Protocol for rearing and using mosquitoes for flight path tracking and behavioral characterization in wind tunnel bioassays.

Mosquito behavioral assays are an important component in vector research and control tool development. Here, we present a protocol for rearing Anopheles mosquitoes, performing host-seeking behavioral bioassays, and collecting 3D flight tracks in a large wind tunnel. We describe steps for setting up host-seeking landing assays, both as a non-choice and as a dual-choice assay, and analyzing flight tracks. This protocol can be applied in the research of several behavioral traits, including nectar seeking, resting, mating, and oviposition behavior. For complete details on the use and execution of this protocol, please refer to Carnaghi et al.1.

Visual and thermal stimuli modulate mosquito-host contact with implications for improving malaria vector control tools.

Malaria prevention relies on mosquito control interventions that use insecticides and exploit mosquito behavior. The rise of insecticide resistance and changing transmission dynamics urgently demand vector control innovation. To identify behavioral traits that could be incorporated into such tools, we investigated the flight and landing response of Anopheles coluzzii to human-like host cues. We show that landing rate is directly proportional to the surface area of thermal stimulus, whereas close-range orientation is modulated by both thermal and visual inputs. We modeled anopheline eye optics to theorize the distance at which visual targets can be detected under a range of conditions, and experimentally established mosquito preference for landing on larger targets, although landing density is greater on small targets. Target orientation does not affect landing rate; however, vertical targets can be resolved at greater distance than horizontal targets of the same size. Mosquito traps for vector control could be significantly enhanced by incorporating these features.

Effect of Temperature on Mosquito Olfaction.

Mosquitoes use a wide range of cues to find a host to feed on, eventually leading to the transmission of pathogens. Among them, olfactory cues (e.g., host-emitted odors, including CO2, and skin volatiles) play a central role in mediating host-seeking behaviors. While mosquito olfaction can be impacted by many factors, such as the physiological state of the insect (e.g., age, reproductive state), the impact of environmental temperature on the olfactory system remains unknown. In this study, we quantified the behavioral responses of Aedes aegypti mosquitoes, vectors of dengue, yellow fever, and Zika viruses, among other pathogens, to host and plant-related odors under different environmental temperatures.

Effect of temperature on mosquito olfaction.

Mosquitoes use a wide range of cues to find a host to feed on, eventually leading to the transmission of pathogens. Among them, olfactory cues ( e.g. , host emitted odors, including CO 2 , and skin volatiles) play a central role in mediating host seeking behaviors. While mosquito olfaction can be impacted by many factors, such as the physiological state of the insect ( e.g. , age, reproductive state), the impact of environmental temperature on the olfactory system remains unknown. In this study, we quantified the behavioral responses of Aedes aegypti mosquitoes, vectors of dengue, yellow fever and Zika viruses, to host and plant related odors under different environmental temperatures.