Long-Lasting Insecticidal Nets Incorporating Piperonyl Butoxide Reduce the Risk of Malaria in Children in Western Kenya: A Cluster Randomized Controlled Trial.

Malaria vectors have acquired an enzyme that metabolizes pyrethroids. To tackle this problem, we evaluated long-lasting insecticidal nets incorporating piperonyl butoxide (PBO-LLINs) with a community-based cluster randomized control trial in western Kenya. The primary endpoints were anopheline density and Plasmodium falciparum polymerase chain reaction (PCR)-positive prevalence (PCRpfPR) of children aged 7 months to 10 years. Four clusters were randomly selected for each of the treatment and control arms (eight clusters in total) from 12 clusters, and PBO-LLINs and standard LLINs were distributed in February 2011 to 982 and 1,028 houses for treatment and control arms, respectively. Entomological surveys targeted 20 houses in each cluster, and epidemiological surveys targeted 150 children. Cluster-level permutation tests evaluated the effectiveness using the fitted values from individual level regression models adjusted for baseline. Bootstrapping estimated 95% confidence intervals (CIs). The medians of anophelines per house were 1.4 (interquartile range [IQR]: 2.3) and 3.4 (IQR: 3.7) in the intervention and control arms after 3 months, and 0.4 (IQR: 0.2) and 1.6 (IQR: 0.5) after 10 months, respectively. The differences were -2.5 (95% CI: -6.4 to -0.6) and -1.3 (95% CI: -2.0 to -0.7), respectively. The datasets of 861 and 775 children were analyzed in two epidemiological surveys. The median PCRpfPRs were 25% (IQR: 11%) in the intervention arm and 52% (IQR: 11%) in the control arm after 5 months and 33% (IQR: 11%) and 45% (IQR: 5%) after 12 months. The PCRpfPR ratios were 0.67 (95% CI: 0.38, 0.91) and 0.74 (95% CI: 0.53, 0.90), respectively. We confirmed the superiority of PBO-LLINs.

An Experimental Hut Evaluation of PBO-Based and Pyrethroid-Only Nets against the Malaria Vector Anopheles funestus Reveals a Loss of Bed Nets Efficacy Associated with GSTe2 Metabolic Resistance.

Growing insecticide resistance in malaria vectors is threatening the effectiveness of insecticide-based interventions, including Long Lasting Insecticidal Nets (LLINs). However, the impact of metabolic resistance on the effectiveness of these tools remains poorly characterized. Using experimental hut trials and genotyping of a glutathione S-transferase resistance marker (L119F-GSTe2), we established that GST-mediated resistance is reducing the efficacy of LLINs against Anopheles funestus. Hut trials performed in Cameroon revealed that Piperonyl butoxide (PBO)-based nets induced a significantly higher mortality against pyrethroid resistant An. funestus than pyrethroid-only nets. Blood feeding rate and deterrence were significantly higher in all LLINs than control. Genotyping the L119F-GSTe2 mutation revealed that, for permethrin-based nets, 119F-GSTe2 resistant mosquitoes have a greater ability to blood feed than susceptible while the opposite effect is observed for deltamethrin-based nets. For Olyset Plus, a significant association with exophily was observed in resistant mosquitoes (OR = 11.7; p < 0.01). Furthermore, GSTe2-resistant mosquitoes (cone assays) significantly survived with PermaNet 2.0 (OR = 2.1; p < 0.01) and PermaNet 3.0 (side) (OR = 30.1; p < 0.001) but not for Olyset Plus. This study shows that the efficacy of PBO-based nets (e.g., blood feeding inhibition) against pyrethroid resistant malaria vectors could be impacted by other mechanisms including GST-mediated metabolic resistance not affected by the synergistic action of PBO. Mosaic LLINs incorporating a GST inhibitor (diethyl maleate) could help improve their efficacy in areas of GST-mediated resistance.

Anopheles gambiae populations from Burkina Faso show minimal delayed mortality after exposure to insecticide-treated nets.

BACKGROUND: The efficacy of long-lasting insecticidal nets (LLINs) in preventing malaria in Africa is threatened by insecticide resistance. Bioassays assessing 24-hour mortality post-LLIN exposure have established that resistance to the concentration of pyrethroids used in LLINs is widespread. However, although mosquitoes may no longer be rapidly killed by LLIN exposure, a delayed mortality effect has been shown to reduce the transmission potential of mosquitoes exposed to nets. This has been postulated to partially explain the continued efficacy of LLINs against pyrethroid-resistant populations. Burkina Faso is one of a number of countries with very high malaria burdens and pyrethroid-resistant vectors, where progress in controlling this disease has stagnated. We measured the impact of LLIN exposure on mosquito longevity in an area of the country with intense pyrethroid resistance to establish whether pyrethroid exposure was still shortening mosquito lifespan in this setting. METHODS: We quantified the immediate and delayed mortality effects of LLIN exposure using standard laboratory WHO cone tests, tube bioassays and experimental hut trials on Anopheles gambiae populations originating from the Cascades region of Burkina Faso using survival analysis and a Bayesian state-space model. RESULTS: Following single and multiple exposures to a PermaNet 2.0 LLIN only one of the four mosquito populations tested showed evidence of delayed mortality. No delayed mortality was seen in experimental hut studies using LLINs. A delayed mortality effect was only observed in WHO tube bioassays when deltamethrin concentration was increased above the standard diagnostic dose. CONCLUSIONS: As mosquito pyrethroid-resistance increases in intensity, delayed effects from LLIN exposure are substantially reduced or absent. Given the rapid increase in resistance occurring in malaria vectors across Africa it is important to determine whether the failure of LLINs to shorten mosquito lifespan is now a widespread phenomenon as this will have important implications for the future of this pivotal malaria control tool.

Escalation of Pyrethroid Resistance in the Malaria Vector Anopheles funestus Induces a Loss of Efficacy of Piperonyl Butoxide-Based Insecticide-Treated Nets in Mozambique.

BACKGROUND: Insecticide resistance poses a serious threat to insecticide-based interventions in Africa. There is a fear that resistance escalation could jeopardize malaria control efforts. Monitoring of cases of aggravation of resistance intensity and its impact on the efficacy of control tools is crucial to predict consequences of resistance. METHODS: The resistance levels of an Anopheles funestus population from Palmeira, southern Mozambique, were characterized and their impact on the efficacy of various insecticide-treated nets established. RESULTS: A dramatic loss of efficacy of all long-lasting insecticidal nets (LLINs), including piperonyl butoxide (PBO)-based nets (Olyset Plus), was observed. This An. funestus population consistently (2016, 2017, and 2018) exhibited a high degree of pyrethroid resistance. Molecular analyses revealed that this resistance escalation was associated with a massive overexpression of the duplicated cytochrome P450 genes CYP6P9a and CYP6P9b, and also the fixation of the resistance CYP6P9a_R allele in this population in 2016 (100%) in contrast to 2002 (5%). However, the low recovery of susceptibility after PBO synergist assay suggests that other resistance mechanisms could be involved. CONCLUSIONS: The loss of efficacy of pyrethroid-based LLINs with and without PBO is a concern for the effectiveness of insecticide-based interventions, and action should be taken to prevent the spread of such super-resistance.

Nets versus spraying: A spatial modelling approach reveals indoor residual spraying targets Anopheles mosquito habitats better than mosquito nets in Tanzania.

The global implementation of malaria interventions has averted hundreds of millions of clinical malaria cases in the last decade. This study assesses predicted Anopheles mosquito distributions across the United Republic of Tanzania before large-scale insecticide-treated net (ITN) rollouts and indoor residual spraying (IRS) initiatives to determine whether mosquito net usage by children under the age of five and IRS are targeted to areas where historical evidence indicates mosquitoes thrive. Demographic and Health Surveys data from 2011-2012 and 2015-2016 include detailed measurements of mosquito net and IRS use across Tanzania. Anopheline data are far less intensively collected, but we constructed a Maxent-built baseline mosquito habitat suitability (MHS) map (AUC = 0.872) with Tanzanian Anopheles occurrence records from 1999-2003. This MHS model was tested against independently-observed georeferenced Plasmodium falciparum cases from the Malaria Atlas Project, with ~87% of cases from 1999-2003 (n = 107) and ~84% of cases from 1985-2012 (n = 919) occurring in areas of high predicted suitability for mosquitoes. We compared the validated MHS with subsequent malaria interventions using mixed effects logistic regression. Specifically, we assessed whether Anopheles habitat suitability related to the frequency that ≥1 child in a household reportedly slept under a mosquito net when that intervention later became widely available, and whether IRS was reportedly applied to dwellings over a one-year period. There was no evidence that mosquito net use the night before the survey related to MHS from 2011-2012 and marginally significant evidence (p<0.05) from 2015-2016 (ß = 1.466, 95% C.I. = 0.848-2.103, marginal R2 = 0.020, respectively). However, the likelihood of IRS treatments rose relatively strongly in the 12 months prior to both surveys (ß = 13.466, 95% C.I. = 10.488-16.456, marginal R2 = 0.144, and ß = 6.817, 95% C.I. = 5.439-8.303, marginal R2 = 0.136, respectively). IRS treatments have therefore been targeted more effectively than mosquito nets toward areas where anopheline habitat suitability was previously found to be high.

Mesocosm experiments reveal the impact of mosquito control measures on malaria vector life history and population dynamics.

The impact of control measures on mosquito vector fitness and demography is usually estimated from bioassays or indirect variables in the field. Whilst indicative, neither approach is sufficient to quantify the potentially complex response of mosquito populations to combined interventions. Here, large replicated mesocosms were used to measure the population-level response of the malaria vector Anopheles arabiensis to long-lasting insecticidal nets (LLINs) when used in isolation, or combined with insecticidal eave louvers (EL), or treatment of cattle with the endectocide Ivermectin (IM). State-space models (SSM) were fit to these experimental data, revealing that LLIN introduction reduced adult mosquito survival by 91% but allowed population persistence. ELs provided no additional benefit, but IM reduced mosquito fecundity by 59% and nearly eliminated all populations when combined with LLINs. This highlights the value of IM for integrated vector control, and mesocosm population experiments combined with SSM for identifying optimal combinations for vector population elimination.

A periodically-forced mathematical model for the seasonal dynamics of malaria in mosquitoes.

We describe and analyze a periodically-forced difference equation model for malaria in mosquitoes that captures the effects of seasonality and allows the mosquitoes to feed on a heterogeneous population of hosts. We numerically show the existence of a unique globally asymptotically stable periodic orbit and calculate periodic orbits of field-measurable quantities that measure malaria transmission. We integrate this model with an individual-based stochastic simulation model for malaria in humans to compare the effects of insecticide-treated nets (ITNs) and indoor residual spraying (IRS) in reducing malaria transmission, prevalence, and incidence. We show that ITNs are more effective than IRS in reducing transmission and prevalence though IRS would achieve its maximal effects within 2 years while ITNs would need two mass distribution campaigns over several years to do so. Furthermore, the combination of both interventions is more effective than either intervention alone. However, although these interventions reduce transmission and prevalence, they can lead to increased clinical malaria; and all three malaria indicators return to preintervention levels within 3 years after the interventions are withdrawn.

Experimental hut evaluation of Fendona 6SC®-treated bednets and Interceptor® long-lasting nets against Anopheles gambiae s.l. in Burkina Faso.

BACKGROUND & OBJECTIVES: Malaria prevention relies heavily on insecticide-treated bednets. Even though the benefits of bednets have been proven that in most of the studies carried out in Africa, their efficacy remains dependent on local conditions. In this study, under field conditions, we evaluated the efficacy of two LLINs (PermaNet® vs Interceptor® ) and two bednet treatment kits (K-O TAB® vs Fendona 6SC® ) against Anopheles gambiae s.l. METHODS: Bednets were evaluated using experimental huts in the village of Pissy located in the Saponé health district of Burkina Faso. Treatments and sleepers were randomly rotated between huts. Results are expressed in terms of induced exophily, mortality after 24 h and blood-feeding inhibition. RESULTS: A total of 1392 An. gambiae s.l. mosquitoes were collected during 120 nights in the experimental huts. The overall mortality rates were 85.4% (CL: 79.7-91.4) and 77.5% (CL: 56.9-97.3) for PermaNet® and Interceptor® ,respectively. For the conventionally treated bednets, the mortality was 78.2% (CL: 63.13-96.7) with the Fendona 6SC®-treated nets and 75.5% (CL: 61.2-93) with the K-O TAB®-treated nets. The proportion of blood-fed mosquitoes was significantly higher in the untreated bednet arm than in the treated one, as well as for long-lasting nets than for conventionally treated nets. The entry rate did not vary significantly according to the bednet type, but the treated bednets increased the level of exophily by at least 43%. CONCLUSION: In the field, the Fendona 6SC® kit and the Interceptor bednets showed comparable efficacy to the already used K-O TAB® kit and PermaNet® bednets. These results could help National Malaria Program managers to formulate appropriate policy for effective vector control.