Developing methods for chilling, compacting, and sterilizing adult Aedes aegypti (Diptera: Culicidae) and comparing mating competitiveness between males sterilized as adults versus pupae for sterile male release.

The yellow fever mosquito, Aedes aegypti L., can transmit several pathogens responsible for human diseases. With insecticide resistance development becoming a concern, alternative control strategies are needed for Ae. aegypti. Sterile insect technique (SIT) is an increasingly popular option being explored. However, logistical issues in mass production and sterilization make it difficult to maintain a SIT program. Male mosquitoes are typically irradiated as pupae because this is the earliest developmental point at which females can be separated from males, but asynchrony in pupation and high variability in pupal responses to irradiation based on pupal age make it difficult to sterilize mass quantities of pupae on a regular schedule in a rearing facility. Young adult mosquitoes have wider windows for irradiation sterilization than pupae, which can allow facilities to have fixed schedules for irradiation. We produced a workflow for adult Ae. aegypti irradiation in a mosquito control district with an operational SIT program that currently irradiates pupae. The impacts of chilling, compaction, and radiation dose on survival were all assessed before combining them into a complete adult irradiation protocol. Males chilled up to 16 h prior to compaction and compacted to 100 males/cm3 during radiation resulted in low mortality. Males irradiated as adults had increased longevity and similar sterility compared to males irradiated as pupae. Additionally, males sterilized as adults were more sexually competitive than males sterilized as pupae. Thus, we have shown that irradiating adult males can be a viable option to increase the efficiency of this operational mosquito SIT program.

Quantification of permethrin resistance and kdr alleles in Florida strains of Aedes aegypti (L.) and Aedes albopictus (Skuse).

Recent outbreaks of locally transmitted dengue and Zika viruses in Florida have placed more emphasis on integrated vector management plans for Aedes aegypti (L.) and Aedes albopictus Skuse. Adulticiding, primarily with pyrethroids, is often employed for the immediate control of potentially arbovirus-infected mosquitoes during outbreak situations. While pyrethroid resistance is common in Ae. aegypti worldwide and testing is recommended by CDC and WHO, resistance to this class of products has not been widely examined or quantified in Florida. To address this information gap, we performed the first study to quantify both pyrethroid resistance and genetic markers of pyrethroid resistance in Ae. aegypti and Ae. albopictus strains in Florida. Using direct topical application to measure intrinsic toxicity, we examined 21 Ae. aegypti strains from 9 counties and found permethrin resistance (resistance ratio (RR) = 6-61-fold) in all strains when compared to the susceptible ORL1952 control strain. Permethrin resistance in five strains of Ae. albopictus was very low (RR<1.6) even when collected from the same containers producing resistant Ae. aegypti. Characterization of two sodium channel kdr alleles associated with pyrethroid-resistance showed widespread distribution in 62 strains of Ae. aegypti. The 1534 phenylalanine to cysteine (F1534C) single nucleotide polymorphism SNP was fixed or nearly fixed in all strains regardless of RR. We observed much more variation in the 1016 valine to isoleucine (V1016I) allele and observed that an increasing frequency of the homozygous V1016I allele correlates strongly with increased RR (Pearson corr = 0.905). In agreement with previous studies, we observed a very low frequency of three kdr genotypes, IIFF, VIFF, and IIFC. In this study, we provide a statewide examination of pyrethroid resistance, and demonstrate that permethrin resistance and the genetic markers for resistance are widely present in FL Ae. aegypti. Resistance testing should be included in an effective management program.

Evaluation of Pyriproxyfen Dissemination via Aedes albopictus From a Point-Source Larvicide Application in Northeast Florida.

The Asian tiger mosquito, Aedes albopictus , ranks among the most important vectors of dengue fever, Zika virus, and chikungunya virus. With no specific medications or vaccines available, vector control is the only way to combat these diseases. Autodissemination of the insect growth regulator pyriproxyfen (NyGuard®) from a point-source treatment was evaluated in field settings in northeast Florida. The objective of this study was to investigate the possibility of pyriproxyfen dissemination from a treatment site to nontreated oviposition sites via the skip oviposition behavior of Ae. albopictus. A spray application was made to a tire pile using a Stihl® SR 420 backpack sprayer. Autodissemination oviposition vases containing oak infusion water were positioned in groups of five at 25 to 400 m in 4 transects surrounding the tire pile. Two sets of 5 control vases containing oak infusion water were placed 1,500 m from the tire pile and oak infusion water samples were collected directly from the tire pile. Fifty milliliter samples were extracted from each vase weekly and preserved for pyriproxyfen residue analysis. All vases were analyzed at week 0 (4 h post-treatment), 1, 2, 4, and 6. Overall, there were no differences in pupal mortality between the control and autodissemination vases. The tire pile samples had significantly more mortality (P < 0.0001) out to 4 wk when compared to autodissemination and control vases.

Field evaluation of commercial off-the-shelf spatial repellents against the Asian tiger mosquito, Aedes Albopictus (Skuse), and the potential for use during deployment.

The Testing and Evaluation Department of the US Navy Entomology Center of Excellence (NECE), Naval Air Station, Jacksonville, Florida, is dedicated to the evaluation of novel equipment and vector control techniques to provide guidance on effective protection measures against human pathogens transmitted by blood-feeding arthropods. Personal protective measures (PPM), to include repellents, are part of a series of techniques that contribute toward reducing human-vector contact for globally and domestically deployed military forces. However, improper PPM use and limited availability has created vulnerabilities, causing troops to purchase spatial repellent products that are not approved by the Department of Defense. In order to ensure the most effective products are available, NECE has evaluated the spatial repellency response of Aedes albopictus (Skuse) to 4 commercial off-the-shelf (COTS) spatial repellents to provide technical guidance on proper use and effectiveness. The COTS products evaluated ThermaCELL, OFF! Clip On, Lentek Bite Shield, and Bug Button Mosquito Eliminator. A Biogents Sentinel (BGS) trap was placed in 5 locations with a spatial repellent device suspended at the level of the BGS trap opening over 4 of them (the fifth was control). Each trap catch was collected every 12 hours, at which time the spatial repellent device was rotated to the next position. Using this method, each spatial repellent device and control was rotated across each of the 5 locations a total of 6 times. Spatial repellent efficiency was evaluated by comparing the total number of mosquitoes collected in the BGS traps during a 12-hour period. The number of adult mosquitoes repelled by the ThermaCell spatial repellent was significantly more than other spatial repellents with the exception of OFF!. These data indicate that COTS products using repellent insecticide rather than botanicals are more effective at deterring Ae. albopictus from biting a host.

Temporal and storage effects on ultra-low volume droplets of insecticides collected on Teflon-coated slides.

Use of microscope slides is the most commonly used method to field-assess the droplet spectrum of ultra-low volume (ULV) sprays. Due to absence of analysis facilities during military deployments, slides must be stored and shipped, and the impact of delays in processing and storage conditions on droplets is unknown. This study was designed to evaluate the effect of storage temperatures and duration on droplets on Teflon-coated slides. Treatments included BVA-13 mineral oil, Kontrol 30-30 (30% permethrin), and Fyfanon (96.5% malathion), 2 slide wrapping techniques (proper and improper), and 2 storage temperatures (23 and 45 degrees C), replicated 6 times. The same areas of a slide were measured at different times for 56-58 days using the DropVision droplet measurement system. Regardless of the wrapping technique, droplets of BVA-13, Fyfanon, and Kontrol 30-30 on slides stored at 45 degrees C reduced significantly after 1, 2, and 1 day, respectively, but droplets on slides stored at 23 degrees C were not significantly affected. The results of this study may assist vector control professionals to accurately interpret the droplet size and help in the effective dispersal of ULV-applied insecticides.