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      Multiple Insecticide Resistance in Anopheles gambiae s.l. Populations from Burkina Faso, West Africa

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          Abstract

          Malaria control programs are being jeopardized by the spread of insecticide resistance in mosquito vector populations. The situation in Burkina Faso is emblematic with Anopheles gambiae populations showing high levels of resistance to most available compounds. Although the frequency of insecticide target-site mutations including knockdown resistance (kdr) and insensitive acetylcholinesterase ( Ace-1 R) alleles has been regularly monitored in the area, it is not known whether detoxifying enzymes contribute to the diversity of resistance phenotypes observed in the field. Here, we propose an update on the phenotypic diversity of insecticide resistance in An. gambiae populations sampled from 10 sites in Burkina Faso in 2010. Susceptibility to deltamethrin, permethrin, DDT, bendiocarb and fenithrotion was assessed. Test specimens (N = 30 per locality) were identified to species and molecular form and their genotype at the kdr and Ace-1 loci was determined. Detoxifying enzymes activities including non-specific esterases (NSEs), oxydases (cytochrome P450) and Glutathione S-Transferases (GSTs) were measured on single mosquitoes (N = 50) from each test locality and compared with the An. gambiae Kisumu susceptible reference strain. In all sites, mosquitoes demonstrated multiple resistance phenotypes, showing reduced mortality to several insecticidal compounds at the same time, although with considerable site-to-site variation. Both the kdr 1014L and Ace-1 R 119S resistant alleles were detected in the M and the S forms of An. gambiae, and were found together in specimens of the S form. Variation in detoxifying enzyme activities was observed within and between vector populations. Elevated levels of NSEs and GSTs were widespread, suggesting multiple resistance mechanisms segregate within An. gambiae populations from this country. By documenting the extent and diversity of insecticide resistance phenotypes and the putative combination of their underlying mechanisms in An. gambiae mosquitoes, our work prompts for new alternative strategies to be urgently developed for the control of major malaria vectors in Burkina Faso.

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          The molecular basis of insecticide resistance in mosquitoes.

          Insecticide resistance is an inherited characteristic involving changes in one or more insect gene. The molecular basis of these changes are only now being fully determined, aided by the availability of the Drosophila melanogaster and Anopheles gambiae genome sequences. This paper reviews what is currently known about insecticide resistance conferred by metabolic or target site changes in mosquitoes.
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            Insecticide-treated bed nets and curtains for preventing malaria.

            C Lengeler (2004)
            Malaria is an important cause of illness and death in many parts of the world, especially in sub-Saharan Africa. There has been a renewed emphasis on preventive measures at community and individual levels. Insecticide-treated nets (ITNs) are the most prominent malaria preventive measure for large-scale deployment in highly endemic areas. To assess the impact of insecticide-treated bed nets or curtains on mortality, malarial illness (life-threatening and mild), malaria parasitaemia, anaemia, and spleen rates. I searched the Cochrane Infectious Diseases Group trials register (January 2003), CENTRAL (The Cochrane Library, Issue 1, 2003), MEDLINE (1966 to October 2003), EMBASE (1974 to November 2002), LILACS (1982 to January 2003), and reference lists of reviews, books, and trials. I handsearched journals, contacted researchers, funding agencies, and net and insecticide manufacturers. Individual and cluster randomized controlled trials of insecticide-treated bed nets or curtains compared to nets without insecticide or no nets. Trials including only pregnant women were excluded. The reviewer and two independent assessors reviewed trials for inclusion. The reviewer assessed trial methodological quality and extracted and analysed data. Fourteen cluster randomized and eight individually randomized controlled trials met the inclusion criteria. Five trials measured child mortality: ITNs provided 17% protective efficacy (PE) compared to no nets (relative rate 0.83, 95% confidence interval (CI) 0.76 to 0.90), and 23% PE compared to untreated nets (relative rate 0.77, 95% CI 0.63 to 0.95). About 5.5 lives (95% CI 3.39 to 7.67) can be saved each year for every 1000 children protected with ITNs. In areas with stable malaria, ITNs reduced the incidence of uncomplicated malarial episodes in areas of stable malaria by 50% compared to no nets, and 39% compared to untreated nets; and in areas of unstable malaria: by 62% for compared to no nets and 43% compared to untreated nets for Plasmodium falciparum episodes, and by 52% compared to no nets and 11% compared to untreated nets for P. vivax episodes. When compared to no nets and in areas of stable malaria, ITNs also had an impact on severe malaria (45% PE, 95% CI 20 to 63), parasite prevalence (13% PE), high parasitaemia (29% PE), splenomegaly (30% PE), and their use improved the average haemoglobin level in children by 1.7% packed cell volume. ITNs are highly effective in reducing childhood mortality and morbidity from malaria. Widespread access to ITNs is currently being advocated by Roll Back Malaria, but universal deployment will require major financial, technical, and operational inputs.
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              Identification of a point mutation in the voltage-gated sodium channel gene of Kenyan Anopheles gambiae associated with resistance to DDT and pyrethroids.

              A field trial of permethrin-impregnated bednets and curtains was initiated in Western Kenya in 1990, and a strain of Anopheles gambiae showing reduced susceptibility to permethrin was colonized from this site in 1992. A leucine-phenylalanine substitution at position 1014 of the voltage-gated sodium channel is associated with resistance to permethrin and DDT in many insect species, including Anopheles gambiae from West Africa. We cloned and sequenced a partial sodium channel cDNA from the Kenyan permethrin-resistant strain and we identified an alternative substitution (leucine to serine) at the same position, which is linked to the inheritance of permethrin resistance in the F(2) progeny of genetic crosses between susceptible and resistant individuals. The diagnostic polymerase chain reaction (PCR) developed by Martinez-Torres et al. [(1998) Insect Mol Biol 7: 179-184] to detect kdr alleles in field populations of An. gambiae will not detect the Kenyan allele and hence reliance on this assay may lead to an underestimate of the prevalence of pyrethroid resistance in this species. We adapted the diagnostic PCR to detect the leucine-serine mutation and with this diagnostic we were able to demonstrate that this kdr allele was present in individuals collected from the Kenyan trial site in 1986, prior to the introduction of pyrethroid-impregnated bednets. The An. gambiae sodium channel was physically mapped to chromosome 2L, division 20C. This position corresponds to the location of a major quantitative trait locus determining resistance to permethrin in the Kenyan strain of An. gambiae.
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                Author and article information

                Contributors
                Role: Editor
                Journal
                PLoS One
                PLoS ONE
                plos
                plosone
                PLoS ONE
                Public Library of Science (San Francisco, USA )
                1932-6203
                2012
                26 November 2012
                : 7
                : 11
                : e48412
                Affiliations
                [1 ]Institut de Recherche en Sciences de la Santé/Centre Muraz, Bobo-Dioulasso, Burkina Faso, West Africa
                [2 ]Maladies Infectieuses et Vecteurs: Ecologie, Génétique, Evolution et Contrôle, Institut de Recherche pour le Développement, Montpellier, France
                [3 ]Centre de Recherche Entomologique de Cotonou, Cotonou, Bénin
                [4 ]Centre de Coopération Internationale en Recherche Agronomique pour le Développement, Montpellier, France
                University of Crete, Greece
                Author notes

                Competing Interests: The authors have declared that no competing interests exist.

                Conceived and designed the experiments: RKD FS TB TM MN. Wrote the paper: MN FS. Performed the experiments under supervision of FS and RKD: MN. Analyzed the data under supervision of FS and RKD: MN. Participated to the sample collections: MN. Critically revised the paper: TB AD TM RKD. Institutional authority who facilitated the implementation of the work: JBO.

                Article
                PONE-D-12-14587
                10.1371/journal.pone.0048412
                3506617
                23189131
                c88a4072-a3ea-493a-acd1-e5395f1cf60e
                Copyright @ 2012

                This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

                History
                : 19 May 2012
                : 25 September 2012
                Page count
                Pages: 10
                Funding
                The study was supported by Corus project ref. 6015 and the IRD through the JEAI-RISA program. Corus project is a program of french research agency funding research in Southern universities and research institutes. MN was supported by a PhD scholarship from the IRD/DSF. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.
                Categories
                Research Article
                Agriculture
                Agrochemicals
                Pesticides
                Pest Control
                Pesticides
                Biology
                Microbiology
                Vector Biology
                Mosquitoes
                Medicine
                Infectious Diseases
                Parasitic Diseases
                Malaria
                Tropical Diseases (Non-Neglected)
                Malaria
                Vectors and Hosts
                Mosquitoes

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                Uncategorized

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