Population subgroups of the African malaria vector Anopheles gambiae have not been comprehensively characterized owing to the lack of unbiased sampling methods. In the arid savanna zone of West Africa, where potential oviposition sites are scarce, widespread collection from larval pools in the peridomestic human habitat yielded a comprehensive genetic survey of local A. gambiae population subgroups, independent of adult resting behavior and ecological preference. A previously unknown subgroup of exophilic A. gambiae is sympatric with the known endophilic A. gambiae in this region. The exophilic subgroup is abundant, lacks differentiation into M and S molecular forms, and is highly susceptible to infection with wild Plasmodium falciparum. These findings might have implications for the epidemiology of malaria transmission and control.Much of the current genetic knowledge of African malaria vector populations is based on collection and analysis of indoor house-resting (endophilic) mosquitoes. Indoor capture of mosquitoes is often regarded as an unbiased collection method for epidemiologically important vectors of human malaria, including A. gambiae sensu stricto (ss), because they are thought to be 'naturally endophilic' (1). Outdoor-resting (exophilic) mosquitoes can contribute to malaria transmission but are underrepresented or absent from indoor collections, particularly if they also bite outdoors (2-7). Collection methods for exophilic mosquitoes are much less efficient than for indoor-resting mosquitoes, involving variants of i) artificial resting sites such as pits, boxes or pots, ii) manual aspiration from vegetation and holes, or iii) capture of mosquitoes landing on animal or human bait, now largely proscribed due to the risk of infection of human collectors (2). Further highlighting the challenge in sampling exophilic mosquitoes, resting sites thought to harbor large A. gambiae populations during the dry season have resisted detection for decades (8)(9)(10) Genetic division of A. gambiae populations into subgroups allows fine ecological partitioning by the species, mediating the expansion of malaria transmission spatially and temporally. Chromosome inversion polymorphisms define certain subgroups (10,12). Frequency of the 2La inversion follows a geographic cline from the humid Central African forest, where the wild-type 2La+ allele is fixed, north to the arid West African savanna, where the inverted 2La allele is fixed (12). One likely phenotype of the 2La inversion is adaptation to aridity. Another example of niche expansion of A. gambiae by genetic subdivision is represented by two genetically diverged molecular forms, termed M and S (13). These molecular forms are detected by assays for fixed nucleotide differences on the X chromosome (Molecular Form Diagnostic SNPs, MFDS), and display other fixed SNPs in genomic 'speciation islands ' (14). The M form dominates in marginal and disturbed habitats where S is less competitive (15).We collected mosquito larvae from natural breeding pools in an arid...

No abstract

The C-terminal proteolytic processing product of merozoite surface protein 1 (MSP1) appears essential for successful erythrocyte invasion by the malarial parasite, Plasmodium. We have determined the crystal structure at 1.8 A resolution of a soluble baculovirus-recombinant form of the protein from P. cynomolgi, which confers excellent protective efficacy in primate vaccination trials. The structure comprises two EGF-like domains, and sequence comparisons strongly suggest that the same conformation is present in all species of Plasmodium, including P. falciparum and P. vivax, which are pathogenic in man. In particular, conserved interdomain contacts between the two EGF modules should preserve the compact form of the molecule in all species. Implications of the crystal structure for anti-malarial vaccine development are discussed.

BackgroundMembers of the Anophelesgambiae species complex are primary vectors of human malaria in Africa. It is known that a large haplotype shared between An. gambiae and Anophelescoluzzii by introgression carries point mutations of the voltage-gated sodium channel gene para, including the L1014F kdr mutation associated with insensitivity to pyrethroid insecticides. Carriage of L1014F kdr is also correlated with higher susceptibility to infection with Plasmodium falciparum. However, the genetic mechanism and causative gene(s) underlying the parasite susceptibility phenotype are not known.MethodsMosquitoes from the wild Burkina Faso population were challenged by feeding on natural P. falciparum gametocytes. Oocyst infection phenotypes were determined and were tested for association with SNP genotypes. Candidate genes in the detected locus were prioritized and RNAi-mediated gene silencing was used to functionally test for gene effects on P. falciparum susceptibility.ResultsA genetic locus, Pfin6, was identified that influences infection levels of P. falciparum in mosquitoes. The locus segregates as a ~3 Mb haplotype carrying 65 predicted genes including the para gene. The haplotype carrying the kdr allele of para is linked to increased parasite infection prevalence, but many single nucleotide polymorphisms on the haplotype are also equally linked to the infection phenotype. Candidate genes in the haplotype were prioritized and functionally tested. Silencing of para did not influence P. falciparum infection, while silencing of a predicted immune gene, serine protease ClipC9, allowed development of significantly increased parasite numbers.ConclusionsGenetic variation influencing Plasmodium infection in wild Anopheles is linked to a natural ~3 megabase haplotype on chromosome 2L that carries the kdr allele of the para gene. Evidence suggests that para gene function does not directly influence parasite susceptibility, and the association of kdr with infection may be due to tight linkage of kdr with other gene(s) on the haplotype. Further work will be required to determine if ClipC9 influences the outcome of P. falciparum infection in nature, as well as to confirm the absence of a direct influence by para.Electronic supplementary materialThe online version of this article (doi:10.1186/s12936-015-0924-8) contains supplementary material, which is available to authorized users.