[Lessons from the Malaria Vector Control Program Based on Indoors Residual Spraying with DDT or Dieldrin in the Pilot Zone of Bobo-Dioulasso: Failure or Success?] / Les Leçons du Programme de Lutte Contre les Vecteurs du Paludisme Par Aspersions Intradomiciliaires de DDT ou de Dieldrine dans la Zone Pilote de Bobo-Dioulasso: Échec ou Succès?

During five years, from 1953, a village scale indoors residual spraying (IRS) was done in the pilot zone of Bobo-Dioulasso, Burkina Faso, with DDT or dieldrin (DLN) or even HCH with a conceptually both entomological and parasitological evaluation [18].Compared to the control area, DDT induced an approximatively 95% and 67% reduction in the landing rate of Anopheles gambiae, respectively inside and outside human houses but due to its irritant action, DDT greatly increased their exophagic behaviour. However, DLN had no impact on the landing rate of An. gambiae either indoors or outdoors due to the already noticed resistance of this species to this insecticide. The sporozoitic index of An. gambiae was reduced by 96% in the DDT treated areas and by 70% in the DLN treated area.DDT reduced the landing rates of Anopheles funestus by 98% and 91%, inside and outside treated houses respectively. With DLN, these reductions were 98% and 97%, respectively. The sporozoitic index of An. funestus was reduced by 95% in areas treated with DDT.Thus, vector control has reduced malaria transmission due to the two main vectors, An. gambiae and An. funestus, by some 99.8% in DDT treated villages compared to control villages. DLN reduced transmission from An. funestus by 99.9%, but almost not from An. gambiae . Overall, the implementation of vector control based on indoor residual spraying with DDT or DLN reduced by 99.9% the transmission of human Plasmodium in the villages of the pilot zone and therefore the program can be considered as entomologically successful.In children aged 2-9 years (target group for endemicity indices) the splenic index was 84.3% (n = 979) in the control area and 44.4% (n = 8920) in the treated areas (difference -47.3%), the plasmodial prevalence was 60.6% (n = 946) in the control zone and 38.0% (n = 7242) in the treated zones (difference - 37%) but the relatively high level of plasmodic or splenic index in treated villages showed that transmission was maintained at such a level that the program could be considered as a "semi-failure".Besides, the gametocytic indices remained at the same levels (3.28%, n = 946 in the control zone and 3.04%, n = 7242 in the treated zones) indicating the maintenance of the "reservoir of parasites" and the remaining possibilities of transmission.Compared to the control area, the index of new contamination was significantly lower in infants 0-3 months and 4 to 6 months in DDT treated villages but not in infants 7 to 12 months demonstrating that the control vector had some efficacy in the prevention of plasmodial infection but "all newborns were infected within one year" demonstrating that P. falciparum transmission was not completely stopped.In spite of its striking drop, the transmission was not fully stopped, and the programme was considered as a "semi-failure" or even a "failure" and inducing a complete shift in malaria control policy from vector control to mass drug chemotherapy (with several drugs, chloroquine, primaquine, pyriméthamine etc) without complete stop of transmission either. In fact, such vector control operations by DDT may have different analysis; in one side they can be considered an entomological success but, in another side, the actual reduction of plasmodic and splenic indices was not enough to be considered as successful. It was clear that both vector and parasite must be implemented in an integrated programme taking care of insecticide and drug resistance. Nevertheless, such programme, even not as successful as expected, could be considered as encouraging and not "disappointing" as it was. Important lessons can be learned from such large-scale field trial in spite of several methodological and operational issues.

[The five pillars of family and individual protection against mosquitoes, vectors of pathogenic agents]. / Les cinq piliers de la protection familiale et personnelle de l'homme contre les moustiques vecteurs d'agents pathogènes.

Except for yellow fever and encephalitis B, effective vaccines remain unavailable against most infectious diseases, and prevention is based mainly upon vector control and chemoprophylaxis. The increasing resistance of, respectively, mosquitoes and parasites to insecticides and drugs is thus a matter of great concern. The best approach is to combine chemoprophylaxis and entomoprophylaxis, that is, to use several methods of vector control at different levels to reduce the risk of infection and illness from these infectious diseases.

Assessment of geraniol-incorporated polymers to control Aedes albopictus (Diptera: Culicidae).

Effective control of mosquito borne diseases has proven extremely difficult with both vector and pathogen remaining entrenched and expanding in many disease endemic areas. When lacking an effective vaccine, vector control methods targeting both larval habitats and adult mosquito populations remain the primary strategy for reducing risk. Aedes albopictus from Thailand was used as a reference baseline for evaluation of natural insecticides incorporated in polymer disks and pellets and tested both in laboratory and field conditions. In laboratory and field tests, the highest larval mortality was obtained with disks or pellets containing IKHC (Insect Killer Highly Concentrate) from Fulltec AG Company. This product is reputed to contain geraniol as an active ingredient. With pellets, high mortality of Ae. albopictus larvae (92%) was observed in presence of 1 g of pellets per 500 ml of water at day 1st, and the mortality was 100% at day 1st for larvae in presence of 5 or 10 g of pellets. Fulltec AG Company has not accepted to give us the exact composition of their IKHC product. Therefore, we cannot recommend it, but the principle of using monoterpenes like geraniol, incorporated into polymer disks or pellets as natural larvicide needs more attention as it could be considered as a powerful alternative in mosquito vector control.

Entomological survey and report of a knockdown resistance mutation in the malaria vector Anopheles gambiae from the Republic of Guinea.

The purpose of our entomological survey was to estimate mosquito biodiversity, infectivity rates and insecticide resistance levels in Anopheles species in four study sites in a mining area with high malaria transmission in southeastern Guinea. Anopheles gambiae s.l. (77%) was the most common Anopheles collected followed by An. funestus (20%). The specimens of the An. gambiae complex were predominantly An. gambiae S form (97.6%) with 1.4% of An. gambiae M form found in Kérouané only, and 1% of An. arabiensis which was present in all four study sites. Anopheles gambiae S form and An. funestus were found to be infected with Plasmodium falciparum, with infectivity rates of 4.1% and 4.4% and inoculation rates of 0.60 and 0.19 infected bite/person/night, respectively. In addition, a high level (79%) of the knockdown resistance (kdr) L1014F mutation was reported in the populations of An. gambiae S form. The high malaria transmission that occurs in the prospected area of Guinea requires a long-term vector control programme. However, such a control programme will have to consider the presence of the kdr gene at a surprisingly high level within the dominant vector, which could reduce the expected impact of vector control.

Review on global co-transmission of human Plasmodium species and Wuchereria bancrofti by Anopheles mosquitoes.

Malaria and lymphatic filariasis are two of the most common mosquito-borne parasitic diseases worldwide which can occur as concomitant human infections while also sharing common mosquito vectors. This review presents the most recent available information on the co-transmission of human Plasmodium species and Wuchereria bancrofti by Anopheles mosquitoes. Important biological and epidemiological aspects are also described including the lifecycle of each parasite species and their specificities, the geographical biodiversity of each pathogen and their vectors where the parasites are co-endemic, and biological, environmental and climatic determinants influencing transmission. The co-transmission of each disease is illustrated from both a global perspective and a country level using Thailand as a study case. Different diagnostic methods are provided for the detection of the parasites in biological samples ranging from traditional to more recent molecular methods, including methodologies employing concomitant detection assays of W. bancrofti and Plasmodium spp. parasites. The relevant issues of combined malaria and Bancroftian filariasis control strategies are reviewed and discussed.

Bionomics, taxonomy, and distribution of the major malaria vector taxa of Anopheles subgenus Cellia in Southeast Asia: an updated review.

There is high diversity of Anopheles mosquitoes in Southeast Asia and the main vectors of malaria belong to complexes or groups of species that are difficult or impossible to distinguish due to overlapping morphological characteristics. Recent advances in molecular systematics have provided simple and reliable methods for unambiguous species identification. This review summarizes the latest information on the seven taxonomic groups that include principal malaria vectors in Southeast Asia, i.e. the Minimus, Fluviatilis, Culicifacies, Dirus, Leucosphyrus, and Sundaicus Complexes, and the Maculatus Group. Main issues still to be resolved are highlighted. The growing knowledge on malaria vectors in Southeast Asia has implications for vector control programs, the success of which is highly dependant on precise information about the biology and behavior of the vector species. Acquisition of this information, and consequently the application of appropriate, sustainable control measures, depends on our ability to accurately identify the specific vectors.

Review of the Minimus Complex of Anopheles, main malaria vector in Southeast Asia: from taxonomic issues to vector control strategies.

BACKGROUND: The Minimus Complex of Anopheles subgenus Cellia is composed of two sibling species, A and C, on the Southeast Asian mainland, and a third allopatric species E that occurs in the Ryukyu Archipelago (Japan), a malaria-free region. Anopheles minimus s.l. is considered to be one of the main malaria vector in the hilly forested regions of Southeast Asia. Despite a large number of studies over its range of distribution, it is difficult to have a global view of the ecological and bionomical characteristics of the individual species as different identification methods were used, generally without specific identification of the sibling species. OBJECTIVES: (1) To review the main malaria studies on An. minimus s.l.; (2) to discuss recently published data on the biology and ecology of each sibling species; and (3) to identify gaps in our understanding of the Minimus Complex. REVIEW RESULTS: Major biological and ecological trends are addressed, such as the high plasticity of trophic behaviour and the sympatry of species A and C over the Southeast Asian mainland. Despite the availability of rapid molecular identification methods, we still lack important information concerning the biological characteristics of each sibling species. These gaps must be filled in the future because An. minimus species A and C may exhibit different abilities to transmit malaria. CONCLUSION: We expect that entomological surveys will employ molecular methods to clearly identify these two species, and thus elucidate the biological characteristics of each species. As a consequence, current vector control strategies will be improved by targeting the most efficient vectors.

Restriction fragment length polymorphism method for the identification of major African and Asian malaria vectors within the Anopheles funestus and An. minimus groups.

The African Anopheles funestus and the Asian An. minimus groups are closely related and are probably considered distinct only because of their geographic separation. This study aimed at improving two identification methods based on polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) already developed for either group. Each PCR-RFLP, either on the internal transcribed spacer 2 (ITS2) for the An. minimus group, and domain 3 (D3) for the An. funestus group, was applied to the other group for the standardization of one identification method applicable on both continents. The ITS2 fragment digested by Bsi ZI showed the highest diagnostic power. This assay allowed the discrimination of at least 13 Anopheles species within the subgenus Cellia from two continents (Africa and Asia), among which are five major malaria vectors. Moreover, digestion of the D(3) with Msp I showed intragenomic variations within An. funestus populations. Two types of D3 copies (M and W) occurred in specimens from southern Africa. The populations from West-Central Africa presented only type W and East-Malagasy populations exhibited type M. Since An. funestus shows a great capacity of adaptation, these molecular variations, along with behavioral and ecologic ones, reinforce the hypothesis of a species complex that will need to be further investigated.

Pyrethroid and DDT cross-resistance in Aedes aegypti is correlated with novel mutations in the voltage-gated sodium channel gene.

Samples of the dengue vector mosquito Aedes aegypti (L.) (Diptera: Culicidae) were collected from 13 localities between 1995 and 1998. Two laboratory strains, Bora (French Polynesia) and AEAE, were both susceptible to DDT and permethrin; all other strains, except Larentuka (Indonesia) and Bouaké (Ivory Coast), contained individual fourth-instar larvae resistant to permethrin. Ten strains were subjected to a range of biochemical assays. Many strains had elevated carboxylesterase activity compared to the Bora strain; this was particularly high in the Indonesian strains Salatiga and Semarang, and in the Guyane strain (Cayenne). Monooxygenase levels were increased in the Salatiga and Paea (Polynesia) strains, and reduced in the two Thai strains (Mae Kaza, Mae Kud) and the Larentuka strain. Glutathione S-transferase activity was elevated in the Guyane strain. All other enzyme profiles were similar to the susceptible strain. The presence of both DDT and pyrethroid resistance in the Semarang, Belem (Brazil) and Long Hoa (Vietnam) strains suggested the presence of a knock-down resistant (kdr)-type resistance mechanism. Part of the S6 hydrophobic segment of domain II of the voltage-gated sodium channel gene was obtained by RT-PCR and sequenced from several insects from all 13 field strains. Four novel mutations were identified. Three strains contained identical amino acid substitutions at two positions, two strains shared a different substitution, and one strain was homozygous for a fourth alteration. The leucine to phenylalanine substitution that confers nerve insensitivity to pyrethroids in a range of other resistant insects was absent. Direct neurophysiological assays on individual larvae from three strains with these mutations demonstrated reduced nerve sensitivity to permethrin or lambda cyhalothrin inhibition compared to the susceptible strains.

SCAR markers and multiplex PCR-based identification of isomorphic species in the Anopheles dirus complex in Southeast Asia.

The Anopheles dirus Peyton & Harrison complex of mosquitoes (Diptera: Culicidae) comprises seven known species, including important malaria vectors in Southeast Asia. Specific identification of each species of the complex, which cannot be distinguished using morphological characters, is crucial for understanding vector ecology and implementing effective control measures. Derived from individual random amplified polymorphic DNA (RAPD) markers, sequence characterized amplified regions (SCAR) were analysed for the design of specific paired-primers. Combination of six SCAR primers resulted in the development of a simple, robust, single multiplex PCR able to identify three important malaria vectors among the four most common species (A, B, C, D) of the complex: species A from several Southeast Asian countries, species B from Perlis, Malaysia, and species C and D from Thailand.

[Molecular identification of sibling Anopheles species: example of the Anopheles minimus and Anopheles dirus complexes, major malarial vectors in Southeast Asia]. / Identification moléculaire d'espèces jumelles d'anophèles: exemple des complexes Anopheles minimus et Anopheles dirus vecteurs majeurs du paludisme en asie du Sud-Est.

Effective control of malaria vectors requires precise identification of species. This is especially important within complexes of species that cannot be distinguished based on morphological features. Two methods based on polymerase chain reaction (PCR) analysis have been developed to identify 2 species in the Anopheles minimus complex and 5 species of the Minimums group as well as 4 species of the Anopheles dirus complex. Association of oligonucleotide couples in the form of multiplex PCR has allowed development of two simple, reliable PCR techniques adapted to each one of these species complexes that comprise the major vectors of malaria in Southeast Asia. Specifically designed to meet the needs of entomologist working in the field for reliable, cost-effective tests, these techniques will facilitate assessment of the geographical distribution of each vector. These data will help to better target vector control measures.

A multiplex PCR-based method derived from random amplified polymorphic DNA (RAPD) markers for the identification of species of the Anopheles minimus group in Southeast Asia.

Effective control of Anopheles minimus s.l., an important malaria vector in Southeast Asia, is based on the accurate identification of species within An. minimus complex, which cannot be distinguished using morphological characters. Derived from individual random amplified polymorphic DNA markers, sequence characterized amplified regions were analysed for the design of species-specific paired-primers. Combination of these primers resulted in the development of a simple, robust multiplex PCR able to identify both species An. minimus A and C belonging to the complex, hybrids AC, and three sympatric and closely related species, An. aconitus, An. pampanai and An. varuna. Hybrids AC do not possess alleles of both parents but exhibit novel adaptive potentials resulting from recombination among parental genes leading to hybrizyme.

DDT house spraying and re-emerging malaria.

PIP: This article discusses the role of DDT in the re-emerging cases of malaria worldwide. It is noted that malaria is reappearing in urban areas and in countries that previously eradicated the disease, including the Amazon Basin, South and North Korea, Armenia, Azerbaijan, and Tajikistan. In addition, the frequency of imported malaria has also increased in industrial countries. Although many factors contribute to such a phenomenon, the strongest correlation is with decreasing numbers of houses sprayed with DDT. Early studies of DDT showed repellent, irritant, and toxic actions that worked against malaria vector mosquitoes. Sprayed on house walls, DDT exerted powerful control over indoor transmission of malaria. However, since the ban of DDT in the 1970s and the implementation of alternative malaria-control programs there has been a global outburst of malaria epidemics. In view of this, it is recommended that the global response to burgeoning malaria rates allow for DDT residual house spraying where it is known to be effective and necessary. Regulations and policies of industrialized countries and international agencies that block financial assistance to countries that use DDT for malaria control should be eliminated.^ieng

The kdr mutation occurs in the Mopti form of Anopheles gambiae s.s. through introgression.

Anopheles gambiae s.s. is a complex of sibling taxa characterized by various paracentric inversions. In west and central Africa, where several taxa are sympatric, a kdr mutation responsible for pyrethroid resistance has been described in only one (the S taxon), suggesting an absence of gene flow between them. Following a thorough sampling, we have found a kdr mutation in another taxon (M). To establish whether this mutation is the same event or not, the large intron upstream of the kdr mutation was sequenced to find polymorphic sites in susceptible/resistant and M/S mosquitoes. The low genetic diversity found in this DNA region indicates that a local genetic sweep has recently occurred. However, some polymorphic sites were found, and it is therefore concluded that the kdr mutation in the M taxon is not an independent mutation event, and is best explained by an introgression from the S taxon. These results are discussed within the context of possible gene flow between members of An. gambiae s.s. taxa, and with the possible spread of the kdr mutation in other closely related malaria vectors of the An. gambiae complex.

Modifications of pyrethroid effects associated with kdr mutation in Anopheles gambiae.

Effects of knockdown resistance (kdr) were investigated in three pyrethroid-resistant (RR) strains of the Afrotropical mosquito Anopheles gambiae Giles (Diptera: Culicidae): Kou from Burkina Faso, Tola and Yao from Côte d'Ivoire; compared with a standard susceptible (SS) strain from Kisumu, Kenya. The kdr factor was incompletely recessive, conferring 43-fold resistance ratio at LD50 level and 29-fold at LD95 level, as determined by topical application tests with Kou strain. When adult mosquitoes were exposed to 0.25% permethrin-impregnated papers, the 50% and 95% knockdown times (KdT) were 23 and 42 min for SS females, compared with 40 and 62 min for RS (F1 Kou x Kisumu) females. On 1% permethrin the KdT50 and KdT95 were 11 and 21 min for SS compared with 18 and 33 min for RS females. Following 1 h exposure to permethrin (0.25% or 1%), no significant knockdown of Kou RR females occurred within 24 h. Permethrin irritancy to An. gambiae was assessed by comparing 'time to first take-off' (TO) for females. The standard TO50 and TO95 values for Kisumu SS on untreated paper were 58 and 1044 s, respectively, vs. 3.7 and 16.5 s on 1% permethrin. For Kou RR females the comparable values were 27.3 s for TO50 and 294 s for TO95, with intermediate RS values of 10.1 s for TO50 and 71.9 s for TO95. Thus, TO values for RS were 2.7-4.4 times more than for SS, and those for RR were 7-18 times longer than for SS. Experiments with pyrethroid-impregnated nets were designed to induce hungry female mosquitoes to pass through holes cut in the netting. Laboratory 'tunnel tests' used a bait guinea-pig to attract mosquitoes through circular holes (5 x 1 cm) in a net screen. With untreated netting, 75-83% of laboratory-reared females passed through the holes overnight, 63-69% blood-fed successfully and 9-17% died, with no significant differences between SS and RR genotypes. When the netting was treated with permethrin 250mg ai/m2 the proportions that passed through the holes overnight were only 10% of SS vs. 40-46% of RR (Tola & Kou); mortality rates were 100% of SS compared with 59-82% of RR; bloodmeals were obtained by 9% of Kou RR and 17% of Tola RR, but none of the Kisumu SS females. When the net was treated with deltamethrin 25 mg ai/m2 the proportions of An. gambiae that went through the holes and blood-fed successfully were 3.9% of Kisumu SS and 3.5% of Yaokoffikro field population (94% R). Mortality rates were 97% of Kisumu SS vs. 47% of Yaokoffikro R. Evidently this deltamethrin treatment was sufficient to kill nearly all SS and half of the Yaokoffikro R An. gambiae population despite its high kdr frequency. Experimental huts at Yaokoffikro were used for overnight evaluation of bednets against An. gambiae females. The huts were sealed to prevent egress of mosquitoes released at 20.00 hours and collected at 05.00 hours. Each net was perforated with 225 square holes (2 x 2 cm). A man slept under the net as bait. With untreated nets, only 4-6% of mosquitoes died overnight and bloodmeals were taken by 17% of SS vs. 29% of Yaokoffikro R (P<0.05). Nets treated with permethrin 500 mg/m2 caused mortality rates of 95% Kisumu SS and 45% Yao R (P<0.001) and blood-feeding rates were reduced to 1.3% of SS vs. 8.1% of Yao R (P<0.05). Nets treated with deltamethrin 25 mg/m2 caused mortality rates of 91% Kisumu SS and 54% Yao R (P<0.001) and reduced blood-feeding rates to zero for SS vs. 2.5% for Yao R (P>0.05). (ABSTRACT TRUNCATED)

[Anopheline population genetics]. / Génétique des populations anophéliennes.

Population genetic studies of vectors are essential for (i) the determination of their taxonomic status and consequently the definition of their vectorial role in the transmission of pathogenic agents; (ii) the evaluation of the species genetic variability and the estimation of their capacities of adaptation to selection pressure; (iii) an estimation of gene flow among populations in order to evaluate their degree of isolation and gene circulation, especially resistance genes. Among the malaria vectors taken as examples on three continents, Africa, South-East Asia and Latin America, the large majority of the species showed an important polymorphism. The Gambiae Complex, which is by far the most studied one, includes at present 7 species with the recent description of An. quadriannulatus A and B from Ethiopia. An. gambiae s.s. includes itself 5 chromosomal forms. One of them, the Mopti form, should be considered as a species unto itself. For An. arabiensis, a strong differentiation has been observed among the populations from Senegal and the Indian Ocean Islands. The kdr mutation, which confers resistance to pyrethroid knockdown effect, has never been found either in the Mopti form, or An. arabiensis, indicating a restricted gene flow between these latter two and An. gambiae s.s. The speciation process of the Gambiae Complex seems to be a recent phenomenon due to environmental selection pressure. Species of the Funestus Group are distinguishable by morphological characters. The genetic study of An. funestus s.s. did not show the presence of a complex, in spite of high polymorphism and population structure. Anophelines from eastern areas present an important biodiversity. The Minimus Complex includes two species, A and C, which are widely distributed in South-East Asia. Species A is strongly endophilic, on the contrary species C is at once more exophilic and zoophilic. The latter species might have been selected by DDT indoor house spraying. After numerous taxonomic investigations, the Dirus Complex includes now 7 species. In Latin America, An. pseudopunctipennis clustered into three geographic populations which are under a speciation process. One covers North America and Guatemala, the other South America and Belize, whilst the last one is restricted to Grenada Island. On the contrary, An. darlingi showed little morphologic and genetic variability throughout the species geographic range suggesting the existence of a single species. The main objective of these studies is to implement a more selective approach of vector control programs in relation to the incriminated species, their bioecology and their role in malaria transmission. The improvement of efficiency and selectivity of vector control is becoming a major goal in order to make the best out of the available tools and control the impact of interventions on the environment.

Population structure of the primary malaria vector in South America, Anopheles darlingi, using isozyme, random amplified polymorphic DNA, internal transcribed spacer 2, and morphologic markers.

A genetic and morphologic survey of Anopheles darlingi populations collected from seven countries in Central and South America was performed to clarify the taxonomic status of this major malaria vector species in the Americas. Population genetics was based on three techniques including isozyme, random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR), and internal transcribed spacer 2 (ITS2) markers. The results of the isozyme analysis indicated moderate differences in the allele frequencies of three putative loci (glutamate oxalaoacetate transaminase-1, isocitrate dehydrogenase-1, and phosphoglucomutase) of the 31 analyzed. No fixed electromorphic differences separated the populations of An. darlingi, which showed little genetic divergence (Nei distances = 0.976-0.995). Fragments produced by RAPD-PCR demonstrated evidence of geographic partitioning and showed that all populations were separated by small genetic distances as measured with the 1 - S distance matrix. The ITS2 sequences for all samples were identical except for four individuals from Belize that differed by a three-base deletion (CCC). The morphologic study demonstrated that the Euclidean distances ranged from 0.02 to 0.14, with the highest value observed between populations from Belize and Bolivia. Based on these analyses, all the An. darlingi populations examined demonstrated a genetic similarity that is consistent with the existence of a single species and suggest that gene flow is occurring throughout the species' geographic range.

Pyrethroid cross resistance spectrum among populations of Anopheles gambiae s.s. from Côte d'Ivoire.

Field samples of Anopheles gambiae s.s. from Côte d'Ivoire were tested with 5 pyrethroids (cyfluthrin, lambda-cyhalothrin, alpha-cypermethrin, deltamethrin, permethrin), 1 pseudo-pyrethroid (etofenprox), and an organochlorine (DDT). With the use of World Health Organization diagnostic tests, 5 out of 6 samples were found cross-resistant to these insecticides. A strong decrease in knockdown effect and mortality was also observed when testing deltamethrin-impregnated nettings. With a polymerase chain reaction amplification of specific alleles diagnostic test, resistance was found associated with the presence of a kdr mutation. The strong correlation between kdr allelic frequency and resistance to DDT or etofenprox indicated that kdr was the main resistance factor for these 2 insecticides. On the contrary, a lower correlation was observed between kdr frequency and resistance to 4 of the 5 pyrethroids tested, suggesting that another mechanism was also involved, likely a metabolic detoxification. These results point out the necessity to monitor pyrethroid resistance and the presence of kdr before implementation of any impregnated bed-net programs for malaria control.

Current distribution of a pyrethroid resistance gene (kdr) in Anopheles gambiae complex from west Africa and further evidence for reproductive isolation of the Mopti form.

In the field, the kdr mutation, involved in pyrethroid resistance, has been found widely distributed in the Savanna form of Anopheles gambiae s.s., but never in wild populations of the Mopti form or An. arabiensis, even in areas where both occur in sympatry with resistant Savanna populations. Under laboratory conditions, Mopti and Savanna forms were fully able to interbreed and the kdr mutation was transmissible from one form to the other. Both forms appeared to be exposed to pyrethroid selection pressure in the field. The absence of the kdr mutation in the Mopti form and the total lack of Mopti-Savanna heterozygotes in field populations provides further evidence of a pre-copulatory barrier to gene flow between these two forms. Molecular markers, including kdr, are powerful tools for studying population genetics and circulation of resistance genes, and should be used through an integrated approach for a better understanding of the speciation process.