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1.
Parasit Vectors ; 13(1): 202, 2020 Apr 19.
Artigo em Inglês | MEDLINE | ID: mdl-32307003

RESUMO

BACKGROUND: Malaria vector mosquitoes acquire midgut microbiota primarily from their habitat. The homeostasis of these microbial communities plays an essential role in the mosquito longevity, the most essential factor in the mosquito vectorial capacity. Our recent study revealed that silencing genes involved in regulation of the midgut homeostasis including FN3D1, FN3D3 and GPRGr9 reduced the survival of female adult Anopheles arabiensis mosquitoes. In the present study, we investigate the stability of the gene silencing efficiency of mosquitoes reared in three different breeding conditions representing distinct larval habitat types: town brick pits in Jimma, flood pools in the rural land of Asendabo and roadside pools in Wolkite. METHODS: First-instar larvae of An. arabiensis mosquitoes were reared separately using water collected from the three breeding sites. The resulting adult females were micro-injected with dsRNA targeting the FN3D1 gene (AARA003032) and their survival was monitored. Control mosquitoes were injected with dsRNA Lacz. In addition, the load of midgut microbiota of these mosquitoes was determined using flow cytometry. RESULTS: Survival of naïve adult female mosquitoes differed between the three sites. Mosquitoes reared using water collected from brick pits and flood pools survived longer than mosquitoes reared using water collected from roadside. However, the FN3D1 gene silencing effect on survival did not differ between the three sites. CONCLUSIONS: The present study revealed that the efficacy of FN3D1 gene silencing is not affected by variation in the larval habitat. Thus, silencing this gene has potential for application throughout sub-Saharan Africa.


Assuntos
Anopheles/genética , Domínio de Fibronectina Tipo III/genética , Interferência de RNA/fisiologia , Animais , Anopheles/fisiologia , Cruzamento , Ecossistema , Larva/genética , Larva/fisiologia , Controle de Mosquitos/métodos , Mosquitos Vetores/genética , Mosquitos Vetores/fisiologia
2.
Lancet ; 395(10232): 1292-1303, 2020 04 18.
Artigo em Inglês | MEDLINE | ID: mdl-32305094

RESUMO

BACKGROUND: Long-lasting insecticidal nets (LLINs) are the primary malaria prevention tool, but their effectiveness is threatened by pyrethroid resistance. We embedded a pragmatic cluster-randomised trial into Uganda's national LLIN campaign to compare conventional LLINs with those containing piperonyl butoxide (PBO), a synergist that can partially restore pyrethroid susceptibility in mosquito vectors. METHODS: 104 health sub-districts, from 48 districts in Uganda, were randomly assigned to LLINs with PBO (PermaNet 3.0 and Olyset Plus) and conventional LLINs (PermaNet 2.0 and Olyset Net) by proportionate randomisation using an iterative process. At baseline 6, 12, and 18 months after LLIN distribution, cross-sectional surveys were done in 50 randomly selected households per cluster (5200 per survey); a subset of ten households per cluster (1040 per survey) were randomly selected for entomological surveys. The primary outcome was parasite prevalence by microscopy in children aged 2-10 years, assessed in the as-treated population at 6, 12, and 18 months. This trial is registered with ISRCTN, ISRCTN17516395. FINDINGS: LLINs were delivered to households from March 25, 2017, to March 18, 2018, 32 clusters were randomly assigned to PermaNet 3.0, 20 to Olyset Plus, 37 to PermaNet 2.0, and 15 to Olyset Net. In the as-treated analysis, three clusters were excluded because no dominant LLIN was received, and four clusters were reassigned, resulting in 49 PBO LLIN clusters (31 received PermaNet 3.0 and 18 received Olyset Plus) and 52 non-PBO LLIN clusters (39 received PermaNet 2.0 and 13 received Olyset Net). At 6 months, parasite prevalence was 11% (386/3614) in the PBO group compared with 15% (556/3844) in the non-PBO group (prevalence ratio [PR] adjusted for baseline values 0·74, 95% CI 0·62-0·87; p=0·0003). Parasite prevalence was similar at month 12 (11% vs 13%; PR 0·73, 95% CI 0·63-0·85; p=0·0001) and month 18 (12% vs 14%; PR 0·84, 95% CI 0·72-0·98; p=0·029). INTERPRETATION: In Uganda, where pyrethroid resistance is high, PBO LLINs reduced parasite prevalence more effectively than did conventional LLINs for up to 18 months. This study provides evidence needed to support WHO's final recommendation on use of PBO LLINs. FUNDING: The Against Malaria Foundation, UK Department for International Development, Innovative Vector Control Consortium, and Bill and Melinda Gates Foundation.


Assuntos
Mosquiteiros Tratados com Inseticida/estatística & dados numéricos , Malária/prevenção & controle , Sinergistas de Praguicidas/farmacologia , Butóxido de Piperonila/farmacologia , Animais , Anopheles/parasitologia , Anopheles/fisiologia , Criança , Pré-Escolar , Estudos Transversais , Feminino , Humanos , Resistência a Inseticidas , Malária/sangue , Masculino , Mosquitos Vetores/parasitologia , Mosquitos Vetores/fisiologia , Uganda
3.
Parasit Vectors ; 13(1): 55, 2020 Feb 10.
Artigo em Inglês | MEDLINE | ID: mdl-32041663

RESUMO

BACKGROUND: Nyssorhynchus darlingi (also known as Anopheles darlingi) is the primary malaria vector in the Amazon River Basin. In Brazil, analysis of single nucleotide polymorphisms (SNPs) previously detected three major population clusters, and a common garden experiment in a laboratory setting revealed significant population variation in life history traits. Increasing temperatures and local level variation can affect life history traits, i.e. adult longevity, that alter vectorial capacity with implications for malaria transmission in Ny. darlingi. METHODS: We investigated the population structure of Ny. darlingi from 7 localities across Brazil utilizing SNPs and compared them to a comprehensive Ny. darlingi catalog. To test the effects of local level variation on life history traits, we reared F1 progeny from the 7 localities at three constant temperatures (20, 24 and 28 °C), measuring key life history traits (larval development, food-starved adult lifespan, adult size and daily survival). RESULTS: Using nextRAD genotyping-by-sequencing, 93 of the field-collected Ny. darlingi were genotyped at 33,759 loci. Results revealed three populations (K = 3), congruent with major biomes (Amazonia, Cerrado and Mata Atlântica), with greater FST values between biomes than within. In the life history experiments, increasing temperature reduced larval development time, adult lifespan, and wing length in all localities. The variation of family responses for all traits within four localities of the Amazonia biome was significant (ANOVA, P < 0.05). Individual families within localities revealed a range of responses as temperature increased, for larval development, adult lifespan, wing length and survival time. CONCLUSIONS: SNP analysis of several Brazilian localities provided results in support of a previous study wherein populations of Ny. darlingi were clustered by three major Brazilian biomes. Our laboratory results of temperature effects demonstrated that population variation in life history traits of Ny. darlingi exists at the local level, supporting previous research demonstrating the high plasticity of this species. Understanding this plasticity and inherent variation between families of Ny. darlingi at the local level should be considered when deploying intervention strategies and may improve the likelihood of successful malaria elimination in South America.


Assuntos
Anopheles/fisiologia , Traços de História de Vida , Fenótipo , Temperatura , Adaptação Fisiológica/fisiologia , Animais , Brasil , Ecossistema , Malária/transmissão , Mosquitos Vetores/fisiologia , Dinâmica Populacional
4.
Science ; 367(6478): 681-684, 2020 02 07.
Artigo em Inglês | MEDLINE | ID: mdl-32029627

RESUMO

Mosquitoes transmit pathogens that kill >700,000 people annually. These insects use body heat to locate and feed on warm-blooded hosts, but the molecular basis of such behavior is unknown. Here, we identify ionotropic receptor IR21a, a receptor conserved throughout insects, as a key mediator of heat seeking in the malaria vector Anopheles gambiae Although Ir21a mediates heat avoidance in Drosophila, we find it drives heat seeking and heat-stimulated blood feeding in Anopheles At a cellular level, Ir21a is essential for the detection of cooling, suggesting that during evolution mosquito heat seeking relied on cooling-mediated repulsion. Our data indicate that the evolution of blood feeding in Anopheles involves repurposing an ancestral thermoreceptor from non-blood-feeding Diptera.


Assuntos
Anopheles/fisiologia , Temperatura Corporal , Evolução Molecular , Comportamento de Busca por Hospedeiro/fisiologia , Temperatura Alta , Receptores Ionotrópicos de Glutamato/fisiologia , Termorreceptores/fisiologia , Animais , Anopheles/genética , Sangue , Feminino , Camundongos , Mutação , Receptores Ionotrópicos de Glutamato/genética
5.
PLoS One ; 15(2): e0228576, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049962

RESUMO

Anopheles arabiensis is an opportunistic malaria vector that rests and feeds outdoors, circumventing current vector control methods. Furthermore, this vector will readily feed on animal as well as human hosts. Targeting the vector, while feeding on animals, can provide an additional intervention for the current vector control activities. Agricultural animals are regularly vaccinated with recombinant proteins for the control of multiple endo- and ecto-parasitic infestations. The use of a Subolesin-vaccine showed a mark reduction in tick reproductive fitness. The orthologous gene of Subolesin, called Akirin in insects, might provide a valuable species-specific intervention against outdoor biting An. arabiensis. However, the biological function of this nuclear protein has not yet been investigated in this mosquito. The effects on An. arabiensis lifetable parameters were evaluated after Akirin was knocked down using commercial small-interfering RNA (siRNA) and in vitro transcribed double-stranded RNA (dsRNA). The siRNA mediated interference of Akirin significantly reduced fecundity by 17%, fertility by 23% and longevity by 32% when compared to the controls in the female mosquitoes tested. Similarly, dsRNA treatment had a 25% decrease in fecundity, 29% decrease in fertility, and 48% decrease in longevity, when compared to the control treatments. Mosquitoes treated with Akirin dsRNA had a mean survival time of 15-days post-inoculation, which would impact on their ability to transmit malaria parasites. These results strongly suggest that Akirin has a pleiotropic function in An. arabiensis longevity and reproductive fitness.


Assuntos
Anopheles/genética , Fertilidade/genética , Proteínas de Insetos/genética , Longevidade/genética , Animais , Anopheles/fisiologia , Feminino , Masculino , Interferência de RNA
6.
Parasit Vectors ; 13(1): 4, 2020 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-31910892

RESUMO

BACKGROUND: Members of the Anopheles gambiae complex breed in clean, sunlit temporary bodies of water. Anthropogenic pollution is, however, altering the breeding sites of the vectors with numerous biological effects. Although the effects of larval metal pollution have previously been examined, this study aims to assess the transgenerational effects of larval metal pollution on the major malaria vector An. arabiensis. METHODS: Two laboratory strains of An. arabiensis, SENN (insecticide-susceptible) and SENN-DDT (insecticide-resistant), were used in this study. After being bred in water polluted with either cadmium chloride, copper nitrate or lead nitrate, several life history characteristics that can have epidemiological implications (fertility, apoptotic damage to reproductive structures, adult longevity and insecticide tolerance) were examined in the adults and compared to those of adults bred in clean water. RESULTS: All metal treatments reduced fecundity in SENN, but only lead treatment reduced fertility in SENN-DDT. Cadmium chloride exposure resulted in apoptosis and deformation of the testes in both strains. After breeding generation F0 in polluted water, F1 larvae bred in clean water showed an increase in longevity in SENN-DDT adult females. In contrast, after breeding the F0 generation in polluted water, longevity was reduced after cadmium and copper exposure in the F1 generation. Larval metal exposure resulted in an increase in insecticide tolerance in adults of the SENN strain, with SENN-DDT adults gaining the greatest fold increase in insecticide tolerance. CONCLUSIONS: This study demonstrates that a single exposure to metal pollution can have transgenerational effects that are not negated by subsequent breeding in clean water.


Assuntos
Anopheles/crescimento & desenvolvimento , Inseticidas/farmacologia , Larva/efeitos dos fármacos , Metais/farmacologia , Poluentes da Água/farmacologia , Animais , Anopheles/efeitos dos fármacos , Anopheles/fisiologia , Resistência a Medicamentos , Feminino , Fertilidade/efeitos dos fármacos , Larva/crescimento & desenvolvimento , Masculino , Reprodução/efeitos dos fármacos
7.
Infect Dis Poverty ; 9(1): 9, 2020 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-31987056

RESUMO

BACKGROUND: Irrigated agriculture is key to increase agricultural productivity and ensure food security in Africa. However, unintended negative public health impacts (e.g. malaria) of such environmental modification have been a challenge. This study assessed the diversity and distribution of breeding habitats of malaria vector mosquitoes around Arjo-Dedessa irrigation development site in Southwest Ethiopia. METHODS: Anopheline mosquito larvae were surveyed from two agroecosystems, 'irrigated' and 'non-irrigated' areas during the dry (December 2017-February 2018) and wet (June 2018-August 2018) seasons. Mosquito habitat diversity and larval abundance were compared between the irrigated and non-irrigated areas. The association between anopheline mosquito larvae occurrence and environmental parameters was analysed using Pearson chi-square. Multiple logistic regression analysis was used to determine primary parameters that influence the occurrence of anopheline larvae. RESULTS: Overall, 319 aquatic habitats were surveyed during the study period. Around 60% (n = 152) of the habitats were positive for anopheline mosquito larvae, of which 63.8% (n = 97) and 36.2% (n = 55) were from irrigated and non-irrigated areas, respectively. The number of anopheline positive habitats was two-fold higher in irrigated than non-irrigated areas. Anopheline larval abundance in the irrigated area was 16.6% higher than the non-irrigated area. Pearson's chi-square analysis showed that season (χ2 = 63.122, df = 1, P < 0.001), agroecosystem (being irrigated or non-irrigated) (χ2 = 6.448, df = 1, P = 0.011), and turbidity (χ2 = 7.296, df = 2, P = 0.025) had a significant association with larval anopheline occurrence. CONCLUSIONS: The study showed a higher anopheline mosquito breeding habitat diversity, larval occurrence and abundance in the irrigated than non-irrigated areas in both dry and wet seasons. This indicates that irrigation development activities contribute to proliferation of suitable mosquito breeding habitats that could increase the risk of malaria transmission. Incorporating larval source management into routine malaria vector control strategies could help reduce mosquito population density and malaria transmission around irrigation schemes.


Assuntos
Irrigação Agrícola , Distribuição Animal , Anopheles/fisiologia , Ecossistema , Controle de Mosquitos , Mosquitos Vetores/fisiologia , Animais , Anopheles/crescimento & desenvolvimento , Etiópia , Larva/fisiologia , Mosquitos Vetores/crescimento & desenvolvimento , Estações do Ano
8.
Nat Commun ; 10(1): 5634, 2019 12 10.
Artigo em Inglês | MEDLINE | ID: mdl-31822677

RESUMO

The blood-feeding behavior of Anopheles females delivers essential nutrients for egg development and drives parasite transmission between humans. Plasmodium growth is adapted to the vector reproductive cycle, but how changes in the reproductive cycle impact parasite development remains unclear. Here, we show that the bloodmeal-induced miR-276-5p fine-tunes the expression of branched-chain amino acid transferase to terminate the reproductive cycle. Silencing of miR-276 prolongs high rates of amino acid (AA) catabolism and increases female fertility, suggesting that timely termination of AA catabolism restricts mosquito investment into reproduction. Prolongation of AA catabolism in P. falciparum-infected females also compromises the development of the transmissible sporozoite forms. Our results suggest that Plasmodium sporogony exploits the surplus mosquito resources available after reproductive investment and demonstrate the crucial role of the mosquito AA metabolism in within-vector parasite proliferation and malaria transmission.


Assuntos
Anopheles/fisiologia , MicroRNAs/metabolismo , Plasmodium falciparum/crescimento & desenvolvimento , Aminoácidos/metabolismo , Animais , Anopheles/efeitos dos fármacos , Sequência de Bases , Ecdisona/farmacologia , Corpo Adiposo/metabolismo , Feminino , Inativação Gênica , MicroRNAs/genética , Modelos Biológicos , Reprodução/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Esteroides/metabolismo , Transaminases/metabolismo
9.
Malar J ; 18(1): 396, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31796010

RESUMO

India has committed to eliminate malaria by 2030. The national framework for malaria elimination released by the Government of India plans to achieve this goal through strategic planning in a phased manner. Since vector control is a major component of disease management and vector elimination, it requires a thorough understanding of the biology and bionomics of malaria vectors exhibiting definite distribution patterns in diverse ecosystems in the country. Although a wealth of information is available on these aspects, lesser-known data are on biting time and rhythm, and the magnitude of outdoor transmission by the vectors which are crucial for effective implementation of the key vector control interventions. Most of the data available for the vector species are at sensu lato level, while the major vectors are species complexes and their members distinctly differ in biological characters. Furthermore, the persistent use of insecticides in indoor residual spray and long-lasting insecticidal nets has resulted in widespread resistance in vectors and changes in their behaviour. In this document, challenges in vector control in the Indian context have been identified and possible solutions to overcome the problem are suggested. Adequate addressing of the issues raised would greatly help make a deep dent in malaria transmission and consequently result in disease elimination within the targeted time frame.


Assuntos
Anopheles/fisiologia , Malária/prevenção & controle , Controle de Mosquitos/métodos , Mosquitos Vetores/fisiologia , Animais , Índia , Traços de História de Vida
10.
Infect Dis Poverty ; 8(1): 100, 2019 Dec 03.
Artigo em Inglês | MEDLINE | ID: mdl-31796068

RESUMO

BACKGROUND: The Sahel region of Chad Republic is a prime candidate for malaria pre-elimination. To facilitate pre-elimination efforts in this region, two populations of Anopheles coluzzii from Central Chad Republic were characterized, their insecticide resistance profile and the possible molecular mechanisms driving the resistance in the field investigated. METHODS: Bloodfed female Anopheles gambiae s.l. resting indoor, were collected at N'djamena and Massakory, Chad in 2018 and characterized for species composition, and infection rate was determined using the TaqMan assay. Susceptibility to various insecticides was assessed using WHO tube bioassays. Cone bioassays were conducted using various long-lasting insecticidal nets (LLINs). Results were analysed using Chi Square test. Knockdown resistance (kdr) and ace-1 markers were investigated by TaqMan genotyping. RESULTS: Anopheles coluzzii was the major vector found in N'djamena (100%) and Massakory (~ 94%). No Plasmodium was found in 147 bloodfed F0 An. coluzzii (82 from N'djamena and 65 from Massakory). High intensity pyrethroid resistance was observed with mortalities of < 2% for permethrin, deltamethrin and etofenprox, and with < 50% and < 60% dead following exposure to 10× diagnostic doses of deltamethrin and permethrin, respectively. For both sites, < 10% mortalities were observed with DDT. Synergist bioassays with piperonylbutoxide significantly recovered pyrethroid susceptibility in Massakory populations, implicating CYP450s (mortality = 13.6% for permethrin, χ2 = 22.8, df = 1, P = 0.0006; mortality = 13.0% for deltamethrin, χ2 = 8.8, df = 1, P < 0.00031). Cone-bioassays established complete loss of efficacy of the pyrethroid-based LLINs; and a 100% recovery of susceptibility following exposure to the roof of PermaNet®3.0, containing piperonylbutoxide. Both populations were susceptible to malathion, but high bendiocarb resistance was observed in Massakory population. The absence of ace-1 mutation points to the role of metabolic resistance in the bendiocarb resistance. Both 1014F and 1014S mutations were found in both populations at around 60% and < 20% respectively. Sequencing of intron-1 of the voltage-gated sodium channel revealed a low genetic diversity suggesting reduced polymorphism. CONCLUSIONS: Multiple resistance in An. coluzzii populations from Chad highlight challenges associated with deployment of LLINs and indoor residual spraying (IRS) in the Sahel of this country. The pyrethroid-synergists LLINs (e.g. PermaNet®3.0) and organophosphate-based IRS maybe the alternatives for malaria control in this region.


Assuntos
Anopheles/fisiologia , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Malária/transmissão , Mosquitos Vetores/fisiologia , Animais , Anopheles/efeitos dos fármacos , Anopheles/genética , Chade , Feminino , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/genética
11.
PLoS One ; 14(12): e0226191, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31869350

RESUMO

BACKGROUND: In view of widespread pyrethroid resistance in malaria vectors in Africa, two long-lasting insecticidal nets (LLINs) incorporated with a synergist, piperonyl butoxide (PBO), DawaPlus 3.0 (deltamethrin + PBO in the roof panel; deltamethrin alone in the side panels) and DawaPlus 4.0 (deltamethrin + PBO in all panels), were evaluated in an experimental hut trial in a rice growing irrigated area in Burkina Faso. Efficacy of nets was tested against free-flying malaria vector, Anopheles gambiae s.l., with high pyrethroid resistance involving L1014F kdr and CYP6P3P450 resistance mechanisms. METHODOLOGY: The efficacy of unwashed and 20-times washed DawaPlus 3.0 (polyethylene roof panel with 120 mg/m2 deltamethrin and 440 mg/m2 PBO; polyester side panels with deltamethrin 100 mg/m2) and DawaPlus 4.0 (same composition as roof of DawaPlus 3.0) was evaluated against DawaPlus 2.0 (80 mg/m2 deltamethrin; positive control). Volunteer sleepers and treatments were rotated in huts using a Latin square design on 63 consecutive nights during August-October 2016. Mortality, human blood-feeding inhibition, deterrence and exit rates of An. gambiae s.l. were monitored. PRINCIPAL FINDINGS: Significantly higher rates of mortality and blood-feeding inhibition were observed with unwashed DawaPlus 4.0 (36%; 47.5%) than unwashed DawaPlus 3.0 (11.8%; 33.3%), DawaPlus 2.0 (4.3%; 6.4%) or untreated net (P < 0.05). Washing reduced personal protective efficacy yet PBO-LLINs were more protective and both met the WHO criteria. CONCLUSIONS: The PBO-containing DawaPlus 4.0 significantly protected against An. gambiae s.l. in the study area. Unwashed DawaPlus 3.0 gave low to moderate protection against the positive control. PBO inhibits oxidase action; hence in areas with active malaria transmission having oxidase mechanisms, PBO nets could confer additional personal protection.


Assuntos
Anopheles/efeitos dos fármacos , Mosquiteiros Tratados com Inseticida , Controle de Mosquitos/métodos , Nitrilos/farmacologia , Butóxido de Piperonila/farmacologia , Piretrinas/farmacologia , Agricultura , Animais , Anopheles/fisiologia , Burkina Faso , Desenho de Equipamento , Voo Animal/efeitos dos fármacos , Gossypium , Humanos , Inseticidas/farmacologia , Malária/prevenção & controle , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/fisiologia , Oryza
12.
PLoS One ; 14(12): e0226815, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31887129

RESUMO

Host seeking in the yellow fever mosquito, Aedes aegypti, and the African malaria mosquito, Anopheles coluzzii, relies on specific and generic host-derived odorants. Previous analyses indicate that the behavioral response of these species depends differentially on the presence of carbon dioxide (CO2) and other constituents in human breath for activation and attraction. In this study, we use a flight tube assay and electrophysiological analysis to assess the role of acetone, a major component of exhaled human breath, in modulating the behavioral and sensory neuronal response of these mosquito species, in the presence and absence of CO2. When presented alone at ecologically relevant concentrations, acetone increases attraction in Ae. aegypti, but not in An. coluzzii. Moreover, in combination with CO2, human breath-equivalents of acetone ranging between 0.1 and 10 ppm reproduces a behavioral response similar to that observed to human breath in host-seeking Ae. aegypti, but not in An. coluzzii. Acetone does, however, reduce attraction to CO2 in An. coluzzii, when presented at a higher concentration of 10 ppm. We identify the capitate peg A neuron of the maxillary palp of both species as a dual detector of CO2 and acetone. The sensory response to acetone, or binary blends of acetone and CO2, reflects the observed behavioral output in both Ae. aegypti and An. coluzzii. We conclude that host recognition is contextual and dependent on a combination of ecologically relevant odorants at naturally occurring concentrations that are encoded, in this case, by differences in the temporal structure of the neuronal response. This information should be considered when designing synthetic blends for that optimally attract mosquitoes for monitoring and control.


Assuntos
Acetona/farmacologia , Comportamento Animal/efeitos dos fármacos , Dióxido de Carbono/farmacologia , Culicidae/fisiologia , Olfato , Aedes/fisiologia , Animais , Anopheles/fisiologia , Expiração , Comportamento de Busca por Hospedeiro/efeitos dos fármacos , Humanos , Malária/transmissão , Odorantes , Febre Amarela/transmissão
13.
Parasit Vectors ; 12(1): 589, 2019 Dec 16.
Artigo em Inglês | MEDLINE | ID: mdl-31842944

RESUMO

BACKGROUND: Mating swarm segregation in closely related insect species may contribute to reproductive isolation. Visual markers are used for swarm formation; however, it is unknown whether they play a key role in swarm location, species segregation and sex aggregation. METHODS: Using two sympatric closely related species of the Anopheles gambiae complex, An. coluzzii and An. gambiae (s.s.), we investigated in both laboratory and semi-field conditions (i) whether males of the two species use visual markers (black cloths) to locate their swarm; and (ii) whether the presence/absence and size of the marker may differentially affect swarm characteristics. We also investigated whether conspecific virgin females use these markers to join male swarm sites. RESULTS: We showed that males of the two species used visual markers but in different ways: An. coluzzii swarm right above the marker whereas An. gambiae (s.s.) locate their swarm at a constant distance of 76.4 ± 0.6 cm from a 20 × 20 cm marker in the laboratory setup and at 206 ± 6 cm from a 60 × 60 cm marker in the semi-field setup. Although increased marker size recruited more mosquitoes and consequently increased the swarm size in the two species, An. coluzzii swarms flew higher and were stretched both vertically and horizontally, while An. gambiae (s.s.) swarms were only stretched horizontally. Virgin females displayed a swarm-like behavior with similar characteristics to their conspecific males. CONCLUSIONS: Our results provided experimental evidence that both An. coluzzii and An. gambiae (s.s.) males use ground visual markers to form and locate their swarm at species-specific locations. Moreover, the marker size differentially affected swarm characteristics in the two species. Our results also showed that virgin females displayed a swarm-like behavior. However, these "swarms" could be due to the absence of males in our experimental conditions. Nevertheless, the fact that females displayed these "swarms" with the same characteristics as their respective males provided evidence that visual markers are used by the two sexes to join mating spots. Altogether, this suggests that visual markers and the way species and sexes use them could be key cues in species segregation, swarm location and recognition.


Assuntos
Anopheles/fisiologia , Sinais (Psicologia) , Comportamento Sexual Animal , Visão Ocular , Percepção Visual , Animais , Feminino , Masculino
14.
PLoS Genet ; 15(12): e1008440, 2019 12.
Artigo em Inglês | MEDLINE | ID: mdl-31856182

RESUMO

Small laboratory cage trials of non-drive and gene-drive strains of the Asian malaria vector mosquito, Anopheles stephensi, were used to investigate release ratios and other strain properties for their impact on transgene spread during simulated population modification. We evaluated the effects of transgenes on survival, male contributions to next-generation populations, female reproductive success and the impact of accumulation of gene drive-resistant genomic target sites resulting from nonhomologous end-joining (NHEJ) mutagenesis during Cas9, guide RNA-mediated cleavage. Experiments with a non-drive, autosomally-linked malaria-resistance gene cassette showed 'full introduction' (100% of the insects have at least one copy of the transgene) within 8 weeks (≤ 3 generations) following weekly releases of 10:1 transgenic:wild-type males in an overlapping generation trial design. Male release ratios of 1:1 resulted in cages where mosquitoes with at least one copy of the transgene fluctuated around 50%. In comparison, two of three cages in which the malaria-resistance genes were linked to a gene-drive system in an overlapping generation, single 1:1 release reached full introduction in 6-8 generations with a third cage at ~80% within the same time. Release ratios of 0.1:1 failed to establish the transgenes. A non-overlapping generation, single-release trial of the same gene-drive strain resulted in two of three cages reaching 100% introduction within 6-12 generations following a 1:1 transgenic:wild-type male release. Two of three cages with 0.33:1 transgenic:wild-type male single releases achieved full introduction in 13-16 generations. All populations exhibiting full introduction went extinct within three generations due to a significant load on females having disruptions of both copies of the target gene, kynurenine hydroxylase. While repeated releases of high-ratio (10:1) non-drive constructs could achieve full introduction, results from the 1:1 release ratios across all experimental designs favor the use of gene drive, both for efficiency and anticipated cost of the control programs.


Assuntos
Anopheles/fisiologia , Malária/prevenção & controle , Transgenes , Animais , Animais Geneticamente Modificados , Anopheles/genética , Feminino , Genética Populacional , Abrigo para Animais , Malária/genética , Masculino , Mosquitos Vetores/genética , Mosquitos Vetores/fisiologia , Fenótipo , Comportamento Sexual Animal
15.
Malar J ; 18(1): 441, 2019 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-31870365

RESUMO

BACKGROUND: A clear understanding of mosquito biology is fundamental to the control efforts of mosquito-borne diseases such as malaria. Mosquito mark-release-recapture (MMRR) experiments are a popular method of measuring the survival and dispersal of disease vectors; however, examples with African malaria vectors are limited. Ethical and technical difficulties involved in carrying out MMRR studies may have held back research in this area and, therefore, a device that marks mosquitoes as they emerge from breeding sites was developed and evaluated to overcome the problems of MMRR. METHODS: A modified self-marking unit that marks mosquitoes with fluorescent pigment as they emerge from their breeding site was developed based on a previous design for Culex mosquitoes. The self-marking unit was first evaluated under semi-field conditions with laboratory-reared Anopheles arabiensis to determine the marking success and impact on mosquito survival. Subsequently, a field evaluation of MMRR was conducted in Yombo village, Tanzania, to examine the feasibility of the system. RESULTS: During the semi-field evaluation the self-marking units successfully marked 86% of emerging mosquitoes and there was no effect of fluorescent marker on mosquito survival. The unit successfully marked wild male and female Anopheles gambiae sensu lato (s.l.) in sufficiently large numbers to justify its use in MMRR studies. The estimated daily survival probability of An. gambiae s.l. was 0.87 (95% CI 0.69-1.10) and mean dispersal distance was 579 m (95% CI 521-636 m). CONCLUSIONS: This study demonstrates the successful use of a self-marking device in an MMRR study with African malaria vectors. This method may be useful in investigating population structure and dispersal of mosquitoes for deployment and evaluation of future vector control tools, such as gene drive, and to better parameterize mathematical models.


Assuntos
Distribuição Animal , Anopheles/fisiologia , Entomologia/métodos , Controle de Mosquitos/métodos , Mosquitos Vetores/fisiologia , Animais , Feminino , Longevidade , Malária , Masculino , Tanzânia
16.
Malar J ; 18(1): 445, 2019 Dec 27.
Artigo em Inglês | MEDLINE | ID: mdl-31881898

RESUMO

BACKGROUND: Long-lasting insecticidal nets (LLINs) and indoor residual spraying of insecticide (IRS) are widely recommended for the prevention of malaria in endemic regions. Data from human landing catches provide information on the impact of vector control on vector populations. Here, malaria transmission indoors and outdoors, before and after mass deployment of LLINs and IRS in Uganda was compared. METHODS: The study took place in Tororo district, a historically high transmission area where universal LLIN distribution was conducted in November 2013 and May 2017 and 6 rounds of IRS implemented from December 2014 to July 2018. Human landing catches were performed in 8 houses monthly from October 2011 to September 2012 (pre-intervention period) and every 4 weeks from November 2017 to October 2018 (post-intervention period). Mosquitoes were collected outdoors from 18:00 to 22:00 h and indoors from 18:00 to 06:00 h. Female Anopheles were tested for the presence of Plasmodium falciparum sporozoites and species identification performed using gross dissection and polymerase chain reaction (PCR). RESULTS: The interventions were associated with a decline in human biting rate from 19.6 to 2.3 female Anopheles mosquitoes per house per night (p < 0.001) and annual entomological inoculation rate from 129 to 0 infective bites per person per year (p < 0.001). The proportion of mosquitoes collected outdoors increased from 11.6 to 49.4% (p < 0.001). Prior to the interventions the predominant species was Anopheles gambiae sensu stricto (s.s.), which comprised an estimated 76.7% of mosquitoes. Following the interventions, the predominant species was Anopheles arabiensis, which comprised 99.5% of mosquitoes, with almost complete elimination of An. gambiae s.s. (0.5%). CONCLUSIONS: Mass distribution of LLINs and 6 rounds of IRS dramatically decreased vector density and sporozoite rate resulting in a marked reduction in malaria transmission intensity in a historically high transmission site in Uganda. These changes were accompanied by a shift in vector species from An. gambiae s.s. to An. arabiensis and a relative increase in outdoor biting.


Assuntos
Anopheles/fisiologia , Biodiversidade , Mordeduras e Picadas de Insetos/etiologia , Malária/transmissão , Controle de Mosquitos , Mosquitos Vetores/fisiologia , Animais , Feminino , Uganda
17.
Malar J ; 18(1): 357, 2019 Nov 08.
Artigo em Inglês | MEDLINE | ID: mdl-31703736

RESUMO

BACKGROUND: Insecticide resistance is a growing threat to malaria vector control. Ivermectin, either administered to humans or animals, may represent an alternate strategy to reduce resistant mosquito populations. The aim of this study was to assess the residual or delayed effect of administering a single oral dose of ivermectin to humans on the survival, fecundity and fertility of Anopheles arabiensis in Ethiopia. METHODS: Six male volunteers aged 25-40 years (weight range 64-72 kg) were recruited; four of them received a recommended single oral dose of 12 mg ivermectin and the other two individuals were untreated controls. A fully susceptible insectary colony of An. arabiensis was fed on treated and control participants at 1, 4, 7, 10 and 13 days post ivermectin-administration. Daily mosquito mortality was recorded for 5 days. An. arabiensis fecundity and fertility were measured from day 7 post treatment, by dissection to examine the number of eggs per mosquito, and by observing larval hatching rates, respectively. RESULTS: Ivermectin treatment induced significantly higher An. arabiensis mortality on days 1 and 4, compared to untreated controls (p = 0.02 and p < 0.001, respectively). However, this effect had declined by day 7, with no significant difference in mortality between treated and control groups (p = 0.06). The mean survival time of mosquitoes fed on day 1 was 2.1 days, while those fed on day 4 survived 4.0 days. Mosquitoes fed on the treatment group at day 7 and 10 produced significantly lower numbers of eggs compared to the untreated controls (p < 0.001 and p = 0.04, respectively). An. arabiensis fed on day 7 on treated men also had lower larval hatching rates than mosquitoes fed on days 10 and 13 (p = 0.003 and p = 0.001, respectively). CONCLUSION: A single oral dose of ivermectin given to humans can induce mortality and reduce survivorship of An. arabiensis for 7 days after treatment. Ivermectin also had a delayed effect on fecundity of An. arabiensis that took bloodmeals from treated individuals on day 7 and 10. Additional studies are warranted using wild, insecticide-resistant mosquito populations, to confirm findings and a phase III evaluation among community members in Ethiopia is needed to determine the impact of ivermectin on malaria transmission.


Assuntos
Anopheles/efeitos dos fármacos , Antimaláricos/farmacologia , Ivermectina/farmacologia , Adulto , Animais , Anopheles/fisiologia , Etiópia , Fertilidade/efeitos dos fármacos , Humanos , Longevidade/efeitos dos fármacos , Masculino
18.
PLoS Pathog ; 15(11): e1008063, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31697788

RESUMO

Mating causes dramatic changes in female physiology, behaviour, and immunity in many insects, inducing oogenesis, oviposition, and refractoriness to further mating. Females from the Anopheles gambiae species complex typically mate only once in their lifetime during which they receive sperm and seminal fluid proteins as well as a mating plug that contains the steroid hormone 20-hydroxyecdysone. This hormone, which is also induced by blood-feeding, plays a major role in activating vitellogenesis for egg production. Here we show that female Anopheles coluzzii susceptibility to Plasmodium falciparum infection is significantly higher in mated females compared to virgins. We also find that mating status has a major impact on the midgut transcriptome, detectable only under sugar-fed conditions: once females have blood-fed, the transcriptional changes that are induced by mating are likely masked by the widespread effects of blood-feeding on gene expression. To determine whether increased susceptibility to parasites could be driven by the additional 20E that mated females receive from males, we mimicked mating by injecting virgin females with 20E, finding that these females are significantly more susceptible to human malaria parasites than virgin females injected with the control 20E carrier. Further RNAseq was carried out to examine whether the genes that change upon 20E injection in the midgut are similar to those that change upon mating. We find that 79 midgut-expressed genes are regulated in common by both mating and 20E, and 96% (n = 76) of these are regulated in the same direction (up vs down in 20E/mated). Together, these findings show that male Anopheles mosquitoes induce changes in the female midgut that can affect female susceptibility to P. falciparum. This implies that in nature, males might contribute to malaria transmission in previously unappreciated ways, and that vector control strategies that target males may have additional benefits towards reducing transmission.


Assuntos
Anopheles/fisiologia , Sistema Digestório/fisiopatologia , Malária/transmissão , Mosquitos Vetores/patogenicidade , Comportamento Sexual Animal/fisiologia , Transcriptoma , Animais , Sistema Digestório/metabolismo , Sistema Digestório/parasitologia , Suscetibilidade a Doenças , Feminino , Humanos , Hormônios de Inseto/metabolismo , Malária/parasitologia , Masculino , Oviposição , Reprodução
19.
Parasit Vectors ; 12(1): 558, 2019 Nov 26.
Artigo em Inglês | MEDLINE | ID: mdl-31771626

RESUMO

BACKGROUND: The ecology of many mosquitoes, including Anopheles farauti, the dominant malaria vector in the southwest Pacific including the Solomon Islands, remains inadequately understood. Studies to map fine scale vector distributions are biased when trapping techniques use lures that will influence the natural movements of mosquitoes by attracting them to traps. However, passive collection methods allow the detailed natural distributions of vector populations by sex and physiological states to be revealed. METHODS: The barrier screen, a passive mosquito collection method along with human landing catches were used to record An. farauti distributions over time and space in two Solomon Island villages from May 2016 to July 2017. RESULTS: Temporal and spatial distributions of over 15,000 mosquitoes, including males as well as unfed, host seeking, blood-fed, non-blood fed and gravid females were mapped. These spatial and temporal patterns varied by species, sex and physiological state. Sugar-fed An. farauti were mostly collected between 10-20 m away from houses with peak activity from 18:00 to 19:00 h. Male An. farauti were mostly collected greater than 20 m from houses with peak activity from 19:00 to 20:00 h. CONCLUSIONS: Anopheles farauti subpopulations, as defined by physiological state and sex, are heterogeneously distributed in Solomon Island villages. Understanding the basis for these observed heterogeneities will lead to more accurate surveillance of mosquitoes and will enable spatial targeting of interventions for greater efficiency and effectiveness of vector control.


Assuntos
Distribuição Animal , Anopheles/fisiologia , Mosquitos Vetores/fisiologia , Animais , Comportamento Alimentar , Feminino , Habitação , Masculino , Melanesia , Fatores Sexuais , Análise Espaço-Temporal
20.
PLoS One ; 14(11): e0225637, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-31751420

RESUMO

Insects express chemical receptors within sensory neurons that are activated by specific cues in the environment, thereby influencing the acquisition of critical resources. A significant gap in our current understanding of insect chemical ecology is defining the molecular mechanisms that underlie sensitivity to plant-emitted volatiles. Linalool is a commonly-occurring monoterpene that has various effects on insect behavior, either acting as an attractant or a repellent, and existing in nature as one of two possible stereoisomers, (R)-(-)-linalool and (S)-(+)-linalool. In this study, we have used a cell-based functional assay to identify linalool and structurally-related compounds as ligands of Odorant receptor 29, a labellum-expressed receptor in the malaria vector mosquito, Anopheles gambiae (AgamOr29). While (R)-(-)-linalool activates AgamOr29, a mixture of the (R) and (S) stereoisomers activates the receptor with higher potency, implying enantiomeric selectivity. Orthologs of Or29 are present in the genomes of Anophelines within the Cellia subgenus. The conservation of this receptor across Anopheline lineages suggests that this ecologically important compound might serve as an attraction cue for nectar-seeking mosquitoes. Moreover, the characterization of a mosquito terpene receptor could serve as a foundation for future ligand-receptor studies of plant volatiles and for the discovery of compounds that can be integrated into push-pull vector control strategies.


Assuntos
Monoterpenos Acíclicos/farmacologia , Anopheles/fisiologia , Perfilação da Expressão Gênica/métodos , Receptores Odorantes/genética , Animais , Anopheles/efeitos dos fármacos , Comportamento Animal/efeitos dos fármacos , Clonagem Molecular , Regulação da Expressão Gênica/efeitos dos fármacos , Filogenia , Extratos Vegetais/farmacologia , Receptores Odorantes/química , Estereoisomerismo
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