Transmission of Plasmodium falciparum malaria parasites occurs when nocturnal Anopheles mosquito vectors feed on human blood. In Africa, where malaria burden is greatest, bednets treated with pyrethroid insecticide were highly effective in preventing mosquito bites and reducing transmission, and essential to achieving unprecedented reductions in malaria until 2015 1. Since then, progress has stalled 2 and with insecticidal bednets losing efficacy against pyrethroid-resistant Anopheles vectors 3,4 , methods that restore performance are urgently needed to eliminate any risk of malaria returning to the levels seen prior to their widespread use throughout sub-Saharan Africa 5. Here we show that the primary malaria vector Anopheles gambiae is targeted and killed by small insecticidal net barriers positioned above a standard bednet, in a spatial region of high mosquito activity but zero contact with sleepers, opening the way for deploying many more insecticides on bednets than currently possible. Tested against wild pyrethroid-resistant Anopheles gambiae in Burkina Faso, pyrethroid bednets with organophosphate barriers achieved significantly higher killing rates than bednets alone. Treated barriers on untreated bednets were equally effective, without significant loss of personal protection. Mathematical modelling of transmission dynamics predicted reductions in clinical malaria incidence with barrier bednets that matched those of 'next-generation' nets recommended by WHO against resistant vectors. Mathematical
26The functional response describes the relationship between feeding rate and prey density, and 27 is important ecologically as it describes how the foraging behaviour of an animal changes in 28 response to food availability. The effects of habitat complexity and food item size was
The functional response is the relationship between the feeding rate of an animal and its food density. It is reliant on two basic parameters; the volume searched for prey per unit time (searching rate) and the time taken to consume each prey item (handling time). As fish functional responses can be difficult to determine directly, it may be more feasible to measure their underlying behavioural parameters in controlled conditions and use these to predict the functional response. Here, we tested how accurately a Type II functional response model predicted the observed functional response of roach Rutilus rutilus, a visually foraging fish, and compared it with Type I functional response. Foraging experiments were performed by exposing fish in tank aquaria to a range of food densities, with their response captured using a two-camera videography system. This system was validated and was able to accurately measure fish behaviour in the aquaria, and enabled estimates of fish reaction distance, swimming speed (from which searching rate was calculated) and handling time to be measured. The parameterised Type II functional response model accurately predicted the observed functional response and was superior to the Type I model. These outputs suggest it will be possible to accurately measure behavioural parameters in other animal species and use these to predict the functional response in situations where it cannot be observed directly.
Robust imaging techniques for tracking insects have been essential tools in numerous laboratory and field studies on pests, beneficial insects and model systems. Recent innovations in optical imaging systems and associated signal processing have enabled detailed characterization of nocturnal mosquito behaviour around bednets and improvements in bednet design, a global essential for protecting populations against malaria. Nonetheless, there remain challenges around ease of use for large-scale in situ recordings and extracting data reliably in the critical areas of the bednet where the optical signal is attenuated. Here, we introduce a retro-reflective screen at the back of the measurement volume, which can simultaneously provide diffuse illumination, and remove optical alignment issues while requiring only one-sided access to the measurement space. The illumination becomes significantly more uniform, although noise removal algorithms are needed to reduce the effects of shot noise, particularly across low-intensity bednet regions. By systematically introducing mosquitoes in front of and behind the bednet in laboratory experiments, we are able to demonstrate robust tracking in these challenging areas. Overall, the retro-reflective imaging set-up delivers mosquito segmentation rates in excess of 90% compared to less than 70% with backlit systems.
Background The WHO cone test is one of three tests currently used to evaluate the efficacy of insecticide-treated bed nets (ITNs). It generates two test outputs, knockdown and 24-h mortality, both indicative of immediate toxicity but that reveal little about the nature of mosquito and ITN interaction or how results translate to real-world settings. Methods A human arm held 5 mm behind the net surface acted as a host attractant during cone tests and a smartphone was used to capture mosquito behaviour in the cone. Post-exposure blood feeding and survival for nine days were recorded; ingested blood meal size was determined by measuring excreted haematin. Four strains of Anopheles gambiae (insecticide susceptible: Kisumu and N’gousso; insecticide resistant: Banfora and VK7) were tested with and without the host attractant using untreated, Permanet 2.0 and Olyset nets. Video recordings were scan sampled every five seconds to record mosquito positions on either the net, in flight or in contact with the cone. Generalized estimating equations were used to analyse all data except survival within nine days which was analysed using Weighted Cox Regression. Results Net contact was the most frequently recorded behaviour in all Anopheles spp. strains on all nets. Adding the human host as attractant triggered excitatory behaviours: in all strains, the magnitude of net contact was significantly decreased compared to tests without a host. ITN exposure altered the observed behaviour of the two susceptible strains, which exhibited a decreased response to the host during ITN tests. The resistant strains did not alter their behaviour during ITN tests. Significantly less net contact was observed during Olyset Net tests compared to Permanet 2.0. The host presence affected survival after exposure: Banfora and VK7 mosquitoes exposed to Permanet 2.0 with a host lived longer compared to tests performed without a host. However, mosquitoes that blood-fed and survived long enough to digest the blood meal did not exhibit significantly reduced longevity regardless of the presence of the host attractant. Conclusions Simple modifications to the WHO cone test and extension of post-test monitoring beyond the current 24 h enable detailed behavioural characterizations of individual ITNs to be compiled. The effects observed from testing with a host and including blood feeding suggest that more representative estimates of true of ITN efficacy are gained with these modifications than when using the current testing protocol.
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