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Insecticide-based approaches remain a key pillar for Aedes-borne virus (ABV, dengue, chikungunya, Zika) control, yet they are challenged by the limited effect of traditional outdoor insecticide campaigns responding to reported arboviral cases and by the emergence of insecticide resistance in mosquitoes. A three-arm Phase II unblinded entomological cluster randomized trial was conducted in Merida, Yucatan State, Mexico, to quantify the entomological impact of targeted indoor residual spraying (TIRS, application of residual insecticides in Ae. aegypti indoor resting sites) applied preventively 2 months before the beginning of the arbovirus transmission season. Trial arms involved the use of two insecticides with unrelated modes of action (Actellic 300CS, pirimiphos-methyl, and SumiShield 50WG, clothianidin) and a control arm where TIRS was not applied. Entomological impact was quantified by Prokopack adult collections performed indoors during 10 min per house. Regardless of the insecticide, conducting a preventive TIRS application led to significant reductions in indoor Ae. aegypti densities, which were maintained at the same levels as in the low arbovirus transmission period (Actellic 300CS reduced Ae. aegypti density up to 8 months, whereas SumiShield 50WG up to 6 months). The proportional reduction in Ae. aegypti abundance in treatment houses compared to control houses was 50–70% for Actellic 300CS and 43–63% for SumiShield 50WG. Total operational costs including insecticide ranged from US$4.2 to US$10.5 per house, depending on the insecticide cost. Conducting preventive residual insecticide applications can maintain Ae. aegypti densities at low levels year-round with important implications for preventing ABVs in the Americas and beyond.

Objective To evaluate the protective effect of house-screening (HS) on indoor Ae. aegypti infestation, abundance and arboviral infection in Merida, Mexico. Methods In 2019, we performed a cluster randomized controlled trial (6 control and 6 intervention areas: 100 households/area). Intervention clusters received permanently fixed fiberglass HS on all windows and doors. The study included two cross-sectional entomologic surveys, one baseline (dry season in May 2019) and one post-intervention (PI, rainy season between September-October 2019). The presence and number of indoor Aedes females and blood-fed females (indoor mosquito infestation) as well as arboviral infections with dengue (DENV) and Zika (ZIKV) viruses were evaluated in a subsample of 30 houses within each cluster. Results HS houses had significatively lower risk for having Aedes aegypti female mosquitoes (odds ratio [OR]=0.56, 95% C. I. 0.33-0.97, P=0.04) and blood-fed females (OR=0.53, 95% C. I. 0.28-0.97, P=0.04) than unscreened households from the control arm. Compared to control houses, HS houses had significantly lower indoor Ae. aegypti abundance (rate ratio [RR]=0.50, 95% C. I. 0.30-0.83, P=0.01), blood-fed Ae. aegypti females (RR=0.48, 95% C. I. 0.27-0.85, P=0.01) and female Ae. aegypti positive for arboviruses (OR=0.29, 95% CI 0.10-0.86, P=0.02). The estimated intervention efficacy in reducing Ae. aegypti arbovirus infection was 71%. Conclusions These results provide evidence supporting the use of HS as an effective pesticide-free method to control house infestations with Aedes aegypti and reduce the transmission of Aedes-transmitted viruses such as DENV, chikungunya (CHIKV), and ZIKV.

We conducted a baseline characterization of the abundance and seasonality of Aedes aegypti (Linnaeus, 1762)—a vector of dengue, chikungunya, and Zika—in two suburban localities of Yucatan, Mexico, as the first step in the implementation of an integrated vector management (IVM) plan combining ‘traditional Aedes control’ (source reduction/truck-mounted ultra-low volume [ULV] spraying) and incompatible insect technique/sterile insect technique for population suppression in Yucatan, Mexico. Weekly entomological collections with ovitraps and BG-sentinel traps were performed in 1-ha quadrants of both localities for 1 yr. Three distinct periods/phases were identified, closely associated with precipitation: 1) a phase of low population abundance during the dry season (weekly average of Aedes eggs per ovitrap and adults per BG trap = 15.51 ± 0.71 and 10.07 ± 0.88, respectively); 2) a phase of population growth and greatest abundance of Aedes (49.03 ± 1.48 eggs and 25.69 ± 1.31 adults) during the rainy season; and finally 3) a phase of decline among populations (20.91 ± 0.97 eggs and 3.24 ± 0.21 adults) after the peak of the rainy season. Seasonal abundance and dynamics of Ae. aegypti populations suggest that it is feasible to develop and implement time-specific actions as part of an IVM approach incorporating integrating novel technologies (such as rear-and-release of Wolbachia-infected males) with classic (insecticide-based) approaches implemented routinely for vector control. In agreement with the local vector control program, we propose a pilot IVM strategy structured in a preparation phase, an attack phase with traditional vector control, and a suppression phase with inundative releases, which are described in this paper.

We report results of susceptibility tests for Aedes taeniorhynchus from 2 localities of Yucatan State, Mexico, to different insecticides. The Centers for Disease Control and Prevention bottle bioassays were performed using the active ingredients of 3 pyrethroids, 2 organophosphates, and 1 carbamate: permethrin (15 μg/ml), deltamethrin (10 μg/ml), alpha-cypermethrin (10 μg/ml), malathion (50 μg/ml), chlorpyrifos (85 μg/ml), and bendiocarb (12.5 μg/ml). The mortality recorded at the diagnostic time of exposure (30 min) was 100% with all insecticides evaluated and for both populations. These results suggest complete susceptibility to the 3 chemical groups generally used for urban Ae. aegypti mosquito control.

Aedes-borne viruses (ABVs) such as dengue (DENV), chikungunya (CHIKV), and Zika (ZIKV) contribute significantly to the global burden of infectious diseases, disproportionately affecting disadvantaged populations from tropical and subtropical urban areas. ABVs can be transmitted from female mosquitoes to their progeny by vertical transmission via transovarial and/or trans-egg vertical transmission and contribute to the maintenance of infected-mosquito populations year-round in endemic regions. This study describes the natural infection rate of DENV, CHIKV, and ZIKV in field-caught male Aedes (Sergentomyia) aegypti (Linnaeus) mosquitoes from Mérida, Yucatán, México, as a proxy for the occurrence of vertical virus transmission. We used indoor sequential sampling with Prokopack aspirators to collect all mosquitoes inside houses from ABV hotspots areas. Collections were performed in a DENV and CHIKV post-epidemic phase and during a period of active ZIKV transmission. We individually RT-qPCR tested all indoor collected Ae. aegypti males (1,278) followed by Sanger sequencing analysis for final confirmation. A total of 6.7% male mosquitoes were positive for ABV (CHIKV = 5.7%; DENV = 0.9%; ZIKV = 0.1%) and came from 21.0% (30/143) houses infested with males. Most ABV-positive male mosquitoes were positive for CHIKV (84.8%). The distribution of ABV-positive Ae. aegypti males was aggregated in a few households, with two houses having 11 ABV-positive males each. We found a positive association between ABV-positive males and females per house. These findings suggested the occurrence of vertical arbovirus transmission within the mosquito populations in an ABV-endemic area and, a mechanism contributing to viral maintenance and virus re-emergence among humans in post-epidemic periods.

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.

In recent years, Aedes albopictus has become the most important invasive mosquito species worldwide. In 2018, Ae. albopictus was found in a suburban area of Merida, one of the cities with the highest number of arbovirus cases in Mexico in the last 10 years. As Ae. albopictus continues its range expansion, there is a need to monitor its susceptibility to existing insecticide classes, since countries like Mexico currently do not consider Ae. albopictus in its insecticide management programs. In order to determine its susceptibility to the insecticides usually applied by the vector control program in Mexico, the Centers for Disease Control and Prevention bottle bioassays were performed on individuals from established population of Ae. albopictus from Merida, Yucatan, Mexico. Results suggested that the population recently found in the suburban area of Merida is susceptible to permethrin, deltamethrin, chlorpyrifos, malathion, bendiocarb, and propoxur. Further studies of insecticide resistance using biochemical and molecular tools together with more knowledge of the biology and ecology of this species are necessary to generate specific and efficient control strategies in Mexico.

A cluster-randomized controlled trial quantified the entomological efficacy of aerial ultra-low volume (AULV) applications of the insecticide chlorpyrifos against Aedes aegypti in Puerto Vallarta, México, during November–October 2017. The trial involved 16 large (1 × 1 km) clusters distributed between treatment-control arms. Primary endpoint was the abundance of Ae. aegypti indoors (total adults, females, and blood-fed females) collected using Prokopack aspirators. After four consecutive weekly cycles of AULV, all adult Ae. aegypti infestation indices were significantly lower in the treatment arm (OR and IRR ≤ 0.28). Efficacy in reducing indoor Ae. aegypti increased with each weekly application cycle from 30 to 73% (total adults), 33 to 76% (females), and 45.5 to 89% (blood-fed females). Entomological indices remained significantly lower in the treatment arm up to 2 wk after the fourth spraying round. Performing AULV spraying can have significant and lasting entomological impact on Ae. aegypti as long as multiple (ideally four) spray cycles are implemented using an effective insecticide.

Summary Background Effective Aedes aegypti control is limited, in part, by the difficulty in achieving sufficient intervention coverage. To maximise the effect of vector control, areas with persistently high numbers of Aedes-borne disease cases could be identified and prioritised for preventive interventions. We aimed to identify persistent Aedes-borne disease hotspots in cities across southern Mexico. Methods In this spatial analysis, geocoded cases of dengue, chikungunya, and Zika from nine endemic Mexican cities were aggregated at the census-tract level. We included cities that were located in southern Mexico (the arbovirus endemic region of Mexico), with a high burden of dengue cases (ie, more than 5000 cases reported during a 10-year period), and listed as high priority for the Mexican dengue control and prevention programme. The Getis-Ord Gi*(d) statistic was applied to yearly slices of the dataset to identify spatial hotspots of each disease in each city. We used Kendall's W coefficient to quantify the agreement in the distribution of each virus. Findings 128 507 dengue, 4752 chikungunya and 25 755 Zika clinical cases were reported between Jan 1, 2008, and Dec 31, 2016. All cities showed evidence of transmission heterogeneity, with a mean of 17·6% (SD 4·7) of their total area identified as persistent disease hotspots. Hotspots accounted for 25·6% (SD 9·7; range 12·8–43·0) of the population and 32·1% (10·5; 19·6–50·5) of all Aedes-borne disease cases reported. We found an overlap between hotspots of 61·7% for dengue and Zika and 53·3% for dengue and chikungunya. Dengue hotspots in 2008–16 were significantly associated with dengue hotspots detected during 2017–20 in five of the nine cities. Heads of vector control confirmed hotspot areas as problem zones for arbovirus transmission. Interpretation This study provides evidence of the overlap of Aedes-borne diseases within geographical hotspots and a methodological framework for the stratification of arbovirus transmission risk within urban areas, which can guide the implementation of surveillance and vector control. Funding USAID, the US Centers for Disease Control and Prevention, the Canadian Institutes of Health Research, International Development Research Centre, Fondo Mixto CONACyT (Mexico)-Gobierno del Estado de Yucatan, and the US National Institutes of Health. Translation For the Spanish translation of the abstract see Supplementary Materials section.

Background With the recent occurrence of locally transmitted Aedes-borne viruses in the continental United States and Europe, and a lack of effective vaccines, new approaches to control Aedes aegypti and Aedes albopictus are needed. In sub-tropical urban settings in the US, Ae. albopictus is a dominant nuisance and arbovirus vector species. Unfortunately, the vector control toolbox against Ae. albopictus is not as well developed as for Ae. aegypti. Here, we evaluate the efficacy, longevity, and range of protectiveness of a novel passive metofluthrin emanator (10% active ingredient in a polyethylene mesh) against Ae. albopictus indoors and outdoors. Methods Four studies were conducted comparing the presence of the metofluthrin emanator to a control lacking emanator with interest in quantifying efficacy by human landing counts. Studies evaluated the effect of an emanator at varying distances from one or more human volunteers indoors and outdoors. Efficacy of emanators over time since activation was also evaluated. Results Mixed-effects models determined that sitting in close proximity to an emanator reduced landings by 89.5% outdoors and by 74.6% indoors. The emanator was determined protective when located immediately next to a human volunteer outdoors but not uniformly protective when located further away. The emanator was protective at all tested distances from the device indoors. Mortality of mosquitoes exposed to metofluthrin emanators was ~2x higher than those who were not exposed in indoor conditions. Finally, a Generalized Additive Model determined that emanators used continuously outdoors lost their effect after 2.5 weeks and stopped inducing paralysis in mosquitoes after 3.8 weeks of use. Conclusions We show strong and lasting efficacy of 10% metofluthrin emanators against field Ae. albopictus both in indoor and outdoor conditions. Metofluthrin emanators can protect people from Ae. albopictus bites, representing a viable option for reducing human-mosquito contacts at home and beyond.

Aedes aegypti and Aedes albopictus vector dengue, chikungunya and Zika viruses. With both species expanding their global distributions at alarming rates, developing effective surveillance equipment is a continuing priority for public health researchers. Sound traps have been shown, in limited testing, to be highly species-specific when emitting a frequency corresponding to a female mosquito wingbeat. Determining male mosquito capture rates in sound traps based on lure frequencies in endemic settings is the next step for informed deployment of these surveillance tools. We field-evaluated Male Aedes Sound Traps (MASTs) set to either 450 Hz, 500 Hz, 550 Hz or 600 Hz for sampling Aedes aegypti and/or Aedes albopictus and compared catch rates to BG-Sentinel traps within Pacific (Madang, Papua New Guinea) and Latin American (Molas, Mexico and Orange Walk Town, Belize) locations. MASTs set to 450–550 Hz consistently caught male Ae. aegypti at rates comparable to BG-Sentinel traps in all locations. A peak in male Ae. albopictus captures in MASTs set at 550 Hz was observed, with the lowest mean abundance recorded in MASTs set to 450 Hz. While significantly higher abundances of male Culex were sampled in MASTs emitting lower relative frequencies in Molas, overall male Culex were captured in significantly lower abundances in the MASTs, relative to BG-Sentinel traps within all locations. Finally, significant differences in rates at which male Aedes and Culex were positively detected in trap-types per weekly collections were broadly consistent with trends in abundance data per trap-type. MASTs at 550 Hz effectively captured both male Ae. aegypti and Ae. albopictus while greatly reducing bycatch, especially male Culex, in locations where dengue transmission has occurred. This high species-specificity of the MAST not only reduces staff-time required to sort samples, but can also be exploited to develop an accurate smart-trap system—both outcomes potentially reducing public health program expenses., Author summary The continued global expansion of Aedes aegypti and Aedes albopictus, which transmit viruses causing dengue and Zika, necessitates the further development of effective mosquito traps for monitoring and predicting transmission risk. Mosquito sound traps attract male mosquitoes by producing a frequency which mimics the wingbeat sound of female mosquitoes. As few insects will respond to such sounds, these traps can be highly species-specific, saving significant person-time to sort samples, potentially paving the way for more accurate smart-trap systems and reduced surveillance costs. To date, limited research has occurred within field-settings investigating the capture rates of male mosquitoes in sound traps set to different frequencies. To this point, we performed trials in dengue endemic environments to quantify capture rates of mosquitoes in Male Aedes Sound Traps set to different frequencies in Papua New Guinea, Mexico and Belize. We found that both male Ae. aegypti and Ae. albopictus responded positively to frequencies between 450 and 600 Hz. Additionally, male Culex mosquitoes were also caught, but at lower abundances in traps set to higher sound frequencies. These traps, when set to 550 Hz, effectively capture male Ae. aegypti or Ae. albopictus with limited bycatch.

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.

Background Current urban vector control strategies have failed to contain dengue epidemics and to prevent the global expansion of Aedes-borne viruses (ABVs: dengue, chikungunya, Zika). Part of the challenge in sustaining effective ABV control emerges from the paucity of evidence regarding the epidemiological impact of any Aedes control method. A strategy for which there is limited epidemiological evidence is targeted indoor residual spraying (TIRS). TIRS is a modification of classic malaria indoor residual spraying that accounts for Aedes aegypti resting behavior by applying residual insecticides on exposed lower sections of walls ( Methods/design We are pursuing a two-arm, parallel, unblinded, cluster randomized controlled trial to quantify the overall efficacy of TIRS in reducing the burden of laboratory-confirmed ABV clinical disease (primary endpoint). The trial will be conducted in the city of Merida, Yucatan State, Mexico (population ~ 1million), where we will prospectively follow 4600 children aged 2–15 years at enrollment, distributed in 50 clusters of 5 × 5 city blocks each. Clusters will be randomly allocated (n = 25 per arm) using covariate-constrained randomization. A “fried egg” design will be followed, in which all blocks of the 5 × 5 cluster receive the intervention, but all sampling to evaluate the epidemiological and entomological endpoints will occur in the “yolk,” the center 3 × 3 city blocks of each cluster. TIRS will be implemented as a preventive application (~ 1–2 months prior to the beginning of the ABV season). Active monitoring for symptomatic ABV illness will occur through weekly household visits and enhanced surveillance. Annual sero-surveys will be performed after each transmission season and entomological evaluations of Ae. aegypti indoor abundance and ABV infection rates monthly during the period of active surveillance. Epidemiological and entomological evaluation will continue for up to three transmission seasons. Discussion The findings from this study will provide robust epidemiological evidence of the efficacy of TIRS in reducing ABV illness and infection. If efficacious, TIRS could drive a paradigm shift in Aedes control by considering Ae. aegypti behavior to guide residual insecticide applications and changing deployment to preemptive control (rather than in response to symptomatic cases), two major enhancements to existing practice. Trial registration ClinicalTrials.gov NCT04343521. Registered on 13 April 2020. The protocol also complies with the WHO International Clinical Trials Registry Platform (ICTRP) (Additional file 1). Primary sponsor National Institutes of Health, National Institute of Allergy and Infectious Diseases (NIH/NIAID).

This study reports the results of a molecular screening for Wolbachia (Wb) infection in Aedes albopictus (Skuse) populations recently established in the Yucatan Peninsula, Mexico. To do so, collections of free-flying adults with BG traps and emerged adults from eggs after ovitrap field collections were performed in three suburban localities of the city of Merida, Yucatan. Overall, local populations of Ae. albopictus present a natural Wb infection rate of ~40% (18 of 45). Wb infection was detected in both field-collected adults (76.5%, 13 of 17) and eggs reared (17.8%, 5 of 28) and in 37.9% (11/29) of females and 43.7% (7/16) of male Ae. albopictus mosquitoes. An initial screening for Wolbachia strain typing showed that native Ae. albopictus were naturally coinfected with both wAlbA and wAlbB strains. The knowledge of the prevalence and diversity of Wolbachia strains in local populations of Aedes mosquitoes is part of the baseline information required for current and future Wolbachia-based vector control approaches to be conducted in Mexico.

Vector control methods that mobilize and impact rapidly during dengue, Zika, and chikungunya outbreaks are urgently needed in urban contexts. We investigated whether one person using a handheld aerosolized insecticide could achieve efficacy levels comparable to targeted indoor residual spraying (TIRS), using pyrethroid-resistant Aedes aegypti in a semi-field setting with experimental houses in Mexico. The insecticide product (H24, a carbamate and pyrethroid mixture), available over-the-counter locally, was sprayed only on known Ae. aegypti–resting surfaces, for example, walls less than 1.5 m and dark hidden areas. In six identical houses with paired bedrooms, one bedroom was treated, and the other remained an untreated control. Each week for 8 weeks, 100 female pyrethroid-resistant Ae. aegypti were released in each bedroom and followed up daily. Mortality rates in treated bedrooms exceeded 90% for at least 2 weeks, and more than 80% (89.2; 95% CI: 79.98–98.35) for 3 weeks or more. Mortality rates in control houses were zero. Results demonstrate that the immediate impact of TIRS can be delivered by one person using existing products, at an estimated cost for the average household in Mexico of under US$3 per month. Triggered by early outbreak signs, dissemination via community hubs and mass/social media of instructions to treat the home immediately, with monthly re-treatment thereafter, provides a simple means to engage and empower householders. Compatible with integrated vector management strategies, it enables self-protection even if existing agencies falter, a situation exemplified by the potential impact on vector control of the restrictions imposed during the 2020 COVID-19 pandemic.

Introduction: The vector-borne diseases program in México has an established network of ovitraps for entomological surveillance of Aedes spp. In response to reports of Aedes albopictus in the periphery of Mérida, the state capital of Yucatán, the Ministry of Health increased the specificity of this surveillance. Objective: To describe the presence and distribution of Ae. albopictus in Mérida and its relative abundance compared to Aedes aegypti in ovitraps of the vector control program. Materials and methods: During October, 2019, 91 ovitraps were randomly selected from 31 neighborhoods of Mérida. Mosquitoes were reared at the insectary of the Collaborative Unit for Entomological Bioassays of the Autonomous University of Yucatán from eggs collected in the field. Relative abundance was determined for adult individuals of each identified species and neighborhood. Results: 32 % of the neighborhoods were positive for Ae. albopictus and 100 % for Ae. aegypti. A total of 28 adults of Ae. albopictus (10 females and 18 males) were obtained from ovitraps. No correlation was observed between the abundance of Ae. aegypti and Ae. albopictus for both adults and females (p0.05) at the neighborhood level. Conclusions: The results confirm that Ae. albopictus coexisted with Ae. aegypti in Mérida at the time of the study. The low relative abundance suggests that Ae. albopictus was in the initial phase of invasion.Introducción. El programa de enfermedades transmitidas por vectores en México tiene una red establecida de ovitrampas para la vigilancia entomológica de Aedes spp. Los servicios de salud del estado de Yucatán, en respuesta a reportes de Aedes albopictus en la periferia de Mérida, capital del estado, incrementaron la especificidad de dicha vigilancia. Objetivo. Describir la presencia y distribución de Ae. albopictus en Mérida y su abundancia relativa comparada con Aedes aegypti, en ovitrampas del programa de control de vectores. Materiales y métodos. Durante octubre de 2019, se seleccionaron al azar 91 ovitrampas en 31 barrios de Mérida. Los mosquitos adultos se obtuvieron del insectario de la Unidad Colaborativa para Bioensayos Entomológicos de la Universidad Autónoma de Yucatán a partir de huevos recolectados en campo. Se determinó la abundancia relativa de individuos adultos de cada especie identificada y por barrios evaluados. Resultados. En el 32 % de los barrios muestreados, se detectó Ae. albopictus y, en todos ellos, Ae. aegypti. Se recolectaron 28 adultos de Ae. albopictus (10 hembras y 18 machos) en las ovitrampas. No se observó correlación entre la abundancia de adultos ni de hembras Ae. aegypti y Ae. albopictus por barrio (p0,05). Conclusiones. Los resultados confirmaron que Ae. albopictus estaba coexistiendo con Ae. aegypti en Mérida en el momento del estudio. La baja abundancia relativa sugiere que Ae. albopictus se encontraba en la fase inicial de invasión.

BackgroundThe combination ofWolbachia-based incompatible insect technique (IIT) and radiation-based sterile insect technique (SIT) can be used for population suppression ofAedes aegypti. Our main objective was to evaluate whether open-field mass-releases ofwAlbB-infectedAe.aegyptimales, as part of an Integrated Vector Management (IVM) plan led by the Mexican Ministry of Health, could suppress natural populations ofAe.aegyptiin urbanized settings in south Mexico.Methodology/Principal findingsWe implemented a controlled before-and-after quasi-experimental study in two suburban localities of Yucatan (Mexico): San Pedro Chimay (SPC), which received IIT-SIT, and San Antonio Tahdzibichén used as control. Release ofwAlbBAe.aegyptimales at SPC extended for 6 months (July-December 2019), covering the period of higherAe.aegyptiabundance. Entomological indicators included egg hatching rates and outdoor/indoor adult females collected at the release and control sites. Approximately 1,270,000 lab-producedwAlbB-infectedAe.aegyptimales were released in the 50-ha treatment area (2,000wAlbBAe.aegyptimales per hectare twice a week in two different release days, totaling 200,000 male mosquitoes per week). The efficacy of IIT-SIT in suppressing indoor femaleAe.aegyptidensity (quantified from a generalized linear mixed model showing a statistically significant reduction in treatment versus control areas) was 90.9% a month after initiation of the suppression phase, 47.7% two months after (when number of released males was reduced in 50% to match local abundance), 61.4% four months after (when initial number of released males was re-established), 88.4% five months after and 89.4% at six months after the initiation of the suppression phase. A proportional, but lower, reduction in outdoor femaleAe.aegyptiwas also quantified (range, 50.0–75.2% suppression).Conclusions/SignificanceOur study, the first open-field pilot implementation ofWolbachiaIIT-SIT in Mexico and Latin-America, confirms that inundative male releases can significantly reduce natural populations ofAe.aegypti. More importantly, we present successful pilot results of the integration ofWolbachiaIIT-SIT within a IVM plan implemented by Ministry of Health personnel.

We evaluated the efficacy of aerial ultra-low-volume (ULV) insecticide spraying in field bioassays with caged

The World Health Organization (WHO) has recently recommended indoor residual spraying (IRS) as part of a vector control strategy to combat Aedes-borne diseases, including dengue, chikungunya, and Zika viruses. Hand compression sprayers have been used in malaria prevention and control programs worldwide since the 1950s and are a standard for IRS application. However, there are technological advances that should be considered to improve IRS application (e.g., flow-control valves, rechargeable-battery equipment, reduced-drift nozzles, etc.), particularly if interventions are performed in urban areas to target Aedes aegypti. Using WHO guidelines, we contrasted technical characteristics of potential IRS equipment including hand compression sprayers (Hudson X-pert, Goizper IK Vector Control Super), rechargeable-battery sprayers (Solo 416, Birchmeier REC 15ABZ, Hudson NeverPump), and motorized sprayers (Honda WJR 2525, Kawashima AK35GX). Measurements included flow rate, droplet size, battery/fuel life, and technical/physical characteristics. Flow rate, the most important parameter, of the Hudson X-pert was stabilized at 550 ml/min by the use of a control flow valve (CFV). The IK Vector Control Super had integrated CFVs and produced a similar flow as the Hudson X-pert. Rechargeable-battery equipment provided consistent flow as well as negligible noise. Motorized sprayers also produced consistent flow, but their weight, high noise pollution when used indoors, and high engine temperature made them highly unpleasant for technicians. We identify alternatives to the more traditional hand compression Hudson X-pert sprayer with technical and operational considerations for performing IRS.

Fitting long-lasting insecticidal nets (LLIN) as screens on doors/windows has a significant impact on indoor-adult Aedes aegypti (L.), with entomological reductions measured in a previous study being significant for up to 2 yr post-installation, even in the presence of pyrethroid-resistant Aedes populations. To better understand the mode of LLIN protection, bioassays were performed to evaluate the effects of field deployment (0, 6, and 12 mo) and damage type (none, central, lateral, and multiple) on LLIN efficacy. Contact bioassays confirmed that LLIN residual activity (median knockdown time, in minutes, or MKDT) decreased significantly over time: 6.95 (95% confidence interval [CI]: 5.32–8.58) to 9.24 (95% CI: 8.69–9.79) MKDT at 0- and 12-mo age, respectively, using a pyrethroid-susceptible Aedes strain. Tunnel tests (exposing human forearm for 40 min as attractant) showed that deployment time affected negatively Aedes passage inhibition from 54.9% (95% CI: 43.5–66.2) at 0 mo to 35.7% (95% CI: 16.3–55.1) at 12 mo and blood-feeding inhibition from 65.2% (95% CI: 54.2–76.2) to 48.9% (95% CI: 26.4–71.3), respectively; both the passage/blood-feeding inhibition increased by a factor of 1.8–2.9 on LLINs with multiple and central damages compared with nets with lateral damage. Mosquito mortality was 74.6% (95% CI: 65.3–83.9) at 0 mo, 72.3% (95% CI: 64.1–80.5) at 6 mo, and 59% (95% CI: 46.7–71.3) at 12 mo. Despite the LLIN physical integrity could be compromised over time, we demonstrate that the remaining chemical effect after field conditions would still contribute to killing/repelling mosquitoes.

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.

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.

Objetivo. Determinar la eficacia de repelentes comerciales disponibles en Yucatán contra el mosquito Aedes aegypti, vector del dengue, Chikungunya y Zika. Material y métodos. Se determinó el tiempo de protección con base en el protocolo WHO/CTD/WHOPES/IC y la NOM-032- SSA2-2014. Resultados. Sólo el repelente con DEET (N, N-dietil-3-metilbenzamida) al 25% cumplió con la protección recomendada. La eficacia fue directamente proporcional a la concentración del DEET; aquéllos con componentes botánicos fueron poco o nada protectores. Conclusiones. Los resultados muestran que los repelentes con DEET proveen protección contra Ae. Aegypti; los repelentes botánicos, incluyendo las pulseras impregnadas, ofrecen nula protección. DOI: http://dx.doi.org/10.21149/spm.v58i4.8030

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.

Dengue is one of the most important vector-borne diseases worldwide and is a public health problem in Mexico. Most programs in dengue endemic countries rely on insecticides for Aedes control. In Mexico, pyrethroid insecticides (mainly permethrin and deltamethrin) have been extensively used over a decade as adulticides and represented a strong selection for insecticide resistance for dengue vectors in several parts of the country. We studied the type, frequency and distribution of insecticide resistance mechanisms in Aedes aegypti from six municipalities in the state of Guerrero selected on the basis of historically intense chemical control and a high risk for dengue transmission. Ae. aegypti eggs were collected from October 2009 to January 2010 using ovitraps. F1 adults, emerged from these collections, were exposed to permethrin, deltamethrin and DDT in WHO diagnostic tests and showed high resistance levels to both pyrethroids and DDT. This was consistent with the presence of increased metabolic enzyme activities and target site insensitivity due to kdr mutations. Biochemical assays showed elevated esterase and glutathione S-transferase activities in the six municipalities. The V1016I kdr mutation on the IIS6 domain of the sodium channel gene was present in an overall frequency of 0.80. A second mutation, F1534C on the IIIS6 domain of the same gene was also detected, being the first report of this mutation in Guerrero. The multiple resistance mechanisms present in Ae. aegypti from Guerrero state represent a warning for the efficacy of the pyrethroid usage and consequently for the success of the dengue control program.

OBJETIVO: Determinar la importancia relativa de los tipos de criadero de Aedes aegypti para proponer intervenciones de control enfocadas en Morelos. MATERIAL Y METODOS: Se realizaron muestreos transversales con colecta de pupas en Cuautla, Jojutla y Tlaquiltenango en temporada de sequia (1713 casas) y lluvias (1677) del ano 2008. La importancia relativa de cada tipo de criadero se determino por su contribucion (%) a la produccion pupal total por localidad. RESULTADOS: En Cuautla, la mayoria de pupas se recolectaron en temporada de sequia de tanques/pilas (48.5%), tambos y botes/cubetas (15% c/u); en lluvias, los diversos chicos (21.3%), botes/cubetas (19.3%) y macetas/macetones (12.9%) fueron mas productivos. En Jojutla y Tlaquiltenango, 97% de las pupas se colectaron de macetas/macetones en secas; durante las lluvias la mayoria de pupas se recolectaron de diversos chicos (26.3%), trastes de cocina/lavado (13.9%), botes/cubetas (12.9%) y macetas/macetones (12.7%), respectivamente. CONCLUSION: La prevencion y control del vector del dengue deben basarse en este tipo de evidencias para focalizar las acciones sobre los criaderos mas productivos.

We assessed the efficacy of commercial repellents available in Yucatan against Aedes aegypti, vector of dengue, Chikungunya and Zika.Protection time was determined based on WHO/CTD/ WHOPES/IC y la NOM-032-SSA2-20I4.Products with DEET (N, N-diethyl-3-methylbenzamide) at 25% met the recommended protection. Efficacy was directly proportional to the concentration of DEET; botanicals repellents resulted no protective.Repellents with DEET provided more protection against Ae. aegypti and botanical repellents, including impregnated wristbands, provided no protection.

Background \ud Long-lasting insecticidal net screens (LLIS) fitted to domestic windows and doors in combination with targeted treatment (TT) of the most productive Aedes aegypti breeding sites were evaluated for their impact on dengue vector indices in a cluster-randomised trial in Mexico between 2011 and 2013.\ud \ud Methods \ud Sequentially over 2 years, LLIS and TT were deployed in 10 treatment clusters (100 houses/cluster) and followed up over 24 months. Cross-sectional surveys quantified infestations of adult mosquitoes, immature stages at baseline (pre-intervention) and in four post-intervention samples at 6-monthly intervals. Identical surveys were carried out in 10 control clusters that received no treatment.\ud \ud Results \ud LLIS clusters had significantly lower infestations compared to control clusters at 5 and 12 months after installation, as measured by adult (male and female) and pupal-based vector indices. After addition of TT to the intervention houses in intervention clusters, indices remained significantly lower in the treated clusters until 18 (immature and adult stage indices) and 24 months (adult indices only) post-intervention.\ud \ud Conclusions \ud These safe, simple affordable vector control tools were well-accepted by study participants and are potentially suitable in many regions at risk from dengue worldwide.

Dengue prevention efforts rely on control of virus vectors. We investigated use of insecticide-treated screens permanently affixed to windows and doors in Mexico and found that the screens significantly reduced infestations of Aedes aegypti mosquitoes in treated houses. Our findings demonstrate the value of this method for dengue virus vector control.

We carried out dengue virus surveillance in Aedes aegypti populations from 47 neighborhoods of Acapulco during the rainy season of 2011 following a standard national protocol and as an improvement of the entomological surveillance of the Mexican Dengue Control Program. A total of 4,146 Ae. aegypti adults collected indoors and/or emerged from eggs, larvae, or pupae from households with dengue reports (probable or confirmed cases), were grouped into pools and processed using a standardized serotype-specific 4-plex real-time reverse transcriptase-polymerase chain reaction assay. Overall, only 2 (0.9%) of 226 pools of Ae. aegypti adults (1 pool of adults emerged from field-collected larvae, and another of indoor-collected adults) were positive for dengue virus 1 (DENV-1). This is appears to be the 1st report of evidence on the vertical and transovarial transmission of DENV-1 in field-caught Ae. aegypti in Mexico.

Background Dengue, recognized by the WHO as the most important mosquito-borne viral disease in the world, is a growing problem. Currently, the only effective way of preventing dengue is vector control. Standard methods have shown limited effect, and there have been calls to develop new integrated vector management approaches. One novel tool, protecting houses with long lasting insecticidal screens on doors and windows, is being trialled in a cluster randomised controlled trial by a joint UADY/WHO TDR/IDRC study in various districts of Acapulco, Mexico, with exceptionally high levels of crime and insecurity. This study investigated the community’s perspectives of long lasting insecticidal screens on doors and windows in homes and in schools, in order to ascertain their acceptability, to identify challenges to further implementation and opportunities for future improvements. Methods This was a sequential mixed-methods study. The quantitative arm contained a satisfaction survey administered to 288 houses that had received the intervention examining their perspectives of both the intervention and dengue prevention in general. The qualitative arm consisted of Focus Group Discussions (FGDs) with those who had accepted the intervention and key informant interviews with: schoolteachers to discuss the use of the screens in schools, program staff, and community members who had refused the intervention. Results Overall satisfaction and acceptance of the screens was very high, with only some operational and technical complaints relating to screen fragility and the installation process. However, the wider social context of urban violence and insecurity was a major barrier to screen acceptance. Lack of information dissemination and community collaboration were identified as project weaknesses. Conclusions The screens are widely accepted by the population, but the project implementation could be improved by reassuring the community of its legitimacy in the context of insecurity. More community engagement and better information sharing structures are needed. The screens could be a major new dengue prevention tool suitable for widespread use, if further research supports their entomological and epidemiological effectiveness and their acceptability in different social and environmental contexts. Further research is needed looking at the impact of insecurity of dengue prevention programmes. Electronic supplementary material The online version of this article (doi:10.1186/1471-2458-14-846) contains supplementary material, which is available to authorized users.

We compared the number of adult Aedes aegypti emerging from subsurface catch basins located in the streets against the number of pupae (as a proxy of adults emerging) from the entire container larval habitats found at residential premises within 1 ha of a neighborhood in the Mexican city of Merida during 8 days in the dry season of 2012. Aedes aegypti adults were collected from 60% of the subsurface catch basins. They produced 12 adults/day/ha (95% confidence interval [CI], 6.4 to 17.9), 5 females (95% CI, 2.1 to 7.7), and 7 males (95% CI, 3.8 to 10.7). In contrast, only 7 containers holding water were identified in 30 premises inspected, 1 bucket was positive for Ae. aegypti larvae, but no pupae-positive containers were found. No other mosquito species were found. This study revealed the importance of this type of nonresidential and subterranean aquatic habitat for Ae. aegypti adult production in this neighborhood of Merida during the dry season.

We report the collection of Aedes aegypti, Culex quinquefasciatus, Cx. interrogator, Cx. thriambus, Cx. coronator, and Cx. salinarius larvae from storm sewers within an endemic area for dengue transmission in Merida, Mexico, during the rainy season of 2011. This is the first record of the dengue vector Ae. aegypti breeding in storm sewers in the southeast of Mexico.

Aedes albopictus, Culex corniger, Cx. restuans, and Toxorhynchites theobaldi are reported for the first time for the Mexican State of Morelos. The updated list of species reported is also presented.

To determine the relative importance of Aedes aegyti breeding sites for potential targeted dengue control interventions in Morelos.Cross-sectional entomological surveys were conducted and collection of Ae. aegypti pupae was taken from all water-holding containers in Cuautla, Jojutla and Tlaquiltenango during dry (1,713 households) and rainy (1,677) seasons in 2008. Relative importance of different types of breeding sites was determined by the contribution (%) to total pupae production within each locality.In Cuautla most pupae during the dry season were found in wash basins (48.5%), tanks and buckets/pots (15% each); during the rainy season, diverse small items (21.3%), buckets/pots (19.3%) and plant pots (12.9%) were more productive. In Jojutla and Tlaquiltenango, 97% of all pupae was found in plant pots during the dry season; during the rainy season diverse small items (26.3%), washing/cooking utensils (13.9%), buckets/pots (12.9%) and plant pots (12.7%) were significant.Prevention and control of the dengue vector should be based on this kind of evidence to target the most productive breeding-sites.

Background Dengue is a major public health problem in Mexico, where the use of chemical insecticides to control the principal dengue vector, Aedes aegypti, is widespread. Resistance to insecticides has been reported in multiple sites, and the frequency of kdr mutations associated with pyrethroid resistance has increased rapidly in recent years. In the present study, we characterized patterns of insecticide resistance in Ae. aegypti populations in five small towns surrounding the city of Merida, Mexico. Methods A cross-sectional, entomological survey was performed between June and August 2013 in 250 houses in each of the five towns. Indoor resting adult mosquitoes were collected in all houses and four ovitraps were placed in each study block. CDC bottle bioassays were conducted using F0-F2 individuals reared from the ovitraps and kdr allele (Ile1016 and Cys1534) frequencies were determined. Results High, but varying, levels of resistance to chorpyrifos-ethyl was detected in all study towns, complete susceptibility to bendiocarb in all except one town, and variations in resistance to deltamethrin between towns, ranging from 63–88 % mortality. Significant associations were detected between deltamethrin resistance and the presence of both kdr alleles. Phenotypic resistance was highly predictive of the presence of both alleles, however, not all mosquitoes containing a mutant allele were phenotypically resistant. An analysis of genotypic differentiation (exact G test) between the five towns based on the adult female Ae. aegypti collected from inside houses showed highly significant differences (p