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Chromosomal differentiation and adaptation to human environments in the Anopheles gambiae complex.

Speciation in the Anopheles gambiae complex is reviewed and discussed with emphasis on the patterns of chromosomal differentiation, particularly at the intraspecific level. The significance of inversion polymorphism in gambiae and arabiensis (the two species of greatest medical importance) is evaluated with reference to recent field investigations carried out in Nigeria. In both sibling species some of the inversions show clinical geographical changes in frequencies, with evident correlations with climatic conditions and vegetation zones. Microgeographical variations in species distribution and in intraspecific inversion frequencies are also present, which appear mostly related to man-made environmental contrasts. Parallel indoor-/outdoor collections of samples from polymorphic populations of arabiensis and gambiae show that adult mosquitoes carrying certain inversion karyotypes do not distribute at random in relation to the human environment, being significantly more frequent in outdoor than in indoor samples, or vice-versa. Optimal habitat choice appears to be involved in such variations of indoor resting behaviour, since the chromosomal types carried by less endophilic individuals are those more adapted to humid climates, i.e. those which tend to avoid the higher nocturnal saturation deficit of the indoor environment. This phenomenon, producing non-uniform exposure of the vector population to residual insecticides sprayed in houses, might explain the mediocrity of the results of malaria control projects based on house-spraying against endophilic vectors in the African savannas.

Animals

Plasmodium falciparum sporozoite loads in Anopheles gambiae s.l. across agro-ecological zones of Benin: a cross-sectional field study.

BACKGROUND: Malaria transmission depends critically on the vectorial competence of Anopheles mosquitoes, which is partly reflected by the quantity of infectious Plasmodium falciparum forms harboured in the salivary glands. Whether agro-ecological heterogeneity and vector species identity modulate this parasite burden remains poorly characterized in Benin. METHODS: Anopheles gambiae s.l. were collected by Human Landing Catch across 12 localities spanning 7 agro-ecological zones in Benin during August-October 2022. Species were identified by SINE-PCR. P. falciparum infection was assessed by circumsporozoite protein (CSP) ELISA, and sporozoite load was quantified in positive head-thorax preparations by real-time PCR targeting the plasmepsin gene (NZYTech kit, ref. MD 02411). Statistical comparisons across agro-ecological zones (within each species) used the Kruskal-Wallis test, and the two-species comparison used the Mann-Whitney U test, in R v4.4.1. RESULTS: A total of 1,355 individuals were molecularly analyzed for species identification. Of 3,021 mosquitoes collected, these 1,355 yielded 858 An. coluzzii (63.32%), 489 An. gambiae (36.09%), and 8 An. arabiensis (0.59%). The overall sporozoite index was 5.98% (95% CI 4.8-7.4%), ranging from 0% in Cotonou and Parakou to 16.19% in Boukoumb&#xe9;. No significant inter-zonal variation in sporozoite load was detected in either An. coluzzii (Kruskal-Wallis: H&#x2009;=&#x2009;5.79, df&#x2009;=&#x2009;3, p&#x2009;=&#x2009;0.122) or An. gambiae (H&#x2009;=&#x2009;3.25, df&#x2009;=&#x2009;2, p&#x2009;=&#x2009;0.197). However, An. coluzzii carried a significantly higher sporozoite burden than An. gambiae (Mann-Whitney U&#x2009;=&#x2009;381, p&#x2009;<&#x2009;0.001). CONCLUSIONS: Broad agro-ecological zone categories were not associated with detectable differences in individual P. falciparum sporozoite load in the principal malaria vectors of Benin. Anopheles coluzzii carried a significantly higher sporozoite burden than An. gambiae, indicating species-level differences in parasite permissiveness that merit further investigation.

Anopheles coluzzii

Differentiation between species of the Anopheles gambiae Giles complex (Diptera: Culicidae) by analysis of cuticular hydrocarbons.

Lipids were extracted from 500 female Anopheles gambiae and An. arabiensis in n-hexane and the extracts examined by gas chromatographic analysis using a glass column packed with 3% OV-1 on a 100-200 mesh Gas Chrom Q which was maintained at 225 degrees C. Cuticular hydrocarbons consisted mostly of unbranched paraffins. There were differences between the distribution of compounds with 25-29 carbons. After adjusting the C27 peak to 100%, hydrocarbons C26 and C29 represent heights of 80 and 69% in An. gambiae compared with values of 49 and 95 % in An. arabiensis. Other small but consistent differences were also found. Investigations are being performed on extracts from smaller samples and on other species within the An. gambiae complex.

Animals

Fluorescence banding techniques in the identification of sibling species of the anopheles gambiae complex.

The mitotic chromosomes of the sibling species A and B of the Anopheles gambiae complex were stained with Hoechst 33258 and examined by fluorescence microscopy. The autosomes fluoresce homogenously and similarly in both species while the sex chromosomes differ in the location and brightness of some heterochromatic blocks. These cytochemical differences allow the cytotaxonomic identification of these cryptic species.

Animals

Studies on multiple feeding by Anopheles gambiae s.l. in a Sudan savanna area of north Nigeria.

The detection of haptoglobins in Anopheles gambiae s.l. has been used to obtain an estimate of the incidence of multiple feeding for the village of Barmawa, Garki District, Kano State, Nigeria. The results indicated that the incidence of multiple feeding was approximately 10% but problems were encountered by the high incidence of ahaptoglobinaemia in the population. In four villages in Garki District the incidence of ahaptoglobinaemia varied between 65 and 76% while in young children and personnel under constant malaria chemoprophylaxis it was less than 30%. A strong correlation between ahaptoglobinaemia and malaria infections was seen. The results show evidence of selection of hosts by mosquitoes at Barmawa although this does not necessarily imply a preference per se. The results provide evidence of movement of blood-fed mosquitoes, between houses and from houses to resting sites.

Animals

An ultrastructural study of the sporogonic development of Plasmodium falciparum in Anopheles gambiae.

This paper describes the fine structure of the sporogonic development of Plasmodium falciparum in its natural vector Anopheles gambiae (Species A) as seen by scanning and transmission electron microscopy. The parasite was derived from naturally infected volunteers and the vector maintained under natural conditions at the MRC Laboratories, Fajara, The Gambia. Sporogonic development of P. falciparum is similar to that described for other Plasmodium spp. There are however greater similarities between P. falciparum and the avian malaria parasites, than those mammalian (primarily rodent) species described to date--particularly with respect to mitochondrial development, crystalloid morphology and nucleolar organization. Nuclear development is similar to that of the murine malaria parasites, but reconstruction of complete mitotic spindles from serial sections suggest the haploid genome of P. falciparum contains 14 chromosomes compared to eight to ten in the murine plasmodia. Sporoblast formation involves a unique process of cleft formation based on the expansion of the cisternal space of the endoplasmic reticulum. Sporozoite budding is almost exclusively confined to these inner membrane surfaces and results in a characteristic sporozoite distribution in the oocyst. High resolution scanning electron microscopy of free sporozoites provides the first surface view of the micropore of Plasmodium.

Anopheles

Combination of computational techniques and RNAi reveal targets in Anopheles gambiae for malaria vector control.

Increasing reports of insecticide resistance continue to hamper the gains of vector control strategies in curbing malaria transmission. This makes identifying new insecticide targets or alternative vector control strategies necessary. CLassifier of Essentiality AcRoss EukaRyote (CLEARER), a leave-one-organism-out cross-validation machine learning classifier for essential genes, was used to predict essential genes in Anopheles gambiae and selected predicted genes experimentally validated. The CLEARER algorithm was trained on six model organisms: Caenorhabditis elegans, Drosophila melanogaster, Homo sapiens, Mus musculus, Saccharomyces cerevisiae and Schizosaccharomyces pombe, and employed to identify essential genes in An. gambiae. Of the 10,426 genes in An. gambiae, 1,946 genes (18.7%) were predicted to be Cellular Essential Genes (CEGs), 1716 (16.5%) to be Organism Essential Genes (OEGs), and 852 genes (8.2%) to be essential as both OEGs and CEGs. RNA interference (RNAi) was used to validate the top three highly expressed non-ribosomal predictions as probable vector control targets, by determining the effect of these genes on the survival of An. gambiae G3 mosquitoes. In addition, the effect of knockdown of arginase (AGAP008783) on Plasmodium berghei infection in mosquitoes was evaluated, an enzyme we computationally inferred earlier to be essential based on chokepoint analysis. Arginase and the top three genes, AGAP007406 (Elongation factor 1-alpha, Elf1), AGAP002076 (Heat shock 70kDa protein 1/8, HSP), AGAP009441 (Elongation factor 2, Elf2), had knockdown efficiencies of 91%, 75%, 63%, and 61%, respectively. While knockdown of HSP or Elf2 significantly reduced longevity of the mosquitoes (p<0.0001) compared to control groups, Elf1 or arginase knockdown had no effect on survival. However, arginase knockdown significantly reduced P. berghei oocytes counts in the midgut of mosquitoes when compared to LacZ-injected controls. The study reveals HSP and Elf2 as important contributors to mosquito survival and arginase as important for parasite development, hence placing them as possible targets for vector control.

Animals

Sweeps in Space: Leveraging Geographic Data to Identify Beneficial Alleles in Anopheles gambiae.

As organisms adapt to environmental changes, natural selection modifies the frequency of nonneutral alleles. For beneficial mutations, the outcome of this process may be a selective sweep, in which an allele rapidly increases in frequency and perhaps reaches fixation within a population. Selective sweeps have well-studied effects on patterns of local genetic variation in panmictic populations, but much less is known about the dynamics of sweeps in continuous space. In particular, because limited movement across a landscape leads to unique patterns of population structure, spatial dynamics may influence the trajectory of selected mutations. Here, we use forward-in-time, individual-based simulations in continuous space to study the impact of space on beneficial mutations as they sweep through a population. In particular, we show that selection changes the joint distribution of allele frequency and geographic range occupied by a focal allele and demonstrate that this signal can be used to identify selective sweeps. We then leverage this signal to identify in-progress selective sweeps within the malaria vector Anopheles gambiae, a species under strong selection pressure from vector control measures. By considering space, we identify multiple previously undescribed variants with potential phenotypic consequences, including mutations impacting known IR-associated genes and altering protein structure and properties. Our results demonstrate a novel signal for detecting selection in spatial population genetic data that may have implications for genomic surveillance and understanding geographic patterns of genetic variation.

Animals

Sweeps in space: leveraging geographic data to identify beneficial alleles in Anopheles gambiae.

As organisms adapt to environmental changes, natural selection modifies the frequency of non-neutral alleles. For beneficial mutations, the outcome of this process may be a selective sweep, in which an allele rapidly increases in frequency and perhaps reaches fixation within a population. Selective sweeps have well-studied effects on patterns of local genetic variation in panmictic populations, but much less is known about the dynamics of sweeps in continuous space. In particular, because limited movement across a landscape leads to unique patterns of population structure, spatial dynamics may influence the trajectory of selected mutations. Here, we use forward-in-time, individual-based simulations in continuous space to study the impact of space on beneficial mutations as they sweep through a population. In particular, we show that selection changes the joint distribution of allele frequency and geographic range occupied by a focal allele and demonstrate that this signal can be used to identify selective sweeps. We then leverage this signal to identify in-progress selective sweeps within the malaria vector Anopheles gambiae , a species under strong selection pressure from vector control measures. By considering space, we identify multiple previously undescribed variants with potential phenotypic consequences, including mutations impacting known IR-associated genes and altering protein structure and properties. Our results demonstrate a novel signal for detecting selection in spatial population genetic data that may have implications for genomic surveillance and understanding geographic patterns of genetic variation.

Journal Article

Studies on filariasis transmission in Kwale, a Tanzanian coastal village, and the results of mosquito control measures.

The main vectors of bancroftian filariasis in Kwale, Tanzania, were Anopheles gambiae complex and Culex p. quinquefasciatus. After vector control by the insecticides Dursban (chlorpyrifos) or Abate (temephos), the number of all man-biting mosquitoes was reduced by 94.8%, and that of house-resting mosquitoes by 95.3%. Anopheles gambiae complex were highly exophilic, possibly due to a high proportion of Anopheles merus in this village. The infection rate of A. gambiae complex rose significantly in the post-control samples (P less than 0.05), but nevertheless the overall reduction in filariasis transmission was 87.2%. The infection rate of A. gambiae complex caught off human bait was significantly higher than that of the spray-caught sample (P less than 0.05). It was suggested that the former sample contained a higher proportion of A. merus. This species may be a better vector of filariasis than freshwater forms.

Animals

Chlorfenapyr-pyrethroid nets for pyrethroid-resistant malaria vectors: efficacy, resistance risks, and policy implications.

The Global Technical Strategy for Malaria 2016-2030 aims to reduce malaria incidence and mortality by 90%, yet widespread pyrethroid resistance among major malaria vectors in sub-Saharan Africa threatens this goal. Thus, the World Health Organization recommends chlorfenapyr-pyrethroid combination nets as a priority intervention where pyrethroid resistance undermines vector control. This systematic review synthesizes evidence on the performance, emerging resistance risks, and policy implications of these next-generation insecticide-treated nets. A structured search of literature from 2010 to 2024 across PubMed, Embase, WHO IRIS, and Google Scholar identified 31 eligible studies from 113 records. Evidence shows that chlorfenapyr-pyrethroid nets consistently outperform pyrethroid-only nets against resistant Anopheles populations, demonstrating a 1.8-fold increase in mosquito mortality (95% CI: 1.5-2.1). Community trials report 40-60% reductions in malaria infection incidence and entomological inoculation rates following deployment. However, early signs of chlorfenapyr resistance have emerged in Anopheles gambiae populations in Central Africa (RR: 2.4, p&#x2009;=&#x2009;0.01), linked to CYP6P4 metabolic overexpression. A significant correlation was also observed between agricultural pesticide use and vector resistance patterns (r&#x2009;=&#x2009;0.62, p&#x2009;<&#x2009;0.05). Although chlorfenapyr-pyrethroid nets provide an important short-term tool for managing pyrethroid resistance, their long-term effectiveness depends on integrated resistance management. Rotational deployment with other insecticide classes, strengthened genetic and phenotypic surveillance, and a coordinated 'One Health' approach involving both public health and agriculture are essential to sustain gains and advance progress toward the 2030 malaria targets.

Pyrethrins

AnoEST: toward A. gambiae functional genomics.

Here, we present an analysis of 215,634 EST and cDNA sequences of a major vector of human malaria Anopheles gambiae structured into the AnoEST database. The expressed sequences are grouped into clusters using genomic sequence as template and associated with inferred functional annotation, including the following: corresponding Ensembl gene prediction, putative orthologous genes in other species, homology to known proteins, protein domains, associated Gene Ontology terms, and corresponding classification into broad GO-slim functional groups. AnoEST is a vital resource for interpretation of expression profiles derived using recently developed A. gambiae cDNA microarrays. Using these cDNA microarrays, we have experimentally confirmed the expression of 7961 clusters during mosquito development. Of these, 3100 are not associated with currently predicted genes. Moreover, we found that clusters with confirmed expression are nonbiased with respect to the current gene annotation or homology to known proteins. Consequently, we expect that many as yet unconfirmed clusters are likely to be actual A. gambiae genes. [AnoEST is publicly available at http://komar.embl.de, and is also accessible as a Distributed Annotation Service (DAS).].

Animals