Diurnal biting of malaria mosquitoes in the Central African Republic indicates residual transmission may be "out of control".

Malaria control interventions target nocturnal feeding of the Anopheles vectors indoors to reduce parasite transmission. Mass deployment of insecticidal bed nets and indoor residual spraying with insecticides, however, may induce mosquitoes to blood-feed at places and at times when humans are not protected. These changes can set a ceiling to the efficacy of these control interventions, resulting in residual malaria transmission. Despite its relevance for disease transmission, the daily rhythmicity of Anopheles biting behavior is poorly documented, most investigations focusing on crepuscular hours and nighttime. By performing mosquito collections 48-h around the clock, both indoors and outdoors, and by modeling biting events using circular statistics, we evaluated the full daily rhythmicity of biting in urban Bangui, Central African Republic. While the bulk of biting by Anopheles gambiae, Anopheles coluzzii, Anopheles funestus, and Anopheles pharoensis occurred from sunset to sunrise outdoors, unexpectedly ∼20 to 30% of indoor biting occurred during daytime. As biting events did not fully conform to any family of circular distributions, we fitted mixtures of von Mises distributions and found that observations were consistent with three compartments, corresponding indoors to populations of early-night, late-night, and daytime-biting events. It is not known whether these populations of biting events correspond to spatiotemporal heterogeneities or also to distinct mosquito genotypes/phenotypes belonging consistently to each compartment. Prevalence of Plasmodium falciparum in nighttime- and daytime-biting mosquitoes was the same. As >50% of biting occurs in Bangui when people are unprotected, malaria control interventions outside the domiciliary environment should be envisaged.

Host preference patterns in domestic and wild settings: Insights into Anopheles feeding behavior.

The adaptation of Anopheles malaria vectors to domestic settings is directly linked to their ability to feed on humans. The strength of this species-habitat association is unequal across the species within the genus, with the major vectors being particularly dependent on humans. However, our understanding of how blood-feeding behavior interacts with and adapts to environmental settings, including the presence of humans, remains limited. Using a field-based approach, we first investigated Anopheles community structure and feeding behavior patterns in domestic and sylvatic settings in La Lopé National Park in Gabon, Central Africa. We characterized the preference indices using a dual-host choice sampling approach across mosquito species, habitats, and seasons. We then quantified the plastic biting behavior of mosquito species in each habitat. We collected individuals from 16 Anopheles species that exhibited significant differences in species composition and abundance between sylvatic and domestic settings. The host-seeking behavior also varied among the seven most abundant species. The general attractiveness to each host, human or animal, remained relatively constant for each species, but with significant variations between habitats across species. These variations, to more generalist and to more anthropophilic behavior, were related to seasonal changes and distance from the village, respectively. Finally, we pointed out that the host choice of major malaria vectors changed in the absence of humans, revealing a plastic feeding behavior of these species. This study highlights the effect of humans on Anopheles distribution and feeding evolution. The characterization of feeding behavior in wild and domestic settings provides opportunities to better understand the interplay between genetic determinants of host preference and ecological factors. Our findings suggest that protected areas may offer alternative thriving conditions to major malaria vectors.

Climate-influenced vector-borne diseases in Africa: a call to empower the next generation of African researchers for sustainable solutions.

We look at the link between climate change and vector-borne diseases in low- and middle-income countries in Africa. The large endemicity and escalating threat of diseases such as malaria and arboviral diseases, intensified by climate change, disproportionately affects vulnerable communities globally. We highlight the urgency of prioritizing research and development, advocating for robust scientific inquiry to promote adaptation strategies, and the vital role that the next generation of African research leaders will play in addressing these challenges. Despite significant challenges such as funding shortages within countries, various pan-African-oriented funding bodies such as the African Academy of Sciences, the Africa Research Excellence Fund, the Wellcome Trust, the U.S. National Institutes of Health, and the Bill and Melinda Gates Foundation as well as initiatives such as the African Research Initiative for Scientific Excellence and the Pan-African Mosquito Control Association, have empowered (or are empowering) these researchers by supporting capacity building activities, including continental and global networking, skill development, mentoring, and African-led research. This article underscores the urgency of increased national investment in research, proposing the establishment of research government agencies to drive evidence-based interventions. Collaboration between governments and scientific communities, sustained by pan-African funding bodies, is crucial. Through these efforts, African nations are likely to enhance the resilience and adaptive capacity of their systems and communities by navigating these challenges effectively, fostering scientific excellence and implementing transformative solutions against climate-sensitive vector-borne diseases.

Physiological correlates of ecological divergence along an urbanization gradient: differential tolerance to ammonia among molecular forms of the malaria mosquito Anopheles gambiae.

BACKGROUND: Limitations in the ability of organisms to tolerate environmental stressors affect their fundamental ecological niche and constrain their distribution to specific habitats. Evolution of tolerance, therefore, can engender ecological niche dynamics. Forest populations of the afro-tropical malaria mosquito Anopheles gambiae have been shown to adapt to historically unsuitable larval habitats polluted with decaying organic matter that are found in densely populated urban agglomerates of Cameroon. This process has resulted in niche expansion from rural to urban environments that is associated with cryptic speciation and ecological divergence of two evolutionarily significant units within this taxon, the molecular forms M and S, among which reproductive isolation is significant but still incomplete. Habitat segregation between the two forms results in a mosaic distribution of clinally parapatric patches, with the M form predominating in the centre of urban agglomerates and the S form in the surrounding rural localities. We hypothesized that development of tolerance to nitrogenous pollutants derived from the decomposition of organic matter, among which ammonia is the most toxic to aquatic organisms, may affect this pattern of distribution and process of niche expansion by the M form. RESULTS: Acute toxicity bioassays indicated that populations of the two molecular forms occurring at the extremes of an urbanization gradient in Yaounde, the capital of Cameroon, differed in their response to ammonia. The regression lines best describing the dose-mortality profile differed in the scale of the explanatory variable (ammonia concentration log-transformed for the S form and linear for the M form), and in slope (steeper for the S form and shallower for the M form). These features reflected differences in the frequency distribution of individual tolerance thresholds in the two populations as assessed by probit analysis, with the M form exhibiting a greater mean and variance compared to the S form. CONCLUSIONS: In agreement with expectations based on the pattern of habitat partitioning and exposure to ammonia in larval habitats in Yaounde, the M form showed greater tolerance to ammonia compared to the S form. This trait may be part of the physiological machinery allowing forest populations of the M form to colonize polluted larval habitats, which is at the heart of its niche expansion in densely populated human settlements in Cameroon.

Towards the invasion of wild and rural forested areas in Gabon (Central Africa) by the Asian tiger mosquito Aedes albopictus: Potential risks from the one health perspective.

BACKGROUND: Since its first record in urban areas of Central-Africa in the 2000s, the invasive mosquito, Aedes albopictus, has spread throughout the region, including in remote villages in forested areas, causing outbreaks of Aedes-borne diseases, such as dengue and chikungunya. Such invasion might enhance Ae. albopictus interactions with wild animals in forest ecosystems and favor the spillover of zoonotic arboviruses to humans. The aim of this study was to monitor Ae. albopictus spread in the wildlife reserve of La Lopé National Park (Gabon), and evaluate the magnitude of the rainforest ecosystem colonization. METHODOLOGY: From 2014 to 2018, we used ovitraps, larval surveys, BG-Sentinel traps, and human landing catches along an anthropization gradient from La Lopé village to the natural forest in the Park. CONCLUSIONS: We detected Ae. albopictus in gallery forest up to 15 km away from La Lopé village. However, Ae. albopictus was significantly more abundant at anthropogenic sites than in less anthropized areas. The number of eggs laid by Ae. albopictus decreased progressively with the distance from the forest fringe up to 200m inside the forest. Our results suggested that in forest ecosystems, high Ae. albopictus density is mainly observed at interfaces between anthropized and natural forested environments. Additionally, our data suggested that Ae. albopictus may act as a bridge vector of zoonotic pathogens between wild and anthropogenic compartments.

Urban Green Spaces and Vector-Borne Disease Risk in Africa: The Case of an Unclean Forested Park in Libreville (Gabon, Central Africa).

In Africa, vector-borne diseases are a major public health issue, especially in cities. Urban greening is increasingly considered to promote inhabitants' well-being. However, the impact of urban green spaces on vector risk remains poorly investigated, particularly urban forests in poor hygienic conditions. Therefore, using larval sampling and human landing catches, this study investigated the mosquito diversity and the vector risk in a forest patch and its inhabited surroundings in Libreville, Gabon, central Africa. Among the 104 water containers explored, 94 (90.4%) were artificial (gutters, used tires, plastic bottles) and 10 (9.6%) were natural (puddles, streams, tree holes). In total, 770 mosquitoes belonging to 14 species were collected from such water containers (73.1% outside the forested area). The mosquito community was dominated by Aedes albopictus (33.5%), Culex quinquefasciatus (30.4%), and Lutzia tigripes (16.5%). Although mosquito diversity was almost double outside compared to inside the forest (Shannon diversity index: 1.3 vs. 0.7, respectively), the species relative abundance (Morisita-Horn index = 0.7) was similar. Ae. albopictus (86.1%) was the most aggressive species, putting people at risk of Aedes-borne viruses. This study highlights the importance of waste pollution in urban forested ecosystems as a potential driver of mosquito-borne diseases.

Genetic isolation within the malaria mosquito Anopheles melas.

Anopheles melas is a brackish water-breeding member of the Anopheles gambiae complex that is distributed along the coast of West Africa and is a major malaria vector within its range. Because little is known about the population structure of this species, we analysed 15 microsatellite markers and 1161 bp of mtDNA in 11 A. melas populations collected throughout its range. Compared with its sibling species A. gambiae, A. melas populations have a high level of genetic differentiation between them, representing its patchy distribution due to its fragmented larval habitat that is associated with mangroves and salt marsh grass. Populations clustered into three distinct groups representing Western Africa, Southern Africa and Bioko Island populations that appear to be mostly isolated. Fixed differences in the mtDNA are present between all three clusters, and a Bayesian clustering analysis of the microsatellite data found no evidence for migration from mainland to Bioko Island populations, and little migration was evident between the Southern to the Western cluster. Surprisingly, mtDNA divergence between the three A. melas clusters is on par with levels of divergence between other species of the A. gambiae complex, and no support for monophyly was observed in a maximum-likelihood phylogenetic analysis. Finally, an approximate Bayesian analysis of microsatellite data indicates that Bioko Island A. melas populations were connected to the mainland populations in the past, but became isolated, presumably when sea levels rose after the last glaciation period (≥10 000-11 000 bp). This study has exposed species-level genetic divergence within A. melas and also has implications for control of this malaria vector.

High mosquito burden and malaria transmission in a district of the city of Douala, Cameroon.

BACKGROUND: Rapid demographic growth in Douala city, Cameroon, has resulted in profound ecological and environmental changes. Although demographic changes can affect anopheline mosquito breeding sites, there is a lack of understanding about the epidemiological impact that such changes might have on vector ecology and malaria transmission. METHODS: A 12-month entomological study was conducted in a highly populated district of Douala called Ndogpassi. Adult mosquitoes were collected using two methods: 1) human landing catches (HLC); and 2) Centers for Disease Control and Prevention (CDC) light traps; these methods were used twice monthly from January to December 2011. Mosquito genus and species were identified with morphological and molecular diagnostic tools. The sampling efficiency of the CDC light trap and HLC were compared. Anopheles gambiae infection with Plasmodium falciparum was detected using ELISA. Susceptibility to DDT, permethrin, and deltamethrin insecticides were also determined. RESULTS: A total of 6923 mosquitoes were collected by HLC (5198) and CDC light traps (1725). There was no equivalence in the sampling efficiency between light traps and human landing catches (P > 0.01). With 51% of the total, Culex was the most common, followed by Anopheles (26.14%), Mansonia (22.7%) and Aedes (0.1%). An. gambiae ss (M form) comprised ~98% of the total anophelines collected. An. gambiae had a biting rate of 0.25 to 49.25 bites per human per night, and was the only species found to be infected with P. falciparum. A P. falciparum infection rate of 0.5% was calculated (based on enzyme-linked immunosorbent assays using the circumsporozoite surface protein). The entomological inoculation rate was estimated at 31 infective bites per annum. Insecticide susceptibility tests on An. gambiae females revealed a mortality rate of 33%, 76% and 98% for DDT, permethrin and deltamethrin, respectively. The West African kdr allele (L1014F) was detected in 38 of the 61 An. gambiae analyzed (62.3%). CONCLUSIONS: The present study revealed seasonal malaria transmission in Douala. High levels of An. gambiae were detected along with a high prevalence of insecticide resistance in this vector population. These findings highlight the need to promote use of insecticide-impregnated bed nets in Douala.

New cytotoxic obacunone-type limonoid and others constituents from the stem bark of Carapa procera DC (Meliaceae).

The phytochemical study of the CH2Cl2- MeOH (1:1, v/v) extract of the stem bark of Carapa procera DC (Meliaceae) led to the isolation and characterisation of a new natural limonoid 7ß-obacunol (6), along with seven known compounds. Their structures were elucidated by spectroscopic means, including 1 D and 2 D NMR, HRESI-MS and by comparison with published data. The cytotoxicity of compounds 1-6 was assessed in vitro by the WST-1 assay on human lung adenocarcinoma A549 and Raw 264.7 mouse macrophage cell lines. Results suggested that obacunone (3) exhibited the most potent cytotoxic effect against A549 and Raw 264.7 cells with respective IC50 values of 25.24 µM and 29.14 µM, while the new natural limonoid 7ß-obacunol (6) exhibited 32.75 µM and 39.19 µM, respectively on both cell lines. Therefore, limonoid derivatives might be promising sources of natural bioactive metabolites against cancer.

Systematic Review on Diversity and Distribution of Anopheles Species in Gabon: A Fresh Look at the Potential Malaria Vectors and Perspectives.

Gabon is located in the malaria hyper-endemic zone, where data concerning malaria vector distribution remains fragmentary, making it difficult to implement an effective vector control strategy. Thus, it becomes crucial and urgent to undertake entomological surveys that will allow a better mapping of the Anopheles species present in Gabon. In this review, we examined different articles dealing with Anopheles in Gabon from ProQuest, Web of Science, PubMed, and Google scholar databases. After applying the eligibility criteria to 7543 articles collected from four databases, 42 studies were included that covered a 91-year period of study. The review revealed a wide diversity of Anopheles species in Gabon with a heterogeneous distribution. Indeed, our review revealed the presence of 41 Anopheles species, of which the most abundant were members of the Gambiae and Nili complexes and those of the Funestus and Moucheti groups. However, our review also revealed that the major and minor vectors of malaria in Gabon are present in both sylvatic, rural, and urban environments. The observation of human malaria vectors in sylvatic environments raises the question of the role that the sylvatic environment may play in maintaining malaria transmission in rural and urban areas. Ultimately, it appears that knowledge of biodiversity and spatial distribution of Anopheles mosquitoes is fragmentary in Gabon, suggesting that additional studies are necessary to complete and update these entomological data, which are useful for the implementation of vector control strategies.

The genome sequence of the malaria mosquito, Anopheles funestus, Giles, 1900.

We present a genome assembly from an individual female Anopheles funestus (the malaria mosquito; Arthropoda; Insecta; Diptera; Culicidae). The genome sequence is 251 megabases in span. The majority of the assembly is scaffolded into three chromosomal pseudomolecules with the X sex chromosome assembled. The complete mitochondrial genome was also assembled and is 15.4 kilobases in length.

Experimental infections with Zika virus strains reveal high vector competence of Aedes albopictus and Aedes aegypti populations from Gabon (Central Africa) for the African virus lineage.

ABSTRACTThe two main Zika virus (ZIKV) vectors, Aedes albopictus and Aedes aegypti (invasive and native species, respectively), are present in Gabon (Central Africa). The aim of this study was to determine the entomological ZIKV risk associated with these mosquito species in Gabon by evaluating their vector competence for an African (i.e. representative of the endemic strains circulating in sub-Saharan Africa) and two Asian (i.e. representatives of exogenous epidemic strains that could be introduced) ZIKV strains. The transmission efficiency of one Ae. aegypti and two Ae. albopictus field-collected populations from Libreville and Franceville was assayed at day 7, 14 and 21 after experimental oral infection. The two mosquito species could transmit all three ZIKV strains already at day 7 post-infection, but transmission efficiency was higher for the African strain than the non-African strains (>60% versus <14%; incubation period of 14-21 days). The two mosquito species exhibited comparable vector competence for ZIKV, although the amount of viral particles (African strain) in saliva was significantly higher in Ae. albopictus than Ae. aegypti at day 14 post-infection. These findings suggest that overall, ZIKV risk in Gabon is mainly related to virus strains that circulate endemically across sub-Saharan Africa, although the transmission of non-African strains remain possible in case of introduction. Due to its high infestation indexes and ecological/geographical ranges, this risk appears mainly associated with Ae. albopictus. Vector surveillance and control methods against this invasive mosquito must be strengthened in the region to limit the risk of future outbreaks.

Ecological plasticity to ions concentration determines genetic response and dominance of Anopheles coluzzii larvae in urban coastal habitats of Central Africa.

In Central Africa, the malaria vector Anopheles coluzzii is predominant in urban and coastal habitats. However, little is known about the environmental factors that may be involved in this process. Here, we performed an analysis of 28 physicochemical characteristics of 59 breeding sites across 5 urban and rural sites in coastal areas of Central Africa. We then modelled the relative frequency of An. coluzzii larvae to these physicochemical parameters in order to investigate environmental patterns. Then, we assessed the expression variation of 10 candidate genes in An. coluzzii, previously incriminated with insecticide resistance and osmoregulation in urban settings. Our results confirmed the ecological plasticity of An. coluzzii larvae to breed in a large range of aquatic conditions and its predominance in breeding sites rich in ions. Gene expression patterns were comparable between urban and rural habitats, suggesting a broad response to ions concentrations of whatever origin. Altogether, An. coluzzii exhibits a plastic response to occupy both coastal and urban habitats. This entails important consequences for malaria control in the context of the rapid urban expansion in Africa in the coming years.

Population genetic structure of the malaria vector Anopheles nili in sub-Saharan Africa.

BACKGROUND: Anopheles nili is a widespread efficient vector of human malaria parasites in the humid savannas and forested areas of sub-Saharan Africa. Understanding An. nili population structure and gene flow patterns could be useful for the development of locally-adapted vector control measures. METHODS: Polymorphism at eleven recently developed microsatelitte markers, and sequence variation in four genes within the 28s rDNA subunit (ITS2 and D3) and mtDNA (COII and ND4) were assessed to explore the level of genetic variability and differentiation among nine populations of An. nili from Senegal, Ivory Coast, Burkina Faso, Nigeria, Cameroon and the Democratic Republic of Congo (DRC). RESULTS: All microsatellite loci successfully amplified in all populations, showing high and very similar levels of genetic diversity in populations from West Africa and Cameroon (mean Rs = 8.10-8.88, mean He = 0.805-0.849) and much lower diversity in the Kenge population from DRC (mean Rs = 5.43, mean He = 0.594). Bayesian clustering analysis of microsatellite allelic frequencies revealed two main genetic clusters in the dataset. The first one included only the Kenge population and the second grouped together all other populations. High Fst estimates based on microsatellites (Fst > 0.118, P < 0.001) were observed in all comparisons between Kenge and all other populations. By contrast, low Fst estimates (Fst < 0.022, P < 0.05) were observed between populations within the second cluster. The correlation between genetic and geographic distances was weak and possibly obscured by demographic instability. Sequence variation in mtDNA genes matched these results, whereas low polymorphism in rDNA genes prevented detection of any population substructure at this geographical scale. CONCLUSION: Overall, high genetic homogeneity of the An. nili gene pool was found across its distribution range in West and Central Africa, although demographic events probably resulted in a higher level of genetic isolation in the marginal population of Kenge (DRC). The role of the equatorial forest block as a barrier to gene flow and the implication of such findings for vector control are discussed.

First report of Anopheles (Cellia) multicolor during a study of tolerance to salinity of Anopheles arabiensis larvae in Nouakchott, Mauritania.

BACKGROUND: Anopheles multicolor is known to be present in the arid areas of Africa north of the Sahara Desert, especially in oases. To date, its presence in Mauritania has not been reported. Here, we present the first record of its presence in Nouakchott, the capital of Mauritania. The larvae of An. multicolor, together with those of An. arabiensis, the major malaria vector in the city, were found thriving in highly saline surface water collections. METHODS: Entomological surveys were carried out during 2016-2017 in Nouakchott. Mosquito larval habitats were investigated through larval surveys while indoor resting culicid fauna were collected using hand-held aspirator. Physicochemical parameters of the larval habitats were measured on-site, at the time mosquitoes were collected. Larvae and pupae were reared to adults in the insectaries. Morphological and polymerase chain reaction (PCR)-based methods were used to identify newly emerged adults. Batches of fourth-instar larvae were used to assess salinity tolerance by exposing them to increasing concentrations of NaCl, and mortality was monitored throughout development. RESULTS: Morphological and molecular results confirmed that the specimens were An. multicolor and An. arabiensis. Sequences of 24 An. multicolor adult mosquitoes showed 100% nucleotide identity with the published sequences of An. multicolor from Iran. The physicochemical analysis of the water from the two larval habitats revealed highly saline conditions, with NaCl content ranging between 16.8 and 28.9 g/l (i.e. between c.50-80% seawater). Anopheles multicolor and An. arabiensis fourth-instar larvae survival rates at 17.5 g/l NaCl were 86.5% and 75%, respectively. Anopheles arabiensis larvae showed variable levels of salt tolerance according to the larval habitat. Adult An. multicolor specimens were collected resting indoor at low frequency (0.7%) compared to the other culicid mosquitoes. CONCLUSIONS: To the best of our knowledge, this paper is the first report of An. multicolor in Mauritania, extending the known distributional range of the species to the south, as well as to the west. Highly salt-tolerant populations of An. arabiensis and An. multicolor were observed. Because salt-water collections are widespread in Nouakchott, the relevance of these findings for the dynamics and epidemiology of malaria transmission needs to be assessed.

Molecular genetic investigation of morphological species of members of the Neomelaniconion subgenus (Diptera: Culicidae: Aedini) from Madagascar using ribosomal internal transcribed spacer 2.

Species of Neomelaniconion (Neo.) are aggressive toward humans and domestic animals. Some of these species play a role in the transmission of arbovirus. In Madagascar, separation of these arbovirus vectors was based on doubtful morphological features. Sequence variation of the ribosomal DNA internal transcribed spacer 2 from morphological Neomelaniconion species of the Neomelaniconion subgenus from Madagascar was investigated to assess the validity of morphologically defined taxa. Both methods showed two taxonomic groups: the Circumluteolus Group that includes Aedes (Neo.) belleci (Le Goff, Bousses, and Brunhes, 2007), Ae. (Neo.) nigropterum (Le Goff, Bousses, and Brunhes, 2007), and Ae. (Neo.) circumluteolus (Theobald, 1908); and the Sylvaticum Group that includes Ae. (Neo.) sylvaticum (Le Goff, Bousses,and Brunhes, 2007), Ae. (Neo.) fontenillei (Le Goff, Bousses, and Brunhes, 2007), and Ae. (Neo.) albiradius (Le Goff, Bousses, and Brunhes, 2007). Although sequence analysis failed to completely resolve each group, four distinct patterns of sequence variation supported by several fixed mutations were shown from specimens in Madagascar.

Natural Wolbachia infections are common in the major malaria vectors in Central Africa.

During the last decade, the endosymbiont bacterium Wolbachia has emerged as a biological tool for vector disease control. However, for long time, it was believed that Wolbachia was absent in natural populations of Anopheles. The recent discovery that species within the Anopheles gambiae complex host Wolbachia in natural conditions has opened new opportunities for malaria control research in Africa. Here, we investigated the prevalence and diversity of Wolbachia infection in 25 African Anopheles species in Gabon (Central Africa). Our results revealed the presence of Wolbachia in 16 of these species, including the major malaria vectors in this area. The infection prevalence varied greatly among species, confirming that sample size is a key factor to detect the infection. Moreover, our sequencing and phylogenetic analyses showed the important diversity of Wolbachia strains that infect Anopheles. Co-evolutionary analysis unveiled patterns of Wolbachia transmission within some Anopheles species, suggesting that past independent acquisition events were followed by co-cladogenesis. The large diversity of Wolbachia strains that infect natural populations of Anopheles offers a promising opportunity to select suitable phenotypes for suppressing Plasmodium transmission and/or manipulating Anopheles reproduction, which in turn could be used to reduce the malaria burden in Africa.

A new species in the major malaria vector complex sheds light on reticulated species evolution.

Complexes of closely related species provide key insights into the rapid and independent evolution of adaptive traits. Here, we described and studied Anopheles fontenillei sp.n., a new species in the Anopheles gambiae complex that we recently discovered in the forested areas of Gabon, Central Africa. Our analysis placed the new taxon in the phylogenetic tree of the An. gambiae complex, revealing important introgression events with other members of the complex. Particularly, we detected recent introgression, with Anopheles gambiae and Anopheles coluzzii, of genes directly involved in vectorial capacity. Moreover, genome analysis of the new species allowed us to clarify the evolutionary history of the 3La inversion. Overall, An. fontenillei sp.n. analysis improved our understanding of the relationship between species within the An. gambiae complex, and provided insight into the evolution of vectorial capacity traits that are relevant for the successful control of malaria in Africa.

Identification and geographic distribution of the ACE-1R mutation in the malaria vector Anopheles gambiae in south-western Burkina Faso, West Africa.

Resistance of Anopheles gambiae to organophosphate and carbamate insecticides was first reported in Côte d'Ivoire, West Africa. Subsequent studies revealed that it resulted from a single point mutation in the oxyanion hole of the acetycholinesterase enzyme (ace-1(R) mutation). We investigated the distribution and prevalence of the ace-1(R) mutation in An. gambiae s.l. populations from seven locations in south-western Burkina Faso. The ace-1(R) mutation was found in both M and S molecular forms of An. gambiae s.s., but it was absent in An. arabiensis. Its frequency ranged from 0.25 to 0.5 in S form and 0.04 to 0.13 in M form, though they were sympatric. The lack of homozygous resistance indicated a strong genetic cost associated with the mutation. These data suggest that organophosphate and carbamate resistance conferred by target site insensitivity is spreading in populations of An. gambiae s.s. from West Africa.

Population structure of the malaria vector Anopheles moucheti in the equatorial forest region of Africa.

BACKGROUND: Anopheles moucheti is a major malaria vector in forested areas of Africa. However, despite its important epidemiological role, it remains poorly known and insufficiently studied. Here, levels of genetic differentiation were estimated between different A. moucheti populations sampled throughout its distribution range in Central Africa. METHODS: Polymorphism at ten microsatellite markers was compared in mosquitoes sampled in Cameroon, the Democratic Republic of Congo and an island on Lake Victoria in Uganda. Microsatellite data were used to estimate genetic diversity within populations, their relative long-term effective population size, and the level of genetic differentiation between them. RESULTS: All specimens collected in Tsakalakuku (Democratic Republic of Congo) were identified as A. m. bervoetsi while other samples consisted of A. m. moucheti. Successful amplification was obtained at all microsatellite loci within all A. m. moucheti samples while only six loci amplified in A. m. bervoetsi. Allelic richness and heterozygosity were high for all populations except the island population of Uganda and A. m. bervoetsi. High levels of genetic differentiation were recorded between A. m. bervoetsi and each A. m. moucheti sample as well as between the island population of A. m. moucheti and mainland populations. Significant isolation by distance was evidenced between mainland populations. CONCLUSION: High levels of genetic differentiation supports complete speciation of A. m. bervoetsi which should henceforth be recognized as a full species and named A. bervoetsi. Isolation by distance is the main force driving differentiation between mainland populations of A. m. moucheti. Genetically and geographically isolated populations exist on Lake Victoria islands, which might serve as relevant field sites for evaluation of innovative vector control strategies.