malERA: An updated research agenda for insecticide and drug resistance in malaria elimination and eradication
Janet Hemingway and colleagues examine progress in research on insecticide and drug resistance in malaria elimination and eradication and propose a research agenda.
Published in the journal:
. PLoS Med 14(11): e32767. doi:10.1371/journal.pmed.1002450
Category:
Collection Review
doi:
https://doi.org/10.1371/journal.pmed.1002450
Summary
Janet Hemingway and colleagues examine progress in research on insecticide and drug resistance in malaria elimination and eradication and propose a research agenda.
Summary points
Since 2011, significant progress has been made in understanding resistance. Surveillance has been expanded and improved in many malaria-endemic countries and there is a better understanding of the genetic basis of resistance, identifying some molecular markers that can be used to track its emergence and spread. Better tools to measure and manage the intensity of resistance are available.
However, our response to increases in the prevalence and intensity of resistance has been slow and reactive. A promising pipeline of new vector control tools and therapeutics is in development, but all actors in the malaria community need to plan proactively how to implement, integrate, and evaluate these products.
Quantifying the public health impact of resistance has been difficult, particularly for insecticides. For both insecticides and drugs, defining the minimum essential evidence required for policy makers to manage resistance and ensuring that programs employ rigorous quality assurance in collecting and managing these data are critical.
As malaria control increases, the selection pressure on the parasite or mosquito vector increases. Strategies for resistance management are therefore crucial for all stages of elimination. Countries need to allocate funding and human resources to effectively manage the threat of resistance and sustain the gains achieved to date.
This paper reviews the current knowledge base and identifies research priorities addressing resistance to drugs and insecticides. It is a result of a unique collaborative effort of experts in drug and insecticide resistance brought together for the malERA Refresh process.
Introduction and rationale
Over the past decade, unprecedented progress has been made in reducing malaria morbidity and mortality [1]. However, growing resistance to the first-line treatment for P. falciparum malaria, artemisinin-based combination therapies (ACTs), and the insecticides used to suppress mosquito vectors threaten the sustainability of recent gains in malaria control and longer-term prospects for elimination.
Vector control and antimalarial treatment depend on a limited armamentarium, and when single drugs and insecticides are widely deployed, selection pressure is intense and the emergence of resistant parasites and mosquitoes is inevitable.
Drug and insecticide resistance were crosscutting issues in the original malERA (Malaria Eradication Research Agenda) series in 2011 [2]. However, the parasite and vector communities rarely interact. The increasing urgency of these issues and the contrasting operational responses warranted a dedicated panel in the malERA Refresh process. The failure of drug treatment has human consequences: recurrent parasitaemia, severe malaria, anaemia, and associated morbidity and mortality. In the early 2000s, resistance to single antimalarials led to policy changes recommending deployment of ACTs [3]. In contrast, resistance to the most widely used class of insecticide, pyrethroids, was first documented in the 1980s, but pyrethroid monotherapies still dominate current control efforts [4].
This paper aims to review developments in drug and insecticide resistance over the past 5 years (Box 1), discuss gaps in knowledge, and identify key research priorities (Box 2).
Box 1. Progress over the past 5 years in drug and insecticide resistance research
A promising pipeline of new therapeutics, insecticides, and noninsecticidal vector control tools is in development, largely due to the work of the Medicines for Malaria Venture (MMV) and the Innovative Vector Control Consortium (IVCC)
Recognition of the impact and importance of drug and insecticide resistance with the creation of the WHO Global Plan for Insecticide Resistance Management in malaria vectors (GPIRM) and WHO Global Plan for Artemisinin Resistance Containment (GPARC)
Identification of genes and molecular markers associated with drug and insecticide resistance
Improved understanding of resistance mechanisms in parasite and vector populations
Global databases to monitor drug and insecticide resistance
Development of new tools to study resistance in vivo and in vitro, e.g., ring-stage survival assay, parasite clearance estimator, human blood-stage challenge studies for drug resistance, and bioassays that measure the intensity of insecticide resistance
Box 2. Research and development agenda for drug and insecticide resistance
Crosscutting issues for drug and insecticide resistance
Applied research.
Use in vitro, in vivo, and mathematical models to identify new combinations of drugs and insecticides, and understand how mechanisms of action and mechanisms of resistance inform this
Determine which conditions are optimal for the emergence and spread of drug and insecticide resistance and how these can be minimised
Evaluate whether resistance management strategies can restore susceptibility to drugs and insecticides
Evaluate how new intervention types/paradigms should be introduced and assessed to limit the selection of resistant phenotypes
Evaluate the optimal surveillance systems for resistance and determine the appropriate data that must be collected (including technical approach, frequency, geography, and temporal–spatial factors)
Determine and validate the relationships between molecular markers and parasite/vector resistance phenotypes in different transmission settings
Policy and advocacy.
Develop a framework to cost-elimination strategies that accounts for resistance management practices and increasing cost per case of malaria/malaria death averted and identify sources of funding for these strategies
Agree on the process and minimum data required by the normative bodies to enable a new drug or insecticide product to complete the route to market
Devise market strategies and incentives to ensure a mix of drug and insecticide products remains available and is used strategically to manage resistance
Assess which decision-support systems can efficiently and rationally be adapted to drug and insecticide policies
Determine the minimum dataset required to guide drug and insecticide resistance management and the level of evidence required to switch to new drug or insecticide strategies
Insecticide resistance
Analyse the most cost-effective ways of slowing the spread and emergence of insecticide resistance (e.g., by using a combination of interventions, spatial mosaics, or mixtures of insecticides)
Determine which spatial and temporal scale insecticide resistance management strategies should be carried out
Study how much insecticide resistance has a negative impact on mosquito fitness survival or parasite development in the mosquito and investigate how this compares for different active ingredients
Develop a method to assess the age of resistant mosquitoes
Define the optimal use of bioassays and molecular markers to accurately predict the efficacy of vector control in relation to insecticide resistance
Study the mechanisms of mosquito behavioural resistance and assess if this is sustained across generations
Assess which novel, noninsecticidal tools for controlling mosquito populations would help to slow or prevent the emergence of resistance or restore susceptibility
Drug resistance
Evaluate if the timing of community-based prevention, e.g., mass drug administration, can be optimised to reduce the risk of emerging drug resistance
Investigate why drugs such as quinine are less likely to develop resistance and use this knowledge for future drug development
Determine which approaches are most sensitive and specific to determine true drug treatment efficacy (e.g., molecular correction) in P. falciparum and P. vivax parasites
Define what studies are required by policy makers to evaluate the use of multiple therapies
Define the minimal criteria for inclusion of existing and new drugs in multiple agent regimes (e.g., efficacy, resistance, pharmacokinetic factors, and drug–drug interactions) and whether these criteria change in different programmatic modes
Study the extent to which human immunity masks the presence of drug resistance, especially resistance to artemisinins
Methods
The findings presented in this paper result from an extensive literature review of published and unpublished materials and the deliberations of the 2015 malERA Refresh Consultative Panel on Insecticide and Drug Resistance. Electronic databases were systematically searched for published literature between January 1, 2010, and November 2, 2015, without language limitations. Panellists were invited to recommend additional literature. A 2-day workshop was held with the majority of the panel members, including specialists from basic science and product development, field researchers, and WHO representatives. The panel broke into 2 working groups to identify the problems that need to be solved in insecticide and drug resistance and what research is needed to address these problems. Each group fed back to the plenary session, in which further robust discussions and input occurred. This helped refine the opportunities and gap areas in which research is needed. The final findings were arrived at with input from all panellists and several iterations of the manuscript.
What do we know about resistance?
Insecticides for malaria vector control are limited to pyrethroids for long-lasting insecticidal nets (LLINs) and pyrethroids, organochlorines, organophosphates, and carbamates for indoor residual spraying (IRS). Vector resistance has been detected across Africa to all insecticide classes. However, resistance to the pyrethroids is the most widespread [5]. In Asia, insecticide resistance is common in some Anopheles species [6]. Sixty countries have reported resistance to at least 1 insecticide, but the scale of the problem is likely to be much greater [7].
Despite ubiquitous pyrethroid resistance in some areas, millions of pyrethroid-impregnated nets are distributed annually. Once distributed, these nets can contribute to the selection of resistant vectors for the duration of their 3-year life. In Burkina Faso, the intensity of the pyrethroid resistance seen in A. gambiae increased 10-fold in a single year [8], and this trend is apparent in multiple locations throughout Africa [9]. A. funestus also exhibits resistance to multiple insecticides at increasing intensities [10–12]. Proactive defensive strategies are critical to reducing the spread and emergence of resistant phenotypes and preventing broad-spectrum cross resistance to multiple insecticides.
In the case of the antimalarials chloroquine and sulfadoxine-pyrimethamine (SP), resistant P. falciparum and P. vivax parasites evolved in the Greater Mekong Subregion (GMS) and the island of Papua and South America, respectively [13,14]. Retrospective analysis of molecular markers showed resistant P. falciparum parasites spread from Southeast Asia foci across Asia and throughout Africa over several decades [15–18]. ACTs were promoted to prevent or retard the selection of resistance by simultaneously administering 2 drug components with different modes of action [19]. However, resistance to artemisinins and their partner drugs is spreading and emerging independently among P. falciparum populations in the GMS [20–23].
Identifying resistance
Two main mechanisms of insecticide resistance have been identified: target site mutations (such as kdr and ace) [24,25] and metabolic resistance involving mutation, duplication, or altered regulation of enzymes and transporters that increase insecticide metabolism or excretion. Metabolic resistance has greater implications for malaria vector control because the efficacy of a range of insecticides is usually affected [5,26].
Routine monitoring of insecticide susceptibility uses phenotypic bioassays that expose live mosquitoes to a single dose of a given insecticide over a fixed time period and measure mortality. The results are highly variable; hence, more laborious methods utilising a range of insecticide concentrations may be needed [27]. These assays have local utility but are often logistically challenging. Larger numbers of mosquitoes can be screened using molecular techniques, although it is unclear under what conditions validated molecular markers could serve as a replacement for phenotypic assays or if this might be appropriate for malaria control programmes [28].
The mechanisms of insecticide resistance can manifest as major changes in the insect nervous system or metabolome. Resistance may have an effect on insect longevity, mating competitiveness, and vectorial capacity [29,30]. Alongside physiological resistance, there is potentially also behavioural resistance, as increased mosquito numbers that bite or rest outdoors have been observed. There is limited evidence on the genetic basis of behavioural resistance, but determining whether vector control interventions are selecting a heritable trait warrants further research [31].
Resistance to artemisinins is assessed in clinical studies by measuring the parasite clearance in a patient in the first several days after treatment [32]. A lab-based assay that correlates with the in vivo parasite response to artemisinins has also been validated [33]. Mutations in the propeller domain of Kelch 13 (PF3D7_1343700) (K13) were identified as a major determinant of artemisinin resistance and may be reliable molecular markers in the GMS [34,35]. Outside the GMS, parasites with K13 mutant alleles are present in many areas at low levels; there is currently no molecular evidence to suggest that these alleles are being selected [22,36–38]. More than 100 K13 mutant alleles have been reported outside of Southeast Asia [22,38–40], but none have yet been associated with the slow-clearing phenotype [41]. One hypothesis is that artemisinin resistance may require additional genetic determinants in these locations to allow selection of K13 mutant parasites that exhibit the slow-clearing phenotype in vivo [20,42]. Nevertheless, the adoption of molecular markers to monitor drug resistance has been much faster than markers to assess insecticide resistance.
Molecular markers correlated with resistance to nonartemisinin antimalarials have also been identified. Polymorphisms or multicopy numbers in the P. falciparum chloroquine resistance transporter (pfcrt) and P. falciparum multidrug resistance 1 (pfmdr1) genes have been associated with resistance to chloroquine and mefloquine [43,44] and polymorphisms or multicopy numbers in the P. falciparum dihydrofolate reductase (pfdhfr) and P. falciparum dihydropteroate synthase (pfdhps) genes have been associated with resistance to SP [45]. Changes in the prevalence of pfcrt and pfmdr1 alleles have been observed in many areas where ACTs including amodiaquine or lumefantrine have been intensively used [46,47]. However, clinical efficacy of leading ACTs that include lumefantrine, amodiaquine, piperaquine, or mefloquine appears to remain acceptable in areas outside the GMS. Recent research suggests that plasmepsin 2–3 is associated with clinical and in vitro piperaquine resistance (PSA, piperaquine survival assay) but other markers could also be involved [48]. In Southeast Asia, intensive use of dihydroartemisinin-piperaquine (DP) in parasites already resistant to piperaquine and artemisinin has selected parasites with multiple resistance mechanisms, and high levels of treatment failure to DP are now observed in Cambodia [49].
Chloroquine remains the recommended treatment for P. vivax, but resistance and declining efficacy has been noted in several populations, and ACTs are recommended in some areas [50,51]. There are no standardized molecular correlates of chloroquine resistance for P. vivax, but P. vivax multidrug resistance 1 (pvmdr1) has been associated with resistance [52]. Beyond this, the understanding of resistance in nonfalciparum malaria is very limited.
Public health impact of resistance
While ecological studies have found broad evidence of dramatic health effects of spreading drug resistance [18], efficient assessment of the public health impact of antimalarial and insecticide resistance has been difficult. First, assessments of resistance prevalence are drawn from a few sentinel sites, but the heterogeneity of resistance in neighbouring populations can be enormous, making specific predictions difficult. Second, molecular markers are easier to measure at finer spatial and temporal scales, but the relationship with the drug or insecticide response is not direct [53,54]. Third, most policies on malaria treatment and vector control are implemented nationally, so recommending policies for regions within a country may be operationally unfeasible.
Drug resistance increases the risk of treatment failure and therefore transmission, but these relationships can be difficult to establish in the field. Human factors, especially immunity, affect treatment efficacy, so treatment failure in the whole population is not obvious until parasite resistance is well established [55]. However, in children there is a clear relationship between parasitaemia and anaemia, with associated morbidity and mortality [55,56]. Studies have correlated the prevalence of molecular markers with the risk of treatment failure, but no metric that works in all regions has been defined [57]. As a result, the prevalence of molecular markers has had a limited impact on policies for routine antimalarial use [58]. This disconnect is changing in the GMS, where ACT treatment failure has reached crisis levels [59], and rapid assessments of molecular markers for resistance to artemisinins and partner drugs are currently being used [47].
There are few published studies on the epidemiological impact of insecticide resistance, so decisions rely primarily on entomological end points. Evidence from a 5-country evaluation attempted to assess whether LLINs remain effective in the presence of pyrethroid resistance, although the studies were in areas with low to moderate resistance as measured in single-dose bioassays without assessment of resistance intensity [60]. This study was not able to quantify the effect on LLINs [59]. For IRS, the best evidence for an epidemiological impact of pyrethroid resistance comes from settings where pyrethroids were replaced in IRS campaigns with alternative insecticides and parasite prevalence rapidly declined [61, 62]. Similar evidence is available from a study in an area of Sudan with pyrethroid resistance but carbamate susceptibility, in which IRS with pyrethroids in addition to LLINs had no added impact, but changing to carbamate IRS halved the malaria incidence [60].
Managing resistance, moving toward elimination
Optimizing drug and insecticide use
Avoiding parasite or mosquito population exposure to a single selective agent is the central principle of resistance management. Ideally, insecticidal compounds with different modes of action should be used simultaneously or in spatial or temporal rotation. These principles, which are identical to those used in the management of insecticides used for crop pests, have been outlined in the GPIRM in malaria vectors [63]. Unfortunately, implementation has been challenging; pyrethroid resistance is ubiquitous, nonpyrethroid LLINs are not currently available, and other forms of vector control can significantly increase costs [64]. New public health insecticides with different modes of action are on the horizon [65], but we lack information on the effectiveness of the proposed strategies to slow the emergence or spread of insecticide resistance, and there is no clear indication of how they should be integrated alongside existing tools. This includes those that are noninsecticidal and products that work on different targets, e.g., spatial repellents and endocticides, whose efficacy may not be influenced by insecticide resistance [65]. Another confounder is the application of most insecticides for both agricultural and public health use. The impact of this on public health is highly variable depending on crop type and volume and timing of insecticide application.
What are the benefits of insecticide rotations, mixtures, or spatial mosaics of different compounds? What is the impact of adding nonpyrethroid IRS where LLINs are already deployed at high coverage and quality? When should new insecticides be adopted? What is the ideal rotation period or mosaic configuration? How many insecticide classes are needed for effective rotation or mosaic strategies? Despite the absence of data to answer these questions, some countries have already developed operational frameworks for resistance management that could be adopted by other programmes [66].
ACTs are still effective in most regions outside the GMS. Optimisation of dose, duration of treatment, timing of treatment, and pharmacokinetic-dynamic profiles in specific subpopulations, e.g., children and pregnant women, should be systematically encouraged post-licensure to maximise efficacy and slow selection for resistance. Pooled analyses have assessed the effect of dosing strategies for the several currently used ACTs, but the uptake of this by malaria control programmes is limited [67]. Molecular markers are being used in addition to therapeutic efficacy studies in specific locations in the GMS to choose treatment policies more accurately [67], but far more complete information on all ACTs is needed.
Different published models diverged on the conclusion that implementation of multiple first-line therapies could more effectively prevent the emergence of drug resistance compared with the temporal rotation or sequential use of first-line treatments [68–71]. Multiple models need to be evaluated and studies to verify this must be defined [72]. We also need to better understand why parasites do not seem to have developed resistance to quinine and factor this into future drug development efforts. In Southeast Asia, the use of triple therapies using existing antimalarials is currently being tested and could be considered in the context of multidrug-resistant malaria [73].
Assessment of the selective pressures and emergence of resistance to antimalarials is difficult with small-scale studies, but large-scale public health interventions may provide evidence. For example, studies should be undertaken in countries using different drug combinations for treatment and mass chemoprevention campaigns, such as seasonal malaria chemoprevention, mass drug administration, or intermittent preventive treatment in pregnancy (IPTp). Coordination of these interventions in the same locality may provide one way to reduce or disrupt the selection pressure exerted on a single class of compound [74].
Using data to support resistance management
Entomological data generated by countries vary in quantity and quality, and limited information flow between entomologists, programme managers, and research institutes has hindered advocacy efforts around improved resistance management. Linking entomological data to epidemiological outcomes is extremely complex [75] and by the time resistance has a demonstrable public health impact, it may be too late to intervene against it. However, South Africa [62], Zambia [76], and Equatorial Guinea [64] have resistance management plans in place. Similarly, molecular marker surveillance can inform which drug regimens are the most suitable for particular programmatic modes. This approach is now routine in some African countries [74,77] but is not universal. Drug-resistance monitoring in some countries also requires strengthening, and despite the tighter link to public health impact, the ability to respond rapidly may be lost if resistance monitoring is not well embedded. For both insecticides and drugs, defining the minimum essential data required for policy makers to manage resistance and ensuring that programs employ rigorous quality assurance in collecting and managing these data are critical.
Resistance surveillance is weak in many endemic countries. Inadequate attention and funding have been allocated to entomological monitoring and insecticide susceptibility research. Several countries in Africa have established sentinel sites for the longitudinal monitoring of insecticide resistance [5]. However, the methods, timing, and sampling are inconsistent, making meaningful inferences difficult [78]. Most of these sites use discriminating dose assays [79]. All bioassays are performed on 3–5-day-old mosquitoes under standard insectary conditions, so the effect of natural mosquito traits (e.g., age, blood-feeding status, circadian rhythm [80,81], and climatic variables [82]) on resistance is not assessed or reported [83]. Molecular species identification of mosquitoes undergoing resistance tests may also increase accuracy when compared to morphological identification. Techniques have been developed to measure the age distribution of mosquito populations in the laboratory [84], but a more precise, low-cost, field-applicable method is needed to allow malaria control programmes to evaluate the efficacy of vector control interventions is needed.
Anticipating the challenges of lower transmission
High-level use of interventions can suppress malaria transmission but also increases the risk of selection of resistance, creating new challenges at the later stages of elimination. Resistance surveillance in low transmission regions is increasingly expensive, and maintaining human and material capacity in the context of many other public health needs is crucial. The minimal criteria for the inclusion of new or existing therapeutics or insecticides in a multi-agent regimen must be defined. For drugs, these criteria might depend on transmission levels and could include pharmacokinetic-dynamic profiles, mechanisms of resistance, cross resistance, and drug–drug interactions. The corresponding parameters for insecticides of persistence/residual efficacy, mechanisms of resistance/cross resistance, or compound interactions are equally relevant. If a robust resistant phenotype can be defined, whole genome sequencing of parasites and vectors can identify regions under selection very early in the process, giving clues to associated genetic changes [85].
Market strategies and getting products to market
Single first-line antimalarial treatments or insecticide monotherapies may be cheaper in the short term, but the long-term cost-effectiveness will be compromised by increasing levels of resistance [86]. Development of normative guidance on product use within a multiyear programme of interventions is essential if short-term decision-making is to change. The selection of products may be based on a number of epidemiological, entomological, logistic, and financial variables. It is critical to develop a framework that reliably costs the long-term elimination strategies, rather than short-term ‘delivered units’, and takes into account resistance management practices. As we head toward elimination, the increased cost of keeping drugs and insecticides available for a diminishing number of cases means incentives and market strategies for keeping the pipeline of products active are paramount.
Clarity is needed on the evidence required by normative bodies to approve new products and develop treatment guidelines. New tools are likely to have a higher unit price, so clear data requirements and paths to their use require definition. Without this, programme financial constraints; uncertainties around cost-effectiveness; and delays in recommendation, production, and procurement could mean products to overcome resistance are underutilised. If this situation becomes the norm, incentives for innovation will diminish and the pipeline of efficacious tools will soon be depleted.
Conclusion
Resistance is an inevitable consequence of drug and insecticide treatment, but the malaria community as a whole has repeatedly failed to respond to this issue in a proactive way. Programme and policy decisions should be based on comprehensive resistance data, and this should be coupled with improved efforts to understand the complex biological processes that select for resistant phenotypes. The tools to surmount resistance are limited and little is known about the most effective resistance management measures, so new therapeutics and vector control products should have a clear route to market and be carefully implemented and evaluated to optimise the choice of interventions. Multidrug and insecticide regimens are not unique to malaria control and other disease systems such as HIV [87], tuberculosis [88], and agricultural pest control [89] offer important insights into the management of insecticide- and drug-based approaches. The malaria community must learn from other disease groups and industries and heed the lessons of the past or risk further erosion of the malaria elimination agenda as renewed efforts are undermined by resistance.
Zdroje
1. Bhatt S, Weiss DJ, Cameron E, Bisanzio D, Mappin B, Dalrymple U, et al. The effect of malaria control on Plasmodium falciparum in Africa between 2000 and 2015. Nature.; 2015;526: 207–211. doi: 10.1038/nature15535 26375008
2. Alonso PL, Brown G, Arevalo-Herrera M, Binka F, Chitnis C, Collins F, et al. A research agenda to underpin malaria eradication. PLoS Med. 2011;8: e1000406. doi: 10.1371/journal.pmed.1000406 21311579
3. White N, Nosten F, Looareesuwan S, Watkins W, Marsh K, Snow R, et al. Averting a malaria disaster. Lancet. 1999;353: 1965–1967. doi: 10.1016/S0140-6736(98)07367-X 10371589
4. Hemingway J, Ranson H, Magill A, Kolaczinski J, Fornadel C, Gimnig J, et al. Averting a malaria disaster: will insecticide resistance derail malaria control? Lancet. 2016; doi: 10.1016/S0140-6736(15)00417-1
5. Knox TB, Juma EO, Ochomo EO, Pates Jamet H, Ndungo L, Chege P, et al. An online tool for mapping insecticide resistance in major Anopheles vectors of human malaria parasites and review of resistance status for the Afrotropical region. Parasit Vectors. 2014;7: 76. doi: 10.1186/1756-3305-7-76 24559061
6. Dai Y, Huang X, Cheng P, Liu L, Wang H, Wang H, et al. Development of insecticide resistance in malaria vector Anopheles sinensis populations from Shandong province in China. Malar J. 2015;14: 62. doi: 10.1186/s12936-015-0592-8 25880316
7. WHO | World Malaria Report 2015. WHO. World Health Organization; 2016
8. Toé KH, Jones CM, N’Fale S, Ismail HM, Dabiré RK, Ranson H. Increased pyrethroid resistance in malaria vectors and decreased bed net effectiveness, Burkina Faso. Emerg Infect Dis. 2014;20: 1691–6. doi: 10.3201/eid2010.140619 25279965
9. Ranson H, Lissenden N. Insecticide Resistance in African Anopheles Mosquitoes: A Worsening Situation that Needs Urgent Action to Maintain Malaria Control. Trends Parasitol. 2015;32: 187–196. doi: 10.1016/j.pt.2015.11.010 26826784
10. Glunt KD, Abílio AP, Bassat Q, Bulo H, Gilbert AE, Huijben S, et al. Long-lasting insecticidal nets no longer effectively kill the highly resistant Anopheles funestus of southern Mozambique. Malar J. 2015;14: 298. doi: 10.1186/s12936-015-0807-z 26242977
11. Riveron JM, Chiumia M, Menze BD, Barnes KG, Irving H, Ibrahim SS, et al. Rise of multiple insecticide resistance in Anopheles funestus in Malawi: a major concern for malaria vector control. Malar J. 2015;14: 344. doi: 10.1186/s12936-015-0877-y 26370361
12. Hargreaves K, Koekemoer LL, Brooke BD, Hunt RH, Mthembu J, Coetzee M. Anopheles funestus resistant to pyrethroid insecticides in South Africa. Med Vet Entomol. 2000;14: 181–9. Available: http://www.ncbi.nlm.nih.gov/pubmed/10872862. Date accessed 17 Sept 2017 10872862
13. Schuurkamp GJ, Spicer PE, Kereu RK, Bulungol PK, Rieckmann KH. Chloroquine-resistant Plasmodium vivax in Papua New Guinea. Trans R Soc Trop Med Hyg. 86: 121–2. Available: http://www.ncbi.nlm.nih.gov/pubmed/1440763. Date accessed 17 Sept 2017 1440763
14. Mehlotra RK, Fujioka H, Roepe PD, Janneh O, Ursos LMB, Jacobs-Lorena V, et al. Evolution of a unique Plasmodium falciparum chloroquine-resistance phenotype in association with pfcrt polymorphism in Papua New Guinea and South America. Proc Natl Acad Sci. 2001;98: 12689–12694. doi: 10.1073/pnas.221440898 11675500
15. Wootton JC, Feng X, Ferdig MT, Cooper RA, Mu J, Baruch DI, et al. Genetic diversity and chloroquine selective sweeps in Plasmodium falciparum. Nature. 2002;418: 320–3. doi: 10.1038/nature00813 12124623
16. Ariey F, Fandeur T, Durand R, Randrianarivelojosia M, Jambou R, Legrand E, et al. Invasion of Africa by a single pfcrt allele of South East Asian type. Malar J. 2006;5: 34. doi: 10.1186/1475-2875-5-34 16638153
17. Roper C, Pearce R, Bredenkamp B, Gumede J, Drakeley C, Mosha F, et al. Antifolate antimalarial resistance in southeast Africa: a population-based analysis. Lancet. 2003;361: 1174–81. doi: 10.1016/S0140-6736(03)12951-0 12686039
18. Mita T, Venkatesan M, Ohashi J, Culleton R, Takahashi N, Tsukahara T, et al. Limited geographical origin and global spread of sulfadoxine-resistant dhps alleles in Plasmodium falciparum populations. J Infect Dis. 2011;204: 1980–8. doi: 10.1093/infdis/jir664 22021623
19. White NJ. Delaying antimalarial drug resistance with combination chemotherapy. Parassitologia. 1999;41: 301–8. Available: http://www.ncbi.nlm.nih.gov/pubmed/10697872. Date accessed 17 Sept 2017 10697872
20. Miotto O, Amato R, Ashley EA, MacInnis B, Almagro-Garcia J, Amaratunga C, et al. Genetic architecture of artemisinin-resistant Plasmodium falciparum. Nat Genet.; 2015;47: 226–34. doi: 10.1038/ng.3189 25599401
21. Takala-Harrison S, Jacob CG, Arze C, Cummings MP, Silva JC, Dondorp AM, et al. Independent emergence of artemisinin resistance mutations among Plasmodium falciparum in Southeast Asia. J Infect Dis. 2015;211: 670–9. doi: 10.1093/infdis/jiu491 25180241
22. Ménard D, Khim N, Beghain J, Adegnika AA, Shafiul-Alam M, Amodu O, et al. A Worldwide Map of Plasmodium falciparum K13-Propeller Polymorphisms. http://dx.doi.org/101056/NEJMoa1513137. Massachusetts Medical Society; 2016;
23. Rosenthal PJ, The interplay between drug resistance and fitness in malaria parasites. Mol. Microbiol 89, 1025–38 2013 doi: 10.1111/mmi.12349 23899091
24. Martinez-Torres D, Chandre F, Williamson MS, Darriet F, Bergé JB, Devonshire AL, et al. Molecular characterization of pyrethroid knockdown resistance (kdr) in the major malaria vector Anopheles gambiae s.s. Insect Mol Biol. 1998;7: 179–84. Available: http://www.ncbi.nlm.nih.gov/pubmed/9535162 Date accessed 17 Sept 2017 9535162
25. Ranson H, Jensen B, Vulule JM, Wang X, Hemingway J, Collins FH. Identification of a point mutation in the voltage-gated sodium channel gene of Kenyan Anopheles gambiae associated with resistance to DDT and pyrethroids. Insect Mol Biol. 2000;9: 491–497. doi: 10.1046/j.1365-2583.2000.00209.x 11029667
26. Strode C, Donegan S, Garner P, Enayati AA, Hemingway J. The impact of pyrethroid resistance on the efficacy of insecticide-treated bed nets against African anopheline mosquitoes: systematic review and meta-analysis. PLoS Med. 2014;11: e1001619. doi: 10.1371/journal.pmed.1001619 24642791
27. Bagi J, Grisales N, Corkill R, Morgan JC, N’Falé S, Brogdon WG, et al. When a discriminating dose assay is not enough: measuring the intensity of insecticide resistance in malaria vectors. Malar J. 2015;14: 210. doi: 10.1186/s12936-015-0721-4 25985896
28. Weetman D, Donnelly MJ. Evolution of insecticide resistance diagnostics in malaria vectors. Trans R Soc Trop Med Hyg. 2015;109: 291–3. doi: 10.1093/trstmh/trv017 25740955
29. Platt N, Kwiatkowska RM, Irving H, Diabaté A, Dabire R, Wondji CS. Target-site resistance mutations (kdr and RDL), but not metabolic resistance, negatively impact male mating competiveness in the malaria vector Anopheles gambiae. Heredity; 2015;115: 243–52. doi: 10.1038/hdy.2015.33 25899013
30. Alout H, Ndam NT, Sandeu MM, Djégbe I, Chandre F, Dabiré RK, et al. Insecticide resistance alleles affect vector competence of Anopheles gambiae s.s. for Plasmodium falciparum field isolates. PLoS ONE. 2013;8: e63849. doi: 10.1371/journal.pone.0063849 23704944
31. Gatton ML, Chitnis N, Churcher T, Donnelly MJ, Ghani AC, Godfray HCJ, et al. The importance of mosquito behavioural adaptations to malaria control in Africa. Evolution. 2013;67: 1218–30. doi: 10.1111/evo.12063 23550770
32. White LJ, Flegg JA, Phyo AP, Wiladpai-ngern JH, Bethell D, Plowe C, et al. Defining the in vivo phenotype of artemisinin-resistant falciparum malaria: a modelling approach. PLoS Med. 2015;12: e1001823. doi: 10.1371/journal.pmed.1001823 25919029
33. Witkowski B, Amaratunga C, Khim N, Sreng S, Chim P, Kim S, et al. Novel phenotypic assays for the detection of artemisinin-resistant Plasmodium falciparum malaria in Cambodia: in-vitro and ex-vivo drug-response studies. Lancet Infect Dis. 2013;13: 1043–9. doi: 10.1016/S1473-3099(13)70252-4 24035558
34. Ariey F, Witkowski B, Amaratunga C, Beghain J, Langlois A-C, Khim N, et al. A molecular marker of artemisinin-resistant Plasmodium falciparum malaria. Nature. 2014;505: 50–5. doi: 10.1038/nature12876 24352242
35. Straimer J, Gnädig NF, Witkowski B, Amaratunga C, Duru V, Ramadani AP, et al. Drug resistance. K13-propeller mutations confer artemisinin resistance in Plasmodium falciparum clinical isolates. Science. 2015;347: 428–31. doi: 10.1126/science.1260867 25502314
36. Taylor SM, Parobek CM, DeConti DK, Kayentao K, Coulibaly SO, Greenwood BM, et al. Absence of putative artemisinin resistance mutations among Plasmodium falciparum in Sub-Saharan Africa: a molecular epidemiologic study. J Infect Dis. 2015;211: 680–8. doi: 10.1093/infdis/jiu467 25180240
37. Kamau E, Campino S, Amenga-Etego L, Drury E, Ishengoma D, Johnson K, et al. K13-propeller polymorphisms in Plasmodium falciparum parasites from sub-Saharan Africa. J Infect Dis. 2014/11/05. KEMRI/United States Army Medical Research Unit-Kenya, Kisumu. Wellcome Trust Sanger Institute, Hinxton. Navrongo Health Research Centre. National Institute for Medical Research, Tanga, Tanzania. Wellcome Trust Centre for Human Genetics, University of Oxford; 2015;211: 1352–1355. doi: 10.1093/infdis/jiu608 25367300
38. World Health Organization. Status report on artemisinin and ACT resistance. 2015.
39. Chenet SM, Akinyi Okoth S, Huber CS, Chandrabose J, Lucchi NW, Talundzic E, et al. Independent Emergence of the Plasmodium falciparum Kelch Propeller Domain Mutant Allele C580Y in Guyana. J Infect Dis. 2016;213: 1472–5. doi: 10.1093/infdis/jiv752 26690347
40. Dondorp AM. New genetic markers for piperaquine resistance in Plasmodium falciparum. Lancet Infectious Diseases http://dx.doi.org/10.1016/S1473-3099(16)30212-5 Date accessed 17 Sept 2017
41. Maiga AW, Fofana B, Sagara I, Dembele D, Dara A, Traore OB, et al. No evidence of delayed parasite clearance after oral artesunate treatment of uncomplicated falciparum malaria in Mali. Am J Trop Med Hyg. 2012;87: 23–8. doi: 10.4269/ajtmh.2012.12-0058 22764287
42. Mok S, Ashley EA, Ferreira PE, Zhu L, Lin Z, Yeo T, et al. Drug resistance. Population transcriptomics of human malaria parasites reveals the mechanism of artemisinin resistance. Science. 2015;347: 431–5. doi: 10.1126/science.1260403 25502316
43. Chaorattanakawee S, Saunders DL, Sea D, Chanarat N, Yingyuen K, Sundrakes S, et al. Ex Vivo Drug Susceptibility Testing and Molecular Profiling of Clinical Plasmodium falciparum Isolates from Cambodia from 2008 to 2013 Suggest Emerging Piperaquine Resistance. Antimicrob Agents Chemother. 2015;59: 4631–43. doi: 10.1128/AAC.00366-15 26014942
44. Leang R, Taylor WRJ, Bouth DM, Song L, Tarning J, Char MC, et al. Evidence of Plasmodium falciparum Malaria Multidrug Resistance to Artemisinin and Piperaquine in Western Cambodia: Dihydroartemisinin-Piperaquine Open-Label Multicenter Clinical Assessment. Antimicrob Agents Chemother. 2015;59: 4719–26. doi: 10.1128/AAC.00835-15 26014949
45. Plowe C V, Kublin JG, Doumbo OK, Peters W, Jones A, Radloff P, et al. P. falciparum dihydrofolate reductase and dihydropteroate synthase mutations: epidemiology and role in clinical resistance to antifolates. Drug Resist Updat. 1998;1: 389–96. doi: 10.1016/s1368-7646(98)80014-9 17092820
46. Conrad MD, LeClair N, Arinaitwe E, Wanzira H, Kakuru A, Bigira V, et al. Comparative impacts over 5 years of artemisinin-based combination therapies on Plasmodium falciparum polymorphisms that modulate drug sensitivity in Ugandan children. J Infect Dis. 2014;210: 344–53. doi: 10.1093/infdis/jiu141 24610872
47. Thomsen TT, Madsen LB, Hansson HH, Tomás EVE, Charlwood D, Bygbjerg IC, et al. Rapid selection of Plasmodium falciparum chloroquine resistance transporter gene and multidrug resistance gene-1 haplotypes associated with past chloroquine and present artemether-lumefantrine use in Inhambane District, southern Mozambique. Am J Trop Med Hyg. 2013;88: 536–41. doi: 10.4269/ajtmh.12-0525 23382159
48. WHO Global Malaria Programme. Minutes of the Technical Expert Group on Drug Efficacy and Response. Available: http://www.who.int/malaria/mpac/mpac-oct2017-teg-drug-efficacy-response-session3.pdf?ua=1. Date accessed 14 Nov 2017.
49. Amaratunga C, Lim P, Suon S, Sreng S, Mao S, Sopha C, et al. Dihydroartemisinin–piperaquine resistance in Plasmodium falciparum malaria in Cambodia: a multisite prospective cohort study. Lancet Infect Dis. 2016;16: 357–365. doi: 10.1016/S1473-3099(15)00487-9 26774243
50. Phyo AP, Lwin KM, Price RN, Ashley EA, Russell B, Sriprawat K, et al. Dihydroartemisinin-piperaquine versus chloroquine in the treatment of Plasmodium vivax malaria in Thailand: a randomized controlled trial. Clin Infect Dis. 2011;53: 977–84. doi: 10.1093/cid/cir631 22002979
51. Price RN, von Seidlein L, Valecha N, Nosten F, Baird JK, White NJ. Global extent of chloroquine-resistant Plasmodium vivax: a systematic review and meta-analysis. Lancet Infect Dis. 2014;14: 982–91. doi: 10.1016/S1473-3099(14)70855-2 25213732
52. Schousboe ML, Ranjitkar S, Rajakaruna RS, Amerasinghe PH, Morales F, Pearce R, et al. Multiple Origins of Mutations in the mdr1 Gene—A Putative Marker of Chloroquine Resistance in P. vivax. PLoS Negl Trop Dis. 2015;9: e0004196. doi: 10.1371/journal.pntd.0004196 26539821
53. Venkatesan M, Gadalla NB, Stepniewska K, Dahal P, Nsanzabana C, Moriera C, et al. Polymorphisms in Plasmodium falciparum chloroquine resistance transporter and multidrug resistance 1 genes: parasite risk factors that affect treatment outcomes for P. falciparum malaria after artemether-lumefantrine and artesunate-amodiaquine. Am J Trop Med Hyg. 2014;91: 833–43. doi: 10.4269/ajtmh.14-0031 25048375
54. Briët OJ, Penny MA, Hardy D, Awolola TS, Van Bortel W, Corbel V, et al. Effects of pyrethroid resistance on the cost effectiveness of a mass distribution of long-lasting insecticidal nets: a modelling study. Malar J. 2013;12: 77. doi: 10.1186/1475-2875-12-77 23442575
55. Trape J-F. The Public Health Impact of Chloroquine Resistance in Africa. American Society of Tropical Medicine and Hygiene; 2001.
56. Douglas NM, Lampah DA, Kenangalem E, Simpson JA, Poespoprodjo JR, Sugiarto P, et al. Major burden of severe anemia from non-falciparum malaria species in Southern Papua: a hospital-based surveillance study. PLoS Med. 2013;10: e1001575; discussion e1001575. doi: 10.1371/journal.pmed.1001575 24358031
57. Djimde AA, Barger B, Kone A, Beavogui AH, Tekete M, Fofana B, et al. A molecular map of chloroquine resistance in Mali. FEMS Immunol Med Microbiol. 2010;58: 113–8. doi: 10.1111/j.1574-695X.2009.00641.x 20041947
58. Djimdé AA, Dolo A, Ouattara A, Diakité S, Plowe CV, Doumbo OK. Molecular Diagnosis of Resistance to Antimalarial Drugs during Epidemics and in War Zones. J Infect Dis. 2004;190: 853–855. doi: 10.1086/422758 15272415
59. Lim P, Dek D, Try V, Sreng S, Suon S, Fairhurst RM. Decreasing pfmdr1 copy number suggests that Plasmodium falciparum in Western Cambodia is regaining in vitro susceptibility to mefloquine. Antimicrob Agents Chemother. 2015;59: 2934–7. doi: 10.1128/AAC.05163-14 25712365
60. Hmooda Toto K, Bashir AI, Mnzava AP, Abdin MSE, Eltaher JS, Banaga AO, et al. The impact of insecticide resistance in Anopheles arabiensis on malaria incidence and prevalence in Sudan and the costs of mitigation. Proc. Natl Acad. Sci. In press.
61. Philip Ricks. The Importance of Insecticide Resistance Monitoring to Maintain IRS Program Effectiveness. 2015. Available: http://www.africairs.net/2015/11/the-pmi-airs-project-presents-at-astmh/ Date accessed 17 Sept 2017
62. Coetzee M, Kruger P, Hunt RH, Durrheim DN, Urbach J, Hansford CF. Malaria in South Africa: 110 years of learning to control the disease. S Afr Med J. 2013;103: 770–8. Available: http://www.ncbi.nlm.nih.gov/pubmed/24079632 doi: 10.7196/samj.7446 24079632
63. WHO | Global plan for insecticide resistance management in malaria vectors. WHO. World Health Organization; 2015
64. Hemingway J, Vontas J, Poupardin R, Raman J, Lines J, Schwabe C, et al. Country-level operational implementation of the Global Plan for Insecticide Resistance Management. Proc Natl Acad Sci U S A. 2013;110: 9397–402. doi: 10.1073/pnas.1307656110 23696658
65. Hemingway J, Beaty BJ, Rowland M, Scott TW, Sharp BL. The Innovative Vector Control Consortium: improved control of mosquito-borne diseases. Trends Parasitol. 2006;22: 308–12. doi: 10.1016/j.pt.2006.05.003 16713358
66. Chanda E, Thomsen EK, Musapa M, Kamuliwo M, Brogdon WG, Norris DE, et al. An Operational Framework for Insecticide Resistance Management Planning. Emerg Infect Dis. Centers for Disease Control and Prevention; 2016;22: 773–9. doi: 10.3201/eid2205.150984 27089119
67. Group TWARN (WWARN) DS. The effect of dosing regimens on the antimalarial efficacy of dihydroartemisinin-piperaquine: a pooled analysis of individual patient data. PLoS Med. 2013;10: e1001564; discussion e1001564. doi: 10.1371/journal.pmed.1001564 24311989
68. Boni MF, Smith DL, Laxminarayan R. Benefits of using multiple first-line therapies against malaria. Proc Natl Acad Sci. 2008;105: 14216–14221. doi: 10.1073/pnas.0804628105 18780786
69. Smith DL, Klein EY, McKenzie FE, Laxminarayan R. Prospective strategies to delay the evolution of anti-malarial drug resistance: weighing the uncertainty. Malar J. B 2010;9: 217. doi: 10.1186/1475-2875-9-217 20653960
70. Nguyen TD, Olliaro P, Dondorp AM, Baird JK, Lam HM, Farrar J, et al. Optimum population-level use of artemisinin combination therapies: a modelling study. Lancet Glob Heal. 2015;3: e758–e766. doi: 10.1016/S2214-109X(15)00162-X 26545449
71. Pongtavornpinyo W1, Yeung S, Hastings IM, Dondorp AM, Day NP, White NJ. Spread of anti-malarial drug resistance: mathematical model with implications for ACT drug policies. Malar J. 2008 Nov 2;7:229. doi: 10.1186/1475-2875-7-229 18976503
72. Boni MF, White NJ, Baird JK, Peters W, Dondorp A, Nosten F, et al. The Community As the Patient in Malaria-Endemic Areas: Preempting Drug Resistance with Multiple First-Line Therapies. PLoS Med. 2016;13: e1001984. doi: 10.1371/journal.pmed.1001984 27022739
73. Maxmen A. Back on TRAC: New trial launched in bid to outpace multidrug-resistant malaria. Nat Med. Nature Research; 2016;22: 220–221. doi: 10.1038/nm0316-220 26937610
74. Hamma M. Impact of seasonal malaria chemoprevention of sulphadoxinepyrimethamine plus amodiaquine on molecular markers resistance of Plasmodium falciparum malaria: A review in West Africa. Clin Rev Opin. A2016;7: 1–10. doi: 10.5897/CRO15.0098
75. Kleinschmidt I, Mnzava AP, Kafy HT, Mbogo C, Bashir AI, Bigoga J, et al. Design of a study to determine the impact of insecticide resistance on malaria vector control: a multi-country investigation. Malar J. 2015;14: 282. doi: 10.1186/s12936-015-0782-4 26194648
76. Thomsen EK, Strode C, Hemmings K, Hughes AJ, Chanda E, Musapa M, et al. Underpinning sustainable vector control through informed insecticide resistance management. PLoS ONE. 2014;9: e99822. doi: 10.1371/journal.pone.0099822 24932861
77. Mugittu K, Ndejembi M, Malisa A, Lemnge M, Premji Z, Mwita A, et al. Therapeutic efficacy of sulfadoxine-pyrimethamine and prevalence of resistance markers in Tanzania prior to revision of malaria treatment policy: Plasmodium falciparum dihydrofolate reductase and dihydropteroate synthase mutations in monitoring in vivo re. Am J Trop Med Hyg. 2004;71: 696–702. Available: http://www.ncbi.nlm.nih.gov/pubmed/15642957 15642957
78. Ranson H, Lissenden N. Insecticide Resistance in African Anopheles Mosquitoes: A Worsening Situation that Needs Urgent Action to Maintain Malaria Control. Trends Parasitol. 2015;32: 187–196. doi: 10.1016/j.pt.2015.11.010 26826784
79. Bagi J, Grisales N, Corkill R, Morgan JC, N’Falé S, Brogdon WG, et al. When a discriminating dose assay is not enough: measuring the intensity of insecticide resistance in malaria vectors. Malar J. 2015;14: 210. doi: 10.1186/s12936-015-0721-4 25985896
80. Kulma K, Saddler A, Koella JC. Effects of age and larval nutrition on phenotypic expression of insecticide-resistance in Anopheles mosquitoes. PLoS ONE. 2013;8: e58322. doi: 10.1371/journal.pone.0058322 23484017
81. Oliver S V, Brooke BD, Sinka M, Bangs M, Manguin S, Coetzee M, et al. The effect of multiple blood-feeding on the longevity and insecticide resistant phenotype in the major malaria vector Anopheles arabiensis (Diptera: Culicidae). Parasit Vectors. 2014;7: 390. doi: 10.1186/1756-3305-7-390 25150975
82. Glunt KD, Paaijmans KP, Read AF, Thomas MB, Maxmen A, Kelly-Hope L, et al. Environmental temperatures significantly change the impact of insecticides measured using WHOPES protocols. Malar J. 2014;13: 350. doi: 10.1186/1475-2875-13-350 25187231
83. Strode C, Steen K, Ortelli F, Ranson H. Differential expression of the detoxification genes in the different life stages of the malaria vector Anopheles gambiae. Insect Mol Biol. 2006;15: 523–30. doi: 10.1111/j.1365-2583.2006.00667.x 16907839
84. Sikulu MT, Majambere S, Khatib BO, Ali AS, Hugo LE, Dowell FE. Using a near-infrared spectrometer to estimate the age of anopheles mosquitoes exposed to pyrethroids. PLoS ONE. 2014;9: e90657. doi: 10.1371/journal.pone.0090657 24594705
85. Donnelly MJ, Isaacs AT, Weetman D. Identification, Validation, and Application of Molecular Diagnostics for Insecticide Resistance in Malaria Vectors. Trends Parasitol. 2015;32: 197–206. doi: 10.1016/j.pt.2015.12.001 26750864
86. Lubell Y, Dondorp A, Guérin PJ, Drake T, Meek S, Ashley E, et al. Artemisinin resistance—modelling the potential human and economic costs. Malar J. 2014;13: 452. doi: 10.1186/1475-2875-13-452 25418416
87. Volberding PA, Deeks SG. Antiretroviral therapy and management of HIV infection. Lancet. 2010;376: 49–62. doi: 10.1016/S0140-6736(10)60676-9 20609987
88. Goldberg DE, Siliciano RF, Jacobs WR. Outwitting evolution: fighting drug-resistant TB, malaria, and HIV. Cell. 2012;148: 1271–83. doi: 10.1016/j.cell.2012.02.021 22424234
89. Mitchell PD, Onstad DW. Chapter 2 –Valuing Pest Susceptibility to Control. Insect Resistance Management. 2014. pp. 25–53. doi: 10.1016/B978-0-12-396955-2.00002–3
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