Spread Of Insecticide Resistance Research Articles

Insecticide Resistance Profiling of Anopheles coluzzii and Anopheles gambiae Populations in the Southern Senegal: Role of Target Sites and Metabolic Resistance Mechanisms.

The emergence and spread of insecticide resistance among the main malaria vectors is threatening the effectiveness of vector control interventions in Senegal. The main drivers of this resistance in the Anopheles gambiae complex (e.g., An. gambiae and Anopheles coluzzii) remains poorly characterized in Senegal. Here we characterized the main target site and metabolic resistances mechanisms among the An. gambiae and An. coluzzii populations from their sympatric and allopatric or predominance area in Senegal. Larvae and pupae of An. gambiae s.l. were collected, reared to adulthood, and then used for insecticides susceptibility and synergist assays using the WHO (World Health Organisation) test kits for adult mosquitoes. The TaqMan method was used for the molecular characterization of the main target site insecticide resistance mechanisms (Vgsc-1014F, Vgsc-1014S, N1575Y and G119S). A RT-qPCR (Reverse Transcriptase-quantitative Polymerase Chaine Reaction) was performed to estimate the level of genes expression belonging to the CYP450 (Cytochrome P450) family. Plasmodium infection rate was investigated using TaqMan method. High levels of resistance to pyrethroids and DDT and full susceptibility to organophosphates and carbamates where observed in all three sites, excepted a probable resistance to bendiocarb in Kedougou. The L1014F, L1014S, and N1575Y mutations were found in both species. Pre-exposure to the PBO (Piperonyl butoxide) synergist induced a partial recovery of susceptibility to permethrin and full recovery to deltamethrin. Subsequent analysis of the level of genes expression, revealed that the CYP6Z1 and CYP6Z2 genes were over-expressed in wild-resistant mosquitoes compared to the reference susceptible strain (Kisumu), suggesting that both the metabolic resistance and target site mutation involving kdr mutations are likely implicated in this pyrethroid resistance. The presence of both target-site and metabolic resistance mechanisms in highly pyrethroid-resistant populations of An. gambiae s.l. from Senegal threatens the effectiveness and the sustainability of the pyrethroid-based tools and interventions currently deployed in the country. The Kdr-west mutation is widely widespread in An. coluzzii sympatric population. PBO or Duo nets and IRS (Indoor Residual Spraying) with organophosphates could be used as an alternative measure to sustain malaria control in the study area.

Ecological drivers of genetic connectivity for African malaria vectors Anopheles gambiae and An. arabiensis

Anopheles gambiae and An. arabiensis are major malaria vectors in sub-Saharan Africa. Knowledge of how geographical factors drive the dispersal and gene flow of malaria vectors can help in combatting insecticide resistance spread and planning new vector control interventions. Here, we used a landscape genetics approach to investigate population relatedness and genetic connectivity of An. gambiae and An. arabiensis across Kenya and determined the changes in mosquito population genetic diversity after 20 years of intensive malaria control efforts. We found a significant reduction in genetic diversity in An. gambiae, but not in An. arabiensis as compared to prior to the 20-year period in western Kenya. Significant population structure among populations was found for both species. The most important ecological driver for dispersal and gene flow of An. gambiae and An. arabiensis was tree cover and cropland, respectively. These findings highlight that human induced environmental modifications may enhance genetic connectivity of malaria vectors.

Vector genetics, insecticide resistance and gene drives: An agent-based modeling approach to evaluate malaria transmission and elimination.

Vector control has been a key component in the fight against malaria for decades, and chemical insecticides are critical to the success of vector control programs worldwide. However, increasing resistance to insecticides threatens to undermine these efforts. Understanding the evolution and propagation of resistance is thus imperative to mitigating loss of intervention effectiveness. Additionally, accelerated research and development of new tools that can be deployed alongside existing vector control strategies is key to eradicating malaria in the near future. Methods such as gene drives that aim to genetically modify large mosquito populations in the wild to either render them refractory to malaria or impair their reproduction may prove invaluable tools. Mathematical models of gene flow in populations, which is the transfer of genetic information from one population to another through migration, can offer invaluable insight into the behavior and potential impact of gene drives as well as the spread of insecticide resistance in the wild. Here, we present the first multi-locus, agent-based model of vector genetics that accounts for mutations and a many-to-many mapping cardinality of genotypes to phenotypes to investigate gene flow, and the propagation of gene drives in Anopheline populations. This model is embedded within a large scale individual-based model of malaria transmission representative of a high burden, high transmission setting characteristic of the Sahel. Results are presented for the selection of insecticide-resistant vectors and the spread of resistance through repeated deployment of insecticide treated nets (ITNs), in addition to scenarios where gene drives act in concert with existing vector control tools such as ITNs. The roles of seasonality, spatial distribution of vector habitat and feed sites, and existing vector control in propagating alleles that confer phenotypic traits via gene drives that result in reduced transmission are explored. The ability to model a spectrum of vector species with different genotypes and phenotypes in the context of malaria transmission allows us to test deployment strategies for existing interventions that reduce the deleterious effects of resistance and allows exploration of the impact of new tools being proposed or developed.

Resistance to insecticides in Anopheles gambiae complex in West Africa: A review of the current situation and the perspectives for malaria control

In West Africa, the Anopheles gambiae sensu lato (s.l.) complex mosquitoes are the main malaria vectors. Malaria control relies mainly on long-lasting insecticide-treated nets (LLINs) and to a lesser extent on indoor residual spray (IRS) of insecticides. Important progress has been made in terms of malaria morbidity and mortality in these past decades. Resistance to insecticides in An. gambiae s.l. was first attributed to their use in agriculture. Further, their use in public health increased selection pressure. Resistance has been recorded to the four main classes of insecticides used in public health in West Africa. However, the spread of insecticide resistance in West Africa is persistent due to the lack of management of resistance monitoring. Resistance to insecticides involves different mechanisms. Resistance to pyrethroids and organochlorines was caused by target modification such as some sodium channel mutations in addition to metabolic changes. Reduction of sensitivity to organophosphates and carbamates was caused by acetylcholinesterase mutations. Therefore, efforts should be made as quickly as possible to develop novel vector control techniques. Furthermore, it is also suggested to combine environmental, chemical, and biological management. In this review, we outline the insecticide resistance of Anopheles in West Africa and the current alternative tools for effective mosquito vector control.

Indoor residual spraying for malaria control in sub-Saharan Africa 1997 to 2017: an adjusted retrospective analysis

BackgroundIndoor residual spraying (IRS) is a key tool for controlling and eliminating malaria by targeting vectors. To support the development of effective intervention strategies it is important to understand the impact of vector control tools on malaria incidence and on the spread of insecticide resistance. In 2006, the World Health Organization (WHO) stated that countries should report on coverage and impact of IRS, yet IRS coverage data are still sparse and unspecific. Here, the subnational coverage of IRS across sub-Saharan Africa for the four main insecticide classes from 1997 to 2017 were estimated.MethodsData on IRS deployment were collated from a variety of sources, including the President’s Malaria Initiative spray reports and National Malaria Control Programme reports, for all 46 malaria-endemic countries in sub-Saharan Africa from 1997 to 2017. The data were mapped to the applicable administrative divisions and the proportion of households sprayed for each of the four main insecticide classes; carbamates, organochlorines, organophosphates and pyrethroids was calculated.ResultsThe number of countries implementing IRS increased considerably over time, although the focal nature of deployment means the number of people protected remains low. From 1997 to 2010, DDT and pyrethroids were commonly used, then partly replaced by carbamates from 2011 and by organophosphates from 2013. IRS deployment since the publication of resistance management guidelines has typically avoided overlap between pyrethroid IRS and ITN use. However, annual rotations of insecticide classes with differing modes of action are not routinely used.ConclusionThis study highlights the gaps between policy and practice, emphasizing the continuing potential of IRS to drive resistance. The data presented here can improve studies on the impact of IRS on malaria incidence and help to guide future malaria control efforts.

Use of alternative bioassays to explore the impact of pyrethroid resistance on LLIN efficacy

BackgroundThere is substantial concern that the spread of insecticide resistance will render long-lasting insecticide-treated nets (LLINs) ineffective. However, there is limited evidence supporting a clear association between insecticide resistance and malaria incidence or prevalence in the field. We suggest that one reason for this disconnect is that the standard WHO assays used in surveillance to classify mosquito populations as resistant are not designed to determine how resistance might impact LLIN efficacy. The standard assays expose young, unfed female mosquitoes to a diagnostic insecticide dose in a single, forced exposure, whereas in the field, mosquitoes vary in their age, blood-feeding status, and the frequency or intensity of LLIN exposure. These more realistic conditions could ultimately impact the capacity of “resistant” mosquitoes to transmit malaria.MethodsHere, we test this hypothesis using two different assays that allow female mosquitoes to contact a LLIN as they host-seek and blood-feed. We quantified mortality after both single and multiple exposures, using seven different strains of Anopheles ranging in pyrethroid resistance intensity.ResultsWe found that strains classified as 1×-resistant to the pyrethroid insecticide deltamethrin in the standard WHO assay exhibited > 90% mortality over 24 h following more realistic LLIN contact. Mosquitoes that were able to blood-feed had increased survival compared to their unfed counterparts, but none of the 1×-resistant strains survived for 12 days post-exposure (the typical period for malaria parasite development within the mosquito). Mosquitoes that were 5×- and 10×-resistant (i.e. moderate or high intensity resistance based on the WHO assays) survived a single LLIN exposure well. However, only about 2–3% of these mosquitoes survived multiple exposures over the course of 12 days and successfully blood-fed during the last exposure.ConclusionsThese results suggest that the standard assays provide limited insight into how resistance might impact LLIN efficacy. In our laboratory setting, there appears little functional consequence of 1×-resistance and even mosquitoes with moderate (5×) or high (10×) intensity resistance can suffer substantial reduction in transmission potential. Monitoring efforts should focus on better characterizing intensity of resistance to inform resistance management strategies and prioritize deployment of next generation vector control products.

Effects of insecticide resistance and exposure on Plasmodium development in Anopheles mosquitoes.

The spread of insecticide resistance in anopheline mosquitoes is a serious threat to the success of malaria control and prospects of elimination, but the potential impact(s) of insecticide resistance or sublethal insecticide exposure on Plasmodium-Anopheles interactions are poorly understood. Only a few studies have attempted to investigate such interactions, despite their clear epidemiological significance for malaria transmission. This short review provides an update on our understanding of the interactions between insecticide resistance and exposure and Plasmodium development, focusing on the mechanisms which might underpin any interactions, and identifying some key knowledge gaps.

Exploring the impact of glutathione S-transferase (GST)-based metabolic resistance to insecticide on vector competence of Anopheles funestus for Plasmodium falciparum.

Background: Malaria control heavily relies on insecticide-based interventions against mosquito vectors. However, the increasing spread of insecticide resistance is a major threat. The extent to which such resistance, notably metabolic resistance, interferes with the development of the Plasmodium parasite and its impact on overall malaria transmission remains poorly characterized. Here, we investigated whether glutathione S-transferase-based resistance could influence Plasmodium falciparum development in Anopheles funestus. Methods: Anopheles funestus females were infected with P. falciparum gametocytes and midguts were dissected at day 7 post infection for detection/quantification of oocysts. Infection parameters were compared between individual with different L119F-GSTe2 genotypes, and the polymorphism of the GSTe2 gene was analyzed in infected and uninfected mosquito groups. Results: Overall, 403 mosquitoes were dissected and genotyped. The frequency of the L119F-GSTe2 resistance allele was significantly higher in non-infected (55.88%) compared to infected (40.99%) mosquitoes (Fisher's exact test, P<0.0001). Prevalence of infection was significantly higher in heterozygous and homozygous susceptible genotypes (P<0.001). However, homozygous resistant and heterozygous mosquitoes exhibited significantly higher infection intensity (P<0.01). No association was observed between the GSTe2 polymorphism and the infection status of mosquitoes. Conclusion: Altogether, these results suggest that GSTe2-based metabolic resistance may affect the vectorial competence of resistant An. funestus mosquitoes to P. falciparum infection, by increasing its permissiveness to Plasmodium infection.

Exploring the impact of glutathione S-transferase (GST)-based metabolic resistance to insecticide on vector competence of Anopheles funestus for Plasmodium falciparum

Background: Malaria control heavily relies on insecticide-based interventions against mosquito vectors. However, the increasing spread of insecticide resistance is a major threat. The extent to which such resistance, notably metabolic resistance, influences the development of the Plasmodium parasite and its impact on overall malaria transmission remains poorly characterized. Here, we investigated whether glutathione S-transferase-based resistance could influence Plasmodium falciparum development in Anopheles funestus. Methods: Anopheles funestus females were infected with P. falciparum gametocytes and midguts were dissected at day 7 post infection for detection/quantification of oocysts. Infection parameters were compared between individuals with different L119F-GSTe2 genotypes, and the polymorphism of the GSTe2 gene was analyzed in infected and uninfected mosquito groups. Results: Overall, 403 An. funestus mosquitoes were dissected and genotyped. The frequency of the L119F-GSTe2 resistance allele was significantly higher in non-infected (55.88%) compared to infected (40.99%) mosquitoes (Fisher's exact test, P<0.0001). Prevalence of infection was significantly higher in heterozygous and homozygous susceptible genotypes (P<0.001). However, homozygous resistant and heterozygous mosquitoes exhibited significantly higher infection intensity (P<0.01). No association was observed between the GSTe2 polymorphism and the infection status of mosquitoes. Conclusion: Altogether, these results suggest that GSTe2-based metabolic resistance may affect the vectorial competence of resistant An. funestus mosquitoes to P. falciparum infection, by possibly increasing its permissiveness to Plasmodium infection.

Diagnostic dose determination and efficacy of chlorfenapyr and clothianidin insecticides against Anopheles malaria vector populations of western Kenya

BackgroundMalaria vector control is dependent on chemical insecticides applied to walls by indoor residual spraying or on long-lasting insecticidal nets. The emergence and spread of insecticide resistance in major malaria vectors may compromise malaria control and elimination efforts. The aim of this study was to estimate a diagnostic dose for chlorfenapyr (class: pyrrole) and clothianidin (class: neonicotinoid) and assess the baseline susceptibility of three major Anopheles malaria vectors of western Kenya to these two insecticides.MethodsThe Centers for Disease Control and Prevention (CDC) bottle assay was used to determine the diagnostic doses of chlorfenapyr and clothianidin insecticides against the susceptible Kisumu strain of Anopheles gambiae. Probit analysis was used to determine the lethal doses at which 50% (LD50) and 99% (LD99) of the susceptible mosquitoes would be killed 24, 48 and 72 h following exposure for 1 h. Insecticidal efficacy of chlorfenapyr, clothianidin and the pyrethroid deltamethrin was then evaluated against field collected female Anopheles mosquitoes sampled from Nyando, Bumula and Ndhiwa sub-Counties in western Kenya. Members of Anopheles funestus and An. gambiae complexes were identified using polymerase chain reaction (PCR).ResultsThe determined diagnostic doses of chlorfenapyr and clothianidin insecticides were 50 µg/bottle and 150 µg/bottle, respectively, for An. gambiae, Kisumu strain. When exposed to the diagnostic dose of each insecticide, Anopheles malaria vector populations in western Kenya were susceptible to both insecticides with 100% mortality observed after 72 h. Mortality of mosquitoes exposed to deltamethrin increased over time but did not reach 100%. Mortality of Anopheles arabiensis from Nyando exposed to deltamethrin was 83% at 24 h, 88% at 48 h and 94.5% at 72 h while An. funestus from Ndhiwa was 89% at 24 h, 91.5% at 48 h and 94.5% at 72 h.ConclusionMosquitoes of western Kenya, despite being resistant to pyrethroids, are susceptible to chlorfenapyr and clothianidin. Field evaluations of the formulated product are needed.

Bio-efficacy and physical integrity of piperonylbutoxide coated combination net (PermaNet\xae 3.0) against pyrethroid resistant population of Anopheles gambiae s.l. and Culex quinquefasciatus mosquitoes in Ethiopia

BackgroundPermaNet® 3.0 is a deltamethrin-treated combination long-lasting insecticidal net with the addition of synergist piperonylbutoxide (PBO) on its roof section. It is designed to overcome the challenge posed by pyrethroid resistant vector populations against mainstream long-lasting insecticidal nets impregnated with pyrethroids only. The objective of this study was to determine insecticide resistance status of Anopheline and Culicine mosquitoes, to evaluate the bio-efficacy of PermaNet® 3.0 nets and to assess household factors affecting the physical integrity of PermaNet® 3.0 after 3 years of use.MethodsInsecticide susceptibility test was conducted using the WHO tube test. Bio-activity of PermaNet® 3.0 samples was evaluated using the WHO cone bioassay. Cross-sectional survey was conducted on 150 randomly selected households from two districts to determine household factors affecting net utilization. One hundred fifty PermaNet® 3.0 nets were randomly collected from the community with replacement after 3 years of deployment and physical integrity of each net was assessed.ResultsBoth Anopheles gambiae sensu lato and Culex quinquefasciatus developed resistance against permethrin and deltamethrin. However, following pre-exposure to synergist PBO the susceptibility of mosquito population increased to both permethrin (from 39% without to 92% with PBO against An. gambiae and from 28% without to 94% with PBO against Culex quinquefasciatus) and deltamethrin (from 52% without to 99% with PBO against An. gambiae and from 43% without to 98% with PBO against Culex quinquefasciatus). Eighty percent (80%) mortality was recorded in wild population of An. gambiae s.l. exposed to unused PermaNet® 3.0, but its bioactivity subsequently declined as washing frequency increased from 0 to 20. The PBO coated roof section of unused PermaNet® 3.0 resulted in higher mosquito mortality (100%) compared to the side panels without PBO (85%). House structure, cooking and washing habits, and damage due to household pests were cited as determinants associated with bed net deterioration. Bed net proportionate hole index (pHI) was ranged from 0 to 6064. Of the 150 PermaNet® 3.0 nets assessed 80, 29 and 41 were considered as ‘good’, ‘acceptable’ and ‘too torn’, respectively.ConclusionsThe bio-efficacy evaluation of PermaNet® 3.0 from Jimma area, southwestern Ethiopia showed moderate efficacy against pyrethroid resistant population of An. gambiae and Culex quinquefasciatus. Thus, NMCPs in parallel to deployment of LLINs, should implement timely insecticide resistance management and integrated vector management strategies to slowdown the evolution and further spread of insecticide resistance. Household factors such as, housing conditions, open flame fire used for cooking and rodent attack were identified as factors contributing to the observed reduced bed net physical integrity in the study area. Universal coverage of bed nets should be accompanied with community awareness creation and training on net utilization and handling.

Low Knockdown Resistance Frequency In Anopheles Gambiae Complex Resistant To Some Who-Approved Insecticides In Selected Areas Of Lagos State, South-West, Nigeria

Background: Decline in malaria prevalence is usually attributed to the efficient vector control strategies implemented in such endemic areas. The spread of insecticide resistance in Anopheles mosquitoes is a major drawback to the gains in malaria vector control. Here we assessed the susceptibility status of Anopheles gambiae s.l. to some selected WHO-approved insecticides and frequency of knockdown resistance gene in two Local Government Areas of Lagos State, Nigeria. Methods: Three to five days old adult female mosquitoes were exposed to deltamethrin, permethrin and bendocarb. Knockdown time was recorded every ten minutes and % mortality taken at 24hr post exposure. KDT50 and KDT95 were determined using probit regression analysis. The resistant mosquitoes were used for species identification by Polymerase Chain Reaction (PCR) assays and characterized for the knockdown resistance (kdr) mutation by allele-specific PCR (AS-PCR). Results: The results of twenty-four hour post exposure mortality rate showed that mosquitoes sampled at differ-ent locations were resistant to permethrin (39% mortality for Lagos Mainland L.G.A.; 45% for Kosofe L.G.A.), del-tamethrin (51% for Lagos Mainland L.G.A; 68% for Kosofe L.G.A), but susceptible to bendiocarb (100% for Lagos Mainland L.G.A.; 99% for Kosofe L.G.A.). The KDT50 varied from 34 minutes in bendiocarb for both LGAs to 190 minutes in deltamethrin for Kosofe and 119 minutes in permethrin for Lagos Mainland. In all insecticides tested for both LGAs, KDT95 was greater than 60 minutes. Only the 1014F kdr mutation was detected. The overall kdr frequen-cy was low (1.79%). There was no significant association between the presence of the 1014F kdr allele and ability of Anopheles mosquitoes to survive exposure to the tested insecticides (P &gt; 0.05). Conclusion: This level of resistance to WHO approved insecticides is a threat to control programs and can have significant operational impact, if action is not taken.

Progress and challenges of global malaria elimination

Recently, the global malaria control has achieved remarkable results, and the epidemic map of malaria has gradually shrinked. However, in the past two years, the number of malaria deaths remained at a high level, and the incidence of malaria has even risen, leading to the stagnant of malaria elimination. The main reasons include lacking of the well monitoring and response system, sensitivity declining of antimalarial drugs, the spread of insecticide resistance, and the reduction of financial support. This paper introduces the progress and challenges of global malaria elimination, summarizes the current strategies and major interventions, and provides the corresponding response.

Exploring the impact of glutathione S-transferase (GST)-based metabolic resistance to insecticide on vector competence of Anopheles funestus for Plasmodium falciparum

Background:Malaria control heavily relies on insecticide-based interventions against mosquito vectors. However, the increasing spread of insecticide resistance is a major threat. The extent to which such resistance, notably metabolic resistance, interferes with the development of thePlasmodiumparasite and its impact on overall malaria transmission remains poorly characterized. Here, we investigated whether glutathione S-transferase-based resistance could influencePlasmodium falciparumdevelopment inAnopheles funestus.Methods:Anopheles funestusfemales were infected withP. falciparumgametocytes and midguts were dissected at day 7 post infection for detection/quantification of oocysts. Infection parameters were compared between individual with different L119F-GSTe2 genotypes, and the polymorphism of the GSTe2 gene was analyzed in infected and uninfected mosquito groups.Results:Overall, 403 mosquitoes were dissected and genotyped. The frequency of the L119F-GSTe2 resistance allele was significantly higher in non-infected (55.88%) compared to infected (40.99%) mosquitoes (Fisher's exact test, P&lt;0.0001). Prevalence of infection was significantly higher in heterozygous and homozygous susceptible genotypes (P&lt;0.001). However, homozygous resistant and heterozygous mosquitoes exhibited significantly higher infection intensity (P&lt;0.01). No association was observed between the GSTe2 polymorphism and the infection status of mosquitoes.Conclusion:Altogether, these results suggest that GSTe2-based metabolic resistance may affect the vectorial competence of resistantAn. funestusmosquitoes toP. falciparuminfection, by increasing its permissiveness toPlasmodiuminfection.

Rapid spread of double East- and West-African kdr mutations in wild Anopheles coluzzi from Côte d’Ivoire

Malaria morbidity and mortality rates in Sub-Saharan Africa are increasing. The scale-up of long-lasting insecticidal nets and indoor residual spraying have been the major contributors to the decrease of malaria burden. These tools are now threatened by insecticide resistance in malaria vectors, which is spreading dramatically. After two different real-time polymerase chain reaction molecular characterizations carried out on 70 mosquitoes sampled in the locality of Elibou in southern Côte d’Ivoire, results revealed that 9 mosquitoes from Anopheles coluzzi harbored the double East- and West-African knockdown resistance mutations. In the previous year, only 1 mosquito out of 150 sampled from 10 regions of the country had the same genotype. These results show the rapid spread of insecticide resistance in malaria vectors and highlight the urgent need to diversify the methods of vector control in order to avoid the failure of insecticide-based vector control tools which may favor malaria fatalities.

A cluster of CYP6 gene family associated with the major quantitative trait locus is responsible for the pyrethroid resistance in Culex pipiens pallen.

The emergence and rapid spread of insecticide resistance in several mosquito species has become a significant obstacle in management of mosquito-borne diseases, including deltamethrin resistance in Culex pipiens pallens. Previous study identified a major deltamethrin resistance quantitative trait locus (DR-6) that alone explained 62% of the genetic variance. In this study, the marker L4B1.102 and L4B1.175 associated with the DR-6 were characterized. We searched for potential candidate genes in the flank region of two markers in the genome sequence and showed that a cluster of CYP6 cytochrome P450 genes (CYP6BB4, CYP6BB3, CYP6CC2, CYP6P14, CYP6BZ2, CYP6AA9, CYP6AA8, CYP6AA7) was in the vicinity of DR-6. Significant differences in the expression of these P450s in the larval and adult stages were identified in the resistant strains compared with the susceptible strain. For CYP6AA9 and CYP6BB4, the correlation analysis showed a highly positive correlation between relative gene expression quantification and the resistance level in different strains. Knockdown of CYP6BB4 increased the sensitivity of mosquitoes to deltamethrin. We identified that the deltamethrin resistance was in a cluster of CYP6 genes in C.pipiens pallens, and CYP6BB4 may play a significant role in the development of deltamethrin resistance.

Evaluation of toxicity of clothianidin (neonicotinoid) and chlorfenapyr (pyrrole) insecticides and cross-resistance to other public health insecticides in Anopheles arabiensis from Ethiopia

BackgroundInsecticide-based interventions play an integral role in malaria vector control. However, the continued spread of insecticide resistance threatens to undermine progress made thus far and may ultimately lead to operational failure of current control measures. Clothianidin and chlorfenapyr both have unique modes of action and have expanded the number of insecticide classes available to vector control programmes. Prior to field use, it is imperative to establish their toxicity against local mosquito populations and evaluate potential cross-resistance with other chemicals used contemporarily or historically. The aim of this study was to determine the diagnostic doses of clothianidin and chlorfenapyr and their efficacies against Anopheles arabiensis, the predominant Ethiopian malaria vector species.MethodsA range of doses of clothianidin and chlorfenapyr were tested, using modified WHO susceptibility tests and CDC bottle bioassays, respectively, against an Ethiopian susceptible laboratory strain and a wild population of An. arabiensis collected from Oromia Region, Ethiopia. Cross-resistance to other public health insecticides: carbamates (bendiocarb and propoxur), organophosphate (malathion) and pyrethroids (deltamethrin and permethrin), was assessed in the same mosquito populations using CDC bottle bioassays.ResultsComplete mosquito mortality was observed with the laboratory strain using the recommended diagnostic doses for clothianidin (2%/filter paper) and chlorfenapyr (100 µg/bottle). The field population was resistant to malathion (83% mortality), capable of surviving 2×, 5× and 10× the diagnostic dose of both deltamethrin and permethrin, but susceptible to bendiocarb and propoxur. The field population of An. arabiensis was significantly more susceptible to clothianidin, reaching 100% mortality by day 2 compared to the laboratory strain (100% mortality by day 3). In contrast, the wild population was less susceptible to chlorfenapyr, with the highest mortality of 99% at 72 h using 200 µg/bottle compared to the laboratory colony, which reached complete mortality at 50 µg/bottle by 24 h.ConclusionsThe putative diagnostic doses of clothianidin and chlorfenapyr are appropriate for monitoring resistance in An. arabiensis from Ethiopia. The unique modes of action and an absence of cross-resistance render clothianidin and chlorfenapyr potential candidates for inclusion in the National Malaria Control Programme vector control efforts, particularly in areas with high pre-existing or emergent resistance to other insecticide classes.

Rapid spread of double East- and West-African kdr mutations in wild Anopheles coluzzi from Côte d'Ivoire.

Malaria morbidity and mortality rates in Sub-Saharan Africa are increasing. The scale-up of long-lasting insecticidal nets and indoor residual spraying have been the major contributors to the decrease of malaria burden. These tools are now threatened by insecticide resistance in malaria vectors, which is spreading dramatically. After two different real-time polymerase chain reaction molecular characterizations carried out on 70 mosquitoes sampled in the locality of Elibou in southern Côte d'Ivoire, results revealed that 9 mosquitoes from Anopheles coluzzi harbored the double East- and West-African knockdown resistance mutations. In the previous year, only 1 mosquito out of 150 sampled from 10 regions of the country had the same genotype. These results show the rapid spread of insecticide resistance in malaria vectors and highlight the urgent need to diversify the methods of vector control in order to avoid the failure of insecticide-based vector control tools which may favor malaria fatalities.

Insecticide Resistance Status and Mechanisms of Anopheles sinensis (Diptera: Culicidae) in Wenzhou, an Important Coastal Port City in China.

Although scaled-up interventions and effective control efforts have drastically reduced malaria morbidity and mortality, malaria remains a serious threat to public health worldwide. Anopheles sinensis Wiedemann 1828 is a historically important vector of Plasmodium vivax (Haemosporida: Plasmodiidae) malaria in China. Insecticide resistance has become a major obstacle to vector-borne disease control. However, little is known about the insecticide resistance of An. sinensis in Wenzhou, an important coastal port city in Zhejiang province, China. The aim of this study was to examine insecticide resistance and mechanisms in An. sinensis field mosquito populations. Evidence of multiple insecticide resistance was found in An. sinensis adult female populations. Medium to high frequencies of target site kdr together with fixed ace-1 mutations was detected in both the Ruian and Yongjia populations. Both populations showed an association between kdr L1014 mutation and resistance phenotype when tested against deltamethrin and DDT. Significantly different metabolic enzyme activities were found between the susceptible laboratory strain and field-collected mosquitoes from both Ruian and Yongjia. Both field collected An. sinensis populations exhibited significantly higher P450 enzyme activity compared with the laboratory strain, while the field-collected resistant mosquitoes exhibited various GST and COE enzyme activities. These results indicate multiple resistance mechanisms in An. sinensis field populations. Effective implementation of insecticide resistance management strategies is urgently needed. The data collected in this study will be valuable for modeling insecticide resistance spread and vector-control interventions.

Indoor Residual Spraying for Malaria Control in Sub-Saharan Africa 1997 to 2017: An Adjusted Retrospective Analysis

Background: Indoor residual spraying (IRS) is a key tool for controlling and eliminating malaria by targeting vectors. To support the development of effective intervention strategies it is important to understand the impact of vector control tools on malaria incidence and on the spread of insecticide resistance. In 2006, the World Health Organization (WHO) stated that countries should report on coverage and impact of IRS, yet IRS coverage data are still sparse and unspecific. Methods: Data on IRS deployment were collated from a variety of sources, including the President's Malaria Initiative spray reports and National Malaria Control program reports, for all 46 malaria-endemic countries in Sub-Saharan Africa from 1997 to 2017. The data were mapped to the applicable administrative divisions and the proportion of households sprayed for each of the four main insecticide classes; carbamates, organochlorines, organophosphates and pyrethroids was calculated. Findings: The number of countries implementing IRS has increased considerably although coverage in-country is often low, particularly in West Africa. From 1997 to 2010 there was an expansion of IRS using organochlorines (DDT) and pyrethroids. From 2011, carbamates partly replaced these insecticides and organophosphate use increased from 2013 onwards, however, policy recommendations to rotate between insecticide classes were rarely followed. Interpretation: Our results highlight trends in IRS coverage and insecticide choice in sub Saharan Africa. The data presented here can improve studies on the impact of IRS on malaria incidence and on insecticide resistance development and help to guide future malaria control efforts. Funding Statement: Wellcome Trust grant 108440/Z/15/Z. Declaration of Interests: The authors declare no conflict of interest. Ethics Approval Statement: Not Required.