Ibrahima Dia

Distribution of knock-down resistance mutations in Anopheles gambiae molecular forms in west and west-central Africa

BackgroundKnock-down resistance (kdr) to DDT and pyrethroids in the major Afrotropical vector species, Anopheles gambiae sensu stricto, is associated with two alternative point mutations at amino acid position 1014 of the voltage-gated sodium channel gene, resulting in either a leucine-phenylalanine (L1014F), or a leucine-serine (L1014S) substitution. In An. gambiae S-form populations, the former mutation appears to be widespread in west Africa and has been recently reported from Uganda, while the latter, originally recorded in Kenya, has been recently found in Gabon, Cameroon and Equatorial Guinea. In M-form populations surveyed to date, only the L1014F mutation has been found, although less widespread and at lower frequencies than in sympatric S-form populations.MethodsAnopheles gambiae M- and S-form specimens from 19 sites from 11 west and west-central African countries were identified to molecular form and genotyped at the kdr locus either by Hot Oligonucleotide Ligation Assay (HOLA) or allele-specific PCR (AS-PCR).ResultsThe kdr genotype was determined for about 1,000 An. gambiae specimens. The L1014F allele was found at frequencies ranging from 6% to 100% in all S-form samples (N = 628), with the exception of two samples from Angola, where it was absent, and coexisted with the L1014S allele in samples from Cameroon, Gabon and north-western Angola. The L1014F allele was present in M-form samples (N = 354) from Benin, Nigeria, and Cameroon, where both M- and S-forms were sympatric.ConclusionThe results represent the most comprehensive effort to analyse the overall distribution of the L1014F and L1014S mutations in An. gambiae molecular forms, and will serve as baseline data for resistance monitoring. The overall picture shows that the emergence and spread of kdr alleles in An. gambiae is a dynamic process and that there is marked intra- and inter-form heterogeneity in resistance allele frequencies. Further studies are needed to determine: i) the importance of selection pressure exerted by both agricultural and public health use of pyrethroid insecticides, ii) the phenotypic effects, particularly when the two mutations co-occur; and iii) the epidemiological importance of kdr for both pyrethroid- and DDT-based malaria control operations, particularly if/when the two insecticides are to be used in concert.

Anopheles gambiae complex along The Gambia river, with particular reference to the molecular forms of An. gambiae s.s.

BackgroundThe geographic and temporal distribution of M and S molecular forms of the major Afrotropical malaria vector species Anopheles gambiae s.s. at the western extreme of their range of distribution has never been investigated in detail.Materials and methodsCollections of indoor-resting An. gambiae s.l. females were carried out along a ca. 400 km west to east transect following the River Gambia from the western coastal region of The Gambia to south-eastern Senegal during 2005 end of rainy season/early dry season and the 2006 rainy season. Specimens were identified to species and molecular forms by PCR-RFLP and the origin of blood-meal of fed females was determined by ELISA test.ResultsOver 4,000 An. gambiae s.l. adult females were collected and identified, 1,041 and 3,038 in 2005 and 2006, respectively. M-form was mainly found in sympatry with Anopheles melas and S-form in the western part of the transect, and with Anopheles arabiensis in the central part. S-form was found to prevail in rural Sudan-Guinean savannah areas of Eastern Senegal, in sympatry with An. arabiensis. Anopheles melas and An. arabiensis relative frequencies were generally lower in the rainy season samples, when An. gambiae s.s. was prevailing. No large seasonal fluctuations were observed for M and S-forms. In areas where both M and S were recorded, the frequency of hybrids between them ranged from to 0.6% to 7%.DiscussionThe observed pattern of taxa distribution supports the hypothesis of a better adaptation of M-form to areas characterized by water-retaining alluvial deposits along the Gambia River, characterized by marshy vegetation, mangrove woods and rice cultivations. In contrast, the S-form seems to be better adapted to free-draining soil, covered with open woodland savannah or farmland, rich in temporary larval breeding sites characterizing mainly the eastern part of the transect, where the environmental impact of the Gambia River is much less profound and agricultural activities are mainly rain-dependent. Very interestingly, the observed frequency of hybridization between the molecular forms along the whole transect was much higher than has been reported so far for other areas.ConclusionThe results support a bionomic divergence between the M and S-forms, and suggest that the western extreme of An. gambiae s.s. geographical distribution may represent an area of higher-than-expected hybridization between the two molecular forms.

Vector competence of Aedes aegypti populations from Senegal for sylvatic and epidemic dengue 2 virus isolated in West Africa

Vector competence of Aedes aegypti populations from six locations in different bioclimatic zones and habitats of Senegal was assessed for sylvatic (ArD 140875) and epidemic (ArA 6894) dengue 2 virus (DENV-2) isolates. F1 generation mosquitoes were fed with a blood meal containing 10(6.5) and 1.6 x 10(7) tissue culture infectious dose (TCID50) of the sylvatic DENV-2 isolate or 10(6.5) TCID50 of the epidemic isolate and incubated for 14 days. After this period, legs and wings of each individual were pooled and the corresponding body was collected separately. An immunofluorescence assay was used to test bodies and wings/legs for DENV-2 infection separately. The results showed that: (i) Senegalese Ae. aegypti populations have lower vector competence (infection rate 0-26%, dissemination rate 10-100%) for sylvatic and epidemic dengue viruses than populations from Asia and America; (ii) there was no obvious geographic variation in susceptibility among mosquito populations; (iii) the only apparent factor affecting the susceptibility was the virus titre; and (iv) except for that of Kédougou, all populations were able to disseminate the virus. Overall our findings, while extended to four other populations, confirm previous studies showing low susceptibility of Senegalese Ae. aegypti populations.

Bionomics of Anopheles gambiae Giles, An. arabiensis Patton, An. funestus Giles and An. nili (Theobald) (Diptera: Culicidae) and Transmission of Plasmodium falciparum in a Sudano-Guinean Zone (Ngari, Senegal)

Abstract An entomological study was conducted in a village of Sudano-Guinean savanna in Senegal, during the rainy season from July to November 2001, to investigate the biology and the involvement of each anopheline species in malaria transmission. Mosquitoes were captured when landing on human volunteers and by pyrethrum spray catches. Twelve anopheline species were captured. Four species amounted to 97% of human-bait sampling: Anopheles gambiae molecular form S, An. arabiensis, An. funestus, and An. nili s.s. All An. gambiae and An. nili females were fed on human, whereas the anthropophilic rate was 94.5% for An. funestus and 88.9% for An. arabiensis. Plasmodium falciparum was the only malaria parasite found, and infecting only An. gambiae, An. arabiensis, An. funestus, and An. nili. The circumsporozoite rate was 4.5% for An. gambiae, 1.6% for An. arabiensis, 3.9% for An. funestus, and 2.1% for An. nili. During the period of study, the entomological inoculation rate was estimated to 264 infected bites. An. gambiae, An. arabiensis, An. funestus, and An. nili were responsible respectively of 56, 3, 20, and 21% of malaria transmission. This study shows for the first time the implication of An. nili in malaria transmission in this area and the complexity of the malaria vectorial system that should be taken into account for any malaria control strategy.

Landscape Ecology of Sylvatic Chikungunya Virus and Mosquito Vectors in Southeastern Senegal

The risk of human infection with sylvatic chikungunya (CHIKV) virus was assessed in a focus of sylvatic arbovirus circulation in Senegal by investigating distribution and abundance of anthropophilic Aedes mosquitoes, as well as the abundance and distribution of CHIKV in these mosquitoes. A 1650 km2 area was classified into five land cover classes: forest, barren, savanna, agriculture and village. A total of 39,799 mosquitoes was sampled from all classes using human landing collections between June 2009 and January 2010. Mosquito diversity was extremely high, and overall vector abundance peaked at the start of the rainy season. CHIKV was detected in 42 mosquito pools. Our data suggest that Aedes furcifer, which occurred abundantly in all land cover classes and landed frequently on humans in villages outside of houses, is probably the major bridge vector responsible for the spillover of sylvatic CHIKV to humans.

Rift Valley Fever Outbreak with East-Central African Virus Lineage in Mauritania, 2003

Phylogenetic studies demonstrated that outbreak strains belonged to the East-Central African lineage.

Aspects of Bioecology of Two Rift Valley Fever Virus Vectors in Senegal (West Africa): Aedes vexans and Culex poicilipes (Diptera: Culicidae)

Abstract The dispersal, population dynamics, and age structure of two Rift Valley Fever Virus (Phlebovirus: Bunyaviridae) (RVFV) vectors, Aedes vexans Meigen and Culex poicilipes Theobald, were investigated in northern Senegal. The main objective was to investigate possible factors that mediate RVFV emergence and propagation at a site where humans and livestock live in proximity to temporary surface pools. In mark–release–recapture studies, recapture rates of 0.18% (156/85,500) and 3.46% (201/5,800) were obtained for Ae. vexans and Cx. poicilipes, respectively. The number of mosquitoes recaptured decreased with increasing distance from the release point and over time. The estimated daily survival rate for released females ranged from 91 to 96% for Ae. vexans and 70–79% for Cx. poicilipes. The maximum time after release when marked mosquitoes were collected was 23 and 12 d for Ae. vexans and Cx. poicilipes, respectively. The maximum distances from the release point that marked females were recaptured was 620 and 550 m for Ae vexans and Cx. poicilipes, respectively. Rainfall periodicity was a key factor controlling Ae. vexans population abundance. In contrast, rainfall had no discernible effect on the fluctuation of Cx. poicilipes numbers. The involvement of these two species in the transmission of RVFV is discussed with respect to their longevity and daily survival rate.

First attempt to validate the gSG6-P1 salivary peptide as an immuno-epidemiological tool for evaluating human exposure to Anopheles funestus bites.

Objective  The development of a biomarker of exposure based on the evaluation of the human antibody response specific to Anopheles salivary proteins seems promising in improving malaria control. The IgG response specific to the gSG6‐P1 peptide has already been validated as a biomarker of An. gambiae exposure. This study represents a first attempt to validate the gSG6‐P1 peptide as an epidemiological tool evaluating exposure to An. funestus bites, the second main malaria vector in sub‐Saharan Africa.

Comparisons of Human-Landing Catches and Odor-Baited Entry Traps for Sampling Malaria Vectors in Senegal

Abstract A comparative study of human-landing catches (HLCs) and odor-baited entry traps (OBETs) for sampling malaria vectors was conducted in two different bioclimatic areas of Senegal, the Sahelian and Sudano-Guinean phytogeographic zones, from September to December 2002. Mosquitoes were collected by the two methods both indoors and outdoors. The reliability of OBET samples was tested by comparing the two methods. Overall, HLC was more effective indoors and for surveying the anopheline fauna. Both methods were effective in sampling the four known malaria vectors in Senegal [Anopheles gambiae s.s., An. arabiensis Patton, An. funestus Giles, and An. nili (Theobald)], and mosquito age structures and infectivity rates did not differ between methods.

Population structure of the malaria vector Anopheles funestus in Senegal based on microsatellite and cytogenetic data

The study of chromosomal inversions distribution within natural Anopheles funestus populations from West Africa revealed high levels of genetic structuring. In Burkina Faso, this was interpreted as evidence for incipient speciation, and two chromosomal forms were described, namely ‘Folonzo’ and ‘Kiribina’. Assignation of field collected specimens to one chromosomal form depends upon application of an algorithm based on chromosomal inversions. We assessed relevance and applicability of this algorithm on An. funestus populations from Senegal, where both forms occur. Furthermore, we estimated the level of genetic differentiation between populations using microsatellite loci spread over the whole genome. Significant genetic differentiation was revealed between geographical populations of An. funestus, and the pattern observed suggested isolation by distance. Chromosomal heterogeneity was not detected by microsatellite markers. Thus, although incipient speciation could not be ruled out by our data, our results suggest that differential environmental selection pressure acting on inversions should be considered a major factor in shaping their distribution in wild An. funestus populations.