coast of Kenya. Anopheline mosquitoes were sampled inside houses at each site once every two months and malaria parasite prevalence in local school children was determined at the end of the entomologic survey. A total of 5,476 Anopheles gambiae s.l. and 3,461 An. funestus were collected. Species in the An. gambiae complex, identified by a polymerase chain reaction, included 81.9% An. gambiae s.s., 12.8% An. arabiensis, and 5.3% An. merus. Anopheles gambiae s.s. contributed most to the transmission of P. falciparum along the coast as a whole, while An. funestus accounted for more than 50% of all transmission in Kwale District. Large spatial heterogeneity of transmission intensity (< 1 up to 120 infective bites per person per year) resulted in correspondingly large and significantly related variations in parasite prevalence (range = 38-83%). Thirty-two percent of the sites (7 of 22 sites) with malaria prevalences ranging from 38% to 70% had annual entomologic inoculation rates (EIR) less than five infective bites per person per year. Anopheles gambiae s.l. and An. funestus densities in Kwale were not significantly influenced by rainfall. However, both were positively correlated with rainfall one and three months previously in Malindi and Kilifi Districts, respectively. These unexpected variations in the relationship between mosquito populations and rainfall suggest environmental heterogeneity in the predominant aquatic habitats in each district. One important conclusion is that the highly non-linear relationship between EIRs and prevalence indicates that the consistent pattern of high prevalence might be governed by substantial variation in transmission intensity measured by entomologic surveys. The field-based estimate of entomologic parameters on a district level does

  15. Reproductive Isolation among Sympatric Molecular Forms of An. gambiae from Inland Areas of South-Eastern Senegal

    PubMed Central

    Niang, El Hadji Amadou; Konaté, Lassana; Diallo, Mawlouth; Faye, Ousmane; Dia, Ibrahima

    2014-01-01

    The Anopheles gambiae species complex includes at least seven morphologically indistinguishable species, one of which, Anopheles gambiae sensu stricto, is the primary mosquito vector responsible for the transmission of malaria across sub-Saharan Africa. Sympatric ecological diversification of An. gambiae s.s. is in progress within this complex, leading to the emergence of at least two incipient species (the M and S molecular forms now recognized as good species and named An. coluzzii and An. gambiae respectively) that show heterogeneous levels of divergence in most parts of Africa. However, this process seems to have broken down in coastal areas of West Africa at the extreme edge of the distribution. We undertook a longitudinal study to describe An. gambiae s.s. populations collected from two inland transects with different ecological characteristics in south-eastern Senegal. Analysis of samples collected from 20 sites across these two transects showed the M and S molecular forms coexisted at almost all sampled sites. Overall, similar hybridization rates (2.16% and 1.86%) were recorded in the two transects; sites with relatively high frequencies of M/S hybrids (up to 7%) were clustered toward the north-western part of both transects, often near urban settings. Estimated inbreeding indices for this putative speciation event varied spatially (range: 0.52–1), with hybridization rates being generally lower than expected under panmictic conditions. Such observations suggest substantial reproductive isolation between the M and S molecular forms, and further support the ongoing process of speciation in these inland areas. According to a recent reclassification of the An. gambiae complex, the M and S molecular forms from this zone correspond to An. coluzzii and An. gambiae, respectively. There is considerable evidence that these molecular forms differ in their behavioural and ecological characteristics. Detailed study of these characteristics will allow the development

  16. Microbial larvicides for malaria control in The Gambia

    PubMed Central

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

    2007-01-01

    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 larvae for three

  17. Complex genome evolution in Anopheles coluzzii associated with increased insecticide usage in Mali.

    PubMed

    Main, Bradley J; Lee, Yoosook; Collier, Travis C; Norris, Laura C; Brisco, Katherine; Fofana, Abdrahamane; Cornel, Anthony J; Lanzaro, Gregory C

    2015-10-01

    In certain cases, a species may have access to important genetic variation present in a related species via adaptive introgression. These novel alleles may interact with their new genetic background, resulting in unexpected phenotypes. In this study, we describe a selective sweep on standing variation on the X chromosome in the mosquito Anopheles coluzzii, a principal malaria vector in West Africa. This event may have been influenced by the recent adaptive introgression of the insecticide resistance gene known as kdr from the sister species Anopheles gambiae. Individuals carrying both kdr and a nearly fixed X-linked haplotype, encompassing at least four genes including the P450 gene CYP9K1 and the cuticular protein CPR125, have rapidly increased in relative frequency. In parallel, a reproductively isolated insecticide-susceptible A. gambiae population (Bamako form) has been driven to local extinction, likely due to strong selection from increased insecticide-treated bed net usage. PMID:26359110

  18. Native microbiome impedes vertical transmission of Wolbachia in Anopheles mosquitoes

    PubMed Central

    Hughes, Grant L.; Dodson, Brittany L.; Johnson, Rebecca M.; Murdock, Courtney C.; Tsujimoto, Hitoshi; Suzuki, Yasutsugu; Patt, Alyssa A.; Cui, Long; Nossa, Carlos W.; Barry, Rhiannon M.; Sakamoto, Joyce M.; Hornett, Emily A.; Rasgon, Jason L.

    2014-01-01

    Over evolutionary time, Wolbachia has been repeatedly transferred between host species contributing to the widespread distribution of the symbiont in arthropods. For novel infections to be maintained, Wolbachia must infect the female germ line after being acquired by horizontal transfer. Although mechanistic examples of horizontal transfer exist, there is a poor understanding of factors that lead to successful vertical maintenance of the acquired infection. Using Anopheles mosquitoes (which are naturally uninfected by Wolbachia) we demonstrate that the native mosquito microbiota is a major barrier to vertical transmission of a horizontally acquired Wolbachia infection. After injection into adult Anopheles gambiae, some strains of Wolbachia invade the germ line, but are poorly transmitted to the next generation. In Anopheles stephensi, Wolbachia infection elicited massive blood meal-induced mortality, preventing development of progeny. Manipulation of the mosquito microbiota by antibiotic treatment resulted in perfect maternal transmission at significantly elevated titers of the wAlbB Wolbachia strain in A. gambiae, and alleviated blood meal-induced mortality in A. stephensi enabling production of Wolbachia-infected offspring. Microbiome analysis using high-throughput sequencing identified that the bacterium Asaia was significantly reduced by antibiotic treatment in both mosquito species. Supplementation of an antibiotic-resistant mutant of Asaia to antibiotic-treated mosquitoes completely inhibited Wolbachia transmission and partly contributed to blood meal-induced mortality. These data suggest that the components of the native mosquito microbiota can impede Wolbachia transmission in Anopheles. Incompatibility between the microbiota and Wolbachia may in part explain why some hosts are uninfected by this endosymbiont in nature. PMID:25114252

  19. [The Anopheles fauna and the transmission of human malaria in Kinshasa (Zaire)].

    PubMed

    Karch, S; Asidi, N; Manzambi, Z M; Salaun, J J

    1992-01-01

    A longitudinal epidemiological study of malaria and its vectors was conducted in Kinshasa. 264 night-bite collections on human bait (1,056 man nights) and 384 collections of the house-resting fauna were carried out from April 1989 to October 1990. The anophelian fauna was identified and inventoried, 7 Anopheles species were found: Anopheles gambiae, An. funestus, An. paludis, An. hancocki, An. counstani, An. brunnipes, and An. nili. A single species, An. gambiae s. l. is responsible for the transmission of malaria, it represents 93.27% of the anopheline fauna. The average number of anophele bites man day was 16.28 bites/man/night, it varied between 1 b/m/n in urban area to 26.05 b/m/n in semi-rural area. The average of the sporozoite index for An. gambiae was 3.3%, but it varied from 0% in the urban area to 6.52% in the semi-rural area. The entomological inoculation rate (h) was 197 infective bites per year. This rate fluctuated from 1 infective bite each 128 nights in urban area to 1.7 infective night-bite in semi-rural area. Other epidemiological index were also determined: the level of daily survival rate (p = 8.75 days), the vectorial capacity of 17.97 and the Macdonald's stability 3.5 bites on man taken by a vector during its entire lifetime.

  20. Pteridine fluorescence for age determination of Anopheles mosquitoes.

    PubMed

    Wu, D; Lehane, M J

    1999-02-01

    The age structure of mosquito populations is of great relevance to understanding the dynamics of disease transmission and in monitoring the success of control operations. Unfortunately, the ovarian dissection methods currently available for determining the age of adult mosquitoes are technically difficult, slow and may be of limited value, because the proportion of diagnostic ovarioles in the ovary declines with age. By means of reversed-phase HPLC this study investigated the malaria vectors Anopheles gambiae and An. stephensi to see if changes in fluorescent pteridine pigments, which have been used in other insects to determine the age of field-caught individuals, may be useful for age determination in mosquitoes. Whole body fluorescence was inversely proportional to age (P < 0.001, r2 > 91%) up to 30 days postemergence, with the regression values: y = 40580-706x for An. gambiae, and y = 52896-681x for An. stephensi. In both species the main pteridines were 6-biopterin, pterin-6-carboxylic acid and an unidentified fluorescent compound. An. gambiae had only 50-70% as much fluorescence as An. stephensi, and fluorescent compounds were relatively more concentrated in the head than in the thorax (ratios 1:0.8 An. gambiae; 1:0.5 An. stephensi). The results of this laboratory study are encouraging. It seems feasible that this simpler and faster technique of fluorescence quantification could yield results of equivalent accuracy to the interpretation of ovarian dissection. A double-blind field trial comparing the accuracy of this technique to marked, released and recaptured mosquitoes is required to test the usefulness of the pteridine method in the field. PMID:10194749

  1. Rubidium marking of Anopheles mosquitoes detectable by field-capable X-ray spectrometry.

    PubMed

    Wilkins, E E; Smith, S C; Roberts, J M; Benedict, M

    2007-06-01

    We present a mosquito marking technique suitable for mark-release-recapture that can be used with a hand-held, portable X-ray fluorescence (XRF) spectrometer, which is practical for field measurements. Third instar Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) and Anopheles stephensi Liston larvae were cultured to pupation in water containing rubidium (Rb) Cl at concentrations up to 1000 p.p.m. Rb. Anopheles gambiae larvae survived to adulthood at concentrations as high as 1000 p.p.m. Rb but suffered pupal mortality and reduced adult longevity at high concentrations. We were able to culture An. stephensi at Rb concentrations as high as 300 p.p.m. The presence of Rb in adults was evaluated using a portable XRF analyser, and we were able to reliably detect Rb above background levels in 10-day-old females and 4-day-old males at concentrations causing minimal pupal or adult mortality. We observed that Rb marking was not permanent, and the concentration declined significantly as adults aged. The low cost of labelling with RbCl and the field portability of the spectrometer provide a useful means for labelling mosquitoes via breeding sites or in the laboratory for mark-release-recapture experiments.

  2. Vectorial status and insecticide resistance of Anopheles funestus from a sugar estate in southern Mozambique

    PubMed Central

    2011-01-01

    Background The dual problems of rising insecticide resistance in the malaria vectors and increasing human malaria cases since 2001 in southern Mozambique are cause for serious concern. The selection of insecticides for use in indoor residual spraying (IRS) programmes is highly dependent on the extent to which local mosquitoes are susceptible to the approved classes of insecticides. The insecticide resistance status and role in malaria transmission of Anopheles funestus was evaluated at the Maragra Sugar Estate in southern Mozambique where an IRS vector control programme has been in operation for seven years using the carbamate insecticide bendiocarb. Results No Anopheles species were captured inside the sugar estate control area. Anopheles funestus group captured outside of the estate represented 90% (n = 475) of the total collections. Of the specimens identified to species by PCR (n = 167), 95% were An. funestus s.s. One An. rivulorum was identified and seven specimens did not amplify. The Anopheles gambiae complex was less abundant (n = 53) and of those identified (n = 33) 76% were An. arabiensis and 24% An. merus. Insecticide susceptibility tests showed that wild-caught and F-1 family An. funestus were resistant to deltamethrin (32.5% mortality) and lambda-cyhalothrin (14.6% mortality), less so to bendiocarb (71.5% mortality) and fully susceptible to both malathion and DDT (100%). Bendiocarb and pyrethroid resistance was nullified using 4% piperonyl butoxide (Pbo), strongly suggesting that both are mediated by P450 monooxygenase detoxification. ELISA tests of An. funestus for Plasmodium falciparum, gave a sporozoite rate of 6.02% (n = 166). One unidentified member of the An. gambiae complex tested positive for P. falciparum sporozoites. Conclusion Anopheles funestus was found to be the most abundant and principle vector of malaria in this area, with members of the An. gambiae complex being secondary vectors. Despite the continual use of bendiocarb within the

  3. High spatial resolution mapping of malaria transmission risk in the Gambia, west Africa, using LANDSAT TM satellite imagery.

    PubMed

    Bøgh, Claus; Lindsay, Steven W; Clarke, Siân E; Dean, Andy; Jawara, Musa; Pinder, Margaret; Thomas, Christopher J

    2007-05-01

    Understanding local variability in malaria transmission risk is critically important when designing intervention or vaccine trials. Using a combination of field data, satellite image analysis, and GIS modeling, we developed a high-resolution map of malaria entomological inoculation rates (EIR) in The Gambia, West Africa. The analyses are based on the variation in exposure to malaria parasites experienced in 48 villages in 1996 and 21 villages in 1997. The entomological inoculation rate (EIR) varied from 0 to 166 infective bites per person per rainy season. Detailed field surveys identified the major Anopheles gambiae s.l. breeding habitats. These habitats were mapped by classification of a LANDSAT TM satellite image with an overall accuracy of 85%. Village EIRs decreased as a power function based on the breeding areas size and proximity. We use this relationship and the breeding habitats to map the variation in EIR over the entire 2500-km(2) study area.

  4. Habitat characterization and spatial distribution of Anopheles sp. mosquito larvae in Dar es Salaam (Tanzania) during an extended dry period

    PubMed Central

    Sattler, Michael A; Mtasiwa, Deo; Kiama, Michael; Premji, Zul; Tanner, Marcel; Killeen, Gerry F; Lengeler, Christian

    2005-01-01

    Introduction By 2030, more than 50% of the African population will live in urban areas. Controlling malaria reduces the disease burden and further improves economic development. As a complement to treated nets and prompt access to treatment, measures targeted against the larval stage of Anopheles sp. mosquitoes are a promising strategy for urban areas. However, a precise knowledge of the geographic location and potentially of ecological characteristics of breeding sites is of major importance for such interventions. Methods In total 151 km2 of central Dar es Salaam, the biggest city of Tanzania, were systematically searched for open mosquito breeding sites. Ecologic parameters, mosquito larvae density and geographic location were recorded for each site. Logistic regression analysis was used to determine the key ecological factors explaining the different densities of mosquito larvae. Results A total of 405 potential open breeding sites were examined. Large drains, swamps and puddles were associated with no or low Anopheles sp. larvae density. The probability of Anopheles sp. larvae to be present was reduced when water was identified as "turbid". Small breeding sites were more commonly colonized by Anopheles sp. larvae. Further, Anopheles gambiae s.l. larvae were found in highly organically polluted habitats. Conclusions Clear ecological characteristics of the breeding requirements of Anopheles sp. larvae could not be identified in this setting. Hence, every stagnant open water body, including very polluted ones, have to be considered as potential malaria vector breeding sites. PMID:15649333

  5. Cyt1A from Bacillus thuringiensis Synergizes Activity of Bacillus sphaericus against Aedes aegypti (Diptera: Culicidae)

    PubMed Central

    Wirth, Margaret C.; Federici, Brian A.; Walton, William E.

    2000-01-01

    Bacillus sphaericus is a mosquitocidal bacterium recently developed as a commercial larvicide that is used worldwide to control pestiferous and vector mosquitoes. Whereas B. sphaericus is highly active against larvae of Culex and Anopheles mosquitoes, it is virtually nontoxic to Aedes aegypti, an important vector species. In the present study, we evaluated the capacity of the cytolytic protein Cyt1A from Bacillus thuringiensis subsp. israelensis to enhance the toxicity of B. sphaericus toward A. aegypti. Various combinations of these two materials were evaluated, and all were highly toxic. A ratio of 10:1 of B. sphaericus to Cyt1A was 3,600-fold more toxic to A. aegypti than B. sphaericus alone. Statistical analysis showed this high activity was due to synergism between the Cyt1A toxin and B. sphaericus. These results suggest that Cyt1A could be useful in expanding the host range of B. sphaericus. PMID:10698776

  6. Evaluation of a eucalyptus-based repellent against Anopheles spp. in Tanzania.

    PubMed

    Trigg, J K

    1996-06-01

    A eucalyptus-based insect repellent (PMD) with the principal active ingredient p-menthane-3,8-diol was evaluated in the field in comparison with deet. In human landing catches in Tanzania, 3 formulations of PMD were tested against Anopheles gambiae and An. funestus. Repellents, applied to the legs and feet at doses chosen as used in practice, gave complete protection from biting for between 6 and 7.75 h, depending upon the formulation type, with no significant difference between PMD and deet in terms of efficacy and duration of protection. PMID:8827599

  7. Sampling of An.gambiae s.s mosquitoes using Limburger cheese, heat and moisture as baits in a homemade trap

    PubMed Central

    2011-01-01

    Background Ample evidence has shown that odour baited traps are likely to provide an objective monitoring tool for the host-seeking fraction of mosquito vectors of diseases like malaria and bancroftian filariasis. Such traps could eventually become part of primary healthcare systems used to study the vector biology and epidemiology of mosquito-borne diseases. I hereby, report a study that sampled Anopheles gambiae sensu stricto mosquitoes in a screen house using a homemade trap baited with a combination of Limburger cheese and moisture, Limburger cheese and heat, or Limburger cheese, moisture and heat. Findings Tests on the efficacy of the developed trap to sample An. gambiae s.s, mosquitoes using Limburger cheese, moisture and heat as baits were carried out in a screen house measuring 11.4 × 7.1 × 2.8 m. The studies were done in three phases. In the first phase the efficacy of the trap to sample An. gambiae s.s. using odour and moisture was tested. The second phase was to test the efficacy of the trap to sample An. gambiae s.s. using Limburger cheese and heat. In the third phase a combination of Limburger cheese, moisture and heat was tested. Tests were carried out for 27 consecutive nights. The designed trap collected a total of 59 An. gambiae s.s. in three trials. The trap baited with Limburger cheese and moisture collected 7 An. gambiae s.s in 7 days. The mean catch per day was 1. The trap baited with Limburger cheese and heat collected zero An. gambiae s.s in 11 days. The mean catch per day was therefore 0. The trap baited with Limburger cheese, moisture and heat collected 52 mosquitoes in 27 days and the mean catch was 1.93. Conclusions This study indicates that a non-electric fan driven trap baited with a combination of Limburger cheese, heat and moisture has a potential as an effective sampling tool for the malaria vector, Anopheles gambiae s.s. However, further optimization studies would be necessary. PMID:21835032

  8. GENETIC ISOLATION WITHIN THE MALARIA MOSQUITO ANOPHELES MELAS

    PubMed Central

    Deitz, Kevin C; Athrey, Giri; Reddy, Michael R; Overgaard, Hans J; Matias, Abrahan; Jawara, Musa; della Torre, Alessandra; Petrarca, Vincenzo; Pinto, Joao; Kiszewski, Anthony; Kengne, Pierre; Costantini, Carlo; Caccone, Adalgisa; Slotman, Michel A

    2014-01-01

    Anopheles melas is a brackish water-breeding member of the An. gambiae complex that is distributed along the coast of West Africa and is a major malaria vector within its range. Because little is known about the population structure of this species, we analyzed 15 microsatellite markers and 1,161 bp of mtDNA in 11 An. melas populations collected throughout its range. Compared to its sibling species An. gambiae, An. melas populations have a high level of genetic differentiation between them, representing its patchy distribution due to its fragmented larval habitat which is associated with mangroves and salt marsh grass. Populations clustered into three distinct groups representing Western Africa, Southern Africa, and Bioko Island populations that appear to be mostly isolated. Fixed differences in the mtDNA are present between all three clusters, and a Bayesian clustering analysis of the microsatellite data found no evidence for migration from mainland to Bioko Island populations, and little migration was evident between the Southern to the Western cluster. Surprisingly, mtDNA divergence between the three An. melas clusters is on par with levels of divergence between other species of the An. gambiae complex, and no support for monophyly was observed in a maximum likelihood phylogenetic analysis. Finally, an Approximate Bayesian Analysis of microsatellite data indicates that Bioko Island An. melas populations were connected to the mainland populations in the past, but became isolated, presumably when sea levels rose after the last glaciation period (≥10,000-11,000 years ago). This study has exposed species level genetic divergence within An. melas, and also has implications for control of this malaria vector. PMID:22882458

  9. Microevolution of Aedes aegypti

    PubMed Central

    Louise, Caroline; Vidal, Paloma Oliveira; Suesdek, Lincoln

    2015-01-01

    Scientific research into the epidemiology of dengue frequently focuses on the microevolution and dispersion of the mosquito Aedes aegypti. One of the world’s largest urban agglomerations infested by Ae. aegypti is the Brazilian megalopolis of Sao Paulo, where >26,900 cases of dengue were reported until June 2015. Unfortunately, the dynamics of the genetic variability of Ae. aegypti in the Sao Paulo area have not been well studied. To reduce this knowledge gap, we assessed the morphogenetic variability of a population of Ae. aegypti from a densely urbanised neighbourhood of Sao Paulo. We tested if allelic patterns could vary over a short term and if wing shape could be a predictor of the genetic variation. Over a period of 14 months, we examined the variation of genetic (microsatellites loci) and morphological (wing geometry) markers in Ae. aegypti. Polymorphisms were detected, as revealed by the variability of 20 microsatellite loci (115 alleles combined; overall Fst = 0.0358) and 18 wing landmarks (quantitative estimator Qst = 0.4732). These levels of polymorphism are higher than typically expected to an exotic species. Allelic frequencies of the loci changed over time and temporal variation in the wing shape was even more pronounced, permitting high reclassification levels of chronological samples. In spite of the fact that both markers underwent temporal variation, no correlation was detected between their dynamics. We concluded that microevolution was detected despite the short observational period, but the intensities of change of the markers were discrepant. Wing shape failed from predicting allelic temporal variation. Possibly, natural selection (Qst>Fst) or variance of expressivity of wing phenotype are involved in this discrepancy. Other possibly influential factors on microevolution of Ae. aegypti are worth searching. Additionally, the implications of the rapid evolution and high polymorphism of this mosquito vector on the efficacy of control methods

  10. Microevolution of Aedes aegypti.

    PubMed

    Louise, Caroline; Vidal, Paloma Oliveira; Suesdek, Lincoln

    2015-01-01

    Scientific research into the epidemiology of dengue frequently focuses on the microevolution and dispersion of the mosquito Aedes aegypti. One of the world's largest urban agglomerations infested by Ae. aegypti is the Brazilian megalopolis of Sao Paulo, where >26,900 cases of dengue were reported until June 2015. Unfortunately, the dynamics of the genetic variability of Ae. aegypti in the Sao Paulo area have not been well studied. To reduce this knowledge gap, we assessed the morphogenetic variability of a population of Ae. aegypti from a densely urbanised neighbourhood of Sao Paulo. We tested if allelic patterns could vary over a short term and if wing shape could be a predictor of the genetic variation. Over a period of 14 months, we examined the variation of genetic (microsatellites loci) and morphological (wing geometry) markers in Ae. aegypti. Polymorphisms were detected, as revealed by the variability of 20 microsatellite loci (115 alleles combined; overall Fst = 0.0358) and 18 wing landmarks (quantitative estimator Qst = 0.4732). These levels of polymorphism are higher than typically expected to an exotic species. Allelic frequencies of the loci changed over time and temporal variation in the wing shape was even more pronounced, permitting high reclassification levels of chronological samples. In spite of the fact that both markers underwent temporal variation, no correlation was detected between their dynamics. We concluded that microevolution was detected despite the short observational period, but the intensities of change of the markers were discrepant. Wing shape failed from predicting allelic temporal variation. Possibly, natural selection (Qst>Fst) or variance of expressivity of wing phenotype are involved in this discrepancy. Other possibly influential factors on microevolution of Ae. aegypti are worth searching. Additionally, the implications of the rapid evolution and high polymorphism of this mosquito vector on the efficacy of control methods have

  11. Complete Dosage Compensation in Anopheles stephensi and the Evolution of Sex-Biased Genes in Mosquitoes

    PubMed Central

    Jiang, Xiaofang; Biedler, James K.; Qi, Yumin; Hall, Andrew Brantley; Tu, Zhijian

    2015-01-01

    Complete dosage compensation refers to hyperexpression of the entire X or Z chromosome in organisms with heterogametic sex chromosomes (XY male or ZW female) in order to compensate for having only one copy of the X or Z chromosome. Recent analyses suggest that complete dosage compensation, as in Drosophila melanogaster, may not be the norm. There has been no systematic study focusing on dosage compensation in mosquitoes. However, analysis of dosage compensation in Anopheles mosquitoes provides opportunities for evolutionary insights, as the X chromosome of Anopheles and that of its Dipteran relative, D. melanogaster formed independently from the same ancestral chromosome. Furthermore, Culicinae mosquitoes, including the Aedes genus, have homomorphic sex-determining chromosomes, negating the need for dosage compensation. Thus, Culicinae genes provide a rare phylogenetic context to investigate dosage compensation in Anopheles mosquitoes. Here, we performed RNA-seq analysis of male and female samples of the Asian malaria mosquito Anopheles stephensi and the yellow fever mosquito Aedes aegypti. Autosomal and X-linked genes in An. stephensi showed very similar levels of expression in both males and females, indicating complete dosage compensation. The uniformity of average expression levels of autosomal and X-linked genes remained when An. stephensi gene expression was normalized by that of their Ae. aegypti orthologs, strengthening the finding of complete dosage compensation in Anopheles. In addition, we comparatively analyzed the differentially expressed genes between adult males and adult females in both species, investigated sex-biased gene chromosomal distribution patterns in An. stephensi and provided three examples where gene duplications may have enabled the acquisition of sex-specific expression during mosquito evolution. PMID:26078263

  12. Evaluation of environmental data for identification of Anopheles (Diptera: Culicidae) aquatic larval habitats in Kisumu and Malindi, Kenya.

    PubMed

    Jacob, Benjamin G; Arheart, Kristopher L; Griffith, Daniel A; Mbogo, Charles M; Githeko, Andrew K; Regens, James L; Githure, John I; Novak, Robert; Beier, John C

    2005-09-01

    This research evaluates the extent to which use of environmental data acquired from field and satellite surveys enhances predictions of urban mosquito counts. Mosquito larval habitats were sampled, and multispectral thermal imager (MTI) satellite data in the visible spectrum at 5-m resolution were acquired for Kisumu and Malindi, Kenya, during February and March 2001. All entomological parameters were collected from January to May 2001, June to August 2002, and June to August 2003. In a Poisson model specification, for Anopheles funestus Giles, shade was the best predictor, whereas substrate was the best predictor for Anopheles gambiae, and vegetation for Anopheles arabensis Patton. The top predictors found with a logistic regression model specification were habitat size for An. gambiae Giles, pollution for An. arabensis, and shade for An. funestus. All other coefficients for canopy, debris, habitat nature, permanency, emergent plants, algae, pollution, turbidity, organic materials, all MTI waveband frequencies, distance to the nearest house, distance to the nearest domestic animal, and all land use land cover changes were nonsignificant. MTI data at 5-m spatial resolution do not have an additional predictive value for mosquito counts when adjusted for field-based ecological data. PMID:16365996

  13. A New Role for an Old Antimicrobial: Lysozyme c-1 Can Function to Protect Malaria Parasites in Anopheles Mosquitoes

    PubMed Central

    Li, Bin; Luckhart, Shirley; Li, Jianyong; Paskewitz, Susan M.

    2011-01-01

    Background Plasmodium requires an obligatory life stage in its mosquito host. The parasites encounter a number of insults while journeying through this host and have developed mechanisms to avoid host defenses. Lysozymes are a family of important antimicrobial immune effectors produced by mosquitoes in response to microbial challenge. Methodology/Principal Findings A mosquito lysozyme was identified as a protective agonist for Plasmodium. Immunohistochemical analyses demonstrated that Anopheles gambiae lysozyme c-1 binds to oocysts of Plasmodium berghei and Plasmodium falciparum at 2 and 5 days after infection. Similar results were observed with Anopheles stephensi and P. falciparum, suggesting wide occurrence of this phenomenon across parasite and vector species. Lysozyme c-1 did not bind to cultured ookinetes nor did recombinant lysozyme c-1 affect ookinete viability. dsRNA-mediated silencing of LYSC-1 in Anopheles gambiae significantly reduced the intensity and the prevalence of Plasmodium berghei infection. We conclude that this host antibacterial protein directly interacts with and facilitates development of Plasmodium oocysts within the mosquito. Conclusions/Significance This work identifies mosquito lysozyme c-1 as a positive mediator of Plasmodium development as its reduction reduces parasite load in the mosquito host. These findings improve our understanding of parasite development and provide a novel target to interrupt parasite transmission to human hosts. PMID:21573077

  14. Evaluation of environmental data for identification of Anopheles (Diptera: Culicidae) aquatic larval habitats in Kisumu and Malindi, Kenya.

    PubMed

    Jacob, Benjamin G; Arheart, Kristopher L; Griffith, Daniel A; Mbogo, Charles M; Githeko, Andrew K; Regens, James L; Githure, John I; Novak, Robert; Beier, John C

    2005-09-01

    This research evaluates the extent to which use of environmental data acquired from field and satellite surveys enhances predictions of urban mosquito counts. Mosquito larval habitats were sampled, and multispectral thermal imager (MTI) satellite data in the visible spectrum at 5-m resolution were acquired for Kisumu and Malindi, Kenya, during February and March 2001. All entomological parameters were collected from January to May 2001, June to August 2002, and June to August 2003. In a Poisson model specification, for Anopheles funestus Giles, shade was the best predictor, whereas substrate was the best predictor for Anopheles gambiae, and vegetation for Anopheles arabensis Patton. The top predictors found with a logistic regression model specification were habitat size for An. gambiae Giles, pollution for An. arabensis, and shade for An. funestus. All other coefficients for canopy, debris, habitat nature, permanency, emergent plants, algae, pollution, turbidity, organic materials, all MTI waveband frequencies, distance to the nearest house, distance to the nearest domestic animal, and all land use land cover changes were nonsignificant. MTI data at 5-m spatial resolution do not have an additional predictive value for mosquito counts when adjusted for field-based ecological data.

  15. Larval competition between An. coluzzii and An. gambiae in insectary and semi-field conditions in Burkina Faso

    PubMed Central

    Brossette, Lou; Mamaï, Wadaka; Dabiré, Roch; Simard, Frédéric

    2014-01-01

    Competition in mosquito larvae is common and different ecological context could change competitive advantage between species. Here, larval competition between the widely sympatric African malaria mosquitoes, Anopheles coluzzii and Anopheles gambiae were investigated in controlled insectary conditions using individuals from laboratory colonies and under ambient conditions using wild mosquitoes in a semi-field enclosure in western Burkina Faso. Larvae of both species were reared in trays at the same larval density and under the same feeding regimen in either single-species or mixed-species populations at varying species ratios reflecting 0, 25, 50 and 75% of competitor species. In the insectaries, where environmental variations are controlled, larvae of the An. coluzzii colony developed faster and with lower mortality than larvae of the An. gambiae colony (8.8±0.1 days and 21±3% mortality vs. 9.5±0.1 days and 32±3% mortality, respectively). Although there was no significant effect of competition on these phenotypic traits in any species, there was a significant trend for higher fitness of the An. coluzzii colony when competing with An. gambiae under laboratory conditions (i.e., lower development time and increased wing length at emergence, Cuzik’s tests, P<0.05). In semi-field experiments, competition affected the life history traits of both species in a different way. Larvae of An. gambiae tended to reduce development time when in competition with An. coluzzii (Cuzick’s test, P=0.002) with no impact either on mortality or size at emergence. On the other hand, An. coluzzii showed a significant trend for reduced larval mortality with increasing competition pressure (Cuzick’s test, P=0.037) and production of smaller females when grown together with An. gambiae (Cuzick’s test, P=0.002). Our results hence revealed that competitive interactions between larvae of the two species are context dependent. They further call for caution when exploring ecological

  16. Genome-Wide Divergence in the West-African Malaria Vector Anopheles melas.

    PubMed

    Deitz, Kevin C; Athrey, Giridhar A; Jawara, Musa; Overgaard, Hans J; Matias, Abrahan; Slotman, Michel A

    2016-01-01

    Anopheles melas is a member of the recently diverged An. gambiae species complex, a model for speciation studies, and is a locally important malaria vector along the West-African coast where it breeds in brackish water. A recent population genetic study of An. melas revealed species-level genetic differentiation between three population clusters. An. melas West extends from The Gambia to the village of Tiko, Cameroon. The other mainland cluster, An. melas South, extends from the southern Cameroonian village of Ipono to Angola. Bioko Island, Equatorial Guinea An. melas populations are genetically isolated from mainland populations. To examine how genetic differentiation between these An. melas forms is distributed across their genomes, we conducted a genome-wide analysis of genetic differentiation and selection using whole genome sequencing data of pooled individuals (Pool-seq) from a representative population of each cluster. The An. melas forms exhibit high levels of genetic differentiation throughout their genomes, including the presence of numerous fixed differences between clusters. Although the level of divergence between the clusters is on a par with that of other species within the An. gambiae complex, patterns of genome-wide divergence and diversity do not provide evidence for the presence of pre- and/or postmating isolating mechanisms in the form of speciation islands. These results are consistent with an allopatric divergence process with little or no introgression. PMID:27466271

  17. Linear and spatial organization of polytene chromosomes of the African malaria mosquito Anopheles funestus.

    PubMed Central

    Sharakhov, I V; Sharakhova, M V; Mbogo, C M; Koekemoer, L L; Yan, G

    2001-01-01

    Anopheles funestus Giles is one of the major malaria vectors in Africa, but little is known about its genetics. Lack of a cytogenetic map characterized by regions has hindered the progress of genetic research with this important species. This study developed a cytogenetic map of An. funestus using ovarian nurse cell polytene chromosomes. We demonstrate an important application with the cytogenetic map for characterizing various chromosomal inversions for specimens collected from coastal Kenya. The linear and spatial organization of An. funestus polytene chromosomes was compared with the best-studied malaria mosquito, An. gambiae Giles. Comparisons of chromosome morphology between the two species have revealed that the most extensive chromosomal rearrangement occurs in pericentromeric heterochromatin of autosomes. Differences in pericentromeric heterochromatin types correlate with nuclear organization differences between An. funestus and An. gambiae. Attachments of chromosomes to the nuclear envelope strongly depend on the presence of diffusive beta-heterochromatin. Thus, An. funestus and An. gambiae exhibit species-specific characteristics in chromosome-linear and -spatial organizations. PMID:11560898

  18. Genome-Wide Divergence in the West-African Malaria Vector Anopheles melas

    PubMed Central

    Deitz, Kevin C.; Athrey, Giridhar A.; Jawara, Musa; Overgaard, Hans J.; Matias, Abrahan; Slotman, Michel A.

    2016-01-01

    Anopheles melas is a member of the recently diverged An. gambiae species complex, a model for speciation studies, and is a locally important malaria vector along the West-African coast where it breeds in brackish water. A recent population genetic study of An. melas revealed species-level genetic differentiation between three population clusters. An. melas West extends from The Gambia to the village of Tiko, Cameroon. The other mainland cluster, An. melas South, extends from the southern Cameroonian village of Ipono to Angola. Bioko Island, Equatorial Guinea An. melas populations are genetically isolated from mainland populations. To examine how genetic differentiation between these An. melas forms is distributed across their genomes, we conducted a genome-wide analysis of genetic differentiation and selection using whole genome sequencing data of pooled individuals (Pool-seq) from a representative population of each cluster. The An. melas forms exhibit high levels of genetic differentiation throughout their genomes, including the presence of numerous fixed differences between clusters. Although the level of divergence between the clusters is on a par with that of other species within the An. gambiae complex, patterns of genome-wide divergence and diversity do not provide evidence for the presence of pre- and/or postmating isolating mechanisms in the form of speciation islands. These results are consistent with an allopatric divergence process with little or no introgression. PMID:27466271

  19. Plasmodium falciparum infection rates for some Anopheles spp. from Guinea-Bissau, West Africa

    PubMed Central

    Sanford, Michelle R.; Cornel, Anthony J.; Nieman, Catelyn C.; Dinis, Joao; Marsden, Clare D.; Weakley, Allison M.; Han, Sarah; Rodrigues, Amabelia; Lanzaro, Gregory C.; Lee, Yoosook

    2014-01-01

    Presence of Plasmodium falciparum circumsporozoite protein (CSP) was detected by enzyme linked immunosorbent assay (ELISA) in a sample of Anopheles gambiae s.s., A. melas and A. pharoensis collected in Guinea-Bissau during October and November 2009. The percentage of P. falciparum infected samples (10.2% overall; confidence interval (CI): 7.45-13.6%) was comparable to earlier studies from other sites in Guinea-Bissau (9.6-12.4%). The majority of the specimens collected were identified as A. gambiae which had an individual infection rate of 12.6 % (CI: 8.88-17.6) across collection sites. A small number of specimens of A. coluzzii, A. coluzzii x A. gambiae hybrids, A. melas and A. pharoensis were collected and had infection rates of 4.3% (CI:0.98-12.4), 4.1% (CI:0.35-14.5), 11.1% (CI:1.86-34.1) and 33.3% (CI:9.25-70.4) respectively. Despite being present in low numbers in indoor collections, the exophilic feeding behaviors of A. melas (N=18) and A. pharoensis (N=6) and high infection rates observed in this survey suggest falciparum-malaria transmission potential outside of the protection of bed nets. PMID:25383188

  20. Aedes aegypti genomics.

    PubMed

    Severson, David W; Knudson, Dennis L; Soares, Marcelo B; Loftus, Brendan J

    2004-07-01

    The mosquito, Aedes aegypti, is the primary, worldwide arthropod vector for the yellow fever and dengue viruses. As it is also one of the most tractable mosquito species for laboratory studies, it has been and remains one of the most intensively studied arthropod species. This has resulted in the development of detailed genetic and physical maps for Ae. aegypti and considerable insight into its genome organization. The research community is well-advanced in developing important molecular tools that will facilitate a whole genome sequencing effort. This includes generation of BAC clone end sequences, physical mapping of selected BAC clones and generation of EST sequences. Whole genome sequence information for Ae. aegypti will provide important insight into mosquito chromosome evolution and allow for the identification of genes and gene function. These functions may be common to all mosquitoes or perhaps unique to individual species, possibly specific to host-seeking and blood-feeding behaviors, as well as the innate immune response to pathogens encountered during blood-feeding. This information will be invaluable to the global effort to develop novel strategies for preventing arthropod-borne disease transmission.

  1. Anopheles ziemanni a locally important malaria vector in Ndop health district, north west region of Cameroon

    PubMed Central

    2014-01-01

    Background Malaria transmission in Cameroon is mediated by a plethora of vectors that are heterogeneously distributed across the country depending on the biotope. To effectively guide malaria control operations, regular update on the role of local Anopheles species is essential. Therefore, an entomological survey was conducted between August 2010 and May 2011 to evaluate the role of the local anopheline population in malaria transmission in three villages of the Ndop health district in the northwest region of Cameroon where malaria is holoendemic, as a means to acquiring evidence based data for improved vector intervention. Methods Mosquitoes were sampled both indoor and outdoor for four consecutive nights in each locality during each month of survey. Sampling was done by the human landing catch method on volunteers. Anopheles species were identified morphologically and their ovaries randomly dissected for parity determination. Infection with Plasmodium falciparum was detected by Circumsporozoite protein ELISA. Members of An. gambiae complex were further identified to molecular level by PCR and RFLP PCR. Results An. ziemanni was the main malaria vector and whether outdoor or indoor. The man biting rate for the vectors ranged from 6.75 to 8.29 bites per person per night (b/p/n). The entomological inoculation rate for this vector species was 0.0278 infectious bites per person per night (ib/p/n) in Mbapishi, 0.034 ib/p/n in Mbafuh, and 0.063 ib/p/n in Backyit. These were by far greater than that for An. gambiae. No difference was observed in the parity rate of these two vectors. PCR analysis revealed the presence of only An. colluzzi (M- form). Conclusions An. ziemanni is an important local malaria vector in Ndop health district. The findings provide useful baseline information on the anopheles species composition, their distribution and role in malaria transmission that would guide the implementation of integrated vector management strategies in the locality. PMID

  2. Characterization and expression analysis of gene encoding heme peroxidase HPX15 in major Indian malaria vector Anopheles stephensi (Diptera: Culicidae).

    PubMed

    Kajla, Mithilesh; Kakani, Parik; Choudhury, Tania Pal; Gupta, Kuldeep; Gupta, Lalita; Kumar, Sanjeev

    2016-06-01

    The interaction of mosquito immune system with Plasmodium is critical in determining the vector competence. Thus, blocking the crucial mosquito molecules that regulate parasite development might be effective in controlling the disease transmission. In this study, we characterized a full-length AsHPX15 gene from the major Indian malaria vector Anopheles stephensi. This gene is true ortholog of Anopheles gambiae heme peroxidase AgHPX15 (AGAP013327), which modulates midgut immunity and regulates Plasmodium falciparum development. We found that AsHPX15 is highly induced in mosquito developmental stages and blood fed midguts. In addition, this is a lineage-specific gene that has identical features and 65-99% amino acids identity with other HPX15 genes present in eighteen worldwide-distributed anophelines. We discuss that the conserved HPX15 gene might serve as a common target to manipulate mosquito immunity and arresting Plasmodium development inside the vector host.

  3. A simple and affordable membrane-feeding method for Aedes aegpyti and Anopheles minimus (Diptera: Culicidae).

    PubMed

    Finlayson, Catherine; Saingamsook, Jassada; Somboon, Pradya

    2015-12-01

    This study developed an artificial feeding (AF) method to replace direct host feeding (DHF) for the maintenance of Aedes aegypti and Anopheles minimus mosquito colonies. The procedure can be adopted by all laboratories due to its simple and affordable materials and design. The apparatus consists of heparinized cow blood contained in a 5cm diameter glass petri dish with 5cm(2) Parafilm M (Bemis(®)) stretched thinly over the top, with a pre-heated bag of vegetable oil placed underneath to keep the blood warm. Both parts are contained within an insulated Styrofoam™ box with a hole in the lid for mosquitoes to access the membrane. Mosquitoes are fed by AF for 15min at a time. Feeding rate and fecundity of Ae. aegypti mosquitoes feeding on the AF device were compared to those feeding on a live rat (DHF(r)), and of Anopheles minimus mosquitoes feeding on the AF device compared to those feeding on a human arm (DHF(h)). Aedes aegypti mosquitoes fed by AF or DHF(r) had similar feeding rates (38.2±21.5% and 35.7±18.2%, respectively) and overall egg production (1.5% difference). Anopheles minimus mosquitoes fed by the AF method had a lower feeding rate (52.0±1.0% for AF compared to 70.7±20.2% for DHF(h)) and overall egg production (40% reduction compared to DHF(h)). However, the number of eggs produced by AF-fed mosquitoes (1808 eggs per 100 mosquitoes) was still sufficient for colony maintenance, and with increased feeding time both parameters are expected to increase. Reduced feeding rate and overall egg production was observed when Ae. aegypti mosquitoes were fed on blood refrigerated for over two weeks. In conclusion, an AF device has been developed which can replace DHF for Ae. aegypti and An. minimus colony maintenance when using blood refrigerated for a maximum of two weeks. PMID:26440474

  4. A simple and affordable membrane-feeding method for Aedes aegpyti and Anopheles minimus (Diptera: Culicidae).

    PubMed

    Finlayson, Catherine; Saingamsook, Jassada; Somboon, Pradya

    2015-12-01

    This study developed an artificial feeding (AF) method to replace direct host feeding (DHF) for the maintenance of Aedes aegypti and Anopheles minimus mosquito colonies. The procedure can be adopted by all laboratories due to its simple and affordable materials and design. The apparatus consists of heparinized cow blood contained in a 5cm diameter glass petri dish with 5cm(2) Parafilm M (Bemis(®)) stretched thinly over the top, with a pre-heated bag of vegetable oil placed underneath to keep the blood warm. Both parts are contained within an insulated Styrofoam™ box with a hole in the lid for mosquitoes to access the membrane. Mosquitoes are fed by AF for 15min at a time. Feeding rate and fecundity of Ae. aegypti mosquitoes feeding on the AF device were compared to those feeding on a live rat (DHF(r)), and of Anopheles minimus mosquitoes feeding on the AF device compared to those feeding on a human arm (DHF(h)). Aedes aegypti mosquitoes fed by AF or DHF(r) had similar feeding rates (38.2±21.5% and 35.7±18.2%, respectively) and overall egg production (1.5% difference). Anopheles minimus mosquitoes fed by the AF method had a lower feeding rate (52.0±1.0% for AF compared to 70.7±20.2% for DHF(h)) and overall egg production (40% reduction compared to DHF(h)). However, the number of eggs produced by AF-fed mosquitoes (1808 eggs per 100 mosquitoes) was still sufficient for colony maintenance, and with increased feeding time both parameters are expected to increase. Reduced feeding rate and overall egg production was observed when Ae. aegypti mosquitoes were fed on blood refrigerated for over two weeks. In conclusion, an AF device has been developed which can replace DHF for Ae. aegypti and An. minimus colony maintenance when using blood refrigerated for a maximum of two weeks.

  5. Gene Flow, Subspecies Composition, and Dengue Virus-2 Susceptibility among Aedes aegypti Collections in Senegal

    PubMed Central

    Sylla, Massamba; Bosio, Christopher; Urdaneta-Marquez, Ludmel; Ndiaye, Mady; Black, William C.

    2009-01-01

    Background Aedes aegypti, the “yellow fever mosquito”, is the primary vector to humans of the four serotypes of dengue viruses (DENV1-4) and yellow fever virus (YFV) and is a known vector of Chikungunya virus. There are two recognized subspecies of Ae. aegypti sensu latu (s.l.): the presumed ancestral form, Ae. aegypti formosus (Aaf), a primarily sylvan mosquito in sub-Saharan Africa, and Ae. aegypti aegypti (Aaa), found globally in tropical and subtropical regions typically in association with humans. The designation of Ae. aegypti s.l. subspecies arose from observations made in East Africa in the late 1950s that the frequency of pale “forms” of Ae. aegypti was higher in populations in and around human dwellings than in those of the nearby bush. But few studies have been made of Ae. aegypti s.l. in West Africa. To address this deficiency we have been studying the population genetics, subspecies composition and vector competence for DENV-2 of Ae. aegypti s.l. in Senegal. Methods and Findings A population genetic analysis of gene flow was conducted among 1,040 Aedes aegypti s.l. from 19 collections distributed across the five phytogeographic regions of Senegal. Adults lacking pale scales on their first abdominal tergite were classified as Aedes aegypti formosus (Aaf) following the original description of the subspecies and the remainder were classified as Aedes aegypti aegypti (Aaa). There was a clear northwest–southeast cline in the abundance of Aaa and Aaf. Collections from the northern Sahelian region contained only Aaa while southern Forest gallery collections contained only Aaf. The two subspecies occurred in sympatry in four collections north of the Gambia in the central Savannah region and Aaa was a minor component of two collections from the Forest gallery area. Mosquitoes from 11 collections were orally challenged with DENV-2 virus. In agreement with the early literature, Aaf had significantly lower vector competence than Aaa. Among pure Aaa

  6. Flavivirus susceptibility in Aedes aegypti.

    PubMed

    Black, William C; Bennett, Kristine E; Gorrochótegui-Escalante, Norma; Barillas-Mury, Carolina V; Fernández-Salas, Ildefonso; de Lourdes Muñoz, María; Farfán-Alé, José A; Olson, Ken E; Beaty, Barry J

    2002-01-01

    Aedes aegypti is the primary vector of yellow fever (YF) and dengue fever (DF) flaviviruses worldwide. In this review we focus on past and present research on genetic components and environmental factors in Aedes aegypti that appear to control flavivirus transmission. We review genetic relationships among Ae. aegypti populations throughout the world and discuss how variation in vector competence is correlated with overall genetic differences among populations. We describe current research into how genetic and environmental factors jointly affect distribution of vector competence in natural populations. Based on this information, we propose a population genetic model for vector competence and discuss our recent progress in testing this model. We end with a discussion of approaches being taken to identify the genes that may control flavivirus susceptibility in Ae. aegypti.

  7. Elimination of Lymphatic Filariasis in The Gambia

    PubMed Central

    Rebollo, Maria P.; Sambou, Sana Malang; Thomas, Brent; Biritwum, Nana-Kwadwo; Jaye, Momodou C.; Kelly-Hope, Louise; Escalada, Alba Gonzalez; Molyneux, David H.; Bockarie, Moses J.

    2015-01-01

    Background The prevalence of Wuchereria bancrofti, which causes lymphatic filariasis (LF) in The Gambia was among the highest in Africa in the 1950s. However, surveys conducted in 1975 and 1976 revealed a dramatic decline in LF endemicity in the absence of mass drug administration (MDA). The decline in prevalence was partly attributed to a significant reduction in mosquito density through the widespread use of insecticidal nets. Based on findings elsewhere that vector control alone can interrupt LF, we asked the question in 2013 whether the rapid scale up in the use of insecticidal nets in The Gambia had interrupted LF transmission. Methodology/Principal Finding We present here the results of three independently designed filariasis surveys conducted over a period of 17 years (1997–2013), and involving over 6000 subjects in 21 districts across all administrative divisions in The Gambia. An immunochromatographic (ICT) test was used to detect W. bancrofti antigen during all three surveys. In 2001, tests performed on stored samples collected between 1997 and 2000, in three divisions, failed to show positive individuals from two divisions that were previously highly endemic for LF, suggesting a decline towards extinction in some areas. Results of the second survey conducted in 2003 showed that LF was no longer endemic in 16 of 21 districts surveyed. The 2013 survey used a WHO recommended LF transmission verification tool involving 3180 6–7 year-olds attending 60 schools across the country. We demonstrated that transmission of W. bancrofti has been interrupted in all 21 districts. Conclusions We conclude that LF transmission may have been interrupted in The Gambia through the extensive use of insecticidal nets for malaria control for decades. The growing evidence for the impact of malaria vector control activities on parasite transmission has been endorsed by WHO through a position statement in 2011 on integrated vector management to control malaria and LF. PMID

  8. Attractiveness of MM-X Traps Baited with Human or Synthetic Odor to Mosquitoes (Diptera: Culicidae) in The Gambia

    PubMed Central

    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

    2013-01-01

    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 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 + CO2 + 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. PMID:18047195

  9. Lactate dehydrogenase as a marker of Plasmodium infection in malaria vector Anopheles.

    PubMed

    Riandey, M F; Sannier, C; Peltre, G; Monteny, N; Cavaleyra, M

    1996-06-01

    Lactate dehydrogenase (Ldh) electrophoresis showed the presence of Plasmodium yoelii yoelii in Anopheles stephensi and An. gambiae. The Ldh appeared as an additional band (pLdh) whose activity was more intense with 3-acetyl pyridine adenine dinucleotide as coenzyme than with beta nicotin-amide adenine dinucleotide. Several allelic forms occurred both in the vector and the host. The isoelectric point of Ldh, similar in the vector and host, differed from those of Ldh from mosquito and mouse. The presence of pLdh was detected from the 2nd to the 28th day of infection. The pLdh appeared to be proportional to the number of sporozoites present in infected salivary glands. However, pLdh was not found in salivary glands or midguts, but it was detected in the rest of the corresponding mosquito. The origin and use of pLdh as a marker of Plasmodium in its vector is discussed. PMID:8827592

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

    PubMed

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

    1993-01-01

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

  11. Plasmodium falciparum infection increases Anopheles gambiae attraction to nectar sources and sugar uptake

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Plasmodium parasites are known to manipulate the behaviour of their vectors so as to enhance their transmission. However, it is unknown if this vector manipulation also affects mosquito-plant interaction and sugar uptake. Dual-choice olfactometer and probing assays were used to study plant seeking b...

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

    Technology Transfer Automated Retrieval System (TEKTRAN)

    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 ...

  13. A New Role of the Mosquito Complement-like Cascade in Male Fertility in Anopheles gambiae