BackgroundMalaria vectors have acquired widespread resistance throughout sub-Saharan Africa to many of the currently used insecticides. Hence, there is an urgent need to develop alternative strategies including the development of new insecticides for effective management of insecticide resistance. To maintain progress against malaria, it is necessary to identify other residual insecticides for mosquito nets. In the present WHOPES phase II analogue study, the utility of chlorfenapyr, a pyrrole class insecticide mixed with alpha-cypermethrin on a long-lasting mosquito bed net was evaluated against Anopheles gambiae s.l.MethodsBed nets treated with chlorfenapyr and alpha-cypermethrin and mixture of both compounds were tested for their efficacy on mosquitoes. Washed (20 times) and unwashed of each type of treated nets and were tested according to WHOPES guidelines. Efficacy of nets were expressed in terms of blood-feeding inhibition rate, deterrence, induced exophily and mortality rate. The evaluation was conducted in experimental huts of Vallée du Kou seven (VK7) in Burkina Faso (West Africa) following WHOPES phase II guidelines. In addition, a WHOPES phase I evaluation was also performed.ResultsMixture treated nets killed significantly (P < 0.05) more mosquitoes than solo alpha-cypermethrin nets, unwashed and washed. Proportionally, this equated to mortalities of 78 and 76% (for mixture nets) compared to only 17 and 10% (for solo alpha-cypermethrin) to An. gambiae, respectively. In contrast mixture net proportions were not significantly (P > 0.05) different from nets treated with chlorfenapyr 200 mg/m2 unwashed (86%). The washed and unwashed nets treated with the mixtures resulted in personal protection against An. gambiae s.l. biting 34 and 44%. In contrast the personal protection observed for washed and unwashed alpha-cypermethrin treated nets generated (14 and 24%), and chlorfenapyr solo treated net was rather low (22%).ConclusionAmong all nets trialled, the combination of chlorfenapyr and alpha-cypermethrin on bed nets provided better mortality in phase II after 20 washes. Results suggest that this combination could be a potential insecticide resistance management tool for preventing malaria transmission in areas compromised by the spread of pyrethroid resistance.

Adult females An. gambiae were exposed in 3 min cone test to treated nets with PPF before or after they were blood fed. The effects of PPF on ovaries development, females oviposition and eggs hatching were assessed. Both unfed and fed mosquitoes exposed to PPF exhibited nearly complete inhibition of fecundity (70-100%) and fertility (90-100%). After females have been exposed once to PPF, the sterilizing effect on their fecundity was observed over 3 consecutive blood meals suggesting that PPF might have an irreversible sterilizing effect. Observation of the ovaries of exposed females to PPF under microscope revealed that the ovaries failed to develop even after several blood meals. The combination of PPF to pyrethroids on bednets could provide better malaria control tool and prevent the further development and spread of pyrethroid resistance in malaria vectors.

Background The continued spread of insecticide resistance in mosquito vectors of malaria and arboviral diseases may lead to operational failure of insecticide-based interventions if resistance is not monitored and managed efficiently. This study aimed to develop and validate a new WHO glass bottle bioassay method as an alternative to the WHO standard insecticide tube test to monitor mosquito susceptibility to new public health insecticides with particular modes of action, physical properties or both. Methods A multi-centre study involving 21 laboratories worldwide generated data on the susceptibility of seven mosquito species (Aedes aegypti, Aedes albopictus, Anopheles gambiae sensu stricto [An. gambiae s.s.], Anopheles funestus, Anopheles stephensi, Anopheles minimus and Anopheles albimanus) to seven public health insecticides in five classes, including pyrethroids (metofluthrin, prallethrin and transfluthrin), neonicotinoids (clothianidin), pyrroles (chlorfenapyr), juvenile hormone mimics (pyriproxyfen) and butenolides (flupyradifurone), in glass bottle assays. The data were analysed using a Bayesian binomial model to determine the concentration–response curves for each insecticide–species combination and to assess the within-bioassay variability in the susceptibility endpoints, namely the concentration that kills 50% and 99% of the test population (LC50 and LC99, respectively) and the concentration that inhibits oviposition of the test population by 50% and 99% (OI50 and OI99), to measure mortality and the sterilizing effect, respectively. Results Overall, about 200,000 mosquitoes were tested with the new bottle bioassay, and LC50/LC99 or OI50/OI99 values were determined for all insecticides. Variation was seen between laboratories in estimates for some mosquito species–insecticide combinations, while other test results were consistent. The variation was generally greater with transfluthrin and flupyradifurone than with the other compounds tested, especially against Anopheles species. Overall, the mean within-bioassay variability in mortality and oviposition inhibition were < 10% for most mosquito species-insecticide combinations. Conclusion Our findings, based on the largest susceptibility dataset ever produced on mosquitoes, showed that the new WHO bottle bioassay is adequate for evaluating mosquito susceptibility to new and promising public health insecticides currently deployed for vector control. The datasets presented in this study have been used recently by the WHO to establish 17 new insecticide discriminating concentrations (DCs) for either Aedes spp. or Anopheles spp. The bottle bioassay and DCs can now be widely used to monitor baseline insecticide susceptibility of wild populations of vectors of malaria and Aedes-borne diseases worldwide. Graphical abstract