Species Identification and Resistance Status of Anopheles gambiae s.l. (Diptera: Culicidae) Mosquitoes in Guinea.

Insecticide resistance is one of the primary threats to the recent gains in malaria control. This is especially true in Guinea, where long-lasting insecticidal nets are currently the primary vector control intervention. To better inform the national malaria control program on the current status of insecticide resistance in Guinea, resistance bioassays were conducted, using Anopheles gambiae s.l. Giles, in three sites. Molecular analyses were also done on An. gambiae s.l. to determine the species and find whether the target-site mutations kdr and Ace1R were present. Susceptibility tests revealed resistance to DDT and pyrethroids, although mosquitoes were susceptible to deltamethrin in two of the three sites tested. Mosquitoes were susceptible to bendiocarb, except in Kissidougou, Guinea. The kdr-west mutation was widespread and the frequency was 60% or more in all sites. However, the Ace1R mutation was present in low levels. Insecticide susceptibility should continue to be monitored in Guinea to ensure insecticide-based vector control methods remain effective.

Insecticide resistance in the Anopheles gambiae complex in Benin: a nationwide survey.

Benin has embraced World Health Organization-recommended preventive strategies to control malaria. Its National Malaria Control Programme is implementing and/or coordinating various actions and conducting evaluation trials of mosquito control strategies. Mosquito control is based on the use of insecticide-treated nets and indoor residual spraying, but the efficacy of these strategies to control malaria vectors is endangered by insecticide resistance. Here, we present the results of a nationwide survey on the status of insecticide susceptibility and resistance in Anopheles gambiae s.l. (Diptera: Culicidae) carried out in Benin in 2006-2007 (i.e. before extensive vector control was undertaken). Overall, our study showed that the S molecular form of An. gambiae s.s. predominates and is widely distributed across the country, whereas the frequency of the M form shows a strong decline with increasing latitude. Susceptibility to DDT, permethrin, carbosulfan and chlorpyrifos-methyl was assessed; individual mosquitoes were identified for species and molecular forms, and genotyped for the kdr and ace-1 loci. Full susceptibility to chlorpyrifos-methyl was recorded and very few samples displayed resistance to carbosulfan. High resistance levels to permethrin were detected in most samples and almost all samples displayed resistance to DDT. The kdr-Leu-Phe mutation was present in all localities and in both molecular forms of An. gambiae s.s. Furthermore, the ace-1(R) mutation was predominant in the S form, but absent from the M form. By contrast, no target modification was observed in Anopheles arabiensis. Resistance in the An. gambiae S molecular form in this study seemed to be associated with agricultural practices. Our study showed important geographic variations which must be taken into account in the vector control strategies that will be applied in different regions of Benin. It also emphasizes the need to regularly monitor insecticide resistance across the country and to adapt measures to manage resistance.

[Will climate and demography have a major impact on malaria in sub-Saharan Africa in the next 20 years?]. / Le climat et la démographie peuvent-ils avoir un impact important sur le paludisme en afrique subsaharienne dans les 20 prochaines années?

The purpose of this review of the literature is to present factors possibly affecting the spread of malaria in sub-Saharan Africa over the next 20 years. Malaria is a vector-borne disease that depends on environmental and human constraints. The main environmental limitations involve susceptibility of the vector (mosquitoes of the Anopheles genus) and parasite (Plasmodium falciparum) to climate. Malaria is a stable, endemic disease over most of the African continent. Climatic change can only affect a few regions on the fringes of stable zones (e.g. altitude areas or Sahel) where malaria is an unstable, epidemic disease. Higher temperatures could induce a decrease of malaria transmission in regions of the Sahel or an increase in the highlands. The extent of these overall trends will depend on the unpredictable occurrence of major meteorological phenomenon as well as on human activities affecting the environment that could lead to dramatic but limited outbreaks in some locations. The most influential human factors could be runaway demographic growth and urban development. Estimations based on modeling studies indicate that urbanization will lead to a 53.5% drop in exposure to malaria by 2030. However this reduction could be less than expected because of adaptation of Anopheles gambiae and An. arabiensis, the main vectors of malaria in sub-Saharan Africa, to the urban environment as well as increasing vector resistance to insecticides. Another unforeseeable factor that could induce unexpected malaria epidemics is mass migration due to war or famine. Finally immunosuppressive illnesses (e.g. HIV and malnutrition) could alter individual susceptibility to malaria. Social constraints also include human activities that modify land use. In this regard land use (e.g. forest clearance and irrigation) is known to influence the burden of malaria that is itself dependent on local determinants of transmission. Overall the most important social constraint for the population will be access to malarial prevention and implementation action to control this scourge.

Is K-O Tab 1-2-3((R)) long lasting on non-polyester mosquito nets?

Many societies use locally sourced mosquito nets made from a variety of materials. For protecting against malaria these require regular re-treatment with insecticide. K-O Tab 1-2-3 is a 'dip-it-yourself' long-lasting formulation with time-limited interim recommendation from WHO for treatment of washed white and coloured polyester nets for up to 15 washes. To determine wash-resistance on different fabrics, nets made of polyester, polyethylene, cotton or nylon were treated with K-O Tab 1-2-3 and washed up to 20 times using standard WHO washing procedures. Efficacy was assessed using cone and cylinder bioassays and tunnel tests, and deltamethrin content using high-pressure liquid chromatography. Polyethylene and cotton nets treated with K-O Tab 1-2-3 and washed 20 times achieved the WHO threshold of >80% mortality in tunnel tests. Polyethylene matched the performance of polyester in all bioassays in contrast to cotton and nylon which produced low mortality and knock-down in cone and cylinder bioassays. After 20 washes 16.5% of the loading dose of deltamethrin remained on the polyester nets compared with 28.7% on polyethylene, 38.9% on cotton and 2.2% on nylon. Cotton nets retained a high concentration of insecticide but the relatively poor performance in terms of knock-down and mortality suggest most insecticide is bound within the cotton fibres rather than on the surface. K-O Tab 1-2-3 renders insecticide wash fast on polyethylene nets, less so on cotton and nylon. Nets made from polyethylene can be treated in the home to render the insecticide long lasting.

Le climat et la demographie peuvent-ils avoir un impact important sur le paludisme en Afrique subsaharienne dans les 20 prochaines annees?

Cette revue de la litterature presente les conditions d'evolution du paludisme en Afrique subsaharienne dans les 20 prochaines annees. Le paludisme est une maladie vectorielle limitee par des contraintes environnementales et humaines. Les limites environnementales sont essentiellement dictees par la sensibilite du vecteur (moustiques du genreAnopheles) et du parasite Plasmodium falciparum au climat. Le paludisme est stable; dit endemique; sur une grande superficie de l'Afrique. Seules les zones ou le paludisme est instable; dit epidemique; principalement les franges de la zone stable (Sahel; altitude) pourront etre influencees par les changements climatiques. L'accroissement de la temperature; notamment; pourrait induire une reduction du paludisme en zone sahelienne ou une augmentation en altitude. Ces tendances globales devraient etre modulees par des evenements meteorologiques exceptionnels conjugues aux activites humaines s'exercant sur l'environnement qui pourront entrainer localement des epidemies dramatiques de paludisme. Les contraintes humaines impliquent en particulier une demographie galopante et un developpement des villes. Des modelisateurs projettent que l'urbanisation va entrainer une reduction de l'exposition palustre de 53;5en 2030. Toutefois; l'adaptation au milieu urbain d'Anopheles gambiae et d'An. arabiensis; principaux vecteurs du paludisme en Afrique subsaharienne; ainsi que leur resistance croissante aux insecticides; pourraient influencer cette diminution. De maniere imprevisible; les mouvements massifs de population resultant de guerres ou de famines pourront aussi entrainer des epidemies palustres inattendues. Enfin les maladies immunosuppressives (HIV; malnutrition) pourraient alterer la susceptibilite des individus au paludisme. Les contraintes sociales impliquent l'activite anthropique qui modifie l'utilisation des sols. On sait que l'utilisation des terres (deforestation; irrigation)peut influencer le poids du paludisme qui lui-meme depend des determinants locaux de la transmission. In fine; la contrainte sociale la plus importante demeure l'acces des populations aux interventions pour prevenir et lutter contre le paludisme

Presence of insensitive acetylcholinesterase in wild populations of Culex pipiens quinquefasciatus from Benin.

Resistance to organophosphates and carbamates was investigated in three samples of Culex pipiens quinquefasciatus Say, from Cotonou and Parakou in the Republic of Benin. Each population of larvae was selected with propoxur to eliminate susceptible individuals and more easily analyse resistant mosquitoes if the resistance allele is present in the field sample. The mass-selection showed that there are resistant mosquitoes in natural populations of C. p. quinquefasciatus in Benin and that nowadays they are still at a low frequency. The result of the PCR-based assay revealed the presence of the G119S mutation in all the C. p. quinquefasciatus mosquitoes that survived exposure to mass-selection. This mutation is the same than reported in a large range of insects, including Anopheles gambiae. Thus, the G119S mutation is spreading in natural populations of C. p. quinquefasciatus in Africa. Therefore, good mosquito control strategies and agricultural use of insecticides are critical, and should be carefully applied to limit the spread of this resistance gene.

Loss of protection with insecticide-treated nets against pyrethroid-resistant Culex quinquefasciatus mosquitoes once nets become holed: an experimental hut study.

BACKGROUND: An important advantage of pyrethroid-treated nets over untreated nets is that once nets become worn or holed a pyrethroid treatment will normally restore protection. The capacity of pyrethroids to kill or irritate any mosquito that comes into contact with the net and prevent penetration of holes or feeding through the sides are the main reasons why treated nets continue to provide protection despite their condition deteriorating over time. Pyrethroid resistance is a growing problem among Anopheline and Culicine mosquitoes in many parts of Africa. When mosquitoes become resistant the capacity of treated nets to provide protection might be diminished, particularly when holed. An experimental hut trial against pyrethroid-resistant Culex quinquefasciatus was therefore undertaken in southern Benin using a series of intact and holed nets, both untreated and treated, to assess any loss of protection as nets deteriorate with use and time. RESULTS: There was loss of protection when untreated nets became holed; the proportion of mosquitoes blood feeding increased from 36.2% when nets were intact to between 59.7% and 68.5% when nets were holed to differing extents. The proportion of mosquitoes blood feeding when treated nets were intact was 29.4% which increased to 43.6-57.4% when nets were holed. The greater the number of holes the greater the loss of protection regardless of whether nets were untreated or treated. Mosquito mortality in huts with untreated nets was 12.9-13.6%; treatment induced mortality was less than 12%. The exiting rate of mosquitoes into the verandas was higher in huts with intact nets. CONCLUSION: As nets deteriorate with use and become increasingly holed the capacity of pyrethroid treatments to restore protection is greatly diminished against resistant Culex quinquefasciatus mosquitoes.

Characterization of insensitive acetylcholinesterase (ace-1R) in Anopheles gambiae (Diptera: Culicidae): resistance levels and dominance.

Characterization of insecticide resistance provides data on the evolutionary processes involved in the adaptation of insects to environmental changes. Studying the dominance status and resistance level represents a great interest, in terms of understanding resistance evolution in the field to eventually adapt vector control. Resistance and dominance levels conferred by the G119S mutation of acetylcholinesterase (ace-1R) of the mosquito Anopheles gambiae s.s. (Diptera: Culicidae) were studied for various insecticides belonging to different classes, using strains sharing the same genetic background. Our survey shows that the homozygote resistant strain AcerKis displayed a very high resistance level to various carbamates (range 3,000- to 5,000-fold) compared with that of various organophosphates (range 12- to 30-fold). Furthermore, the dominance status varied between semi-recessivity with fenitrothion and chlorpyrifos methyl insecticides to semidominance with temephos, carbosulfan, and propoxur. These results indicate that this resistance mechanism could spread rapidly in the field and then compromise the use of organophosphate and carbamate compounds in public health. This study underlines the necessity to monitor the ace-1R mutation in natural populations before planning and implementing malaria control programs based on the use of these insecticides.

Preliminary field testing of a long-lasting insecticide-treated hammock against Anopheles gambiae and Mansonia spp. (Diptera: Culicidae) in West Africa.

The efficacy of an experimental long-lasting insecticide-treated hammock (LLIH) with a long-lasting treated net used as a blanket and made of the same fabric (polyethylene) was tested in a concrete block experimental hut, against the malaria vector Anopheles gambiae s.l. and the arbovirus vectors and nuisance mosquitoes Mansonia africana (Theobald) and Mansonia uniformis (Theobald). The LLIH was treated with the pyrethroid insecticide permethrin. It was evaluated concurrently with ignited mosquito coils over 20 successive weeks. In total, 2,227 mosquitoes (130 An. gambiae and 2,097 Mansonia spp.) corresponding to 27.8 mosquitoes per trap-night were collected in the untreated hut (control). The repellent effect of both coils and LLIH significantly reduced the number of mosquitoes entering the huts (35- 60%). There was no significant difference between LLIH and mosquito coils in blood-feeding inhibition (93-97%) or in mortality (88-98%). The LLIH is more cost-effective and user-friendly than mosquito coils, which need to be replaced nightly to protect people sleeping indoors from mosquito bites. The effects of LLIH on exophagic vectors also need to be investigated because most people that sleep in hammocks are outdoors.

Chlorfenapyr: a pyrrole insecticide for the control of pyrethroid or DDT resistant Anopheles gambiae (Diptera: Culicidae) mosquitoes.

Owing to the development and spread of pyrethroid resistance in Anopheles gambiae in Africa there is an urgent need to develop alternative insecticides to supplement the pyrethroids. Chlorfenapyr is a pyrrole insecticide first commercialized for the control of agricultural pests and termites. Performance against An. gambiae bearing kdr (pyrethroid and DDT resistance) or Ace-1(R) insensitive acetylcholinesterase (organophosphate and carbamate resistance) mechanisms was studied using a variety of adult bioassay tests including a simulated-experimental hut system (tunnel tests) that allows uninhibited mosquito behaviour/insecticide interactions. Strains resistant to pyrethroids and organophosphates showed no cross resistance to chlorfenapyr. In cone bioassays on treated netting the mortality of adult mosquitoes showed an unexpected curvilinear response, with highest mortality occurring at intermediate dosages. Adults expressed irritability to chlorfenapyr at higher dosages, which might explain the dosage-mortality trend. Toxic activity of chlorfenapyr was slow compared to conventional neurotoxic insecticides and additional mortality occurred between 24h and 72 h. In tunnel tests, the dosage-mortality trend showed a more typical sigmoid response and most mortality occurred during the first 24h. Mosquito penetration through the holed, treated netting showed only limited inhibition and blood-feeding was not inhibited. Mortality rates in the kdr strain exposed to chlorfenapyr treated netting in tunnel tests were much higher than with permethrin treated netting over the same 100-500 mg/m(2) dosage range. Chlorfenapyr has potential for malaria control in treated-net or residual spraying applications in areas where mosquitoes are pyrethroid resistant. For treated-net applications chlorfenapyr might be combined with pyrethroid as a mixture to provide personal protection as well as to give control of resistant mosquitoes.

Multiple insecticide resistance mechanisms in Anopheles gambiae and Culex quinquefasciatus from Benin, West Africa.

Because free-insecticide treated net distribution is planned in Benin (West Africa) during the next few years, we investigated the type, frequency and distribution of insecticide resistance mechanisms in Anopheles gambiae and Culex quinquefasciatus mosquitoes in four localities selected on the basis of contrasting agricultural practices, use of insecticides and environment. Bioassays with WHO diagnostic test kits were carried out using pyrethroid, carbamate, organophosphate and organochlorine insecticides. An. gambiae mosquitoes were identified to species and to M or S molecular forms using PCR techniques. Molecular and biochemical assays were carried out to identify kdr and Ace.1 mutations in individual mosquitoes and to detect any increase in the activity of enzymes typically involved in insecticide metabolism (oxidase, esterase and glutathion-S-transférases). WHO diagnostic tests showed high frequency of resistance in An. gambiae and Cx. quinquefasciatus to permethrin and DDT in three areas. This was consistent with the presence of target site insensitivity due to kdr mutation and to increased metabolism through enzymatic activity. Kdr was expressed in both M and S forms. However, less than 1% of An. gambiae or Cx. quiqnuefasciatus showed the presence of the Ace.1(R) mutation. Carbamate/OP resistance was present at higher frequency in Culex than in An. gambiae. Dieldrin resistance was present in both species at all four localities. A higher frequency of pyrethroid-resistance was found in An. gambiae mosquitoes collected in urban areas compared to those collected in rice growing areas. The expansion of vegetable growing within urban areas probably contributed to selection pressure on mosquitoes. The detection of multiple resistance mechanisms in both An. gambiae and Cx. quinquefasciatus in Benin may represent a threat for the efficacy of ITNs and other forms of vector control such as indoor residual spraying in the future.

A biological test to quantify pyrethroid in impregnated nets.

First instar larvae of Aedes aegypti (L.) (Diptera: Culicidae) are very susceptible to deltamethrin, which kills all larvae at a very low dose (0.1 microg/l). Thus the sensitivity of this insect to detect that pyrethroid is in parts per billion, as obtained with most common chromatographic methods of analysis. Here we describe a biological test (BT) to quantify deltamethrin in long-lasting insecticidal nets (LLINs) by exposing first instar larvae of Ae. aegypti to serial extracted insecticide solutions from net samples. The deltamethrin concentration in the net was calculated at the doses killing 50% of larva, from the LC(50) of deltamethrin (6.5 x 10(-5) mg/l) and the dilution factor (DF(50)) of the extracted net solution. The pyrethroid quantification in LLINs after 0-25 washes with this BT was correlated with those obtained by direct chromatographic analysis (r(2) = 0.84). This BT did not require sophisticated equipment and could be extended to other molecules and materials. It appeared accurate, robust, cheap and well adapted to the national malaria programmes as the eggs of Ae. aegypti might be used for some months. This method was adapted to provide an easy to use kit test for the quality control of LLINs in the field.

[Pyrethroid and DDT resistance of Anopheles gambiae s.s. (Diptera: Culicidae) in five agricultural ecosystems from Côte-d'Ivoire]. / Situation de la résistance d'anopheles gambiae s.s. (Diptera : Culicidae) aux pyréthrinoïdes et au DDT dans cinq ecosystèmes agricoles de Côte-d'Ivoire.

The promotion of pyrethroid impregnated bednets among the populations is a major activity of the National Malaria Vector Control Programs in African tropical countries. However pyrethroid resistance in Anopheles gambiae, a major malaria vector, has been observed in several parts of Côte-d'Ivoire since 1993. As insecticides used in agriculture against pests are frequently considered as important factors responsible for resistance in malaria vectors, we have evaluated insecticide resistance of An. gambiae populations taking into account the main crops cultivated in Côte-d'Ivoire. Five areas were selected according to the use of insecticide either in agriculture or for domestic purposes: a urban area where vegetables and rice are main crops, a rural rice growing area, a rural area of coffee/cocoa production, a rural area of fruit farming and a rural area without any use of agricultural insecticide. Susceptibility tests were carried out using WHO diagnostic test kits. About 6500 females of An. gambiae were exposed to insecticide impregnated papers (permethrin 1%, deltamethrin 0.05%, DDT 4%) for one hour. Results confirmed that both mortality rates and knockdown time analysis were important to study the resistance data. By using mortality rates, populations of An. gambiae were found to be resistant to pyrethroids and DDT in four of the five areas. Resistance was the highest in urban area, lower in fruit and coffee/cocoa areas and at low level in rice growing area. An. gambiae from area without agricultural treatment was found susceptible to pyrethroids but slightly resistant to DDT A significant increase of knockdown time was observed in all areas with the 3 insecticides. These results agreed with previous studies showing that kdr mutation was the main resistance mechanism to pyrethroids in An. gambiae populations in Côte-d'Ivoire. They also agreed with knockdown time which is an early indicator of resistance development for the population in area without agricultural treatment. In this population the frequency of homozygous resistant individuals was probably too low to have a significant decrease of mortality rates to pyrethroids.

[Influence of social perceptions and practices on the use of bednets in the malaria control programme in Ivory Coast (West Africa)]. / Representations sociales et pratiques liées à l'utilisation des moustiquaires dans la lutte contre le paludisme en Côte d'Ivoire (Afrique de l'Ouest).

The National Malaria Programme in Ivory Coast has encountered difficulty in winning public acceptance of insecticide-treated bednets. We speculate that resistance to the use of bednets could be rooted in social perceptions, beliefs and practices in the communities. The purpose of this study was to identify sociocultural and environmental factors that could be used to support promotion strategies and acceptance of impregnated bednets in Ivory Coast. Survey findings confirmed that bednets were not in widespread use among the population (25%). The most widely used methods were burning mosquito coils (50%) and indoor spraying (31%). Use of impregnated bednets was low (6%). Most survey respondents (73%) indicated initial appreciation for the effectiveness of bednets in protecting against mosquitoes as a nuisance. However only 9% of respondents thought that impregnated bednets provided protection against malaria although they did not necessarily use them. Design was a determinant factor for the use, and even acceptance, of bednets. The population want rectangular, permanently impregnated bednets large enough to accommodate at least 2 persons. Cost was a major obstacle to wider use by the population. According to our data the best price for the population would be between 2000 and 2500 FCFA as compared to the current price of 3500 FCFA in Ivory Coast.

Representations sociales et pratiques liees a l'utilisation des moustiquaires dans la lutte contre le paludisme en Cote d'Ivoire (Afrique de l'Ouest)

Devant les difficultes que rencontre le Programme National de Lutte contre le Paludisme (PNLP) de Cote d'Ivoire; dans sa volonte d'amener les populations a adopter les moustiquaires impregnees d'insecticide; nous nous sommes demandes si les representations sociales; les attitudes et les pratiques des communautes liees a l'utilisation des moustiquaires n'expliquent pas cette contre-performance. Notre etude a eu pour but de documenter les facteurs socioculturels et environnementaux pour renforcer les strategies de promotion et de vulgarisation de la moustiquaire impregnee en Cote d'Ivoire. Les resultats obtenus montrent qu'au niveau des enquetes quantitatives; la moustiquaire en general est faiblement utilisee par les populations (25). Les moyens les plus utilises sont les serpentins fumigenes (50) et les bombes aerosols (31). La moustiquaire impregnee est tres faiblement utilisee (6). La moustiquaire est; de maniere generale; appreciee dans un premier temps pour son efficacite dans la protection contre les nuisances dues aux moustiques par 73des enquetes. Seulement 9de ces enquetes pensent que la moustiquaire impregnee sert a se proteger contre le paludisme; mais ne l'utilisent pas necessairement. L'organisation des unites de couchage; voire leur inadaptation; sont determinantes dans l'utilisation de la moustiquaire. La moustiquaire souhaitee par les populations est celle d'au moins 2 places; de forme rectangulaire; de couleur blanche; faite en tulle a mailles fines; transparente; impregnee d'insecticide et a impregnation definitive. Toutefois; son cout semble etre le handicap majeur a son adoption par les populations; pour qui le cout ideal de la moustiquaire a l'achat; se situe entre 2000 et 2500 FCFA contre les 3 500 Frs actuellement pratiques en Cote d'Ivoire

[Use of agricultural insecticides in Benin]. / Utilisation des insecticides agricoles au Bénin.

The use of insecticides in households and in agriculture has been incriminated in the emergence of insecticide resistance in insect vectors. For farming staff, the emergence of vector resistance is due to indoors spray of insecticides using aerosols and other low quality products in rural and urban settings against mosquitoes. On the other hand, public health specialists believe that the phenomenon of resistance could be due to massive use of insecticides in agriculture for field pests control. In Turkey, the implication of agricultural use of pesticides in the selection of vector resistance is clearly established. This study was framed to identify potential practices favouring the emergence of insecticide resistance in the Republic of Benin. Interviews and focus group discussions were organized with cotton, rice and vegetables farmers. The final aim of these surveys was to point out practices likely to favour the emergence of resistance. The research is conducted in 3 cotton fields, 2 rice fields and 2 vegetable plantations. After filling and signing concerned forms, farmers are subjected to quantitative and qualitative questionnaires to generate data on: insecticides being used, the various doses applied for pests eradication, the frequency of treatments, the cost of treatments (cost/hectare/year) the origin of insecticides, the place of purchase, safety precautions and related health hazards. The results of this study have shown that the use of insecticides in agriculture is a clear fact. During treatments, insecticide residues get in contact with mosquito breeding sites where they diffuse into water and exercise a selection pressure on larvae. This partially explains the high levels of resistance recorded in with strains of Anopheles gambiae collected in agricultural settings under insecticides pressure. Pyrethroids and more specifically deltamethrin and cyfluthrin are the insecticides mainly used in studied localities. Bedrooms of farmers are used as storage place for half-used and un-used insecticides containers. For a proper management of insecticides, cotton and vegetable plantations farmers receive assistance from the ministry of rural development. Because of the importance of cotton production in the Republic of Benin, trainings on management of insecticides in agricultural settings are frequently organised by the ministry of rural development and are opened to farmers and their family members (husbands, wives, children and relatives...). In the long run, the whole family learns and becomes very versant with the use of insecticides, spraying frequencies, spraying devices and spraying techniques.

The spread of the Leu-Phe kdr mutation through Anopheles gambiae complex in Burkina Faso: genetic introgression and de novo phenomena.

During extensive sampling in Burkina Faso and other African countries, the Leu-Phe mutation producing the kdr pyrethroid resistance phenotype was reported in both Anopheles gambiae ss and A. arabiensis. This mutation was widely distributed at high frequency in the molecular S form of A. gambiae while it has been observed at a very low frequency in both the molecular M form and A. arabiensis in Burkina Faso. While the mutation in the M form is inherited through an introgression from the S form, its occurrence is a new and independent mutation event in A. arabiensis. Three nucleotides in the upstream intron of the kdr mutation differentiated A. arabiensis from A. gambiae ss and these specific nucleotides were associated with kdr mutation in A. arabiensis. Ecological divergences which facilitated the spread of the kdr mutation within the complex of A. gambiae ss in West Africa, are discussed.

Efficacy of mosquito nets treated with insecticide mixtures or mosaics against insecticide resistant Anopheles gambiae and Culex quinquefasciatus (Diptera: Culicidae) in Côte d'Ivoire.

Only pyrethroid insecticides have so far been recommended for the treatment of mosquito nets for malaria control. Increasing resistance of malaria vectors to pyrethroids threatens to reduce the potency of this important method of vector control. Among the strategies proposed for resistance management is to use a pyrethroid and a non-pyrethroid insecticide in combination on the same mosquito net, either separately or as a mixture. Mixtures are particularly promising if there is potentiation between the two insecticides as this would make it possible to lower the dosage of each, as has been demonstrated under laboratory conditions for a mixture of bifenthrin (pyrethroid) and carbosulfan (carbamate). The effect of these types of treatment were compared in experimental huts on wild populations of Anopheles gambiae Giles and the nuisance mosquito Culex quinquefasciatus Say, both of which are multi-resistant. Four treatments were evaluated in experimental huts over six months: the recommended dosage of 50 mg m(-2) bifenthrin, 300 mg m(-2) carbosulfan, a mosaic of 300 mg m(-2) carbosulfan on the ceiling and 50 mg m(-2) bifenthrin on the sides, and a mixture of 6.25 mg m(-2) carbosulfan and 25 mg m(-2) bifenthrin. The mixture and mosaic treatments did not differ significantly in effectiveness from carbosulfan and bifenthrin alone against anophelines in terms of deterrency, induced exophily, blood feeding inhibition and overall mortality, but were more effective than in earlier tests with deltamethrin. These results are considered encouraging, as the combination of different classes of insecticides might be a potential tool for resistance management. The mixture might have an advantage in terms of lower cost and toxicity.

Molecular evidence of incipient speciation within Anopheles gambiae s.s. in West Africa.

We karyotyped and identified by polymerase chain reaction restriction fragment length polymorphism (PCR-RFLP) analysis Anopheles gambiae s.s. samples collected in several African countries. The data show the existence of two non-panmictic molecular forms, named S and M, whose distribution extended from forest to savannahs. Mosquitoes of the S and M forms are homosequential standard for chromosome-2 inversions in forest areas. In dry savannahs, S is characterized mainly by inversion polymorphisms typical of Savanna and Bamako chromosomal forms, while M shows chromosome-2 arrangements typical of Mopti and/or Savanna and/or Bissau, depending on its geographical origin. Chromosome-2 inversions therefore seem to be involved in ecotypic adaptation rather than in mate-recognition systems. Strong support for the reproductive isolation of S and M in Ivory Coast comes from the observation that the kdr allele is found at high frequencies in S specimens and not at all in chromosomal identical M specimens. However, the kdr allele does not segregate with molecular forms in Benin.

[Lagoonal and coastal malaria at Cotonou: entomological findings]. / Le paludisme côtier lagunaire à Cotonou: données entomologiques.

Nowadays, malaria control is planned according to the epidemiological context. Various aspects of malaria have been described in sub-Saharan Africa. We report here entomological data from the coastal area of Benin, West Africa, which has many lakes and lagoons. We carried out a longitudinal study in which we investigated the dynamics of populations of malaria vectors in various zones, the frequency of inoculation in these zones, the infestation rate of the Anopheles gambiae mosquitoes collected, the effect of urbanization on malaria transmission, the effects of inundation and of salinity at mosquito breeding sites. A total of 3, 342 identifications were made on a chromosomal basis. Two species of the Anopheles gambiae complex were detected in the coastal and lagoon areas of Benin: An. melas and An. gambiae ss. The density of the populations of these species was highly dependent on the level of urbanization. In traditional villages on the lagoons (such as Agbalilamè, Djegbadji and Kétonou), the density of An. melas (86. 2%) was much higher than that in more urbanized areas (such as Ladji and Abomey-Calavi) (4.9%). We checked for chromosome polymorphism. We detected a 2Rn1 inversion in An. melas, similar to the 2Rn inversion found in mosquitoes in Gambia and Guinea-Bissau. The frequency of the n1 inversion and the density of An. melas populations were correlated and both seemed to depend on a single factor, salinity. The epidemiological situation with respect to malaria was very heterogeneous in the lagoon area of Benin. In the city of Cotonou, transmission was seasonal, sporozoite indices and the frequency of inoculation were high, in contrast to what would normally be expected in an urban area. In communities built on the beach, the level of transmission was markedly lower: about 5 infected bites per person per year versus 29 infected bites per person in the center of the city. In the traditional fishing villages, a paradoxical situation was observed in which the mosquitoes were very aggressive towards humans (4,502 bites per person per year) but the frequency of transmission was low (d = 0. 27%, CS+ = 0.57%). This was largely due to the high density in this area of An. melas, a poor malaria vector. If traditional villages become more urbanized, more freshwater breeding sites are created and the An. gambiae population increases, leading to an increase in malaria transmission. This is the reason for the higher level of malaria transmission at Ladji and Abomey-Calavi (h = 47 infected bites per person per year) than at Agbailamè, Djegbadji and Kétonou (h = 12.1 infected bites per person per year)