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Vector control is a key means of combating mosquito-borne diseases and the only tool available for tackling the transmission of dengue, a disease for which no vaccine, prophylaxis, or therapeutant currently exists. The most effective mosquito control methods include a variety of insecticidal tools that target adults or juveniles. Their successful implementation depends on impacting the largest proportion of the vector population possible. We demonstrate a control strategy that dramatically improves the efficiency with which high coverage of aquatic mosquito habitats can be achieved. The method exploits adult mosquitoes as vehicles of insecticide transfer by harnessing their fundamental behaviors to disseminate a juvenile hormone analogue (JHA) between resting and oviposition sites. A series of field trials undertaken in an Amazon city (Iquitos, Peru) showed that the placement of JHA dissemination stations in just 3–5% of the available resting area resulted in almost complete coverage of sentinel aquatic habitats. More than control mortality occurred in 95–100% of the larval cohorts of Aedes aegypti developing at those sites. Overall reductions in adult emergence of 42–98% were achieved during the trials. A deterministic simulation model predicts amplifications in coverage consistent with our observations and highlights the importance of the residual activity of the insecticide for this technique.

Mosquito-borne viruses threaten public health worldwide. When the ratio of competent vectors to susceptible humans is low enough, the virus's basic reproductive number (R0) falls below 1.0 (each case generating, on average, <1.0 additional case) and the infection fades out from the population. Conventional mosquito control tactics, however, seldom yield R0 < 1.0. A promising alternative uses mosquitoes to disseminate a potent growth-regulator larvicide, pyriproxyfen (PPF), to aquatic larval habitats; this kills most mosquito juveniles and substantially reduces adult mosquito emergence. We tested mosquito-disseminated PPF in Manacapuru, a 60,000-inhabitant city (~650 ha) in Amazonian Brazil. We sampled juvenile mosquitoes monthly in 100 dwellings over four periods in February 2014-January 2016: 12 baseline months, 5 mo of citywide PPF dissemination, 3 mo of focal PPF dissemination around Aedes-infested dwellings, and 3 mo after dissemination ended. We caught 19,434 juvenile mosquitoes (66% Aedes albopictus, 28% Ae. aegypti) in 8,271 trap-months. Using generalized linear mixed models, we estimated intervention effects on juvenile catch and adult emergence while adjusting for dwelling-level clustering, unequal sampling effort, and weather-related confounders. Following PPF dissemination, Aedes juvenile catch decreased by 79%-92% and juvenile mortality increased from 2%-7% to 80%-90%. Mean adult Aedes emergence fell from 1,077 per month (range 653-1,635) at baseline to 50.4 per month during PPF dissemination (range 2-117). Female Aedes emergence dropped by 96%-98%, such that the number of females emerging per person decreased to 0.06 females per person-month (range 0.002-0.129). Deterministic models predict, under plausible biological-epidemiological scenarios, that the R0 of typical Aedes-borne viruses would fall from 3-45 at baseline to 0.004-0.06 during PPF dissemination. The main limitations of our study were that it was a before-after trial lacking truly independent replicates and that we did not measure mosquito-borne virus transmission empirically. Mosquito-disseminated PPF has potential to block mosquito-borne virus transmission citywide, even under adverse scenarios. Our results signal new avenues for mosquito-borne disease prevention, likely including the effective control of Aedes-borne dengue, Zika, and chikungunya epidemics. Cluster-randomized controlled trials will help determine whether mosquito-disseminated PPF can, as our findings suggest, develop into a major tool for improving global public health.

Mosquito-borne pathogens pose major public health challenges worldwide. With vaccines or effective drugs still unavailable for most such pathogens, disease prevention heavily relies on vector control. To date, however, mosquito control has proven difficult, with low breeding-site coverage during control campaigns identified as a major drawback. A novel tactic exploits the egg-laying behavior of mosquitoes to have them disseminate tiny particles of a potent larvicide, pyriproxyfen (PPF), from resting to breeding sites, thus improving coverage. This approach has yielded promising results at small spatial scales, but its wider applicability remains unclear. We conducted a four-month trial within a 20-month study to investigate mosquito-driven dissemination of PPF dust-particles from 100 'dissemination stations' (DSs) deployed in a 7-ha sub-area to surveillance dwellings and sentinel breeding sites (SBSs) distributed over an urban neighborhood of about 50 ha. We assessed the impact of the trial by measuring juvenile mosquito mortality and adult mosquito emergence in each SBS-month. Using data from 1,075 dwelling-months, 2,988 SBS-months, and 29,922 individual mosquitoes, we show that mosquito-disseminated PPF yielded high coverage of dwellings (up to 100%) and SBSs (up to 94.3%). Juvenile mosquito mortality in SBSs (about 4% at baseline) increased by over one order of magnitude during PPF dissemination (about 75%). This led to a >10-fold decrease of adult mosquito emergence from SBSs, from approximately 1,000-3,000 adults/month before to about 100 adults/month during PPF dissemination. By expanding breeding-site coverage and boosting juvenile mosquito mortality, a strategy based on mosquito-disseminated PPF has potential to substantially enhance mosquito control. Sharp declines in adult mosquito emergence can lower vector/host ratios, reducing the risk of disease outbreaks. This approach is a very promising complement to current and novel mosquito control strategies; it will probably be especially relevant for the control of urban disease vectors, such as Aedes and Culex species, that often cause large epidemics.

Aedes aegypti and Ae. albopictus are the vectors of dengue, the most important arboviral disease of humans. To date, Aedes ecology studies have assumed that the vectors are truly absent from sites where they are not detected; since no perfect detection method exists, this assumption is questionable. Imperfect detection may bias estimates of key vector surveillance/control parameters, including site-occupancy (infestation) rates and control intervention effects. We used a modeling approach that explicitly accounts for imperfect detection and a 38-month, 55-site detection/non-detection dataset to quantify the effects of municipality/state control interventions on Aedes site-occupancy dynamics, considering meteorological and dwelling-level covariates. Ae. aegypti site-occupancy estimates (mean 0.91; range 0.79-0.97) were much higher than reported by routine surveillance based on 'rapid larval surveys' (0.03; 0.02-0.11) and moderately higher than directly ascertained with oviposition traps (0.68; 0.50-0.91). Regular control campaigns based on breeding-site elimination had no measurable effects on the probabilities of dwelling infestation by dengue vectors. Site-occupancy fluctuated seasonally, mainly due to the negative effects of high maximum (Ae. aegypti) and minimum (Ae. albopictus) summer temperatures (June-September). Rainfall and dwelling-level covariates were poor predictors of occupancy. The marked contrast between our estimates of adult vector presence and the results from 'rapid larval surveys' suggests, together with the lack of effect of local control campaigns on infestation, that many Aedes breeding sites were overlooked by vector control agents in our study setting. Better sampling strategies are urgently needed, particularly for the reliable assessment of infestation rates in the context of control program management. The approach we present here, combining oviposition traps and site-occupancy models, could greatly contribute to that crucial aim.

Background The purpose of this study was to establish whether the \"bottle assay\", a tool for monitoring insecticide resistance in mosquitoes, can complement and augment the capabilities of the established WHO assay, particularly in resource-poor, logistically challenging environments. Methods Laboratory reared Aedes aegypti and field collected Anopheles darlingi and Anopheles albimanus were used to assess the suitability of locally sourced solvents and formulated insecticides for use with the bottle assay. Using these adapted protocols, the ability of the bottle assay and the WHO assay to discriminate between deltamethrin-resistant Anopheles albimanus populations was compared. The diagnostic dose of deltamethrin that would identify resistance in currently susceptible populations of An. darlingi and Ae. aegypti was defined. The robustness of the bottle assay during a surveillance exercise in the Amazon was assessed. Results The bottle assay (using technical or formulated material) and the WHO assay were equally able to differentiate deltamethrin-resistant and susceptible An. albimanus populations. A diagnostic dose of 10 μg a.i./bottle was identified as the most sensitive discriminating dose for characterizing resistance in An. darlingi and Ae. aegypti . Treated bottles, prepared using locally sourced solvents and insecticide formulations, can be stored for > 14 days and used three times. Bottles can be stored and transported under local conditions and field-assays can be completed in a single evening. Conclusion The flexible and portable nature of the bottle assay and the ready availability of its components make it a potentially robust and useful tool for monitoring insecticide resistance and efficacy in remote areas that require minimal cost tools.

Background Mosquito-borne pathogens pose major public health challenges worldwide. With vaccines or effective drugs still unavailable for most such pathogens, disease prevention heavily relies on vector control. To date, however, mosquito control has proven difficult, with low breeding-site coverage during control campaigns identified as a major drawback. A novel tactic exploits the egg-laying behavior of mosquitoes to have them disseminate tiny particles of a potent larvicide, pyriproxyfen (PPF), from resting to breeding sites, thus improving coverage. This approach has yielded promising results at small spatial scales, but its wider applicability remains unclear. Methodology/Principal Findings We conducted a four-month trial within a 20-month study to investigate mosquito-driven dissemination of PPF dust-particles from 100 'dissemination stations' (DSs) deployed in a 7-ha sub-area to surveillance dwellings and sentinel breeding sites (SBSs) distributed over an urban neighborhood of about 50 ha. We assessed the impact of the trial by measuring juvenile mosquito mortality and adult mosquito emergence in each SBS-month. Using data from 1,075 dwelling-months, 2,988 SBS-months, and 29,922 individual mosquitoes, we show that mosquito-disseminated PPF yielded high coverage of dwellings (up to 100%) and SBSs (up to 94.3%). Juvenile mosquito mortality in SBSs (about 4% at baseline) increased by over one order of magnitude during PPF dissemination (about 75%). This led to a >10-fold decrease of adult mosquito emergence from SBSs, from approximately 1,000-3,000 adults/month before to about 100 adults/month during PPF dissemination. Conclusions/Significance By expanding breeding-site coverage and boosting juvenile mosquito mortality, a strategy based on mosquito-disseminated PPF has potential to substantially enhance mosquito control. Sharp declines in adult mosquito emergence can lower vector/host ratios, reducing the risk of disease outbreaks. This approach is a very promising complement to current and novel mosquito control strategies; it will probably be especially relevant for the control of urban disease vectors, such as Aedes and Culex species, that often cause large epidemics.

Aedes aegypti and Ae. albopictus are the vectors of dengue, the most important arboviral disease of humans. To date, Aedes ecology studies have assumed that the vectors are truly absent from sites where they are not detected; since no perfect detection method exists, this assumption is questionable. Imperfect detection may bias estimates of key vector surveillance/control parameters, including site-occupancy (infestation) rates and control intervention effects. We used a modeling approach that explicitly accounts for imperfect detection and a 38-month, 55-site detection/non-detection dataset to quantify the effects of municipality/state control interventions on Aedes site-occupancy dynamics, considering meteorological and dwelling-level covariates. Ae. aegypti site-occupancy estimates (mean 0.91; range 0.79-0.97) were much higher than reported by routine surveillance based on 'rapid larval surveys' (0.03; 0.02-0.11) and moderately higher than directly ascertained with oviposition traps (0.68; 0.50-0.91). Regular control campaigns based on breeding-site elimination had no measurable effects on the probabilities of dwelling infestation by dengue vectors. Site-occupancy fluctuated seasonally, mainly due to the negative effects of high maximum (Ae. aegypti) and minimum (Ae. albopictus) summer temperatures (June-September). Rainfall and dwelling-level covariates were poor predictors of occupancy. The marked contrast between our estimates of adult vector presence and the results from 'rapid larval surveys' suggests, together with the lack of effect of local control campaigns on infestation, that many Aedes breeding sites were overlooked by vector control agents in our study setting. Better sampling strategies are urgently needed, particularly for the reliable assessment of infestation rates in the context of control program management. The approach we present here, combining oviposition traps and site-occupancy models, could greatly contribute to that crucial aim.