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Abstract While residual insecticide applications have the potential to decrease pathogen transmission by reducing the density of vectors and shifting the age structure of the adult mosquito population towards younger stages of development, this double entomological impact has not been documented for Aedes aegypti. Aedes collected from households enrolled in a cluster-randomized trial evaluating the epidemiological impact of targeted indoor residual spraying (TIRS) in Merida, Mexico, were dissected and their age structure characterized by the Polovodova combined with Christopher’s ovariole growth methods. In total, 813 females were dissected to characterize age structure at 1, 3, 6, and 9 months post-TIRS. Significant differences in the proportion of nulliparous Ae. aegypti females between the treatment groups was found at one-month post-TIRS (control: 35% vs. intervention: 59%), three months (20% vs. 49%) but not at six or nine months post-TIRS. TIRS significantly shiftted Ae. aegypti age structure towards younger stages and led to a non-linear reduction in survivorship compared to the control arm. Reduced survivorship also reduced the number of arbovirus transmitting females (those who survived the extrinsic incubation period). Our findings provide strong evidence of the full entomological impact of TIRS, with important implications for quantifying the epidemiological impact of vector control methods.

BackgroundIn 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 findingsA 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; PConclusions/significanceThis is the first randomized control trial to evaluate the entomological impact of any volatile pyrethroid on urban Ae. aegypti. It demonstrates that volatile pyrethroids can have a sustained impact on Ae. aegypti population densities and human-vector contact indoors. These effects occur despite the presence of pyrethroid-resistant alleles in the target population. Formulations like these may have considerable utility for public health vector control responses.

Abstract Background Mexico has experienced a significant reduction in malaria cases over the past two decades. Certification of localities as malaria-free areas (MFAs) has been proposed as a steppingstone before elimination is achieved throughout the country. The Mexican state of Quintana Roo is a candidate for MFA certification. Monitoring the status of insecticide susceptibility of major vectors is crucial for MFA certification. This study describes the susceptibility status of Anopheles albimanus, main malaria vector, from historically important malaria foci in Quintana Roo, using both phenotypic and genotypic approaches. Methods Adult mosquito collections were carried out at three localities: Palmar (Municipality of Othon P. Blanco), Buenavista (Bacalar) and Puerto Morelos (Puerto Morelos). Outdoor human-landing catches were performed by pairs of trained staff from 18:00 to 22:00 during 3-night periods at each locality during the rainy season of 2022. Wild-caught female mosquitoes were exposed to diagnostic doses of deltamethrin, permethrin, malathion, pirimiphos-methyl or bendiocarb using CDC bottle bioassays. Mortality was registered at the diagnostic time and recovery was assessed 24 h after exposure. Molecular analyses targeting the Voltage-Gated Sodium Channel (vgsc) gene and acetylcholinesterase (ace-1) gene were used to screen for target site polymorphisms. An SNP analysis was carried out to identify mutations at position 995 in the vgsc gene and at position 280 in the ace-1 gene. Results A total of 2828 anophelines were collected. The main species identified were Anopheles albimanus (82%) and Anopheles vestitipennis (16%). Mortalities in the CDC bottle bioassay ranged from 99% to 100% for all the insecticides and mosquito species. Sequence analysis was performed on 35 An. albimanus across the three localities; of those, 25 were analysed for vgsc and 10 for ace-1 mutations. All individuals showed wild type alleles. Conclusion The results demonstrated that An. albimanus populations from historical malaria foci in Quintana Roo are susceptible to the main insecticides used by the Ministry of Health.

Abstract 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.

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 (p>0,05). 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.

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.

BackgroundThere 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.MethodsWe 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".ResultsMean 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).ConclusionsChlorfenapyr 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.

Iron and copper chelation restricts Plasmodium growth in vitro and in mammalian hosts. The parasite alters metal homeostasis in red blood cells to its favor, for example metabolizing hemoglobin to hemozoin. Metal interactions with the mosquito have not, however, been studied. Here, we describe the metallomes of Anopheles albimanus and Aedes aegypti throughout their life cycle and following a blood meal. Consistent with previous reports, we found evidence of maternal iron deposition in embryos of Ae. aegypti, but less so in An. albimanus. Sodium, potassium, iron, and copper are present at higher concentrations during larval developmental stages. Two An. albimanus phenotypes that differ in their susceptibility to Plasmodium berghei infection were studied. The susceptible white stripe (ws) phenotype was named after a dorsal white stripe apparent during larval stages 3, 4, and pupae. During larval stage 3, ws larvae accumulate more iron and copper than the resistant brown stripe (bs) phenotype counterparts. A similar increase in copper and iron accumulation was also observed in the susceptible ws, but not in the resistant bs phenotype following P. berghei infection. Feeding ws mosquitoes with extracellular iron and copper chelators before and after receiving Plasmodium-infected blood protected from infection and simultaneously affected follicular development in the case of iron chelation. Unexpectedly, the application of the iron chelator to the bs strain reverted resistance to infection. Besides a drop in iron, iron-chelated bs mosquitoes experienced a concomitant loss of copper. Thus, the effect of metal chelation on P. berghei infectivity was strain-specific.