Identification of Spiroplasma insolitum symbionts in Anopheles gambiae [version 1; referees: 2 approved, 1 not approved

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Sharon T. Chepkemoi

2017-09-01

Full Text Available Background: Insect symbionts have the potential to block the transmission of vector-borne diseases by their hosts. The advancement of a symbiont-based transmission blocking strategy for malaria requires the identification and study of Anopheles symbionts. Methods: High throughput 16S amplicon sequencing was used to profile the bacteria associated with Anopheles gambiae sensu lato and identify potential symbionts. The polymerase chain reaction (PCR with specific primers were subsequently used to monitor symbiont prevalence in field populations, as well as symbiont transmission patterns. Results: We report the discovery of the bacterial symbiont, Spiroplasma, in Anopheles gambiae in Kenya. We determine that geographically dispersed Anopheles gambiae populations in Kenya are infected with Spiroplasma at low prevalence levels. Molecular phylogenetics indicates that this Anopheles gambiae associated Spiroplasma is a member of the insolitum clade. We demonstrate that this symbiont is stably maternally transmitted across at least two generations and does not significantly affect the fecundity or egg to adult survival of its host. Conclusions: In diverse insect species, Spiroplasma has been found to render their host resistant to infection by pathogens. The identification of a maternally transmitted strain of Spiroplasma in Anopheles gambiae may therefore open new lines of investigation for the development of symbiont-based strategies for blocking malaria transmission.

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

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Voordouw Maarten J

2007-07-01

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

Pyrethroids and DDT tolerance of Anopheles gambiae s.l. from Sengerema District, an area of intensive pesticide usage in north-western Tanzania.

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Philbert, Anitha; Lyantagaye, Sylvester Leonard; Pradel, Gabriele; Ngwa, Che Julius; Nkwengulila, Gamba

2017-04-01

To assess the susceptibility status of malaria vectors to pyrethroids and dichlorodiphenyltrichloroethane (DDT), characterise the mechanisms underlying resistance and evaluate the role of agro-chemical use in resistance selection among malaria vectors in Sengerema agro-ecosystem zone, Tanzania. Mosquito larvae were collected from farms and reared to obtain adults. The susceptibility status of An. gambiae s.l. was assessed using WHO bioassay tests to permethrin, deltamethrin, lambdacyhalothrin, etofenprox, cyfluthrin and DDT. Resistant specimens were screened for knock-down resistance gene (kdr), followed by sequencing both Western and Eastern African variants. A gas chromatography-mass spectrophotometer (GC-MS) was used to determine pesticide residues in soil and sediments from mosquitoes' breeding habitats. Anopheles gambiae s.l. was resistant to all the insecticides tested. The population of Anopheles gambiae s.l was composed of Anopheles arabiensis by 91%. The East African kdr (L1014S) allele was found in 13 of 305 specimens that survived insecticide exposure, with an allele frequency from 0.9% to 50%. DDTs residues were found in soils at a concentration up to 9.90 ng/g (dry weight). The observed high resistance levels of An. gambiae s.l., the detection of kdr mutations and pesticide residues in mosquito breeding habitats demonstrate vector resistance mediated by pesticide usage. An integrated intervention through collaboration of agricultural, livestock and vector control units is vital. © 2017 John Wiley & Sons Ltd.

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

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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.

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

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S. Kibret

2010-12-01

Full Text Available 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

A reliable morphological method to assess the age of male Anopheles gambiae

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

2006-01-01

Background - Release of genetically-modified (GM) or sterile male mosquitoes for malaria control is hampered by inability to assess the age and mating history of free-living male Anopheles. Methods - Age and mating-related changes in the reproductive system of male Anopheles gambiae were quantified

Melanotic pathology and vertical transmission of the gut commensal Elizabethkingia meningoseptica in the major malaria vector Anopheles gambiae.

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Idir G Akhouayri

Full Text Available 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.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.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.

Molecular characterization of larval peripheral thermosensory responses of the malaria vector mosquito Anopheles gambiae.

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Chao Liu

Full Text Available Thermosensation provides vital inputs for the malaria vector mosquito, Anopheles gambiae which utilizes heat-sensitivity within a broad spectrum of behaviors, most notably, the localization of human hosts for blood feeding. In this study, we examine thermosensory behaviors in larval-stage An. gambiae, which as a result of their obligate aquatic habitats and importance for vectorial capacity, represents an opportunistic target for vector control as part of the global campaign to eliminate malaria. As is the case for adults, immature mosquitoes respond differentially to a diverse array of external heat stimuli. In addition, larvae exhibit a striking phenotypic plasticity in thermal-driven behaviors that are established by temperature at which embryonic development occurs. Within this spectrum, RNAi-directed gene-silencing studies provide evidence for the essential role of the Transient Receptor Potential sub-family A1 (TRPA1 channel in mediating larval thermal-induced locomotion and thermal preference within a discrete upper range of ambient temperatures.

Landscape movements of Anopheles gambiae malaria vector mosquitoes in rural Gambia.

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

2013-01-01

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. 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. 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.

The physicochemical and environmental factors affecting the distribution of Anopheles merus along the Kenyan coast.

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Kipyab, Pamela C; Khaemba, Battan M; Mwangangi, Joseph M; Mbogo, Charles M

2015-04-11

Members of the Anopheles gambiae complex are the main transmitters of malaria. Anopheles merus is a member of the complex found along the Kenyan coast because it breeds in saline waters. An entomological study was conducted in Garithe Malindi District, to investigate the physicochemical and environmental factors affecting the distribution of An. merus. Field and laboratory studies were used to investigate the breeding habitats of the subspecies. Mosquito larvae were sampled using standard dipping technique from small pockets of pools, ponds, hoof prints, road drain, wells and mangrove swamps found in Garithe. All 3(rd) and 4(th) instars of Anopheles larvae sampled were identified microscopically into species. A representative of Anopheles gambiae complex was then identified to specific sibling species using r-DNA PCR technique. The habitats were characterized based on temperature, conductivity, salinity, dissolved oxygen, total dissolved solids, pH, size, distance to nearest house, canopy coverage, surface debris, presence of algae, emergent plants, turbidity and habitat types. A total of 159 morphologically identified late stage instar Anopheles gambiae s.l larvae were selected for r-DNA analysis by PCR. Out of these, 60.4% (n = 96) were Anopheles merus, 8.8% (n = 14) were Anopheles arabiensis, 18.2% (n = 29) were Anopheles gambiae s.s and 12.6% (n = 20) were unknown. Using paired t-test (t (121) = -3.331, P = 0.001) a significantly high proportion of An. merus was observed in all habitats compared to An. arabiensis, and An. gambiae s. s. In habitat characterization, Pearson's correlation analysis test showed different parameters being associated with the occurrence of An. merus larvae in the different habitats sampled. Six out of the 55 correlation coefficients (10.9%) were statistically significant, suggesting non-random association between some pairs of variables. Those that had a significantly high positive correlation with An. merus

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

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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

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

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Knols Bart GJ

2007-05-01

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

Efektivitas Bacillus thuringiensis dalam Pengendalian Larva Nyamuk Anopheles sp.

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Citra Inneke Wibowo

2017-08-01

Full Text Available Nyamuk Anopheles sp adalah vektor penyakit malaria. Pengendalian vektor penyakit malaria dapat dilakukan secara biologis yaitu dengan menggunakan Bacillus thuringiensis. Tujuan penelitian adalah untuk mengetahui efektivitas konsentrasi Bacillus thuringiensis dalam pengendalian larva nyamuk Anopheles sp.Penelitian ini dilakukan secara eksperimental menggunakan Rancangan Acak Lengkap Faktorial (RAL Faktorial yang terdiri atas dua faktor yaitu konsentrasi Bacillus thuringiensis dan stadia larva Anopheles dengan pengulangan tiga kali.Perlakuan yang dicobakan adalahkonsentrasi Bacillus thuringiensis (A yang terdiri atas 5 taraf:A0: konsentrasi B.thuringiensis 0 CFU.mL-1, A1: konsentrasi B.thuringiensis 102 CFU.mL-1, A2: konsentrasi B.thuringiensis 104 CFU.mL-1, A3: konsentrasi B.thuringiensis 106CFU.mL-1, A4: konsentrasi B.thuringiensis 108CFU.mL-1. Perlakuan tahapan instar larva Anopheles sp. (B adalah sebagai berikut:B1: stadia larva instar I, B2: stadia larva instar II, B3: stadia larva instar III, B4: stadia larva instar IVsehingga terdapat 60 satuan percobaan. Hasil penelitian  menunjukkan konsentrasi B. thuringiensis isolat CK dan IPB CC yang paling berpengaruh dalam pengendalian larva Anopheles sp adalah 108 CFU.mL-1 . Instar larva yang paling peka terhadap B. thuringiensis isolat IPB CC adalah instar I dan II sedangkan instar yang peka terhadap isolat CK adalah instar II, Perlakuan konsentrasi isolat B. thuringiensis dan tingkat instar larva yang paling baik dalam pengendalian larva Anopheles sp. adalah 108 CFU.mL-1, dan instar I dan II.

Larvicidal effect of disinfectant soap on Anopheles gambiae s.s (Diptera: Culicidae) in laboratory and semifield environs.

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Mdoe, France P; Nkwengulila, Gamba; Chobu, Mariam; Lyaruu, Lucile; Gyunda, Israel L; Mbepera, Saada; Xue, Rui-De; Kweka, Eliningaya J

2014-05-03

Mosquito larval control using chemicals and biological agents is of paramount importance in vector population and disease incidence reduction. A commercial synthetic disinfectant soap was evaluated against larvae of Anopheles gambiae s.s. in both laboratory and semi field conditions. Five concentrations of commercial synthetic disinfectant soap (0.0001, 0.001, 0.01, 0.1 and 1%) were prepared and evaluated against third instar larvae in laboratory and semi field environments. Mortality was scored at 12, 24, 48, and 72 hrs. Each dosage had 6 replicates, having twenty 3rd instar larvae of An.gambiae s.s. In the laboratory phase, all dosages had significantly higher larval mortalities than in controls, while in semi field conditions, the dosages of 0.0001, 0.001 and 0.01% had lower mortalities than laboratory trials. In the comparison between semi field and laboratory trials, only 0.1 and 1% dosage had significant difference with more mortality in semifield conditions. Proportions of larvae that died during mortality monitoring intervals in laboratory and semi field had significant differences only at 12 hrs and 72 hrs. The findings of this study have demonstrated that the mortality of larvae caused by commercial synthetic disinfectant soap is worth further studies in open water bodies. More studies are necessary to find out the effect of sunlight on the chemistry of the synthetic disinfectant and other variables in small scale full field trials.

Biological activities of four essential oils against Anopheles gambiae ...

African Journals Online (AJOL)

The control of malaria is still a challenge partly due to mosquito's resistance to current available insecticides. The aim of this work was to evaluate the ovicidal, larvicidal and repellent activities of Lantana camara, Hyptis suaveolens, Hyptis spicigera and Ocimum canum essential oils against Anopheles gambiae s.l. ...

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

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Bayoh Nabie M

2007-02-01

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

Susceptibility of Anopheles gambiae to insecticides used for malaria vector control in Rwanda.

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Hakizimana, Emmanuel; Karema, Corine; Munyakanage, Dunia; Iranzi, Gad; Githure, John; Tongren, Jon Eric; Takken, Willem; Binagwaho, Agnes; Koenraadt, Constantianus J M

2016-12-01

The widespread emergence of resistance to pyrethroids is a major threat to the gains made in malaria control. To monitor the presence and possible emergence of resistance against a variety of insecticides used for malaria control in Rwanda, nationwide insecticide resistance surveys were conducted in 2011 and 2013. Larvae of Anopheles gambiae sensu lato mosquitoes were collected in 12 sentinel sites throughout Rwanda. These were reared to adults and analysed for knock-down and mortality using WHO insecticide test papers with standard diagnostic doses of the recommended insecticides. A sub-sample of tested specimens was analysed for the presence of knockdown resistance (kdr) mutations. A total of 14,311 mosquitoes were tested and from a sample of 1406 specimens, 1165 (82.9%) were identified as Anopheles arabiensis and 241 (17.1%) as Anopheles gambiae sensu stricto. Mortality results indicated a significant increase in resistance to lambda-cyhalothrin from 2011 to 2013 in 83% of the sites, permethrin in 25% of the sites, deltamethrin in 25% of the sites and DDT in 50% of the sites. Mosquitoes from 83% of the sites showed full susceptibility to bendiocarb and 17% of sites were suspected to harbour resistance that requires further confirmation. No resistance was observed to fenitrothion in all study sites during the entire survey. The kdr genotype results in An. gambiae s.s. showed that 67 (50%) possessed susceptibility (SS) alleles, while 35 (26.1%) and 32 (23.9%) mosquitoes had heterozygous (RS) and homozygous (RR) alleles, respectively. Of the 591 An. arabiensis genotyped, 425 (71.9%) possessed homozygous (SS) alleles while 158 (26.7%) and 8 (1.4%) had heterozygous (RS) and homozygous (RR) alleles, respectively. Metabolic resistance involving oxidase enzymes was also detected using the synergist PBO. This is the first nationwide study of insecticide resistance in malaria vectors in Rwanda. It shows the gradual increase of insecticide resistance to pyrethroids (lambda

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

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Nwane, Philippe; Etang, Josiane; Chouaїbou, Mouhamadou; Toto, Jean Claude; Koffi, Alphonsine; Mimpfoundi, Rémy; Simard, Frédéric

2013-02-22

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

Efficacy of bifenthrin-impregnated bednets against Anopheles funestus and pyrethroid-resistant Anopheles gambiae in North Cameroon

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Chouaibou Mouhamadou

2006-09-01

Full Text Available Abstract Background Recent field studies indicated that insecticide-treated bednets (ITNs maintain their efficacy despite a high frequency of the knock-down resistance (kdr gene in Anopheles gambiae populations. It was essential to evaluate ITNs efficacy in areas with metabolic-based resistance. Methods Bifenthrin was used in this experiment because it is considered a promising candidate for bednets impregnation. Nets were treated at 50 mg/m2, a dose that has high insecticidal activity on kdr mosquitoes and at 5 mg/m2, a dose that kills 95% of susceptible mosquitoes under laboratory conditions with 3 minutes exposure. Bednets were holed to mimic physical damage. The trial was conducted in three experimental huts from Pitoa, North-Cameroon where Anopheles gambiae displays metabolic resistance and cohabits with An. funestus. Results Bifenthrin at 50 mg/m2 significantly reduced anophelines' entry rate (>80%. This was not observed at 5 mg/m2. Both treatments increased exophily in An. gambiae, and to a lesser extent in An. funestus. With bifenthrin at high dosage, over 60% reduction in blood feeding and 75–90% mortality rates were observed in both vectors. Despite presence of holes, only a single An. gambiae and two An. funestus females were collected inside the treated net, and all were found dead. The same trends were observed with low dosage bifenthrin though in most cases, no significant difference was found with the untreated control net. Conclusion Bifenthrin-impregnated bednets at 50 mg/m2 were efficient in the reduction of human-vector contact in Pitoa. Considerable personal protection was gained against An. funestus and metabolic pyrethroid resistant An. gambiae populations.

Efficacy of bifenthrin-impregnated bednets against Anopheles funestus and pyrethroid-resistant Anopheles gambiae in North Cameroon

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Chouaibou, Mouhamadou; Simard, Frédéric; Chandre, Fabrice; Etang, Josiane; Darriet, Frédéric; Hougard, Jean-Marc

2006-01-01

Background Recent field studies indicated that insecticide-treated bednets (ITNs) maintain their efficacy despite a high frequency of the knock-down resistance (kdr) gene in Anopheles gambiae populations. It was essential to evaluate ITNs efficacy in areas with metabolic-based resistance. Methods Bifenthrin was used in this experiment because it is considered a promising candidate for bednets impregnation. Nets were treated at 50 mg/m2, a dose that has high insecticidal activity on kdr mosquitoes and at 5 mg/m2, a dose that kills 95% of susceptible mosquitoes under laboratory conditions with 3 minutes exposure. Bednets were holed to mimic physical damage. The trial was conducted in three experimental huts from Pitoa, North-Cameroon where Anopheles gambiae displays metabolic resistance and cohabits with An. funestus. Results Bifenthrin at 50 mg/m2 significantly reduced anophelines' entry rate (>80%). This was not observed at 5 mg/m2. Both treatments increased exophily in An. gambiae, and to a lesser extent in An. funestus. With bifenthrin at high dosage, over 60% reduction in blood feeding and 75–90% mortality rates were observed in both vectors. Despite presence of holes, only a single An. gambiae and two An. funestus females were collected inside the treated net, and all were found dead. The same trends were observed with low dosage bifenthrin though in most cases, no significant difference was found with the untreated control net. Conclusion Bifenthrin-impregnated bednets at 50 mg/m2 were efficient in the reduction of human-vector contact in Pitoa. Considerable personal protection was gained against An. funestus and metabolic pyrethroid resistant An. gambiae populations. PMID:16961938

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

NARCIS (Netherlands)

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

2011-01-01

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

Characterizing the insecticide resistance of Anopheles gambiae in Mali.

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Cisse, Moussa B M; Keita, Chitan; Dicko, Abdourhamane; Dengela, Dereje; Coleman, Jane; Lucas, Bradford; Mihigo, Jules; Sadou, Aboubacar; Belemvire, Allison; George, Kristen; Fornadel, Christen; Beach, Raymond

2015-08-22

The impact of indoor residual spraying (IRS) and long-lasting insecticide nets (LLINs), key components of the national malaria control strategy of Mali, is threatened by vector insecticide resistance. The objective of this study was to assess the level of insecticide resistance in Anopheles gambiae sensu lato populations from Mali against four classes of insecticide recommended for IRS: organochlorines (OCs), pyrethroids (PYs), carbamates (CAs) and organophosphates (OPs). Characterization of resistance was done in 13 sites across southern Mali and assessed presence and distribution of physiological mechanisms that included target-site modifications: knockdown resistance (kdr) and altered acetycholinesterase (AChE), and/or metabolic mechanisms: elevated esterases, glutathione S-transferases (GSTs), and monooxygenases. The World Health Organization (WHO) tube test was used to determine phenotypic resistance of An. gambiae s.l. to: dichlorodiphenyltrichloroethane (DDT) (OC), deltamethrin (PY), lambda-cyhalothrin (PY), bendiocarb (CA), and fenitrothion (OP). Identification of sibling species and presence of the ace-1 (R) and Leu-Phe kdr, resistance-associated mutations, were determined using polymerase chain reaction (PCR) technology. Biochemical assays were conducted to detect increased activity of GSTs, oxidases and esterases. Populations tested showed high levels of resistance to DDT in all 13 sites, as well as increased resistance to deltamethrin and lambda-cyhalothrin in 12 out of 13 sites. Resistance to fenitrothion and bendiocarb was detected in 1 and 4 out of 13 sites, respectively. Anopheles coluzzii, An. gambiae sensu stricto and Anopheles arabiensis were identified with high allelic frequencies of kdr in all sites where each of the species were found (13, 12 and 10 sites, respectively). Relatively low allelic frequencies of ace-1 (R) were detected in four sites where this assessment was conducted. Evidence of elevated insecticide metabolism, based on oxidase

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.

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Dziedzom K de Souza

Full Text Available 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.

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

DEFF Research Database (Denmark)

Belmont, Martin; Cazzamali, Giuseppe; Williamson, Michael

2006-01-01

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

G protein-coupled receptors in Anopheles gambiae.

Science.gov (United States)

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

2002-10-04

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.

Insecticide resistance status of three malaria vectors, Anopheles gambiae (s.l.), An. funestus and An. mascarensis, from the south, central and east coasts of Madagascar.

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Rakotoson, Jean-Desire; Fornadel, Christen M; Belemvire, Allison; Norris, Laura C; George, Kristen; Caranci, Angela; Lucas, Bradford; Dengela, Dereje

2017-08-23

Insecticide-based vector control, which comprises use of insecticide-treated bed nets (ITNs) and indoor residual spraying (IRS), is the key method to malaria control in Madagascar. However, its effectiveness is threatened as vectors become resistant to insecticides. This study investigated the resistance status of malaria vectors in Madagascar to various insecticides recommended for use in ITNs and/or IRS. WHO tube and CDC bottle bioassays were performed on populations of Anopheles gambiae (s.l.), An. funestus and An. mascarensis. Adult female An. gambiae (s.l.) mosquitoes reared from field-collected larvae and pupae were tested for their resistance to DDT, permethrin, deltamethrin, alpha-cypermethrin, lambda-cyhalothrin, bendiocarb and pirimiphos-methyl. Resting An. funestus and An. mascarensis female mosquitoes collected from unsprayed surfaces were tested against permethrin, deltamethrin and pirimiphos-methyl. The effect on insecticide resistance of pre-exposure to the synergists piperonyl-butoxide (PBO) and S,S,S-tributyl phosphorotrithioate (DEF) also was assessed. Molecular analyses were done to identify species and determine the presence of knock-down resistance (kdr) and acetylcholinesterase resistance (ace-1 R ) gene mutations. Anopheles funestus and An. mascarensis were fully susceptible to permethrin, deltamethrin and pirimiphos-methyl. Anopheles gambiae (s.l.) was fully susceptible to bendiocarb and pirimiphos-methyl. Among the 17 An. gambiae (s.l.) populations tested for deltamethrin, no confirmed resistance was recorded, but suspected resistance was observed in two sites. Anopheles gambiae (s.l.) was resistant to permethrin in four out of 18 sites (mortality 68-89%) and to alpha-cypermethrin (89% mortality) and lambda-cyhalothrin (80% and 85%) in one of 17 sites, using one or both assay methods. Pre-exposure to PBO restored full susceptibility to all pyrethroids tested except in one site where only partial restoration to permethrin was observed. DEF

Larvicidal activity of six Nigerian plant species against Anopheles ...

African Journals Online (AJOL)

This study evaluated the larvicidal activity of extracts from six Nigerian plant species (Zanthoxylum zanthoxyloides, Piper guineense, Nicotianat abacum, Erythrophleum suaveoleus, Jatropha curcas and Petiveria alliacea) against laboratory-bred Anopheles gambiae and Aedes aegypti larvae. Zanthoxylum zanthoxyloides ...

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

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Antonio-Nkondjio, Christophe; Youmsi-Goupeyou, Marlene; Kopya, Edmond; Tene-Fossog, Billy; Njiokou, Flobert; Costantini, Carlo; Awono-Ambene, Parfait

2014-08-03

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. 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. 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 (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. 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.

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

NARCIS (Netherlands)

Takken, W.; Lindsay, S.W.

2003-01-01

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

Insertion polymorphisms of SINE200 retrotransposons within speciation islands of Anopheles gambiae molecular forms

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Tu Zhijian

2008-08-01

Full Text Available Abstract Background SINEs (Short INterspersed Elements are homoplasy-free and co-dominant genetic markers which are considered to represent useful tools for population genetic studies, and could help clarifying the speciation processes ongoing within the major malaria vector in Africa, Anopheles gambiae s.s. Here, we report the results of the analysis of the insertion polymorphism of a nearly 200 bp-long SINE (SINE200 within genome areas of high differentiation (i.e. "speciation islands" of M and S A. gambiae molecular forms. Methods A SINE-PCR approach was carried out on thirteen SINE200 insertions in M and S females collected along the whole range of distribution of A. gambiae s.s. in sub-Saharan Africa. Ten specimens each for Anopheles arabiensis, Anopheles melas, Anopheles quadriannulatus A and 15 M/S hybrids from laboratory crosses were also analysed. Results Eight loci were successfully amplified and were found to be specific for A. gambiae s.s.: 5 on 2L chromosome and one on X chromosome resulted monomorphic, while two loci positioned respectively on 2R (i.e. S200 2R12D and X (i.e. S200 X6.1 chromosomes were found to be polymorphic. S200 2R12D was homozygote for the insertion in most S-form samples, while intermediate levels of polymorphism were shown in M-form, resulting in an overall high degree of genetic differentiation between molecular forms (Fst = 0.46 p S200 X6.1 was found to be fixed in all M- and absent in all S-specimens. This led to develop a novel easy-to-use PCR approach to straightforwardly identify A. gambiae molecular forms. This novel approach allows to overcome the constraints associated with markers on the rDNA region commonly used for M and S identification. In fact, it is based on a single copy and irreversible SINE200 insertion and, thus, is not subjected to peculiar evolutionary patterns affecting rDNA markers, e.g. incomplete homogenization of the arrays through concerted evolution and/or mixtures of M and S IGS

Genome mapping and characterization of the Anopheles gambiae heterochromatin

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Sharakhova Maria V

2010-08-01

Full Text Available Abstract Background Heterochromatin plays an important role in chromosome function and gene regulation. Despite the availability of polytene chromosomes and genome sequence, the heterochromatin of the major malaria vector Anopheles gambiae has not been mapped and characterized. Results To determine the extent of heterochromatin within the An. gambiae genome, genes were physically mapped to the euchromatin-heterochromatin transition zone of polytene chromosomes. The study found that a minimum of 232 genes reside in 16.6 Mb of mapped heterochromatin. Gene ontology analysis revealed that heterochromatin is enriched in genes with DNA-binding and regulatory activities. Immunostaining of the An. gambiae chromosomes with antibodies against Drosophila melanogaster heterochromatin protein 1 (HP1 and the nuclear envelope protein lamin Dm0 identified the major invariable sites of the proteins' localization in all regions of pericentric heterochromatin, diffuse intercalary heterochromatin, and euchromatic region 9C of the 2R arm, but not in the compact intercalary heterochromatin. To better understand the molecular differences among chromatin types, novel Bayesian statistical models were developed to analyze genome features. The study found that heterochromatin and euchromatin differ in gene density and the coverage of retroelements and segmental duplications. The pericentric heterochromatin had the highest coverage of retroelements and tandem repeats, while intercalary heterochromatin was enriched with segmental duplications. We also provide evidence that the diffuse intercalary heterochromatin has a higher coverage of DNA transposable elements, minisatellites, and satellites than does the compact intercalary heterochromatin. The investigation of 42-Mb assembly of unmapped genomic scaffolds showed that it has molecular characteristics similar to cytologically mapped heterochromatin. Conclusions Our results demonstrate that Anopheles polytene chromosomes

The role of skin microbiota in the attractiveness of humans to the malaria mosquito Anopheles gambiae Giles

NARCIS (Netherlands)

Verhulst, N.O.

2010-01-01

Malaria is one of the most serious infectious diseases in the world. The African mosquito Anopheles gambiae sensu stricto (henceforth termed An. gambiae) is highly competent for malaria parasites and preferably feeds on humans inside houses, which make it one of the most effective vectors of the

Human skin microbiota and their volatiles as odour baits for the malaria mosquito Anopheles gambiae s.s

NARCIS (Netherlands)

Verhulst, N.O.; Mukabana, W.R.; Takken, W.; Smallegange, R.C.

2011-01-01

Host seeking by the malaria mosquito Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) is mainly guided by volatile chemicals present in human odours. The skin microbiota plays an important role in the production of these volatiles, and skin bacteria grown on agar plates attract An. gambiae

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

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Le Goff Gilbert

2003-10-01

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

Daily oviposition patterns of the African malaria mosquito Anopheles gambiae Giles (Diptera: Culicidae on different types of aqueous substrates

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Knols Bart GJ

2004-12-01

Full Text Available Abstract Background Anopheles gambiae Giles is the most important vector of human malaria in sub-Saharan Africa. Knowledge of the factors that influence its daily oviposition pattern is crucial if field interventions targeting gravid females are to be successful. This laboratory study investigated the effect of oviposition substrate and time of blood feeding on daily oviposition patterns of An. gambiae mosquitoes. Methods Greenhouse-reared gravid and hypergravid (delayed oviposition onset An. gambiae sensu stricto and wild-caught An. gambiae sensu lato were exposed to three types of substrates in choice and no-choice cage bioassays: water from a predominantly anopheline colonised ground pool (anopheline habitat water, swamp water mainly colonised by culicine larvae (culicine habitat water and distilled water. The daily oviposition pattern and the number of eggs oviposited on each substrate during the entire egg-laying period were determined. The results were subjected to analysis of variance using the General Linear Model (GLM procedure. Results The main oviposition time for greenhouse-reared An. gambiae s.s. was between 19:00 and 20:00 hrs, approximately one hour after sunset. Wild-caught gravid An. gambiae s.l. displayed two distinct peak oviposition times between 19:00 and 20:00 hrs and between 22:00 and 23:00 hrs, respectively. During these times, both greenhouse-reared and wild-caught mosquitoes significantly (P P Conclusion This study shows that the peak oviposition time of An. gambiae s.l. may be regulated by the light-dark cycle rather than oviposition habitat characteristics or feeding times. However, the number of eggs laid by the female mosquito during the peak oviposition time is affected by the suitability of the habitat.

The effects of oviposition-site deprivation on Anopheles gambiae reproduction

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Dieter Kathryne L

2012-10-01

Full Text Available Abstract Background The African malaria mosquito, Anopheles gambiae, depends on availability of suitable surface water for oviposition. Short and long dry spells occur throughout the year in many parts of its range that limit its access to oviposition sites. Although not well understood, oviposition-site deprivation has been found to rapidly reduce egg batch size and hatch rate of several mosquito species. We conducted laboratory experiments to assess these effects of oviposition-site deprivation on An. gambiae and to evaluate the role of nutrition and sperm viability as mediators of these effects. Methods Anopheles gambiae adults (1–2 d old from the G3 laboratory colony were assigned to the following treatment groups: oviposition-deprived (fed once and then deprived of oviposition site for 7 or 14 d, multiple-fed control (fed regularly once a week and allowed to lay eggs without delay, and age matched blood-deprived control (fed once, three days before water for oviposition was provided. Egg batch size and hatch rate were measured. In the second experiment two additional treatment groups were included: oviposition-deprived females that received either a second (supplemental blood meal or virgin males (supplemental mating 4 days prior to receiving water for oviposition. Results An. gambiae was highly sensitive to oviposition-site deprivation. Egg batch size dropped sharply to 0–3.5 egg/female within 14 days, due to reduced oviposition rate rather than a reduced number of eggs/batch. Egg hatch rate also fell dramatically to 0-2% within 7 days. The frequency of brown eggs that fail to tan was elevated. A supplemental blood meal, but not ‘supplemental insemination,’ recovered the oviposition rate of females subjected to oviposition-site deprivation. Similarly, a supplemental blood meal, but not ‘supplemental insemination,’ partly recovered hatch rate, but this increase was marginally significant (P  Conclusions Even a short dry spell

Larvicidal effect of aqueous and ethanolic extracts of Senna alata on Anopheles gambiae, Culex quinquefasciatus and Aedes aegypti.

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Edwin, Ubulom Peace Mayen; Nyiutaha, Imandeh Godwin; Essien, Akpabio Eno; Nnamdi, Opara Kenneth; Sunday, Ekanem Mfon

2013-05-01

Senna alata is locally used in South Eastern Nigeria in the treatment of several infections which include ringworm and other parasitic skin diseases.The larvicidal activities of aqueous and ethanolic leaf and stem extracts of S. alata were evaluated in static bioassays, on fourth instar larvae of Anopheles gambiae, Culex quinquefasciatus and Aedes aegypti, at extract concentrations of 0.15, 0.30, 0.45, 0.60 and 0.75% w/v, for 72 hours. Mortality of larvae exposed to the different extracts increased with increase in extract concentration and time of exposure. This study revealed a differential potency of the extracts used and a difference in susceptibility of larvae to the extracts as evident by the 72hLC₅₀ values obtained. The leaf extract proved to be more lethal to the larvae than the stem extract as judged by the 72hLC₅₀ values obtained both for the aqueous as well as the ethanolic extracts assayed. Phytochemical screening of the plant parts investigated revealed the presence of some plant metabolites, which have been reported in separate studies to be lethal to mosquito larvae. Results obtained from this study suggest that the leaf and stem extracts of S. alata possess a promising larvicidal potential which can be exploited in mosquito vector control.

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

DEFF Research Database (Denmark)

Schöller, Susanne; Belmont, Martin; Cazzamali, Giuseppe

2005-01-01

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

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

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Kamareddine Layla

2013-01-01

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

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

NARCIS (Netherlands)

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

2004-01-01

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

Spatial repellency screening in a high-throughput apparatus with Aedes aegypti and Anopheles gambiae

Science.gov (United States)

Spatial repellents are essential for personal protection against mosquitoes, such as Aedes aegypti and Anopheles gambiae, to reduce annoyance biting and transmission of mosquito-borne diseases. The number of safe and effective repellents, including DEET, picaridin, and IR3535, is limited and contin...

Long non-coding RNA discovery across the genus anopheles reveals conserved secondary structures within and beyond the Gambiae complex.

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Jenkins, Adam M; Waterhouse, Robert M; Muskavitch, Marc A T

2015-04-23

Long non-coding RNAs (lncRNAs) have been defined as mRNA-like transcripts longer than 200 nucleotides that lack significant protein-coding potential, and many of them constitute scaffolds for ribonucleoprotein complexes with critical roles in epigenetic regulation. Various lncRNAs have been implicated in the modulation of chromatin structure, transcriptional and post-transcriptional gene regulation, and regulation of genomic stability in mammals, Caenorhabditis elegans, and Drosophila melanogaster. The purpose of this study is to identify the lncRNA landscape in the malaria vector An. gambiae and assess the evolutionary conservation of lncRNAs and their secondary structures across the Anopheles genus. Using deep RNA sequencing of multiple Anopheles gambiae life stages, we have identified 2,949 lncRNAs and more than 300 previously unannotated putative protein-coding genes. The lncRNAs exhibit differential expression profiles across life stages and adult genders. We find that across the genus Anopheles, lncRNAs display much lower sequence conservation than protein-coding genes. Additionally, we find that lncRNA secondary structure is highly conserved within the Gambiae complex, but diverges rapidly across the rest of the genus Anopheles. This study offers one of the first lncRNA secondary structure analyses in vector insects. Our description of lncRNAs in An. gambiae offers the most comprehensive genome-wide insights to date into lncRNAs in this vector mosquito, and defines a set of potential targets for the development of vector-based interventions that may further curb the human malaria burden in disease-endemic countries.

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

NARCIS (Netherlands)

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

2009-01-01

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

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

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Etang Josiane

2008-09-01

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

Evidence of multiple insecticide resistance mechanisms in Anopheles gambiae populations in Bangui, Central African Republic.

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Olé Sangba, Marina Lidwine; Sidick, Aboubakar; Govoetchan, Renaud; Dide-Agossou, Christian; Ossè, Razaki A; Akogbeto, Martin; Ndiath, Mamadou Ousmane

2017-01-13

Knowledge of insecticide resistance status in the main malaria vectors is an essential component of effective malaria vector control. This study presents the first evaluation of the status of insecticide resistance in Anopheles gambiae populations from Bangui, the Central African Republic. Anopheles mosquitoes were reared from larvae collected in seven districts of Bangui between September to November 2014. The World Health Organisation's bioassay susceptibility tests to lambda-cyhalothrin (0.05%), deltamethrin (0.05%), DDT (4%), malathion (5%), fenitrothion (1%) and bendiocarb (0.1%) were performed on adult females. Species and molecular forms as well as the presence of L1014F kdr and Ace-1 R mutations were assessed by PCR. Additional tests were conducted to assess metabolic resistance status. After 1 h exposure, a significant difference of knockdown effect was observed between districts in all insecticides tested except deltamethrin and malathion. The mortality rate (MR) of pyrethroids group ranging from 27% (CI: 19-37.5) in Petevo to 86% (CI: 77.6-92.1) in Gbanikola; while for DDT, MR ranged from 5% (CI: 1.6-11.3) in Centre-ville to 39% (CI: 29.4-49.3) in Ouango. For the organophosphate group a MR of 100% was observed in all districts except Gbanikola where a MR of 96% (CI: 90-98.9) was recorded. The mortality induced by bendiocarb was very heterogeneous, ranging from 75% (CI: 62.8-82.8) in Yapele to 99% (CI: 84.5-100) in Centre-ville. A high level of kdr-w (L1014F) frequency was observed in all districts ranging from 93 to 100%; however, no kdr-e (L1014S) and Ace-1 R mutation were found in all tested mosquitoes. Data of biochemical analysis showed significant overexpression activities of cytochrome P450, GST and esterases in Gbanikola and Yapele (χ 2  = 31.85, df = 2, P resistance to DDT and pyrethroids as well as precocious emergence of resistance to carbamates were detected among A. gambiae mosquitoes from Bangui, including target-site mutations

Analysis of near infrared spectra for age-grading of wild populations of Anopheles gambiae.

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Krajacich, Benjamin J; Meyers, Jacob I; Alout, Haoues; Dabiré, Roch K; Dowell, Floyd E; Foy, Brian D

2017-11-07

Understanding the age-structure of mosquito populations, especially malaria vectors such as Anopheles gambiae, is important for assessing the risk of infectious mosquitoes, and how vector control interventions may impact this risk. The use of near-infrared spectroscopy (NIRS) for age-grading has been demonstrated previously on laboratory and semi-field mosquitoes, but to date has not been utilized on wild-caught mosquitoes whose age is externally validated via parity status or parasite infection stage. In this study, we developed regression and classification models using NIRS on datasets of wild An. gambiae (s.l.) reared from larvae collected from the field in Burkina Faso, and two laboratory strains. We compared the accuracy of these models for predicting the ages of wild-caught mosquitoes that had been scored for their parity status as well as for positivity for Plasmodium sporozoites. Regression models utilizing variable selection increased predictive accuracy over the more common full-spectrum partial least squares (PLS) approach for cross-validation of the datasets, validation, and independent test sets. Models produced from datasets that included the greatest range of mosquito samples (i.e. different sampling locations and times) had the highest predictive accuracy on independent testing sets, though overall accuracy on these samples was low. For classification, we found that intramodel accuracy ranged between 73.5-97.0% for grouping of mosquitoes into "early" and "late" age classes, with the highest prediction accuracy found in laboratory colonized mosquitoes. However, this accuracy was decreased on test sets, with the highest classification of an independent set of wild-caught larvae reared to set ages being 69.6%. Variation in NIRS data, likely from dietary, genetic, and other factors limits the accuracy of this technique with wild-caught mosquitoes. Alternative algorithms may help improve prediction accuracy, but care should be taken to either maximize

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

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Nkya, Theresia E; Akhouayri, Idir; Poupardin, Rodolphe; Batengana, Bernard; Mosha, Franklin; Magesa, Stephen; Kisinza, William; David, Jean-Philippe

2014-01-25

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. 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. 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 strain and supported the impact

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

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Kayode David Ileke

2015-07-01

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

The Seasonality and Ecology of the Anopheles gambiae complex (Dipetra: Culicidae) in Liberia Using Molecular Identification.

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Fahmy, N T; Villinski, J T; Bolay, F; Stoops, C A; Tageldin, R A; Fakoli, L; Okasha, O; Obenauer, P J; Diclaro, J W

2015-05-01

Members of the Anopheles gambiae sensu lato (Giles) complex define a group of seven morphologically indistinguishable species, including the principal malaria vectors in Sub-Saharan Africa. Members of this complex differ in behavior and ability to transmit malaria; hence, precise identification of member species is critical to monitoring and evaluating malaria threat levels. We collected mosquitoes from five counties in Liberia every other month from May 2011 until May 2012, using various trapping techniques. A. gambiae complex members were identified using molecular techniques based on differences in the ribosomal DNA (rDNA) region between species and the molecular forms (S and M) of A. gambiae sensu stricto (s.s) specimens. In total, 1,696 A. gambiae mosquitoes were collected and identified. DNA was extracted from legs of each specimen with species identification determined by multiplex polymerase chain reaction using specific primers. The molecular forms (M or S) of A. gambiae s.s were determined by restriction fragment length polymorphism. Bivariate and multivariate logistic regression models identified environmental variables associated with genomic differentiation. Our results indicate widespread occurrence of A. gambiae s.s., the principal malaria vector in the complex, although two Anopheles melas Theobald/A. merus Donitz mosquitoes were detected. We found 72.6, 25.5, and 1.9% of A. gambiae s.s specimens were S, M, and hybrid forms, respectively. Statistical analysis indicates that the S form was more likely to be found in rural areas during rainy seasons and indoor catchments. This information will enhance vector control efforts in Liberia. Published by Oxford University Press on behalf of Entomological Society of America 2015. This work is written by US Government employees and is in the public domain in the US.

Costs of insensitive acetylcholinesterase insecticide resistance for the malaria vector Anopheles gambiae homozygous for the G119S mutation

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Noel Valérie

2010-01-01

Full Text Available Abstract Background The G119S mutation responsible for insensitive acetylcholinesterase resistance to organophosphate and carbamate insecticides has recently been reported from natural populations of Anopheles gambiae in West Africa. These reports suggest there are costs of resistance associated with this mutation for An. gambiae, especially for homozygous individuals, and these costs could be influential in determining the frequency of carbamate resistance in these populations. Methods Life-history traits of the AcerKis and Kisumu strains of An. gambiae were compared following the manipulation of larval food availability in three separate experiments conducted in an insecticide-free laboratory environment. These two strains share the same genetic background, but differ in being homozygous for the presence or absence of the G119S mutation at the ace-1 locus, respectively. Results Pupae of the resistant strain were significantly more likely to die during pupation than those of the susceptible strain. Ages at pupation were significantly earlier for the resistant strain and their dry starved weights were significantly lighter; this difference in weight remained when the two strains were matched for ages at pupation. Conclusions The main cost of resistance found for An. gambiae mosquitoes homozygous for the G119S mutation was that they were significantly more likely to die during pupation than their susceptible counterparts, and they did so across a range of larval food conditions. Comparing the frequency of G119S in fourth instar larvae and adults emerging from the same populations would provide a way to test whether this cost of resistance is being expressed in natural populations of An. gambiae and influencing the dynamics of this resistance mutation.

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

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Gouagna Louis C

2007-04-01

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

Malaria transmission and insecticide resistance of Anopheles gambiae in Libreville and Port-Gentil, Gabon

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Kombila Maryvonne

2010-11-01

Full Text Available Abstract Background Urban malaria is a major health priority for civilian and militaries populations. A preliminary entomologic study has been conducted in 2006-2007, in the French military camps of the two mains towns of Gabon: Libreville and Port-Gentil. The aim was to assess the malaria transmission risk for troops. Methods Mosquitoes sampled by human landing collection were identified morphologically and by molecular methods. The Plasmodium falciparum circumsporozoïte (CSP indexes were measured by ELISA, and the entomological inoculation rates (EIR were calculated for both areas. Molecular assessments of pyrethroid knock down (kdr resistance and of insensitive acetylcholinesterase resistance were conducted. Results In Libreville, Anopheles gambiae s.s. S form was the only specie of the An. gambiae complex present and was responsible of 9.4 bites per person per night. The circumsporozoïte index was 0.15% and the entomological inoculation rate estimated to be 1.23 infective bites during the four months period. In Port-Gentil, Anopheles melas (75.5% of catches and An. gambiae s.s. S form (24.5% were responsible of 58.7 bites per person per night. The CSP indexes were of 1.67% for An. gambiae s.s and 0.28% for An. melas and the EIRs were respectively of 1.8 infective bites per week and of 0.8 infective bites per week. Both kdr-w and kdr-e mutations in An. gambiae S form were found in Libreville and in Port-Gentil. Insensitive acetylcholinesterase has been detected for the first time in Gabon in Libreville. Conclusion Malaria transmission exists in both town, but with high difference in the level of risk. The co-occurrence of molecular resistances to the main families of insecticide has implications for the effectiveness of the current vector control programmes that are based on pyrethroid-impregnated bed nets.

Impact of Diurnal Temperature Fluctuations during Larval Development on Adult Life History Traits and Insecticide Susceptibility in Two Vectors; Anopheles gambiae and Aedes aegypti

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2014-04-30

ANOPHELES GAMBIAE AND AEDES AEGYPTI. by Jeffrey W. Clark Dissertation submitted to the Faculty of the Department of Preventive Medicine and...Vectors; Anopheles gambiae and Aedes aegypti." Name of Candidate: Jeffrey Clark Doctor of Philosophy Degree April 30, 2014 DISSERTATION AND ABSTRACT...for the many fruitful discussions and the standing offer to help whenever I needed it; and to Joe Wagman, for providing needed Aedes aegypti eggs from

Anopheles gambiae antiviral immune response to systemic O'nyong-nyong infection.

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Joanna Waldock

Full Text Available 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.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.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 antiviral immune response in A. gambiae is thus

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

NARCIS (Netherlands)

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

2007-01-01

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

Larval competition between An. coluzzii and An. gambiae in insectary and semi-field conditions in Burkina Faso.

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Gimonneau, Geoffrey; Brossette, Lou; Mamaï, Wadaka; Dabiré, Roch K; Simard, Frédéric

2014-02-01

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, Pcompetition 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 processes using inbred laboratory colonies in

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

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Teklu, B. M.; Tekie, H.; McCartney, M.; Kibret, S.

2010-08-01

Entomological studies to determine the effect of the physical characteristics of larval breeding water bodies and reservoir water level changes on the occurrence of Anopheles mosquito larvae and on the spatial and temporal formation of larval breeding habitats were conducted in two villages at Koka reservoir between August and December 2007. Of the two study villages, Ejersa is in close proximity to the reservoir, and Kuma is 5 km away from it. Data on the type, number and physical characteristics of Anopheles larval breeding habitat, species composition and densities of anopheles mosquitoes in and around the study villages were investigated and recorded. Meteorological and reservoir water level data were compared with availability of Anopheles larval breeding sites and densities. Entomological data from the weekly larval collections showed that Anopheles pharoensis Theobald, Anopheles gambiae s.l. Giles, Anopheles coustani Laveran and Anopheles squamosus Theobald were breeding in the study area. The mean larval density of A. gambiae s.l. in this study was higher in slightly turbid and shallow aquatic habitats than in turbid and relatively deep aquatic habitats (F=16.97, plevels and the number of positive breeding habitats at Ejersa during the sampling period (r=0.605, pphysical characteristics such as water temperature, turbidity, depth and vegetation cover play an important role in the species composition, total Anopheles larval count, and the density of Anopheles mosquitoes in the vicinity. The proliferation of suitable breeding habitats around the reservoir villages is strongly associated with reservoir water level changes. This is particularly important for A. pharoensis and A. arabiensis which are important vectors of malaria in the area. Further investigation on the species diversity, physical and chemical habitat characteristics and impact of water holding capacity of the soil need to be done to generate detailed baseline data which will serve as a basis

Effects of fungal infection on feeding and survival of Anopheles gambiae (Diptera: Culicidae) on plant sugars

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Ondiaka, S.N.; Masinde, E.W.; Koenraadt, C.J.M.; Takken, W.; Mukabana, W.R.

2015-01-01

Background The entomopathogenic fungus Metarhizium anisopliae shows great promise for the control of adult malaria vectors. A promising strategy for infection of mosquitoes is supplying the fungus at plant feeding sites. Methods We evaluated the survival of fungus-exposed Anopheles gambiae

Ultrastructure of a microsporidium brachiola gambiae n.sp.parasitising a mosquito anopheles gamblae, a malaria vector

Czech Academy of Sciences Publication Activity Database

Weiser, Jaroslav; Žižka, Zdeněk

- (2003), s. 35-36 ISSN 1214-021X. [Conference on Cell Biology /5./. České Budějovice, 08.09.2003-10.09.2003] Institutional research plan: CEZ:AV0Z5020903 Keywords : anopheles gambiae * malaria * vector Subject RIV: EE - Microbiology, Virology

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

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Tchoumbougnang F.

2009-01-01

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

Larvicidal potential of some plants from West Africa against Culex quinquefasciatus (Say) and Anopheles gambiae Giles (Diptera: Culicidae).

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Azokou, Alain; Koné, Mamidou W; Koudou, Benjamin G; Tra Bi, Honora F

2013-01-01

Mosquitoes increased resistance to insecticides, and environmental concerns about the use of insecticides, pose a major challenge in the search for new molecules to deplete and incapacitate mosquito populations. Plants are the valuable source as practices consisting in exploiting plant materials as repellents, and are still in wide use throughout developing countries. The aim of the present study was to screen plants from Cτte d'Ivoire for larvicidal activity against mosquitoes. Resistant and sensitive larvae (III and IV instar) of Anopheles gambiae and Culex quinquefasciatus were exposed to crude ethanol extracts (90%) of 45 plants and viability observed after 30 min, 6, 12 and 24 h postincubation. After partition of active extracts, each fraction (hexane and chloroform washed with NaCl 1%, tannins and aqueous) was tested using the same protocol at various concentrations (1000- 31.2 ppm). Of 49 extracts tested, 7 exhibited high potential (LC50 = 80 to 370 ppm) against resistant and sensitive III and IV instar larvae of An. gambiae and Cx. quinquefasciatus. These extracts were from Cissus populnea, Cochlospermum planchonii, Heliotropium indicum, Phyllanthus amarus, Vitex grandifolia and Alchornea cordifolia. However, three most active plant species (LC50 = 80- 180 ppm) were Cs. populnea, Cm. planchonii and P. amarus Their hexane and chloroform fractions showed high larvicidal activity. This study demonstrated that plants from Cτte d'Ivoire have a real potential for malaria, yellow fever, filarial and dengue vector control. Those could be used as sources or provide lead compounds for the development of safe plant-based biocides.

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

NARCIS (Netherlands)

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

2010-01-01

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

Analysis of near infrared spectra for age-grading of wild populations of Anopheles gambiae

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Benjamin J. Krajacich

2017-11-01

Full Text Available Abstract Background Understanding the age-structure of mosquito populations, especially malaria vectors such as Anopheles gambiae, is important for assessing the risk of infectious mosquitoes, and how vector control interventions may impact this risk. The use of near-infrared spectroscopy (NIRS for age-grading has been demonstrated previously on laboratory and semi-field mosquitoes, but to date has not been utilized on wild-caught mosquitoes whose age is externally validated via parity status or parasite infection stage. In this study, we developed regression and classification models using NIRS on datasets of wild An. gambiae (s.l. reared from larvae collected from the field in Burkina Faso, and two laboratory strains. We compared the accuracy of these models for predicting the ages of wild-caught mosquitoes that had been scored for their parity status as well as for positivity for Plasmodium sporozoites. Results Regression models utilizing variable selection increased predictive accuracy over the more common full-spectrum partial least squares (PLS approach for cross-validation of the datasets, validation, and independent test sets. Models produced from datasets that included the greatest range of mosquito samples (i.e. different sampling locations and times had the highest predictive accuracy on independent testing sets, though overall accuracy on these samples was low. For classification, we found that intramodel accuracy ranged between 73.5–97.0% for grouping of mosquitoes into “early” and “late” age classes, with the highest prediction accuracy found in laboratory colonized mosquitoes. However, this accuracy was decreased on test sets, with the highest classification of an independent set of wild-caught larvae reared to set ages being 69.6%. Conclusions Variation in NIRS data, likely from dietary, genetic, and other factors limits the accuracy of this technique with wild-caught mosquitoes. Alternative algorithms may help improve

Biological differences in reproductive strategy between the mosquito sibling species Anopheles gambiae sensu stricto and An. quadriannulatus

NARCIS (Netherlands)

Takken, W.; Stuke, K.; Klowden, M.J.

2002-01-01

Females of the afrotropical mosquito species Anopheles gambiae Giles sensu stricto and An. quadriannulatus (Theobald) (Diptera: Culicidae) were studied for the effect of blood meal size and the frequency of blood feeding on reproductive development during the first gonotrophic cycle. To standardize

Factors influencing the spatial distribution of Anopheles larvae in Coimbatore District, Tamil Nadu, India.

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Arjunan, Naresh Kumar; Kadarkarai, Murugan; Kumar, Shobana; Pari, Madhiyazhagan; Thiyagarajan, Nataraj; Vincent, C Thomas; Barnard, Donald R

2015-12-01

Malaria causes extensive morbidity and mortality in humans and results in significant economic losses in India. The distribution of immature malaria-transmitting Anopheles mosquitoes was studied in 17 villages in Coimbatore District as a prelude to the development and implementation of vector control strategies that are intended to reduce the risk of human exposure to potentially infectious mosquitoes. Eight Anopheles species were recorded. The most numerous species were Anopheles vagus, Anopheles subpictus, and Anopheles hyrcanus. The location of mosquito development sites and the density of larvae in each village was evaluated for correlation with selected demographic, biologic, and land use parameters using remote sensing and geographic information systems (GIS) technology. We found the number of mosquito development sites in a village and the density of larvae in such sites to be positively correlated with human population density but not the surface area (km(2)) of the village. The number of mosquito development sites and the density of larvae in each site were not correlated. Data from this study are being used to construct a GIS-based mapping system that will enable the location of aquatic habitats with Anopheles larvae in the Coimbatore District, Tamil Nadu, India as target sites for the application of vector control. Copyright © 2015 Elsevier B.V. All rights reserved.

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

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Sisay Dugassa

Full Text Available 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.Experiments were implemented in an 80 m(2 semi-field system where 200 gravid Anopheles gambiae s.s. were released nightly. The efficacy of the Box, CDC and Frommer updraft gravid traps was compared. The Box gravid trap was tested to determine if the presence of the trap over water and the trap's sound affected catch size. Mosquitoes approaching the treatment were evaluated using electrocuting nets or detergents added to the water in the trap. Based on the results, a new gravid trap (OviART trap that provided an open, unobstructed oviposition site was developed and evaluated.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.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.

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

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Mahairaki Vassiliki

2007-01-01

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

Effects of larvicidal and larval nutritional stresses on Anopheles gambiae development, survival and competence for Plasmodium falciparum.

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Vantaux, Amélie; Ouattarra, Issiaka; Lefèvre, Thierry; Dabiré, Kounbobr Roch

2016-04-23

Many studies have shown that the environment in which larvae develop can influence adult characteristics with consequences for the transmission of pathogens. We investigated how two environmental stresses (larviciding and nutritional stress) interact to affect Anopheles gambiae (previously An. gambiae S molecular form) life history traits and its susceptibility for field isolates of its natural malaria agent Plasmodium falciparum. Larvae were reared in the presence or not of a sub-lethal concentration of larvicide and under a high and low food regimen. Development time, individual size, adult survival and competence for P. falciparum were assessed. Individuals under low food regimen took more time to develop, had a lower development success and were smaller while there was no main effect of larvicide exposure on these traits. However, larvicide exposure impacted individual size in interaction with nutritional stress. Female survival was affected by the interaction between gametocytemia, parasite exposure and larval diet, as well as the interaction between gametocytemia, parasite exposure and larvicidal stress, and the interaction between gametocytemia, larvicidal exposure and larval diet. Among the 951 females dissected 7 days post-infection, 559 (58.78%) harboured parasites. Parasite prevalence was significantly affected by the interaction between larvicidal stress and larval diet. Indeed, females under low food regimen had a higher prevalence than females under high food regimen and this difference was greater under larvicidal stress. The two stresses did not impact parasite intensity. We found that larval nutritional and larvicidal stresses affect mosquito life history traits in complex ways, which could greatly affect P. falciparum transmission. Further studies combining field-based trials on larvicide use and mosquito experimental infections would give a more accurate understanding of the effects of this vector control tool on malaria transmission.

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

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

2002-01-01

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

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

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Dominique C. K. Sohounhloué

2010-12-01

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

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

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Petrarca Vincenzo

2011-07-01

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

Mediation of oviposition site selection in the African malaria mosquito Anopheles gambiae (Diptera: Culicidae) by semiochemicals of microbial origin

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Sumba, L.A.; Guda, T.O.; Deng, A.L.; Hassanali, A.; Beier, J.C.; Knols, B.G.J.

2004-01-01

Laboratory studies were carried out to investigate the role of larval habitat-derived microorganisms in the production of semiochemicals for oviposition site selection by Anopheles gambiae Giles sensu stricto mosquitoes. Dual-choice bioassays with gravid females were conducted in standard mosquito

Evaluation of active ingredients and larvicidal activity of clove and cinnamon essential oils against Anopheles gambiae (sensu lato).

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Thomas, Adelina; Mazigo, Humphrey D; Manjurano, Alphaxard; Morona, Domenica; Kweka, Eliningaya J

2017-09-06

Mosquitoes are well-known vectors of many diseases including malaria and lymphatic filariasis. Uses of synthetic insecticides are associated with high toxicity, resistance, environmental pollution and limited alternative, effective synthetic insecticides. This study was undertaken to evaluate the larvicidal efficacy of clove and cinnamon essential oils against laboratory Anopheles gambiae (sensu stricto) and wild An. arabiensis larvae. The standard WHO guideline for larvicides evaluation was used, and the GC-MS machine was used for active compounds percentage composition analysis and structures identification. Probit regression analysis was used for LC 50 and LC 95 calculations while a t-test was used to test for significant differences between laboratory-reared and wild larvae populations in each concentration of plant extract. Mortality effect of clove and cinnamon essential oils against wild and laboratory-reared larvae had variations indicated by their LC 50 and LC 95 values. The mortality at different concentrations of cinnamon and clove post-exposure for wild and laboratory-reared larvae were dosage-dependent and were higher for cinnamon than for clove essential oils. The mortality effect following exposure to a blend of the two essential oils was higher for blends containing a greater proportion of cinnamon oil. In the chemical analysis of the active ingredients of cinnamon essential oil, the main chemical content was Eugenol, and the rarest was β-Linalool while for clove essential oil, the main chemical content was Eugenol and the rarest was Bicyclo. The essential oils showed a larvicidal effect which was concentration-dependent for both laboratory and wild collected larvae. The active ingredient compositions triggered different responses in mortality. Further research in small-scale should be conducted with concentrated extracted compounds.

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

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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.

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

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

2009-01-01

Background A fast development of urban agriculture has recently taken place in many areas in the Republic of Benin. This study aims to assess the rapid expansion of urban agriculture especially, its contribution to the emergence of insecticide resistance in populations of Anopheles gambiae. Methods The protocol was based on the collection of sociological data by interviewing vegetable farmers regarding various agricultural practices and the types of pesticides used. Bioassay tests were performed to assess the susceptibility of malaria vectors to various agricultural insecticides and biochemical analysis were done to characterize molecular status of population of An. gambiae. Results This research showed that: (1) The rapid development of urban agriculture is related to unemployment observed in cities, rural exodus and the search for a balanced diet by urban populations; (2) Urban agriculture increases the farmers' household income and their living standard; (3) At a molecular level, PCR revealed the presence of three sub-species of An. gambiae (An. gambiae s.s., Anopheles melas and Anopheles arabiensis) and two molecular forms (M and S). The kdr west mutation recorded in samples from the three sites and more specifically on the M forms seems to be one of the major resistance mechanisms found in An. gambiae from agricultural areas. Insecticide susceptibility tests conducted during this research revealed a clear pattern of resistance to permethrin (76% mortality rate at Parakou; 23.5% at Porto-Novo and 17% at Cotonou). Conclusion This study confirmed an increase activity of the vegetable farming in urban areas of Benin. This has led to the use of insecticide in an improper manner to control vegetable pests, thus exerting a huge selection pressure on mosquito larval population, which resulted to the emergence of insecticide resistance in malaria vectors. PMID:19442297

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

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Rodrigues, J.; Oliveira, G. A.; Kotsyfakis, Michalis; Dixit, R.; Molina-Cruz, A.; Jochim, R.; Barillas-Mury, C.

2012-01-01

Roč. 7, č. 4 (2012), e35210 E-ISSN 1932-6203 Institutional support: RVO:60077344 Keywords : malaria * mosquito * serine protease * sporozoites * ookinetes * gene silencing * midgut * salivary glands * Plasmodium falciparum * Anopheles gambiae Subject RIV: EB - Genetics ; Molecular Biology Impact factor: 3.730, year: 2012 http://www.plosone.org/article/info%3Adoi%2F10.1371%2Fjournal.pone.0035210

A reliable morphological method to assess the age of male Anopheles gambiae

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Killeen Gerry F

2006-07-01

Full Text Available Abstract Background Release of genetically-modified (GM or sterile male mosquitoes for malaria control is hampered by inability to assess the age and mating history of free-living male Anopheles. Methods Age and mating-related changes in the reproductive system of male Anopheles gambiae were quantified and used to fit predictive statistical models. These models, based on numbers of spermatocysts, relative size of sperm reservoir and presence/absence of a clear area around the accessory gland, were evaluated using an independent sample of mosquitoes whose status was blinded during the experiment. Results The number of spermatocysts in male testes decreased with age, and the relative size of their sperm reservoir increased. The presence of a clear area around accessory glands was also linked to age and mating status. A quantitative model was able to categorize males from the blind trial into age groups of young (≤ 4 days and old (> 4 days with an overall efficiency of 89%. Using the parameters of this model, a simple table was compiled that can be used to predict male age. In contrast, mating history could not be reliably assessed as virgins could not be distinguished from mated males. Conclusion Simple assessment of a few morphological traits which are easily collected in the field allows accurate age-grading of male An. gambiae. This simple, yet robust, model enables evaluation of demographic patterns and mortality in wild and released males in populations targeted by GM or sterile male-based control programmes.

Laboratory evaluation of the irritancy of bendiocarb, lambda-cyhalothrin and DDT to Anopheles gambiae.

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Evans, R G

1993-09-01

In a laboratory study, the irritancy of bendiocarb, lambda-cyhalothrin and DDT to Anopheles gambiae was evaluated at field, 1/3 field and 1/10 field rates using WHO conical exposure chambers and excito-repellency test boxes. Bendiocarb was the least irritant insecticide at all rates, inducing levels of takeoff, flight and exiting behavior similar to those of a distilled water control treatment. Of those mosquitoes introduced to the bendiocarb-treated boxes, not more than 1% exited and survived at any dose rate. Lambda-cyhalothrin and DDT were highly irritant to An. gambiae, inducing a strong stimulation to take off and fly and also a high level of exiting. Exiting-survival rates associated with lambda-cyhalothrin and DDT were between 15 and 51%. The relevance of these findings to the control of mosquito populations and the prevention of malaria transmission is discussed.

Deforestation and Vectorial Capacity of Anopheles gambiae Giles Mosquitoes in Malaria Transmission, Kenya

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Afrane, Yaw A.; Little, Tom J.; Lawson, Bernard W.; Githeko, Andrew K.

2008-01-01

We investigated the effects of deforestation on microclimates and sporogonic development of Plasmodium falciparum parasites in Anopheles gambiae mosquitoes in an area of the western Kenyan highland prone to malaria epidemics. An. gambiae mosquitoes were fed with P. falciparum–infected blood through membrane feeders. Fed mosquitoes were placed in houses in forested and deforested areas in a highland area (1,500 m above sea level) and monitored for parasite development. Deforested sites had higher temperatures and relative humidities, and the overall infection rate of mosquitoes was increased compared with that in forested sites. Sporozoites appeared on average 1.1 days earlier in deforested areas. Vectorial capacity was estimated to be 77.7% higher in the deforested site than in the forested site. We showed that deforestation changes microclimates, leading to more rapid sporogonic development of P. falciparum and to a marked increase of malaria risk in the western Kenyan highland. PMID:18826815

Chloroquine mediated modulation of Anopheles gambiae gene expression.

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Patrícia Abrantes

2008-07-01

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

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

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Céline Christiansen-Jucht

2015-05-01

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

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

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Derua Yahya A

2012-06-01

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

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

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Aristide Sawdetuo Hien

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

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

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Oduola Adedayo O

2012-06-01

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

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

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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.

Laboratory evolution of the entomopathogenic fungus Beauveria bassiana against Anopheles stephensi larvae (Diptera: Culicidae

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Rahele Veys-Behbahani

2014-09-01

Full Text Available Objective: To examine Beauveria bassiana (B. bassiana fungus bioassay against the larval stages of Anopheles stephensi in Iran. Methods: The fungal suspension by the concentrations of 1伊1 09, 5伊1 08, 1 08, 5伊1 07 and 1伊 107 conidia per milliliter have been prepared in different volumes (2, 4 and 6 mL and each concentration were added to containers containing 25 Anopheles larva instars 1 and 2. The mortality of the dead larvae with abnormal symptoms was recorded as a result of the fungal infection after 24, 48 and 72 h. Results: Comparison between the mean mortality rate of Anopheles stephensi larva at different concentrations of B. bassiana strain Iran 429C at 2, 4 and 6 mL showed that there was no significant relation of the mean mortality rate of larvae at concentrations of 1伊109 and 5伊108, and after 48 h resulted in 100% mortality rate of the larvae populations. In addition, there is no significant differences in the amounts of lethal times (LT (LT50 and LT90 as LT90 values calculated at a concentration of 5伊108 and in volumes 2, 4 and 6 mL were 1.46, 1.36 and 1.08 d, respectively. Conclusions: B. bassiana strain Iran 429C in 2 mL of 5伊108 concentration or the concentration of a 1伊109 mL per 100 mL of water is recommended as the optimal concentration for the control of Anopheles larvae. The development of suitable formulations of entomopathogenic fungi may be a promising prospect in the mosquito control programs.

Spatial distribution of the chromosomal forms of anopheles gambiae in Mali

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Traoré Sékou F

2008-10-01

Full Text Available Abstract Background Maps of the distribution of malaria vectors are useful tools for stratification of malaria risk and for selective vector control strategies. Although the distribution of members of the Anopheles gambiae complex is well documented in Africa, a continuous map of the spatial distribution of the chromosomal forms of An. gambiae s.s. is not yet available at country level to support control efforts. Methods Bayesian geostatistical methods were used to produce continuous maps of the spatial distribution of the chromosomal forms of An. gambiae s.s. (Mopti, Bamako, Savanna and their hybrids/recombinants based on their relative frequencies in relation to climatic and environmental factors in Mali. Results The maps clearly show that each chromosomal form favours a particular defined eco-climatic zone. The Mopti form prefers the dryer northern Savanna and Sahel and the flooded/irrigated areas of the inner delta of the Niger River. The Savanna form favours the Sudan savanna areas, particularly the South and South-Eastern parts of the country (Kayes and Sikasso regions. The Bamako form has a strong preference for specific environmental conditions and it is confined to the Sudan savanna areas around urban Bamako and the Western part of Sikasso region. The hybrids/recombinants favour the Western part of the country (Kayes region bordering the Republic of Guinea Conakry. Conclusion The maps provide valuable information for selective vector control in Mali (insecticide resistance management and may serve as a decision support tool for the basis for future malaria control strategies including genetically manipulated mosquitoes.

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

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N. Sogoba

2007-05-01

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

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

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

2016-11-19

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

Enkapsulasi B. bassiana menggunakan maizena dan daya infeksinya terhadap larva Aedes aegypti, Anopheles sp., Culex sp.

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Mutiara Widawati

2015-12-01

Full Text Available Abstract. Encapsulation formulae of mycoinsecticide have to be able to maintain fungus viability and pathogenicity. This mycoinsecticide was developed as an alternative way to control mosquito borne disease. The aim of this study was to encapsulate Beauveria bassiana as viable storage and have the capability to kill larvae of Aedes aegypti, Anopheles sp. and Culex sp. Mosquito larvae obtained from laboratory reared at health research laboratory; Loka Litbang P2B2 Ciamis. The treatments made in this study were the formu-lation of cornstarch and controls for comparison. This study showed potential formulation of cornstarch encapsulation as a biolarvacidal. Cornstarch formulations proven to be succeed in maintaining fungus viability, however, the pathogenicity of the microcapsule still not effective to kill Aedes, Culex and Anopheles larvae. Keywords: encapsulation, Beauveria bassiana, Ae.aegypti, Anopheles sp., Culex sp. Abstrak. Pembuatan formula bioinsektisida yang optimal sebagai salah satu alternatif untuk pengen-dalian nyamuk vektor perlu di kembangkan. Sediaan mikoinsektisida yang dibuat harus dapat memper-tahankan viabilitas jamur B. bassiana sehingga masih efektif pada saat penggunaannya. Salah satu cara yang digunakan untuk menjaga kestabilan sediaan mikoinsektisida yang berdampak langsung pada via-bilitas jamur adalah dengan menerapkan metode enkapsulasi. Penelitian ini bertujuan untuk meng-hasilkan sediaan mikokapsul dari Beauvaria bassiana melalui proses enkapsulasi menggunakan maizena yang memiliki kapabilitas tinggi sebagai penyimpan B. bassiana dan efektif dalam membunuh larva dan telur Aedes aegypti, Anopheles sp. dan Culex sp. Semua larva uji berasal dari insektarium laboratorium penelitian kesehatan Loka litbang P2B2 Ciamis. Pembuatan enkapsulasi dimulai dengan kultur dan pema-nenan B. bassiana, uji viabilitas, proses enkapsulasi serta uji larvasida di laboratorium. Uji dilakukan dengan satu perlakuan dan satu kontrol untuk

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

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Caccone Adalgisa

2008-04-01

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

A Simple Key for Identifying the Sibling Species of the Malaria Vector Anopheles gambiae (Giles Complex by Polytene Chromosome Cytogenetics

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Music Temitope OBEMBE

2018-03-01

Full Text Available It has been established that Anopheles gambiae complex sibling species are the major Plasmodium malaria vectors in Africa; however, not all the sibling species transmit the infection. Easier molecular methods, PCR-based assays, have been developed to distinguish the several members of the A. gambiae complex. However, malaria vector research in less developed countries, particularly sub-Saharan Africa, is being hampered by the lack of PCR facilities in laboratories and the cost of carrying out the assay within lack of funding. Hence, the present study was designed to develop a simple identification key, based on an affordable method of polytene chromosome cytotaxonomy, for identifying the major P. falciparum vectors. The Identification Key was successfully used to identify two members of the A. gambiae complex, A. gambiae sensu stricto and A. arabiensis, which are the most potent malaria vectors in Africa; even so, it could not be used to establish the infective and the refractory strains.

Sex-specific and blood meal-induced proteins of Anopheles gambiae midguts: analysis by two-dimensional gel electrophoresis

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Laurent-Winter C

2003-02-01

Full Text Available Abstract Background Anopheles gambiae is the main vector of Plasmodium falciparum in Africa. The mosquito midgut constitutes a barrier that the parasite must cross if it is to develop and be transmitted. Despite the central role of the mosquito midgut in the host/parasite interaction, little is known about its protein composition. Characterisation of An. gambiae midgut proteins may identify the proteins that render An. gambiae receptive to the malaria parasite. Methods We carried out two-dimensional gel electrophoresis of An. gambiae midgut proteins and compared protein profiles for midguts from males, sugar-fed females and females fed on human blood. Results Very few differences were detected between male and female mosquitoes for the approximately 375 silver-stained proteins. Male midguts contained ten proteins not detected in sugar-fed or blood-fed females, which are therefore probably involved in male-specific functions; conversely, female midguts contained twenty-three proteins absent from male midguts. Eight of these proteins were specific to sugar-fed females, and another ten, to blood-fed females. Conclusion Mass spectrometry analysis of the proteins found only in blood-fed female midguts, together with data from the recent sequencing of the An. gambiae genome, should make it possible to determine the role of these proteins in blood digestion or parasite receptivity.

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

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Badolo Athanase

2012-07-01

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

Efficacy of mosquito nets treated with a pyrethroid-organophosphorous mixture against Kdr- and Kdr+ malaria vectors (Anopheles gambiae

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Darriet F.

2003-12-01

Full Text Available In order to prevent the resistance of Anopheles gambiae s.l. to pyrethroids from spreading too quickly and to lengthen the effectiveness of insecticide impregnated mosquito nets, it has recently been suggested to use mixtures of insecticides that have different modes of action. This study presents the results obtained with tulle mosquito nets treated with bifenthrin (a pyrethroid] and chlorpyrifos-methyl (an organophosphorous both separately and in mixture on two strains of An. gambiae, one sensitive to all insecticides, and the other resistant to pyrethroids. The values of KDt50 and KDt95 and the mortality induced with the mixture of bifenthrin (25 mg/m2 and chlorpyrifos-methyl (4.5 mg/m2 show a significant synergistic effect on the strain of An. gambiae susceptible to insecticides. However, the tested combination does not induce any synergistic effect on the VKPR strain selected with permethrin, but only enhances the effectiveness of the two insecticides taken separately.

The response of the malaria mosquito, Anopheles gambiae, to two components of human sweat, ammonia and L-lactic acid, in an olfactometer

NARCIS (Netherlands)

Braks, M.A.H.; Meijerink, J.; Takken, W.

2001-01-01

In an olfactometer study on the response of the anthropophilic malaria mosquito Anopheles gambiae s.s. (Diptera, Culicidae) to human sweat it was found that freshly collected sweat, mostly of eccrine origin, was attractive, but that incubated sweat was significantly more attractive than fresh sweat.

Characterisation of Anopheles gambiae heme oxygenase and metalloporphyrin feeding suggests a potential role in reproduction.

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Spencer, Christopher S; Yunta, Cristina; de Lima, Glauber Pacelli Gomes; Hemmings, Kay; Lian, Lu-Yun; Lycett, Gareth; Paine, Mark J I

2018-05-03

The mosquito Anopheles gambiae is the principal vector for malaria in sub-Saharan Africa. The ability of A. gambiae to transmit malaria is strictly related to blood feeding and digestion, which releases nutrients for oogenesis, as well as substantial amounts of highly toxic free heme. Heme degradation by heme oxygenase (HO) is a common protective mechanism, and a gene for HO exists in the An. gambiae genome HO (AgHO), although it has yet to be functionally examined. Here, we have cloned and expressed An. gambiae HO (AgHO) in E. coli. Purified recombinant AgHO bound hemin stoichiometrically to form a hemin-enzyme complex similar to other HOs, with a K D of 3.9 ± 0.6 μM; comparable to mammalian and bacterial HOs, but 7-fold lower than that of Drosophila melanogaster HO. AgHO also degraded hemin to biliverdin and released CO and iron in the presence of NADPH cytochrome P450 oxidoreductase (CPR). Optimal AgHO activity was observed at 27.5 °C and pH 7.5. To investigate effects of AgHO inhibition, adult female A. gambiae were fed heme analogues Sn- and Zn-protoporphyrins (SnPP and ZnPP), known to inhibit HO. These led to a dose dependent decrease in oviposition. Cu-protoporphyrin (CuPP), which does not inhibit HO had no effect. These results demonstrate that AgHO is a catalytically active HO and that it may play a key role in egg production in mosquitoes. It also presents a potential target for the development of compounds aimed at sterilising mosquitoes for vector control. Copyright © 2018 Elsevier Ltd. All rights reserved.

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

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Chilaka Nora

2012-01-01

Full Text Available Abstract 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.

Field evaluation of spatial repellency of metofluthrin-impregnated plastic strips against Anopheles gambiae complex in Bagamoyo, coastal Tanzania.

OpenAIRE

Kawada, Hitoshi; Temu, Emmanuel A.; Minjas, Japhet N.; Matsumoto, Osamu; Iwasaki, Tomonori; Takagi, Masahiro

2008-01-01

Metofluthrin is a newly synthesized pyrethroid possessing high knockdown and lethal activity against mosquitoes. Studies of metofluthrin-impregnated plastic strips have been performed with dengue vectors. This study reports the efficacy of the new prototypes of metofluthrin-impregnated plastic strips against malaria vectors, Anopheles gambiae complex, in the Kongo villages of Bagamoyo district in coastal Tanzania. The study, using 20 houses, half intervention, half control, was conducted for ...

Seasonal climate effects anemotaxis in newly emerged adult Anopheles gambiae Giles in Mali, West Africa.

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Manoukis, Nicholas C; Baber, Ibrahima; Diallo, Moussa; Sogoba, Nafomon; Ribeiro, José M C

2011-01-01

The direction and magnitude of movement by 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 the existence of attractants in the vicinity. We report here the direction of movement of newly emerged An. gambiae in nature, around the village of Donéguébougou, Mali. We assessed the direction of movement for individual mosquitoes by placing them in a novel enclosure with exit traps oriented in the direction of the cardinal and intermediate points of the compass. We consistently found predominantly Southward directions of movement during 2009 and 2010, with an additional Eastward component during the dry season and a Westward one during the wet season. Our data indicate that wind has an important effect on the direction of movement, but that this effect varied by season: Average directions of movement were downwind during the dry season and upwind during the wet season. A switch in anemotactic response suggests that the direction of movement of An. gambiae relative to the wind immediately after emergence under varying conditions of humidity should be further investigated under controlled conditions.

Seasonal climate effects anemotaxis in newly emerged adult Anopheles gambiae Giles in Mali, West Africa.

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Nicholas C Manoukis

Full Text Available The direction and magnitude of movement by 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 the existence of attractants in the vicinity. We report here the direction of movement of newly emerged An. gambiae in nature, around the village of Donéguébougou, Mali. We assessed the direction of movement for individual mosquitoes by placing them in a novel enclosure with exit traps oriented in the direction of the cardinal and intermediate points of the compass. We consistently found predominantly Southward directions of movement during 2009 and 2010, with an additional Eastward component during the dry season and a Westward one during the wet season. Our data indicate that wind has an important effect on the direction of movement, but that this effect varied by season: Average directions of movement were downwind during the dry season and upwind during the wet season. A switch in anemotactic response suggests that the direction of movement of An. gambiae relative to the wind immediately after emergence under varying conditions of humidity should be further investigated under controlled conditions.

Transcription profiling of a recently colonised pyrethroid resistant Anopheles gambiae strain from Ghana

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Donnelly Martin J

2007-01-01

Full Text Available Abstract Background Mosquito resistance to the pyrethroid insecticides used to treat bednets threatens the sustainability of malaria control in sub-Saharan Africa. While the impact of target site insensitivity alleles is being widely discussed the implications of insecticide detoxification – though equally important – remains elusive. The successful development of new tools for malaria intervention and management requires a comprehensive understanding of insecticide resistance, including metabolic resistance mechanisms. Although three enzyme families (cytochrome P450s, glutathione S-transferases and carboxylesterases have been widely associated with insecticide detoxification the role of individual enzymes is largely unknown. Results Here, constitutive expression patterns of genes putatively involved in conferring pyrethroid resistance was investigated in a recently colonised pyrethroid resistant Anopheles gambiae strain from Odumasy, Southern Ghana. RNA from the resistant strain and a standard laboratory susceptible strain, of both sexes was extracted, reverse transcribed and labelled with either Cy3- or Cy5-dye. Labelled cDNA was co-hybridised to the detox chip, a custom-made microarray containing over 230 A. gambiae gene fragments predominantly from enzyme families associated with insecticide resistance. After hybridisation, Cy3- and Cy5-signal intensities were measured and compared gene by gene. In both females and males of the resistant strain the cytochrome P450s CYP6Z2 and CYP6M2 are highly over-expressed along with a member of the superoxide dismutase (SOD gene family. Conclusion These genes differ from those found up-regulated in East African strains of pyrethroid resistant A. gambiae and constitute a novel set of candidate genes implicated in insecticide detoxification. These data suggest that metabolic resistance may have multiple origins in A. gambiae, which has strong implications for the management of resistance.

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

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

2014-01-01

Objective Insecticide resistance molecular markers can provide sensitive indicators of resistance development in Anopheles vector populations. Assaying these makers is of paramount importance in the resistance monitoring programme. We investigated the presence and distribution of knock-down resistance (kdr) mutations in Anopheles gambiae s.l. in Tanzania. Methods Indoor-resting Anopheles mosquitoes were collected from 10 sites and tested for insecticide resistance using the standard WHO protocol. Polymerase chain reaction-based molecular diagnostics were used to genotype mosquitoes and detect kdr mutations. Results The An. gambiae tested were resistance to lambdacyhalothrin in Muheza, Arumeru and Muleba. Out of 350 An. gambiae s.l. genotyped, 35% were An. gambiae s.s. and 65% An. arabiensis. L1014S and L1014F mutations were detected in both An. gambiae s.s. and An. arabiensis. L1014S point mutation was found at the allelic frequency of 4–33%, while L1014F was at the allelic frequency 6–41%. The L1014S mutation was much associated with An. gambiae s.s. (χ2 = 23.41; P protocolo estándar de la OMS. Mediante un diagnóstico molecular basado en la PCR se genotiparon los mosquitos y se detectaron los genotipos kdr. Resultados Los An. gambiae evaluados eran resistentes a lambdacialotrina en Muheza, Arumeru y Muleba. De 350 An. gambiae s.l. genotipados, 35% eran An. gambiae s.s. y 65% eran An. arabiensis. Se detectaron mutaciones L1014S y L1014F tanto en An. gambiae s.s. como en An. arabiensis. La mutación puntual L1014S se encontró con una frecuencia alélica de 4-33%, mientras que L1014F tenía una frecuencia alélica de 6-14%. La mutación L1014S estaba ampliamente asociada a An. gambiae s.s. (Chi-Cuadrado = 23.41; P < 0.0001) y la L1014F estaba asociada con An. arabiensis (Chi-Square = 11.21; P = 0.0008). El alelo L1014S estaba significativamente asociado con mosquitos resistentes a la lambdacialotrina (P < 0.001). Conclusión La

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

NARCIS (Netherlands)

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

2013-01-01

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

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.

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Mbare, Oscar; Lindsay, Steven W; Fillinger, Ulrike

2013-03-14

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. 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. 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% of eggs laid by females exposed to 0

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

Science.gov (United States)

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

The bionomics of Anopheles merus (Diptera: Culicidae along the Kenyan coast

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Kipyab Pamela C

2013-02-01

Full Text Available Abstract Background Anopheles merus, a sibling species of the Anopheles gambiae complex occurs along the East African coast but its biology and role in malaria transmission in this region is poorly understood. We evaluated the blood feeding pattern and the role of this species in malaria transmission in Malindi district, Coastal Kenya. Methods Adult mosquitoes were collected indoors by CDC light traps and Pyrethrum Spray Catch and outdoors by CDC light traps. Anopheles females were identified to species by morphological characteristics and sibling species of An. gambiae complex distinguished by rDNA polymerase chain reaction (PCR. Screening for host blood meal sources and presence or absence of Plasmodium falciparum circumsporozoite proteins was achieved by Enzyme Linked Immunosorbent Assays (ELISA. Results Anopheles merus comprised 77.8% of the 387 Anopheles gambiae s.l adults that were collected. Other sibling species of Anopheles gambiae s.l identified in the study site included An. arabiensis(3.6%, and An. gambiae s.s. (8%. The human blood index for An. merus was 0.12, while the sporozoite rate was 0.3%. Conclusion These findings suggest that An. merus can play a minor role in malaria transmission along the Kenyan Coast and should be a target for vector control which in turn could be applied in designing and implementing mosquito control programmes targeting marsh-breeding mosquitoes; with the ultimate goal being to reduce the transmission of malaria associated with these vectors.

Vector competence of Anopheles and Culex mosquitoes for Zika virus

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Brittany L. Dodson

2017-03-01

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

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

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Verhulst, Niels O; Mbadi, Phoebe A; Kiss, Gabriella Bukovinszkiné; Mukabana, Wolfgang R; van Loon, Joop J A; Takken, Willem; Smallegange, Renate C

2011-02-08

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

Susceptibility of female Anopheles mosquito to pyrethroid ...

African Journals Online (AJOL)

The detection of insecticides resistance status in a natural population of Anopheles vectors is a vital tool for malaria control intervention strategy against Anopheles gambiae sensu lato, which is the main malaria vector in Nigeria. This study was conducted to determine the susceptibility status of the female Anopheles ...

Additional selection for insecticide resistance in urban malaria vectors: DDT resistance in Anopheles arabiensis from Bobo-Dioulasso, Burkina Faso.

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Christopher M Jones

Full Text Available In the city of Bobo-Dioulasso in Burkina Faso, Anopheles arabiensis has superseded Anopheles gambiae s.s. as the major malaria vector and the larvae are found in highly polluted habitats normally considered unsuitable for Anopheles mosquitoes. Here we show that An. gambiae s.l. adults emerging from a highly polluted site in the city centre (Dioulassoba have a high prevalence of DDT resistance (percentage mortality after exposure to diagnostic dose=65.8% in the dry season and 70.4% in the rainy season, respectively. An investigation into the mechanisms responsible found an unexpectedly high frequency of the 1014S kdr mutation (allele frequency=0.4, which is found at very low frequencies in An. arabiensis in the surrounding rural areas, and an increase in transcript levels of several detoxification genes, notably from the glutathione transferase and cytochrome P450 gene families. A number of ABC transporter genes were also expressed at elevated levels in the DDT resistant An. arabiensis. Unplanned urbanisation provides numerous breeding grounds for mosquitoes. The finding that Anopheles mosquitoes adapted to these urban breeding sites have a high prevalence of insecticide resistance has important implications for our understanding of the selective forces responsible for the rapid spread of insecticide resistant populations of malaria vectors in Africa.

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

NARCIS (Netherlands)

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

2008-01-01

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

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

Science.gov (United States)

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.

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

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Rian, Sigrid Katrine Eivindsdatter

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

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

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Lyimo Issa N

2009-12-01

Full Text Available Abstract The entomopathogenic fungus Metarhizium anisopliae isolate IP 46, originating from a soil sample collected in 2001 in the Cerrado of Central Brazil, was tested for its ability to reduce the survival of adult male and female Anopheles gambiae s.s. and An. arabiensis mosquitoes. A 6-h exposure to the fungus coated on test paper at a concentration of 3.3 × 106 conidia cm-2 reduced the daily survival of both mosquito species (HR = 3.14, p An. gambiae s.s relative to An. arabiensis (HR = 1.38, p 95% of mosquito cadavers in the treatment groups. The results indicate that M. anisopliae IP 46 has the potential to be a bio-control agent for African malaria vector species, and is a suitable candidate for further research and development.

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

OpenAIRE

Kaufmann, Christian; Betschart, Bruno

2007-01-01

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

Riceland mosquito management practices for Anopheles quadrimaculatus larvae.

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Allen, R A; Wilkes, W W; Lewis, C N; Meisch, M V

2008-12-01

Two separate but related studies were conducted regarding management of Anopheles quadrimaculatus larval populations in commercial rice fields near Cleveland, MS, in 2004. Study 1 was to evaluate the effectiveness of 2 treatments of aerially applied ultra-low volume applications of Bacillus thuringiensis var. israelensis (Bti) against An. quadrimaculatus larvae in dense, high-canopy mid- to late-season rice crop. Study 2 was to investigate the effect of preflood treatments of lambda-cyhalothrin (Karate), which is commonly used against rice water weevil (Lissorhoptrus oryzophilus), on An. quadrimaculatus larvae. Excellent initial, but short residual control (>99% control 1 day after treatment) was observed in the Bti-treated fields in both mid- and late-season rice. Little or no effect on mosquito larvae was observed in the lambda-cyhalothrin-treated fields. Results indicate that Bti can be effectively used by mosquito management personnel to control larval populations of An. quadrimaculatus in late-season rice fields; however, lambda-cyhalothrin did not effectively control larval An. quadrimaculatus when applied preflood to rice fields.

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

NARCIS (Netherlands)

Foster, W.A.; Takken, W.

2004-01-01

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°C both sexes generally die if

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

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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. PMID:24675797

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

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Kalle Magnusson

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

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

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Lorenz Lena M

2011-12-01

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

The larvicidal effects of black pepper (Piper nigrum L.) and piperine against insecticide resistant and susceptible strains of Anopheles malaria vector mosquitoes.

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Samuel, Michael; Oliver, Shüné V; Coetzee, Maureen; Brooke, Basil D

2016-04-26

Insecticide resistance carries the potential to undermine the efficacy of insecticide based malaria vector control strategies. Therefore, there is an urgent need for new insecticidal compounds. Black pepper (dried fruit from the vine, Piper nigrum), used as a food additive and spice, and its principal alkaloid piperine, have previously been shown to have larvicidal properties. The aim of this study was to investigate the larvicidal effects of ground black pepper and piperine against third and fourth instar Anopheles larvae drawn from several laboratory-reared insecticide resistant and susceptible strains of Anopheles arabiensis, An. coluzzii, An. gambiae, An. quadriannulatus and An. funestus. Larvae were fed with mixtures of standard larval food and either ground black pepper or piperine in different proportions. Mortality was recorded 24 h after black pepper and 48 h after piperine were applied to the larval bowls. Black pepper and piperine mixtures caused high mortality in the An. gambiae complex strains, with black pepper proving significantly more toxic than piperine. The An. funestus strains were substantially less sensitive to black pepper and piperine which may reflect a marked difference in the feeding habits of this species compared to that of the Gambiae complex or a difference in food metabolism as a consequence of differences in breeding habitat between species. Insecticide resistant and susceptible strains by species proved equally susceptible to black pepper and piperine. It is concluded that black pepper shows potential as a larvicide for the control of certain malaria vector species.

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

NARCIS (Netherlands)

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

2005-01-01

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

The effect of metal pollution on the life history and insecticide resistance phenotype of the major malaria vector Anopheles arabiensis (Diptera: Culicidae)

OpenAIRE

Oliver, Shüné V.; Brooke, Basil D.

2018-01-01

Metal exposure is one of the commonest anthropogenic pollutants mosquito larvae are exposed to, both in agricultural and urban settings. As members of the Anopheles gambiae complex, which contains several major malaria vector species including An. arabiensis, are increasingly adapting to polluted environments, this study examined the effects of larval metal exposure on various life history traits of epidemiological importance. Two laboratory strains of An. arabiensis, SENN (insecticide suscep...

Visualizing non infectious and infectious Anopheles gambiae blood feedings in naive and saliva-immunized mice.

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Valerie Choumet

Full Text Available 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

Visualizing non infectious and infectious Anopheles gambiae blood feedings in naive and saliva-immunized mice.

Science.gov (United States)

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

2012-01-01

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. 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. 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. 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.

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

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Gloria Volohonsky

2017-01-01

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

Habitat stability and occurrences of malaria vector larvae in western Kenya highlands

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Atieli Harrysone

2009-10-01

Full Text Available Abstract Background Although the occurrence of malaria vector larvae in the valleys of western Kenya highlands is well documented, knowledge of larval habitats in the uphill sites is lacking. Given that most inhabitants of the highlands actually dwell in the uphill regions, it is important to develop understanding of mosquito breeding habitat stability in these sites in order to determine their potential for larval control. Methods A total of 128 potential larval habitats were identified in hilltops and along the seasonal streams in the Sigalagala area of Kakamega district, western Kenya. Water availability in the habitats was followed up daily from August 3, 2006 to February 23, 2007. A habitat is defined as stable when it remains aquatic continuously for at least 12 d. Mosquito larvae were observed weekly. Frequencies of aquatic, stable and larvae positive habitats were compared between the hilltop and seasonal stream area using χ2-test. Factors affecting the presence/absence of Anopheles gambiae larvae in the highlands were determined using multiple logistic regression analysis. Results Topography significantly affected habitat availability and stability. The occurrence of aquatic habitats in the hilltop was more sporadic than in the stream area. The percentage of habitat occurrences that were classified as stable during the rainy season is 48.76% and 80.79% respectively for the hilltop and stream area. Corresponding frequencies of larvae positive habitats were 0% in the hilltop and 5.91% in the stream area. After the rainy season, only 23.42% of habitat occurrences were stable and 0.01% larvae positive habitats were found in the hilltops, whereas 89.75% of occurrences remained stable in the stream area resulting in a frequency of 12.21% larvae positive habitats. The logistic regression analysis confirmed the association between habitat stability and larval occurrence and indicated that habitat surface area was negatively affecting the

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

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Little Tom J

2009-06-01

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

Laboratory evaluation of three commercial coil products for protection efficacy against Anopheles gambiae from southern Ghana: a preliminary study.

Science.gov (United States)

Avicor, S W; Wajidi M, F F; Jaal, Z

2015-06-01

Residents in irrigated urban agricultural sites face numerous mosquito problems such as increased mosquito populations and reduced insecticides susceptibility due to the creation of mosquito breeding sites and agricultural use of insecticides and hence require effective protective products against them. In this study, the protection effectiveness of three pyrethroid formulated mosquito coils of Malaysian origin against Anopheles gambiae sensu lato from an irrigated urban agricultural site in Ghana were evaluated for their potential use. Sucrose fed An. gambiae s.l. were exposed to insecticide-containing coils in a 70 cm x 70 cm x 70 cm glass chamber to assess the insecticidal effect of the coils. The 0.005% metofluthrin coil caused the most rapid knockdown of 50% of the test mosquitoes. The mean lethal effect of the coils on An. gambiae s.l. were as follows; 0.005% metofluthrin (86%), 0.3% d-allethrin (74.33%), 0.15% d-trans allethrin (72%) and the 0.25% d-allethrin reference coil (69%). The 0.005% metofluthrin coil achieved the highest insecticidal effect on An. gambiae s.l. compared to the other coils and hence performed better than the others as an anti-mosquito product. All the three test coils were effective against An. gambaie s.l. from the irrigated agricultural site compared to the reference coil.

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

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João Pinto

2007-11-01

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

Insecticide susceptibility of natural populations of Anopheles coluzzii and Anopheles gambiae (sensu stricto) from Okyereko irrigation site, Ghana, West Africa.

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Chabi, Joseph; Baidoo, Philip K; Datsomor, Alex K; Okyere, Dora; Ablorde, Aikins; Iddrisu, Alidu; Wilson, Michael D; Dadzie, Samuel K; Jamet, Helen P; Diclaro, Joseph W

2016-03-31

The increasing spread of insecticide resistance in malaria vectors has been well documented across sub-Saharan Africa countries. The influence of irrigation on increasing vector resistance is poorly understood, and is critical to successful and ethical implementation of food security policies. This study investigated the insecticide resistance status of An. gambiae (s.l.) mosquitoes collected from the irrigated rice area of Okyereko, a village containing about 42 hectares of irrigated field within an irrigation project plan in the Central Region of Ghana. Large amounts of insecticides, herbicides and fertilizers are commonly used in the area to boost the annual production of the rice. Mosquito larvae were collected and adults were assayed from the F1 progeny. The resistance status, allele and genotype were characterized using WHO susceptibility testing and PCR methods respectively. The An. gambiae (s.l.) populations from Okyereko are highly resistant to DDT and pyrethroid insecticides, with possible involvement of metabolic mechanisms including the elevation of P450 and GST enzyme as well as P-gp activity. The population was mostly composed of An. coluzzii specimens (more than 96 %) with kdr and ace-1 frequencies of 0.9 and 0.2 %, respectively. This study brings additional information on insecticide resistance and the characterization of An. gambiae (s.l.) mosquitoes from Okyereko, which can be helpful in decision making for vector control programmes in the region.

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

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Janet M Meredith

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

Qualitative Study Of Anopheles Species In Konduga Lake Area Of ...

African Journals Online (AJOL)

The investigation on Anopheles species in Konduga lake area, Borno State of Nigeria, was carried out to identify various Anopheles species prevalent in the area and to determine their relative population densities. Six Anopheles species were recorded, namely, A. gambiae, A. funestus, A. ziemanni, A. squamosus, ...

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

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

2012-01-01

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

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

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

2012-04-17

In the last decade there have been marked reductions in malaria incidence in sub-Saharan Africa. Sustaining these reductions will rely upon insecticides to control the mosquito malaria vectors. We report that in the primary African malaria vector, Anopheles gambiae sensu stricto, a single enzyme, CYP6M2, confers resistance to two classes of insecticide. This is unique evidence in a disease vector of cross-resistance associated with a single metabolic gene that simultaneously reduces the efficacy of two of the four classes of insecticide routinely used for malaria control. The gene-expression profile of a highly DDT-resistant population of A. gambiae s.s. from Ghana was characterized using a unique whole-genome microarray. A number of genes were significantly overexpressed compared with two susceptible West African colonies, including genes from metabolic families previously linked to insecticide resistance. One of the most significantly overexpressed probe groups (false-discovery rate-adjusted P P450 gene CYP6M2. This gene is associated with pyrethroid resistance in wild A. gambiae s.s. populations) and can metabolize both type I and type II pyrethroids in recombinant protein assays. Using in vitro assays we show that recombinant CYP6M2 is also capable of metabolizing the organochlorine insecticide DDT in the presence of solubilizing factor sodium cholate.

Regulation of anti-Plasmodium immunity by a LITAF-like transcription factor in the malaria vector Anopheles gambiae.

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Ryan C Smith

Full Text Available The mosquito is the obligate vector for malaria transmission. To complete its development within the mosquito, the malaria parasite Plasmodium must overcome the protective action of the mosquito innate immune system. Here we report on the involvement of the Anopheles gambiae orthologue of a conserved component of the vertebrate immune system, LPS-induced TNFα transcription factor (LITAF, and its role in mosquito anti-Plasmodium immunity. An. gambiae LITAF-like 3 (LL3 expression is up-regulated in response to midgut invasion by both rodent and human malaria parasites. Silencing of LL3 expression greatly increases parasite survival, indicating that LL3 is part of an anti-Plasmodium defense mechanism. Electrophoretic mobility shift assays identified specific LL3 DNA-binding motifs within the promoter of SRPN6, a gene that also mediates mosquito defense against Plasmodium. Further experiments indicated that these motifs play a direct role in LL3 regulation of SRPN6 expression. We conclude that LL3 is a transcription factor capable of modulating SRPN6 expression as part of the mosquito anti-Plasmodium immune response.

Impact of agriculture on the selection of insecticide resistance in the malaria vector Anopheles gambiae: a multigenerational study in controlled conditions.

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Nkya, Theresia Estomih; Poupardin, Rodolphe; Laporte, Frederic; Akhouayri, Idir; Mosha, Franklin; Magesa, Stephen; Kisinza, William; David, Jean-Philippe

2014-10-16

Resistance of mosquitoes to insecticides is mainly attributed to their adaptation to vector control interventions. Although pesticides used in agriculture have been frequently mentioned as an additional force driving the selection of resistance, only a few studies were dedicated to validate this hypothesis and characterise the underlying mechanisms. While insecticide resistance is rising dramatically in Africa, deciphering how agriculture affects resistance is crucial for improving resistance management strategies. In this context, the multigenerational effect of agricultural pollutants on the selection of insecticide resistance was examined in Anopheles gambiae. An urban Tanzanian An. gambiae population displaying a low resistance level was used as a parental strain for a selection experiment across 20 generations. At each generation larvae were selected with a mixture containing pesticides and herbicides classically used in agriculture in Africa. The resistance levels of adults to deltamethrin, DDT and bendiocarb were compared between the selected and non-selected strains across the selection process together with the frequency of kdr mutations. A microarray approach was used for pinpointing transcription level variations selected by the agricultural pesticide mixture at the adult stage. A gradual increase of adult resistance to all insecticides was observed across the selection process. The frequency of the L1014S kdr mutation rose from 1.6% to 12.5% after 20 generations of selection. Microarray analysis identified 90 transcripts over-transcribed in the selected strain as compared to the parental and the non-selected strains. Genes encoding cuticle proteins, detoxification enzymes, proteins linked to neurotransmitter activity and transcription regulators were mainly affected. RT-qPCR transcription profiling of candidate genes across multiple generations supported their link with insecticide resistance. This study confirms the potency of agriculture in selecting

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

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Sylvie Cornelie

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

Quantification of the efficiency of treatment of Anopheles gambiae breeding sites with petroleum products by local communities in areas of insecticide resistance in the Republic of Benin

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Doannio Julien MC MC

2007-05-01

Full Text Available Abstract Background The emergence of Anopheles populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for developing new insecticidal molecules or, improving the efficacy of existing insecticides or abandoning those to which resistance has emerged. The use of petroleum products (PP against mosquito larvae had an immense success during early programmes of malaria control, but these compounds were abandoned and replaced in the 1950s by synthetic insecticides probably because of the high performances given by these new products. In the current context of vector resistance, it is important to elucidate the empirical use of PP by quantifying their efficiencies on resistant strains of Anopheles. Methods Larvae of Anopheles Ladji a local resistant strain were exposed to increasing concentrations of various PP (kerosene, petrol and engine oils for 24 hours and the lethal activities recorded. The highest concentration (HiC having no lethal activity (also referred as the NOEL or no effect level and the lowest concentration (LoC100 yielding 100% mortality were rated for each PP on the Ladji strain. Prior to laboratory analysis, KAP studies were conducted in three traditional communities were insecticide resistance is clearly established to confirm the use of PP against mosquitoes. Results Laboratory analysis of petrol, kerosene and engine oils, clearly established their lethal activities on resistant strains of Anopheles larvae. Contrary to existing references, this research revealed that exposed larvae of Anopheles were mostly killed by direct contact toxicity and not by suffocation as indicated in some earlier reports. Conclusion This research could serve as scientific basis to backup the empirical utilisation of PP on mosquito larvae and to envisage possibilities of using PP in some traditional settings where Anopheles have developed

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

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Ouédraogo Jean-Bosco

2008-09-01

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

Genetic Dissection of Anopheles gambiae Gut Epithelial Responses to Serratia marcescens

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Stathopoulos, Stavros; Neafsey, Daniel E.; Lawniczak, Mara K. N.; Muskavitch, Marc A. T.; Christophides, George K.

2014-01-01

Genetic variation in the mosquito Anopheles gambiae profoundly influences its ability to transmit malaria. Mosquito gut bacteria are shown to influence the outcome of infections with Plasmodium parasites and are also thought to exert a strong drive on genetic variation through natural selection; however, a link between antibacterial effects and genetic variation is yet to emerge. Here, we combined SNP genotyping and expression profiling with phenotypic analyses of candidate genes by RNAi-mediated silencing and 454 pyrosequencing to investigate this intricate biological system. We identified 138 An. gambiae genes to be genetically associated with the outcome of Serratia marcescens infection, including the peptidoglycan recognition receptor PGRPLC that triggers activation of the antibacterial IMD/REL2 pathway and the epidermal growth factor receptor EGFR. Silencing of three genes encoding type III fibronectin domain proteins (FN3Ds) increased the Serratia load and altered the gut microbiota composition in favor of Enterobacteriaceae. These data suggest that natural genetic variation in immune-related genes can shape the bacterial population structure of the mosquito gut with high specificity. Importantly, FN3D2 encodes a homolog of the hypervariable pattern recognition receptor Dscam, suggesting that pathogen-specific recognition may involve a broader family of immune factors. Additionally, we showed that silencing the gene encoding the gustatory receptor Gr9 that is also associated with the Serratia infection phenotype drastically increased Serratia levels. The Gr9 antibacterial activity appears to be related to mosquito feeding behavior and to mostly rely on changes of neuropeptide F expression, together suggesting a behavioral immune response following Serratia infection. Our findings reveal that the mosquito response to oral Serratia infection comprises both an epithelial and a behavioral immune component. PMID:24603764

The relationship between Anopheles gambiae density and rice cultivation in the savannah zone and forest zone of Côte d'Ivoire.

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Briët, Olivier J T; Dossou-Yovo, Joel; Akodo, Elena; van de Giesen, Nick; Teuscher, Thomas M

2003-05-01

In 13 villages in the savannah zone and 21 villages in the forest zone of Côte d'Ivoire, the biting density of the principal malaria vector, Anopheles gambiae, was studied as a function of rice cultivation in the inland valleys in a 2-km radius around each village. In the savannah villages, during the main season cropping period, surface water on rice-cultivated and to a lesser extent on uncultivated inland valleys seems to contribute strongly to the A. gambiae population density. For the off-season cropping period (which starts after the first light rains in the savannah zone), correlations were weaker. Breeding sites other than in inland valleys may play an important role in the savannah zone. In the forest zone, however, the A. gambiae population density was strongly correlated with the surface water availability (SWA) in the rice-cultivated inland valleys, whereas the correlation with the SWA in other (uncultivated) inland valleys was weak. The requirement of sunlit breeding sites for A. gambiae might explain this difference between zones. In the forest zone, only inland valleys cleared for rice cultivation meet this requirement, whereas all other inland valleys are covered with dense vegetation. In the savannah zone, however, most undergrowth is burnt during the dry season, which permits sunlight to reach puddles resulting from the first rains.

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

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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

Laboratory studies on the olfactory behaviour of Anopheles quadriannulatus

NARCIS (Netherlands)

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

2005-01-01

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

The susceptibility of five African Anopheles species to Anabaena PCC 7120 expressing Bacillus thuringiensis subsp. israelensis mosquitocidal cry genes

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Ketseoglou Irene

2012-10-01

Full Text Available Abstract Background Malaria, one of the leading causes of death in Africa, is transmitted by the bite of an infected female Anopheles mosquito. Problems associated with the development of resistance to chemical insecticides and concerns about the non-target effects and persistence of chemical insecticides have prompted the development of environmentally friendly mosquito control agents. The aim of this study was to evaluate the larvicidal activity of a genetically engineered cyanobacterium, Anabaena PCC 7120#11, against five African Anopheles species in laboratory bioassays. Findings There were significant differences in the susceptibility of the anopheline species to PCC 7120#11. The ranking of the larvicidal activity of PCC 7120#11 against species in the An. gambiae complex was: An. merus An. arabiensis An. gambiae An. quadriannulatus, where 50. The LC50 of PCC 7120#11 against the important malaria vectors An. gambiae and An. arabiensis was 12.3 × 105 cells/ml and 8.10 × 105 cells/ml, respectively. PCC 7120#11 was not effective against An. funestus, with less than 50% mortality obtained at concentrations as high as 3.20 × 107 cells/ml. Conclusions PCC 7120#11 exhibited good larvicidal activity against larvae of the An. gambiae complex, but relatively weak larvicidal activity against An. funestus. The study has highlighted the importance of evaluating a novel mosquitocidal agent against a range of malaria vectors so as to obtain a clear understanding of the agent’s spectrum of activity and potential as a vector control agent.

Repellent activity of the creams formulated from Annona senegalensis and Boswellia dalzielii leaf fractions and essential oils against Anopheles gambiae (Diptera: Culicidae

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Lame Younoussa

2016-12-01

Full Text Available Objective: To investigate the repellent efficacy of the creams formulated from methanol extract and n-hexane, chloroform, ethyl acetate and methanol fractions as well as essential oils of Annona senegalensis (A. senegalensis and Boswellia dalzielii (B. dalzielii leaves against the malarial vector Anopheles gambiae (An. gambiae in the laboratory. Methods: The efficacies of 25% w/w active ingredient creams formulated from the plant-based products were tested. Different concentrations of the creams, ranging from 2.0 to 12.0 mg/ cm²were applied on the exposed dorsal hand area (25 cm2 of volunteers. The treated hands were submitted to 50 caged blood-starved females of An. gambiae for 3 min after every 30 min until 180 min. Results: Total protection of up to 120 and 60 min without bites of An. gambiae were recorded with n-hexane creams applied at 12 mg/cm2 respectively for A. senegalensis and B. dalzielii. The essential oil creams of the two tested plants applied at 6 mg/cm2 protected volunteers up to 120 min without mosquito bites. The commercial Odomos cream (12% N,N-diethyl- 3-methylbenzamide tested as the positive control at 6 mg/cm2 protected volunteers from mosquito bites up to180 min. Conclusions: These results suggest that the cream formulated from the n-hexane fraction of A. senegalensis and essential oil creams of A. senegalensis and B. dalzielii leaves have the potential of a natural herbal source for the development of new, safe and eco-friendly repellent products to prevent An. gambiae bites.

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

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

2014-03-01

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

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

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

2005-01-01

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

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

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Walker Edward D

2005-03-01

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

EFEKTIVITAS Bacillus thuringiensis H-14 STRAIN LOKAL DALAM BUAH KELAPA TERHADAP LARVA Anopheles sp dan Culex sp di KAMPUNG LAUT KABUPATEN CILACAP

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Blondine Ch. P

2013-07-01

Full Text Available Abstrak Bacillus thuringiensis serotipe H-14 strain lokal adalah bakteri patogen bersifat target spesifiknya larva nyamuk, aman bagi mamalia dan lingkungan. Penelitian bertujuan menentukan efektivitas B. thuringiensis H-14 strain lokal yang dikembangbiakkan dalam buah kelapa untuk pengendalian larva Anopheles sp dan Culex sp. Rancangan eksperimental semu, terdiri dari kelompok perlakuan dan kontrol. Bacillus thuringiensis H-14 strain lokal dikembangbiakan dalam10 buah kelapa umur 6–8 bulan, dengan berat kira-kira 1 kg, telah berisi air kelapa sekitar 400-500 ml/buah kelapa yang diperoleh dari Desa Klaces, Kampung Laut, Kabupaten Cilacap. Diinkubasi selama 14 hari pada temperatur kamar dan ditebarkan di 6 kolam yang menjadi habitat perkembangbiakan larva nyamuk dengan luas berkisar 3–100 m2.Hasil yang diperoleh menunjukkan efektivitas B. thuringiensis H-14 strain lokal terhadap larva Anopheles sp dan Culex sp selama 1 hari sesudah penebaran kematian larva berturut-turut sebesar 80–100% dan 79,31–100%. Sedangkan pada hari ke-14 sebesar 69,30–76,71% dan 67,69–86,04%. Buah kelapa dapat digunakan sebagai media lokal alternatif untuk pengembangbiakan B. thuringiensis H-14 strain lokal Kata kunci: B. thuringiensis H-14,  strain  lokal, buah kelapa, pengendalian larva Abstract Bacillus thuringiensis serotype H-14 local strain is pathogenic bacteria which specific  target to mosquito larvae. It is safe for mammals and enviroment. The aims of this study was to determine the effectivity of B. thuringiensis H-14 local strain which culturing in thecoconut wates against Anopheles sp and Culex sp mosquito larvae. This research is quasi experiment which consist of treated  and control groups. Bacillus thuringiensis H-14 local strain was cultured in 10 coconuts with 6–8 months age with weight around 1 kg that contained were approximately 400-500 ml/coconut were taken from Klaces village, Kampung Laut. After that the coconuts incubated for 14

A High Affinity Adenosine Kinase from Anopheles gambiae

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Cassera, María B.; Ho, Meng-Chiao; Merino, Emilio F.; Burgos, Emmanuel S.; Rinaldo-Matthis, Agnes; Almo, Steven C.; Schramm, Vern L.

2011-01-01

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 (Km 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 Ap4A (2.0 Å resolution) reveals interactions for adenosine, ATP and the geometry for phosphoryl transfer. The polyphosphate charge is partly neutralized by a bound Mg2+ ion and an ion pair to a catalytic site Arg. The AgAK structure consists of a large catalytic core in a three-layered α/β/α sandwich, and a small cap domain in contact with adenosine. The specificity and tight-binding for adenosine arises 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 human adenosine kinase (48% identity) than to AK from Toxoplasma gondii (31% identity). With this extraordinary affinity for AgAK, adenosine is efficiently captured and converted to AMP at near the diffusion limit, suggesting an important role of this enzyme to maintain the adenine nucleotide pool. mRNA analysis verifies that AgAK transcripts are produced in the adult insects. PMID:21247194

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

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Fotsing Jean-Marie

2009-05-01

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

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

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Hans J. Overgaard

2014-12-01

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

1 SHORT COMMUNICATION Resting behaviour of Anopheles ...

African Journals Online (AJOL)

2011-10-04

Oct 4, 2011 ... Resting behaviour of Anopheles gambiae s.l. and its implication on malaria .... nature) were requested to sleep under the double mosquito net trap (one ... designed from the DNA sequences of the intergenic spacer region of ...

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

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Zhu, Lin; Qualls, Whitney A.; Marshall, John M; Arheart, Kris L.; DeAngelis, Donald L.; 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 houses.ConclusionsIncreases in densities of sugar sources or

Contributions of Anopheles larval control to malaria suppression in tropical Africa: review of achievements and potential.

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Walker, K; Lynch, M

2007-03-01

Malaria vector control targeting the larval stages of mosquitoes was applied successfully against many species of Anopheles (Diptera: Culicidae) in malarious countries until the mid-20th Century. Since the introduction of DDT in the 1940s and the associated development of indoor residual spraying (IRS), which usually has a more powerful impact than larval control on vectorial capacity, the focus of malaria prevention programmes has shifted to the control of adult vectors. In the Afrotropical Region, where malaria is transmitted mainly by Anopheles funestus Giles and members of the Anopheles gambiae Giles complex, gaps in information on larval ecology and the ability of An. gambiae sensu lato to exploit a wide variety of larval habitats have discouraged efforts to develop and implement larval control strategies. Opportunities to complement adulticiding with other components of integrated vector management, along with concerns about insecticide resistance, environmental impacts, rising costs of IRS and logistical constraints, have stimulated renewed interest in larval control of malaria vectors. Techniques include environmental management, involving the temporary or permanent removal of anopheline larval habitats, as well as larviciding with chemical or biological agents. This present review covers large-scale trials of anopheline larval control methods, focusing on field studies in Africa conducted within the past 15 years. Although such studies are limited in number and scope, their results suggest that targeting larvae, particularly in human-made habitats, can significantly reduce malaria transmission in appropriate settings. These approaches are especially suitable for urban areas, where larval habitats are limited, particularly when applied in conjunction with IRS and other adulticidal measures, such as the use of insecticide treated bednets.

PCR-based karyotyping of Anopheles gambiae inversion 2Rj identifies the BAMAKO chromosomal form

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Conway David J

2007-10-01

Full Text Available Abstract Background The malaria vector Anopheles gambiae is polymorphic for chromosomal inversions on the right arm of chromosome 2 that segregate nonrandomly between assortatively mating populations in West Africa. One such inversion, 2Rj, is associated with the BAMAKO chromosomal form endemic to southern Mali and northern Guinea Conakry near the Niger River. Although it exploits a unique ecology and both molecular and chromosomal data suggest reduced gene flow between BAMAKO and other A. gambiae populations, no molecular markers exist to identify this form. Methods To facilitate study of the BAMAKO form, a PCR assay for molecular karyotyping of 2Rj was developed based on sequences at the breakpoint junctions. The assay was extensively validated using more than 700 field specimens whose karyotypes were determined in parallel by cytogenetic and molecular methods. As inversion 2Rj also occurs in SAVANNA populations outside the geographic range of BAMAKO, samples were tested from Senegal, Cameroon and western Guinea Conakry as well as from Mali. Results In southern Mali, where 2Rj polymorphism in SAVANNA populations was very low and most of the 2Rj homozygotes were found in BAMAKO karyotypes, the molecular and cytogenetic methods were almost perfectly congruent. Elsewhere agreement between the methods was much poorer, as the molecular assay frequently misclassified 2Rj heterozygotes as 2R+j standard homozygotes. Conclusion Molecular karyotyping of 2Rj is robust and accurate on 2R+j standard and 2Rj inverted homozygotes. Therefore, the proposed approach overcomes the lack of a rapid tool for identifying the BAMAKO form across developmental stages and sexes, and opens new perspectives for the study of BAMAKO ecology and behaviour. On the other hand, the method should not be applied for molecular karyotyping of j-carriers within the SAVANNA chromosomal form.

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

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

2015-09-01

Aedes aegypti and Anopheles gambiae are two mosquito species that represent significant threats to global public health as vectors of Dengue virus and malaria parasites, respectively. Although mosquito populations have been effectively controlled through the use of synthetic insecticides, the emergence of widespread insecticide-resistance in wild mosquito populations is a strong motivation to explore new insecticidal chemistries. For these studies, Ae. aegypti and An. gambiae were treated with commercially available plant essential oils via topical application. The relative toxicity of each essential oil was determined, as measured by the 24-h LD(50) and percentage knockdown at 1 h, as compared with a variety of synthetic pyrethroids. For Ae. aegypti, the most toxic essential oil (patchouli oil) was ∼1,700-times less toxic than the least toxic synthetic pyrethroid, bifenthrin. For An. gambiae, the most toxic essential oil (patchouli oil) was ∼685-times less toxic than the least toxic synthetic pyrethroid. A wide variety of toxicities were observed among the essential oils screened. Also, plant essential oils were analyzed via gas chromatography/mass spectrometry (GC/MS) to identify the major components in each of the samples screened in this study. While the toxicities of these plant essential oils were demonstrated to be lower than those of the synthetic pyrethroids tested, the large amount of GC/MS data and bioactivity data for each essential oil presented in this study will serve as a valuable resource for future studies exploring the insecticidal quality of plant essential oils. © The Authors 2015. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.

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

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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 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 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 important when comparing two treatments that the same trapping device is used in both. The

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

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Amuzu, Hilaria; Wilson, Michael D; Boakye, Daniel A

2010-09-14

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. 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). 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 interruption of transmission in areas

Novel peptide marker corresponding to salivary protein gSG6 potentially identifies exposure to Anopheles bites.

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Anne Poinsignon

2008-06-01

Full Text Available In order to improve malaria control, and under the aegis of WHO recommendations, many efforts are being devoted to developing new tools for identifying geographic areas with high risk of parasite transmission. Evaluation of the human antibody response to arthropod salivary proteins could be an epidemiological indicator of exposure to vector bites, and therefore to risk of pathogen transmission. In the case of malaria, which is transmitted only by anopheline mosquitoes, maximal specificity could be achieved through identification of immunogenic proteins specific to the Anopheles genus. The objective of the present study was to determine whether the IgG response to the Anopheles gambiae gSG6 protein, from its recombinant form to derived synthetic peptides, could be an immunological marker of exposure specific to Anopheles gambiae bites.Specific IgG antibodies to recombinant gSG6 protein were observed in children living in a Senegalese area exposed to malaria. With the objective of optimizing Anopheles specificity and reproducibility, we designed five gSG6-based peptide sequences using a bioinformatic approach, taking into consideration i their potential antigenic properties and ii the absence of cross-reactivity with protein sequences of other arthropods/organisms. The specific anti-peptide IgG antibody response was evaluated in exposed children. The five gSG6 peptides showed differing antigenic properties, with gSG6-P1 and gSG6-P2 exhibiting the highest antigenicity. However, a significant increase in the specific IgG response during the rainy season and a positive association between the IgG level and the level of exposure to Anopheles gambiae bites was significant only for gSG6-P1.This step-by-step approach suggests that gSG6-P1 could be an optimal candidate marker for evaluating exposure to Anopheles gambiae bites. This marker could be employed as a geographic indicator, like remote sensing techniques, for mapping the risk of malaria. It could

Study on the species composition and ecology of anophelines in Addis Zemen, South Gondar, Ethiopia.

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Kindu, Mizan; Aklilu, Esayas; Balkew, Meshesha; Gebre-Michael, Teshome

2018-03-27

Malaria is a public health problem in Ethiopia and its transmission is generally unstable and seasonal. For the selection of the most appropriate vector control measures, knowledge on the ecology of the vector is necessary at a local level. Therefore, the objectives of this study were to document the species composition, breeding habitat characteristics and occurrence of anopheline larva in Sheni stream and the vectorial role of the prevailing Anopheles in relation to malaria transmission in Addis Zemen, Ethiopia. Immature anophelines were sampled from breeding habitats and characteristics, such as water temperature, turbidity, water current, water pH and other variables, of the habitats were measured from October 2011 to February 2012. Adult anophelines were sampled inside human dwellings using space spray and Center for Disease Control light traps. Artificial pit shelters and clay pots were also used for outdoor adult collections. Anophelines collected were identified using morphological key. The enzyme-linked immunosorbent assay was applied to detect circumsporozoite proteins of Plasmodium and source of blood meals. A total of 6258 Anopheles larvae were collected and identified morphologically. Five anopheline species were found: An. gambiae (s.l.), An. cinereus, An. demeilloni, An. christi and An. pretoriensis. Anopheles gambiae (s.l.) existed in most of the habitats investigated. Only the former three species were captured in the adult collections. Sun-lit Sheni stream, rain pools, hoof prints, drainage and irrigation canals were found to be habitats of larvae. Anopheles gambiae (s.l.) larvae were most abundantly sampled from sand mining and natural sand pools of Sheni stream. Multiple regression analysis showed that clear, permanent and temporary habitats devoid of mats of algae were the best predictors of An. gambiae (s.l.) larval abundance. It is also the responsible malaria vector in the study area and exhibits anthropophilic and endophagic behaviour. The

BIONOMY OF Anopheles punctulatus GROUP (Anopheles farauti, Anopheles koliensis, Anopheles punctulatus MALARIA VECTOR IN PAPUA PROVINCE

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Semuel Sandy

2014-08-01

Full Text Available ABSTRAKMalaria merupakan masalah kesehatan utama di Provinsi Papua dengan angka Annual Parasite Incidence (API padatahun 2011 sebesar 58 per 1000 penduduk dan Annual Malaria Incidence (AMI sebesar 169 per 1000 penduduk. Vektormalaria Papua dilaporkan Anopheles farauti, An. punctulatus dan An. koliensis. Tiga spesies tersebut aktif menggigit padamalam hari (nokturnal, antropofilik dengan karakteristik tempat perkembangbiakan, aktifitas menggigit, dan tempatistirahat dilaporkan spesifik setiap spesies. Kajian ini untuk melihat beberapa aspek bionomi (tempat perkembangbiakan,aktifitas menggigit, dan tempat istirahat. Larva An. farauti memiliki habitat di daerah pantai, perairan payau (memilikitoleransi terhadap salinitas 4,6%, irigasi buatan atau alami. Nyamuk dewasa An. farauti betina bersifat nokturnal,eksofagik, eksofilik, dan antropofilik. Larva An. koliensis tidak ditemukan di perairan payau, banyak ditemukan di hutanrawa, hutan sagu, kolam semi permanen atau permanen yang dangkal dan terpapar sinar matahari langsung. Nyamukdewasa An. koliensis bersifat nokturnal, antropofilik (78% menggigit manusia, eksofagik, eksofilik sedangkan larva An.punctulatus tidak ditemukan di air payau, tetapi ditemukan pada kolam dengan air jernih atau keruh dengan vegetasi atautanpa vegetasi air. Larva An. punctulatus juga ditemukan di hutan sagu dan hutan rawa dengan paparan sinar mataharilangsung. Nyamuk dewasa An. punctulatus bersifat nokturnal, antropofilik (98% menggigit manusia, eksofagik, endofilik.Data dasar mengenai perilaku nyamuk Anopheles (bionomi sangat diperlukan dalam mengembangkan pola intervensi dankontrol vektor yang lebih efektif dan efisien.Kata kunci: vektor malaria, An. punctulatus group, bionomi, PapuaABSTRACTMalaria is a major health problem in Papua province with Annual Parasite Incidence (API was reported 58/1000population in 2011, and the Annual Malaria Incidence (AMI was 169/1000 population. The malaria vector in Papua wereAnopheles

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

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

2014-08-01

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 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 meal, but decreased their egg output. These data provide evidence for a new role of Kir channels in mosquito fecundity, and further validates them as promising molecular targets for the development of a new class of mosquitocides to be used in vector control. Copyright © 2014 Elsevier Ltd. All rights reserved.

Comparing larvicidal Effect of Methanol Extract of the Aerial Parts of Henbane (Hyoscyamus niger L. and Oleander ( Nerium oleander L. plants on Anopheles spp Larvae (Diptera: Culicidae in Vitro

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Mahmoodreza Behravan

2017-03-01

Full Text Available Background Malaria is an infectious disease by fever and chills, anemia and splenomegaly genus Plasmodium parasite is the agent it. One of the easiest and least expensive methods to prevent this disease is removing the vector that usually by been done insecticides and chemical pesticides, but nowadays due to the damaging effects of by toxic chemicals is currently trying to organic toxic and plant compounds used to combat the pests. So in this study used from the Hyoscyamus niger L. and Nerium oleander L. to destroy the larvae of this vectors and positive results were compared these two plants together. Methods In this experimental study, H. niger and N. oleander collected and dried to extraction by methanol usingthe Rotary Evaporator. Mosquito larvae collected from stagnant water pits and ponds around the Birjand, Iran and order to apply the relevant identity tests and isolation of Anopheles spp mosquito larvae. Survival measurement were used to estimate LC50 values using Probit analysis in Excel 2010 and SPSS (ver 20 software. Results Both Hyoscyamus niger and Nerium oleander had positive effects on destroying the Anopheles spp larvae and between obtained results, the most effective extract for destroying the mosquitoes Anopheles spp larvae, was the flower extract of henbane (LC50 = 0/26 ppm and the weakest one, was the leaves extract of oleander (LC50 = 4/85 ppm. Conclusions According to the results, the flower extract of henbane is recommended as a toxic, organic and natural compounds to fight Anopheles spp mosquito larvae, which it is stronger and more effective compared to the other parts of these two plants.

Biological control of Anopheles darlingi, Aedes aegypti and Culex quinquefasciatus larvae using shrimps

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Willian Marinho Dourado Coelho

2017-08-01

Full Text Available Mosquitoes can act as vectors of important diseases such as malaria, dengue, Zika virus, yellow fever, Chikungunya and Mayaro fever, in addition to filariasis. The use of insecticides, larvicides, bed nets and repellents, besides the use of drugs as chemoprevention and the treatment of the sick are currently the pillars of the control of these vectors. We studied the biological control against of Anopheles darlingi, Aedes aegypti and Culex quinquefasciatus larvae using shrimps of the species M. pantanalense, M. amazonicum, M. brasiliense and M. jelskii. Larvae of mosquitoes were collected from the breeding environment and placed in a 500 and 1000 l tank containing 60 shrimps/m2. The predatory activity was evaluated for 30 days and, in all groups it was observed that 100% of the larvae were consumed in few minutes. In the environment, these same species of crustaceans were released in water bodies with the presence of larvae of these insects. In just 72 h there was a marked reduction of the larvae in the release sites of shrimps. Similarly, there was a reduction in the number of adult mosquitoes caught near the breeding sites, allowing to infer that, in places where the crustaceans were released, the predatory activity on the larvae of mosquitoes was sufficient to reduce the number of adult mosquitoes p ≤ 0,05. This is the first description of the predatory activity of M. pantanalense, M. amazonicum, M. brasiliense and M. jelskii on An. darlingi, A. aegypti and C. quinquefasciatus larvae, constituting an important tool of biological control of these parasites-vectors.

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

NARCIS (Netherlands)

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

2010-01-01

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

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

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Krzywinski Jaroslaw

2009-07-01

Full Text Available Abstract 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.

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

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Krzywinska, Elzbieta; Krzywinski, Jaroslaw

2009-07-06

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. 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. 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.

Molecular identification of Anopheles gambiae sensu stricto Giles ...

African Journals Online (AJOL)

User

2016-09-28

Sep 28, 2016 ... responsible for the transmission. The knowledge of which form, Anopheles coluzzii, A. .... National Council for Science and Technology and Health Research ..... Styer ML, Carey RJ, Wang JL, Scott TW (2007). Mosquitoes Do.

Screening for adulticidal bioactivity of South African plants against Anopheles arabiensis

CSIR Research Space (South Africa)

Maharaj, R

2011-08-01

Full Text Available of Corymbia citriodora (Myrtaceae), and leaves and seeds of both Ocimum kilimandscharicum and Ocimum suave (Lamiaceae) had exhibited a signifi- cant repellent effect against Anopheles gambiae s.s and An. arabiensis during application of plant material...

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

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Corine Ngufor

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

EKOLOGI Anopheles spp. DI KABUPATEN LOMBOK TENGAH

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Majematang Mading

2014-07-01

Full Text Available Malaria remains a public health problem in West Nusa Tenggara Province. Central Lombok District is one of the areas with high case of malaria. Annual Malaria Incidence (AMI was increased from 5.9 ‰ in 2006, 6.7‰ up to 8.1‰ in 2008. The objective of the study is to describe the ecological condition of Anopheles spp. through observation, measurement of environmental variables, larvae and adult collection. This research was an observational research with cross-sectional study. The population of this study is all mosquitos and breeding habitats of Anopheles spp. that exist in the research location. Ecological observations carried out on anopheles breeding habitats including acidity, salinity, shaded places and aquatic biota. Air temperature and humidity measured at the adult mosquitoes trapping sites. The result showed that pH values of water is around 9.00, salinity in the breeding habitats around 14 ppm, and water biota (i.e. moss, grass, shrimps, fishes, tadpoles and crabs surrounded by bushes with larvae density 0,1-28,8 each dipping. The air measurement at the time was between 23°-27° Celsius and 65%-84% humidity. This research concludes that ecology and environmental conditions were supporting the development of larvae and adult mosquito of Anopheles spp.Keywords: ecology, Anopheles spp., Central Lombok

Scanning electron microscopic (Sem studies on fourth instar larva and pupa of Anopheles (Cellia stephensi Liston (Anophelinae: Culicidae

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Jagbir Singh Kirti

2014-12-01

Full Text Available Anopheles (Cellia stephensi Liston is a major vector species of malaria in Indian subcontinent. Taxonomists have worked on its various morphological aspects and immature stages to explore additional and new taxonomic attributes. Scanning electron microscopic (SEM studies have been conducted on the fourth instar larva and pupa of An. stephensi to find additional taxonomic features for the first time from Punjab state.

Sama et al., Afr J Tradit Complement Altern Med. (2014) 11(4):84-88 ...

African Journals Online (AJOL)

cadewumi

BAILL AND BAILL, ON THE MALARIA VECTOR ANOPHELES GAMBIAE ... One of the programs designed to reduce the transmission of important vector-borne diseases, such as malaria, .... Larvae were fed with dog biscuit until they were.

Reflections on the Anopheles gambiae genome sequence, transgenic mosquitoes and the prospect for controlling malaria and other vector borne diseases.

Science.gov (United States)

Tabachnick, Walter J

2003-09-01

The completion of the Anopheles gambiae Giles genome sequencing project is a milestone toward developing more effective strategies in reducing the impact of malaria and other vector borne diseases. The successes in developing transgenic approaches using mosquitoes have provided another essential new tool for further progress in basic vector genetics and the goal of disease control. The use of transgenic approaches to develop refractory mosquitoes is also possible. The ability to use genome sequence to identify genes, and transgenic approaches to construct refractory mosquitoes, has provided the opportunity that with the future development of an appropriate genetic drive system, refractory transgenes can be released into vector populations leading to nontransmitting mosquitoes. An. gambiae populations incapable of transmitting malaria. This compelling strategy will be very difficult to achieve and will require a broad substantial research program for success. The fundamental information that is required on genome structure, gene function and environmental effects on genetic expression are largely unknown. The ability to predict gene effects on phenotype is rudimentary, particularly in natural populations. As a result, the release of a refractory transgene into natural mosquito populations is imprecise and there is little ability to predict unintended consequences. The new genetic tools at hand provide opportunities to address an array of important issues, many of which can have immediate impact on the effectiveness of a host of strategies to control vector borne disease. Transgenic release approaches represent only one strategy that should be pursued. A balanced research program is required.

Anthropogenic habitat disturbance and ecological divergence between incipient species of the malaria mosquito Anopheles gambiae.

Science.gov (United States)

Kamdem, Colince; Tene Fossog, Billy; Simard, Frédéric; Etouna, Joachim; Ndo, Cyrille; Kengne, Pierre; Boussès, Philippe; Etoa, François-Xavier; Awono-Ambene, Parfait; Fontenille, Didier; Antonio-Nkondjio, Christophe; Besansky, Nora J; Costantini, Carlo

2012-01-01

Anthropogenic habitat disturbance is a prime cause in the current trend of the Earth's reduction in biodiversity. Here we show that the human footprint on the Central African rainforest, which is resulting in deforestation and growth of densely populated urban agglomerates, is associated to ecological divergence and cryptic speciation leading to adaptive radiation within the major malaria mosquito Anopheles gambiae. In southern Cameroon, the frequency of two molecular forms--M and S--among which reproductive isolation is strong but still incomplete, was correlated to an index of urbanisation extracted from remotely sensed data, expressed as the proportion of built-up surface in each sampling unit. The two forms markedly segregated along an urbanisation gradient forming a bimodal cline of ∼6-km width: the S form was exclusive to the rural habitat, whereas only the M form was present in the core of densely urbanised settings, co-occurring at times in the same polluted larval habitats of the southern house mosquito Culex quinquefasciatus--a species association that was not historically recorded before. Our results indicate that when humans create novel habitats and ecological heterogeneities, they can provide evolutionary opportunities for rapid adaptive niche shifts associated with lineage divergence, whose consequences upon malaria transmission might be significant.

Insecticide resistance profile of Anopheles gambiae from a phase II field station in Cové, southern Benin: implications for the evaluation of novel vector control products.

Science.gov (United States)

Ngufor, Corine; N'Guessan, Raphael; Fagbohoun, Josias; Subramaniam, Krishanthi; Odjo, Abibatou; Fongnikin, Augustin; Akogbeto, Martin; Weetman, David; Rowland, Mark

2015-11-18

Novel indoor residual spraying (IRS) and long-lasting insecticidal net (LLIN) products aimed at improving the control of pyrethroid-resistant malaria vectors have to be evaluated in Phase II semi-field experimental studies against highly pyrethroid-resistant mosquitoes. To better understand their performance it is necessary to fully characterize the species composition, resistance status and resistance mechanisms of the vector populations in the experimental hut sites. Bioassays were performed to assess phenotypic insecticide resistance in the malaria vector population at a newly constructed experimental hut site in Cové, a rice growing area in southern Benin, being used for WHOPES Phase II evaluation of newly developed LLIN and IRS products. The efficacy of standard WHOPES-approved pyrethroid LLIN and IRS products was also assessed in the experimental huts. Diagnostic genotyping techniques and microarray studies were performed to investigate the genetic basis of pyrethroid resistance in the Cové Anopheles gambiae population. The vector population at the Cové experimental hut site consisted of a mixture of Anopheles coluzzii and An. gambiae s.s. with the latter occurring at lower frequencies (23 %) and only in samples collected in the dry season. There was a high prevalence of resistance to pyrethroids and DDT (>90 % bioassay survival) with pyrethroid resistance intensity reaching 200-fold compared to the laboratory susceptible An. gambiae Kisumu strain. Standard WHOPES-approved pyrethroid IRS and LLIN products were ineffective in the experimental huts against this vector population (8-29 % mortality). The L1014F allele frequency was 89 %. CYP6P3, a cytochrome P450 validated as an efficient metabolizer of pyrethroids, was over-expressed. Characterizing pyrethroid resistance at Phase II field sites is crucial to the accurate interpretation of the performance of novel vector control products. The strong levels of pyrethroid resistance at the Cové experimental hut

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

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Kerah-Hinzoumbé Clément

2009-09-01

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

Urban agriculture and Anopheles habitats in Dar es Salaam, Tanzania.

Science.gov (United States)

Dongus, Stefan; Nyika, Dickson; Kannady, Khadija; Mtasiwa, Deo; Mshinda, Hassan; Gosoniu, Laura; Drescher, Axel W; Fillinger, Ulrike; Tanner, Marcel; Killeen, Gerry F; Castro, Marcia C

2009-05-01

A cross-sectional survey of agricultural areas, combined with routinely monitored mosquito larval information, was conducted in urban Dar es Salaam, Tanzania, to investigate how agricultural and geographical features may influence the presence of Anopheles larvae. Data were integrated into a geographical information systems framework, and predictors of the presence of Anopheles larvae in farming areas were assessed using multivariate logistic regression with independent random effects. It was found that more than 5% of the study area (total size 16.8 km2) was used for farming in backyard gardens and larger open spaces. The proportion of habitats containing Anopheles larvae was 1.7 times higher in agricultural areas compared to other areas (95% confidence interval = 1.56-1.92). Significant geographic predictors of the presence of Anopheles larvae in gardens included location in lowland areas, proximity to river, and relatively impermeable soils. Agriculture-related predictors comprised specific seedbed types, mid-sized gardens, irrigation by wells, as well as cultivation of sugar cane or leafy vegetables. Negative predictors included small garden size, irrigation by tap water, rainfed production and cultivation of leguminous crops or fruit trees. Although there was an increased chance of finding Anopheles larvae in agricultural sites, it was found that breeding sites originated by urban agriculture account for less than a fifth of all breeding sites of malaria vectors in Dar es Salaam. It is suggested that strategies comprising an integrated malaria control effort in malaria-endemic African cities include participatory involvement of farmers by planting shade trees near larval habitats.

HUBUNGAN ANOPHELES BARBIROSTRIS DENGAN MALARIA

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Krisna Iryani

2013-03-01

Full Text Available Malaria is a disease caused by intercellular obligate protozoa genus of Plasmodium which is a parasite carried by female Anopheles mosquito. One of them is Anopheles barbirostris. Research in several places already proved that Anopheles barbirostris acts as a vector of malaria. One case that occurred in Cineam district, Tasikmalaya regency showed that Anopheles barbirostris is suspected as vector of malaria. This is proven through a research on the relationship between Anopheles barbirostris with malaria. Data was taken from the larvae and adult mosquitoes captured around Cineam village, Tasikmalaya. The observation was done in the open field and laboratory. Data and identification by pictorial key for female Anopheles showed that the population of Anopheles barbirostris was always a dominant population compared to another Anopheles species. Because of the breeding ponds and the resting places were around the village, it is suspected that they mainly bit humans. The result of the observation in laboratory showed the life cycle of Anopheles barbirostris are around 20-27 days, and the longevity of 20 days. Morphological identification of Anopheles barbirostris by pictorial key for female Anopheles showed that there is no any significant difference. This research showed that Anopheles barbirostris was suspected as vector of malaria in Cineam village, Tasikmalaya.

Insecticide susceptibility of Anopheles mosquitoes changes in response to variations in the larval environment.

Science.gov (United States)

Owusu, Henry F; Chitnis, Nakul; Müller, Pie

2017-06-16

Insecticide resistance threatens the success achieved through vector control in reducing the burden of malaria. An understanding of insecticide resistance mechanisms would help to develop novel tools and strategies to restore the efficacy of insecticides. Although we have substantially improved our understanding of the genetic basis of insecticide resistance over the last decade, we still know little of how environmental variations influence the mosquito phenotype. Here, we measured how variations in larval rearing conditions change the insecticide susceptibility phenotype of adult Anopheles mosquitoes. Anopheles gambiae and A. stephensi larvae were bred under different combinations of temperature, population density and nutrition, and the emerging adults were exposed to permethrin. Mosquitoes bred under different conditions showed considerable changes in mortality rates and body weight, with nutrition being the major factor. Weight is a strong predictor of insecticide susceptibility and bigger mosquitoes are more likely to survive insecticide treatment. The changes can be substantial, such that the same mosquito colony may be considered fully susceptible or highly resistant when judged by World Health Organization discriminatory concentrations. The results shown here emphasise the importance of the environmental background in developing insecticide resistance phenotypes, and caution for the interpretation of data generated by insecticide susceptibility assays.

Presence of Aedes and Anopheles mosquito larvae is correlated to bacteria found in domestic water-storage containers.

Science.gov (United States)

Nilsson, Louise K J; Sharma, Anil; Bhatnagar, Raj K; Bertilsson, Stefan; Terenius, Olle

2018-06-01

Water-storage containers are common in households where access to water is scarce and often act as breeding sites for vector mosquitoes. Bacteria in these containers may be important for attracting or repelling ovipositing mosquitoes. We hypothesized that bacterial community composition in water-storage containers would represent either inhibitory or suitable environmental conditions for mosquito larvae. To investigate this, we characterized the bacterial community composition in water-storage containers and correlated these communities to Aedes and Anopheles larval densities. Water samples were collected over two years from 13 containers in an Indian village and analyzed by high throughput 16S rRNA gene amplicon sequencing. Comparisons of bacterial community composition between water with and without mosquito larvae showed that Xanthomonadaceae, Comamonadaceae and Burkholderiaceae were more common (P < 0.05) in absence of larvae, while Lachnospiraceae, Synechococcaceae, Alcaligenaceae and Cryomorphaceae were more common (P < 0.05) in presence of larvae. Indicator analysis identified operational taxonomic units designated as CL500-29 marine group (Acidimicrobiaceae) and FukuN101 (Microbacteriaceae) for absence and presence of larvae, respectively. These results contribute to the understanding of which bacteria, directly or indirectly, can be linked to absence or presence of mosquitoes around households and set the basis for potential measures to be taken against these vector mosquitoes.

Field evaluation of synthetic lure (3-methyl-1-butanol) when compared to non odor-baited control in capturing Anopheles mosquitoes in varying land-use sites in Madagascar.

Science.gov (United States)

Zohdy, Sarah; Derfus, Kristin; Andrianjafy, Mbolatiana Tovo; Wright, Patricia C; Gillespie, Thomas R

2015-03-07

Malaria is the 4(th) largest cause of mortality in Madagascar. To better understand malaria transmission dynamics, it is crucial to map the distribution of the malaria vectors, mosquitoes belonging to the genus Anopheles. To do so, it is important to have a strong Anopheles-specific lure to ensure the maximum number of captures. Previous studies have isolated volatiles from the human skin microbiota and found the compound 3-methyl-1-butanol to be the most attractive to the malaria mosquito, Anopheles gambiae, in a laboratory setting; and recommended 3-methyl-1-butanol as a compound to increase An. gambiae captures in the field. To date, this compound's ability to lure wild mosquitoes in differing land-use settings has not been tested. In this study, we evaluate the role of the synthetic compound, 3-methyl-1-butanol in combination with field produced CO(2) in attracting Anopheles mosquitoes in varying land-use sites in Madagascar. CDC miniature light traps in combination with field produced CO(2) were deployed in and around six villages near Ranomafana National Park, Madagascar. To test the role of 3-methyl-1-butanol in luring Anopheles mosquitoes, two traps were set in each land-use site (village, agricultural sites, and forested habitats affiliated with each village). One was baited with the synthetic odor and the other was kept as a non-baited control. While 3-methyl-1-butanol baited traps did capture An. gambiae s.l. in this study, we did not find traps baited with synthetic 3-methyl-1-butanol to be more successful in capturing Anopheles mosquitoes, (including Anopheles gambiae s.l.) than the non odor-baited control traps in any of the land-use sites examined; however, regardless of odor bait, trapping near livestock pens resulted in the capture of significantly more Anopheles specimens. A strong synthetic lure in combination with insecticide has great potential as a mosquito control. Our findings suggest that trapping mosquitoes near livestock in malaria

Decline in frequency of the 2La chromosomal inversion in Anopheles gambiae (s.s.) in Western Kenya: correlation with increase in ownership of insecticide-treated bed nets.

Science.gov (United States)

Matoke-Muhia, Damaris; Gimnig, John E; Kamau, Luna; Shililu, Josephat; Bayoh, M Nabie; Walker, Edward D

2016-06-10

The 2La chromosomal inversion, a genetic polymorphism in An. gambiae (sensu stricto) (s.s.), is associated with adaptation to microclimatic differences in humidity and desiccation resistance and mosquito behaviors. Ownership of insecticide-treated bed nets (ITNs) for malaria control has increased markedly in western Kenya in the last 20 years. An increase in the frequency of ITNs indoors could select against house entering or indoor resting of Anopheles mosquitoes. Thus, the frequency of the 2La inversion is postulated to change in An. gambiae (s.s.) with the increase of ITN ownership over time. Anopheles gambiae mosquitoes were sampled between 1994 and 2011 using pyrethrum knockdown, bednet traps and human landing catches (HLC) from Asembo and Seme, western Kenya. The 2La inversion was detected by a PCR assay with primers designed for proximal breakpoints of the 2La/a and 2L+(a)/+(a) chromosomal conformation. Mosquitoes were tested for malaria parasite infection by sporozoite ELISA. The frequency of the 2La chromosomal inversion declined from 100 % of all chromosomes in 1994 to 17 % in 2005 and remained low through 2011 (21 %). ITN ownership increased from 0 to > 90 % of houses in the study area during this interval. The decline in the frequency of the 2La chromosomal inversion was significantly, negatively correlated with year (r = -0.93) and with increase in ITN ownership (r = -0.96). The frequency of the homo- and heterokaryotypes departed significantly from Hardy-Weinberg equilibrium, suggesting that 2La/a karyotype was under selection, earlier in its favor and later, against it. Precipitation and maximum monthly temperature did not vary over time, therefore there was no trend in climate that could account for the decline. There was no significant difference in frequency of the 2La inversion in An. gambiae (s.s.) females sampled indoors or outdoors in HCL in 2011, nor was there an association between the 2La inversion and infection with Plasmodium

Evaluación de la actividad insecticida de un producto granulado a base de Bacillus sphaericus sobre larvas de Anopheles albimanus y Culex quinquefasciatus en condiciones experimentales

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Estrella Cárdenas

2009-01-01

Full Text Available Culex quinquefasciatus causa alergias por las picaduras y ha sido asociado con la encefalitis equina, mientras que Anopheles albimanus es vector de malaria. El objetivo fue evaluar la toxicidad de un producto granulado a base de Bacillus sphaericus sobre larvas de Culex quinquefasciatus y Anopheles albimanus. El estudio fue realizado sobre larvas de C. quinquefasciatus (Sibaté y Villavicencio y An albimanus (Cartagena y Barranquilla. Las concentraciones ensayadas de B. sphaericus fueron: 20, 40, 60, 80, 100, 120, 140, 160, 180, 200 y 500 ppm para larvas de An. albimanus y para larvas de C. quinquefasciatus fueron: 2, 4, 6, 8, 10, 12, 14 y 16 ppm. Se utilizaron 20 larvas en tres repeticiones por cada concentración del producto y el tiempo de exposición fue de 48 horas. El producto de B. sphaericus mostró alta mortalidad (entre 80 y 100% a bajas concentraciones (8 y 12 ppm para larvas de C. quinquefasciatus; mientras que para larvas de An. albimanus a concentraciones más altas (entre 40 y 200 ppm la mortalidad no superó el 50%. La CL50 Logit fue 176 ppm y la Probit fue 192 ppm de B. sphaericus para larvas de An. albimanus; mientras que para larvas de C. quinquefasciatus la CL50 Logit fue 1, 9 ppm y la Probit fue 2, 2 ppm. Se concluye que las larvas de C. quinquefasciatus fueron más susceptibles a bajas concentraciones (2 a 12 ppm de B. sphaericus, mientras que las larvas de An. albimanus mostraron alta toxicidad a concentraciones de 500 ppm.

Ecology of Anopheles spp. in Central Lombok Regency

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Majematang Mading

2014-06-01

Full Text Available Malaria remains a public health problem in West Nusa Tenggara Province. Central Lombok District is one of the areas with high case of malaria. Annual Malaria Incidence (AMI was increased from 5.9 ‰ in 2006, 6.7‰ up to 8.1‰ in 2008. The objective of the study is to describe the ecological condition of Anopheles spp. through observation, measurement of environmental variables, larvae and adult collection. This research was an observational research with cross-sectional study. The population of this study is all mosquitos and breeding habitats of Anopheles spp. that exist in the research location. Ecological observations carried out on anopheles breeding habitats including acidity, salinity, shaded places and aquatic biota. Air temperature and humidity measured at the adult mosquitoes trapping sites. The result showed that pH values of water is around 9.00, salinity in the breeding habitats around 14 ppm, and water biota (i.e. moss, grass, shrimps, fishes, tadpoles and crabs surrounded by bushes with larvae density 0,1-28,8 each dipping. The air measurement at the time was between 23°-27° Celsius and 65%-84% humidity. This research concludes that ecology and environmental conditions were supporting the development of larvae and adult mosquito of Anopheles spp.

Insecticide resistance testing in malaria vectors in Tanzania ...

African Journals Online (AJOL)

mosquito survived much better and the scientists had a total of 467 mosquitoes to run the insecticide susceptibility tests. Innovative ways are necessary under field conditions for mosquito breeding in susceptibility studies. Key words: Malaria, Anopheles gambiae complex, larvae, fabric, resistance, susceptibility, Tanzania.

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

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Jackson M Muema

Full Text Available Successful optimization of plant-derived compounds into control of nuisance insects would benefit from scientifically validated targets. However, the close association between the genotypic responses and physiological toxicity effects mediated by these compounds remains underexplored. In this study, we evaluated the sublethal dose effects of proanthocyanidins (PAs sourced from green tea (Camellia sinensis on life history traits of Anopheles gambiae (sensu stricto mosquitoes with an aim to unravel the probable molecular targets. Based on the induced phenotypic effects, genes selected for study targeted juvenile hormone (JH biosynthesis, signal transduction, oxidative stress response and xenobiotic detoxification in addition to vitellogenesis in females. Our findings suggest that chronic exposure of larval stages (L3/L4 to sublethal dose of 5 ppm dramatically extended larval developmental period for up to 12 days, slowed down pupation rates, induced abnormal larval-pupal intermediates and caused 100% inhibition of adult emergence. Further, females exhibited significant interference of fecundity and egg hatchability relative to controls (p < 0.001. Using reverse transcription quantitative polymerase chain reaction (RT-qPCR, our findings show that PA-treated larvae exhibited significant repression of AgamJHAMT (p < 0.001, AgamILP1 (p < 0.001 and AgamCYP6M2 (p < 0.001 with up-regulation of Hsp70 (p < 0.001. Females exposed as larvae demonstrated down-regulation of AgamVg (p = 0.03, AgamILP1 (p = 0.009, AgamCYP6M2 (p = 0.05 and AgamJHAMT (p = 0.02. Our findings support that C. sinensis proanthocyanidins affect important vectorial capacity components such as mosquito survival rates and reproductive fitness thus could be potentially used for controlling populations of malaria vectors.

Age and prior blood feeding of Anopheles gambiae influences their susceptibility and gene expression patterns to ivermectin-containing blood meals.

Science.gov (United States)

Seaman, Jonathan A; Alout, Haoues; Meyers, Jacob I; Stenglein, Mark D; Dabiré, Roch K; Lozano-Fuentes, Saul; Burton, Timothy A; Kuklinski, Wojtek S; Black, William C; Foy, Brian D

2015-10-15

Ivermectin has been proposed as a novel malaria transmission control tool based on its insecticidal properties and unique route of acquisition through human blood. To maximize ivermectin's effect and identify potential resistance/tolerance mechanisms, it is important to understand its effect on mosquito physiology and potential to shift mosquito population age-structure. We therefore investigated ivermectin susceptibility and gene expression changes in several age groups of female Anopheles gambiae mosquitoes. The effect of aging on ivermectin susceptibility was analyzed in three age groups (2, 6, and 14-days) of colonized female Anopheles gambiaemosquitoes using standard survivorship assays. Gene expression patterns were then analyzed by transcriptome sequencing on an Illumina HiSeq 2500 platform. RT-qPCR was used to validate transcriptional changes and also to examine expression in a different, colonized strain and in wild mosquitoes, both of which blood fed naturally on an ivermectin-treated person. Mosquitoes of different ages and blood meal history died at different frequencies after ingesting ivermectin. Mortality was lowest in 2-day old mosquitoes exposed on their first blood meal and highest in 6-day old mosquitoes exposed on their second blood meal. Twenty-four hours following ivermectin ingestion, 101 and 187 genes were differentially-expressed relative to control blood-fed, in 2 and 6-day groups, respectively. Transcription patterns of select genes were similar in membrane-fed, colonized, and naturally-fed wild vectors. Transcripts from several unexpected functional classes were highly up-regulated, including Niemann-Pick Type C (NPC) genes, peritrophic matrix-associated genes, and immune-response genes, and these exhibited different transcription patterns between age groups, which may explain the observed susceptibility differences. Niemann-Pick Type 2 genes were the most highly up-regulated transcripts after ivermectin ingestion (up to 160 fold) and

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

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Clare C McArthur

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

Larvicidal properties of Lepidagathis alopecuroides and Azadirachta ...

African Journals Online (AJOL)

GRACE

2006-05-02

May 2, 2006 ... Comparative analysis of the larvicidal properties of aqueous extracts of leaves of Lepidagathis alopecuroides and Azadirachta indica (neem) was carried out on Anopheles gambiae and Culex quinquefasciatus. Assays showed that L. alopecuroides was more toxic to both larvae, while C. quinquefasciatus ...

Field evaluation of spatial repellency of metofluthrin-impregnated plastic strips against Anopheles gambiae complex in Bagamoyo, coastal Tanzania.

Science.gov (United States)

Kawada, Hitoshi; Temu, Emmanuel A; Minjas, Japhet N; Matsumoto, Osamu; Iwasaki, Tomonori; Takagi, Masahiro

2008-09-01

Metofluthrin is a newly synthesized pyrethroid possessing high knockdown and lethal activity against mosquitoes. Studies of metofluthrin-impregnated plastic strips have been performed with dengue vectors. This study reports the efficacy of the new prototypes of metofluthrin-impregnated plastic strips against malaria vectors, Anopheles gambiae complex, in the Kongo villages of Bagamoyo district in coastal Tanzania. The study, using 20 houses, half intervention, half control, was conducted for a 124-day period. Pyrethrum spray sheets and Centers for Disease Control and Prevention light traps were used to sample mosquito population indices. The mosquito density indices of the intervention houses were observed to be significantly lower than those of the control houses when pyrethrum spray sheet collection was used (F = 4.61, 1 df, P = 0.038; 98.7% reduction of total mosquito collection compared with that for the controls). These low indices were observed despite the large openings found in Bagamoyo houses, which were predicted to have a considerable negative effect on the spatial repellency of metofluthrin. Based on the present results, the pyrethrum spray sheet collection was the better of the 2 collection methods.

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

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Ingham, Victoria A; Jones, Christopher M; Pignatelli, Patricia; Balabanidou, Vasileia; Vontas, John; Wagstaff, Simon C; Moore, Jonathan D; Ranson, Hilary

2014-11-25

The elevated expression of enzymes with insecticide metabolism activity can lead to high levels of insecticide resistance in the malaria vector, Anopheles gambiae. In this study, adult female mosquitoes from an insecticide susceptible and resistant strain were dissected into four different body parts. RNA from each of these samples was used in microarray analysis to determine the enrichment patterns of the key detoxification gene families within the mosquito and to identify additional candidate insecticide resistance genes that may have been overlooked in previous experiments on whole organisms. A general enrichment in the transcription of genes from the four major detoxification gene families (carboxylesterases, glutathione transferases, UDP glucornyltransferases and cytochrome P450s) was observed in the midgut and malpighian tubules. Yet the subset of P450 genes that have previously been implicated in insecticide resistance in An gambiae, show a surprisingly varied profile of tissue enrichment, confirmed by qPCR and, for three candidates, by immunostaining. A stringent selection process was used to define a list of 105 genes that are significantly (p ≤0.001) over expressed in body parts from the resistant versus susceptible strain. Over half of these, including all the cytochrome P450s on this list, were identified in previous whole organism comparisons between the strains, but several new candidates were detected, notably from comparisons of the transcriptomes from dissected abdomen integuments. The use of RNA extracted from the whole organism to identify candidate insecticide resistance genes has a risk of missing candidates if key genes responsible for the phenotype have restricted expression within the body and/or are over expression only in certain tissues. However, as transcription of genes implicated in metabolic resistance to insecticides is not enriched in any one single organ, comparison of the transcriptome of individual dissected body parts cannot

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

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Ranson Hilary

2008-11-01

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

Hydrological modeling of geophysical parameters of arboviral and protozoan disease vectors in Internally Displaced People camps in Gulu, Uganda.

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Jacob, Benjamin G; Muturi, Ephantus J; Caamano, Erick X; Gunter, James T; Mpanga, Enoch; Ayine, Robert; Okelloonen, Joseph; Nyeko, Jack Pen-Mogi; Shililu, Josephat I; Githure, John I; Regens, James L; Novak, Robert J; Kakoma, Ibulaimu

2008-03-14

The aim of this study was to determine if remotely sensed data and Digital Elevation Model (DEM) can test relationships between Culex quinquefasciatus and Anopheles gambiae s.l. larval habitats and environmental parameters within Internally Displaced People (IDP) campgrounds in Gulu, Uganda. A total of 65 georeferenced aquatic habitats in various IDP camps were studied to compare the larval abundance of Cx. quinquefasciatus and An. gambiae s.l. The aquatic habitat dataset were overlaid onto Land Use Land Cover (LULC) maps retrieved from Landsat imagery with 150 m x 150 m grid cells stratified by levels of drainage. The LULC change was estimated over a period of 14 years. Poisson regression analyses and Moran's I statistics were used to model relationships between larval abundance and environmental predictors. Individual larval habitat data were further evaluated in terms of their covariations with spatial autocorrelation by regressing them on candidate spatial filter eigenvectors. Multispectral QuickBird imagery classification and DEM-based GIS methods were generated to evaluate stream flow direction and accumulation for identification of immature Cx. quinquefasciatus and An. gambiae s.l. and abundance. The main LULC change in urban Gulu IDP camps was non-urban to urban, which included about 71.5 % of the land cover. The regression models indicate that counts of An. gambiae s.l. larvae were associated with shade while Cx. quinquefasciatus were associated with floating vegetation. Moran's I and the General G statistics for mosquito density by species and instars, identified significant clusters of high densities of Anopheles; larvae, however, Culex are not consistently clustered. A stepwise negative binomial regression decomposed the immature An. gambiae s.l. data into empirical orthogonal bases. The data suggest the presence of roughly 11% to 28 % redundant information in the larval count samples. The DEM suggest a positive correlation for Culex (0.24) while for

EFICACIA DEL BACILLUS SPHAERICUS 2362 EN EL CONTROL DE LARVAS DE ANOPHELES PSUDOPUNCTIPENNIS (THEOBALD, 1901 Y CULEX QUINQUEFASCIATUS (SAY, 1823 EN BIOENSAYO DE LABORATORIO

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Diana Nongrados

2014-06-01

Full Text Available Se probó la actividad del Bacillus sphaericus 2362 en formulación líquida contra larvas de Anopheles pseudopunctipennis (Theobald, 1901 y Culex quinquefasciatus (Say, 1823 en bioensayos de laboratorio. Se utilizó agua destilada y agua de criadero, con concentraciones de 1 x 10 11, 1,5 x 10 11, 2,5 x 10 11, 3,9 x 10 11 esporas/mL. La población blanco se mantuvo en condiciones de laboratorio, usándose agua y flora nativa de criadero. Se vertieron dosis de 1 mL en vasos de prueba que contenían 25 larvas del II y III estadio y 150 mL de agua de criadero o agua destilada respectivamente; se usaron vasos con igual cantidad de agua y larvas como controles para cada concentración, tipo de agua y especie de larva probada. Las pruebas fueron realizadas tres veces para cada concentración, en condiciones de laboratorio. Se realizaron 12 réplicas divididas en 6 para cada especie, usándose un total de 4,800 larvas por especie. Las lecturas de la mortalidad de larvas fueron a las 12, 24, 48 y 72 h después de añadido Bacillus sphaericus. Se observó la elevada susceptibilidad de Culex quinquefasciatus a Bacillus sphaericus 2362, con una mortalidad mayor al 90% cuando se compararon los grupos tratados y controles (valor de p = 0,031 y 0,012 para cada tipo de agua respectivamente a las 48 h y con una concentración de 1,5 x 10 11 esporas/mL. Se demostró ampliamente que Anopheles pseudopunctipennis no es susceptible a Bacillus sphaericus 2362 en bioensayos de laboratorio y no se encontraron diferencias significativas en los tratamientos con diferentes tipos de agua (valor de p > 0,05.

Control of pyrethroid and DDT-resistant Anopheles gambiae by application of indoor residual spraying or mosquito nets treated with a long-lasting organophosphate insecticide, chlorpyrifos-methyl

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Chabi Joseph

2010-02-01

Full Text Available Abstract Background Scaling up of long-lasting insecticidal nets (LLINs and indoor residual spraying (IRS with support from the Global Fund and President's Malaria Initiative is providing increased opportunities for malaria control in Africa. The most cost-effective and longest-lasting residual insecticide DDT is also the most environmentally persistent. Alternative residual insecticides exist, but are too short-lived or too expensive to sustain. Dow Agrosciences have developed a microencapsulated formulation (CS of the organophosphate chlorpyrifos methyl as a cost-effective, long-lasting alternative to DDT. Methods Chlorpyrifos methyl CS was tested as an IRS or ITN treatment in experimental huts in an area of Benin where Anopheles gambiae and Culex quinquefasiactus are resistant to pyrethroids, but susceptible to organophosphates. Efficacy and residual activity was compared to that of DDT and the pyrethroid lambdacyalothrin. Results IRS with chlorpyrifos methyl killed 95% of An. gambiae that entered the hut as compared to 31% with lambdacyhalothrin and 50% with DDT. Control of Cx. quinquefasciatus showed a similar trend; although the level of mortality with chlorpyrifos methyl was lower (66% it was still much higher than for DDT (14% or pyrethroid (15% treatments. Nets impregnated with lambdacyhalothrin were compromized by resistance, killing only 30% of An. gambiae and 8% of Cx. quinquefasciatus. Nets impregnated with chlorpyrifos methyl killed more (45% of An gambiae and 15% of Cx. quinquefasciatus, but its activity on netting was of short duration. Contact bioassays on the sprayed cement-sand walls over the nine months of monitoring showed no loss of activity of chlorpyrifos methyl, whereas lambdacyhalothrin and DDT lost activity within a few months of spraying. Conclusion As an IRS treatment against pyrethroid resistant mosquitoes chlorpyrifos methyl CS outperformed DDT and lambdacyhalothrin. In IRS campaigns, chlorpyrifos methyl CS should

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

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Sutcliffe Alice C

2009-10-01

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

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

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He Ningjia

2008-01-01

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

Larvicidal Activity of Centaurea bruguierana ssp. belangerana Against Anopheles stephensi Larvae.

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Khanavi, Mahnaz; Rajabi, Afsaneh; Behzad, Masoud; Hadjiakhoondi, Abbas; Vatandoost, Hassan; Abaee, Mohammad Reza

2011-01-01

In this study, the total 80% of MeOH extract and also petroleum ether, CHCl3, EtOAc, n-BuOH, and the remaining MeOH fractions obtained by solvent-solvent fractionation of the whole flowering samples of Centaurea bruguierana (DC.) Hand.-Mzt. ssp. belangerana (DC.) Bornm. (Asteraceae), namely "Baad-Avard", collected from Borazjan in Bushehr Province (Bushehr, Iran) were investigated for larvicidal activity against malaria vector, Anopheles stephensi Liston, according to WHO methods. The mortality rate of total extract and petroleum ether fraction in concentration of 40 ppm were 28% and 86% respectively and the other fractions were inactive. The probit regression analysis for the dose-response to petroleum ether fraction treatment of larvae exhibited the LC50 and LC90 values of 15.7 ppm and 48.3 ppm, respectively. As results showed, the larvicidal activity of the petroleum ether fraction would be due to the nonpolar compounds in the plant which further isolation and purification would obtain the more active compounds in lower concentrations useful for preparation of biological insecticides.

Vectorial status and insecticide resistance of Anopheles funestus from a sugar estate in southern Mozambique.

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Kloke, R Graham; Nhamahanga, Eduardo; Hunt, Richard H; Coetzee, Maureen

2011-02-09

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. 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. 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 estate for seven years and the

Vectorial status and insecticide resistance of Anopheles funestus from a sugar estate in southern Mozambique

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Nhamahanga Eduardo

2011-02-01

Full Text Available Abstract 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

Efficacy of PermaNet® 3.0 and PermaNet® 2.0 nets against laboratory-reared and wild Anopheles gambiae sensu lato populations in northern Tanzania.

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Kweka, Eliningaya J; Lyaruu, Lucile J; Mahande, Aneth M

2017-01-18

Mosquitoes have developed resistance against pyrethroids, the only class of insecticides approved for use on long-lasting insecticidal nets (LLINs). The present study sought to evaluate the efficacy of the pyrethroid synergist PermaNet® 3.0 LLIN versus the pyrethroid-only PermaNet® 2.0 LLIN, in an East African hut design in Lower Moshi, northern Tanzania. In this setting, resistance to pyrethroid insecticides has been identified in Anopheles gambiae mosquitoes. Standard World Health Organization bioefficacy evaluations were conducted in both laboratory and experimental huts. Experimental hut evaluations were conducted in an area where there was presence of a population of highly pyrethroid-resistant An. arabiensis mosquitoes. All nets used were subjected to cone bioassays and then to experimental hut trials. Mosquito mortality, blood-feeding inhibition and personal protection rate were compared between untreated nets, unwashed LLINs and LLINs that were washed 20 times. Both washed and unwashed PermaNet® 2.0 and PermaNet® 3.0 LLINs had knockdown and mortality rates of 100% against a susceptible strain of An. gambiae sensu stricto. The adjusted mortality rate of the wild mosquito population after use of the unwashed PermaNet® 3.0 and PermaNet® 2.0 nets was found to be higher than after use of the washed PermaNet® 2.0 and PermaNet® 3.0 nets. Given the increasing incidence of pyrethroid resistance in An. gambiae mosquitoes in Tanzania, we recommend that consideration is given to its distribution in areas with pyrethroid-resistant malaria vectors within the framework of a national insecticide-resistance management plan.

Lack of insecticidal effect of mosquito coils containing either metofluthrin or esbiothrin on Anopheles gambiae sensu lato mosquitoes.

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Lukwa, Nzira; Chiwade, Tonderai

2008-12-01

Use of mosquito coils for personal protection against malaria and mosquito nuisance is advocated under mosquito and malaria control programmes. We performed field studies of mosquito coils containing either metofluthrin or esbiothrin in experimental huts situated in Kamhororo village, Gokwe district, Zimbabwe. All tests were performed on 3-5 day old reared female Anopheles gambiae sensu lato mosquitoes. The burning times were 9hr 20min for mosquito coils containing metofluthrin and 8 hr for those containing esbiothrin and the results were significantly different (p = metofluthrin was 90% and that for esbiothrin was 73.3% and the results were significantly different (p = 0.00). Mosquito coils containing metofluthrin had a mean repellence of 92.7% as compared to 85.4% for esbiothrin and the results were not significantly different (p=0.27). The protection time as required by EPA (1999) was 6 hr for mosquito coils containing metofluthrin and 5 hr for those containing esbiothrin. The mean insecticidal effect of mosquito coils containing metofluthrin was 84% as compared to 83% for those containing esbiothrin and the results were not significantly different (p = 0.56). Both mosquito formulations could not be classified as having insecticidal effect since none of them met the 95% mortality rate criteria.

Mosquitocidal Activity and Mode of Action of the Isoxazoline Fluralaner

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Jiang, Shiyao; Tsikolia, Maia; Bernier, Ulrich R.; Bloomquist, Jeffrey R.

2017-01-01

Mosquitoes, such as Aedes aegypti and Anopheles gambiae, are important vectors of human diseases. Fluralaner, a recently introduced parasiticide, was evaluated as a mosquitocide in this study. On Ae. aegypti and An. gambiae fourth-instar larvae, fluralaner had 24-h LC50 (lethal concentration for 50% mortality) values of 1.8 ppb and 0.4 ppb, respectively. Following topical application to adult Ae. aegypti, fluralaner toxicity reached a plateau in about 3 days, with 1- and 3-day LD50 (lethal do...

Larvicidal effects of leaf, bark and nutshell of Anacardium ...

African Journals Online (AJOL)

Comparative analysis of the larvicidal properties of aqueous extracts of leaves, bark and nutshell of Anacardium occidentale L. (Cashew) were evaluated on the larvae of Anopheles gambiae. Three concentrations of 10/100ml, 20/100ml and 30/100ml each of leaf, bark and nutshell were prepared in three replicates.

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

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Zhao Qin

2005-09-01

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

Development of a DNA-Based Method for Distinguishing the Malaria Vectors, Anopheles gambiae From Anopheles arabiensis.

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1986-06-01

our preliminary studies hybridization with the Droso- phila actin probe required such low stringency conditions that the signal to noise ratio made...Balabacensis complex of Southeast Asia (Diptera: Culicidae). Genetica 57:81-86. (14) Mahon RJ and PM Miethke. 1982. Anopheles farauti No. 3, a hitherto un

Life-table studies revealed significant effects of deforestation on the development and survivorship of Anopheles minimus larvae.

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Wang, Xiaoming; Zhou, Guofa; Zhong, Daibin; Wang, Xiaoling; Wang, Ying; Yang, Zhaoqing; Cui, Liwang; Yan, Guiyun

2016-06-06

Many developing countries are experiencing rapid ecological changes such as deforestation and shifting agricultural practices. These environmental changes may have an important consequence on malaria due to their impact on vector survival and reproduction. Despite intensive deforestation and malaria transmission in the China-Myanmar border area, the impact of deforestation on malaria vectors in the border area is unknown. We conducted life table studies on Anopheles minimus larvae to determine the pupation rate and development time in microcosms under deforested, banana plantation, and forested environments. The pupation rate of An. minimus was 3.8 % in the forested environment. It was significantly increased to 12.5 % in banana plantations and to 52.5 % in the deforested area. Deforestation reduced larval-to-pupal development time by 1.9-3.3 days. Food supplementation to aquatic habitats in forested environments and banana plantations significantly increased larval survival rate to a similar level as in the deforested environment. Deforestation enhanced the survival and development of An. minimus larvae, a major malaria vector in the China-Myanmar border area. Experimental determination of the life table parameters on mosquito larvae under a variety of environmental conditions is valuable to model malaria transmission dynamics and impact by climate and environmental changes.

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.

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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

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

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Hidalgo K.

2015-12-01

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

Effect of larval crowding on mating competitiveness of Anopheles gambiae mosquitoes

NARCIS (Netherlands)

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

2005-01-01

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

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

International Nuclear Information System (INIS)

Barfi, E.

2015-07-01

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

Habitat Perkembangbiakan Spesifik Anopheles sp Di Tambang Emas Kura-Kura Banian (Perubahan Perilaku Anopheles sp

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Liestiana Indriyati, SKM

2016-12-01

Full Text Available The results of Health Research 2010, showed that malaria was an infectious disease that the sixth cause of death cause of the sixth of death in Indonesia. Kotabaru is a malaria endemic district, especially in Puskesmas Banian, Annual Parasite Incidence 241.19 in 2014 where the cases of malaria with occurring in illegal gold mines on the slopes of Banian Mountain. It’s needed to know the factors that affect the existence of malaria in these locations. This research was observational study doing by cross-sectional study and analyzed by descriptively. Research conducted at the Gold Mine Banian Buluh Kuning village, Sungai Durian subdistric, Kotabaru district, South Kalimantan Province in February and March, 2015. The activities were catching larva and adult mosquitoes, environmental and mosquito breeding habitats observation and Mass Blood Survey. It was found Anopheles maculatus and Anopheles leucosphyrus with very low densities in area with the number of malaria cases by 22.99%. It was found divers breeding habitats of Anopheles sp on a former gold wash, former drum and former plastic containers. This indicated a behavioral change of Anopheles breeding places because previous to this the Anopheles breed in puddles on the ground or direct contact with the ground.

The effect of metal pollution on the life history and insecticide resistance phenotype of the major malaria vector Anopheles arabiensis (Diptera: Culicidae.

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Shüné V Oliver

Full Text Available Metal exposure is one of the commonest anthropogenic pollutants mosquito larvae are exposed to, both in agricultural and urban settings. As members of the Anopheles gambiae complex, which contains several major malaria vector species including An. arabiensis, are increasingly adapting to polluted environments, this study examined the effects of larval metal exposure on various life history traits of epidemiological importance. Two laboratory strains of An. arabiensis, SENN (insecticide susceptible and SENN DDT (insecticide resistant, were reared in maximum acceptable toxicity concentrations, (MATC-the highest legally accepted concentration of cadmium chloride, lead nitrate and copper nitrate. Following these exposures, time to pupation, adult size and longevity were determined. Larvae reared in double the MATC were assessed for changes in malathion and deltamethrin tolerance, measured by lethal time bottle bioassay, as well as changes in detoxification enzyme activity. As defence against oxidative stress has previously been demonstrated to affect the expression of insecticide resistance, catalase, glutathione peroxidase and superoxide dismutase activity was assessed. The relative metal toxicity to metal naïve larvae was also assessed. SENN DDT larvae were more tolerant of metal pollution than SENN larvae. Pupation in SENN larvae was significantly reduced by metal exposure, while adult longevity was not affected. SENN DDT showed decreased adult size after larval metal exposure. Adult insecticide tolerance was increased after larval metal exposure, and this effect appeared to be mediated by increased β-esterase, cytochrome P450 and superoxide dismutase activity. These data suggest an enzyme-mediated positive link between tolerance to metal pollutants and insecticide resistance in adult mosquitoes. Furthermore, exposure of larvae to metal pollutants may have operational consequences under an insecticide-based vector control scenario by increasing

Shading by Napier grass reduces malaria vector larvae in natural habitats in western Kenya highlands

NARCIS (Netherlands)

Wamae, P.M.; Githeko, A.K.; Menya, D.M.; Takken, W.

2010-01-01

Increased human population in the Western Kenya highlands has led to reclamation of natural swamps resulting in the creation of habitats suitable for the breeding of Anopheles gambiae, the major malaria vector in the region. Here we report on a study to restore the reclaimed swamp and reverse its

Rickettsia Species in African Anopheles Mosquitoes

Science.gov (United States)

Socolovschi, Cristina; Pages, Frédéric; Ndiath, Mamadou O.; Ratmanov, Pavel; Raoult, Didier

2012-01-01

Background There is higher rate of R. felis infection among febrile patients than in healthy people in Sub-Saharan Africa, predominantly in the rainy season. Mosquitoes possess a high vectorial capacity and, because of their abundance and aggressiveness, likely play a role in rickettsial epidemiology. Methodology/Principal Findings Quantitative and traditional PCR assays specific for Rickettsia genes detected rickettsial DNA in 13 of 848 (1.5%) Anopheles mosquitoes collected from Côte d’Ivoire, Gabon, and Senegal. R. felis was detected in one An. gambiae molecular form S mosquito collected from Kahin, Côte d’Ivoire (1/77, 1.3%). Additionally, a new Rickettsia genotype was detected in five An. gambiae molecular form S mosquitoes collected from Côte d’Ivoire (5/77, 6.5%) and one mosquito from Libreville, Gabon (1/88, 1.1%), as well as six An. melas (6/67, 9%) mosquitoes collected from Port Gentil, Gabon. A sequence analysis of the gltA, ompB, ompA and sca4 genes indicated that this new Rickettsia sp. is closely related to R. felis. No rickettsial DNA was detected from An. funestus, An. arabiensis, or An. gambiae molecular form M mosquitoes. Additionally, a BLAST analysis of the gltA sequence from the new Rickettsia sp. resulted in a 99.71% sequence similarity to a species (JQ674485) previously detected in a blood sample of a Senegalese patient with a fever from the Bandafassi village, Kedougou region. Conclusion R. felis was detected for the first time in An. gambiae molecular form S, which represents the major African malaria vector. The discovery of R. felis, as well as a new Rickettsia species, in mosquitoes raises new issues with respect to African rickettsial epidemiology that need to be investigated, such as bacterial isolation, the degree of the vectorial capacity of mosquitoes, the animal reservoirs, and human pathogenicity. PMID:23118963

Nueva especie de Parathelohania (Microsporidia en larvas de Anopheles aquasalis (Diptera: Culicidae en Venezuela

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Frances Osborn

2002-12-01

Full Text Available Se describen los síntomas en larvas de A. aquasalis infectadas naturalmente con un microsporidio parasítico, y la ultraestructura de las esporas del mismo. Las larvas fueron criadas bajo condiciones de laboratorio en concentraciones de salinidad de 10 g/l y 20 g/l, registrándose diariamente la mortalidad y la fecha del cambio de estadío de las mismas. Las larvas infectadas fueron procesadas por rnicroscopía electrónica de transmisión usando métodos convencionales con pH 7.2 y 260 mOsm/I. El incremento de la infección por los microsporidios estuvo correlacionado positivamente con un aumento en la duración promedio del cuarto estadío de 2.88 a 6.33 días en 10 g/l de sal y de 2.47 a 6.14 días en 20 g/l. La mortalidad de las larvas también aumentó en aproximadamente 50% durante este estadío a ambas concentraciones de salinidad. No se observaron estas alteraciones en los otros estadíos. Las esporas maduras de los microsporidios fueron encontradas en el intestino de las larvas. Estas esporas mostraron forma de barril y dimensiones de 2.6 x 2.4 µm aproximadamente. La reducción en la supervivencia de las larvas de A. aquasalis infectadas con los microsporidios, y el aumento del tiempo de su desarrollo sugiere que este parásito podría ser un posible controlador biológico de esta plaga. El microsporidio descrito presenta características similares a las del género Parathelohania. Se sugiere que el microsporidio encontrado en las larvas de A. aquasalis es una especie nueva y se propone nombrarla Parathelohania aquasalensis. Este trabajo representa el primer reporte de un microsporidio encontrado en un díptero en Venezuela.The symptoms of Anopheles aquasalis larvae naturally infected by a microsporidium, and the ultrastructure of the infecting spores is describes. The larvae were maintained under laboratory conditions in salt concentrations of 10 g/l and 20 g/l of water. Daily recordings of the mortality of the larvae were made and

Genomic, Physiologic, and Symbiotic Characterization of Serratia marcescens Strains Isolated from the Mosquito Anopheles stephensi

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Shicheng Chen

2017-08-01

Full Text Available Strains of Serratia marcescens, originally isolated from the gut lumen of adult female Anopheles stephensi mosquitoes, established persistent infection at high rates in adult A. stephensi whether fed to larvae or in the sugar meal to adults. By contrast, the congener S. fonticola originating from Aedes triseriatus had lower infection in A. stephensi, suggesting co-adaptation of Serratia strains in different species of host mosquitoes. Coinfection at high infection rate in adult A. stephensi resulted after feeding S. marcescens and Elizabethkingia anophelis in the sugar meal, but when fed together to larvae, infection rates with E. anophelis were much higher than were S. marcescens in adult A. stephensi, suggesting a suppression effect of coinfection across life stages. A primary isolate of S. marcescens was resistant to all tested antibiotics, showed high survival in the mosquito gut, and produced alpha-hemolysins which contributed to lysis of erythrocytes ingested with the blood meal. Genomes of two primary isolates from A. stephensi, designated S. marcescens ano1 and ano2, were sequenced and compared to other Serratia symbionts associated with insects, nematodes and plants. Serratia marcescens ano1 and ano2 had predicted virulence factors possibly involved in attacking parasites and/or causing opportunistic infection in mosquito hosts. S. marcescens ano1 and ano2 possessed multiple mechanisms for antagonism against other microorganisms, including production of bacteriocins and multi-antibiotic resistance determinants. These genes contributing to potential anti-malaria activity including serralysins, hemolysins and chitinases are only found in some Serratia species. It is interesting that genome sequences in S. marcescens ano1 and ano2 are distinctly different from those in Serratia sp. Ag1 and Ag2 which were isolated from Anopheles gambiae. Compared to Serratia sp. Ag1 and Ag2, S. marcescens ano1 and ano2 have more rRNAs and many important

Genomic, Physiologic, and Symbiotic Characterization of Serratia marcescens Strains Isolated from the Mosquito Anopheles stephensi.

Science.gov (United States)

Chen, Shicheng; Blom, Jochen; Walker, Edward D

2017-01-01

Strains of Serratia marcescens , originally isolated from the gut lumen of adult female Anopheles stephensi mosquitoes, established persistent infection at high rates in adult A. stephensi whether fed to larvae or in the sugar meal to adults. By contrast, the congener S. fonticola originating from Aedes triseriatus had lower infection in A. stephensi , suggesting co-adaptation of Serratia strains in different species of host mosquitoes. Coinfection at high infection rate in adult A. stephensi resulted after feeding S. marcescens and Elizabethkingia anophelis in the sugar meal, but when fed together to larvae, infection rates with E. anophelis were much higher than were S. marcescens in adult A. stephensi , suggesting a suppression effect of coinfection across life stages. A primary isolate of S. marcescens was resistant to all tested antibiotics, showed high survival in the mosquito gut, and produced alpha-hemolysins which contributed to lysis of erythrocytes ingested with the blood meal. Genomes of two primary isolates from A. stephensi , designated S. marcescens ano1 and ano2, were sequenced and compared to other Serratia symbionts associated with insects, nematodes and plants. Serratia marcescens ano1 and ano2 had predicted virulence factors possibly involved in attacking parasites and/or causing opportunistic infection in mosquito hosts. S. marcescens ano1 and ano2 possessed multiple mechanisms for antagonism against other microorganisms, including production of bacteriocins and multi-antibiotic resistance determinants. These genes contributing to potential anti-malaria activity including serralysins, hemolysins and chitinases are only found in some Serratia species. It is interesting that genome sequences in S. marcescens ano1 and ano2 are distinctly different from those in Serratia sp. Ag1 and Ag2 which were isolated from Anopheles gambiae . Compared to Serratia sp. Ag1 and Ag2, S. marcescens ano1 and ano2 have more rRNAs and many important genes involved in

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

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Knols Bart GJ

2010-06-01

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

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

Science.gov (United States)

2010-01-01

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

Larvicidal properties of three plants on Anopheles gambiae ...

African Journals Online (AJOL)

No larvae exposed to Q. africana developed into pupae while development was also curtailed when exposed to K. senegalensis and L. multiflora extracts. Larvicidal activity was concentration dependent. These plants could be sources of botanical insecticides for malarial control. Journal of Pharmacy and Bioresources, Vol.

Survivorship Of Anopheles gambiae In Relation To Malaria Transmission In Ilorin, Nigeria

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Israel Kayode Olayemi

2008-11-01

Full Text Available For the first time in Africa, an entomological study went beyond the conventional practice of determining parity and survival rates of field-collected adult anopheline mosquitoes but also related these variables to duration of Plasmodium sporogony and estimated the expectation of infective life. Blood-seeking female mosquitoes were collected in Ilorin, Nigeria, from January 2005 to December 2006, and dissected for ovarian tracheations following WHO recommended techniques. The results indicated an annual mean parous rate of 70.92%, and significantly higher parous rates in the rainy than dry season, which also had very low densities. Mean probability of daily survival of the mosquitoes was 0.80, with annual mean life expectancy of 12.24 days. The probability of surviving the sporogonic cycle was low (< 0.4 but the expectation of infective life was long, especially in the rainy season (mean = 8.31 days. The epidemiological implications of these results were discussed. The An. gambiae population in Ilorin is dominated by older mosquitoes with high survival rates thus, suggesting a high vector potential for the species in the area. These information on the survival rates of An. gambiae in relation to malaria transmission would enhance the development of a more focused and informed vector control interventions

Application of a qPCR assay in the investigation of susceptibility to malaria infection of the M and S molecular forms of An. gambiae s.s. in Cameroon.

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Anne Boissière

Full Text Available Plasmodium falciparum is the causative agent of malaria, a disease that kills almost one million persons each year, mainly in sub-Saharan Africa. P. falciparum is transmitted to the human host by the bite of an Anopheles female mosquito, and Anopheles gambiae sensus stricto is the most tremendous malaria vector in Africa, widespread throughout the afro-tropical belt. An. gambiae s.s. is subdivided into two distinct molecular forms, namely M and S forms. The two molecular forms are morphologically identical but they are distinct genetically, and differ by their distribution and their ecological preferences. The epidemiological importance of the two molecular forms in malaria transmission has been poorly investigated so far and gave distinct results in different areas. We have developed a real-time quantitative PCR (qPCR assay, and used it to detect P. falciparum at the oocyst stage in wild An. gambiae s.s. mosquitoes experimentally infected with natural isolates of parasites. Mosquitoes were collected at immature stages in sympatric and allopatric breeding sites and further infected at the adult stage. We next measured the infection prevalence and intensity in female mosquitoes using the qPCR assay and correlated the infection success with the mosquito molecular forms. Our results revealed different prevalence of infection between the M and S molecular forms of An. gambiae s.s. in Cameroon, for both sympatric and allopatric populations of mosquitoes. However, no difference in the infection intensity was observed. Thus, the distribution of the molecular forms of An. gambiae s.s. may impact on the malaria epidemiology, and it will be important to monitor the efficiency of malaria control interventions on the two M and S forms.

Contemporary evolution of resistance at the major insecticide target site gene Ace-1 by mutation and copy number variation in the malaria mosquito Anopheles gambiae

Science.gov (United States)

Weetman, David; Mitchell, Sara N; Wilding, Craig S; Birks, Daniel P; Yawson, Alexander E; Essandoh, John; Mawejje, Henry D; Djogbenou, Luc S; Steen, Keith; Rippon, Emily J; Clarkson, Christopher S; Field, Stuart G; Rigden, Daniel J; Donnelly, Martin J

2015-01-01

Functionally constrained genes are ideal insecticide targets because disruption is often fatal, and resistance mutations are typically costly. Synaptic acetylcholinesterase (AChE) is an essential neurotransmission enzyme targeted by insecticides used increasingly in malaria control. In Anopheles and Culex mosquitoes, a glycine–serine substitution at codon 119 of the Ace-1 gene confers both resistance and fitness costs, especially for 119S/S homozygotes. G119S in Anopheles gambiae from Accra (Ghana) is strongly associated with resistance, and, despite expectations of cost, resistant 119S alleles are increasing significantly in frequency. Sequencing of Accra females detected only a single Ace-1 119S haplotype, whereas 119G diversity was high overall but very low at non-synonymous sites, evidence of strong purifying selection driven by functional constraint. Flanking microsatellites showed reduced diversity, elevated linkage disequilibrium and high differentiation of 119S, relative to 119G homozygotes across up to two megabases of the genome. Yet these signals of selection were inconsistent and sometimes weak tens of kilobases from Ace-1. This unexpected finding is attributable to apparently ubiquitous amplification of 119S alleles as part of a large copy number variant (CNV) far exceeding the size of the Ace-1 gene, whereas 119G alleles were unduplicated. Ace-1 CNV was detectable in archived samples collected when the 119S allele was rare in Ghana. Multicopy amplification of resistant alleles has not been observed previously and is likely to underpin the recent increase in 119S frequency. The large CNV compromised localization of the strong selective sweep around Ace-1, emphasizing the need to integrate CNV analysis into genome scans for selection. PMID:25865270

Study on Fungal Flora in the Midgut of the Larva and Adult of the Different Populations of the Malaria Vector Anopheles stephensi

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L Tajedin

2009-07-01

Full Text Available Abstract Background: Many microorganisms in midgut of mosquito challenge with their host and also other pathogens pre­sent in midgut. The aim of this study was presence of non-pathogens microorganisms like fungal flora which may be cru­cial on interaction between vectors and pathogens."nMethods: Different populations of Anopheles stephensi were reared in insectary and objected to determine fungal flora in their midguts. The midgut paunch of mosquito adults and larvae as well as breading water and larval food sam­ples transferred on Subaru-dextrose agar, in order to detect the environment fungus."nResults: Although four fungi, Aspergillus, Rhizopus, Geotrichum and Sacharomyces were found in the food and wa­ter, but only Aspiragilus observed in the midgut of larvae. No fungus was found in the midgut of adults. This is the first report on fungal flora in the midgut of the adults and larvae of An. stephensi and possible stadial transmission of fungi from immature stages to adults."nConclusion: The midgut environment of adults is not compatible for survivorship of fungi but the larval midgut may con­tain few fungi as a host or even pathogen.

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

Science.gov (United States)

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

2014-01-01

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

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

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Mukabana Wolfgang R

2002-12-01

Full Text Available Abstract 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.

FAUNA DAN TEMPAT PERKEMBANGBIAKAN POTENSIAL NYAMUK Anopheles spp DI KECAMATAN MAYONG, KABUPATEN JEPARA, JAWA TENGAH

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Mardiana Mardiana

2012-10-01

Full Text Available Malaria masih merupakan masalah kesehatan masyarakat di beberapa daerah pedesaan di Jawa Tengah. Usaha pemberantasan malaria telah dilakukan oleh program baik secara kimiawi maupun hayati, guna memutuskan rantai penularan. Penelitian fauna dan tempat perindukan potensial nyamuk Anopheles telah dilakukan di Desa Buaran, Kecamatan Mayong I, Kabupaten Jepara, Jawa Tengah. Penangkapan nyamuk dengan umpan orang dilakukan di dalam dan di luar rumah pada malam hari dari pukul 18.00-24.00 yang masing-masing dilakukan oleh dua orang kolektor. Penangkapan nyamuk yang istirahat di dalam dan luar rumah (vegetasi pada pagi hari dilakukan pukul 06.00-08.00, yang dilakukan satu bulan 4 kali penangkapan selama 6 bulan. Pengambilan larva dan pupa dilakukan dari pukul 06.00-08.00 pagi di tempat genangan air dan sawah serta tempat yang potensial diduga sebagai perindukan Anopheles. Hasil penangkapan selama 6 bulan, diperoleh 1248 ekor nyamuk Anopheles yang terdiri dari 6 spesies yaitu: An. aconitus 442 ekor (35,42%, An. annularis 69 ekor (5,53% , An. barbirostris 30 ekor (2,4%, An. maculatus 2 ekor (0,16%, An. tesselatus 5 ekor (0,40% dan An. vagus 700 ekor (56,09%. Populasi aconitus ditemukan dari penangkapan di luar rumah, pada bulan Juli (56,40%, Agustus (42,80% dan Oktober (39,50% sedangkan pada bulan Mei (52,9%, Juni (44% dan September (50,40% dari penangkapan di kandang sapi. Pengambilan larva dan pupa Anopheles dilakukan di tempat habitat seperti sawah yang pada bulan Aguslus terbanyak ditemukan sebesar 85 (1.70, di sungai ditemukan hanya 4 (0.08 serta di genangan air bekas telapak kaki/kobokan ditemukan sebesar 6 (0.12. Ternyata tempat perindukan yang potensial larva Anopheles pada musim kemarau, ditemukan pada sungai yang ditanami kangkung oleh masyarakat selempat. Kata kunci: Fauna, tempat perindukan, Anopheles, vector

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

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Stone Chris M

2012-01-01

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

Effect of Bacillus sphaericus Neide on Anopheles (Diptera: Culicidae and associated insect fauna in fish ponds in the Amazon

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Francisco Augusto da Silva Ferreira

2015-09-01

Full Text Available ABSTRACTWe analyzed the effects of Bacillus sphaericus on Anopheles larvae and on the associated insect fauna in fish farming ponds. Five breeding sites in the peri-urban area of the city of Manaus, AM, Brazil, were studied. Seven samples were collected from each breeding site and B. sphaericus was applied and reapplied after 15 days. The samples were made at 24 h before application, 24 h post-application and 5 and 15 days post-application. We determined abundance, larval reduction and larval density for Anopheles, and abundance, richness, Shannon diversity index and classified according to the functional trophic groups for associated insect fauna. A total of 904 Anopheles larvae were collected and distributed into five species. Density data and larval reduction demonstrated the rapid effect of the biolarvicide 24 h after application. A total of 4874 associated aquatic insects belonging to six orders and 23 families were collected. Regression analysis of diversity and richness indicated that the application of the biolarvicide had no influence on these indices and thus no effect on the associated insect fauna for a period of 30 days. B. sphaericus was found to be highly effective against the larvae of Anopheles, eliminating the larvae in the first days after application, with no effect on the associated insect fauna present in the fish ponds analyzed.

Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strategies for malaria vector control in Africa using attractive toxic sugar bait methods

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Bah Sekou

2010-09-01

Full Text Available Abstract Background Based on recent studies in Israel demonstrating that attractive toxic sugar bait (ATSB methods can be used to decimate local anopheline and culicine mosquito populations, an important consideration is whether the same methods can be adapted and improved to attract and kill malaria vectors in Africa. The ATSB approach uses fruit or flower scent as an attractant, sugar solution as a feeding stimulant, and an oral toxin. The ATSB solutions are either sprayed on vegetation or suspended in simple bait stations, and the mosquitoes ingesting the toxic solutions are killed. As such, this approach targets sugar-feeding female and male mosquitoes. This study examines the attractiveness of African malaria vectors to local fruits/seedpods and flowering plants, key biological elements of the ATSB approach for mosquito control. Methods Three field experiments were conducted at sites in Mali. The attraction of Anopheles gambiae s.l. to 26 different local fruits and seedpods was determined at a site in the semi-arid Bandiagara District of Mali. Wire mesh glue traps with fruits/seedpods suspended on skewers inside were set along a seasonal lagoon. Seven replicates of each fruit/seedpod species were tested, with a water-soaked sponge and a sugar-soaked sponge as controls. The attraction of An. gambiae s.l. to 26 different types of flowering plants was determined at a site near Mopti in Mali. The flowering plants held in a water-filled buried container were tested using the same glue traps, with controls including water only and sugar solution. Six replicates of each selected plant type were tested on transects between rice paddies. Additional studies using CDC light traps were done to determine the relative densities and periodicity of An. gambiae s.l. attraction to branches of the most highly attractive flowering plant, branches without flowers, human odor, and candescent light. Results Of the 26 fruits and seedpods tested, 6 were attractive

Field experiments of Anopheles gambiae attraction to local fruits/seedpods and flowering plants in Mali to optimize strategies for malaria vector control in Africa using attractive toxic sugar bait methods.

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Müller, Günter C; Beier, John C; Traore, Sekou F; Toure, Mahamoudou B; Traore, Mohamed M; Bah, Sekou; Doumbia, Seydou; Schlein, Yosef

2010-09-20

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. 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. Of the 26 fruits and seedpods tested, 6 were attractive to An. gambiae s.l. females and males, respectively

Microsporidia parásitos de larvas de mosquito de la Costa Pacífica del Chocó

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Zuluaga Juan S.

1993-12-01

Full Text Available Two genera of Microsporidia were found infecting mosquito larvae in three localities on the Pacific coast of Choco. Vavraia sp. (Microsporida: Pleistophoridae was found in larvae of Wyeomyia circumcincta, W. simmsi and Anopheles albimanus collected from plants of the Bromeliacea family in Arusí y Joví. Amblyospora sp. (Microsporida: Amblyosporidae was found parasitizingAedes angustivittatuslarvae COllectedfrom a terrestrial breeding pond in the locality of Nuqur. Morphology of the spores of the two parasites under light microscopy is described, as well as preliminary data on host range when exposed to laboratory rearad Aedes aegypti, Culex quinquefasciatus and Anopheles albimanus. Their rola in mosquito control is discussed.Se reportan dos géneros de microsporidia que parasitan larvas de mosquitos en criaderos naturales de tres localidades en la costa Pacffica Chocoana. Vavraia sp. (Microsporida: Pleistophoridae parásita larvas de Wyeomyia circumcincta, de Wyeomya simmsi y de Anopheles neivai, recolectadas en las rosetas de especies de la familia Bromeliaceae en las localidades de Arusí y Joví. Amblyospora sp. (Microsporida: Amblyosporidae parásita larvas deAedes angustivittatusde criaderos terrestres semipermanentes en la localidad de Nuquí. Se describe la morfología de estos dos microsporidia al microscopio óptico. Estudios preliminares de infección en larvas de Aedes aegypti, Culex quinquefasciatus yAnopheles albimanus, criadas en laboratorio, indican que Vavraia sp. infecta las tres especies, con preferencia a Culex quinquefasciatus. Las larvas expuestas a esporas de Amblyospora sp. no presentaron infección. Se discute el posible papel de estos dos géneros en el control de las poblaciones de mosquitos.

Mosquito host preferences affect their response to synthetic and natural odour blends

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Busula, A.O.; Takken, W.; Loy, D.E.; Hahn, B.H.; Mukabana, W.R.; Verhulst, N.O.

2015-01-01

Background The anthropophilic malaria mosquito Anopheles gambiae sensu stricto (hereafter termed Anopheles gambiae) primarily takes blood meals from humans, whereas its close sibling Anopheles arabiensis is more opportunistic. Previous studies have identified several compounds that play a critical

Microbial Pre-exposure and Vectorial Competence of Anopheles Mosquitoes

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Constentin Dieme

2017-12-01

Full Text Available Anopheles female mosquitoes can transmit Plasmodium, the malaria parasite. During their aquatic life, wild Anopheles mosquito larvae are exposed to a huge diversity of microbes present in their breeding sites. Later, adult females often take successive blood meals that might also carry different micro-organisms, including parasites, bacteria, and viruses. Therefore, prior to Plasmodium ingestion, the mosquito biology could be modulated at different life stages by a suite of microbes present in larval breeding sites, as well as in the adult environment. In this article, we highlight several naturally relevant scenarios of Anopheles microbial pre-exposure that we assume might impact mosquito vectorial competence for the malaria parasite: (i larval microbial exposures; (ii protist co-infections; (iii virus co-infections; and (iv pathogenic bacteria co-infections. In addition, significant behavioral changes in African Anopheles vectors have been associated with increasing insecticide resistance. We discuss how these ethological modifications may also increase the repertoire of microbes to which mosquitoes could be exposed, and that might also influence their vectorial competence. Studying Plasmodium–Anopheles interactions in natural microbial environments would efficiently contribute to refining the transmission risks.

Morphology of the larvae, male genitalia and DNA sequences of Anopheles (Kerteszia pholidotus (Diptera: Culicidae from Colombia

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Jesús Eduardo Escovar

2014-07-01

Full Text Available Since 1984, Anopheles (Kerteszia lepidotus has been considered a mosquito species that is involved in the transmission of malaria in Colombia, after having been incriminated as such with epidemiological evidence from a malaria outbreak in Cunday-Villarrica, Tolima. Subsequent morphological analyses of females captured in the same place and at the time of the outbreak showed that the species responsible for the transmission was not An. lepidotus, but rather Anopheles pholidotus. However, the associated morphological stages and DNA sequences of An. pholidotus from the foci of Cunday-Villarrica had not been analysed. Using samples that were caught recently from the outbreak region, the purpose of this study was to provide updated and additional information by analysing the morphology of female mosquitoes, the genitalia of male mosquitoes and fourth instar larvae of An. pholidotus, which was confirmed with DNA sequences of cytochrome oxidase I and rDNA internal transcribed spacer. A total of 1,596 adult females were collected in addition to 37 larval collections in bromeliads. Furthermore, 141 adult females, which were captured from the same area in the years 1981-1982, were analysed morphologically. Ninety-five DNA sequences were analysed for this study. Morphological and molecular analyses showed that the species present in this region corresponds to An. pholidotus. Given the absence of An. lepidotus, even in recent years, we consider that the species of mosquitoes that was previously incriminated as the malaria vector during the outbreak was indeed An. pholidotus, thus ending the controversy.

Low prevalence of Plasmodium and absence of malaria transmission in Conakry, Guinea: prospects for elimination.

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Kouassi, Bernard L; de Souza, Dziedzom K; Goepogui, Andre; Balde, Siradiou M; Diakité, Lamia; Sagno, Arsène; Djameh, Georgina I; Chammartin, Frédérique; Vounatsou, Penelope; Bockarie, Moses J; Utzinger, Jürg; Koudou, Benjamin G

2016-03-18

Over the past 15 years, mortality and morbidity due to malaria have been reduced substantially in sub-Saharan Africa and local elimination has been achieved in some settings. This study addresses the bio-ecology of larval and adult stages of malaria vectors, Plasmodium infection in Anopheles gambiae s.l. in the city of Conakry, Guinea, and discusses the prospect for malaria elimination. Water bodies were prospected to identify potential mosquito breeding sites for 6 days each in the dry season (January 2013) and in the rainy season (August 2013), using the dipping method. Adult mosquitoes were collected in 15 communities in the five districts of Conakry using exit traps and indoor spraying catches over a 1-year period (November 2012 to October 2013). Molecular approaches were employed for identification of Anopheles species, including An. coluzzii and An. gambiae s.s. Individual An. gambiae mosquitoes were tested for Plasmodium falciparum and P. vivax sporozoites using the VecTest™ malaria panel assay and an enzyme-linked immunosorbent assay. A systematic research of Ministry of Health statistical yearbooks was performed to determine malaria prevalence in children below the age of 5 years. Culex larval breeding sites were observed in large numbers throughout Conakry in both seasons. While Anopheles larval breeding sites were less frequent than Culex breeding sites, there was a high odds of finding An. gambiae mosquito larvae in agricultural sites during the rainy season. Over the 1-year study period, a total of 14,334 adult mosquitoes were collected; 14,135 Culex (98.6%) and 161 (1.1%) from the An. gambiae complex. One-hundred and twelve Anopheles mosquitoes, mainly collected from rice fields and gardens, were subjected to molecular analysis. Most of the mosquitoes were An. gambiae s.s. (n = 102; 91.1%) while the remaining 10 (8.9%) were An. melas. The molecular M form of An. gambiae s.s. was predominant (n = 89; 79.5%). The proportions of kdr genotype in the An

Assessment of Clarias gariepinus as a biological control agent against mosquito larvae.

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Chala, Buze; Erko, Berhanu; Animut, Abebe; Degarege, Abraham; Petros, Beyene

2016-05-31

The emergence and spread of insecticide resistant mosquitoes renewed interest in investigating the use of larvivorous fish as a biological control agent. The potential of Clarias gariepinus fish in controlling Anopheles arabiensis and culicine larvae was assessed under laboratory and semi-field conditions. Small size (15-20 cm) C. gariepinus fish consumed greater number of mosquito larvae than the large size fish (25-40 cm) in the multivariate regression model (β = 13.36, 95 % CI = 4.57, 22.15). The Anopheles larvae consumed was greater in number than the culicines larvae consumed by the fish (β = 12.10, 95 % CI = 3.31, 20.89). The number of larvae consumed was greater during the night hours than during the light hours (β = 30.06, 95 % CI = 21.27, 38.85). Amount of supplementary fish food did not cause significant differences in the number of mosquito larvae consumed by the fish among different groups. C. gariepinus was observed to feed on mosquito larvae under laboratory and semi-field conditions. C. gariepinus fed on the larvae of An. arabiensis and culicines readily. Hence, it can be used as an alternative mosquito control agent in Ethiopia where the breeding habitats are small and localized.

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

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Sheila B Ogoma

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

Species composition, co-occurrence, association and affinity indices of mosquito larvae (Diptera: Culicidae) in Mazandaran Province, northern Iran.

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Nikookar, Seyed Hassan; Azari-Hamidian, Shahyad; Fazeli-Dinan, Mahmoud; Nasab, Seyed Nouraddin Mousavi; Aarabi, Mohsen; Ziapour, Seyyed Payman; Enayati, Ahmadali

2016-05-01

Although considerable progress has been made in the past years in management of mosquito borne diseases such as malaria, dengue, yellow fever and West Nile fever through research in biology and ecology of the vectors, these diseases are still major threats to human health. Therefore, more research is required for better management of the diseases. This investigation provides information on the composition, co-occurrence, association and affinity indices of mosquito larvae in Mazandaran Province, northern Iran. In a large scale field study, mosquito larvae were collected from 120 sentinel sites in 16 counties in Mazandaran Province, using standard 350 ml dipper. Sampling took place monthly from May to December 2014. Collected larvae were mounted on glass slides using de Faure's medium and were diagnosed using morphological characters. Totally, 19,840 larvae were collected including three genera and 16 species from 120 larval habitats, as follows: Anopheles claviger, Anopheles hyrcanus, Anopheles maculipennis s.l., Anopheles marteri, Anopheles plumbeus, Anopheles pseudopictus, Culex pipiens, Culex tritaeniorhynchus, Culex torrentium, Culex perexiguus, Culex territans, Culex mimeticus, Culex hortensis, Culiseta annulata, Culiseta longiareolata, and Culiseta morsitans. Predominant species were Cx. pipiens and An. maculipennis s.l. which show the highest co-occurrence. The pair of species An. hyrcanus/An. pseudopictus showed significant affinity and association. High co-occurrence of the predominant species Cx. pipiens and An. maculipennis s.l. in the study area is of considerable importance in terms of vector ecology. It was also revealed that An. pseudopictus/An. hyrcanus often occur sympatrically indicating their common habitat requirements. The information may be equally important when vector control measures are considered. Copyright © 2016 Elsevier B.V. All rights reserved.

A de novo expression profiling of Anopheles funestus, malaria vector in Africa, using 454 pyrosequencing.

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Richard Gregory

2011-02-01

Full Text Available Anopheles funestus is one of the major malaria vectors in Africa and yet there are few genomic tools available for this species compared to An. gambiae. To start to close this knowledge gap, we sequenced the An. funestus transcriptome using cDNA libraries developed from a pyrethroid resistant laboratory strain and a pyrethroid susceptible field strain from Mali.Using a pool of life stages (pupae, larvae, adults: females and males for each strain, 454 sequencing generated 375,619 reads (average length of 182 bp. De novo assembly generated 18,103 contigs with average length of 253 bp. The average depth of coverage of these contigs was 8.3. In total 20.8% of all reads were novel when compared to reference databases. The sequencing of the field strain generated 204,758 reads compared to 170,861 from the insecticide resistant laboratory strain. The contigs most differentially represented in the resistant strain belong to the P450 gene family and cuticular genes which correlates with previous studies implicating both of these gene families in pyrethroid resistance. qPCR carried out on six contigs indicates that these ESTs could be suitable for gene expression studies such as microarray. 31,000 sites were estimated to contain Single Nucleotide Polymorphisms (SNPs and analysis of SNPs from 20 contigs suggested that most of these SNPs are likely to be true SNPs. Gene conservation analysis confirmed the close phylogenetic relationship between An. funestus and An. gambiae.This study represents a significant advance for the genetics and genomics of An. funestus since it provides an extensive set of both Expressed Sequence Tags (ESTs and SNPs which can be readily adopted for the design of new genomic tools such as microarray or SNP platforms.

Response of Anopheles gambiae detoxification enzymes to levels of ...

African Journals Online (AJOL)

Some of the sampled larvae were reared to pupae and adult life stages. Levels of 7 physical (pH, temperature, conductivity, transparency, total dissolved solids, dissolved oxygen and biological oxygen demand) and 6 chemical (sulphates, phosphates, nitrites, nitrates, carbon content and oil and grease) environmental ...

Anopheles (Nyssorhynchus deaneorum: a new species in the albitarsis complex (Diptera: culicidae Anopheles (Nyssorhynchus deaneorum: uma nova espécie no complexo albitarsis (Diptera: Culicidae

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Maria Goreti Rosa-Freitas

1989-12-01

Full Text Available Anopheles (Nyssorhynchus deaneorum sp. n. is described from specimens collected in Guajará-Mirim, Rondônia state and Rio Branco, Acre state, Brazil, on human and animal baits, inside dwellings and from the progenies of engorged females. A detailed description of the shape of egg, external appearance of adult female and male, genitalias, female cibarial armature and complete chaetotaxy of pupa and larva show that it can be distinguished from Anopheles albitarsis from the type-locality and other areas by the paler general external appearance of the adult, the posterolateral tufts of scales, on the female abdominal terga and the branching of the outer anterior clypeal seta (3-C of the fourth instar larva (as shown in illustrations. If species can also be distinguished from An. albitarsis from the type locality by the allele frequencies at 11 enzymic loci as represented by Nei's Genetic Distance.Anopheles (Nyssorhynchus deaneorum sp. n. é descrito a partir de exemplares coletados em capturas comparativas no intradomicílio e no peridomicílio usando isca humana e animal e progênies de fêmeas ingurgitadas, em Guarajá-Mirim, Rondônia e Rio Branco, Acre. A descrição detalhada do ovo, dos adultos fêmea e macho, inclusive cibário da fêmea, genitália, quetotaxia da pupa e da larva e seu perfil isoenzimático, mostram que esse mosquito pode ser distinguido do Anopheles albitarsis na fase adulta pelo aspecto geral mais claro, pela presença de tufos laterais de escamas escuras somente a partir do quarto ou quinto tergitos abdominais, enquanto em albitarsis começam no terceiro e, na fase larvária, pela ramificação das cerdas clipeais anteriores externas, que em albitarsis são aciculadas (como mostram as ilustrações, bem como pelo padrão isoenzimático.

Contemporary evolution of resistance at the major insecticide target site gene Ace-1 by mutation and copy number variation in the malaria mosquito Anopheles gambiae.

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Weetman, David; Mitchell, Sara N; Wilding, Craig S; Birks, Daniel P; Yawson, Alexander E; Essandoh, John; Mawejje, Henry D; Djogbenou, Luc S; Steen, Keith; Rippon, Emily J; Clarkson, Christopher S; Field, Stuart G; Rigden, Daniel J; Donnelly, Martin J

2015-06-01

Functionally constrained genes are ideal insecticide targets because disruption is often fatal, and resistance mutations are typically costly. Synaptic acetylcholinesterase (AChE) is an essential neurotransmission enzyme targeted by insecticides used increasingly in malaria control. In Anopheles and Culex mosquitoes, a glycine-serine substitution at codon 119 of the Ace-1 gene confers both resistance and fitness costs, especially for 119S/S homozygotes. G119S in Anopheles gambiae from Accra (Ghana) is strongly associated with resistance, and, despite expectations of cost, resistant 119S alleles are increasing significantly in frequency. Sequencing of Accra females detected only a single Ace-1 119S haplotype, whereas 119G diversity was high overall but very low at non-synonymous sites, evidence of strong purifying selection driven by functional constraint. Flanking microsatellites showed reduced diversity, elevated linkage disequilibrium and high differentiation of 119S, relative to 119G homozygotes across up to two megabases of the genome. Yet these signals of selection were inconsistent and sometimes weak tens of kilobases from Ace-1. This unexpected finding is attributable to apparently ubiquitous amplification of 119S alleles as part of a large copy number variant (CNV) far exceeding the size of the Ace-1 gene, whereas 119G alleles were unduplicated. Ace-1 CNV was detectable in archived samples collected when the 119S allele was rare in Ghana. Multicopy amplification of resistant alleles has not been observed previously and is likely to underpin the recent increase in 119S frequency. The large CNV compromised localization of the strong selective sweep around Ace-1, emphasizing the need to integrate CNV analysis into genome scans for selection. © 2015 The Authors. Molecular Ecology published by John Wiley & Sons Ltd.

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

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Braks, M.

1999-01-01

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

The dominant Anopheles vectors of human malaria in Africa, Europe and the Middle East: occurrence data, distribution maps and bionomic précis

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Okara Robi M

2010-12-01

Full Text Available Abstract Background This is the second in a series of three articles documenting the geographical distribution of 41 dominant vector species (DVS of human malaria. The first paper addressed the DVS of the Americas and the third will consider those of the Asian Pacific Region. Here, the DVS of Africa, Europe and the Middle East are discussed. The continent of Africa experiences the bulk of the global malaria burden due in part to the presence of the An. gambiae complex. Anopheles gambiae is one of four DVS within the An. gambiae complex, the others being An. arabiensis and the coastal An. merus and An. melas. There are a further three, highly anthropophilic DVS in Africa, An. funestus, An. moucheti and An. nili. Conversely, across Europe and the Middle East, malaria transmission is low and frequently absent, despite the presence of six DVS. To help control malaria in Africa and the Middle East, or to identify the risk of its re-emergence in Europe, the contemporary distribution and bionomics of the relevant DVS are needed. Results A contemporary database of occurrence data, compiled from the formal literature and other relevant resources, resulted in the collation of information for seven DVS from 44 countries in Africa containing 4234 geo-referenced, independent sites. In Europe and the Middle East, six DVS were identified from 2784 geo-referenced sites across 49 countries. These occurrence data were combined with expert opinion ranges and a suite of environmental and climatic variables of relevance to anopheline ecology to produce predictive distribution maps using the Boosted Regression Tree (BRT method. Conclusions The predicted geographic extent for the following DVS (or species/suspected species complex* is provided for Africa: Anopheles (Cellia arabiensis, An. (Cel. funestus*, An. (Cel. gambiae, An. (Cel. melas, An. (Cel. merus, An. (Cel. moucheti and An. (Cel. nili*, and in the European and Middle Eastern Region: An. (Anopheles atroparvus

BIONOMIK VEKTOR MALARIA NYAMUK Anopheles sundaicus dan Anopheles letifer DI KECAMATAN BELAKANG PADANG , BATAM, KEPULAUAN RIAU

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Shinta Shinta

2012-11-01

Full Text Available Abstract Malaria continues to be a public health problem in the malaria endemic areas in Indonesia and often cause an outbreak. Batam municipality is the priority for development area in the Riau island Province, nevertheless malaria is still a public health problem. The national government and district office government have been committed to have a program for eliminating malaria at Batam area in year 2015. One of the malaria control program is the vector control measure. The failure of vector control is partly due to a lack of understanding of vector behavior in its epidemiological setting. The understanding of  malaria vector species and its behavior will be useful to plan the vector control intervention.  The study of bio-ecology of malaria vector is very important factor to  understand its behavior and to formulate the vector control strtegy in Batam area. This study was carried out at Belakang Padang, Batam in 2008 using breeding habitat survey of Anopheles spp, measuring the pH, salinity and observation of breeding characteristics, mapping of breeding sites distribution using GPS and human landing collection inside as well as outside houses and ELISA for circumsporozoite.  The results of the study revealed that, in the Belakang padang areas were found five natural breeding habitat of Anopheles spp. e.i: marshy areas, marshy with mangrove tree in the peripher, creek, mud-hole and water reservoir (water dam. Larvae of  An. letifer and An sundaicus were found relatively higher number in the marshy areas, with characteristis of pH: 5-7,5,  temperature 28-330C and salinity was 0-28 ‰.  Only one species of anopheline An. sundaicus  was found in the adult stage in Belakang Padang area, though An. letifer was found only in the larva stage.  The biting activities of An. sundaicus throughout the night both indoors and outdoors, though the biting peak occured at 02.00-03.00 am.  Both  An sundaicus and An. letifer at  Belakang Padang

Highly focused anopheline breeding sites and malaria transmission in Dakar

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Bouzid Samia

2009-06-01

Full Text Available Abstract Background Urbanization has a great impact on the composition of the vector system and malaria transmission dynamics. In Dakar, some malaria cases are autochthonous but parasite rates and incidences of clinical malaria attacks have been recorded at low levels. Ecological heterogeneity of malaria transmission was investigated in Dakar, in order to characterize the Anopheles breeding sites in the city and to study the dynamics of larval density and adult aggressiveness in ten characteristically different urban areas. Methods Ten study areas were sampled in Dakar and Pikine. Mosquitoes were collected by human landing collection during four nights in each area (120 person-nights. The Plasmodium falciparum circumsporozoite (CSP index was measured by ELISA and the entomological inoculation rates (EIR were calculated. Open water collections in the study areas were monitored weekly for physico-chemical characterization and the presence of anopheline larvae. Adult mosquitoes and hatched larvae were identified morphologically and by molecular methods. Results In September-October 2007, 19,451 adult mosquitoes were caught among which, 1,101 were Anopheles gambiae s.l. The Human Biting Rate ranged from 0.1 bites per person per night in Yoff Village to 43.7 in Almadies. Seven out of 1,101 An. gambiae s.l. were found to be positive for P. falciparum (CSP index = 0.64%. EIR ranged from 0 infected bites per person per year in Yoff Village to 16.8 in Almadies. The An. gambiae complex population was composed of Anopheles arabiensis (94.8% and Anopheles melas (5.2%. None of the An. melas were infected with P. falciparum. Of the 54 water collection sites monitored, 33 (61.1% served as anopheline breeding sites on at least one observation. No An. melas was identified among the larval samples. Some physico-chemical characteristics of water bodies were associated with the presence/absence of anopheline larvae and with larval density. A very close parallel

UJI COBA LARVISIDA SPHERIFIX (Bacillus sphaericus VCRC B 42 TERHADAP LARVA Anopheles sundaicus Di GERUMBUL KLACES, UJUNG ALANG - KABUPATEN CILACAP

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Umi Widyastuti

2012-09-01

Full Text Available A biological larvicide spherifix containing Bacillus sphaericus VCRC B 42 was investigated against Anopheles sundaicus in Klaces hamlet, Cilacap regency. This study was conducted to determine the effectivity of spherifix on An. sundaicus larvae at a dosage of 2.5 kg/Ha. Observations were conducted one day before application of the larvicide, 24, 36, 48 hours, day 4, 7, and 14 after application. The larval reduction rates were calculated using the formula of Mulia et al, 1971, and a reduction of the results were 16.69 % after 24 hours, 20.95 % after 36 hours, 34.07 % after 48 hours, 65.08 % after 4 days, 85.98 % after 7 days, and 90.81 % after 14 days. B. sphaericus has capabilities to function as a biological larvicide.

Toxicity of (Burseraceae Leaf Fractions against Immature Stages of (Giles and (Say (Diptera: Culicidae

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Lame Younoussa

2016-01-01

Full Text Available 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 (LC 50 = 385.9 ppm and C. quinquefasciatus (LC 50 = 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 .

Genome-Wide Divergence in the West-African Malaria Vector Anopheles melas

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Kevin C. Deitz

2016-09-01

Full Text Available Anopheles melas is a member of the recently diverged An. gambiae species complex, a model for speciation studies, and is a locally important malaria vector along the West-African coast where it breeds in brackish water. A recent population genetic study of An. melas revealed species-level genetic differentiation between three population clusters. An. melas West extends from The Gambia to the village of Tiko, Cameroon. The other mainland cluster, An. melas South, extends from the southern Cameroonian village of Ipono to Angola. Bioko Island, Equatorial Guinea An. melas populations are genetically isolated from mainland populations. To examine how genetic differentiation between these An. melas forms is distributed across their genomes, we conducted a genome-wide analysis of genetic differentiation and selection using whole genome sequencing data of pooled individuals (Pool-seq from a representative population of each cluster. The An. melas forms exhibit high levels of genetic differentiation throughout their genomes, including the presence of numerous fixed differences between clusters. Although the level of divergence between the clusters is on a par with that of other species within the An. gambiae complex, patterns of genome-wide divergence and diversity do not provide evidence for the presence of pre- and/or postmating isolating mechanisms in the form of speciation islands. These results are consistent with an allopatric divergence process with little or no introgression.

Efficacy of local neem extracts for sustainable malaria vector control in an African village

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Duchemin Jean-Bernard

2008-07-01

Full Text Available Abstract Background Larval control of malaria vectors has been historically successful in reducing malaria transmission, but largely fell out of favour with the introduction of synthetic insecticides and bed nets. However, an integrated approach to malaria control, including larval control methods, continues to be the best chance for success, in view of insecticide resistance, the behavioural adaptation of the vectors to changing environments and the difficulties of reaching the poorest populations most at risk,. Laboratory studies investigating the effects of neem seed (Azadirachta indica extracts on Anopheles larvae have shown high rates of larval mortality and reductions in adult longevity, as well as low potential for resistance development. Methods This paper describes a method whereby seeds of the neem tree can be used to reduce adult Anopheles gambiae s.l. abundance in a way that is low cost and can be implemented by residents of rural villages in western Niger. The study was conducted in Banizoumbou village, western Niger. Neem seeds were collected from around the village. Dried seeds were ground into a coarse powder, which was then sprinkled onto known Anopheles larvae breeding habitats twice weekly during the rainy season 2007. Adult mosquitoes were captured on a weekly basis in the village and captures compared to those from 2005 and 2006 over the same period. Adult mosquitoes were also captured in a nearby village, Zindarou, as a control data set and compared to those from Banizoumbou. Results It was found that twice-weekly applications of the powder to known breeding habitats of Anopheles larvae in 2007 resulted in 49% fewer adult female Anopheles gambiae s.l. mosquitoes in Banizoumbou, compared with previous captures under similar environmental conditions and with similar habitat characteristics in 2005 and 2006. The productivity of the system in 2007 was found to be suppressed compared to the mean behaviour of 2005 and 2006 in

Wild Anopheles funestus mosquito genotypes are permissive for infection with the rodent malaria parasite, Plasmodium berghei.

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Jiannong Xu

Full Text Available Malaria parasites undergo complex developmental transitions within the mosquito vector. A commonly used laboratory model for studies of mosquito-malaria interaction is the rodent parasite, P. berghei. Anopheles funestus is a major malaria vector in sub-Saharan Africa but has received less attention than the sympatric species, Anopheles gambiae. The imminent completion of the A. funestus genome sequence will provide currently lacking molecular tools to describe malaria parasite interactions in this mosquito, but previous reports suggested that A. funestus is not permissive for P. berghei development.An A. funestus population was generated in the laboratory by capturing female wild mosquitoes in Mali, allowing them to oviposit, and rearing the eggs to adults. These F1 progeny of wild mosquitoes were allowed to feed on mice infected with a fluorescent P. berghei strain. Fluorescence microscopy was used to track parasite development inside the mosquito, salivary gland sporozoites were tested for infectivity to mice, and parasite development in A. funestus was compared to A. gambiae.P. berghei oocysts were detectable on A. funestus midguts by 7 days post-infection. By 18-20 days post-infection, sporozoites had invaded the median and distal lateral lobes of the salivary glands, and hemocoel sporozoites were observed in the hemolymph. Mosquitoes were capable of infecting mice via bite, demonstrating that A. funestus supports the complete life cycle of P. berghei. In a random sample of wild mosquito genotypes, A. funestus prevalence of infection and the characteristics of parasite development were similar to that observed in A. gambiae-P. berghei infections.The data presented in this study establish an experimental laboratory model for Plasmodium infection of A. funestus, an important vector of human malaria. Studying A. funestus-Plasmodium interactions is now feasible in a laboratory setting. This information lays the groundwork for exploitation of the

A simple Chelex protocol for DNA extraction from Anopheles spp.

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Musapa, Mulenga; Kumwenda, Taida; Mkulama, Mtawa; Chishimba, Sandra; Norris, Douglas E; Thuma, Philip E; Mharakurwa, Sungano

2013-01-09

Endemic countries are increasingly adopting molecular tools for efficient typing, identification and surveillance against malaria parasites and vector mosquitoes, as an integral part of their control programs. For sustainable establishment of these accurate approaches in operations research to strengthen malaria control and elimination efforts, simple and affordable methods, with parsimonious reagent and equipment requirements are essential. Here we present a simple Chelex-based technique for extracting malaria parasite and vector DNA from field collected mosquito specimens. We morphologically identified 72 Anopheles gambiae sl. from 156 mosquitoes captured by pyrethrum spray catches in sleeping rooms of households within a 2,000 km(2) vicinity of the Malaria Institute at Macha. After dissection to separate the head and thorax from the abdomen for all 72 Anopheles gambiae sl. mosquitoes, the two sections were individually placed in 1.5 ml microcentrifuge tubes and submerged in 20 μl of deionized water. Using a sterile pipette tip, each mosquito section was separately homogenized to a uniform suspension in the deionized water. Of the ensuing homogenate from each mosquito section, 10 μl was retained while the other 10 μl was transferred to a separate autoclaved 1.5 ml tube. The separate aliquots were subjected to DNA extraction by either the simplified Chelex or the standard salting out extraction protocol(9,10). The salting out protocol is so-called and widely used because it employs high salt concentrations in lieu of hazardous organic solvents (such as phenol and chloroform) for the protein precipitation step during DNA extraction(9). Extracts were used as templates for PCR amplification using primers targeting arthropod mitochondrial nicotinamide adenine dinucleotide dehydrogenase (NADH) subunit 4 gene (ND4) to check DNA quality, a PCR for identification of Anopheles gambiae sibling species(10) and a nested PCR for typing of Plasmodium falciparum infection

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

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

2013-08-01

Blocking transmission of malaria is a reliable way to control and eliminate infection. However, in-depth knowledge of the interaction between Plasmodium and mosquito is needed. Studies suggest that innate immunity is the main mechanism inhibiting development of malaria parasites in the mosquito. Recent studies have found that use of antibiotics that inhibit the mosquito gut flora can reduce the immune response of Anopheles gambiae, thereby contributing to the development of malaria parasites. In our study, we used the non susceptible model of Anopheles dirus-Plasmodium yoelii to explore the effect of Anopheles intestinal flora on the natural resistance of A. dirus to P. yoelii. We found that in mosquitoes infected with Plasmodium, the intestinal flora can regulate expression of thioester-containing protein (TEP1) via an RNAi gene-silencing approach. Our results suggest that in the absence of TEP1, the natural microbiota cannot suppress the development of P. yoelii in A. dirus. This suggests that AdTEP1 plays an important role in the resistance of A. dirus to P. yoelii. The intestinal flora may modulate the development of P. yoelii in A. dirus by regulating TEP1 expression. Copyright © 2013 Elsevier Inc. All rights reserved.

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

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Sedofia, B. K

2013-07-01

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

Experimental hut evaluation of bednets treated with an organophosphate (chlorpyrifos-methyl or a pyrethroid (lambdacyhalothrin alone and in combination against insecticide-resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes

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Corbel Vincent

2005-05-01

Full Text Available Abstract Background Pyrethroid resistant mosquitoes are becoming increasingly common in parts of Africa. It is important to identify alternative insecticides which, if necessary, could be used to replace or supplement the pyrethroids for use on treated nets. Certain compounds of an earlier generation of insecticides, the organophosphates may have potential as net treatments. Methods Comparative studies of chlorpyrifos-methyl (CM, an organophosphate with low mammalian toxicity, and lambdacyhalothrin (L, a pyrethroid, were conducted in experimental huts in Côte d'Ivoire, West Africa. Anopheles gambiae and Culex quinquefasciatus mosquitoes from the area are resistant to pyrethroids and organophosphates (kdr and insensitive acetylcholinesterase Ace.1R. Several treatments and application rates on intact or holed nets were evaluated, including single treatments, mixtures, and differential wall/ceiling treatments. Results and Conclusion All of the treatments were effective in reducing blood feeding from sleepers under the nets and in killing both species of mosquito, despite the presence of the kdr and Ace.1R genes at high frequency. In most cases, the effects of the various treatments did not differ significantly. Five washes of the nets in soap solution did not reduce the impact of the insecticides on A. gambiae mortality, but did lead to an increase in blood feeding. The three combinations performed no differently from the single insecticide treatments, but the low dose mixture performed encouragingly well indicating that such combinations might be used for controlling insecticide resistant mosquitoes. Mortality of mosquitoes that carried both Ace.1R and Ace.1S genes did not differ significantly from mosquitoes that carried only Ace.1S genes on any of the treated nets, indicating that the Ace.1R allele does not confer effective resistance to chlorpyrifos-methyl under the realistic conditions of an experimental hut.

The invasive shrub Prosopis juliflora enhances the malaria parasite transmission capacity of Anopheles mosquitoes: a habitat manipulation experiment.

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Muller, Gunter C; Junnila, Amy; Traore, Mohamad M; Traore, Sekou F; Doumbia, Seydou; Sissoko, Fatoumata; Dembele, Seydou M; Schlein, Yosef; Arheart, Kristopher L; Revay, Edita E; Kravchenko, Vasiliy D; Witt, Arne; Beier, John C

2017-07-05

A neglected aspect of alien invasive plant species is their influence on mosquito vector ecology and malaria transmission. Invasive plants that are highly attractive to Anopheles mosquitoes provide them with sugar that is critical to their survival. The effect on Anopheles mosquito populations was examined through a habitat manipulation experiment that removed the flowering branches of highly attractive Prosopis juliflora from selected villages in Mali, West Africa. Nine villages in the Bandiagara district of Mali were selected, six with flowering Prosopis juliflora, and three without. CDC-UV light traps were used to monitor their Anopheles spp. vector populations, and recorded their species composition, population size, age structure, and sugar feeding status. After 8 days, all of the flowering branches were removed from three villages and trap catches were analysed again. Villages where flowering branches of the invasive shrub Prosopis juliflora were removed experienced a threefold drop in the older more dangerous Anopheles females. Population density dropped by 69.4% and the species composition shifted from being a mix of three species of the Anopheles gambiae complex to one dominated by Anopheles coluzzii. The proportion of sugar fed females dropped from 73 to 15% and males from 77 to 10%. This study demonstrates how an invasive plant shrub promotes the malaria parasite transmission capacity of African malaria vector mosquitoes. Proper management of invasive plants could potentially reduce mosquito populations and malaria transmission.

Genome-wide identification and characterization of odorant-binding protein (OBP) genes in the malaria vector Anopheles sinensis (Diptera: Culicidae).

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He, Xiu; He, Zheng-Bo; Zhang, Yu-Juan; Zhou, Yong; Xian, Peng-Jie; Qiao, Liang; Chen, Bin

2016-06-01

Anopheles sinensis is a major malaria vector. Insect odorant-binding proteins (OBPs) may function in the reception of odorants in the olfactory system. The classification and characterization of the An. sinensis OBP genes have not been systematically studied. In this study, 64 putative OBP genes were identified at the whole-genome level of An. sinensis based on the comparison between OBP conserved motifs, PBP_GOBP, and phylogenetic analysis with An. gambiae OBPs. The characterization of An. sinensis OBPs, including the motif's conservation, gene structure, genomic organization and classification, were investigated. A new gene, AsOBP73, belonging to the Plus-C subfamily, was identified with the support of transcript and conservative motifs. These An. sinensis OBP genes were classified into three subfamilies with 37, 15 and 12 genes in the subfamily Classic, Atypical and Plus-C, respectively. The genomic organization of An. sinensis OBPs suggests a clustered distribution across nine different scaffolds. Eight genes (OBP23-28, OBP63-64) might originate from a single gene through a series of historic duplication events at least before divergence of Anopheles, Culex and Aedes. The microsynteny analyses indicate a very high synteny between An. sinensis and An. gambiae OBPs. OBP70 and OBP71 earlier classified under Plus-C in An. gambiae are recognized as belonging to the group Obp59a of the Classic subfamily, and OBP69 earlier classified under Plus-C has been moved to the Atypical subfamily in this study. The study established a basic information frame for further study of the OBP genes in insects as well as in An. sinensis. © 2016 Institute of Zoology, Chinese Academy of Sciences.

Activity of Ocimum basilicum, Ocimum canum, and Cymbopogon citratus essential oils against Plasmodium falciparum and mature-stage larvae of Anopheles funestus s.s.

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Akono Ntonga Patrick

2014-01-01

Full Text Available The biological activities of essential oils from three plants grown in Cameroon: Ocimum basilicum, Ocimum canum, and Cymbopogon citratus were tested against Plasmodium falciparum and mature-stage larvae of Anopheles funestus. Gas chromatography and gas chromatography – mass spectrometry analyses showed that the main compounds are geranial, 1,8-cineole and linalool in C. citratus, O. canum and O. basilicum, respectively. Larvicidal tests carried out according to the protocol recommended by the World Health Organization showed that the essential oil of leaves of C. citratus is the most active against larvae of An. funestus (LC50 values = 35.5 ppm and 34.6 ppm, respectively, for larval stages III and IV after 6 h of exposure. Besides, the in vitro anti-plasmodial activity evaluated by the radioisotopic method showed that the C. citratus oil is the most active against P. falciparum, with an IC50 value of 4.2 ± 0.5 μg/mL compared with O. canum (20.6 ± 3.4 μg/mL and O. basilicum (21 ± 4.6 μg/mL. These essential oils can be recommended for the development of natural biocides for fighting the larvae of malaria vectors and for the isolation of natural products with anti-malarial activity.

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

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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.

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

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Antonio-Nkondjio, Christophe; Fossog, Billy Tene; Ndo, Cyrille; Djantio, Benjamin Menze; Togouet, Serge Zebaze; Awono-Ambene, Parfait; Costantini, Carlo; Wondji, Charles S; Ranson, Hilary

2011-06-08

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

Impact of PermaNet 3.0 on entomological indices in an area of pyrethroid resistant Anopheles gambiae in south-western Nigeria

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2014-01-01

Background PermaNet® 3.0 is an insecticide synergist-combination long-lasting insecticidal net designed to have increased efficacy against malaria vectors with metabolic resistance, even when combined with kdr. The current study reports on the impact of this improved tool on entomological indices in an area with pyrethroid-resistant malaria vectors in Nigeria. Methods Baseline entomological indices across eight villages in Remo North LGA of Ogun State provided the basis for selection of three villages (Ilara, Irolu and Ijesa) for comparing the efficacy of PermaNet® 3.0 (PN3.0), PermaNet® 2.0 (PN2.0) and untreated polyester nets as a control (UTC). In each case, nets were distributed to cover all sleeping spaces and were evaluated for insecticidal activity on a 3-monthly basis. Collection of mosquitoes was conducted monthly via window traps and indoor resting catches. The arithmetic means of mosquito catches per house, entomological inoculation rates before and during the intervention were compared as well as three other outcome parameters: the mean mosquito blood feeding rate, mean mortality and mean parity rates. Results Anopheles gambiae s.l. was the main malaria vector in the three villages, accounting for >98% of the Anopheles population and found in appreciable numbers for 6–7 months. Deltamethrin, permethrin and lambdacyhalothrin resistance were confirmed at Ilara, Irolu and Ijesa. The kdr mutation was the sole resistance mechanism at Ilara, whereas kdr plus P450-based metabolic mechanisms were detected at Irolu and Ijesa. Bioassays repeated on domestically used PN 2.0 and PN 3.0 showed persistent optimal (100%) bio-efficacy for both net types after the 3rd, 6th, 9th and 12th month following net distribution. The use of PN 3.0 significantly reduced mosquito densities with a ‘mass killing’ effect inside houses. Households with PN 3.0 also showed reduced blood feeding as well as lower mosquito parity and sporozoite rates compared to the PN 2.0 and the

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

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Ball Amanda

2007-08-01

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

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

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Bass, Chris; Nikou, Dimitra; Donnelly, Martin J; Williamson, Martin S; Ranson, Hilary; Ball, Amanda; Vontas, John; Field, Linda M

2007-01-01

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

Sebaran Nyamuk Anopheles pada Topografi Wilayah yang Berbeda di Provinsi Jambi

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Yulian Taviv

2015-09-01

Full Text Available Penularan penyakit tular vektor seperti malaria dipengaruhi oleh banyak faktor. Salah satu faktor yangtelah diketahui memiliki asosiasi dengan malaria adalah topograf wilayah yang erat hubungannya denganpola penularan. Berdasarkan tempat atau lokasi terhadap penyakit yang ditularkan oleh vektor makaperlu diperhatikan pembagian zoogeografi dimana jenis-jenis nyamuk di setiap lokasi akan dipengaruhifaktor-faktor lingkungan di setiap daerah yang berbeda. Penelitian ini bertujuan untuk mengetahui jenisAnopheles serta habitat perkembangbiakannya pada dua wilayah dengan topograf yang berbeda diProvinsi Jambi. Kegiatan yang dilakukan adalah penangkapan nyamuk dewasa dengan metode humanlanding collection dan survei habitat perkembangbiakan Anopheles. Penangkapan nyamuk dilakukanselama 12 jam dimulai dari jam 18.00 WIB hingga jam 06.00 WIB. Larva Anopheles yang berhasilditangkap selanjutnya dibawa ke laboratorium dan dipelihara hingga dewasa dan selanjutnya diidentifiasijenisnya. Hasil penangkapan nyamuk Anopheles di Desa Nipah Panjang Kabupaten Tanjung JabungTimur (dataran rendah adalah An. separatus, An. sinensis, An. tesselatus dan An. letifer. Anophelesletifer memiliki angka tertinggi untuk nilai kekerapan 3,33, kelimpahan nisbi 40, dominansi 133,33 danMan Bitting Rate (MBR 0,07. Penangkapan nyamuk Anopheles di Desa Teluk Rendak KabupatenSarolangun (dataran tinggi meliputi An. nigerrimus, An. annularis, An. letifer, An. maculatus dan An.barbumbrosus. Anopheles nigerrimus memiliki angka tertinggi untuk nilai kekerapan 21,67, kelimpahannisbi 60,98, dominansi 1321,14 dan MBR 0,63.

Mitochondrial genomes of Anopheles arabiensis, An.gambiae and An.coluzzii show no clear species division

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Here we report the complete mitochondrial sequences of 70 individual field collected mosquito specimens from throughout Sub-Saharan Africa. We generated this dataset to identify species specific markers for the following Anopheles species and chromosomal forms: An.arabiensis, An.coluzzii (The Forest...

BIONOMIK NYAMUK MANSONIA DAN ANOPHELES DI DESA KARYA MAKMUR, KABUPATEN OKU TIMUR

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Yanelza Supranelfy

2016-09-01

Full Text Available Abstract Lymphatic filariasis is still become the health problem in Indonesia, the disease almost found inthe entire area with fairly high level endemicity. Karya Makmur Village, Madang Suku III District, EastOgan Komering Ulu Regency, South Sumatera Province in 2007 has micofilariae rate of 1,05% based fromblood survey. The aims of the research is to identify vector and the breeding habitat. This research has beenconducted in Karya Makmur Village from May to November 2011. This type of research is noninterventionstudy with spot survey research design. Mosquitoes collection using human landing andresting collection methods, while mosquito larvae were collected from breeding sites. This research found14 species of Mansonia and Anopheles were collected and the species were Mansonia uniformis, Ma.annulata, Ma. indiana, Ma. dives, Ma. bonneae, Ma. annulifera, Anopheles nigerrimus , An. annularis, An.maculatus, An. letifer, An. vagus, An. barbumbrosus, An. barbirostris and An. tesselatus. Mansoniauniformis and Anopheles nigerrimus was confirmed as filariasis vector but none containing microfilariae.Breeding habitat of Anopheles have water temperature 280C — 300C and pH 7 with vegetation (grass andEichhornia crassipes / water hyacinth and predator (Oreochromis niloticus / nile tilapia and Aplocheiluspanchax / fish head lead. Thus the community are expected to behave positively, especially avoidmosquito bites and manipulate the environment that potentially as mosquito larvae breeding habitat.Keywords: Mansonia, Anopheles, filariasis, Karya Makmur Village AbstrakFilariasis limfatik masih merupakan masalah kesehatandi Indonesia, penyebaran penyakit ini hampirdi seluruh wilayah dan dibeberapa daerah dengan tingkatendemic yang cukup tinggi. Desa Karya Makmur, Kecamatan Madang SukuIII, Kabupaten Ogan KomeringUlu Timur, Propinsi Sumatera Selatan pada tahun2007, mikrofilariarate sebesar1,05% berdasarkansurvey darah jari. Tujuan penelitian untuk mengetahui

Laboratory Evaluations of the Fractions Efficacy of Annona senegalensis (Annonaceae) Leaf Extract on Immature Stage Development of Malarial and Filarial Mosquito Vectors.

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Lame, Younoussa; Nukenine, Elias Nchiwan; Pierre, Danga Yinyang Simon; Elijah, Ajaegbu Eze; Esimone, Charles Okechukwu

2015-12-01

Within the framework to control mosquitoes, ovicidal, larvicidal and pupicidal activity of Annona senegalensis leaf extract and its 4 fractions against Anopheles gambiae and Culex quinquefasciatus were evaluated in the laboratory conditions. Ovicidal test was performed by submitting at least 100 eggs of mosquitoes to 125, 250, 500, 1000 and 2000 ppm concentrations, while larvicidal and pupicidal effects were assessed by submitting 25 larvae or pupae to the concentrations of 2500, 1250, 625 and 312.5 ppm of plant extract or fractions of A. senegalensis. The eggs of An. gambiae were most affected by N-hexane (0.00% hatchability) and chloroform (03.67% hatchability) fractions compared to Cx. quinquefasciatus where at least 25 % hatchability were recorded at 2000 ppm. For larvicidal test, N-hexane (LC50= 298.8 ppm) and chloroform (LC50= 418.3 ppm) fractions were more effective than other fractions on An. gambiae larvae while, a moderate effectiveness was also observed with N-hexane (LC50= 2087.6 ppm), chloroform (LC50= 9010.1 ppm) fractions on Cx. quinquefasciatus larvae. The highest mortality percent of the pupae were also recorded with N-hexane and chloroform fractions on An. gambiae at 2500 ppm. As for Cx. quinquefasciatus only 50 % and 36 % mortality were recorded with N-hexane and chloroform fractions respectively. The extract of A. senegalensis was toxic on immature stage of mosquito species tested. By splitting methanolic crude extract, only N-hexane and chloroform fractions were revealed to possess a mosquitocidal effects and could be considered and utilized for future immature mosquito vectors control.

Laboratory Evaluations of the Fractions Efficacy of Annona senegalensis (Annonaceae Leaf Extract on Immature Stage Development of Malarial and Filarial Mosquito Vectors

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Younoussa Lame

2015-10-01

Full Text Available Background: Within the framework to control mosquitoes, ovicidal, larvicidal and pupicidal activity of Annona senegalensis leaf extract and its 4 fractions against Anopheles gambiae and Culex quinquefasciatus were evaluated in the laboratory conditions.Methods: Ovicidal test was performed by submitting at least 100 eggs of mosquitoes to 125, 250, 500, 1000 and 2000 ppm concentrations, while larvicidal and pupicidal effects were assessed by submitting 25 larvae or pupae to the concentrations of 2500, 1250, 625 and 312.5 ppm of plant extract or fractions of A. senegalensis.Results: The eggs of An. gambiae were most affected by N-hexane (0.00% hatchability and chloroform (03.67% hatchability fractions compared to Cx. quinquefasciatus where at least 25 % hatchability were recorded at 2000 ppm. For larvicidal test, N-hexane (LC50= 298.8 ppm and chloroform (LC50= 418.3 ppm fractions were more effective than other fractions on An. gambiae larvae while, a moderate effectiveness was also observed with Nhexane (LC50= 2087.6 ppm, chloroform (LC50= 9010.1 ppm fractions on Cx. quinquefasciatus larvae. The highest mortality percent of the pupae were also recorded with N-hexane and chloroform fractions on An. gambiae at 2500 ppm. As for Cx. quinquefasciatus only 50 % and 36 % mortality were recorded with N-hexane and chloroform fractions respectively.Conclusion: The extract of A. senegalensis was toxic on immature stage of mosquito species tested. By splitting methanolic crude extract, only N-hexane and chloroform fractions were revealed to possess a mosquitocidal effects and could be considered and utilized for future immature mosquito vectors control.

Identification of the temperature- induced larvicidal efficacy of Agave angustifolia against Aedes, Culex and Anopheles larvae

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Mithilesh eKajla

2016-01-01

Full Text Available Synthetic insecticides are generally employed to control the mosquito population. However, their injudicious over usage and non-biodegradability are associated with many adverse effects on the environment and mosquitoes. The application of environment-friendly mosquitocidals might be an alternate to overcome these issues. In this study, we found that organic or aqueous extracts of Agave angustifolia leaves exhibited a strong larvicidal activity (LD50 28.27 µg/ml against Aedes aegypti, Culex quinquefasciatus and Anopheles stephensi larvae within a short exposure of 12h. The larvicidal activity of Agave angustifolia is inherited and independent of the plants vegetative growth. Interestingly, the plant larvicidal activity was observed exclusively during the summer season (April-August, when outside temperature is between 30oC to 50oC and it was significantly reduced during winter season (December-February, when the outside temperature falls to ~4oC or lower. Thus, we hypothesized that the larvicidal components of Agave angustifolia might be induced by the manipulation of environmental temperature and should be resistant to the hot conditions. We found that the larvicidal activity of Agave angustifolia was induced when plants were maintained at 37oC in a semi-natural environment against the controls that were growing outside in cold weather. Pre-incubation of Agave angustifolia extract at 100oC for 1h killed 60% larvae in 12h, which gradually increased to 100% mortality after 24h. In addition, the dry powder formulation of Agave angustifolia, also displayed a strong larvicidal activity after a long shelf life. Together, these findings revealed that Agave angustifolia is an excellent source of temperature induced bioactive metabolites that may assist the preparedness for vector control programs competently.

Confirmation of Anopheles (Anopheles calderoni Wilkerson, 1991 (Diptera: Culicidae in Colombia and Ecuador through molecular and morphological correlation with topotypic material

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Ranulfo González

2010-12-01

Full Text Available The morphologically similar taxa Anopheles calderoni, Anopheles punctimacula, Anopheles malefactor and Anopheles guarao are commonly misidentified. Isofamilies collected in Valle de Cauca, Colombia, showed morphological characters most similar to An. calderoni, a species which has never previously been reported in Colombia. Although discontinuity of the postsubcostal pale spots on the costa (C and first radial (R1 wing veins is purportedly diagnostic for An. calderoni, the degree of overlap of the distal postsubcostal spot on C and R1 were variable in Colombian specimens (0.003-0.024. In addition, in 98.2% of larvae, seta 1-X was located off the saddle and seta 3-C had 4-7 branches in 86.7% of specimens examined. Correlation of DNA sequences of the second internal transcribed spacer and mtDNA cytochrome c oxidase subunit I gene (COI barcodes (658 bp of the COI gene generated from Colombian progeny material and wild-caught mosquitoes from Ecuador with those from the Peruvian type series of An. calderoni confirmed new country records. DNA barcodes generated for the closely related taxa, An. malefactor and An. punctimacula are also presented for the first time. Examination of museum specimens at the University of the Valle, Colombia, revealed the presence of An. calderoni in inland localities across Colombia and at elevations up to 1113 m.

Pyrethroid resistance in an Anopheles funestus population from Uganda.

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John C Morgan

2010-07-01

Full Text Available The susceptibility status of Anopheles funestus to insecticides remains largely unknown in most parts of Africa because of the difficulty in rearing field-caught mosquitoes of this malaria vector. Here we report the susceptibility status of the An. funestus population from Tororo district in Uganda and a preliminary characterisation of the putative resistance mechanisms involved.A new forced egg laying technique used in this study significantly increased the numbers of field-caught females laying eggs and generated more than 4000 F1 adults. WHO bioassays indicated that An. funestus in Tororo is resistant to pyrethroids (62% mortality after 1 h exposure to 0.75% permethrin and 28% mortality to 0.05% deltamethrin. Suspected DDT resistance was also observed with 82% mortality. However this population is fully susceptible to bendiocarb (carbamate, malathion (organophosphate and dieldrin with 100% mortality observed after exposure to each of these insecticides. Sequencing of a fragment of the sodium channel gene containing the 1014 codon conferring pyrethroid/DDT resistance in An. gambiae did not detect the L1014F kdr mutation but a correlation between haplotypes and resistance phenotype was observed indicating that mutations in other exons may be conferring the knockdown resistance in this species. Biochemical assays suggest that resistance in this population is mediated by metabolic resistance with elevated level of GSTs, P450s and pNPA compared to a susceptible strain of Anopheles gambiae. RT-PCR further confirmed the involvement of P450s with a 12-fold over-expression of CYP6P9b in the Tororo population compared to the fully susceptible laboratory colony FANG.This study represents the first report of pyrethroid/DDT resistance in An. funestus from East Africa. With resistance already reported in southern and West Africa, this indicates that resistance in An. funestus may be more widespread than previously assumed and therefore this should be taken

[Identification of anopheles breeding sites in the residual foci of low malaria transmission «hotspots» in Central and Western Senegal].

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Sy, O; Konaté, L; Ndiaye, A; Dia, I; Diallo, A; Taïrou, F; Bâ, E L; Gomis, J F; Ndiaye, J L; Cissé, B; Gaye, O; Faye, O

2016-02-01

Malaria incidence has markedly declined in the Mbour, Fatick, Niakhar and Bambey districts (central and western Senegal) thanks to a scaling up of effective control measures namely LLINs (Long Lasting Insecticide Treated Net), ACTs (Artesunate Combination Therapy) and promoting care seeking. However malaria cases are now maintained by foci of transmission called hotspots. We evaluate the role of anopheles breeding sites in the identification of malaria hotspots in the health districts of Mbour, Fatick, Niakhar and Bambey. Surveys of breeding sites were made in 6 hotspot villages and 4 non-hotspot villages. A sample was taken in each water point with mosquito larvae by dipping method and the collected specimens were identified to the genus level. Additional parameters as name of the village and breeding sites, type of collection, original water turbidity, presence of vegetation, proximity to dwellings, geographic coordinates, sizes were also collected. Sixty-two water collections were surveyed and monitored between 2013 and 2014. Temporary natural breeding sites were predominant regardless of the epidemiological status of the village. Among the 31 breeding sites located within 500 meters of dwellings in hotspots villages, 70% carried Anopheles larvae during the rainy season while 43% of the 21 breeding sites located at similar distances in non-hotspot villages carried Anopheles larvae during the same period (P = 0.042). At the end of the rainy season, the trend is the same with 27% of positive breeding sites in hotspots and 14% in non-hotspots villages. The breeding sites encountered in hotspots villages are mostly small to medium size and are more productive by Anopheles larvae than those found in non-hotspot area. This study showed that the high frequency of smallest and productive breeding sites around and inside the villages can create conditions of residual transmission.

Seasonal abundance and blood feeding activity of Anopheles minimus Theobald (Diptera: Culicidae) in Thailand.

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Chareonviriyaphap, Theeraphap; Prabaripai, Atchariya; Bangs, Michael J; Aum-Aung, Boonserm

2003-11-01

Anopheline mosquito larvae and adults were sampled at Ban Pu Teuy, Tri-Yok District, Kanchanaburi Province, western Thailand, from January 2000 to December 2001. Over the period of 2 yr, Anopheles minimus sensu lato was the most commonly collected species, followed by Anopheles swadiwongporni and Anopheles dirus sensu lato; all three species are important vectors of malaria in Thailand. Attempted blood feeding by An. minimus occurred throughout the night, with two distinct feeding peaks: strong activity immediately after sunset (1800-2100 hours), followed by a second, less pronounced, rise before sunrise (0300-0600 hours). Anopheles minimus were more abundant during the wet season compared with the dry and hot seasons, although nocturnal adult feeding patterns were similar. Anopheles minimus fed readily on humans inside and outside of houses, showing a slight preference for exophagy. The human-biting peak of An. minimus in our study area differed from other localities sampled in Thailand, indicating the possible existence of site-specific populations of An. minimus exhibiting different host-seeking behavior. These results underscore the importance of conducting site-specific studies to accurately determine vector larval habitats and adult activity patterns and linking their importance in malaria transmission in a given area.

Development of an allele-specific, loop-mediated, isothermal amplification method (AS-LAMP to detect the L1014F kdr-w mutation in Anopheles gambiae s. l.

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Badolo Athanase

2012-07-01

Full Text Available Abstract Background Malaria control relies heavily on treated bed nets and indoor residual spraying with pyrethroid insecticides. Unfortunately, the resistance to pyrethroid insecticides, mainly due to the kdr mutation, is spreading in the main malaria vector Anopheles gambiae s.l., decreasing the insecticides’ efficacy. To manage the insecticide resistance rapidly and flexibly, simple and effective tools for the early detection of resistant mosquitoes are needed. This study aimed to develop an allele-specific, loop-mediated, isothermal amplification (AS-LAMP method to detect the West African-type kdr mutation (kdr-w; L1014F in field-collected mosquitoes. Methods DNA fragments of the wild-type and the mutated kdr gene were used to select the primers and develop the method. The primers were designed with the mutation at the 5’ end of the backward inner primer (BIP. The AS-LAMP method was compared to the AS-PCR method using the genomic DNA of 120 field-collected mosquitoes. Results The AS-LAMP method could discriminate between the wild-type homozygote, the heterozygote, and the kdr-w homozygote within 75 min. The AS-LAMP method has the advantage of being faster and at least as sensitive and specific as the AS-PCR method. Conclusions The AS-LAMP method can be used to detect the kdr mutation for quick decision-making, even in less well-equipped laboratories.

Fauna of mosquito larvae (Diptera: Culicida) in Asir Provence, Kingdom of Saudi Arabia.

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Al Ashry, Hamdy A; Kenawy, Mohamed A; Shobrak, Mohammed

2014-04-01

An entomological survey was undertaken for one year to update the mosquito fauna of Asir Region, Kingdom of Saudi Arabia. A total of 31 species of 8 genera were reported of which genus Culex (55%) was the most common. Most of collected larvae (59%) belonged to genus Culex (+ Lutzia) followed by Culiseta (26%), Anopheles (13%) and Aedine spp. (2%). Cx. pipiens (39%) and Cs. longiareolata (26.%) were generally the most abundant of all collected larvae. Of the Anopheles spp., An. dthali was common (40%), of Culex spp., Cx. pipiens was predominating (66%) and of Aedine spp., St. aegypti was predominating (71%). Four species: An. fluviatilis, Cx. mattinglyi, Cx. arbieeni and Cx. mimeticus were new reports in Asir Region and Cx. wigglesworthi recorded for the first time from the kingdom. Larvae were more common in low- and highlands than in the moderately altitude areas. In general all species prefer stagnant water but with the exception of Aedine larvae (altogether), the other species prefer presence of algae, vegetation and shade and absence of turbidity (except Culex spp.). A total of 98 different forms of association were reported of which 9 forms were common. All genera breed year round with peaks of abundance during spring for Anopheles spp. and Culex spp. and during winter for Aedine spp. and Cs. longiareolata. A complete list of mosquito fauna of Asir Region comprising 45 spp. was presented based on the present and previous surveys. The study concluded that the occurrence and prevalence of mosquito species mainly the disease vectors in Asir carry the thread of maintaining and transmission of several mosquito-borne diseases.

Sequencing and analysis of the complete mitochondrial genome in Anopheles sinensis (Diptera: Culicidae).

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Chen, Kai; Wang, Yan; Li, Xiang-Yu; Peng, Heng; Ma, Ya-Jun

2017-10-02

Anopheles sinensis (Diptera: Culicidae) is a primary vector of Plasmodium vivax and Brugia malayi in most regions of China. In addition, its phylogenetic relationship with the cryptic species of the Hyrcanus Group is complex and remains unresolved. Mitochondrial genome sequences are widely used as molecular markers for phylogenetic studies of mosquito species complexes, of which mitochondrial genome data of An. sinensis is not available. An. sinensis samples was collected from Shandong, China, and identified by molecular marker. Genomic DNA was extracted, followed by the Illumina sequencing. Two complete mitochondrial genomes were assembled and annotated using the mitochondrial genome of An. gambiae as reference. The mitochondrial genomes sequences of the 28 known Anopheles species were aligned and reconstructed phylogenetic tree by Maximum Likelihood (ML) method. The length of complete mitochondrial genomes of An. sinensis was 15,076 bp and 15,138 bp, consisting of 13 protein-coding genes, 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and an AT-rich control region. As in other insects, most mitochondrial genes are encoded on the J strand, except for ND5, ND4, ND4L, ND1, two rRNA and eight tRNA genes, which are encoded on the N strand. The bootstrap value was set as 1000 in ML analyses. The topologies restored phylogenetic affinity within subfamily Anophelinae. The ML tree showed four major clades, corresponding to the subgenera Cellia, Anopheles, Nyssorhynchus and Kerteszia of the genus Anopheles. The complete mitochondrial genomes of An. sinensis were obtained. The number, order and transcription direction of An. sinensis mitochondrial genes were the same as in other species of family Culicidae.

Bacterial diversity analysis of larvae and adult midgut microflora using culture-dependent and culture-independent methods in lab-reared and field-collected Anopheles stephensi-an Asian malarial vector

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Adak Tridibesh

2009-05-01

Full Text Available Abstract Background Mosquitoes are intermediate hosts for numerous disease causing organisms. Vector control is one of the most investigated strategy for the suppression of mosquito-borne diseases. Anopheles stephensi is one of the vectors of malaria parasite Plasmodium vivax. The parasite undergoes major developmental and maturation steps within the mosquito midgut and little is known about Anopheles-associated midgut microbiota. Identification and characterization of the mosquito midgut flora is likely to contribute towards better understanding of mosquito biology including longevity, reproduction and mosquito-pathogen interactions that are important to evolve strategies for vector control mechanisms. Results Lab-reared and field-collected A. stephensi male, female and larvae were screened by "culture-dependent and culture-independent" methods. Five 16S rRNA gene library were constructed form lab and field-caught A. stephensi mosquitoes and a total of 115 culturable isolates from both samples were analyzed further. Altogether, 68 genera were identified from midgut of adult and larval A. stephensi, 53 from field-caught and 15 from lab-reared mosquitoes. A total of 171 and 44 distinct phylotypes having 85 to 99% similarity with the closest database matches were detected among field and lab-reared A. stephensi midgut, respectively. These OTUs had a Shannon diversity index value of 1.74–2.14 for lab-reared and in the range of 2.75–3.49 for field-caught A. stephensi mosquitoes. The high species evenness values of 0.93 to 0.99 in field-collected adult and larvae midgut flora indicated the vastness of microbial diversity retrieved by these approaches. The dominant bacteria in field-caught adult male A. stephensi were uncultured Paenibacillaceae while in female and in larvae it was Serratia marcescens, on the other hand in lab-reared mosquitoes, Serratia marcescens and Cryseobacterium meninqosepticum bacteria were found to be abundant. Conclusion

GNBP domain of Anopheles darlingi: are polymorphic inversions and gene variation related to adaptive evolution?

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Bridi, L C; Rafael, M S

2016-02-01

Anopheles darlingi is the main malaria vector in humans in South America. In the Amazon basin, it lives along the banks of rivers and lakes, which responds to the annual hydrological cycle (dry season and rainy season). In these breeding sites, the larvae of this mosquito feed on decomposing organic and microorganisms, which can be pathogenic and trigger the activation of innate immune system pathways, such as proteins Gram-negative binding protein (GNBP). Such environmental changes affect the occurrence of polymorphic inversions especially at the heterozygote frequency, which confer adaptative advantage compared to homozygous inversions. We mapped the GNBP probe to the An. darlingi 2Rd inversion by fluorescent in situ hybridization (FISH), which was a good indicator of the GNBP immune response related to the chromosomal polymorphic inversions and adaptative evolution. To better understand the evolutionary relations and time of divergence of the GNBP of An. darlingi, we compared it with nine other mosquito GNBPs. The results of the phylogenetic analysis of the GNBP sequence between the species of mosquitoes demonstrated three clades. Clade I and II included the GNBPB5 sequence, and clade III the sequence of GNBPB1. Most of these sequences of GNBP analyzed were homologous with that of subfamily B, including that of An. gambiae (87 %), therefore suggesting that GNBP of An. darling belongs to subfamily B. This work helps us understand the role of inversion polymorphism in evolution of An. darlingi.

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

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Mohamed Yacoob Syed Ali

2013-06-01

Full Text Available 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 LC 50 value (0.055 6依0.010 3 µg/mL, (0.067 5依0.136 0 µg/mL and (0.066 1 依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.

'A mate or a meal' – Pre-gravid behaviour of female Anopheles gambiae from the islands of São Tomé and Príncipe, West Africa

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Ferreira Conceicao

2003-05-01

Full Text Available Abstract Background Malaria prevalence differs between the two islands that comprise the archipelago of São Tomé and Príncipe. This may be due to differences in the biology of local Anopheles gambiae, the only vector on the islands. Survival rate and feeding frequency are two factors influencing vectorial capacity. Anophelines generally feed just once per gonotrophic (oviposition cycle. Newly emerged insects, however, may feed two or more times during their first oviposition cycle thus increasing the likelihood of becoming infected. The reasons for multiple feeding are not clearly understood and it is still uncertain whether the behaviour is facultative or obligatory. We, therefore, determined survival and sporozoite rates, and examined the behaviour of An. gambiae from the two islands during their first gonotrophic cycle. Methods The wing size of 1,410, abdominal condition of 687, gonotrophic age and mated status of 7,264 female M form An. gambiae collected by light-trap, landing catch, resting outdoors or in copula, was determined from four sites in the archipelago. Sporozoite rates assessed by ELISA in 15,533 females from São Tomé and 2,111 from Príncipe were determined. Results Estimated survival rates ranged between 0.834–0.849 per day in São Tomé and 0.801–0.818 per day in Príncipe. Sporozoite rates of 0.63% in São Tomé were significantly higher than the 0.24% from Príncipe. Overall 49% of females mated on the second night after emergence before feeding, and 51% on the third night and thus fed before mating. The likelihood of mating before feeding increased with wing size. None of the 3,776 parous insects collected showed evidence of recent mating. All but two of the 198 females collected in copula had undeveloped ovaries. Mean wing sizes and the number of insects collected in a sentinel light-trap varied but the proportion of newly emerged insects in the collection did not. The estimated survival rate of the smallest

Limnological and botanical characterization of larval habitats for two primary malarial vectors, Anopheles albimanus and Anopheles pseudopunctipennis, in coastal areas of Chiapas State, Mexico.

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Savage, H M; Rejmankova, E; Arredondo-Jim'enez, J I; Roberts, D R; Rodr'iguez, M H

1990-12-01

Field surveys of mosquito breeding sites on the Pacific coastal plain and foothill regions of southern Chiapas, Mexico, were carried out in the dry and wet seasons of 1988. At each site, selected environmental variables were measured or estimated, presence and percent cover of aquatic plants recorded, a water sample collected for subsequent analyses, and 10-30 dips made for mosquito larvae. Logistic regression and discriminant analyses revealed that the occurrence of Anopheles albimanus larvae in both the wet and dry seasons was positively associated with planktonic algae and negatively associated with altitude. In the dry season, An. albimanus larvae were largely restricted to the margins of permanent water bodies and were associated with the presence of floating plants, particularly Eichhornia crassipes. During the wet season An. albimanus larvae were positively associated with emergent plants, particularly seasonally flooded Cyperaceae, and phosphorus (PO4) concentrations, and were negatively associated with abundant filamentous algae, high levels of total suspended solids (TSS) and Salvinia. In the dry season, An. pseudopunctipennis larvae were positively associated with filamentous algae, altitude and the presence of Heteranthera if encountered in a riverine setting, and were negatively associated with water depth. During the wet season, flooding eliminated typical flood plain An. pseudopunctipennis habitats, and larvae were rarely encountered.

Efek Beauveria bassiana pada Anopheles maculatus Fase Aquatik di Laboratorium

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Bina Ikawati

2017-07-01

Full Text Available AbstractBeauveria bassiana can be used both as for controlling agricultural insect and protecting health. Thisstudy aims to examine the effects of aquatic phase of B. bassiana on An. maculatus in the laboratory.Samples were eggs and larvae of An. maculatus reared from Banjarnegara colony. Eggs and instarlarvae II, III and IV and their control which consisted of 10 larvae/eggs were replicated six times andcontacted to B. bassiana spores for 15 minutes and then transferred to aquades to be maintained forobservation, in every 24 hours as long as 192 hours (8 days. Probit analysis found that applicationof B. bassiana caused damage of external coat of eggs and inhibited >60% of unhatched eggs. Lethaldosage was dosage spores of 1,713x107 (16 days, whereas the lethal dose required to make 50% ofunperached eggs is a dose of spore concentration of 1,361x107 (11.6 days. Higher concentrationswill be needed to know the faster effects B bassiana on An. maculatus larvae or eggs.Key words: Beauveria bassiana, Anopheles maculatus, aquatic phase, laboratory AbstrakBeauveria bassiana dapat digunakan baik sebagai pengendali serangga pertanian maupun kesehatan.Penelitian ini bertujuan mengkaji efek B. bassiana terhadap An. maculatus pada fase akuatik dilaboratorium. Sampel uji berupa telur dan larva An. maculatus dari koloni Banjarnegara. Telur danlarva instar II,III dan IV serta kontrol masing-masing sebanyak 10 ekor/butir dengan replikasi 6 kalidikontakkan dengan spora B. bassiana selama 15 menit dan selanjutnya dipindahkan ke aquadesuntuk dipelihara untuk dilakukan pengamatan, pengamatan dengan mikroskop compound terutamapada larva yang lemah/mati setiap 24 jam selama 192 jam (8 hari. Analisis probit membuktikanbahwa aplikasi B. bassiana pada telur menimbulkan efek kerusakan pada lapisan luar telur diketahuidari pengamatan dengan mikroskop serta mampu menghambat >60% telur tidak menetas. Simulasidengan análisis probit menunjukkan lethal dose yang dibutuhkan

The genetics and ecology of male reproductive investments in the malaria mosquito Anopheles gambiae s.s

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Ekechukwu, Nkuru Esther

2015-01-01

Malaria continues to be a major global health problem due to high mortality and morbidity rate in endemic regions. An. gambiae s.s is the major vector in endemic African countries. About 198 million cases of malaria were recorded globally in 2013 and this have led to over 584 000 deaths. Different measures have been implemented in order to reduce and control the transmission rate. However, the drug resistant parasites and insecticide resistant mosquitoes have created problems towards achievin...

Spatial clustering and longitudinal variation of Anopheles darlingi (Diptera: Culicidae) larvae in a river of the Amazon: the importance of the forest fringe and of obstructions to flow in frontier malaria.

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Barros, F S M; Arruda, M E; Gurgel, H C; Honório, N A

2011-12-01

Deforestation has been linked to a rise in malaria prevalence. In this paper, we studied longitudinally 20 spots, including forested and deforested portions of a temporary river in a malarigenous frontier zone. Larval habitat parameters influencing distribution of Anopheles darlingi (Diptera: Culicidae) larvae were studied. We observed that larvae were clustered in forested-deforested transitions. For the first time in the literature, it was verified that parameters determining larval distribution varied from deforested to forested areas. The proximity to human dwellings was also a significant factor determining distribution, but larvae was most importantly associated with a previously undescribed parameter, the presence of small obstructions to river flow, such as tree trunks within the river channel, which caused pooling of water during the dry season ('microdams'). In deforested areas, the most important factor determining distribution of larvae was shade (reduced luminance). Larvae were absent in the entire studied area during the wet season and present in most sites during the dry season. During the wet-dry transition, larvae were found sooner in areas with microdams, than in other areas, suggesting that flow obstruction prolongs the breeding season of An. darlingi. Adult mosquito densities and malaria incidence were higher during the dry season. Our data correlate well with the published literature, including the distribution of malaria cases near the forest fringes, and has permitted the creation of a model of An. darlingi breeding, where preference for sites with reduced luminance, human presence and microdams would interact to determine larval distribution.

The Romanomermis iyengari parasite for Anopheles pseudopunctipennis suppression in natural habitats in Oaxaca State, Mexico

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Santamarina Mijares Alberto

1999-01-01

Full Text Available In September and November 1996 Romanomermis iyengari Welch, a parasite of larval mosquitoes, was released in 44 natural larval habitat sites of Anopheles pseudopunctipennis Theobald in an attempt to reduce the larval populations of this important malaria vector. The selected treatment sites ranged in size from 5 to 500 m². The study was carried out in Pochutla District of Oaxaca State, on the Pacific coast of Mexico. Chemical pesticides to reduce vector populations have been the principal tool in malaria suppression campaigns. However, the excessive use of these chemicals has created pesticide resistance and other serious collateral problems. Therefore, a biological control project using agents that are pathogens of Anopheles larvae was initiated in 1996. The principal objective was to establish mass rearing capacities for R. iyengari. Detailed methodology for rearing and introducing these nematodes into mosquito larval habitats was established at the National Polytechnic Institute of Oaxaca State. Before application of the parasites to larval habitats, site characteristics were determined, including size, depth, aquatic vegetation, salinity, pH, conductivity, temperature, and pretreatment larval density. With a compressed air sprayer, infective mermithid parasites were released at rates of either 2000 or 3000/m², and the parasites produced high levels of infection. Anopheles populations were sampled 72 h posttreatment, and the larvae obtained were taken to the laboratory and examined through microscopic dissection to determine infection levels and mean parasitism. Nematode parasitism ranged from 85 to 100% at all the treatment sites, even though no previous information concerning field parasitism of An. pseudopunctipennis by R. iyengari has been reported. In addition, a significant reduction of mosquito larval density at the treatment sites was found five days after the nematode application. Levels of parasitism were indicative of the number

Thermal limits of wild and laboratory strains of two African malaria vector species, Anopheles arabiensis and Anopheles funestus

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Lyons Candice L

2012-07-01

Full Text Available Abstract Background Malaria affects large parts of the developing world and is responsible for almost 800,000 deaths annually. As climates change, concerns have arisen as to how this vector-borne disease will be impacted by changing rainfall patterns and warming temperatures. Despite the importance and controversy surrounding the impact of climate change on the potential spread of this disease, little information exists on the tolerances of several of the vector species themselves. Methods Using a ramping protocol (to assess critical thermal limits - CT and plunge protocol (to assess lethal temperature limits - LT information on the thermal tolerance of two of Africa’s important malaria vectors, Anopheles arabiensis and Anopheles funestus was collected. The effects of age, thermal acclimation treatment, sex and strain (laboratory versus wild adults were investigated for CT determinations for each species. The effects of age and sex for adults and life stage (larvae, pupae, adults were investigated for LT determinations. Results In both species, females are more tolerant to low and high temperatures than males; larvae and pupae have higher upper lethal limits than do adults. Thermal acclimation of adults has large effects in some instances but small effects in others. Younger adults tend to be more tolerant of low or high temperatures than older age groups. Long-standing laboratory colonies are sufficiently similar in thermal tolerance to field-collected animals to provide reasonable surrogates when making inferences about wild population responses. Differences between these two vectors in their thermal tolerances, especially in larvae and pupae, are plausibly a consequence of different habitat utilization. Conclusions Limited plasticity is characteristic of the adults of these vector species relative to others examined to date, suggesting limited scope for within-generation change in thermal tolerance. These findings and the greater tolerance

Hybrid sterility in crosses between two Brazilian sibling species of the Anopheles albitarsis complex.

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Fontoura, Nathalia Giglio; Araki, Alejandra Saori; Van Der Maas Azevedo, Renata; Galardo, Allan Kardec Ribeiro; Peixoto, Alexandre Afranio; Lima, José Bento Pereira

2014-12-04

Complexes of cryptic species are common in several taxa and this is also the case in the Anopheles genus, a group including all known human malaria vectors. The Anopheles albitarsis complex comprises at least nine cryptic species, some of which are implicated as vectors of human malaria. Several different types of data have been generated for this species complex such as cytogenetics, alloenzymes, morphological and feeding behavioral, hybridization experiments, RAPD-PCR and RFLP and mitochondrial and nuclear markers. Studies focused on its postzygotic isolation are still somewhat rare in the literature despite their importance to understand the speciation process and the level of gene flow potentially occurring among the different sibling species. Hybridization experiments between Anopheles albitarsis s.s. and Anopheles marajoara, as well as backcrosses between hybrids and Anopheles albitarsis s.s., were performed using the induced mating technique. Results were compared to intraspecific crosses. Larva-to-adult viability and sex ratio were also assessed. Male hybrids show very low insemination rates and nearly complete sterility, apparently due to abnormalities in their reproductive organs. Evidence of partial sterility among the hybrid females was also observed. Our data indicated that Anopheles albitarsis s.s. and Anopheles marajoara show a high level of postzygotic isolation with a strong hybrid male sterility. This result is consistent with the Haldane's rule which states that in interspecific crosses the heterogametic sex is the first to be affected. However, the fact that the females are not completely sterile raises the possibility of introgression between these two siblings species.

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

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Lwetoijera Dickson W

2009-12-01

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

Attractiveness of MM-X Traps Baited with Human or Synthetic Odor to Mosquitoes (Diptera: Culicidae) in The Gambia

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QIU, YU TONG; SMALLEGANGE, RENATE C.; TER BRAAK, CAJO J. F.; SPITZEN, JEROEN; VAN LOON, JOOP J. A.; JAWARA, MUSA; MILLIGAN, PAUL; GALIMARD, AGNES M.; VAN BEEK, TERIS A.; KNOLS, BART G. J.; TAKKEN, WILLEM

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

Agriculture and the promotion of insect pests: rice cultivation in river floodplains and malaria vectors in The Gambia

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Louca Vasilis

2009-07-01

Full Text Available Abstract Background Anthropogenic modification of natural habitats can create conditions in which pest species associated with humans can thrive. In order to mitigate for these changes, it is necessary to determine which aspects of human management are associated with the promotion of those pests. Anopheles gambiae, the main Africa malaria vector, often breeds in rice fields. Here the impact of the ancient practice of 'swamp rice' cultivation, on the floodplains of the Gambia River, on the production of anopheline mosquitoes was investigated. Methods Routine surveys were carried out along 500 m transects crossing rice fields from the landward edge of the floodplains to the river during the 2006 rainy season. Aquatic invertebrates were sampled using area samplers and emergence traps and fish sampled using nets. Semi-field experiments were used to investigate whether nutrients used for swamp rice cultivation affected mosquito larval abundance. Results At the beginning of the rainy season rice is grown on the landward edge of the floodplain; the first area to flood with fresh water and one rich in cattle dung. Later, rice plants are transplanted close to the river, the last area to dry out on the floodplain. Nearly all larval and adult stages of malaria vectors were collected 0–100 m from the landward edge of the floodplains, where immature rice plants were grown. These paddies contained stagnant freshwater with high quantities of cattle faeces. Semi-field studies demonstrated that cattle faeces nearly doubled the number of anopheline larvae compared with untreated water. Conclusion Swamp rice cultivation creates ideal breeding sites for malaria vectors. However, only those close to the landward edge harboured vectors. These sites were productive since they were large areas of standing freshwater, rich in nutrients, protected from fish, and situated close to human habitation, where egg-laying mosquitoes from the villages had short distances to

Light manipulation of mosquito behaviour: acute and sustained photic suppression of biting activity in the Anopheles gambiae malaria mosquito.

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Sheppard, Aaron D; Rund, Samuel S C; George, Gary F; Clark, Erin; Acri, Dominic J; Duffield, Giles E

2017-06-16

Host-seeking behaviours in anopheline mosquitoes are time-of-day specific, with a greater propensity for nocturnal biting. We investigated how a short exposure to light presented during the night or late day can inhibit biting activity and modulate flight activity behaviour. Anopheles gambiae (s.s.), maintained on a 12:12 LD cycle, were exposed transiently to white light for 10-min at the onset of night and the proportion taking a blood meal in a human biting assay was recorded every 2 h over an 8-h duration. The pulse significantly reduced biting propensity in mosquitoes 2 h following administration, in some trials for 4 h, and with no differences detected after 6 h. Conversely, biting levels were significantly elevated when mosquitoes were exposed to a dark treatment during the late day, suggesting that light suppresses biting behaviour even during the late daytime. These data reveal a potent effect of a discrete light pulse on biting behaviour that is both immediate and sustained. We expanded this approach to develop a method to reduce biting propensity throughout the night by exposing mosquitoes to a series of 6- or 10-min pulses presented every 2 h. We reveal both an immediate suppressive effect of light during the exposure period and 2 h after the pulse. This response was found to be effective during most times of the night: however, differential responses that were time-of-day specific suggest an underlying circadian property of the mosquito physiology that results in an altered treatment efficacy. Finally, we examined the immediate and sustained effects of light on mosquito flight activity behaviour following exposure to a 30-min pulse, and observed activity suppression during early night, and elevated activity during the late night. As mosquitoes and malaria parasites are becoming increasingly resistant to insecticide and drug treatment respectively, there is a necessity for the development of innovative control strategies beyond insecticide

Estacionalidad de la densidad larval del mosquito Anopheles aquasalis (Diptera: Culicidae y otros insectos asociados a su hábitat en Sucre, Venezuela

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Jesús Berti

2010-06-01

Full Text Available Anopheles aquasalis es considerado como el principal vector de malaria humana en el norte de Venezuela. El estudio longitudinal se llevó a cabo en áreas costeras de la Península de Paria, estado Sucre. El hábitat de las larvas de A. aquasalis fue clasificado como: manglar salobre y pantano herbáceo. Muestreos para recolectar larvas de mosquitos e insectos asociados se realizaron mensualmente en ambos criaderos desde enero hasta diciembre de 1999 (30 muestras. Simultáneamente se midieron cinco variables seleccionadas del agua: conductividad, salinidad, oxígeno disuelto, temperatura y pH. En ambos criaderos de determinaron las variaciones estacionales y temporales de las larvas de A. aquasalis e insectos acuáticos. Para el período de estudio, la abundancia de larvas fue mayor en el manglar. El análisis de correspondencia mostró una fuerte relación entre algunos factores químicos del agua y la abundancia de las larvas. La abundancia de las larvas de A. aquasalis en ambas épocas, se correlacionó positivamente con la salinidad del agua, pH y conductividad, y negativamente con el oxígeno disuelto, en la estación seca. La presencia de larvas se correlacionó positivamente con la presencia de Avicennia germinans. En el manglar existió una asociación positiva entre la abundancia de larvas y la abundancia de la familia Scirtidae y una correlación negativa entre la abundancia de larvas y la precipitación mensual (Spearman, así como una correlación negativa significativa entre la abundancia de Gerridae y la precipitación mensual. En el pantano herbáceo, no había asociaciones significativas entre la abundancia de las larvas de A. aquasalis y la abundancia de otros insectos acuáticos asociados al hábitat.Larval seasonality of the mosquito Anopheles aquasalis (Diptera: Culicidae and other insects associated to its habitat in Sucre, Venezuela. Anopheles aquasalis Curry is considered the main vector of human malaria in Northern

FAUNA ANOPHELES DI DAERAH PANTAI BEKAS HUTAN MANGROVE KECAMATAN PADANG CERMIN KABUPATEN LAMPUNG SELATAN

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N. Sushanti Idris-Idram

2012-09-01

Full Text Available Intensive mosquito collections were carried out in two villages in subdistrict of Padangcermin during 1992-1993. The method of mosquito collections consisted of night landing on man indoor and outdoor, night resting indoor and outdoor around cattle shelters, light trap in cattle shelters, daytime resting indoor and outdoor, as well as larvae collections to identify anophelines breeding sites. Sixteen anophelines i.e. Anopheles sundaicus, An. subpictus, An. vagus, An. indefinitus, An. nigerrimus, An. peditaeniatus, An. kochi, An. barbirostris, An. bambumbrosus, An. annularis, An. separatus, An. tesselatus, An. aconitus, An. umbrosus, An. leucosphyrus and An. letifer were collected. Among these mosquitos, An. sundaicus was found predominant, followed by An. vagus and An. subpictus. Other species were collected in small numbers. The behavior of Anopheles sundaicus, An. subpictus and An. vagus were exophagic and endophilic. The larvae of An. sundaicus was found only in brackish standing water such as abandoned shrimp ponds, An. subpictus in brackish standing water as well as fresh standing water, while An. vagus was found only in fresh standing water. Breeding sites of An. sundaicus was characterized by pond with floating algae while An. subpictus and An. vagus were not depending on vegetation.

Plasmodium falciparum infection rates for some Anopheles spp. from Guinea-Bissau, West Africa [v2; ref status: indexed, http://f1000r.es/4n3

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Michelle R. Sanford

2014-11-01

Full Text Available Presence of Plasmodium falciparum circumsporozoite protein (CSP was detected by enzyme linked immunosorbent assay (ELISA in a sample of Anopheles gambiae s.s., A. melas and A. pharoensis collected in Guinea-Bissau during October and November 2009. The percentage of P. falciparum infected samples (10.2% overall; 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.

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

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Gilioli Gianni

2011-10-01

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

Mosquitoes rely on their gut microbiota for development

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Coon, Kerri L.; Vogel, Kevin J.; Brown, Mark R.; Strand, Michael R.

2014-01-01

Field studies indicate adult mosquitoes (Culicidae) host low diversity communities of bacteria that vary greatly among individuals and species. In contrast, it remains unclear how adult mosquitoes acquire their microbiome, what influences community structure, and whether the microbiome is important for survival. Here we used pyrosequencing of 16S rRNA to characterize the bacterial communities of three mosquito species reared under identical conditions. Two of these species, Aedes aegypti and Anopheles gambiae, are anautogenous and must blood feed to produce eggs, while one, Georgecraigius atropalpus, is autogenous and produces eggs without blood feeding. Each mosquito species contained a low diversity community comprised primarily of aerobic bacteria acquired primarily from the aquatic habitat in which larvae developed. Our results suggested the communities in Ae. aegypti and An. gambiae larvae share more similarities with one another than with Ge. atropalpus. Studies with Ae. aegypti also strongly suggested that adults transstadially acquired several members of the larval bacterial community, but only four genera of bacteria present in blood fed females were detected on eggs. Functional assays showed that axenic larvae of each species failed to develop beyond the first instar. Experiments with Ae. aegypti indicated several members of the microbial community and Escherichia coli successfully colonized axenic larvae and rescued development. Overall, our results provide new insights about the acquisition and structure of bacterial communities in mosquitoes. They also indicate three mosquito species spanning the breadth of the Culicidae depend on their gut microbiome for development. PMID:24766707

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

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Koffi Alphonsine A

2011-06-01

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

Differential attractiveness of humans to the African malaria vector Anopheles gambiae Giles : effects of host characteristics and parasite infection

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Mukabana, W.R.

2002-01-01

The results of a series of studies designed to understand the principal factors that determine the differential attractiveness of humans to the malaria vector Anopheles