Gamma-Irradiated Female Aedes aegypti Mosquitoes Exhibit Greater Susceptibility to Mayaro Virus.

Mayaro virus (MAYV) is an alphavirus endemic in many parts of Central and South America transmitted to humans by Aedes aegypti. Currently, there is no vaccine or treatment of Mayaro infection, and therefore it is essential to control transmission by reducing populations of Ae. aegypti. Unfortunately, Ae. aegypti are extremely difficult to control with traditional integrated vector management (IVM) because of factors such as growing resistance to a dwindling list of registered insecticides and cryptic immature and adult habitats. The sterile insect technique (SIT) by irradiation is gaining traction as a novel supplemental tool to IVM. The SIT is being used operationally to release large numbers of sterilized colony-reared male mosquitoes in an intervention area to overwhelm females in the natural population, eventually causing population decline because of high frequencies of unfertilized eggs. However, little is known about the effect of irradiation on vector competence for mosquito-borne viruses such as MAYV in females that may be accidentally reared, irradiated, and released alongside males. In this investigation, we exposed female Ae. aegypti pupae to radiation and evaluated vector competence after inoculation with MAYV. Infection and dissemination rates of irradiated (10 and 40 Gy) Ae. aegypti were higher than those of non-irradiated cohorts at 7 and 14 days after infection. Although these results indicate a need to maintain effective sex sorting prior to irradiation and release of Ae. aegypti, our results are consistent with several previous observations that vectorial capacity and vector competence are likely lower in irradiated than in nonirradiated females.

Point Protection with Transfluthrin against Musca domestica L. in a Semi-Field Enclosure.

House flies are notoriously difficult to control, owing to their tendency to live in close relationships with humans and their livestock, and their rapid development of resistance to chemical controls. With this in mind, we explored an alternative chemical control, a spatial repellent to deter Musca domestica L. from points we wanted to protect (i.e., a baited trap). Our results demonstrated that the synthetic spatial repellent, transfluthrin, is effective in preventing M. domestica adults from entering protected traps for both a susceptible strain (CAR21) and a field-acquired permethrin-resistant strain (WHF; 24 h LD50 resistance ratio of 150), comprising 22% and 28% of the total number of flies collected, respectively. These results are promising and demonstrate that transfluthrin can be an effective spatial repellent to protect points of interest where needed.

Aedes aegypti Controls AE. Aegypti: SIT and IIT-An Overview.

The sterile insect technique (SIT) and the incompatible insect technique (IIT) are emerging and potentially revolutionary tools for controlling Aedes aegypti (L.), a prominent worldwide mosquito vector threat to humans that is notoriously difficult to reduce or eliminate in intervention areas using traditional integrated vector management (IVM) approaches. Here we provide an overview of the discovery, development, and application of SIT and IIT to Ae. aegypti control, and innovations and advances in technology, including transgenics, that could elevate these techniques to a worldwide sustainable solution to Ae. aegypti when combined with other IVM practices.

Transfluthrin diffusers do not protect two-person US military tents from mosquitoes in open field and canopy warm-temperate habitats.

Spatial repellents are volatile or volatilized chemicals that may repel arthropod vectors in free space, preventing bites and reducing the potential for pathogen transmission. In a 21-week field study, we investigated the efficacy of passive transfluthrin-impregnated diffusers placed in two-person United States (US) military tents located in canopy and open field habitats in north Florida to prevent mosquitoes from entering. Mosquito collections with US Centers for Disease Control and Prevention traps baited with light and carbon dioxide were conducted weekly for weeks 0-4, every two weeks for weeks 5-10, and monthly for weeks 11-21. Our results demonstrated that these transfluthrin-impregnated devices did not function as spatial repellents as expected and did not create a mosquito-free zone of protection. Instead, we observed consistently higher collections of mosquitoes from tents with transfluthrin-impregnated diffusers, and higher rates of mosquito mortality in collections from tents with transfluthrin diffusers, compared to untreated control tents. Based on these findings we do not recommend the use of passive transfluthrin-impregnated diffusers for mosquito protection in two-person US military tents in warm-temperate environments similar to north Florida.

Does prior exposure to larvicides influence dengue virus susceptibility in Aedes aegypti (Diptera: Culicidae)?

Control of mosquito vector populations is primarily intended to reduce the transmission of pathogens they transmit. Use of chemical controls, such as larvicides, can have unforeseen consequences on adult traits if not applied properly. The consequences of under application of larvicides are little studied, specifically the impacts on pathogen infection and transmission by the vectors that survive exposure to larvicides. We compared vector susceptibility of Aedes aegypti (L.) for dengue virus, serotype 1 (DENV-1) previously exposed as larvae to an LC50 of different classes of insecticides as formulated larvicides. Larval exposure to insect growth regulators (methoprene and pyriproxyfen) significantly increased susceptibility to infection of DENV-1 in Ae. aegypti adults but did not alter disseminated infection or transmission. Larval exposure to temephos, spinosad, and Bti did not increase infection, disseminated infection, or transmission of DENV-1. Our findings describe a previously under observed phenomenon, the latent effects of select larvicides on mosquito vector susceptibility for arboviruses. These data suggest that there are unintended consequences of sublethal exposure to select larvicides that can influence susceptibility of Ae. aegypti to DENV infection, and indicates the need for further investigation of sublethal effects of insecticides on other aspects of mosquito biology, especially those parameters relevant to a mosquitoes ability to transmit arboviruses (life span, biting behavior, extrinsic incubation period).

Developing the radiation-based sterile insect technique (SIT) for controlling Aedes aegypti: identification of a sterilizing dose.

BACKGROUND: The sterile insect technique (SIT) is emerging as a tool to supplement traditional pesticide-based control of Aedes aegypti, a prominent mosquito vector of microbes that has increased the global burden of human morbidity and mortality over the past 50 years. SIT relies on rearing, sterilizing and releasing large numbers of male mosquitoes that will mate with fertile wild females, thus reducing production of offspring from the target population. In this study, we investigated the effects of ionizing radiation (gamma) on male and female survival, longevity, mating behavior, and sterility of Ae. aegypti in a dose-response design. This work is a first step towards developing an operational SIT field suppression program against Ae. aegypti in St. Augustine, Florida, USA. RESULTS: Exposing late-stage pupae to 50 Gy of radiation yielded 99% male sterility while maintaining similar survival of pupae to adult emergence, adult longevity and male mating competitiveness compared to unirradiated males. Females were completely sterilized at 30 Gy, and when females were dosed with 50 Gy, they had a lower incidence of blood-feeding than unirradiated females. CONCLUSION: Our work suggests that an ionizing radiation dose of 50 Gy should be used for future development of operational SIT in our program area because at this dose males are 99% sterile while maintaining mating competitiveness against unirradiated males. Furthermore, females that might be accidentally released with sterile males as a result of errors in sex sorting also are sterile and less likely to blood-feed than unirradiated females at our 50 Gy dose. © 2022 Society of Chemical Industry.

Lethal and Sublethal Concentrations of Formulated Larvicides Against Susceptible Aedes aegypti.

Chemical control of vectors depends on the effective application of formulated insecticides. In this study we evaluated formulated larvicides using a larval bioassay against susceptible Aedes aegypti. The estimated larvicide lethal concentrations for 50% mortality (LC50s) were 25.7 µg/liter (Natular 2EC), 3.13 µg/liter (Abate 4E), 0.43 µg/liter (Altosid), 0.03 µg/liter (Nyguard), and 500.6 ITU/liter (VectoBac12AS containing Bacillus thuringiensis israelensis). Sublethal effects were identified and documented from adults that survived exposure to these estimated LC50s (body size and sex proportion). We observed changes in net growth as measured by adult wing lengths. For those larvae exposed to estimated LC50s, the average size of adults was between 0.1% and 10.6% smaller for males and between 1.1% and 13.6% smaller for females compared to controls. Sex proportions varied between larvicides, but some were significantly different from the control, favoring greater survival of females than males.

Assessing transfluthrin mortality against Aedes aegypti and Culex quinquefasciatus inside and outside US military tents in a northern Florida environment.

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.

Spatial Repellents Protect Small Perimeters from Riceland Mosquitoes in a Warm-Humid Environment.

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.

Preparing Irradiated and Marked Male Aedes aegypti Mosquitoes for Release in an Operational Sterile Insect Technique Program.

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.

Gamma-Irradiation Reduces Survivorship, Feeding Behavior, and Oviposition of Female Aedes aegypti.

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.

Visualizing Efficacy of Pesticides Against Disease Vector Mosquitoes in the Field.

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.

Aerial ULV control of Aedes aegypti with naled (Dibrom) inside simulated rural village and urban cryptic habitats.

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.

Ultra-Low Volume Application of Spinosad (Natular 2EC) as a Residual in a Hot-Arid Environment Against Aedes aegypti.

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.

Ultra-Low Volume Application of Spinosad (Natular 2EC) Larvicide as a Residual in a Tropical Environment Against Aedes and Anopheles Species.

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.

TRUCK-MOUNTED NATULAR 2EC (SPINOSAD) ULV RESIDUAL TREATMENT IN A SIMULATED URBAN ENVIRONMENT TO CONTROL AEDES AEGYPTI AND AEDES ALBOPICTUS IN NORTH FLORIDA.

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.

Portable Battery Power and Small-Reservoir Modifications For Pesticide Misting Systems.

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, »-in.-to-»-in. connector, »-in. flexible tubing, securable patio box (80 gal), 5-gal (18.9-liter) bucket (modified), and a canning funnel.

Residual Pesticide On Hesco® Blast Protection Wall In Temperate Florida Habitat Effective Against Mosquitoes, Stable Flies, and Sand Flies.

United States military troops in the field are exposed to the environment and are thus at high risk for transmission of arboviruses, and degradation of mission from continual harassment from insects. Passive vector control, such as application of residual insecticides to US military materials common in the field such as tents and camouflage netting, has been shown to be effective and can contribute to a successful integrated vector management (IVM) plan in the field to reduce this risk. However, other common US military field materials have not been evaluated with residual pesticides. In this study we conducted the first known investigation of the efficacy and longevity of a residual pesticide containing λ-cyhalothrin applied to HESCO® blast protection wall geotextile. We exposed treated material to a temperate Florida environment and found that this treatment can be effective against sand flies, filth-breeding flies, and mosquitoes for at least 6 wk. This study provides evidence that residual treatment of this US military material may be leveraged as an IVM component to enhance the US Department of Defense pest management system.

Application Site and Mosquito Age Influences Malathion- and Permethrin-Induced Mortality in Culex quinquefasciatus (Diptera: Culicidae).

Concentrations of malathion and permethrin typical in droplets generated from ultra-low-volume and low-volume applications used to control mosquito populations were evaluated for efficacy against multiple-aged Culex quinquefasciatus Say (Diptera: Culicidae), using a topical bioassay. Although insecticide droplets will impinge on many exoskeletal body regions and a range of ages of mosquitoes in a population, traditional mosquito topical bioassays focus pesticide application to the mesothoracic pleural or dorsal regions across an average mosquito age (e.g., 3-7 d). Our results document nonuniform insecticide sensitivity across body regions at ages not previously assessed in mosquitoes (teneral and 14-d old). We expect our findings to influence the topical bioassay process, illustrating the difference in mosquito body regions and ages that ultimately may explain insecticide effectiveness wherever droplets impinge upon the mosquito body during field control applications.