Bendiocarb, a potential alternative against pyrethroid resistant Anopheles gambiae in Benin, West Africa

<p>Abstract</p> <p>Background</p> <p><it>Anopheles gambiae</it>, the main malaria vector in Benin has developed high level of resistance to pyrethroid insecticides, which is a serious concern to the future use of long-lasting insecticidal nets (LLIN) and indoor residual spraying (IRS). In this context, one of the pathways available for malaria vector control would be to investigate alternative classes of insecticides with different mode of action than that of pyrethroids. The goal of this study was to evaluate under field conditions the efficacy of a carbamate (bendiocarb) and an organophosphate (fenitrothion) against pyrethroid-resistant <it>An. gambiae s.s</it>.</p> <p>Methods</p> <p>Wild populations and females from laboratory colonies of five days old <it>An. gambiae </it>were bio-assayed during this study. Two pyrethroids (deltamethrin and alphacypermethrin), an organophosphate (fenitrothion), a carbamate (bendiocarb) and a mixture of an organophosphate (chlorpyriphos + a pyrethroid deltamethrin) were compared in experimental huts as IRS treatments. Insecticides were applied in the huts using a hand-operated compression sprayer. The deterrency, exophily, blood feeding rate and mortality induced by these insecticides against <it>An. gambiae </it>were compared to the untreated control huts.</p> <p>Results</p> <p>Deltamethrin, alphacypermethrin and bendiocarb treatment significantly reduced mosquito entry into the huts (p < 0.05) compared to untreated huts. Blood feeding rates in huts treated with fenitrothion and the mixture chlorpyriphos/deltamethrin were reduced from 10.95% respectively to 3.7% and 4.47% three months after treatment and from 10.20% to 4.4% and 2.04% four months after treatment. Exophily rates in huts with deltamethrin, alphacypermethrin and the mixture chlorpyriphos/deltamethrin were significantly higher than in the huts with fenitrothion. Deltamethrin and alphacypermethrin had the lowest mortality rate while fenitrothion killed 100% of <it>An. gambiae </it>(in the first month) and 77.8% (in the fourth month). Bendiocarb and the mixture chlorpyriphos/deltamethrin mortality rates ranged from 97.9 to 100% the first month and 77.7-88% the third month respectively.</p> <p>Conclusion</p> <p>After four months, fenitrothion, bendiocarb and the mixture chlorpyriphos/deltamethrin performed effectively against pyrethroid-resistant <it>Anopheles</it>. These results showed that bendiocarb could be recommended as an effective insecticide for use in IRS operations in Benin, particularly as the mixture chlorpyriphos/deltamethrin does not have WHOPES authorization and complaints were mentioned by the sleepers about the safety and smell of fenitrothion.</p

Etude De L’efficacité Du Cashew Nut Shell Liquid (CNSL) De Anacardium Occidentale L. Extrait À Froid Sur Le Contrôle De Anopheles Gambiae S.L Résistant Aux Pyréthrinoïdes

In order to combat the vectors of malaria, it is necessary to know their behavior with regard to the insecticide products envisaged. The present study is aimed at the CNSL efficiency test extracted from the seed hull of Anacardium occidentale L. for the control of pyrethroid-resistant Anopheles gambiae s.l. Sensitivity tests were carried out on the larvae of stages 2, 3 and 4 of An. gambiae of the wild and Kis-kdr strains carrying the Kdr gene. The "Kisumu" strain devoid of resistance gene served as a control. The mortalities were read after 24 hours of exposure. LD50 and DL90 were determined using the log-probit method of dose determination corresponding to a proportion. The high LD50 determined is 0.0042 mg / ml and corresponds to the LD50 of the larvae of stage 2 of the wild strain. The low LD50 is determined on stage 2 larvae "Kisumu". The high DL90 is determined on stage 4 larvae of the wild strain. The low LD90 determined corresponds to stage 2 larvae "Kisumu". The comparison of the LD between the different strains shows a low sensitivity to CNSL of the larvae of resistant strains compared to the control strain, indicating the influence of the resistance factor kdr on the CNSL efficiency. However, the low-dose CNSL larvicidal activity on the larvae of the kdr-carrying strains provides an alternative advantage in the management of vector resistance to pyrethroid

Etude De L’efficacité De L’huile De Thevetia Neriifolia Pour Le Contrôle De Anopheles Gambiae S.L Résistant Aux Pyréthrinoïdes

Effective control of pyrethroid-resistant malaria vectors requires new alternative measures. The purpose of this study is to test the effectiveness of Thevetia oil extracted from seed kernels to control malaria transmission by An. gambiae s.l. Sensitivity tests were carried out on larvae of Stages 3 and 4 of An. gambiae s.s of wild-type and pyrethroid-resistant Kis-kdr. The susceptible reference strain "Kisumu" served as a control. Mortality was read 24 hours and 48 hours after exposure. LD50 and 90 for 24 hours and 48 hours were determined using the log-probit method of determining the dose corresponding to a proportion. The high LD50s determined in 24 hours and 48 hours correspond to doses that kill 50% of Stage 3 larvae in wild populations in 24 hours and 48 hours. Low LD50s refer to stage 4 "Kisumu" larvae. The strong DL90 in 24 hours and 48 hours correspond respectively to stage 3 larvae of the Kis-kdr and wild-type strains. The comparison of the LD between the different strains shows the influence of the resistance on the sensitivity of the larvae of the strains resistant to the oil tested. However, the larvicidal effect of Thevetia oil on the larvae of resistant strains may elicit formulations for alternative measures in vector resistance management to pyrethroids

Blood feeding behaviour comparison and contribution of Anopheles coluzzii and Anopheles gambiae, two sibling species living in sympatry, to malaria transmission in Alibori and Donga region, northern Benin, West Africa

Background: The main goal of this study was to assess the blood feeding behaviour and the contribution Anopheles coluzzii and Anopheles gambiae, 2 sibling species of An. gambiae sensu stricto. present and living in sympatry in 2 regions of northern Benin targeted for indoor residual spraying (IRS).Methods: The study was carried out in 6 districts of 2 regions of Benin (Alibori and Donga). Human landing catches (HLC) performed inside and outside of the households and pyrethrum spray captures (PSC) carried out in bedrooms were used to sample vector populations (An. gambiae and An. coluzzii). Collected mosquitoes were analysed to esti-mate the human biting rate indoors and outdoors, the circumsporozoite antigen positivity, and the anthropophagic index using ELISA methodology. Polymerase chain reaction was used to estimate the frequency of the knockdown resistance (kdr) L1014F and the ace-1 mutations, 2 markers associated respectively with pyrethroids and carbamate/organophosphate insecticide resistance.Results: A higher blood feeding rate was observed in An. gambiae compared to An. coluzzii as well as, a non-pro-nounced outdoor biting behavior in both species. The latter showed similar anthropophagic and sporozoite rates. However the analysis indicates a seasonal difference in the contribution of each species to malaria transmission associated with shifts in resting behaviour. Anopheles coluzzii females accounted for most of the detected infections: 86% in Alibori and 79% in Donga, during the dry season versus 14.4% and 21.2%, respectively for An. gambiae during the same period. This relationship was reversed in Donga during the rainy season (66% for An. gambiae against 34% for An. coluzzii). Results also indicated lower frequencies of kdr L1014F and ace-1 in An. coluzzii versus An. gambiae.Conclusion: Despite similarity in some parameters related to malaria transmission in both surveyed species, An. coluzzii is potentially a more important malaria vector because of high density in the region. It is also charac-terized by lower frequencies of the ace-1 mutation than is An. gambiae. The ongoing use of pirimiphos methyl (organophosphate) for IRS should continue to show a good impact in Alibori and Donga because of the very low level of the ace-1 mutation in both species

Quantification of the efficiency of treatment of Anopheles gambiae breeding sites with petroleum products by local communities in areas of insecticide resistance in the Republic of Benin

<p>Abstract</p> <p>Background</p> <p>The emergence of <it>Anopheles </it>populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for developing new insecticidal molecules or, improving the efficacy of existing insecticides or abandoning those to which resistance has emerged. The use of petroleum products (PP) against mosquito larvae had an immense success during early programmes of malaria control, but these compounds were abandoned and replaced in the 1950s by synthetic insecticides probably because of the high performances given by these new products. In the current context of vector resistance, it is important to elucidate the empirical use of PP by quantifying their efficiencies on resistant strains of <it>Anopheles</it>.</p> <p>Methods</p> <p>Larvae of <it>Anopheles </it>Ladji a local resistant strain were exposed to increasing concentrations of various PP (kerosene, petrol and engine oils) for 24 hours and the lethal activities recorded. The highest concentration (HiC) having no lethal activity (also referred as the NOEL or no effect level) and the lowest concentration (LoC₁₀₀) yielding 100% mortality were rated for each PP on the Ladji strain. Prior to laboratory analysis, KAP studies were conducted in three traditional communities were insecticide resistance is clearly established to confirm the use of PP against mosquitoes.</p> <p>Results</p> <p>Laboratory analysis of petrol, kerosene and engine oils, clearly established their lethal activities on resistant strains of <it>Anopheles </it>larvae. Contrary to existing references, this research revealed that exposed larvae of <it>Anopheles </it>were mostly killed by direct contact toxicity and not by suffocation as indicated in some earlier reports.</p> <p>Conclusion</p> <p>This research could serve as scientific basis to backup the empirical utilisation of PP on mosquito larvae and to envisage possibilities of using PP in some traditional settings where <it>Anopheles </it>have developed resistance to currently used insecticides.</p

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

<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

Insecticide resistance status in Anopheles gambiae in southern Benin

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

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

<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^R</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^R</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

Does the spillage of petroleum products in Anopheles breeding sites have an impact on the pyrethroid resistance?

<p>Abstract</p> <p>Background</p> <p>The emergence of <it>Anopheles </it>populations capable of withstanding lethal doses of insecticides has weakened the efficacy of most insecticide based strategies of vector control and, has highlighted the need for further studies on the mechanisms of insecticide resistance and the various factors selecting resistant populations of mosquitoes. This research targeted the analysis of breeding sites and the oviposition behaviour of susceptible and resistant populations of <it>Anopheles </it>in localities of spilled petroleum products. The aim was to establish the possible contribution of oil spillage in the selection of pyrethroid resistance in malaria vectors.</p> <p>Methods</p> <p><it>Anopheles </it>breeding sites were identified and the insecticide susceptibility of the <it>Anopheles gambiae </it>populations mapped in 15 localities of South Western Nigeria. The presence of oil particles as well as the turbidity, the dissolved oxygen and the pH of each identified breeding site was recorded. Data were cross-analysed to correlate the habitat types and the insecticide susceptibility status of emerging mosquitoes. The second phase of this study was basically a laboratory model to provide more information on the implication of the spillage of petroleum on the selection of pyrethroid resistance in <it>An. gambiae</it>.</p> <p>Results</p> <p>Moderate levels of resistance following exposure to permethrin-impregnated papers were recorded with the majority of <it>An. gambiae </it>samples collected in the South Western Nigeria. Data from this study established a link between the constituency of the breeding sites and the resistance status of the emerging <it>Anopheles</it>.</p> <p>Conclusion</p> <p>This study has revealed the segregational occupation of breeding habitats by pyrethroid resistant and susceptible strains of <it>An. gambiae </it>in south-western Nigeria. Compiled results from field and laboratory research point out clear relationships between oil spillage and pyrethroid resistance in malaria vectors. The identification of this factor of resistance could serve as strong information in the management of insecticide resistance in some West African settings.</p