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Anopheles stephensiis a major vector of malaria in Asia and the Arabian Peninsula, and its recent invasion into Africa poses a significant threat to malaria control and elimination efforts on the continent. The mosquito is well-adapted to urban environments, and its presence in Africa could potentially lead to an increase in malaria transmission in cities. Most of the knowledge aboutAn. stephensiecology in Africa has been generated from studies conducted during the rainy season, when vectors are most abundant. Here, we provide evidence from the peak of the dry season in the city of Jigjiga, Ethiopia, and report the finding ofAn. stephensiimmature stages infesting predominantly water reservoirs made to support construction operations (in construction sites or associated with brick manufacturing businesses). Political and economic changes in Ethiopia (and particularly the Somali Region) have fueled an unprecedented construction boom since 2018 that, in our opinion, has been instrumental in the establishment, persistence and propagation ofAn. stephensivia the year-round availability of perennial larval habitats associated with construction. We argue that larval source management during the dry season may provide a unique opportunity for focused control ofAn. stephensiin Jigjiga and similar areas.

The Wolbachia-based approach is under evaluation as a control strategy against Aedes aegypti in Mexico. From 2017 to 2019, we performed a pilot study to evaluate an open-field mass-release of wAlbB-infected Ae. aegypti male mosquitoes, as part of an integrated vector management (IVM) plan led by the Ministry of Health in Mexico to suppress natural populations of Ae. aegypti in southern Mexico. Community engagement and social evaluation were part of the key activities conducted. Overall, results showed the positive benefits of this Wolbachia-based method in the reduction of Aedes mosquitoes (90%). Mosquito’s nuisance at bedtime and the increasing circulation of mosquitoes during the releasing days were reported as the negative perceptions of this method. Importantly, participants understood the difference between wild mosquitoes and those released as part of the project, as well as the importance of the IVM. A significant number of the population accepted and supported the project, and feedback was given to improve future mosquito-releasing activities. The social license was a key factor in the success of the intervention and should be part of innovative paradigms for mosquito-vector control strategies involving community engagement. We outline the Mexican experience of community engagement and social assessment in implementing a Wolbachia-based strategy.

House-screening (HS) using fixed-aluminium frames to reduce the risk of indoor infestation with Aedes aegypti mosquitoes as well as the risk of Aedes-transmitted diseases in communities living in endemic areas. However, the success of this approach has been hindered by the elevated cost of the aluminium-based materials as well as their professional installation, which cannot be afforded by people living under vulnerable conditions. Cost-saving strategies such as the use of low-cost materials including wood, PVC, and Velcro are within the list of HS options available and offered by HS businesses and/or Do-it-yourself (DIY) packages verbi gratia ready-made and ready-to-install mosquito-screens. Here, we evaluated the efficacy of low-cost frames constructed with different materials to protect against Ae. aegypti indoor infestation using experimental huts. The efficacy of protection in preventing female mosquito passing inside the huts of any of the options of frames was high (>93%) compared to the control with no-screen. People’s perceptions on the different materials showed the most “popular” alternative was the frame made of wood (62%). All the prototype-frames of HS made of different materials were effective at blocking Ae. aegypti entering-mosquitoes particularly, low-cost options like magnets and Velcro.

After the tropical storm Cristobal, we performed special adult entomological collections in the peri-domicile of 35 houses from 25 neighborhoods of Merida, 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.

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.

The implementation of new control strategies for Aedes aegypti (Ae. Aegpyti), a vector of dengue, chikungunya, and Zika viruses, requires communities to adopt specific behaviors to achieve the success of these innovations.We evaluated the effect of an educational intervention based on the Precede-Proceed Model (PPM) and the Diffusion of Innovations Theory (DIT) for the control and prevention of diseases transmitted by Ae. aegypti through release of male mosquitoes infected with Wolbachia bacteria in a suburban town in Yucatan, Mexico.From July 2019 to February 2020, a quasi-experimental study was carried out through an educational intervention (pre- and post-measurements) using quantitative-qualitative techniques, in a Yucatan suburban town where male mosquitoes with Wolbachia were released for the suppression of Ae. aegypti populations. Eleven educational workshops were attended by heads of household (n = 19) and schoolchildren (n = 11). Other 136 heads of household not attending the workshops received information individually.The educational intervention had a significant effect on the mean scores of the contributing and behavioral factors for adoption of innovation (p 0.05) in the pre- and post-intervention measurements.Innovative methods for the control and prevention of diseases related to Aedes aegypti can be strengthened through educational interventions supported by sound methodologies.Community health education, Aedes aegypti, Wolbachia, Mexico.

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.

Commercial aerosolized insecticides can be implemented as a community-based approach to targeted indoor residual spraying against Aedes aegypti, but their efficacy on pyrethroid-resistant mosquitoes has not yet been evaluated. Two commercial aerosolized products (H24 Poder Fulminante Ultra Eficaz®, carbamate, and Baygon Ultra Verde®, pyrethroid) were sprayed on common indoor surfaces e.g., cement, plywood, and cloth, and tested for their residual efficacy on susceptible and field-derived pyrethroid-resistant Ae. aegypti strains using the WHO cone bioassays. Overall, ≥80% 24-h mortality was observed for both products for at least 4 wk regardless of the mosquito strain or surface type used. H24 Poder Fulminante Ultra Eficaz showed the highest residual potency, sustaining >80% mortality for 7-wk posttreatment regardless of mosquito strain and surface type. For Baygon Ultra Verde, the mean mortality of female Ae. aegypti remained >80% for a shorter period (4–6 wk). Nonpyrethroid commercial aerosolized formulations can provide a lasting residual effect indoors compatible with the need for rapid and lasting mosquito control during outbreaks and may be suitable for community-based targeted indoor residual spraying.

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.

We evaluated the efficacy of bendiocarb (Ficam W® 80%) and pirimiphos-methyl (Actellic 300CS® 28.16%), applied to different surfaces potentially sprayable within houses during the application of a targeted indoor residual spraying (TIRS) against a field pyrethroid-resistant strain of Aedes aegypti. Bioassays with cones were performed on cement (walls), wood (doors), and textile (cloth) surfaces within typical houses in the Mexican city of Merida (n = 10). Optimal residual efficacy (>80% of mean mortality) of bendiocarb ranged from 3 months (cement) to 2 months (wood and textiles). Residual efficacy of pirimiphos-methyl ranged from 5 months (cement) to 2 months (wood and textiles). Both insecticides proved to be effective as adulticides against field Ae. aegypti and may be useful in mosquito control programs implementing TIRS with pyrethroid-resistant populations.

We evaluated the efficacy of aerial ultra-low-volume (ULV) insecticide spraying in field bioassays with caged Aedes aegypti in May 2017 in Puerto Vallarta, Jalisco, Mexico. The insecticides tested included an organophosphate (Mosquitocida UNO ULV) and a neonicotinoid–pyrethroid combination (Cielo). Two Ae. aegypti populations were evaluated: a field pyrethroid-resistant local strain (Puerto Vallarta) and an insecticide-susceptible laboratory strain (New Orleans). Knockdown after 1 h by both products was ≥97.0%, and mortality after 24 h was ≥98% for the susceptible laboratory strain. Knockdown of the local Puerto Vallarta field strain by both products after 1 h was ≥96.5%; and mosquito mortality after 24 h was also very high (≥98%). Meteorological conditions during this evaluation were favorable for aerial mosquito control and represented conditions that typically occur during adulticide space spray applications. Temperature oscillated between 24°C and 26°C with winds between 6 and 10 km/h. The majority of droplets met the droplet distribution criteria required for the insecticides. The evaluation demonstrated an acceptable performance of both products for Ae. aegypti control when applied undiluted at a rate of 199.4 ml/ha and 73.07 ml/ha for Mosquitocida UNO ULV and Cielo, respectively. The volume median diameter (VMD) droplet size was characterized at 31.3 μm and 37.3 μm, respectively.

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.

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: The prevalence of SARS-CoV-2 exposure in children during the global Covid-19 pandemic has been underestimated due to lack of testing and the relatively mild symptoms in adolescents. Understanding the exposure rates in the pediatric population is essential as children are the last to receive vaccines and can act as a source for SARS-CoV-2 mutants that may threaten vaccine escape. This study aims to quantify the prevalence of anti-SARS-CoV-2 serum antibodies in children in a major city in Mexico. Methods: We obtained socioeconomic information and blood samples from 1,005 children and tested the sera for anti-SARS-CoV-2 IgG and IgM antibodies using lateral flow immunochromatography. Findings: We found that 25·5% of children in our cohort were positive for anti-SARS-CoV-2 antibodies and there was no correlation between age and antibody prevalence. Children that lived with large families were statistically more likely to have antibodies against SARS-CoV-2. Spatial analyses identified two hotspots of high SARS-CoV-2 seroprevalence in the west of the city. Interpretation: These results indicate that a large urban population of children has been exposed to SARS-CoV-2 and that a correlating factor was the number of people within the child’s household. There is also a larger population of children that may be susceptible to future infection upon easing of social distancing measures. This is the first analysis of a large cohort of children in a Latin American country and the results may inform public health decisions and vaccination strategies. Funding Information: This work was supported by NIH grant U01AI148069 and received support from the Universidad Autonoma de Yucatan. Declaration of Interests: The authors declare no conflicts of interest Ethics Approval Statement: This study capitalized on a pediatric cohort established to evaluate the epidemiological impact of indoor residual spraying on arbovirus transmission and utilized serum stored after either participant written consent or verbal assent was attained.(7) All participants agreed to have their specimens kept for future use. All procedures for participant assent/consent, blood acquisition, and serum storage were approved by the Institutional Review Boards of Emory University (IRB00108666) and the Universidad Autonoma de Yucatan (CEI-05-2020).

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.

Additional file 2.

Additional file 3. SPIRIT 2013 Checklist: Recommended items to address in a clinical trial protocol and related documents.

Additional file 1. WHO Trial Registration Data Set (Version 1.3.1): checklist.

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