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1.
Mem Inst Oswaldo Cruz ; 116: e200547, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34076041

RESUMO

BACKGROUND: Forty percent of the world's population live in areas where they are at risk from dengue fever, dengue hemorrhagic fever, and dengue shock syndrome. Dengue viruses are transmitted primarily by the mosquito Aedes aegypti. In Cali, Colombia, approximately 30% of field collected Ae. aegypti are naturally refractory to all four dengue serotypes. OBJECTIVES: Use RNA-sequencing to identify those genes that determine refractoriness in feral mosquitoes to dengue. This information can be used in gene editing strategies to reduce dengue transmission. METHODS: We employed a full factorial design, analyzing differential gene expression across time (24, 36 and 48 h post bloodmeal), feeding treatment (blood or blood + dengue-2) and strain (susceptible or refractory). Sequences were aligned to the reference Ae. aegypti genome for identification, assembled to visualize transcript structure, and analyzed for dynamic gene expression changes. A variety of clustering techniques was used to identify the differentially expressed genes. FINDINGS: We identified a subset of genes that likely assist dengue entry and replication in susceptible mosquitoes and contribute to vector competence. MAIN CONCLUSIONS: The differential expression of specific genes by refractory and susceptible mosquitoes could determine the phenotype, and may be used to in gene editing strategies to reduce dengue transmission.


Assuntos
Aedes , Vírus da Dengue , Dengue , Aedes/genética , Animais , Colômbia , Vírus da Dengue/genética , Mosquitos Vetores/genética , RNA , Transcriptoma/genética
2.
BMC Genomics ; 22(1): 337, 2021 May 10.
Artigo em Inglês | MEDLINE | ID: mdl-33971808

RESUMO

BACKGROUND: Malaria control is heavily reliant on the use of insecticides that target and kill the adult female Anopheline vector. The intensive use of insecticides of the pyrethroid class has led to widespread resistance in mosquito populations. The intensity of pyrethroid resistance in some settings in Africa means mosquitoes can contact bednets treated with this insecticide class multiple times with minimal mortality effects. Furthermore, both ageing and diel cycle have been shown to have large impacts on the resistance phenotype. Together, these traits may affect other aspects of vector biology controlling the vectorial capacity or fitness of the mosquito. RESULTS: Here we show that sublethal exposure of a highly resistant Anopheles coluzzii population originally from Burkina Faso to the pyrethroid deltamethrin results in large and sustained changes to transcript expression. We identify five clear patterns in the data showing changes to transcripts relating to: DNA repair, respiration, translation, behaviour and oxioreductase processes. Further, we highlight differential regulation of transcripts from detoxification families previously linked with insecticide resistance, in addition to clear down-regulation of the oxidative phosphorylation pathway both indicative of changes in metabolism post-exposure. Finally, we show that both ageing and diel cycle have major effects on known insecticide resistance related transcripts. CONCLUSION: Sub-lethal pyrethroid exposure, ageing and the diel cycle results in large-scale changes in the transcriptome of the major malaria vector Anopheles coluzzii. Our data strongly supports further phenotypic studies on how transcriptional changes such as reduced expression of the oxidative phosphorylation pathway or pyrethroid induced changes to redox state might impact key mosquito traits, such as vectorial capacity and life history traits.


Assuntos
Anopheles , Inseticidas , Malária , Piretrinas , Envelhecimento/genética , Animais , Anopheles/genética , Burkina Faso , Feminino , Resistência a Inseticidas/genética , Inseticidas/toxicidade , Controle de Mosquitos , Mosquitos Vetores/genética , Piretrinas/toxicidade , Transcriptoma
3.
BMC Genomics ; 22(1): 396, 2021 May 27.
Artigo em Inglês | MEDLINE | ID: mdl-34044772

RESUMO

BACKGROUND: Transmission of pathogens by vector mosquitoes is intrinsically linked with mosquito's reproductive strategy because anautogenous mosquitoes require vertebrate blood to develop a batch of eggs. Each cycle of egg maturation is tightly linked with the intake of a fresh blood meal for most species. Mosquitoes that acquire pathogens during the first blood feeding can transmit the pathogens to susceptible hosts during subsequent blood feeding and also vertically to the next generation via infected eggs. Large-scale gene-expression changes occur following each blood meal in various tissues, including ovaries. Here we analyzed mosquito ovary transcriptome following a blood meal at three different time points to investigate blood-meal induced changes in gene expression in mosquito ovaries. RESULTS: We collected ovaries from Aedes aegypti that received a sugar meal or a blood meal on days 3, 10 and 20 post blood meal for transcriptome analysis. Over 4000 genes responded differentially following ingestion of a blood meal on day 3, and 660 and 780 genes on days 10 and 20, respectively. Proteins encoded by differentially expressed genes (DEGs) on day 3 include odorant binding proteins (OBPs), defense-specific proteins, and cytochrome P450 detoxification enzymes. In addition, we identified 580 long non-coding RNAs that are differentially expressed at three time points. Gene ontology analysis indicated that genes involved in peptidase activity, oxidoreductase activity, extracellular space, and hydrolase activity, among others were enriched on day 3. Although most of the DEGs returned to the nonsignificant level compared to the sugar-fed mosquito ovaries following oviposition on days 10 and 20, there remained differences in the gene expression pattern in sugar-fed and blood-fed mosquitoes. CONCLUSIONS: Enrichment of OBPs following blood meal ingestion suggests that these genes may have other functions besides being part of the olfactory system. The enrichment of immune-specific genes and cytochrome P450 genes indicates that ovaries become well prepared to protect their germ line from any pathogens that may accompany the blood meal or from environmental contamination during oviposition, and to deal with the detrimental effects of toxic metabolites.


Assuntos
Aedes , Aedes/genética , Animais , Feminino , Expressão Gênica , Mosquitos Vetores/genética , Ovário , Oviposição
4.
Nat Commun ; 12(1): 2960, 2021 05 20.
Artigo em Inglês | MEDLINE | ID: mdl-34017003

RESUMO

Culex mosquitoes are a global vector for multiple human and animal diseases, including West Nile virus, lymphatic filariasis, and avian malaria, posing a constant threat to public health, livestock, companion animals, and endangered birds. While rising insecticide resistance has threatened the control of Culex mosquitoes, advances in CRISPR genome-editing tools have fostered the development of alternative genetic strategies such as gene drive systems to fight disease vectors. However, though gene-drive technology has quickly progressed in other mosquitoes, advances have been lacking in Culex. Here, we develop a Culex-specific Cas9/gRNA expression toolkit and use site-directed homology-based transgenesis to generate and validate a Culex quinquefasciatus Cas9-expressing line. We show that gRNA scaffold variants improve transgenesis efficiency in both Culex quinquefasciatus and Drosophila melanogaster and boost gene-drive performance in the fruit fly. These findings support future technology development to control Culex mosquitoes and provide valuable insight for improving these tools in other species.


Assuntos
Sistemas CRISPR-Cas/genética , Culex/genética , Tecnologia de Impulso Genético/métodos , Controle de Mosquitos/métodos , Mosquitos Vetores/genética , Animais , Animais Geneticamente Modificados , Drosophila melanogaster/genética , Feminino , Resistência a Inseticidas , Masculino , Mutagênese Sítio-Dirigida/métodos , RNA Guia/genética
5.
Pestic Biochem Physiol ; 173: 104772, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33771251

RESUMO

Metabolic resistance driven by multiple P450 genes is worsening insecticide resistance in malaria vectors. However, it remains unclear whether such multiple over-expression imposes an additive fitness cost in the vectors. Here, we showed that two highly over-expressed P450 genes (CYP6P9a and CYP6P9b) combine to impose additive fitness costs in pyrethroid-resistant Anopheles funestus. Genotyping of the CYP6P9b resistance allele in hybrid mosquitoes from a pyrethroid-resistant FUMOZ-R and the susceptible FANG strains revealed that this gene imposes a fitness cost in resistant mosquitoes similar to CYP6P9a. Homozygote susceptible CYP6P9b_S (SS) significantly lay more eggs than the resistant (OR = 2.2, P = 0.04) and with greater hatching rate (p < 0.04). Homozygote resistant larvae CYP6P9b_R (RR) developed significantly slower than homozygote susceptible from L1-L4 (χ2 = 7.2; P = 0.007) with a late pupation observed for RR compared to both heterozygotes and homozygotes susceptible (χ2 = 11.17; P = 0.0008). No difference was observed between genotypes for adult longevity with no change in allele frequency and gene expression across the lifespan. Furthermore, we established that CYP6P9b combines with CYP6P9a to additively exacerbate the fitness cost of pyrethroid resistance with a greater reduction in fecundity/fertility and increased developmental time of double homozygote resistant mosquitoes. Moreover, an increased proportion of double homozygote susceptible individuals was noted over 10 generations in the insecticide-free environment (χ2 = 6.3; P = 0.01) suggesting a reversal to susceptibility in the absence of selection. Such greater fitness cost imposed by multiple P450 genes shows that resistance management strategy based on rotation could help slow the spread of resistance.


Assuntos
Anopheles , Inseticidas , Malária , Piretrinas , Animais , Anopheles/genética , Sistema Enzimático do Citocromo P-450/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Malária/genética , Mosquitos Vetores/genética , Piretrinas/toxicidade
6.
Zootaxa ; 4920(1): zootaxa.4920.1.7, 2021 Jan 27.
Artigo em Inglês | MEDLINE | ID: mdl-33756679

RESUMO

A thorough knowledge of the presence and spatio-temporal distribution patterns of vector species are pivotal to assess the risk of mosquito-borne diseases in Europe. In 2018, a Culex larva was collected during routine monitoring activities to intercept exotic Aedes mosquito species in the port of Antwerp (Kallo, Belgium). The larva, collected from a pond in mid-September, was morphologically identified as Culex modestus, and this identification was subsequently confirmed by COI barcoding. It is the first confirmed record of this West Nile virus bridge vector in Belgium. The present study also demonstrates the value of DNA-based identification techniques to validate the presence of potential vector species.


Assuntos
Culex , Culicidae , Vírus do Nilo Ocidental , Animais , Bélgica , Culex/genética , DNA , Código de Barras de DNA Taxonômico , Mosquitos Vetores/genética , Vírus do Nilo Ocidental/genética
7.
Biomed Res Int ; 2021: 6649038, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33763480

RESUMO

Background: Genetic modification offers opportunities to introduce artificially created molecular defence mechanisms to vector mosquitoes to counter diseases causing pathogens such as the dengue virus, malaria parasite, and Zika virus. RNA interference is such a molecular defence mechanism that could be used for this purpose to block the transmission of pathogens among human and animal populations. In our previous study, we engineered a dengue-resistant transgenic Ae. aegypti using RNAi to turn off the expression of dengue virus serotype genomes to reduce virus transmission, requiring assessment of the fitness of this mosquito with respect to its wild counterpart in the laboratory and semifield conditions. Method: Developmental and reproductive fitness parameters of TM and WM have assessed under the Arthropod Containment Level 2 conditions, and the antibiotic treatment assays were conducted using co-trimoxazole, amoxicillin, and doxycycline to assess the developmental and reproductive fitness parameters. Results: A significant reduction of developmental and reproductive fitness parameters was observed in transgenic mosquito compared to wild mosquitoes. However, it was seen in laboratory-scale studies that the fitness of this mosquito has improved significantly in the presence of antibiotics such as co-trimoxazole, amoxicillin, and doxycycline in their feed. Conclusion: Our data indicate that the transgenic mosquito produced had a reduction of the fitness parameters and it may lead to a subsequent reduction of transgenic vector density over the generations in field applications. However, antibiotics of co-trimoxazole, amoxicillin, and doxycycline have shown the improvement of fitness parameters indicating the usefulness in field release of transgenic mosquitoes.


Assuntos
Animais Geneticamente Modificados , Antibacterianos/farmacologia , Vírus da Dengue/fisiologia , Aptidão Genética , Mosquitos Vetores , Replicação Viral , Aedes/genética , Aedes/virologia , Animais , Dengue/genética , Dengue/prevenção & controle , Dengue/transmissão , Mosquitos Vetores/genética , Mosquitos Vetores/virologia , Replicação Viral/efeitos dos fármacos , Replicação Viral/genética
8.
Pestic Biochem Physiol ; 173: 104783, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33771262

RESUMO

In the central western Senegal, malaria transmission has been reduced low due to the combination of several effective control interventions. However, despite this encouraging achievement, residual malaria transmission still occurring in few areas, mainly ensured by An. arabiensis and An. melas. The resurgence or the persistence of the disease may have originated from the increase and the spread of insecticide resistance genes among natural malaria vectors populations. Therefore, assessing the status and mechanisms of insecticides resistance among targeted malaria vectors is of highest importance to better characterize factors underlying the residual transmission where it occurs. Malaria vectors were collected from three selected villages using nocturnal human landing catches (HLC) and pyrethrum spray collections (PSC) methods. An. gambiae s.l. specimens were identified at the species level then genotyped for the presence of kdr-west (L1014F), kdr-east (L1014S) and ace-1R mutations by qPCR. An. arabiensis (69.36%) and An. melas (27.99%) were the most common species of the Gambiae complex in the study area. Among An. arabiensis population, the allelic frequency of the kdr-east (22.66%) was relatively higher than for kdr-west mutation (9.96%). While for An. melas populations, the overall frequencies of both mutations were very low, being respectively 1.12% and 0.40% for the L1014S and L1014F mutations. With a global frequency of 2%, only the heterozygous form of the G119S mutation was found only in An. arabiensis and in all the study sites. The widespread occurrence of the kdr mutation in both An. arabiensis and An. melas natural populations, respectively the main and focal vectors in the central-western Senegal, may have contributed to maintaining malaria transmission in the area. Thus, compromising the effectiveness of pyrethroids-based vector control measures and the National Elimination Goal. Therefore, monitoring and managing properly insecticide resistance became a key programmatic intervention to achieve the elimination goal where feasible, as aimed by Senegal. Noteworthy, this is the first report of the ace-1 mutation in natural populations of An. arabiensis from Senegal, which need to be closely monitored to preserve one of the essential insecticide classes used in IRS to control the pyrethroids-resistant populations.


Assuntos
Anopheles , Inseticidas , Malária , Piretrinas , Animais , Anopheles/genética , Resistência a Inseticidas/genética , Inseticidas/farmacologia , Malária/genética , Mosquitos Vetores/genética , Mutação , Piretrinas/farmacologia , Senegal
9.
J Vis Exp ; (168)2021 02 02.
Artigo em Inglês | MEDLINE | ID: mdl-33616090

RESUMO

Functional genomic analysis and related strategies for genetic control of malaria rely on validated and reproducible methods to accurately modify the genome of Anopheles mosquitoes. Amongst these methods, the φC31 system allows precise and stable site-directed integration of transgenes, or the substitution of integrated transgenic cassettes via recombinase-mediated cassette exchange (RMCE). This method relies on the action of the Streptomyces φC31 bacteriophage integrase to catalyze recombination between two specific attachment sites designated attP (derived from the phage) and attB (derived from the host bacterium). The system uses one or two attP sites that have been integrated previously into the mosquito genome and attB site(s) in the donor template DNA. Here we illustrate how to stably modify the genome of attP-bearing Anopheles docking lines using two plasmids: an attB-tagged donor carrying the integration or exchange template and a helper plasmid encoding the φC31 integrase. We report two representative results of φC31-mediated site-directed modification: the single integration of a transgenic cassette in An. stephensi and RMCE in An. gambiae mosquitoes. φC31-mediated genome manipulation offers the advantage of reproducible transgene expression from validated, fitness neutral genomic sites, allowing comparative qualitative and quantitative analyses of phenotypes. The site-directed nature of the integration also substantially simplifies the validation of the single insertion site and the mating scheme to obtain a stable transgenic line. These and other characteristics make the φC31 system an essential component of the genetic toolkit for the transgenic manipulation of malaria mosquitoes and other insect vectors.


Assuntos
Anopheles/genética , Regulação da Expressão Gênica , Integrases/genética , Mosquitos Vetores/genética , Recombinação Genética , Siphoviridae/enzimologia , Transgenes/fisiologia , Animais , Marcação de Genes , Genoma , Malária/transmissão , Mutagênese Sítio-Dirigida , Mutação , Siphoviridae/genética
10.
Cryobiology ; 99: 1-10, 2021 04.
Artigo em Inglês | MEDLINE | ID: mdl-33556359

RESUMO

Mosquito-borne diseases are responsible for millions of human deaths every year, posing a massive burden on global public health. Mosquitoes transmit a variety of bacteria, parasites and viruses. Mosquito control efforts such as insecticide spraying can reduce mosquito populations, but they must be sustained in order to have long term impacts, can result in the evolution of insecticide resistance, are costly, and can have adverse human and environmental effects. Technological advances have allowed genetic manipulation of mosquitoes, including generation of those that are still susceptible to insecticides, which has greatly increased the number of mosquito strains and lines available to the scientific research community. This generates an associated challenge, because rearing and maintaining unique mosquito lines requires time, money and facilities, and long-term maintenance can lead to adaptation to specific laboratory conditions, resulting in mosquito lines that are distinct from their wild-type counterparts. Additionally, continuous rearing of transgenic lines can lead to loss of genetic markers, genes and/or phenotypes. Cryopreservation of valuable mosquito lines could help circumvent these limitations and allow researchers to reduce the cost of rearing multiple lines simultaneously, maintain low passage number transgenic mosquitoes, and bank lines not currently being used. Additionally, mosquito cryopreservation could allow researchers to access the same mosquito lines, limiting the impact of unique laboratory or field conditions. Successful cryopreservation of mosquitoes would expand the field of mosquito research and could ultimately lead to advances that would reduce the burden of mosquito-borne diseases, possibly through rear-and-release strategies to overcome mosquito insecticide resistance. Cryopreservation techniques have been developed for some insect groups, including but not limited to fruit flies, silkworms and other moth species, and honeybees. Recent advances within the cryopreservation field, along with success with other insects suggest that cryopreservation of mosquitoes may be a feasible method for preserving valuable scientific and public health resources. In this review, we will provide an overview of basic mosquito biology, the current state of and advances within insect cryopreservation, and a proposed approach toward cryopreservation of Anopheles stephensi mosquitoes.


Assuntos
Anopheles , Mosquitos Vetores , Animais , Abelhas , Criopreservação/métodos , Humanos , Resistência a Inseticidas/genética , Controle de Mosquitos , Mosquitos Vetores/genética
11.
PLoS Negl Trop Dis ; 15(1): e0008351, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33481791

RESUMO

The Asian tiger mosquito, Aedes albopictus (Ae. albopictus), is an important vector that transmits arboviruses such as dengue (DENV), Zika (ZIKV) and Chikungunya virus (CHIKV). Long noncoding RNAs (lncRNAs) are known to regulate various biological processes. Knowledge on Ae. albopictus lncRNAs and their functional role in virus-host interactions are still limited. Here, we identified and characterized the lncRNAs in the genome of an arbovirus vector, Ae. albopictus, and evaluated their potential involvement in DENV and ZIKV infection. We used 148 public datasets, and identified a total of 10, 867 novel lncRNA transcripts, of which 5,809, 4,139, and 919 were intergenic, intronic and antisense respectively. The Ae. albopictus lncRNAs shared many characteristics with other species such as short length, low GC content, and low sequence conservation. RNA-sequencing of Ae. albopictus cells infected with DENV and ZIKV showed that the expression of lncRNAs was altered upon virus infection. Target prediction analysis revealed that Ae. albopictus lncRNAs may regulate the expression of genes involved in immunity and other metabolic and cellular processes. To verify the role of lncRNAs in virus infection, we generated mutations in lncRNA loci using CRISPR-Cas9, and discovered that two lncRNA loci mutations, namely XLOC_029733 (novel lncRNA transcript id: lncRNA_27639.2) and LOC115270134 (known lncRNA transcript id: XR_003899061.1) resulted in enhancement of DENV and ZIKV replication. The results presented here provide an important foundation for future studies of lncRNAs and their relationship with virus infection in Ae. albopictus.


Assuntos
Aedes/genética , Aedes/virologia , Vírus da Dengue/fisiologia , RNA Longo não Codificante/genética , RNA Longo não Codificante/metabolismo , Zika virus/fisiologia , Aedes/metabolismo , Animais , Sistemas CRISPR-Cas , Linhagem Celular , Dengue/virologia , Vírus da Dengue/genética , Regulação da Expressão Gênica , Genoma , Interações entre Hospedeiro e Microrganismos/genética , Interações entre Hospedeiro e Microrganismos/fisiologia , Mosquitos Vetores/genética , Mosquitos Vetores/virologia , Transcriptoma , Zika virus/genética , Infecção por Zika virus/virologia
12.
PLoS Negl Trop Dis ; 15(1): e0009036, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33497375

RESUMO

BACKGROUND: In the absence of vaccines or drugs, insecticides are the mainstay of Aedes-borne disease control. Their utility is challenged by the slow deployment of resources, poor community compliance and inadequate household coverage. Novel application methods are required. METHODOLOGY AND PRINCIPAL FINDINGS: A 10% w/w metofluthrin "emanator" that passively disseminates insecticide from an impregnated net was evaluated in a randomized trial of 200 houses in Mexico. The devices were introduced at a rate of 1 per room and replaced at 3-week intervals. During each of 7 consecutive deployment cycles, indoor resting mosquitoes were sampled using aspirator collections. Assessments of mosquito landing behaviours were made in a subset of houses. Pre-treatment, there were no differences in Aedes aegypti indices between houses recruited to the control and treatment arms. Immediately after metofluthrin deployment, the entomological indices between the trial arms diverged. Averaged across the trial, there were significant reductions in Abundance Rate Ratios for total Ae. aegypti, female abundance and females that contained blood meals (2.5, 2.4 and 2.3-times fewer mosquitoes respectively; P<0.001). Average efficacy was 60.2% for total adults, 58.3% for females, and 57.2% for blood-fed females. The emanators also reduced mosquito landings by 90% from 12.5 to 1.2 per 10-minute sampling period (P<0.05). Homozygous forms of the pyrethroid resistant kdr alleles V410L, V1016L and F1534C were common in the target mosquito population; found in 39%, 24% and 95% of mosquitoes collected during the trial. CONCLUSIONS/SIGNIFICANCE: This is the first randomized control trial to evaluate the entomological impact of any volatile pyrethroid on urban Ae. aegypti. It demonstrates that volatile pyrethroids can have a sustained impact on Ae. aegypti population densities and human-vector contact indoors. These effects occur despite the presence of pyrethroid-resistant alleles in the target population. Formulations like these may have considerable utility for public health vector control responses.


Assuntos
Aedes/efeitos dos fármacos , Ciclopropanos/farmacologia , Fluorbenzenos/farmacologia , Inseticidas/farmacologia , Controle de Mosquitos/métodos , Mosquitos Vetores/efeitos dos fármacos , Aedes/genética , Animais , Comportamento Animal , Dengue/transmissão , Entomologia , Características da Família , Feminino , Humanos , Resistência a Inseticidas/efeitos dos fármacos , Resistência a Inseticidas/genética , México , Mosquitos Vetores/genética , Prevalência , Piretrinas/farmacologia , Projetos de Pesquisa
13.
PLoS Genet ; 17(1): e1009253, 2021 01.
Artigo em Inglês | MEDLINE | ID: mdl-33476334

RESUMO

Vector population control using insecticides is a key element of current strategies to prevent malaria transmission in Africa. The introduction of effective insecticides, such as the organophosphate pirimiphos-methyl, is essential to overcome the recurrent emergence of resistance driven by the highly diverse Anopheles genomes. Here, we use a population genomic approach to investigate the basis of pirimiphos-methyl resistance in the major malaria vectors Anopheles gambiae and A. coluzzii. A combination of copy number variation and a single non-synonymous substitution in the acetylcholinesterase gene, Ace1, provides the key resistance diagnostic in an A. coluzzii population from Côte d'Ivoire that we used for sequence-based association mapping, with replication in other West African populations. The Ace1 substitution and duplications occur on a unique resistance haplotype that evolved in A. gambiae and introgressed into A. coluzzii, and is now common in West Africa primarily due to selection imposed by other organophosphate or carbamate insecticides. Our findings highlight the predictive value of this complex resistance haplotype for phenotypic resistance and clarify its evolutionary history, providing tools to for molecular surveillance of the current and future effectiveness of pirimiphos-methyl based interventions.


Assuntos
Acetilcolinesterase/genética , Resistência a Inseticidas/genética , Malária/genética , Malária/transmissão , África Ocidental , Animais , Anopheles/efeitos dos fármacos , Anopheles/genética , Anopheles/parasitologia , Variações do Número de Cópias de DNA/genética , Genes Duplicados/genética , Introgressão Genética/genética , Humanos , Inseticidas/efeitos adversos , Malária/parasitologia , Malária/prevenção & controle , Mosquitos Vetores/genética , Compostos Organotiofosforados/efeitos adversos , Compostos Organotiofosforados/farmacologia
14.
Parasit Vectors ; 14(1): 12, 2021 Jan 06.
Artigo em Inglês | MEDLINE | ID: mdl-33407824

RESUMO

BACKGROUND: Aedes albopictus is an indigenous primary vector for dengue and Zika viruses in China. Compared with its insecticide resistance, biology and vector competence, little is known about its genetic variation, which corresponds to environmental variations. Thus, the present study examines how Ae. albopictus varies among different climatic regions in China and deciphers its potential dispersal patterns. METHODS: The genetic variation and population structure of 17 Ae. albopictus populations collected from three climatic regions of China were investigated with 11 microsatellite loci and the mitochondrial coxI gene. RESULTS: Of 44 isolated microsatellite markers, 11 pairs were chosen for genotyping analysis and had an average PIC value of 0.713, representing high polymorphism. The number of alleles was high in each population, with the ne value increasing from the temperate region (3.876) to the tropical region (4.144). Twenty-five coxI haplotypes were detected, and the highest diversity was observed in the tropical region. The mean Ho value (ca. 0.557) of all the regions was significantly lower than the mean He value (ca. 0.684), with nearly all populations significantly departing from HWE and displaying significant population expansion (p value < 0.05). Two genetically isolated groups and three haplotype clades were evaluated via STRUCTURE and haplotype phylogenetic analyses, and the tropical populations were significantly isolated from those in the other regions. Most genetic variation in Ae. albopictus was detected within populations and individuals at 31.40 and 63.04%, respectively, via the AMOVA test, and a relatively significant positive correlation was observed among only the temperate populations via IBD analysis (R2 = 0.6614, p = 0.048). Recent dispersions were observed among different Ae. albopictus populations, and four major migration trends with high gene flow (Nm > 0.4) were reconstructed between the tropical region and the other two regions. Environmental factors, especially temperature and rainfall, may be the leading causes of genetic diversity in different climatic regions. CONCLUSIONS: Continuous dispersion contributes to the genetic communication of Ae. albopictus populations across different climatic regions, and environmental factors, especially temperature and rainfall, may be the leading causes of genetic variation.


Assuntos
Aedes/genética , Distribuição Animal , Genética Populacional , Aedes/virologia , Animais , China/epidemiologia , Clima , Dengue/transmissão , Complexo IV da Cadeia de Transporte de Elétrons/genética , Genes de Insetos , Variação Genética , Repetições de Microssatélites/genética , Mosquitos Vetores/genética , Mosquitos Vetores/virologia , Filogenia , Polimorfismo Genético , Infecção por Zika virus/transmissão
15.
Acta Trop ; 216: 105820, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33400915

RESUMO

Malaria vector control in Mali relies heavily on the use of long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) in selected districts. As part of strengthening vector control strategies in Koulikoro district, the National Malaria Control Programme (NMCP) through the support from the US President's Malaria Initiative (PMI) has strategically driven the implementation of IRS, with the LLINs coverage also rising from 93.3% and 98.2%. Due to the increased reports of vector resistance to both pyrethroid and carbamates, there was a campaign for the use of pirimiphos-methyl, an organophosphate at Koulikoro between 2015 and 2016. In this study, the effect of IRS on malaria transmission was assessed, by comparing some key entomological indices between Koulikoro, where IRS was implemented and its neighboring district, Banamba that has never received IRS as vector control intervention. The study was conducted in two villages of each district (Koulikoro and Banamba). Pyrethrum spray catches and entry window trapping were used to collect mosquitoes on a monthly basis. WHO tube tests were carried out to assess mosquito susceptibility to insecticides. Mosquitoes were identified to species level by PCR and their infection to P. falciparum was detected by Enzyme Linked-Immuno-Sorbent Assay (ELISA). Of the 527 specimens identified, An. coluzzii was the most frequent species (95%) followed by An. gambiae (4%) and An. arabiensis (1%). Its density was rainfall dependent in the no-IRS area, and almost independent in the IRS area. The infection rate (IR) in the no-IRS area was 0.96%, while it was null in the IRS area. In the no-IRS area, the entomological inoculation rate (EIR) was 0.21 infective bites /person month with a peak in September. High resistance to pyrethroids and carbamates and susceptibility to organophosphates was observed at all sites. The introduction of pirimiphos-methyl based IRS for vector control resulted in a significant decrease in malaria transmission. An. gambiae s.l., the main malaria vector in the area, was resistant to pyrethroids and carbamates but remained susceptible to the organophosphate pirimiphos-methyl.


Assuntos
Anopheles/efeitos dos fármacos , Anopheles/parasitologia , Carbamatos/farmacologia , Malária/prevenção & controle , Malária/transmissão , Compostos Organotiofosforados/farmacologia , Piretrinas/farmacologia , Animais , Anopheles/genética , Vetores de Doenças , Feminino , Humanos , Mordeduras e Picadas de Insetos/parasitologia , Resistência a Inseticidas , Inseticidas/farmacologia , Mali , Tipagem Molecular , Controle de Mosquitos/métodos , Mosquitos Vetores/efeitos dos fármacos , Mosquitos Vetores/genética , Reação em Cadeia da Polimerase
16.
BMC Genomics ; 22(1): 71, 2021 Jan 21.
Artigo em Inglês | MEDLINE | ID: mdl-33478394

RESUMO

BACKGROUND: In the light of dengue being the fastest growing transmissible disease, there is a dire need to identify the mechanisms regulating the behaviour of the main vector Aedes aegypti. Disease transmission requires the female mosquito to acquire the pathogen from a blood meal during one gonotrophic cycle, and to pass it on in the next, and the capacity of the vector to maintain the disease relies on a sustained mosquito population. RESULTS: Using a comprehensive transcriptomic approach, we provide insight into the regulation of the odour-mediated host- and oviposition-seeking behaviours throughout the first gonotrophic cycle. We provide clear evidence that the age and state of the female affects antennal transcription differentially. Notably, the temporal- and state-dependent patterns of differential transcript abundance of chemosensory and neuromodulatory genes extends across families, and appears to be linked to concerted differential modulation by subsets of transcription factors. CONCLUSIONS: By identifying these regulatory pathways, we provide a substrate for future studies targeting subsets of genes across disparate families involved in generating key vector behaviours, with the goal to develop novel vector control tools.


Assuntos
Aedes , Dengue , Aedes/genética , Animais , Dengue/genética , Feminino , Humanos , Insetos Vetores/genética , Mosquitos Vetores/genética , Transcriptoma
17.
Acta Trop ; 215: 105799, 2021 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-33358735

RESUMO

Anopheles tessellatus is a potentially important vector found across South, East and Southeast Asia. While it was formerly considered a formidable vector of human Plasmodium and filarial parasites in the Maldives, and of lesser importance as a vector of human Plasmodium in Sri Lanka and parts of Indonesia, it is currently of little or unknown health importance in many other parts of its range. This study describes the genetic diversity and evolutionary relationships among An. tessellatus populations in nine Asian countries at the COI gene using maximum-likelihood and Bayesian phylogenetic inference tree and cluster-based species delimitation approaches. These analyses reveal exceptional levels of genetic diversity in An. tessellatus populations across its known range, and identify up to six putative species in the newly determined Tessellatus Complex. The existence of such cryptic diversity has potentially important consequences for vector management and disease control. Differences in the ecologies and life histories among these species may have considerable impact on vectorial capacity and may go some way towards explaining why An. tessellatus s.l. has such varying degrees of public health importance across its range.


Assuntos
Anopheles/genética , Mosquitos Vetores/genética , Animais , Teorema de Bayes , Variação Genética , Humanos , Malária/transmissão , Filogenia
18.
PLoS Negl Trop Dis ; 14(12): e0008971, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33338046

RESUMO

Aedes aegypti is a vector of dengue, chikungunya, and Zika viruses. Current vector control strategies such as community engagement, source reduction, and insecticides have not been sufficient to prevent viral outbreaks. Thus, interest in novel strategies involving genetic engineering is growing. Female mosquitoes rely on flight to mate with males and obtain a bloodmeal from a host. We hypothesized that knockout of genes specifically expressed in female mosquitoes associated with the indirect flight muscles would result in a flightless female mosquito. Using CRISPR-Cas9 we generated loss-of-function mutations in several genes hypothesized to control flight in mosquitoes, including actin (AeAct-4) and myosin (myo-fem) genes expressed specifically in the female flight muscle. Genetic knockout of these genes resulted in 100% flightless females, with homozygous males able to fly, mate, and produce offspring, albeit at a reduced rate when compared to wild type males. Interestingly, we found that while AeAct-4 was haplosufficient, with most heterozygous individuals capable of flight, this was not the case for myo-fem, where about half of individuals carrying only one intact copy could not fly. These findings lay the groundwork for developing novel mechanisms of controlling Ae. aegypti populations, and our results suggest that this mechanism could be applicable to other vector species of mosquito.


Assuntos
Aedes/genética , Sistemas CRISPR-Cas , Inseticidas/farmacologia , Controle de Mosquitos , Mosquitos Vetores/genética , Infecção por Zika virus/prevenção & controle , Zika virus/fisiologia , Aedes/fisiologia , Aedes/virologia , Animais , Feminino , Voo Animal , Técnicas de Inativação de Genes , Humanos , Masculino , Mosquitos Vetores/fisiologia , Mosquitos Vetores/virologia , Fenótipo , Infecção por Zika virus/transmissão , Infecção por Zika virus/virologia
19.
PLoS Pathog ; 16(12): e1008908, 2020 12.
Artigo em Inglês | MEDLINE | ID: mdl-33347501

RESUMO

Anopheles mosquitoes have transmitted Plasmodium parasites for millions of years, yet it remains unclear whether they suffer fitness costs to infection. Here we report that the fecundity of virgin and mated females of two important vectors-Anopheles gambiae and Anopheles stephensi-is not affected by infection with Plasmodium falciparum, demonstrating that these human malaria parasites do not inflict this reproductive cost on their natural mosquito hosts. Additionally, parasite development is not impacted by mating status. However, in field studies using different P. falciparum isolates in Anopheles coluzzii, we find that Mating-Induced Stimulator of Oogenesis (MISO), a female reproductive gene strongly induced after mating by the sexual transfer of the steroid hormone 20-hydroxyecdysone (20E), protects females from incurring fecundity costs to infection. MISO-silenced females produce fewer eggs as they become increasingly infected with P. falciparum, while parasite development is not impacted by this gene silencing. Interestingly, previous work had shown that sexual transfer of 20E has specifically evolved in Cellia species of the Anopheles genus, driving the co-adaptation of MISO. Our data therefore suggest that evolution of male-female sexual interactions may have promoted Anopheles tolerance to P. falciparum infection in the Cellia subgenus, which comprises the most important malaria vectors.


Assuntos
Anopheles/genética , Interações Hospedeiro-Parasita/genética , Plasmodium falciparum/genética , Animais , Anopheles/parasitologia , Ecdisterona/genética , Ecdisterona/metabolismo , Feminino , Fertilidade/genética , Expressão Gênica , Hormônios/fisiologia , Malária/parasitologia , Malária Falciparum/parasitologia , Masculino , Mosquitos Vetores/genética , Oogênese , Plasmodium falciparum/patogenicidade , Reprodução/fisiologia
20.
BMC Genet ; 21(Suppl 2): 142, 2020 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-33339503

RESUMO

BACKGROUND: Aedes aegypti is the primary vector of arthropod-borne viruses and one of the most widespread and invasive mosquito species. Due to the lack of efficient specific drugs or vaccination strategies, vector population control methods, such as the sterile insect technique, are receiving renewed interest. However, availability of a reliable genetic sexing strategy is crucial, since there is almost zero tolerance for accidentally released females. Development of genetic sexing strains through classical genetics is hindered by genetic recombination that is not suppressed in males as is the case in many Diptera. Isolation of naturally-occurring or irradiation-induced inversions can enhance the genetic stability of genetic sexing strains developed through genetically linking desirable phenotypes with the male determining region. RESULTS: For the induction and isolation of inversions through irradiation, 200 male pupae of the 'BRA' wild type strain were irradiated at 30 Gy and 100 isomale lines were set up by crossing with homozygous 'red-eye' (re) mutant females. Recombination between re and the M locus and the white (w) gene (causing a recessive white eye phenotype when mutated) and the M locus was tested in 45 and 32 lines, respectively. One inversion (Inv35) reduced recombination between both re and the M locus, and wand the M locus, consistent with the presence of a rather extended inversion between the two morphological mutations, that includes the M locus. Another inversion (Inv5) reduced recombination only between w and the M locus. In search of naturally-occurring, recombination-suppressing inversions, homozygous females from the red eye and the white eye strains were crossed with seventeen and fourteen wild type strains collected worldwide, representing either recently colonized or long-established laboratory populations. Despite evidence of varying frequencies of recombination, no combination led to the elimination or substantial reduction of recombination. CONCLUSION: Inducing inversions through irradiation is a feasible strategy to isolate recombination suppressors either on the M or the m chromosome for Aedes aegypti. Such inversions can be incorporated in genetic sexing strains developed through classical genetics to enhance their genetic stability and support SIT or other approaches that aim to population suppression through male-delivered sterility.


Assuntos
Aedes/genética , Aedes/efeitos da radiação , Infertilidade/genética , Recombinação Genética/efeitos da radiação , Animais , Feminino , Raios gama , Genes de Insetos , Marcadores Genéticos , Controle de Insetos , Masculino , Mosquitos Vetores/genética , Mosquitos Vetores/efeitos da radiação
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