Distinct physiological, biochemical and morphometric adjustments in the malaria vectors Anopheles gambiae and A. coluzzii as means to survive dry season conditions in Burkina Faso.

Aestivation and dispersive migration are the two strategies evoked in the literature to explain the way in which malaria vectors Anopheles coluzzii and A. gambiae survive the harsh climatic conditions of the dry season in sub-Saharan Africa. However, the physiological mechanisms regulating these two strategies are unknown. In the present study, mosquito species were exposed to controlled environmental conditions mimicking the rainy and dry seasons of south western Burkina Faso. Survival strategies were studied through morphometric (wing length), ecophysiological (respiratory gas exchanges), biochemical (cuticular hydrocarbons composition) and molecular (AKH mRNA expression levels) parameters, variations of which are usually considered to be hallmarks of aestivation and dispersion mechanisms in various insects. Our results showed that ecophysiological and morphometric adjustments are made in both species to prevent water losses during the dry season. However, the usual metabolic rate modifications expected as signatures of aestivation and migration were not observed, highlighting specific and original physiological mechanisms sustaining survival in malaria mosquitoes during the dry season. Differences in epicuticular hydrocarbon composition and AKH levels of expression were found between the permanent and temporary A. coluzzii populations, illustrating the great phenotypic plasticity of this mosquito species. Altogether, our work underlines the diverse and complex pattern of changes occurring in the two mosquito species and at the population level to cope with the dry season and highlights potential targets of future control tools.

Monitoring Dry Season Persistence of Anopheles gambiae s.l. Populations in a Contained Semi-Field System in Southwestern Burkina Faso, West Africa.

To gain insight into the dry season survival strategies of Anopheles gambiae s.l., a new contained semi-field system was developed and used for the first time in Burkina Faso, West Africa. The system consisted of a screened greenhouse within which the local environment was reproduced, including all ecological requirements for mosquito development cycle completion. The system was seeded with the progenies of female Anopheles gambiae, Anopheles coluzzii, and Anopheles arabiensis collected in the vicinity of the greenhouse during the rainy season. After successful establishment in the semi-field system, mosquito populations were monitored over a 1-yr period by regular surveys of larval and adult specimens. We provided evidence for the persistence of adult mosquitoes throughout the dry season, in the absence of any suitable larval development site. During the hot and dry periods, adult insects were observed in artificial shelters (clay pots, building blocks, and dark corners). The mosquito population rapidly built up with the return of the rainy season in the area, when artificial breeding sites were refilled in the enclosure. However, only An. coluzzii and, later, An. arabiensis were detected in the subsequent rainy season, whereas no An. gambiae specimen was found. Our findings suggest that An. coluzzii and An. arabiensis may be able to aestivate throughout the dry season in Southwestern Burkina Faso, whereas An. gambiae might adopt a different dry-season survival strategy, such as long-distance re-colonization from distant locations. These results may have important implications for malaria control through targeted vector control interventions.

Impaired fatty acid oxidation as a cause for lipotoxicity in cardiomyocytes.

A major cause for diabetic cardiomyopathy is excess lipid accumulation. To elucidate mechanisms of lipotoxicity mediated diabetic heart disease we need to further our understanding of how lipid metabolism is altered in the diabetic heart. Here we investigated the role of lipid clearance by oxidation as a regulator of lipid-mediated toxicity (lipotoxicity). We evaluated the effect of pre-treating rat neonatal cardiomyocytes (NCMs) with either oleate (mono-unsaturated fatty acid) or palmitate (saturated fatty acid) on fatty acid oxidation (FAO) by measuring (14)C-CO2 production. We evaluated carnitine palmitoyltransferase (Cpt1b) expression by western blotting and mitochondrial membrane potential by quantitative and qualitative fluorescence analyses using the JC-1 dye. We inhibited the Cpt1b pharmacologically using etomoxir and genetically by knocking down its expression using LentiVector mediated transduction of siRNAs targeting the Cpt1b gene. We found that palmitate had a slower clearance rate from NCMs than oleate, and this was associated with a significant decrease in FAO. This impairment in FAO was not the result of either loss of Cpt1b protein or mitochondrial integrity. Enhancing FAO with either oleate or carnitine was associated with a significant attenuation of palmitate mediated lipotoxicity. In contrast impairing FAO in oleate treated NCMs caused lipotoxicity. Here we demonstrate that a major difference between non-toxic unsaturated fatty acids and toxic saturated fatty acids is there ability to stimulate or inhibit fatty acid oxidation, respectively. This has important implications for diabetic cardiomyopathy since diabetic hearts consistently exhibit elevated lipid accumulation.

Polymorphic chromosomal inversions in Anopheles moucheti, a major malaria vector in Central Africa.

Anopheles moucheti Evans (Diptera: Culicidae) is a major vector of malaria in forested areas of Central Africa. However, few genetic tools are available for this species. The present study represents the first attempt to characterize chromosomes in An. moucheti females collected in Cameroon. Ovarian nurse cells contained polytene chromosomes, which were suitable for standard cytogenetic applications. The presence of three polymorphic chromosomal inversions in An. moucheti was revealed. Two of these inversions were located on the 2R chromosome arm. The homology between the 2R chromosome arms of An. moucheti and Anopheles gambiae Giles was established by fluorescent in situ hybridization of six An. gambiae genic sequences. Mapping of the probes on chromosomes of An. moucheti detected substantial gene order reshuffling between the two species. The presence of polytene chromosomes and polymorphic inversions in An. moucheti provides a new basis for further population genetic, taxonomic and ecological studies of this neglected malaria vector.

Assortative mating in mixed swarms of the mosquito Anopheles gambiae s.s. M and S molecular forms, in Burkina Faso, West Africa.

The molecular form composition of Anopheles gambiae Giles s.s. (Diptera: Culicidae) mating swarms and the associated mating pairs (copulae) were investigated during two rainy seasons (July to October, 2005 and July to November, 2006) in the villages of Soumousso and Vallée du Kou (VK7). Although the habitats of these villages differ markedly, sympatric populations of M and S molecular forms of An. gambiae s.s. occur in both places periodically. The main aim was to assess the degree to which these molecular forms mate assortatively. In Soumousso, a wooded savannah habitat, the majority of swarm samples consisted of only S-form males (21/28), although a few M-form males were found in mixed M- and S-form swarms. In VK7, a rice growing area, the majority of swarm samples consisted of only M-form males (38/62), until October and November 2006, when there were nearly as many mixed-form as single-form swarms. Overall, ∼60% of M- and S-form swarms were temporally or spatially segregated; the two forms were effectively prevented from encountering each other. Of the remaining 40% of swarms, however, only about half were single-form and the rest were mixed-form. Of the 33 copulae collected from mixed-form swarms, only four were mixed-form pairs, significantly fewer than expected by random pairing between forms (χ(2) = 10.34, d.f. = 2, P < 0.01). Finally, all specimens of inseminated females were of the same form as the sperm contained within their spermatheca (n = 91), even for the four mixed-form copulae. These findings indicate that assortative mating occurs within mixed-form swarms, mediated most probably by close-range mate recognition cues.

Larval habitat segregation between the molecular forms of the mosquito Anopheles gambiae in a rice field area of Burkina Faso, West Africa.

In West Africa, lineage splitting between the M and S molecular forms of the major Afro-tropical malaria mosquito, Anopheles gambiae (Diptera: Culicidae), is thought to be driven by ecological divergence, occurring mainly at the larval stage. Here, we present evidence for habitat segregation between the two molecular forms in and around irrigated rice fields located within the humid savannahs of western Burkina Faso. Longitudinal sampling of adult mosquitoes emerging from a range of breeding sites distributed along a transect extending from the heart of the rice field area into the surrounding savannah was conducted from June to November 2009. Analysis revealed that the two molecular forms and their sibling species Anopheles arabiensis are not randomly distributed in the area. A major ecological gradient was extracted in relation to the perimeter of the rice fields. The M form was associated with larger breeding sites mostly consisting of rice paddies, whereas the S form and An. arabiensis were found to depend upon temporary, rain-filled breeding sites. These results support hypotheses about larval habitat segregation and confirm the suggestion that the forms have different larval habitat requirements. Segregation appears to be clearly linked to anthropogenic permanent habitats and the community structure they support.

Physiology and development of the M and S molecular forms of Anopheles gambiae in Burkina Faso (West Africa).

In West Africa, M and S molecular forms of Anopheles gambiae sensu stricto (Diptera: Culicidae) Giles, frequently occur together, although with different population bionomics. The S form typically breeds in rain-dependant water collections and is present during the rainy season only whereas the M form can thrive all year long in areas with permanent breeding opportunities. In the present study, we explored physiological and developmental trade-offs at play in laboratory colonies and field populations of the M and S forms that originated from an area of sympatry in Burkina Faso, where M and S larvae exhibit such habitat segregation. In the laboratory, larvae of the M form developed slower than the S form (mean values 9.51 and 8.85 days, respectively, Wilcoxon's test, P < 0.001). Although wing length and dry weight at emergence showed large variations, M females were on average 8% heavier than S females of similar wing length. Higher nutritional reserves (proteins and lipids) in teneral adults explained part of this weight difference, reflecting a better ability of the M form to garner resources at the larval stage. Furthermore, a higher rate of ovarian maturation was observed in the M form after a single bloodmeal. The relevance of these findings for parasite transmission is discussed.

Optimization of breeding output for larval stage of Anopheles gambiae (Diptera: Culicidae): prospects for the creation and maintenance of laboratory colony from wild isolates.

Domesticating anopheline species from wild isolates provides an important laboratory tool but requires detailed knowledge of their natural biology and ecology, especially the natural breeding habitats of immature stages. The aim of this study was to determine the optimal values of some parameters of Anopheles gambiae larval development, so as to design a standard rearing protocol of highland isolates, which would ensure: the biggest fourth instars, the highest pupae productivity, the shortest duration of the larval stage and the best synchronization of pupation. The density of larvae, the size of breeding water and the quantity of food supplied were tested for their effect on larval growth. Moreover, three cheap foodstuffs were selected and tested for their capability to improve the breeding yield versus TetraMin® as the standard control. The larval density was a very sensitive parameter. Its optimal value, which was found to be ≈1 cm-2 surface area, yielded a daily pupation peak of 38.7% on day 8 post-oviposition, and a global pupae productivity of 78.7% over a duration range of three days. Anopheles gambiae's larval growth, survival and developmental synchronization were density-dependent, and this species responded to overcrowding by producing smaller fourth instars and fewer pupae, over elongated immature lifetime and duration range of pupae occurrence, as a consequence of intraspecific competition. While shallow breeding waters (<3 cm) produced a higher number of pupae than deeper ones, no effect of the breeding habitat's absolute surface area on larval development was observed. Increasing the daily food supply improved the pupae productivity but also boosted the water pollution level (which was assessed by the biological oxygen demand (BOD) and the chemical oxygen demand (COD)) up to a limit depending on the food quality, above which a rapid increase in larval mortality was recorded. The food quality that could substitute the manufactured baby fish food was obtained with weighed mixture of 1 wheat+1 shrimp+2 fish. On establishing an anopheline mosquito colony in the laboratory, special care should be taken to design and maintain the appropriate optimal values of larval density, water depth, daily diet quantity and nutritional quality.

[Preliminary evaluation of the insecticide susceptibility in Anopheles gambiae and Culex quinquefasciatus from Lobito (Angola), using WHO standard assay]. / Premiers tests OMS d'évaluation de la sensibilité aux insecticides chez Anopheles gambiae et Culex quinquefasciatus à Lobito, Angola.

Field collections of the most common urban mosquito vectors Anopheles gambiae and Culex quinquefasciatus were carried out in June 2003, March 2004 and November 2005 to gather preliminary data on the insecticide susceptibility in mosquitoes from Lobito (Angola) using the WHO standard bioassays. Bioassays were performed on F0 adults emerging from the field larval collections and on unfed adults from landing catches on volunteers. Batches of mosquitoes from three selected locations (Alto Liro, San Jao and Bela Vista) were exposed for 1 hour to several insecticides such as DDT 4%, carbosulfan 0.4%, permethrin 1%, deltamethrin 0.05% and cyfluthrin 0.15%, in order to estimate the immediate knockdown times (kdT50 and kdT95) and the mortality rate after exposure. The results revealed that mosquito susceptibility to insecticides varied depending on the insecticide, the site and the period of collection. The main local malaria vector A. gambiae (both M and S forms) was basically resistant to DDT and susceptible to all pyrethoids, regardless of the period and the site of collections. The overall mortality rate due to DDT was 73% in Alto Liro, 89% in San Jao and varied depending on the period in Bela Vista between 95% in March 2004 and 100% in November 2005. The mortality due to pyrethoids was 100% at all locations, with the kdT50 and KdT95 times ranging between 9 and 16 minutes and between 18 and 29 minutes, respectively. Concerning the C. quinquefasciatus, populations from Yard and Caponte were resistant to all insecticides tested; the mortality rate was 40% with deltamethrin and 70% with permethrin, while no lethal effect was observed with DDT or carbosulfan. In conclusion, despite its probable high resistance to DDT, the main local malaria vector A. gambiae remained fully susceptible to pyrethroids. This could forecast a good biological efficacy of the scheduled vector control interventions in Angola, based on a large-scale distribution of long-lasting, insecticide-treated nets and on the implementation of indoor residual spraying. The local vector control programme must include well-adapted IEC campaigns and full participation of the community for better management of the insecticide resistance in targeted mosquitoes and for better control of malaria vector populations.

Genome-wide analysis of transcriptomic divergence between laboratory colony and field Anopheles gambiae mosquitoes of the M and S molecular forms.

Our knowledge of Anopheles gambiae molecular biology has mainly been based on studies using inbred laboratory strains. Differences in the environmental exposure of these and natural field mosquitoes have inevitably led to physiological divergences. We have used global transcript abundance analyses to probe into this divergence, and identified transcript abundance patterns of genes that provide insight on specific adaptations of caged and field mosquitoes. We also compared the gene transcript abundance profiles of field mosquitoes belonging to the two morphologically indistinguishable but reproductively isolated sympatric molecular forms, M and S, from two different locations in the Yaoundé area of Cameroon. This analysis suggested that environmental exposure has a greater influence on the transcriptome than does the mosquito's molecular form-specific genetic background.

Geographic and ecological distribution of the dengue and chikungunya virus vectors Aedes aegypti and Aedes albopictus in three major Cameroonian towns.

Aedes albopictus (Diptera: Culicidae) was first reported in Central Africa in 2000, together with the indigenous mosquito species Aedes aegypti (Diptera: Culicidae). Because Ae. albopictus can also transmit arboviruses, its introduction is a public health concern. We undertook a comparative study in three Cameroonian towns (Sahelian domain: Garoua; equatorial domain: Douala and Yaoundé) in order to document infestation by the two species and their ecological preferences. High and variable levels of pre-imaginal Ae. aegypti and Ae. albopictus infestation were detected. Only Ae. aegypti was encountered in Garoua, whereas both species were found in Douala and Yaoundé, albeit with significant differences in their relative prevalence. Peridomestic water containers were the most strongly colonized and productive larval habitats for both species. No major differences in types of larval habitat were found, but Ae. albopictus preferentially bred in containers containing plant debris or surrounded by vegetation, whereas Ae. aegypti tended to breed in containers located in environments with a high density of buildings. These findings may have important implications for vector control strategies.

Changes in malaria epidemiology in France and worldwide, 2000-2015.

In 2015, 212 million new cases of malaria were reported, causing 429,000 deaths. The World Health Organization (WHO) estimated a 41% decrease in the number of new cases worldwide between 2000 and 2015. The number of deaths from malaria fell by 62% worldwide and by 71% in Africa. In mainland France, malaria is mainly imported by travelers or migrants from endemic areas, in particular sub-Saharan Africa (95%). In France, the number of imported malaria cases, mainly due to Plasmodium falciparum (85%), was estimated at about 82,000 for the period 2000-2015. Over the same period, 6,468 cases of malaria were reported in the French armed forces, of which 2,430 cases (37.6%) were considered as imported because occurring outside of endemic areas. The number of malaria cases also fell between 2000 and 2015 in Mayotte and French Guiana, a malaria transmission zone. Mayotte has entered the elimination of malaria with less than 15 cases per year. In French Guiana, between 300 and 500 cases have been reported annually in recent years. The decline in morbidity and mortality is usually attributed to vector control measures and improved access to effective treatments. However, the Anopheles mosquitoes that transmit the disease have developed resistance against most insecticides. Similarly, malaria parasites have developed resistance against most of the antimalarial drugs used as prevention or treatment, even the latest marketed combinations such as artemisinin-based combination therapies.

Distribution of insensitive acetylcholinesterase (ace-1R) in Anopheles gambiae s.l. populations from Burkina Faso (West Africa).

OBJECTIVE: To investigate through countrywide sampling at 20 localities across the three different agro-climatic zones of Burkina Faso, the distribution of the acetylcholinesterase insensitive mutation ace-1(R), which confers resistance to organophosphates (OP) and carbamates (CM) insecticides in An. gambiae s.l. METHODS: Adult mosquitoes were collected by indoor aerosol spraying from August to October 2006. Specimens were identified to species by polymerase chain reaction (PCR) assay and characterized for the ace-1(R) mutation using a PCR-restriction fragment length polymorphism diagnostic. RESULTS: Collected mosquitoes were a mixture of An. gambiae s.s. and An. arabiensis across the Sudan (98.3%vs. 1.7%), Sudan-sahelian (78.6%vs. 21.4%) and the Sahel (91.5%vs. 8.5%) ecotypes. The An. gambiae S-form predominated in the Sudan sites from the West (69%vs. 31% for the M form) but was not found in the Sahel (100% M form). The ace-1(R) mutation was dispersed throughout the Sudan and Sudan-sahelian localities at moderate frequency (<50%) but was absent in the Sahel. It was far more prevalent in S form than M form mosquitoes (0.32 for the S form vs. 0.036 for the M form). No An. arabiensis was detected carrying the mutation. The geographic distribution of ace-1(R) in the Sudan and Sudan-sahelian correlated with the cotton growing areas dispersed throughout the two climatic zones. CONCLUSIONS: These results have special significance as OP and CM insecticides have been proposed as alternatives or additions to pyrethroids which are currently used exclusively in many vector control programmes.

Dynamics of insecticide resistance in the malaria vector Anopheles gambiae s.l. from an area of extensive cotton cultivation in Northern Cameroon.

OBJECTIVE: To explore temporal variation in insecticide susceptibility of Anopheles gambiae s.l. populations to the four chemical groups of insecticides used in public health and agriculture, in close match with the large-scale cotton spraying programme implemented in the cotton-growing area of North Cameroon. METHODS: Mosquito larvae were collected in 2005 before (mid June), during (mid August) and at the end (early October) of the cotton spraying programme. Larvae were sampled in breeding sites located within the cotton fields in Gaschiga and Pitoa, and in Garoua, an urban cotton-free area that served as a control. Insecticide susceptibility tests were carried out with 4% DDT (organochlorine), 0.4% chlorpyrifos methyl (organophosphate), 0.1% propoxur (carbamate), 0.05% deltamethrin and 0.75% permethrin (pyrethroids). RESULTS: Throughout the survey, An. gambiae s.l. populations were completely susceptible to carbamate and organophosphate, whereas a significant decrease of susceptibility to organochlorine and pyrethroids was observed during spraying in cotton-growing areas. Tolerance to these insecticides was associated with a slight increase of knockdown times compared to the reference strain. Among survivor mosquitoes, the East and West African Kdr mutations were detected only in two specimens of An. gambiae s.s. (n = 45) and not in Anopheles arabiensis (n = 150), suggesting metabolic-based resistance mechanisms. CONCLUSIONS: Environmental disturbance due to the use of insecticides in agriculture may provide local mosquito populations with the enzymatic arsenal selecting tolerance to insecticides.

Comparative physiological plasticity to desiccation in distinct populations of the malarial mosquito Anopheles coluzzii.

BACKGROUND: In West Africa, populations of the malaria vector mosquito, Anopheles coluzzii, are seasonally exposed to strong desiccating conditions during the dry season. Their dynamics strictly follows the pace of the availability of suitable larval development sites (water collections). Accordingly, mosquitoes can reproduce all year long where permanent breeding is possible, or stop reproduction and virtually disappear at the onset of the dry season when surface water dries up, like observed in temporary habitats of dry savannah areas. This highlights the strong adaptive abilities of this mosquito species, which relies at least in part, upon physiological and molecular mechanisms of specific signatures. METHODS: Here, we analysed a range of physiological and molecular responses expressed by geographically different populations of An. coluzzii inhabiting permanent and temporary breeding sites from the north and the south-west of Burkina Faso. Four mosquito colonies, namely (i) Oursi, built from females breeding in permanent habitats of the north; (ii) Déou, from temporary northern habitats; (iii) Soumousso from south-western temporary breeding sites; and (iv) Bama, from permanent habitats of the same south-western zone, were reared in climatic chambers under contrasted environmental conditions, mimicking temperature, relative humidity and light regimen occurring in northern Burkina Faso. Female mosquitoes were analysed for the seasonal variation in their amounts of proteins, triglycerides and free-circulating metabolites. The expression level of genes coding for the adipokinetic (AKH-I) and the AKH/corazonin-related peptides (ACP) were also assessed and compared among populations and environmental conditions. RESULTS: Our analysis did not reveal an apparent pattern of physiological and molecular variations strictly correlated with either the larval ecotype or the geographical origin of the mosquitoes. However, specific distinct responses were observed among populations, suggesting that dry season survival may rely on more complex ecological parameters at a micro-habitat scale. Interestingly, the physiological and molecular data support the hypothesis that different aestivation abilities exist among populations of An. coluzzii inhabiting contrasted ecological settings. In particular, the striking metabotypes differentiation and the AKH mRNA expression level observed in females from temporary northern populations may suggest the existence of a "strong" aestivation strategy in these specimens. CONCLUSION: Our work provides insights into the physiological and molecular basis of dry and rainy season responses in An. coluzzii, and highlights the important diversity of the mechanisms involved. Such results represent key data for understanding the ecophysiological mechanisms underpinning the strong adaptive potential of this malaria vector species, which undoubtedly contributes to the spreading of mosquito distribution areas in space and time.

[Malaria vectors: from the field to genetics. Research in Africa]. / Vecteurs de paludisme: du terrain à la génétique moléculaire. Recherches en Afrique.

Only about 60 Anopheline species transmit malaria among more than 3,000 mosquito species recorded in the world. In Africa, the major vectors are Anopheles gambiae,An. arabiensis, An. funestus, An. nili and An. moucheti. They all belong to species complexes or groups of closely related species that are very difficult to set apart on morphological grounds, but which may have highly variable behaviours and vectorial capacities. Understanding this complexity is of major importance in vector control programs or for implementing any public health intervention program such as drugs or vaccine trials. Among the seven species of the complex,Anopheles gambiaes.s. shows a huge chromosomal polymorphism related to adaptation to specific natural or anthropic environments, from equatorial forested Africa to dry sahelian areas. Recent studies conducted in West and Central Africa suggest an incipient speciation into 2 molecular forms provisionally called M and S. A similar evolutionary phenomenon is observed in An. funestus, in which sympatric populations carrying specific chromosomal paracentric inversions showed restricted gene flow. Distribution of species from An. nili group and An. moucheti complex is restricted to more humid regions of Africa. However in some areas these species play the major role in malaria transmission. Comprehensive knowledge of transmission cycles and of behavioural and underlying genetic heterogeneities that exist within and among natural vector populations will thus benefit the whole area of malaria control and epidemiology. Molecular and genetic studies, as well as in depth monitoring of vector biology, have been recently facilitated by advances in functional and comparative genomics, including recent publication of the nearly complete genome sequence of An. gambiae. Challenge for the next years is to answer to the very simple question: why is an insect a vector?

[Entomological factors of arboviruses emergences]. / Facteurs entomologiques d'émergence des arboviroses.

Arboviruses - viruses transmitted by haematophagous arthropods - are responsible for febrile syndromes, which sometimes include haemorrhagic or neurological symptoms. Human activities have facilitated the emergence of these originally zoonotic viruses and the domestication and spread throughout the world of their major vectors. The last decade has seen significant changes in the epidemiology of arboviruses transmitted by mosquitoes of the genus Aedes, particularly in relation to the intercontinental spread of Aedes albopictus. Here, we address the epidemiological consequences of the invasion by this species into Central Africa and Europe in a context of viral globalization. The risk of transmission in these areas is influenced by virus-vector adaptation phenomena as well as environmental phenomena including climate. Faced with these new risks, it is essential to develop competences in entomological and virological surveillance, risk assessment and forecasting of epidemic risk in order to develop strategies for the prevention and control of epidemics.

Combining two-dimensional gel electrophoresis and metabolomic data in support of dry-season survival in the two main species of the malarial mosquito Anopheles gambiae.

In dry savannahs of West-Africa, the malarial mosquitoes of the Anopheles gambiae sensu stricto complex annually survive the harsh desiccating conditions of the dry season. However, the physiological and biochemical mechanisms underlying how these mosquitoes survive such desiccating conditions are still undefined, and controversial. In this context, we provide the first work examining both proteomic and metabolomic changes in the two molecular forms of A. gambiae s.s (M and S forms) experimentally exposed to the rainy and dry season conditions as they experience in the field. Protein abundances of the mosquitoes were measured using a two-dimensional fluorescence difference gel electrophoresis (2D DIGE) coupled with a matrix-assisted laser desorption/ionisation-time of flight (MALDI-TOF) and tandem mass spectrometry (MS) for protein identification. These assays were conducted by Applied Biomics (http://www.appliedbiomics.com, Applied Biomics, Inc. Hayward, CA, USA), and the mass spectrometry proteomics data have been deposited to the ProteomeXchange Consortium (http://proteomecentral.proteomexchange.org) via the PRIDE partner repository with the dataset identifier PXD000294. The metabolomic analysis was conducted using both Acquity UPLC(®) system (for amino acid identification), and a gas-chromatography-mass spectrometry platform (for sugars identification). Metabolomic fingerprintings were assessed in the University of Rennes 1, UMR CNRS 6553 EcoBio (France). A detailed interpretation of the obtained data can be found in Hidalgo et al. (2014) [1] (Journal of Insect Physiology (2014)).

Cystic fibrosis mutations in French Canadians: three CFTR mutations are relatively frequent in a Quebec population with an elevated incidence of cystic fibrosis.

The French-Canadian population in the Saguenay-Lac St. Jean region of northeastern Quebec has an elevated frequency of cystic fibrosis (CF). The average incidence of cystic fibrosis was 1 in 891 births and the prevalence of CF carriers was estimated to be 1 in 15. We tested for 10 mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) gene in 133 French-Canadian CF families from Quebec. Ninety-one families were from the Saguenay-Lac St. Jean region and 42 families were referred from other regions of Quebec. We detected the CFTR mutation in 93 and 92% of the CF chromosomes in the Saguenay-Lac St. Jean and the major-urban Quebec families, respectively. The two groups of French-Canadian families were significantly different for the proportions of CFTR mutations. The three most common mutations in the Saguenay-Lac St. Jean families were delta F508 (58%), 621 + 1G----T (23%), and A455E (8%); and in the major-urban Quebec families were delta F508 (71%), 711 + 1G----T (9%), and 621 + 1G----T (5%). These results provide evidence for the role of founder effect in the elevated incidence of cystic fibrosis in the Saguenay-Lac St. Jean population.

High amounts of genetic differentiation between populations of the malaria vector Anopheles arabiensis from West Africa and eastern outer islands.

Polymorphism at nine microsatellite loci was examined to assess the level of genetic differentiation between four Anopheles arabiensis populations from Senegal, the high plateau of Madagascar, and Reunion and Mauritius islands. Eight of nine loci showed great polymorphism (2-16 alleles/locus) and significant genetic differentiation was revealed between all four populations by F- and R-statistics, with Fst estimates ranging from 0.080 to 0.215 and equivalent Rst values ranging between 0.022 and 0.300. These high amounts of genetic differentiation are discussed in relation to geographic distance including large bodies of water, and history of mosquito settlement, and insecticide use on the islands. The results suggest that historical events of drift rather than mutation are probably the forces generating genetic divergence between these populations, with homogenization of the gene pool by migration being drastically restricted across the ocean.