32 research outputs found

    Insecticide resistance status in Anopheles gambiae in southern Benin

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    BACKGROUND: The emergence of pyrethroid resistance in Anopheles gambiae has become a serious concern to the future success of malaria control. In Benin, the National Malaria Control Programme has recently planned to scaling up long-lasting insecticidal nets (LLINs) and indoor residual spraying (IRS) for malaria prevention. It is, therefore, crucial to monitor the level and type of insecticide resistance in An. gambiae, particularly in southern Benin where reduced efficacy of insecticide-treated nets (ITNs) and IRS has previously been reported. METHODS: The protocol was based on mosquito collection during both dry and rainy seasons across forty districts selected in southern Benin. Bioassay were performed on adults collected from the field to assess the susceptibility of malaria vectors to insecticide-impregnated papers (permethrin 0.75%, delthamethrin 0.05%, DDT 4%, and bendiocarb 0.1%) following WHOPES guidelines. The species within An. gambiae complex, molecular form and presence of kdr and ace-1 mutations were determined by PCR. RESULTS: Strong resistance to permethrin and DDT was found in An. gambiae populations from southern Benin, except in Aglangandan where mosquitoes were fully susceptible (mortality 100%) to all insecticides tested. PCR showed the presence of two sub-species of An. gambiae, namely An. gambiae s.s, and Anopheles melas, with a predominance for An. gambiae s.s (98%). The molecular M form of An. gambiae was predominant in southern Benin (97%). The kdr mutation was detected in all districts at various frequency (1% to 95%) whereas the Ace-1 mutation was found at a very low frequency (<or= 5%). CONCLUSION: This study showed a widespread resistance to permethrin in An. gambiae populations from southern Benin, with a significant increase of kdr frequency compared to what was observed previously in Benin. The low frequency of Ace-1 recorded in all populations is encouraging for the use of bendiocarb as an alternative insecticide to pyrethroids for IRS in Benin

    Resistance to DDT and Pyrethroids and Increased kdr Mutation Frequency in An. gambiae after the Implementation of Permethrin-Treated Nets in Senegal

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    Introduction: The aim of this study was to evaluate the susceptibility to insecticides of An. gambiae mosquitoes sampled in Dielmo (Senegal), in 2010, 2 years after the implementation of Long Lasting Insecticide-treated Nets (LLINs) and to report the evolution of kdr mutation frequency from 2006 to 2010. Methods: WHO bioassay susceptibility tests to 6 insecticides were performed on adults F0, issuing from immature stages of An. gambiae s.l., sampled in August 2010. Species and molecular forms as well as the presence of L1014F and L1014S kd

    Cotton pest management practices and the selection of pyrethroid resistance in Anopheles gambiae population in Northern Benin

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    <p>Abstract</p> <p>Background</p> <p>Pyrethroid insecticides, carbamate and organophosphate are the classes of insecticides commonly used in agriculture for crop protection in Benin. Pyrethroids remain the only class of insecticides recommended by the WHO for impregnation of bed nets. Unfortunately, the high level of pyrethroid resistance in <it>Anopheles gambiae </it>s.l., threatens to undermine the success of pyrethroid treated nets. This study focuses on the investigation of agricultural practices in cotton growing areas, and their direct impact on larval populations of <it>An. gambiae </it>in surrounding breeding sites.</p> <p>Methods</p> <p>The protocol was based on the collection of agro-sociological data where farmers were subjected to semi-structured questionnaires based on the strategies used for crop protection. This was complemented by bioassay tests to assess the susceptibility of malaria vectors to various insecticides. Molecular analysis was performed to characterize the resistance genes and the molecular forms of <it>An. gambiae</it>. Insecticide residues in soil samples from breeding sites were investigated to determine major factors that can inhibit the normal growth of mosquito larvae by exposing susceptible and resistant laboratory strains.</p> <p>Results</p> <p>There is a common use by local farmers of mineral fertilizer NPK at 200 kg/ha and urea at 50 kg/hectare following insecticide treatments in both the Calendar Control Program (CCP) and the Targeted Intermittent Control Program (TICP). By contrast, no chemicals are involved in Biological Program (BP) where farmers use organic and natural fertilizers which include animal excreta.</p> <p>Susceptibility test results confirmed a high resistance to DDT. Mean mortality of <it>An. gambiae </it>collected from the farms practicing CCP, TICP and BP methods were 33%, 42% and 65% respectively. <it>An. gambiae </it>populations from areas using the CCP and TICP programs showed resistance to permethrin with mortality of 50% and 58% respectively. By contrast, bioassay test results of <it>An. gambiae </it>from BP areas gave a high level of susceptibility to permethrin with an average mortality of 94%.</p> <p>Molecular analysis identified <it>An. gambiae </it>s.s, and <it>An. arabiensis </it>with a high predominance of <it>An. gambiae s.s </it>(90%). The two molecular forms, M and S, were also determined with a high frequency of the S form (96%).</p> <p>The <it>Kdr </it>gene seemed the main target- site resistance mechanism detected in CCP, TICP, and BP areas at the rates ranging from 32 to 78%. The frequency of <it>ace-1R </it>gene was very low (< 0.1).</p> <p>The presence of inhibiting factors in soil samples under insecticide treatments were found and affected negatively in delaying the development of <it>An. gambiae </it>larval populations.</p> <p>Conclusions</p> <p>This research shows that <it>Kdr </it>has spread widely in <it>An. gambiae</it>, mainly in CCP and TICP areas where pyrethroids are extensively used. To reduce the negative impact of pesticides use in cotton crop protection, the application of BP-like programs, which do not appear to select for vector resistance would be useful. These results could serve as scientific evidence of the spread of resistance due to a massive agricultural use of insecticides and contribute to the management of pesticides usage on cotton crops hence reducing the selection pressure of insecticides on <it>An. gambiae </it>populations.</p

    Field-based evidence of fast and global increase of Plasmodium falciparum drug-resistance by DNA-microarrays and PCR/RFLP in Niger

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    <p>Abstract</p> <p>Background</p> <p>Over the last years, significant progress has been made in the comprehension of the molecular mechanism of malaria resistance to drugs. Together with <it>in vivo </it>tests, the molecular monitoring is now part of the survey strategy of the <it>Plasmodium </it>sensitivity. Currently, DNA-microarray analysis allows the simultaneous study of many single nucleotide polymorphisms (SNP) of <it>Plasmodium </it>isolates. In December 2005, the International Federation of the Red Cross distributed two million three hundred thousand long-lasting insecticide nets to pregnant women and mothers of under five years children in the whole Niger. Then, Niger adopted artemisinin-based combination therapy as first-line treatment.</p> <p>Methods</p> <p>Thirty four SNPs of <it>pfcrt, pfdhfr, pfdhps, pfmdr </it>and <it>pfATPase </it>were analysed by DNA-microarray and PCR/RFLP in two villages – Zindarou and Banizoumbou – with different durations of malaria transmission. The main objective of the study was to measure the dynamics <it>of Plasmodium falciparum </it>resistant strains and associated factors.</p> <p>Results</p> <p>This study shows a global and clear increase of the drug-resistance associated molecular markers frequencies during a relatively short-time period of four years. Markers associated with resistance to chloroquine and sulphonamids were more frequently found in the short transmission zone than in the long transmission one. The <it>pfcrt76T </it>mutation is significantly more present at Banizoumbou than Zindarou (38.3% vs 25.2%, p = 0.013).</p> <p>This work allowed the screening of several field strains for five SNPs of <it>PfATPase6 </it>gene. The <it>pfATPase6S769N</it>, candidate mutation of resistance to artemisinin was not found. However the <it>pfATPsaeA623E </it>mutation was found in 4.7% of samples.</p> <p>Conclusion</p> <p>A significant increase of several SNPs frequencies was highlighted over a four-year period. The polymorphism of five <it>PfATPase6 </it>gene SNPs was described. The global, large and fast increase of the molecular resistance is discussed in the context of current changes of health policy and malaria control in Niger.</p

    Impact of three years of large scale Indoor Residual Spraying (IRS) and Insecticide Treated Nets (ITNs) interventions on insecticide resistance in Anopheles gambiae s.l. in Benin

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    <p>Abstract</p> <p>Background</p> <p>In Benin, Indoor Residual Spraying (IRS) and long-lasting insecticidal nets (LLINs) are the cornerstones of malaria prevention. In the context of high resistance of <it>Anopheles gambiae </it>to pyrethroids, The National Malaria Control Program (NMCP) has undertaken a full coverage of IRS in a no-flood zone in the Oueme region, coupled with the distribution of LLINs in a flood zone. We assessed the impact of this campaign on phenotypic resistance, <it>kdr </it>(knock-down resistance) and <it>ace-1<sup>R </sup></it>(insensitive acetylcholinesterase) mutations.</p> <p>Methods</p> <p>Insecticides used for malaria vector control interventions were bendiocarb WP (0.4 g/m<sup>2</sup>) and deltamethrin (55 mg/m<sup>2</sup>), respectively for IRS and LLINs. Susceptibility status of <it>An. gambiae </it>was assessed using World Health Organization bioassay tests to DDT, permethrin, deltamethrin and bendiocarb in the Oueme region before intervention (2007) and after interventions in 2008 and 2010. <it>An. gambiae </it>specimens were screened for identification of species, molecular M and S forms and for the detection of the West African <it>kdr </it>(L1014F) as well as <it>ace-1<sup>R </sup></it>mutations using PCR techniques.</p> <p>Results</p> <p>The univariate logistic regression performed showed that <it>kdr </it>frequency has increased significantly during the three years in the intervention area and in the control area. Several factors (LLINs, IRS, mosquito coils, aerosols, use of pesticides for crop protection) could explain the selection of individual resistant <it>An. gambiae</it>. The <it>Kdr </it>resistance gene could not be the only mechanism of resistance observed in the Oueme region. The high susceptibility to bendiocarb is in agreement with a previous study conducted in Benin. However, the occurrence of <it>ace-1<sup>R </sup></it>heterozygous individuals even on sites far from IRS areas, suggests other factors may contribute to the selection of resistance other than those exerted by the vector control program.</p> <p>Conclusion</p> <p>The results of this study have confirmed that <it>An.gambiae </it>have maintained and developed the resistance to pyrethroids, but are still susceptible to bendiocarb. Our data clearly shows that selection of resistant individuals was caused by other insecticides than those used by the IRS and LLINs.</p

    Low and seasonal malaria transmission in the middle Senegal River basin: identification and characteristics of Anopheles vectors

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    <p>Abstract</p> <p>Background</p> <p>During the last decades two dams were constructed along the Senegal River. These intensified the practice of agriculture along the river valley basin. We conducted a study to assess malaria vector diversity, dynamics and malaria transmission in the area.</p> <p>Methods</p> <p>A cross-sectional entomological study was performed in September 2008 in 20 villages of the middle Senegal River valley to evaluate the variations of <it>Anopheles </it>density according to local environment. A longitudinal study was performed, from October 2008 to January 2010, in 5 selected villages, to study seasonal variations of malaria transmission.</p> <p>Results</p> <p>Among malaria vectors, 72.34% of specimens collected were <it>An. arabiensis</it>, 5.28% <it>An. gambiae </it>of the S molecular form, 3.26% M form, 12.90% <it>An. pharoensis</it>, 4.70% <it>An. ziemanni</it>, 1.48% <it>An. funestus </it>and 0.04% <it>An. wellcomei</it>. <it>Anopheles </it>density varied according to village location. It ranged from 0 to 21.4 <it>Anopheles</it>/room/day and was significantly correlated with the distance to the nearest ditch water but not to the river.</p> <p>Seasonal variations of <it>Anopheles </it>density and variety were observed with higher human biting rates during the rainy season (8.28 and 7.55 <it>Anopheles </it>bite/man/night in October 2008 and 2009 respectively). Transmission was low and limited to the rainy season (0.05 and 0.06 infected bite/man/night in October 2008 and 2009 respectively). During the rainy season, the endophagous rate was lower, the anthropophagic rate higher and L1014F kdr frequency higher.</p> <p>Conclusions</p> <p>Malaria vectors are present at low-moderate density in the middle Senegal River basin with <it>An. arabiensis </it>as the predominant species. Other potential vectors are <it>An. gambiae </it>M and S form and <it>An. funestus</it>. Nonetheless, malaria transmission was extremely low and seasonal.</p

    Dynamics of insecticide resistance in malaria vectors in Benin: first evidence of the presence of L1014S kdr mutation in Anopheles gambiae from West Africa

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    <p>Abstract</p> <p>Background</p> <p>Insecticide resistance monitoring is essential to help national programmers to implement more effective and sustainable malaria control strategies in endemic countries. This study reported the spatial and seasonal variations of insecticide resistance in malaria vectors in Benin, West Africa.</p> <p>Methods</p> <p><it>Anopheles gambiae s.l </it>populations were collected from October 2008 to June 2010 in four sites selected on the basis of different use of insecticides and environment. WHO susceptibility tests were carried out to detect resistance to DDT, fenitrothion, bendiocarb, permethrin and deltamethrin. The synergist piperonyl butoxide was used to assess the role of non-target site mechanisms in pyrethroid resistance. <it>Anopheles gambiae </it>mosquitoes were identified to species and to molecular M and S forms using PCR techniques. Molecular and biochemical assays were carried out to determine <it>kdr </it>and <it>Ace.1<sup>R </sup></it>allelic frequencies and activity of the detoxification enzymes.</p> <p>Results</p> <p>Throughout the surveys very high levels of mortality to bendiocarb and fenitrothion were observed in <it>An. gambiae s.l</it>. populations. However, high frequencies of resistance to DDT and pyrethroids were seen in both M and S form of <it>An. gambiae s.s</it>. and <it>Anopheles arabiensis</it>. PBO increased the toxicity of permethrin and restored almost full susceptibility to deltamethrin. <it>Anopheles gambiae s.l</it>. mosquitoes from Cotonou and Malanville showed higher oxidase activity compared to the Kisumu susceptible strain in 2009, whereas the esterase activity was higher in the mosquitoes from Bohicon in both 2008 and 2009. A high frequency of <it>1014F kdr </it>allele was initially showed in <it>An. gambiae </it>from Cotonou and Tori-Bossito whereas it increased in mosquitoes from Bohicon and Malanville during the second year. For the first time the <it>L1014S kdr </it>mutation was found in <it>An. arabiensis </it>in Benin. The <it>ace.1<sup>R </sup></it>mutation was almost absent <it>in An. gambiae s.l</it>.</p> <p>Conclusion</p> <p>Pyrethroid and DDT resistance is widespread in malaria vector in Benin and both metabolic and target site resistance are implicated. Resistance was not correlated with a change of malaria species and/or molecular forms. The <it>1014S kdr </it>allele was first identified in wild population of <it>An. arabiensis </it>hence confirming the expansion of pyrethroid resistance alleles in Africa.</p

    Fine scale spatial investigation of multiple insecticide resistance and underlying target-site and metabolic mechanisms in Anopheles gambiae in central Cîte d’Ivoire

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    Routine monitoring of occurrence, levels and mechanisms of insecticide resistance informs effective management strategies, and should be used to assess the effect of new tools on resistance. As part of a cluster randomised controlled trial evaluating a novel insecticide-based intervention in central CĂŽte d’Ivoire, we assessed resistance and its underlying mechanisms in Anopheles gambiae populations from a subset of trial villages. Resistance to multiple insecticides in An. gambiae s.s. and An. coluzzii was detected across villages, with dose–response assays demonstrating extremely high resistance intensity to the pyrethroid deltamethrin (> 1,500-fold), and mortality following exposure to pyrethroid-treated bednets was low (< 30% mortality in cone bioassays). The 1014F kdr mutation was almost fixed (≄ 90%) in all villages but the 1575Y kdr-amplifying mutation was relatively rare (< 15%). The carbamate and organophosphate resistance-associated Ace-1 G119S mutation was also detected at moderate frequencies (22–43%). Transcriptome analysis identified overexpression of P450 genes known to confer pyrethroid resistance (Cyp9K1, Cyp6P3, and Cyp6M2), and also a carboxylesterase (COEAE1F) as major candidates. Cyp6P3 expression was high but variable (up to 33-fold) and correlated positively with deltamethrin resistance intensity across villages (r2 = 0.78, P = 0.02). Tools and strategies to mitigate the extreme and multiple resistance provided by these mechanisms are required in this area to avoid future control failures
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