Mortality caused by passive resin transfluthrin diffusers (∼5 mg AI per 24 h release rate) suspended in small 2-person tents was measured for colony-reared sentinel pyrethroid susceptible Aedes aegypti and Culex quinquefasciatus female mosquitoes, as well as a pyrethroid-resistant strain of Aedes aegypti, in a USA military field camp scenario. Mortality effects were investigated for impact by factors such as sentinel cage location (inside tent, tent doorway and outside tent), exposure time (15, 30, 45 and 60 min), and environmental temperature (°C), all of which were examined over an 8-week period. Analyses determined there was a significant interaction between mosquito strain and transfluthrin susceptibility, with the two susceptible strains experiencing significantly greater mean mortality than the resistant Ae. aegypti strain. Significant differences were likewise observed between the mosquito strains over the 8-week study period, where study week and temperature were both positively correlated with an increase in observed mean mosquito mortality. Mosquito proximity to the transfluthrin diffusers was also influenced by week and showed that sentinel cage placement in the environment demonstrates different mortality measurements, depending on the environmental conditions. The length of exposure to transfluthrin, however, was determined to not significantly impact transfluthrin efficacy on the examined mosquito strains, although increased exposure did result in increased susceptible strain mortality. These results suggest that transfluthrin is highly effective in causing mortality against susceptible Ae. aegypti and Cx. quinquefasciatus mosquitoes under field conditions but is minimally effective against pyrethroid-resistant Ae. aegypti mosquitoes. Transfluthrin-infused devices are influenced by environmental factors that can combine to impact mosquito mortality in the field.

The prevention of vector-borne disease to protect the health and readiness of United States forces in the field continues to be a high priority for the US Department of Defense. Previous studies have demonstrated that the risk of human contact with disease-vector mosquitoes and other biting flies can be reduced by applying an insecticide to perimeters of military materials such as camouflage netting or HESCO blast protection wall geotextile already in place around troops in the field. In this study we investigated whether residual pesticide efficacy will persist in the presence of earth fill that is required for operational use of HESCOs, using a warm temperate field site in north Florida. Results from laboratory bioassays measuring mosquito mortality and field collections of natural mosquito populations indicated superior efficacy and greater longevity of pesticide treated geotextile exposed to soil fill. These findings not only support immediate implementation of this technique in US military field scenarios, but also provide evidence that HESCO technology currently used in natural disaster flood control could be leveraged to protect civilian personnel from emerging floodwater mosquitoes.

The control of such human diseases as dengue, Zika, and chikungunya relies on the control of their vector, the Aedes aegypti mosquito, because there is no prevention. Control of mosquito vectors can rely on chemicals applied to the immature and adult stages, which can contribute to the mortality of non-targets and more importantly, lead to insecticide resistance in the vector. The sterile insect technique (SIT) is a method of controlling populations of pests through the release of sterilized adult males that mate with wild females to produce non-viable offspring. This paper describes the process of producing sterile males for use in an operational SIT program for the control of Aedes aegypti mosquitoes. Outlined here are the steps used in the program including rearing and maintaining a colony, separating male and female pupae, irradiating and marking adult males, and shipping Aedes aegypti males to the release site. Also discussed are procedural caveats, program limitations, and future objectives.

Recent experiments suggest spatial repellents may significantly reduce biting pressure from host-seeking riceland mosquitoes, such as Anopheles quadrimaculatus, in a warm-humid open-field habitat. However, little is known regarding efficacy of these formulations in partially enclosed spaces where US military personnel may be sheltered or concealed in an operational environment. In this study we investigated the capability of 3 spatial repellents-metofluthrin, linalool, and d-cis/trans allethrin-to reduce mosquito incursion into small open-top enclosures of US military camouflage netting. We found that metofluthrin was more effective in partially enclosed spaces compared with the open field, whereas both linalool and d-cis/trans allethrin provided superior protection in the open. These findings support strategic selection of spatial repellents depending on the environment immediately surrounding the host.

ABSTRACT The efficacy of CocoBear™ Larvicidal Oil and Aquatain® AMF Liquid Mosquito Film against larval and pupal Culex quinquefasciatus was compared (at maximum label field application rates) when applied to concrete troughs treated with composted cow manure. At 1 h posttreatment, CocoBear provided significantly greater reduction of mosquito larvae than Aquatain, but both products were equally effective in producing >97% control at 24 h. Each product provided >98% pupal reduction at 1 h posttreatment, with complete elimination of pupae from troughs at 24 h. CocoBear and Aquatain proved to be equally effective against Cx. quinquefasciatus immatures in organically enriched aquatic habitats.

Aedes aegypti is a prominent disease vector that is difficult to control through traditional integrated vector management due to its cryptic peridomestic immature-stage habitat and adult resting behavior, increasing resistance to pesticide formulations approved by the US Environmental Protection Agency, escalating deregistration of approved pesticides, and slow development of new effective chemical control measures. One novel method to control Ae. aegypti is the sterile insect technique (SIT) that leverages the mass release of irradiated (sterilized) males to overwhelm mate choice of natural populations of females. However, one potential liability of SIT is sex sorting errors prior to irradiation, resulting in accidental release of females. Our goal in this study was to test the extent to which irradiation affects female life-history parameters to assess the potential impacts of releasing irradiated females accidentally sorted with males. In this study, we determined that a radiation dose ≥30 Gy-a dose sufficient to sterilize males while preserving their mating competitiveness-may substantially impact longevity, bloodfeeding, oviposition, and egg hatch rate of female Ae. aegypti after being irradiated as pupae. These findings could reduce public concern for accidental release of females alongside irradiated males in an operational Ae. aegypti SIT control program.

We investigated the capability of transfluthrin on US military camouflage netting to reduce collections of tabanid biting flies in a warm-temperate field environment on the Gulf Coast of Florida. We found that transfluthrin significantly reduced collections of a variety of medically and veterinarily important tabanids inside protected areas by up to 96% upon initial treatment and up to 74% after 20 days posttreatment. These results suggest that transfluthrin could be an effective element in the US Department of Defense integrated pest management system and leveraged in civilian scenarios to protect livestock and humans from potential mechanical transmission of pathogens and disruption of activities caused by painful bites.

Residual pesticide treatment of US military materials such as camouflage netting and HESCO blast wall geotextile is an effective way to reduce biting pressure within protected perimeters. However, residual treatments eventually wane and require retreatment in situ, which may not be possible or practical in military scenarios. One solution is to install pesticide misting systems on treated perimeters, which may additively enhance residual treatments, and gradually retreat perimeter material as misted pesticide settles. In this investigation we show that pesticide misting can extend efficacy of residual treatments on HESCO geotextile against mosquitoes and sand flies in a hot-arid desert environment by 1-2 wk.

Standard residual pesticides applied to US military materials such as camouflage netting can reduce mosquito biting pressure in the field but may contribute to the evolution of resistance. However, residual applications of a spatial repellent such as transfluthrin could allow mosquitoes the opportunity to escape, only inducing mortality if insects linger, for example after becoming trapped in a treated tent. In this study we investigated the capability of transfluthrin on 2 types of US military material to reduce natural populations of disease vector mosquitoes in a cool-arid desert field environment in southern California. We found that transfluthrin could reduce Culex tarsalis incursion into protected areas by up to 100% upon initial treatment and up to 45% for at least 16 days posttreatment, showing that this compound could be an effective element in the US Department of Defense integrated vector management system appropriate for further study.

Efficacy of public health pesticides targeting nuisance and disease-vector insects such as mosquitoes, sand flies, and filth-breeding flies is not uniform across ecological zones. To best protect public and veterinary health from these insects, the environmental limitations of pesticides need to be investigated to inform effective use of the most appropriate pesticide formulations and techniques. We have developed a research program to evaluate combinations of pesticides, pesticide application equipment, and application techniques in hot-arid desert, hot-humid tropical, warm and cool temperate, and urban locations to derive pesticide use guidelines specific to target insect and environment. To these ends we designed a system of protocols to support efficient, cost-effective, portable, and standardized evaluation of a diverse range of pesticides and equipment across multiple environments. At the core of these protocols is the use of an array of small cages with colony-reared sentinel mosquitoes (adults and immatures) and sand flies (adults), strategically arranged in natural habitats and exposed to pesticide spray. Spatial and temporal patterns of pesticide efficacy are derived from percent mortality in sentinel cages, then mapped and visualized in a geographic information system. Maps of sentinel mortality data may be statistically compared to evaluate relative efficacy of a pesticide across multiple environments, or to study multiple pesticides in a single environment. Protocols may be modified to accommodate a variety of scenarios, including, for example, the vertical orientation of sentinels in canopy habitats or simultaneous testing of ground and aerial application methods.

Rift Valley fever virus (RVFV) is a mosquito-borne zoonotic arbovirus with important livestock and human health, and economic consequences across Africa and the Arabian Peninsula. Climate and vegetation monitoring guide RVFV forecasting models and early warning systems; however, these approaches make monthly predictions and a need exists to predict primary vector abundances at finer temporal scales. In Kenya, an important primary RVFV vector is the mosquito Aedes mcintoshi. We used a zero-inflated negative binomial regression and multimodel averaging approach with georeferenced Ae. mcintoshi mosquito counts and remotely sensed climate and topographic variables to predict where and when abundances would be high in Kenya and western Somalia. The data supported a positive effect on abundance of minimum wetness index values within 500 m of a sampling site, cumulative precipitation values 0 to 14 days prior to sampling, and elevated land surface temperature values ~3 weeks prior to sampling. The probability of structural zero counts of mosquitoes increased as percentage clay in the soil decreased. Weekly retrospective predictions for unsampled locations across the study area between 1 September and 25 January from 2002 to 2016 predicted high abundances prior to RVFV outbreaks in multiple foci during the 2006–2007 epizootic, except for two districts in Kenya. Additionally, model predictions supported the possibility of high Ae. mcintoshi abundances in Somalia, independent of Kenya. Model-predicted abundances were low during the 2015–2016 period when documented outbreaks did not occur, although several surveillance systems issued warnings. Model predictions prior to the 2018 RVFV outbreak indicated elevated abundances in Wajir County, Kenya, along the border with Somalia, but RVFV activity occurred west of the focus of predicted high Ae. mcintoshi abundances.

Rift Valley fever (RVF) is a zoonotic disease, that causes significant morbidity and mortality among ungulate livestock and humans in endemic regions. In East Africa, the causative agent of the disease is Rift Valley fever virus (RVFV) which is primarily transmitted by multiple mosquito species in Aedes and Mansonia genera during both epizootic and enzootic periods in a complex transmission cycle largely driven by environmental and climatic factors. However, recent RVFV activity in Uganda demonstrated the capability of the virus to spread into new regions through livestock movements, and underscored the need to develop effective mitigation strategies to reduce transmission and prevent spread among cattle populations. We simulated RVFV transmission among cows in 22 different locations of the Kabale District in Uganda using real world livestock data in a network-based model. This model considered livestock as a spatially explicit factor in different locations subjected to specific vector and environmental factors, and was configured to investigate and quantitatively evaluate the relative impacts of mosquito control, livestock movement, and diversity in cattle populations on the spread of the RVF epizootic. We concluded that cattle movement should be restricted for periods of high mosquito abundance to control epizootic spreading among locations during an RVF outbreak. Importantly, simulation results also showed that cattle populations with heterogeneous genetic diversity as crossbreeds were less susceptible to infection compared to homogenous cattle populations.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

ABSTRACT Preemptive treatment of dry habitats with an ultra-low volume (ULV) residual larvicide may be effective in an integrated vector management program to control populations of container-inhabiting Aedes mosquitoes, key vectors of Zika, dengue, and chikungunya viruses. We exposed dry, artificial containers placed in exposed and protected locations to Natular 2EC (spinosad) larvicide applied with a truck-mounted ULV sprayer in a simulated urban setting in North Florida, and later introduced water and Ae. aegypti or Ae. albopictus larvae to conduct bioassays. Up to 50% mortality was observed in bioassays, indicating further analysis of spinosad as a residual treatment application.

ABSTRACT We investigated the efficacy of a liquid larvicide, Natular 2EC® (spinosad), applied with ultra-low volume sprayer as a residual application during the dry season in southeastern Thailand against 4 medically important species—Aedes aegypti, Ae. albopictus, Anopheles dirus, and An. minimus. We found that this larvicide could be applied as a residual to dry areas known to collect water and potentially still be effective after rains or irrigation, which could increase the flexibility and efficiency of an integrated vector management program targeting these species. This investigation also demonstrated, for the 1st time, efficacy of spinosad against An. minimus and An. dirus.

United States military personnel deployed in austere hot-arid, temperate, and tropical conditions in remote locations worldwide rely on the Department of Defense (DoD) Pest Management System measures such as application of pesticide residual and ultra-low-volume or thermal fog treatments for protection from routine exposure to nuisance and disease-vector insects. Timed pesticide misting systems originally developed for residential outdoor use show promise as potential enhancements to standard DoD Pest Management System measures, in particular to protect perimeters surrounding US military field encampments. To investigate the capabilities of misting systems in remote locations in diverse environments, this technical engineering report describes the development of 2 key modifications to a commercial residential misting system to enable us to operate the system away from a standard power grid, and to safely operate the system with small volumes of a variety of pesticide misting formulations. The components needed to modify the commercial misting system include the following: 5,000-W inverter, 12-V 100 Ah battery, 24-gal tote, solar panel with charge controller, 18 in. (45.7 cm) 0-gauge wire with ½-in. (1.27-cm) eyebolt connectors, 5-gal container (wide mouthed), ½-in.-to-½-in. connector, ½-in. tubing, ⅜-in.-to-⅜-in. connector, ⅜-in. tubing, ¼-in.-to-¼-in. connector, ¼-in. flexible tubing, securable patio box (80 gal), 5-gal (18.9-liter) bucket (modified), and a canning funnel.

The invasive Aedes aegypti is an important disease vector increasing in frequency in hot-arid regions of the USA such as the Southwest. Within hot-arid surroundings this mosquito may be confined to peridomestic locations that tend to be cooler and humid, such as in lush, irrigated ornamental vegetation surrounding homes. However, to reach these habitat refugia, ultra-low volume (ULV) applications of insecticides targeting this mosquito must retain efficacy after being sprayed from the air or street where hot-arid conditions are prevalent. We investigated the efficacy of a biologically based larvicide, spinosad (Natular 2EC), applied as a ULV in a hot-arid environment targeting Aedes aegypti. We found that this pesticide is able to penetrate this environment and has the potential to act as a residual.

We conducted aerial fixed wing ultra low volume (ULV) spray trials with naled to investigate penetration of exposed and simulated cryptic habitat within opened buildings, partially sealed buildings, and outdoor locations targeting sentinel adult Aedes aegypti mosquitoes in north central Florida. Mortality was observed in open and closed buildings and outdoors, even in mosquitoes placed in cryptic habitats. Observations on the impact of building type, mosquito exposure method such as placement in cryptic habitat, and spray nozzle size on mosquito mortality are described and analyzed.