The impact of non-lethal doses of pyriproxyfen on male and female Aedes albopictus reproductive fitness.

Introduction: Control of the mosquito Aedes albopictus is confounded by its behavior due to females preferring to oviposition in small natural and artificial containers that are often difficult to remove or treat with insecticides. Autodissemination strategies utilizing highly potent insect growth regulators (IGRs) have emerged as promising tools for the control of this container-inhabiting species. The intended goal of autodissemination approaches is to use mosquitoes to self-deliver an IGR to these cryptic oviposition locations. Previous studies have focused on the efficacy of these approaches to impact natural populations, but little focus has been placed on the impacts on mosquitoes when exposed to non-lethal doses of IGRs similar to the levels they would be exposed to with autodissemination approaches. Methods: In this study, the impact of non-lethal doses of pyriproxyfen (PPF) on the reproductive fitness of Ae. albopictus was investigated. Female and male Ae. albopictus mosquitoes were exposed to non-lethal doses of PPF and their fecundity and fertility were measured. To examine the impact of non-lethal doses of PPF, the expression of the ecdysone-regulated genes USP, HR3, and Vg, which are involved in vitellogenesis, was determined. Results: Our results demonstrated a significant reduction in female fecundity and in the blood feeding and egg hatching rates upon exposure to non-lethal doses of PPF. Oocyte development was also delayed in PPF-treated females. Furthermore, exposure to non-lethal doses of PPF altered the expression of the genes involved in vitellogenesis, indicating disruption of hormonal regulation. Interestingly, PPF exposure also reduced the sperm production in males, suggesting a potential semi-sterilization effect. Discussion: These findings suggest that non-lethal doses of PPF could enhance the efficacy of autodissemination approaches by impacting the reproductive fitness of both males and females. However, further research is needed to validate these laboratory findings in field settings and to assess their practical implications for vector control strategies.

Nectar Source and Pollinator Cross Contamination and Persistence of Pyriproxyfen Associated with the Use of Autodissemination Techniques for Mosquito Control.

Controlling container inhabiting mosquitoes such as Aedes aegypti and Ae. albopictus is often difficult because of the requirement to treat small and inaccessible cryptic sources of water where larvae are located. Autodissemination approaches based on the dissemination of insect growth regulators (IGRs) have been demonstrated as an effective means to treat these cryptic larval habitats and provide population control. Autodissemination approaches are attractive because they are based on the mosquitoes disseminating small amounts of IGRs compared to more traditional insecticide applications. While dissemination of small amounts of IGRs seems like an advantage, these approaches could lead to unintended transfer and effects on nontarget insect pollinators by delivering highly potent IGRs to nectar sources. Here we looked for the indirect and direct transfer of pyriproxyfen (PPF) to natural and artificial nectar sources and painted lady butterflies, Vanessa cardui, in semifield cages using the release of treated Ae. albopictus males or an autodissemination station. We also performed persistence tests of PPF in oviposition containers and natural and artificial nectar sources when exposed to laboratory and natural conditions. The data suggest that there is direct and indirect transfer to nectar sources and V. cardui associated with the use of autodissemination approaches. We discuss the results in the context of using autodissemination approaches for mosquito control and the potential risks these approaches may pose to nontarget insect pollinators.

Indirect transfer of pyriproxyfen to European honeybees via an autodissemination approach.

The frequency of arboviral disease epidemics is increasing and vector control remains the primary mechanism to limit arboviral transmission. Container inhabiting mosquitoes such as Aedes albopictus and Aedes aegypti are the primary vectors of dengue, chikungunya, and Zika viruses. Current vector control methods for these species are often ineffective, suggesting the need for novel control approaches. A proposed novel approach is autodissemination of insect growth regulators (IGRs). The advantage of autodissemination approaches is small amounts of active ingredients compared to traditional insecticide applications are used to impact mosquito populations. While the direct targeting of cryptic locations via autodissemination seems like a significant advantage over large scale applications of insecticides, this approach could actually affect nontarget organisms by delivering these highly potent long lasting growth inhibitors such as pyriproxyfen (PPF) to the exact locations that other beneficial insects visit, such as a nectar source. Here we tested the hypothesis that PPF treated male Ae. albopictus will contaminate nectar sources, which results in the indirect transfer of PPF to European honey bees (Apis mellifera). We performed bioassays, fluorescent imaging, and mass spectrometry on insect and artificial nectar source materials to examine for intra- and interspecific transfer of PPF. Data suggests there is direct transfer of PPF from Ae. albopictus PPF treated males and indirect transfer of PPF to A. mellifera from artificial nectar sources. In addition, we show a reduction in fecundity in Ae. albopictus and Drosophila melanogaster when exposed to sublethal doses of PPF. The observed transfer of PPF to A. mellifera suggests the need for further investigation of autodissemination approaches in a more field like setting to examine for risks to insect pollinators.