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Background There is an increased need to mitigate the emergence of insecticide resistance and incorporate new formulations and modes of application to control the urban vector Aedes aegypti. Most research and development of insecticide formulations for the control of Ae. aegypti has focused on their peridomestic use as truck-mounted ULV-sprays or thermal fogs despite the widespread knowledge that most resting Ae. aegypti are found indoors. A recent modification of indoor residual spraying (IRS), termed targeted IRS (TIRS) works by restricting applications to 1.5 m down to the floor and on key Ae. aegypti resting sites (under furniture). TIRS also opens the possibility of evaluating novel residual insecticide formulations currently being developed for malaria IRS. Methods We evaluated the residual efficacy of chlorfenapyr, formulated as Sylando 240SC, for 12 months on free-flying field-derived pyrethroid-resistant Ae. aegypti using a novel experimental house design in Merida, Mexico. On a monthly basis, 600 female Ae. aegypti were released into the houses and left indoors with access to sugar solution for 24 hours. After the exposure period, dead and alive mosquitoes were counted in houses treated with chlorfenapyr as well as untreated control houses to calculate 24-h mortality. An evaluation for these exposed cohorts of surviving mosquitoes was extended up to seven days under laboratory conditions to quantify “delayed mortality”. Results Mean acute (24-h) mortality of pyrethroid-resistant Ae. aegypti ranged 80–97% over 5 months, dropping below 30% after 7 months post-TIRS. If delayed mortality was considered (quantifying mosquito mortality up to 7 days after exposure), residual efficacy was above 90% for up to 7 months post-TIRS application. Generalized Additive Mixed Models quantified a residual efficacy of chlorfenapyr of 225 days (ca. 7.5 months). Conclusions Chlorfenapyr represents a new option for TIRS control of Ae. aegypti in urban areas, providing a highly-effective time of protection against indoor Ae. aegypti females of up to 7 months., Author summary Vector control (VC) for managing Aedes aegypti and reducing transmission of Aedes-borne diseases is largely focused on peridomestic insecticide applications. However, the indoor resting behavior of Ae. aegypti and the acceleration of insecticide resistance owed to reduced modes of action have diminished the effectiveness of many VC tools. A targeted Indoor residual spraying (TIRS) modality in experimental housing units was employed to investigate the potential of chlorfenapyr, a pyrrole-class insecticide with known effectiveness to resistant mosquito species. This was the first investigation for chlorfenapyr use against locally resistant Ae. aegypti (Merida, Mexico) with this approach. Two treatment arms were investigated in the present study: TIRS and a control house where only water was sprayed. A comparison of entomological efficacy for TIRS applied to interior perimeter walls below 1.5 m with chlorfenapyr (formulated as Sylando 240SC) at 250 mg/m2 over 12 months was assessed. TIRS chlorfenapyr treatments were highly efficacious and led to acute mortalities (after 24 exposure) above 80% up to 5 months; delayed mortalities (to Ae. aegypti) were monitored over seven days post exposures vs untreated controls. When delayed mortality was considered, residual efficacy of chlorfenapyr extended to 7 months. These data provide evidence that TIRS chlorfenapyr is an effective Aedes management tool that surpassed efficacy profiles for other TIRS insecticides that have been previously reported with this method. Further, Chlorfenapyr emerges as a novel addition to Ae. aegypti VC, and future studies should focus on its effectiveness and residual power as part of Phase II-III TIRS trials.

After the tropical storm Cristobal, we performed special adult entomological collections in the peri-domicile of 35 houses from 25 neighborhoods of Mérida, Yucatan, Mexico in response to complaints from the community about an increased nuisance due to an abundance of mosquitoes. A total of 1,275 specimens from four genera and 13 species were collected: Aedes taeniorhynchus (92%), Culex quinquefasciatus (72%), Aedes aegypti (72%), Psorophora mexicana (36%), Psorophora cyanescens (32%), Aedes scapularis (24%), Culex nigripalpus (24%), Aedes albopictus (8%), Psorophora ferox (4%), Haemagogus equinus (4%), Aedes trivittatus (4%), Culex coronator (4%), Culex iolambdis (4%). From these collections, the increased mosquito nuisance was mainly the result of invasive species such as Aedes taeniorhynchus and Psorophora. City wide, vehicle mounted ULV spraying was performed by the MoH and the municipality of Merida to control adult mosquito populations. We report Culex iolambdis for the first time in Merida and Psorophora mexicana for the state of Yucatan.

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.

Background The integration of house-screening and long-lasting insecticidal nets, known as insecticide-treated screening (ITS), can provide simple, safe, and low-tech Aedes aegypti control. Cluster randomised controlled trials in two endemic localities for Ae. aegypti of south Mexico, showed that ITS conferred both, immediate and sustained (~2 yr) impact on indoor-female Ae. aegypti infestations. Such encouraging results require further validation with studies quantifying more epidemiologically-related endpoints, including arbovirus infection in Ae. aegypti. We evaluated the efficacy of protecting houses with ITS on Ae. aegypti infestation and arbovirus infection during a Zika outbreak in Merida, Yucatan, Mexico. Methodology/Principal findings A two-arm cluster-randomised controlled trial evaluated the entomological efficacy of ITS compared to the absence of ITS (with both arms able to receive routine arbovirus vector control) in the neighbourhood Juan Pablo II of Merida. Cross-sectional entomological surveys quantified indoor adult mosquito infestation and arbovirus infection at baseline (pre-ITS installation) and throughout two post-intervention (PI) surveys spaced at 6-month intervals corresponding to dry/rainy seasons over one year (2016–2017). Household-surveys assessed the social reception of the intervention. Houses with ITS were 79–85% less infested with Aedes females than control houses up to one-year PI. A similar significant trend was observed for blood-fed Ae. aegypti females (76–82%). Houses with ITS had significantly less infected female Ae. aegypti than controls during the peak of the epidemic (OR = 0.15, 95%CI: 0.08–0.29), an effect that was significant up to a year PI (OR = 0.24, 0.15–0.39). Communities strongly accepted the intervention, due to its perceived mode of action, the prevalent risk for Aedes-borne diseases in the area, and the positive feedback from neighbours receiving ITS. Conclusions/Significance We show evidence of the protective efficacy of ITS against an arboviral disease of major relevance, and discuss the relevance of our findings for intervention adoption., Author summary We evaluated the efficacy of protecting houses with insecticide-treated nets permanently fixed with aluminium frames on external doors and windows on Ae. aegypti infestation and arbovirus infection during a Zika outbreak in Merida, Yucatan, Mexico. Houses protected with screens were ≈80% less infested with Aedes females and very importantly, had significantly less infected female Ae. aegypti during the peak of the epidemic. Communities strongly accepted the intervention, due to its perceived mode of action, the prevalent risk for Aedes-borne diseases in the area, and the positive feedback from neighbours. House screening provides a simple, affordable sustainable method to reduce human-vector contact inside houses and can protect against dengue, chikungunya and Zika.

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.

Background There is a need for effective methods to control Aedes aegypti and prevent the transmission of dengue, chikungunya, yellow fever and Zika viruses. Insecticide treated screening (ITS) is a promising approach, particularly as it targets adult mosquitoes to reduce human-mosquito contact. Methodology/Principal findings A cluster-randomised controlled trial evaluated the entomological efficacy of ITS based intervention, which consisted of the installation of pyrethroid-impregnated long-lasting insecticide-treated netting material fixed as framed screens on external doors and windows. A total of 10 treatment and 10 control clusters (100 houses/cluster) were distributed throughout the city of Merida, Mexico. Cross-sectional entomological surveys quantified indoor adult mosquito infestation at baseline (pre-intervention) and throughout four post-intervention (PI) surveys spaced at 6-month intervals corresponding to dry/rainy seasons over two years (2012–2014). A total of 844 households from intervention clusters (86% coverage) were protected with ITS at the start of the trial. Significant reductions in the indoor presence and abundance of Ae. aegypti adults (OR = 0.48 and IRR = 0.45, P, Author summary An ITS based intervention, screened houses with insecticide-treated netting (long-lasting insecticidal nets permanently fixed with aluminium frames on external doors and windows), in Merida city, Mexico. Screened houses had >50% less chance of having Ae. aegypti mosquitoes indoors and importantly, >50% fewer female Ae. aegypti in comparison with non-screened houses (control houses). The impact was sustained over 2 years. House screening could provide a feasible sustainable method to reduce human-vector contact inside houses and could potentially be scaled up to protect against mosquito-borne diseases, particularly in locations where the simultaneous transmission, or threat, of dengue, chikungunya, yellow fever and Zika is occurring.

The operational impact of deltamethrin resistance on the efficacy of indoor insecticide applications to control Aedes aegypti was evaluated in Merida, Mexico. A randomized controlled trial quantified the efficacy of indoor residual spraying (IRS) against adult Ae. aegypti in houses treated with either deltamethrin (to which local Ae. aegypti expressed a high degree of resistance) or bendiocarb (to which local Ae. aegypti were fully susceptible) as compared to untreated control houses. All adult Ae. aegypti infestation indices during 3 months post-spraying were significantly lower in houses treated with bendiocarb compared to untreated houses (odds ratio, Author summary Insecticide resistance has emerged as a worrisome outlook for the implementation of insecticide-only approaches. Particularly for Aedes aegypti, it is generally argued that this rapid rise of insecticide resistance may compromise the effectiveness of control programs. Yet, empirical evidence of such negative operational impact is lacking. This article provides quantitative evidence of the impact of insecticide resistance, primarily to the pyrethroid insecticide deltamethrin, on the entomological effectiveness of interventions against Aedes aegypti mosquitoes. Within an area of high resistance to pyrethroids, performing high-quality insecticide applications using deltamethrin had no entomological impact in comparison to the application of an insecticide to which Ae. aegypti was susceptible. Deltamethrin did not protect against resistant Ae. aegypti, whereas the application of bendiocarb led to an average efficacy of 60% during a 3-month period. This study provides strong evidence of the dramatic operational treatment failure that can occur when the background insecticide resistance level of Ae. aegypti populations is not taken into consideration when performing vector control.