Extensive variation and strain-specificity in dengue virus susceptibility among African Aedes aegypti populations.

African populations of the mosquito Aedes aegypti are usually considered less susceptible to infection by human-pathogenic flaviviruses than globally invasive populations found outside Africa. Although this contrast has been well documented for Zika virus (ZIKV), it is unclear to what extent it is true for dengue virus (DENV), the most prevalent flavivirus of humans. Addressing this question is complicated by substantial genetic diversity among DENV strains, most notably in the form of four genetic types (DENV1 to DENV4), that can lead to genetically specific interactions with mosquito populations. Here, we carried out a survey of DENV susceptibility using a panel of seven field-derived Ae. aegypti colonies from across the African range of the species and a colony from Guadeloupe, French West Indies as non-African reference. We found considerable variation in the ability of African Ae. aegypti populations to acquire and replicate a panel of six DENV strains spanning the four DENV types. Although African Ae. aegypti populations were generally less susceptible than the reference non-African population from Guadeloupe, in several instances some African populations were equally or more susceptible than the Guadeloupe population. Moreover, the relative level of susceptibility between African mosquito populations depended on the DENV strain, indicating genetically specific interactions. We conclude that unlike ZIKV susceptibility, there is no clear-cut dichotomy in DENV susceptibility between African and non-African Ae. aegypti. DENV susceptibility of African Ae. aegypti populations is highly heterogeneous and largely governed by the specific pairing of mosquito population and DENV strain.

Distribution and insecticide resistance profile of the major malaria vector Anopheles funestus group across the African continent.

There has been significant progress in malaria control in the last 2 decades, with a decline in mortality and morbidity. However, these gains are jeopardised by insecticide resistance, which negatively impacts the core interventions, such as insecticide-treated nets (ITN) and indoor residual spraying (IRS). While most malaria control and research efforts are still focused on Anopheles gambiae complex mosquitoes, Anopheles funestus remains an important vector in many countries and, in some cases, contributes to most of the local transmission. As countries move towards malaria elimination, it is important to ensure that all dominant vector species, including An. funestus, an important vector in some countries, are targeted. The objective of this review is to compile and discuss information related to A. funestus populations' resistance to insecticides and the mechanisms involved across Africa, emphasising the sibling species and their resistance profiles in relation to malaria elimination goals. Data on insecticide resistance in An. funestus malaria vectors in Africa were extracted from published studies. Online bibliographic databases, including Google Scholar and PubMed, were used to search for relevant studies. Articles published between 2000 and May 2023 reporting resistance of An. funestus to insecticides and associated mechanisms were included. Those reporting only bionomics were excluded. Spatial variation in species distribution and resistance to insecticides was recorded from 174 articles that met the selection criteria. It was found that An. funestus was increasingly resistant to the four classes of insecticides recommended by the World Health Organisation for malaria vector control; however, this varied by country. Insecticide resistance appears to reduce the effectiveness of vector control methods, particularly IRS and ITN. Biochemical resistance due to detoxification enzymes (P450s and glutathione-S-transferases [GSTs]) in An. funestus was widely recorded. However, An. funestus in Africa remains susceptible to other insecticide classes, such as organophosphates and neonicotinoids. This review highlights the increasing insecticide resistance of An. funestus mosquitoes, which are important malaria vectors in Africa, posing a significant challenge to malaria control efforts. While An. funestus has shown resistance to the recommended insecticide classes, notably pyrethroids and, in some cases, organochlorides and carbamates, it remains susceptible to other classes of insecticides such as organophosphates and neonicotinoids, providing potential alternative options for vector control strategies. The study underscores the need for targeted interventions that consider the population structure and geographical distribution of An. funestus, including its sibling species and their insecticide resistance profiles, to effectively achieve malaria elimination goals.

Des progrès importants ont été réalisés dans le contrôle du paludisme au cours des deux dernières décennies, qui se traduisent par une baisse de la mortalité et de la morbidité. Cependant, ces gains sont compromis par la résistance aux insecticides, ce qui a un impact négatif sur les interventions de base, telles que les moustiquaires imprégnées d'insecticides et la pulvérisation intradomicilliare (PID). Alors que la plupart des efforts de contrôle et de recherche sur le paludisme sont toujours axés sur les moustiques du complexes Anopheles gambiae, Anopheles funestus reste un vecteur important dans de nombreux pays et, dans certains cas, contribue à la majeure partie de la transmission locale. Au moment où certains pays se dirigent vers l'élimination du paludisme, il serait important de prendre en considération toutes les espèces vectrices dominantes, y compris An. funestus. L'objectif de cette revue est de compiler et de discuter des informations liées à la résistance des populations d'An. funestus aux insecticides et les mécanismes impliqués à travers l'Afrique, en mettant l'accent sur les sous espèces et leurs profils de résistance en relation avec les objectifs d'élimination du paludisme. Les données sur la résistance aux insecticides chez An. funestus vecteurs du paludisme en Afrique ont été extraites d'études publiées dans des bases de données bibliographiques comme Google Scholar et PubMed. Les articles publiés entre 2000 et mai 2023, rapportant la résistance de An. funestus aux insecticides et les mécanismes associés ont été inclus. Ceux portant uniquement sur la bionomie ont été exclus. Au total 174 articles portant sur la variation spatiale de la résistance des espèces du groupe An. funestus aux insecticides répondaient aux critères de sélection. De ces analyses, il ressort qu'An. funestus était de plus en plus résistant aux quatre classes d'insecticides recommandées par l'Organisation Mondiale de la Santé (OMS) pour le contrôle des vecteurs du paludisme ce qui semble réduire l'efficacité des méthodes de contrôle des vecteurs, en particulier les moustiquaires imprégnées d'insecticide et la pulvérisation intradomiciliaire. avec des variations en fonction des pays. Les mécanismes de résistance aux insecticides de type biochimique liée aux enzymes de détoxification (P450S et GST) ont été largement rapportés chez An. funestus. De nombreux gènes P450 associés à la résistance métabolique ont été mis en évidence chez An. funestus collecté sur le terrain. Cependant, An. funestus en Afrique reste sensible à d'autres classes d'insecticides, telles que les organophosphorés et les néonicotinoïdes. La résistance aux insecticides. Cette revue met en évidence la résistance croissante aux insecticides chez les moustiques du groupe Funestus, un vecteur important du paludisme en Afrique, posant ainsi un défi important aux efforts de contrôle du paludisme. Tandis que An. funestus a montré une résistance aux classes d'insecticide recommandées, notamment les pyréthroïdes et, dans certains cas, les organochlorés et les carbamates, il reste sensible à d'autres classes d'insecticides tels que les organophosphorés et les néonicotinoïdes, offrant des options alternatives potentielles de contrôle des vecteurs. L'étude souligne la nécessité d'interventions ciblées qui considèrent la structure de la population et la distribution géographique d'An. funestus, y compris ses sous espèces et leurs profils de résistance aux insecticides, pour atteindre efficacement les objectifs d'élimination du paludisme.

Adhesion and virulence properties of native Metarhizium fungal strains from Burkina Faso for the control of malaria vectors.

BACKGROUND: Local strains of the entomopathogenic fungus Metarhizium pingshaense in Burkina Faso have demonstrated remarkable virulence against malaria vectors, positioning them as promising candidates for inclusion in the future arsenal of malaria control strategies. However, the underlying mechanisms responsible for this virulence remain unknown. To comprehend the fungal infection process, it is crucial to investigate the attachment mechanisms of fungal spores to the mosquito cuticle and explore the relationship between virulence and attachment kinetics. This study aims to assess the adhesion and virulence properties of native Metarhizium fungal strains from Burkina Faso for controlling malaria vectors. METHODS: Fungal strains were isolated from 201 insects and 1399 rhizosphere samples, and four strains of Metarhizium fungi were selected. Fungal suspensions were used to infect 3-day-old female Anopheles coluzzii mosquitoes at three different concentrations (106, 107, 108 conidia/ml). The survival of the mosquitoes was measured over 14 days, and fungal growth was quantified after 1 and 24 h to assess adhesion of the fungal strains onto the mosquito cuticle. RESULTS: All four fungi strains increased mosquito mortality compared to control (Chi-square test, χ2 = 286.55, df = 4, P < 0.001). Adhesion of the fungal strains was observed on the mosquito cuticle after 24 h at high concentrations (1 × 108 conidia/ml), with one strain, having the highest virulent, showing adhesion after just 1 h. CONCLUSION: The native strains of Metarhizium spp. fungi found in Burkina Faso have the potential to be effective biocontrol agents against malaria vectors, with some strains showing high levels of both virulence and adhesion to the mosquito cuticle.

Advances in the genetic characterization of the malaria vector, Anopheles funestus, and implications for improved surveillance and control.

Anopheles mosquitoes present a major public health challenge in sub-Saharan Africa; notably, as vectors of malaria that kill over half a million people annually. In parts of the east and southern Africa region, one species in the Funestus group, Anopheles funestus, has established itself as an exceptionally dominant vector in some areas, it is responsible for more than 90% of all malaria transmission events. However, compared to other malaria vectors, the species is far less studied, partly due to difficulties in laboratory colonization and the unresolved aspects of its taxonomy and systematics. Control of An. funestus is also increasingly difficult because it has developed widespread resistance to public health insecticides. Fortunately, recent advances in molecular techniques are enabling greater insights into species identity, gene flow patterns, population structure, and the spread of resistance in mosquitoes. These advances and their potential applications are reviewed with a focus on four research themes relevant to the biology and control of An. funestus in Africa, namely: (i) the taxonomic characterization of different vector species within the Funestus group and their role in malaria transmission; (ii) insecticide resistance profile; (iii) population genetic diversity and gene flow, and (iv) applications of genetic technologies for surveillance and control. The research gaps and opportunities identified in this review will provide a basis for improving the surveillance and control of An. funestus and malaria transmission in Africa.

Methanol and acetone extracts from the leaves of selected aromatic plants affect survival of field collected Anopheles arabiensis (Diptera: Culicidae) from Kisumu, Kenya.

The massive and inappropriate use of synthetic insecticides is causing significant and increasing environmental disruption. Therefore, developing effective natural mosquitocidal compounds could be an alternative tool for malarial vector control. The present study investigates the larvicidal and adulticidal effect of methanol and acetone extracts of leaves from Lippia chevalieri, Lippia multiflora, Cymbopogon schoenanthus, and Lantana camara against Anopheles arabiensis, to control the most widespread vector transmitting malaria in sub-Saharan. Africa. Extracts were evaluated following WHO modified test procedure against third- to fourth-instar larvae and, non-blood-fed females from 3- to 5-day-old field populations of An. arabiensis under laboratory conditions using WHO larval and CDC bottle bioassays, respectively. Mortality was recorded after 24-h exposure and several compounds were identified in the extracts. The methanolic and acetonic extracts of L. camara were effective against larvae showing lethal concentrations to 50% (LC50) of the population, at 89.48 and 58.72 ppm, respectively. The acetonic extracts of C. schoenanthus and L. chevalieri showed higher toxicities LC50s of 0.16% and 0.22% against female adults, respectively. The methanolic extracts of L. multiflora and L. chevalieri LC50s were effective at 0.17% and 0.27%, respectively, against female adults. These results indicate that the plant extracts tested may represent effective means to control An. arabiensis when used to treat the surface of the marshes.

Association of 410L, 1016I and 1534C kdr mutations with pyrethroid resistance in Aedes aegypti from Ouagadougou, Burkina Faso, and development of a one-step multiplex PCR method for the simultaneous detection of 1534C and 1016I kdr mutations.

BACKGROUND: Since 2000, Burkina Faso has experienced regular dengue cases and outbreaks, making dengue an increasingly important health concern for the country. Previous studies in Burkina Faso reported that resistance of Aedes aegypti to pyrethroid insecticides was associated with the F1534C and V1016I kdr mutations. The current study reports high resistance of Ae. aegypti populations to pyrethroid insecticides, likely supported by mutations in the voltage-gated sodium channel, here evidenced by genotyping the kdr SNPs V410L, V1016I and F1534C. We also describe a new multiplex PCR-based diagnostic of F1534C and V1016I kdr SNPs. METHODS: Larvae of Ae. aegypti were collected from three health districts of Ouagadougou in 2018. The resistance status of Ae. aegypti to permethrin (15 µg/ml) and deltamethrin (10 µg/ml) was tested using bottles and to malathion (5%) using WHO tube tests. All bioassays used 1-h exposure and mortality recorded 24 h post-exposure. Bioassay results were interpreted according to WHO thresholds for resistance diagnosis. The kdr mutations were screened using AS-PCR and TaqMan methods in exposed and non-exposed Aedes mosquitoes. RESULTS: Females from all health districts were resistant to permethrin and deltamethrin (< 20% mortality) but were fully susceptible to 5% malathion. The F1534C and V1016I kdr mutations were successfully detected using a newly developed multiplex PCR in perfect agreement with TaqMan method. The 1534C/1016I/410L haplotype was correlated with permethrin resistance but not with deltamethrin resistance; however, the test power was limited by a low frequency of dead individuals in deltamethrin exposure. CONCLUSIONS: Resistance to pyrethroid insecticides is associated with kdr mutant haplotypes, while the absence of substantial resistance to malathion suggests that it remains a viable option for dengue vector control in Ouagadougou.

Dating the origin and spread of specialization on human hosts in Aedes aegypti mosquitoes.

The globally invasive mosquito subspecies Aedes aegypti aegypti is an effective vector of human arboviruses, in part because it specializes in biting humans and breeding in human habitats. Recent work suggests that specialization first arose as an adaptation to long, hot dry seasons in the West African Sahel, where Ae. aegypti relies on human-stored water for breeding. Here, we use whole-genome cross-coalescent analysis to date the emergence of human-specialist populationsand thus further probe the climate hypothesis. Importantly, we take advantage of the known migration of specialists out of Africa during the Atlantic Slave Trade to calibrate the coalescent clock and thus obtain a more precise estimate of the older evolutionary event than would otherwise be possible. We find that human-specialist mosquitoes diverged rapidly from ecological generalists approximately 5000 years ago, at the end of the African Humid Period-a time when the Sahara dried and water stored by humans became a uniquely stable, aquatic niche in the Sahel. We also use population genomic analyses to date a previously observed influx of human-specialist alleles into major West African cities. The characteristic length of tracts of human-specialist ancestry present on a generalist genetic background in Kumasi and Ouagadougou suggests the change in behavior occurred during rapid urbanization over the last 20-40 years. Taken together, we show that the timing and ecological context of two previously observed shifts towards human biting in Ae. aegypti differ; climate was likely the original driver, but urbanization has become increasingly important in recent decades.

Biology and Behaviour of Aedes aegypti in the Human Environment: Opportunities for Vector Control of Arbovirus Transmission.

Aedes aegypti is a ubiquitous vector of arboviruses mostly in urbanised areas throughout the tropics and subtropics and a growing threat beyond. Control of Ae. aegypti is difficult and costly, and no vaccines are available for most of the viruses it transmits. With practical control solutions our goal, ideally suitable for delivery by householders in affected communities, we reviewed the literature on adult Ae. aegypti biology and behaviour, within and close to the human home, the arena where such interventions must impact. We found that knowledge was vague or important details were missing for multiple events or activities in the mosquito life cycle, such as the duration or location of the many periods when females rest between blood feeding and oviposition. The existing body of literature, though substantial, is not wholly reliable, and evidence for commonly held "facts" range from untraceable to extensive. Source references of some basic information are poor or date back more than 60 years, while other information that today is accepted widely as "fact" is not supported by evidence in the literature. Many topics, e.g., sugar feeding, resting preferences (location and duration), and blood feeding, merit being revisited in new geographical regions and ecological contexts to identify vulnerabilities for exploitation in control.

Extensive variation and strain-specificity in dengue virus susceptibility among African Aedes aegypti populations.

African populations of the mosquito Aedes aegypti are usually considered less susceptible to infection by human-pathogenic flaviviruses than globally invasive populations found outside Africa. Although this contrast has been well documented for Zika virus (ZIKV), it is unclear to what extent it is true for dengue virus (DENV), the most prevalent flavivirus of humans. Addressing this question is complicated by substantial genetic diversity among DENV strains, most notably in the form of four genetic types (DENV1 to DENV4), that can lead to genetically specific interactions with mosquito populations. Here, we carried out a continent-wide survey of DENV susceptibility using a panel of field-derived Ae. aegypti colonies from across the African range of the species and a colony from Guadeloupe, French West Indies as non-African reference. We found considerable variation in the ability of African Ae. aegypti populations to acquire and replicate a panel of six DENV strains spanning the four DENV types. Although African Ae. aegypti populations were generally less susceptible than the reference non-African population from Guadeloupe, in several instances some African populations were equally or more susceptible than the Guadeloupe population. Moreover, the relative level of susceptibility between African mosquito populations depended on the DENV strain, indicating genetically specific interactions. We conclude that unlike ZIKV susceptibility, there is no clear-cut dichotomy in DENV susceptibility between African and non-African Ae. aegypti. DENV susceptibility of African Ae. aegypti populations is highly heterogeneous and largely governed by the specific pairing of mosquito population and DENV strain.

Impact of physicochemical parameters of Aedes aegypti breeding habitats on mosquito productivity and the size of emerged adult mosquitoes in Ouagadougou City, Burkina Faso.

BACKGROUND: Outbreaks of dengue fever caused by viruses transmitted by Aedes aegypti mosquitoes are repeated occurrences in West Africa. In recent years, Burkina Faso has experienced major dengue outbreaks, most notably in 2016 and 2017 when 80% of cases were recorded in Ouagadougou City (Central Health Region). In order to better understand the ecology of this vector and to provide information for use in developing control measures, a study on the characteristics of Aedes container breeding sites and the productivity of such sites, as measured by the abundance of immature stages and resultant adult body size, was undertaken in three health districts (Baskuy, Bogodogo and Nongremassom) of Ouagadougou. METHODS: Adult mosquitoes were collected indoors and outdoors in 643 households during the rainy season from August to October 2018. The presence of water containers was systematically recorded and the containers examined for the presence or absence of larvae. Characteristics of the container breeding sites, including size of the container and temperature, pH and conductivity of the water contained within, were recorded as well as the volume of water. Traditional Stegomyia indices were calculated as quantitative indicators of the risk of dengue outbreaks; generalised mixed models were fitted to larval and pupal densities, and the contribution of each covariate to the model was evaluated by the Z-value and associated P-value. RESULTS: A total of 1061 container breeding sites were inspected, of which 760 contained immature stages of Ae. aegypti ('positive' containers). The most frequent container breeding sites found in each health district were tyres and both medium (buckets/cans/pots) and large (bins/barrels/drums) containers; these containers were also the most productive larval habitats and the types that most frequently tested positive. Of the Stegomyia indices, the Breteau, House and Container indices exceeded WHO dengue risk thresholds. Generalised linear mixed models showed that larval and pupal abundances were associated with container type, physicochemical characteristics of the water and collection month, but there were significant differences among container types and among health districts. Aedes aegypti body size was positively associated with type and diameter of the container, as well as with electrical conductivity of the water, and negatively associated with pH and temperature of the water and with the level of exposure of the container to sunlight. CONCLUSION: This study provides data on putative determinants of the productivity of habitats regarding Ae. aegypti immature stages. These data are useful to better understand Ae. aegypti proliferation. The results suggest that identifying and targeting the most productive container breeding sites could contribute to dengue vector control strategies in Burkina Faso.

First comprehensive analysis of Aedes aegypti bionomics during an arbovirus outbreak in west Africa: Dengue in Ouagadougou, Burkina Faso, 2016-2017.

BACKGROUND: Dengue's emergence in West Africa was typified by the Burkina Faso outbreaks in 2016 and 2017, the nation's largest to date. In both years, we undertook three-month surveys of Aedes populations in or near the capital city Ouagadougou, where the outbreaks were centered. METHODOLOGY: In 1200LG (urban), Tabtenga (peri-urban) and Goundry (rural) localities, we collected indoor and outdoor resting mosquito adults, characterized larval habitats and containers producing pupae and reared immature stages to adulthood in the laboratory for identification. All mosquito adults were identified morphologically. Host species (from which bloodmeals were taken) were identified by PCR. Generalized mixed models were used to investigate relationships between adult or larval densities and multiple explanatory variables. RESULTS: From samples in 1,780 houses, adult Ae. aegypti were significantly more abundant in the two urban localities (Tabtenga and 1200 LG) in both years than in the rural site (Goundry), where Anopheles spp. were far more common. Results from adult collections indicated a highly exophilic and anthropophilic (>90% bloodmeals of human origin) vector population, but with a relatively high proportion of bloodfed females caught inside houses. Habitats producing most pupae were waste tires (37% of total pupae), animal troughs (44%) and large water barrels (30%). While Stegomyia indices were not reliable indicators of adult mosquito abundance, shared influences on adult and immature stage densities included rainfall and container water level, collection month and container type/purpose. Spatial analysis showed autocorrelation of densities, with a partial overlap in adult and immature stage hotspots. CONCLUSION: Results provide an evidence base for the selection of appropriate vector control methods to minimize the risk, frequency and magnitude of future outbreaks in Ouagadougou. An integrated strategy combining community-driven practices, waste disposal and insecticide-based interventions is proposed. The prospects for developing a regional approach to arbovirus control in West Africa or across Africa are discussed.

Putative pleiotropic effects of the knockdown resistance (L1014F) allele on the life-history traits of Anopheles gambiae.

BACKGROUND: Existing mechanisms of insecticide resistance are known to help the survival of mosquitoes following contact with chemical compounds, even though they could negatively affect the life-history traits of resistant malaria vectors. In West Africa, the knockdown resistance mechanism kdrR (L1014F) is the most common. However, little knowledge is available on its effects on mosquito life-history traits. The fitness effects associated with this knockdown resistance allele in Anopheles gambiae sensu stricto (s.s.) were investigated in an insecticide-free laboratory environment. METHODS: The life-history traits of Kisumu (susceptible) and KisKdr (kdr resistant) strains of An. gambiae s.s. were compared. Larval survivorship and pupation rate were assessed as well as fecundity and fertility of adult females. Female mosquitoes of both strains were directly blood fed through artificial membrane assays and then the blood-feeding success, blood volume and adult survivorship post-blood meal were assessed. RESULTS: The An. gambiae mosquitoes carrying the kdrR allele (KisKdr) laid a reduced number of eggs. The mean number of larvae in the susceptible strain Kisumu was three-fold overall higher than that seen in the KisKdr strain with a significant difference in hatching rates (81.89% in Kisumu vs 72.89% in KisKdr). The KisKdr larvae had a significant higher survivorship than that of Kisumu. The blood-feeding success was significantly higher in the resistant mosquitoes (84%) compared to the susceptible ones (34.75%). However, the mean blood volume was 1.36 µL/mg, 1.45 µL/mg and 1.68 µL/mg in Kisumu, homozygote and heterozygote KisKdr mosquitoes, respectively. After blood-feeding, the heterozygote KisKdr mosquitoes displayed highest survivorship when compared to that of Kisumu. CONCLUSIONS: The presence of the knockdown resistance allele appears to impact the life-history traits, such as fecundity, fertility, larval survivorship, and blood-feeding behaviour in An. gambiae. These data could help to guide the implementation of more reliable strategies for the control of malaria vectors.

Interplay Between Oxytetracycline and the Homozygote kdr (L1014F) Resistance Genotype on Fecundity in Anopheles gambiae (Diptera: Culicidae) Mosquitoes.

The insecticide resistance in Anopheles gambiae mosquitoes has remained the major threat for vector control programs but the fitness effects conferred by these mechanisms are poorly understood. To fill this knowledge gap, the present study aimed at testing the hypothesis that antibiotic oxytetracycline could have an interaction with insecticide resistance genotypes and consequently inhibit the fecundity in An. gambiae. Four strains of An. gambiae: Kisumu (susceptible), KisKdr (kdr (L1014F) resistant), AcerKis (ace-1 (G119S) resistant) and AcerKdrKis (both kdr (L1014F) and ace-1 (G119S) resistant) were used in this study. The different strains were allowed to bloodfeed on a rabbit previously treated with antibiotic oxytetracycline at a concentration of 39·10-5 M. Three days later, ovarian follicles were dissected from individual mosquito ovaries into physiological saline solution (0.9% NaCl) under a stereomicroscope and the eggs were counted. Fecundity was substantially lower in oxytetracycline-exposed KisKdr females when compared to that of the untreated individuals and oxytetracycline-exposed Kisumu females. The exposed AcerKis females displayed an increased fecundity compared to their nontreated counterparts whereas they had reduced fecundity compared to that of oxytetracycline-exposed Kisumu females. There was no substantial difference between the fecundity in the treated and untreated AcerKdrKis females. The oxytetracycline-exposed AcerKdrKis mosquitoes had an increased fecundity compared to that of the exposed Kisumu females. Our data indicate an indirect effect of oxytetracycline in reducing fecundity of An. gambiae mosquitoes carrying kdrR (L1014F) genotype. These findings could be useful for designing new integrated approaches for malaria vector control in endemic countries.

Mosquito-Associated Viruses and Their Related Mosquitoes in West Africa.

Mosquito-associated viruses (MAVs), including mosquito-specific viruses (MSVs) and mosquito-borne (arbo)viruses (MBVs), are an increasing public, veterinary, and global health concern, and West Africa is projected to be the next front for arboviral diseases. As in-depth knowledge of the ecologies of both western African MAVs and related mosquitoes is still limited, we review available and comprehensive data on their diversity, abundance, and distribution. Data on MAVs' occurrence and related mosquitoes were extracted from peer-reviewed publications. Data on MSVs, and mosquito and vertebrate host ranges are sparse. However, more data are available on MBVs (i.e., dengue, yellow fever, chikungunya, Zika, and Rift Valley fever viruses), detected in wild and domestic animals, and humans, with infections more concentrated in urban areas and areas affected by strong anthropogenic changes. Aedes aegypti, Culex quinquefasciatus, and Aedes albopictus are incriminated as key arbovirus vectors. These findings outline MAV, related mosquitoes, key knowledge gaps, and future research areas. Additionally, these data highlight the need to increase our understanding of MAVs and their impact on host mosquito ecology, to improve our knowledge of arbovirus transmission, and to develop specific strategies and capacities for arboviral disease surveillance, diagnostic, prevention, control, and outbreak responses in West Africa.

Phenotypic Insecticide Resistance in Anopheles gambiae (Diptera: Culicidae): Specific Characterization of Underlying Resistance Mechanisms Still Matters.

An effective control of malaria vectors requires an extensive knowledge of mechanisms underlying the resistance-phenotypes developed by these vectors against insecticides. We investigated Anopheles gambiae mosquitoes from Benin and Togo for their intensity of insecticide resistance and we discussed the involvement of genotyped mechanisms in the resistance-phenotypes observed. Three- to five-day-old adult mosquitoes emerged from field and laboratory An. gambiae larvae were assayed using WHO tube intensity tests against various doses of deltamethrin: 1× (0.05%); 2× (0.1%); 5× (0.25%); 7.5× (0.375%) and those of pirimiphos-methyl: 0.5× (0.125%); 1× (0.25%). Members of An. gambiae complex were screened in field populations using polymerase chain reaction (PCR) assays. The presence of kdrR(1014F/1014S) and ace-1R(119S) mutations was also investigated using TaqMan and PCR-RFLP techniques, respectively. Anopheles gambiae from field were very resistant to deltamethrin, whereas KisKdr and AcerKdrKis strains displayed 100% mortality rates at 2× the diagnostic dose. In contrast, the field mosquitoes displayed a low resistance-intensity against 1× the diagnostic dose of pirimiphos-methyl, whereas AcerKis and AcerKdrKis strains showed susceptibility at 0.5× the diagnostic dose. Anopheles gambiae s.s., Anopheles coluzzii, and Anopheles arabiensis were identified. Allelic frequencies of kdrR (1014F) and ace-1R (119S) mutations in the field populations varied from 0.65 to 1 and 0 to 0.84, respectively. The field An. gambiae displayed high-resistance levels against deltamethrin and pirimiphos-methyl when compared with those of the laboratory An. gambiae-resistant strains. These results exhibit the complexity of underlying insecticide resistance mechanisms in these field malaria vectors.

Climate and Urbanization Drive Mosquito Preference for Humans.

The majority of mosquito-borne illness is spread by a few mosquito species that have evolved to specialize in biting humans, yet the precise causes of this behavioral shift are poorly understood. We address this gap in the arboviral vector Aedes aegypti. We first collect and characterize the behavior of mosquitoes from 27 sites scattered across the species' ancestral range in sub-Saharan Africa, revealing previously unrecognized variation in preference for human versus animal odor. We then use modeling to show that over 80% of this variation can be predicted by two ecological factors-dry season intensity and human population density. Finally, we integrate this information with whole-genome sequence data from 375 individual mosquitoes to identify a single underlying ancestry component linked to human preference. Genetic changes associated with human specialist ancestry were concentrated in a few chromosomal regions. Our findings suggest that human-biting in this important disease vector originally evolved as a by-product of breeding in human-stored water in areas where doing so provided the only means to survive the long, hot dry season. Our model also predicts that the rapid urbanization currently taking place in Africa will drive further mosquito evolution, causing a shift toward human-biting in many large cities by 2050.

Development and evaluation of a novel loop-mediated isothermal amplification (LAMP) method targeting Theileria parasites infecting Yezo sika deer.

The increasing Yezo sika deer (Cervus nippon yesoensis) population is creating a large problem. Yezo sika deer are an important blood meal source, and these deer contribute to the maintenance of tick populations. Theileria spp. infections in Yezo sika deer and T. orientalis infections in cows occur at high frequencies, and the same tick species infests both deer and cows. Therefore, a specific detection method to identify deer Theileria spp. is important. In this study, we establish a novel molecular detection method for identifying Theileria spp. from deer and tick samples using loop-mediated isothermal amplification (LAMP). This method targets a metalloprotease/cell division cycle protein gene homologue. Our LAMP protocol was able to detect deer Theileria and did not show cross reactivity with other closely related protozoan parasites, including T. orientalis. The LAMP method showed sensitivity and specificity equivalent to those of nested PCR performed on the same field samples from deer and ticks. These results demonstrate the applicability of LAMP to field surveys in which the detection of deer Theileria spp. is required. In conclusion, due to its simplicity, specificity, and reliability, we suggest our LAMP protocol as an appropriate method for routine surveys to detect Yezo sika deer and ticks infected with deer Theileria spp. parasites. Additionally, this LAMP method offers great promise as a useful tool to distinguish Yezo sika deer Theileria from related Theileria parasites present in livestock.

Genetic analysis and population structure of the Anopheles gambiae complex from different ecological zones of Burkina Faso.

The Anopheles gambiae complex (Diptera: Culicidae) is the most important vector for malaria in Sub-Saharan Africa, besides other vectors such as Anopheles funestus. Malaria vector control should encompass specific identification, genetic diversity and population structure of An. gambiae to design vector control strategies. The aim of this study was to determine the distribution of sibling species of the An. gambiae complex according to climatic regions related to cotton-growing or cotton-free areas by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Then, variation in mitochondrial cytochrome c oxidase 1 (COI) was used to assess the genetic structure within and between populations from our selected ecological zones. At the sibling species level, the following proportions were found across all samples (n = 180): An. coluzzii 65.56%, An. gambiae stricto sensu (s.s). 21.11%, and An. arabiensis 3.33%. Hybrids between An. gambiae s.s. and An. coluzzii (7.78%) and hybrids between An. coluzzii and An. arabiensis (2.22%) were found. The phylogenetic tree and Integer Neighbour-Joining (IntNJ) haplotype network did not reveal any distinct genetic structure pattern related to climatic or agricultural conditions in Burkina Faso. The Fst (Wright's F-statistic) values close to zero showed a free gene flow and no differentiation in An. gambiae complex populations. Furthermore, neutrality indices calculated by Tajima's D, Fu and Li's D⁎, Fu and Li's F⁎, Fu's Fs tests suggested an excess of rare mutations in the populations. Overall, variation in the proportions of An. gambiae s.s., An. coluzzii and An. arabiensis was found according to climatic regions, but COI analysis did not evidence any population structuring of the An. gambiae complex. These scientific contributions can be used as a basis for further in-depth study of the genetic diversity of the An. gambiae complex for epidemiological risk assessment of malaria in Burkina Faso.

Proteomic Investigation on Anopheles gambiae in Burkina Faso Related to Insecticide Pressures from Different Climatic Regions.

In Sub-Saharan Africa, An. gambiae sensu lato (s.l.) Giles 190, largely contributes to malaria transmission. Therefore, the authors carry out a proteomic analysis to compare its metabolic state, depending on different pesticide pressures by selecting areas with/without cotton crops. The proteomes data are available via ProteomeXchange with identifier PXD016300. From a total of 1.182 identified proteins, 648 are retained for further statistical analysis and are attributed to biological functions, the most important of which being energy metabolism (120 proteins) followed by translation-biogenesis (74), cytoskeleton (71), stress response (62), biosynthetic process (60), signalling (44), cellular respiration (38), cell redox homeostasis (25), DNA processing (17), pheromone binding (10), protein folding (9), RNA processing (9), other proteins (26) and unknown functions (83). In the Sudano-Sahelian region, 421 (91.3%) proteins are found in samples from areas both with and without cotton crops. By contrast, in the Sahelian region, only 271 (55.0%) are common to both crop areas, and 233 proteins are up-regulated from the cotton area. The focus is placed on proteins with putative roles in insecticide resistance, according to literature. This study provides the first whole-body proteomic characterisation of An. gambiae s.l. in Burkina Faso, as a framework to strengthen vector control strategies.