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Background In the absence of vaccines or drugs, insecticides are the mainstay of Aedes-borne disease control. Their utility is challenged by the slow deployment of resources, poor community compliance and inadequate household coverage. Novel application methods are required. Methodology and principal findings A 10% w/w metofluthrin “emanator” that passively disseminates insecticide from an impregnated net was evaluated in a randomized trial of 200 houses in Mexico. The devices were introduced at a rate of 1 per room and replaced at 3-week intervals. During each of 7 consecutive deployment cycles, indoor resting mosquitoes were sampled using aspirator collections. Assessments of mosquito landing behaviours were made in a subset of houses. Pre-treatment, there were no differences in Aedes aegypti indices between houses recruited to the control and treatment arms. Immediately after metofluthrin deployment, the entomological indices between the trial arms diverged. Averaged across the trial, there were significant reductions in Abundance Rate Ratios for total Ae. aegypti, female abundance and females that contained blood meals (2.5, 2.4 and 2.3-times fewer mosquitoes respectively; P, Author summary Insecticidal control tools are heavily relied on for the control of mosquito-borne viruses such as dengue, chikungunya and Zika, but the logistics associated with conventional insecticide use (e.g. space sprays and residual formulations) are challenging. Considerable time and resources are required to treat household interiors; an impediment exacerbated by the difficulty in gaining entrance to households, and sometimes by limited compliance in the community. Another constraint to effective insecticide use is that many mosquito populations are resistant to the chemicals used. Volatile pyrethroids, exhibiting both lethal and behavioural effects on mosquitoes are available in formulations that release insecticides passively to the air, at room temperature. These may be suitable for deployment in houses with the aim of creating “bite-free” spaces. By removing the need for conventional application methods, these devices might be rapidly deployed with minimum disruption to households. This is the first large-scale, randomized control trial to evaluate the entomological impacts of volatile pyrethroids in an urban environment. Using metofluthrin as an example, we confirm that some formulations have a significant impact on Aedes aegypti densities and landing behaviour indoors. These effects occur despite the presence of pyrethroid-resistance alleles associated with conventional insecticide resistance.

Arbovirus infection in Aedes aegypti has historically been quantified from a sample of the adult population by pooling collected mosquitoes to increase detectability. However, there is a significant knowledge gap about the magnitude of natural arbovirus infection within areas of active transmission, as well as the sensitivity of detection of such an approach. We used indoor Ae. aegypti sequential sampling with Prokopack aspirators to collect all mosquitoes inside 200 houses with suspected active ABV transmission from the city of Mérida, Mexico, and tested all collected specimens by RT-PCR to quantify: a) the absolute arbovirus infection rate in individually tested Ae. aegypti females; b) the sensitivity of using Prokopack aspirators in detecting ABV-infected mosquitoes; and c) the sensitivity of entomological inoculation rate (EIR) and vectorial capacity (VC), two measures ABV transmission potential, to different estimates of indoor Ae. aegypti abundance. The total number of Ae. aegypti (total catch, the sum of all Ae. aegypti across all collection intervals) as well as the number on the first 10-min of collection (sample, equivalent to a routine adult aspiration session) were calculated. We individually tested by RT-PCR 2,161 Aedes aegypti females and found that 7.7% of them were positive to any ABV. Most infections were CHIKV (77.7%), followed by DENV (11.4%) and ZIKV (9.0%). The distribution of infected Aedes aegypti was overdispersed; 33% houses contributed 81% of the infected mosquitoes. A significant association between ABV infection and Ae. aegypti total catch indoors was found (binomial GLMM, Odds Ratio > 1). A 10-min indoor Prokopack collection led to a low sensitivity of detecting ABV infection (16.3% for detecting infected mosquitoes and 23.4% for detecting infected houses). When averaged across all infested houses, mean EIR ranged between 0.04 and 0.06 infective bites per person per day, and mean VC was 0.6 infectious vectors generated from a population feeding on a single infected host per house/day. Both measures were significantly and positively associated with Ae. aegypti total catch indoors. Our findings provide evidence that the accurate estimation and quantification of arbovirus infection rate and transmission risk is a function of the sampling effort, the local abundance of Aedes aegypti and the intensity of arbovirus circulation., Author summary Aedes-borne diseases comprise a serious public health burden in many parts of the world, usually affecting low income areas. The ability to detect virus circulation within a population may be key in responding to the threat of outbreaks, providing a cost-effective approach for triggering vector control. Unfortunately, gaps in the knowledge of natural Aedes-borne virus (ABV) infection in Aedes aegypti have led to uncertainties in the consideration of arbovirus surveillance in mosquitoes. Here, we show that the natural infection rate in a mosquito population may not be a function of where Aedes aegypti are, but rather where key human-mosquito contacts occur. Sampling 200 houses with suspected ABV active transmission led us to quantify high virus infection rates in all Aedes aegypti present in the house and use such information to estimate the sensitivity of indoor aspiration with Prokopack devices and two measures of ABV transmission potential. Our findings provide evidence that the accurate quantification of arbovirus infection rate and transmission risk is a function of the sampling effort, the local abundance of Aedes aegypti and the intensity of arbovirus circulation. Results from this study are relevant to understand the value of virus testing of vector populations, and for the design of entomological endpoints relevant for epidemiological trials quantifying the impact of vector control on ABVs.

Challenges in maintaining high effectiveness of classic vector control in urban areas has renewed the interest in indoor residual spraying (IRS) as a promising approach for Aedes-borne disease prevention. While IRS has many benefits, application time and intrusive indoor applications make its scalability in urban areas difficult. Modifying IRS to account for Ae. aegypti resting behavior, named targeted IRS (TIRS, spraying walls below 1.5 m and under furniture) can reduce application time; however, an untested assumption is that modifications to IRS will not negatively impact entomological efficacy. We conducted a comparative experimental study evaluating the residual efficacy of classically-applied IRS (as developed for malaria control) compared to two TIRS application methods using a carbamate insecticide against a pyrethroid-resistant, field-derived Ae. aegypti strain. We performed our study within a novel experimental house setting (n = 9 houses) located in Merida (Mexico), with similar layouts and standardized contents. Classic IRS application (insecticide applied to full walls and under furniture) was compared to: a) TIRS: insecticide applied to walls below 1.5 m and under furniture, and b) Resting Site TIRS (RS-TIRS): insecticide applied only under furniture. Mosquito mortality was measured eight times post-application (out to six months post-application) by releasing 100 Ae. aegypti females /house and collecting live and dead individuals after 24 hrs exposure. Compared to Classic IRS, TIRS and RS-TIRS took less time to apply (31% and 82% reduction, respectively) and used less insecticide (38% and 85% reduction, respectively). Mortality of pyrethroid-resistant Ae. aegypti did not significantly differ among the three IRS application methods up to two months post application, and did not significantly differ between Classic IRS and TIRS up to four months post application. These data illustrate that optimizing IRS to more efficiently target Ae. aegypti can both reduce application time and insecticide volume with no apparent reduction in entomological efficacy., Author summary Vector control is the primary strategy for managing Aedes aegypti and reducing transmission of Aedes-borne diseases; however, the indoor resting behavior of Ae. aegypti and the evolution of insecticide resistance reduces the effectiveness of many vector control tactics. Indoor residual spraying (IRS) is effective against Ae. aegypti, but lengthy application time makes IRS difficult to scale within urban environments. We compared the application and entomological efficacy of Classic IRS against two novel Aedes-targeting IRS application methods (Targeted IRS [TIRS]- insecticide applied to walls below 1.5 m and under furniture and Resting Site TIRS [RS-TIRS]- insecticide applied only under furniture) within experimental houses using a carbamate insecticide. Both TIRS and RS-TIRS took less time to apply and used less insecticide compared to Classic IRS. Mortality of pyrethroid-resistant Ae. aegypti did not differ among treatments out to two months post-application, and there was no difference in mortality between Classic IRS and TIRS out to four months post-application. These data provide evidence that IRS application methods can be improved to take less time and insecticide yet not lose entomological efficacy, making TIRS more scalable within urban environments. However, larger field studies with epidemiologic endpoints are needed to further assess the efficacy of these modified TIRS techniques.