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Riffell, J. A.

Publications and source records attributed to Riffell, J. A..

13 recordsLinked to original sources

Sexually Dimorphic Visual Processing in the Yellow-Fever Mosquito, Aedes aegypti

Flying insects process motion across the visual scene to regulate their ground speed, correct course deviations, and behaviorally relevant stimuli such as avoiding predators or finding hosts. The yellow-fever mosquito Aedes aegypti provides a useful model for studying these visually guided behaviors, as both sexes use vision to track moving features. Approaching a moving target presents mosquitoes with a demanding visual problem: they must extract target motion from the changing visual scene while simultaneously controlling their own flight and responding to potentially dangerous movements. To understand this, we first studied free-flight responses to static and moving vertical targets in a wind tunnel system capable of projecting dynamic visual stimuli, and found that males and females are sexually dimorphic with respect to their responses to target motion. When the feature was static, both males and females advanced toward it during flight; however, when it oscillated laterally at 2 Hz, only females continued to pursue it. In a subsequent yaw-unrestrained magnetic tethering paradigm, we showed that this divergence cannot be simply explained by sex-specific differences in visuomotor tuning dynamics, while micro-CT reconstructions revealed only modest sexual differences in eye morphology and broadly similar patterns of estimated spatial acuity. Together, these results indicate that males retain the biomechanical capacity to track oscillating targets but actively suppress approach behavior during free flight, whereas females pursue salient features independently of target motion. This pattern aligns with sex-divergent visual ecologies because the risk of approaching a large moving feature too closely likely outweighs the benefits for males seeking mates, whereas for host-seeking females the reproductive necessity of blood feeding overrides this avoidance.

neuroscience↗

DNA barcoding and olfactory identification of attractive nectar sources for Aedes aegypti mosquitoes

Adult mosquitoes rely on plant-derived sugars for survival, reproduction, and flight, yet the plant taxa that mosquitoes encounter in nature and the odors that make those plants attractive remain poorly understood. Most studies of mosquito attraction to plant odors have focused on candidate plants selected a priori, rather than plants linked to field-collected mosquitoes. Here, we combined plant DNA barcoding, semi-field behavioral assays, and volatile profiling to identify field-associated plant resources relevant to Aedes aegypti. Plant DNA recovered from mosquitoes collected across three Florida counties revealed broad plant associations, including 90 genera spanning 37 families, with several taxa recurring across counties or appearing prominently within particular localities. Behavioral experiments in semi-field sticky-trap assays found that five field-associated plant taxa were significantly attractive relative to blank controls, indicating that taxa associated with mosquitoes in nature can also function as attractive cues under semi-field conditions. GC-MS analyses of headspace collections from 42 plant taxa detected 211 volatile compounds and revealed substantial variation in both total emission rate and odor composition among taxa. Although several compounds, including -pinene, limonene, 4-ethylacetophenone, 2-ethyl-1-hexanol, 4-ethylbenzaldehyde, and caryophyllene, were broadly distributed across plant groups, volatile profiles differed significantly among taxa and shared compounds often occurred at markedly different proportional abundances. The five behaviorally tested taxa likewise showed both overlap and divergence, sharing 17 compounds across all five taxa while differing in dominant constituents and total emissions. Together, these results show that Ae. aegypti interacts with a diverse set of plants in the field, and suggests nectar-seeking is shaped not simply by plant identity or total odor abundance, but by the composition and proportional structure of plant odors.

ecology↗

Olfactory preferences and chemical differences of fruit scents for Aedes aegypti mosquitoes

Feeding on the sugar of fruits and flowers is vital for mosquitoes and increases their lifespan, reproduction, and flight activity. Olfaction is a key sensory modality in mediating mosquito responses to sugar sources. Previous studies have demonstrated that natural nectar sources from fruits and flowers can vary in attractiveness to mosquitoes, with some sources preferred over others. However, how the attractiveness of different fruits relates to the chemical composition of their scent and the responses they evoke from the mosquitos peripheral olfactory system, is still not understood. In this study, we use closely related fruit species and their varieties to examine how changes in scent chemistry can influence the fruits attractiveness to Aedes aegypti mosquitoes and examine how the mosquitos olfactory responses (via electroantennogram recordings, or EAG) correlate with those differences. Our results show that mosquitoes are attracted to the scents of certain fruits (Mangifera indica, Prunus perspica, Psidium guajava, Musa acuminata), whereas others (Pyrus communis, Citrus limon) elicited responses not significantly different from the negative control. Chemical analyses of the scents showed that attractive fruits have distinct chemical profiles, and amongst closely related fruits, minor changes in the relative proportions of scent compounds can modify the attractiveness. These minor differences in the fruit scent were not reflected in the EAG responses, which showed similar responses to scents from different fruit species and closely related varieties. Experimentally altering the chemical proportion of a single compound in attractive scents caused a significant decrease in attraction to levels similar to the less attractive cultivars. Our results demonstrate that mosquitoes are sensitive to compound proportions in attractive odors, which have implications for the olfactory processing of complex odor sources, like those from plants or blood hosts. Summary StatementAedes aegypti mosquitoes show specific and selective fruit scent preferences related to differences in the proportion of compounds in the scent.

animal behavior and cognition↗

Spectral preferences of mosquitos are altered by odors

Vision underlies many important behaviors in insects generally and in mosquitos specifically. Mosquito vision plays a role in predator avoidance, mate finding, oviposition, locating vertebrate hosts, and vectoring disease. Recent work has shown that when sensitized to CO2, the visual responses of Aedes aegypti are wavelength-dependent, but little is known about how other olfactory stimuli can modulate visual responses. The visual cues associated with flowers, vertebrate hosts, or oviposition sites differs substantially and it is possible that odors might prime the mosquito visual system to respond to these different resources. To investigate the interplay of olfactory and visual cues, we adapted previously used wind tunnel bioassays to use quasi-monochromatic targets (390-740 nm) created with a novel LED synth. We coupled these visual targets with CO2 and the odors representative of vertebrate hosts, floral nectar or oviposition sites and assessed responses via 3D tracking of female mosquitos. When CO2 alone is present, we observe a lower preference for wavelengths in the green portion of the visible spectrum with a gradual increase as wavelengths moved towards the violet and red ends of the spectrum. However, when odors associated both with flowers and oviposition sites, we observed significant increases in mosquito preference for green (475-575 nm) stimuli. In contrast when vertebrate host odor was present, we saw increased preference for stimuli across the entire visible spectrum. These odor shifts in the mosquito spectral preferences suggest these preferences are not fixed and shift depending on behavioral context.

animal behavior and cognition↗

Multisensory integration in Anopheles mosquito swarms: The role of visual and acoustic information in mate tracking and collision avoidance

Male mosquitoes form aerial aggregations, known as swarms, to attract females and maximize their chances of finding a mate. Within these swarms, individuals must be able to recognize potential mates and navigate the dynamic social environment to successfully intercept a mating partner. Prior research has almost exclusively focused on the role of acoustic cues in mediating the ability of male mosquitoes to recognize and pursue flying females. However, the role of other sensory modalities in this behavior has not been explored. Moreover, how males avoid collisions with one another in the dense swarm while pursuing females remains poorly understood. In this study, we combined free-flight and tethered flight simulator experiments to demonstrate that swarming Anopheles coluzzii mosquitoes integrate visual and acoustic information to track conspecifics and avoid collisions. Our tethered experiments revealed that acoustic stimuli gated mosquito steering responses to visual objects simulating nearby mosquitoes, especially in males that exhibited attraction to visual objects in the presence of female flight tones. Additionally, we observed that visual cues alone could trigger changes in wingbeat amplitude and frequency of mosquitoes. These findings were corroborated by our free-flight experiments, which revealed that mosquitoes modulate their flight responses to nearby conspecifics in a similar manner to tethered animals, allowing for collision avoidance within swarms. Together, these results demonstrate that both males and females integrate multiple sensory inputs to mediate swarming behavior, and for males, the change in flight kinematics in response to multimodal cues allows them to simultaneously track females while avoiding collisions.

neuroscience↗

The evolution of borneol repellency in culicine mosquitoes

The use of essential oils derived from the camphor tree to repel mosquitoes is an ancient practice that originated in Southeast Asia and gradually spread to China and across Europe via the Maritime Silk Road. The olfactory mechanisms by which these oils elicit avoidance behavior are unclear. Here we show that plant bicyclic monoterpenoids and borneol specifically activate a neural pathway that originates in the orphan olfactory receptor neuron of the capitate peg sensillum in the maxillary palp, and projects to the mediodorsal glomerulus 3 in the antennal lobe. This neuron co-locates with two olfactory receptor neurons tuned to carbon dioxide and octenol that mediate human-host detection. We also confirm that borneol elicits repellency against human-seeking female mosquitoes. Understanding the functional role of the mosquito maxillary palp is essential to investigating olfactory signal integration and host-selection behavior.

evolutionary biology↗

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, to host and plant related odors under different environmental temperatures.

neuroscience↗

The olfactory gating of visual preferences to human skin and colors in mosquitoes

Mosquitoes track odors, locate hosts, and find mates visually. The color of a food resource, such as a flower or warm-blooded host, can be dominated by long wavelengths of the visible light spectrum (green to red for humans) and is likely important for object recognition and localization. However, little is known about the hues that attract mosquitoes or how odor affects mosquito visual search behaviors. We used a real-time 3D tracking system and wind tunnel that allowed careful control of the olfactory and visual environment to quantify the behavior of more than 1.3 million mosquito trajectories. We found that CO2 induces a strong attraction to specific hues, including those that humans perceive as cyan, orange, and red. Sensitivity to orange and red correlates with mosquitoes strong attraction to the color spectrum of human skin, which is dominated by these wavelengths. Attraction was eliminated by filtering the orange and red bands from the skin color spectrum and by introducing mutations targeting specific long-wavelength opsins or CO2 detection. Collectively, our results show that odor is critical for mosquitoes wavelength preferences and that the mosquito visual system is a promising target for inhibiting their attraction to human hosts.

neuroscience↗

Aedes aegypti vision-guided target recognition requires two redundant rhodopsins

Blood-feeding insects, such as the mosquito, Aedes (Ae.) aegypti, use multiple senses to seek out and bite humans [1, 2]. Upon exposure to CO2, the attention of female mosquitoes to potential human targets is greatly increased. Female mosquitoes use vision to assist them in honing in on hosts that may be up to 10 meters away [3-9]. Only after coming into close range do convective heat from skin and odors from volatile organic compounds come into play, allowing female mosquitoes to evaluate whether the object of interest might be a host [10, 11]. Here, using CRISPR/Cas9 we mutated the gene encoding Op1, which is the most abundant of the five rhodopsins expressed in the compound eyes of Ae. aegypti. Using a cage assay and a wind tunnel assay, we surprisingly found that elimination of op1 did not impair CO2-induced target seeking. We then mutated op2, which encodes the rhodopsin most similar to Op1, and also found that there was no impact on this behavior. Rather, mutation of both op1 and op2 was required to abolish vision-guided target recognition. In contrast to this defect, the double mutants still exhibited normal light attraction. By measuring the optomotor response, we found that the double mutants still recognized moving cues in their environment. In further support of the conclusion that the double mutant is not blind, we found that the animals retained an electrophysiological response to light, although it was diminished. This represents the first perturbation of vision in mosquitoes and indicates that hostseeking by Ae. aegypti depends on redundant rhodopsins.

neuroscience↗

Neuromodulation and Differential Learning Across Mosquito Species

Mosquitoes can learn to change their host-feeding behaviors, such as shifting activity times to avoid bednets or switching from biting animals to biting humans, leading to the transfer of zoonotic diseases. Dopamine is critical for insect learning, but its role in the antennal lobe remains unclear, and it is unknown whether different mosquito species learn the same odor cues. We assayed aversive olfactory learning and dopaminergic brain innervation in four mosquito species with different host preferences and report here that they differentially learn odors salient to their preferred host and innervation patterns vary across species. Using genetically-encoded GCaMP6s Aedes aegypti, we mapped odor-evoked antennal lobe activity and report that glomeruli tuned to \"learnable\" odors have significantly higher dopaminergic innervation. Changes in dopamine expression in the antennal lobes of diverse invertebrate species may be an evolutionary mechanism to adapt olfactory learning circuitry without changing brain structure and for mosquitoes an ability to adapt to other hosts when their preferred are no longer present.

neuroscience↗

Geosmin attracts Aedes aegypti mosquitoes to oviposition sites

Geosmin is one of the most recognizable and common microbial smells on the planet. Some insects, like mosquitoes, require microbial-rich environments for their progeny, whereas for other insects such microbes may prove dangerous. In the vinegar fly Drosophila melanogaster, geosmin is decoded in a remarkably precise fashion and induces aversion, presumably signaling the presence of harmful microbes. We have here investigated the effect of geosmin on the behavior of the yellow fever mosquito Aedes aegypti. In contrast to flies, geosmin is not aversive in mosquitoes but stimulates egg-laying site selection. Female mosquitoes could associate geosmin with microbes, including cyanobacteria consumed by larvae, who also find geosmin - as well as geosmin producing cyanobacteria - attractive. Using in vivo multiphoton imaging from mosquitoes with pan-neural expression of the calcium reporter GCaMP6s, we show that Ae. aegypti code geosmin in a similar fashion to flies, i.e. with extreme sensitivity and with a high degree of selectivity. We further demonstrate that geosmin can be used as bait under field conditions, and finally we show that geosmin, which is both expensive and difficult to obtain, can be substituted by beetroot peel extract, providing a cheap and viable mean of mosquito control and surveillance in developing countries.

neuroscience↗

Computational and experimental insights into the chemosensory navigation of Aedes aegypti mosquito larvae

Mosquitoes are prolific disease vectors that affect public health around the world. Although many studies have investigated search strategies used by host-seeking adult mosquitoes, little is known about larval search behavior. Larval behavior affects adult body size and fecundity, and thus the capacity of individual mosquitoes to find hosts and transmit disease. Understanding vector survival at all life stages is crucial for improving disease control. In this study we use experimental and computational methods to investigate the chemical ecology and search behavior of Aedes aegypti mosquito larvae. We show that larvae do not respond to several olfactory cues used by adult Ae. aegypti to assess larval habitat quality, but perceive microbial RNA as a potent foraging attractant. Second, we demonstrate that Ae. aegypti larvae use a strategy consistent with chemokinesis, rather than chemotaxis, to navigate chemical gradients. Using computational modeling, we further show that chemokinesis is more efficient than chemotaxis for avoiding repellents in ecologically relevant larval habitat sizes. Finally, we use experimental observations and computational analyses to demonstrate that larvae respond to starvation pressure by optimizing exploration behavior. Our results identify key characteristics of foraging behavior in a disease vector mosquito, including the identification of a surprising foraging attractant and an unusual behavioral mechanism for chemosensory preference. In addition to implications for better understanding and control of disease vectors, this work establishes mosquito larvae as a tractable model for chemosensory behavior and navigation.

animal behavior and cognition↗

Live calcium imaging of Aedes aegypti neuronal tissues reveals differential importance of chemosensory systems for life-history-specific foraging strategies

Aedes aegypti have a wide variety of sensory pathways that have supported success as a species as well as a highly competent vector of numerous debilitating infectious pathogens. Investigations into mosquito sensory systems and their effects on behavior are valuable resources for the advancement of mosquito control strategies. Numerous studies have elucidated key aspects of mosquito sensory systems, however there remains critical gaps within the field. In particular, compared to that of the adult form, there has been a lack of studies directed towards the immature life stages. Additionally, although numerous studies have pinpointed specific sensory receptors as well as relevant response behaviors, there has been a lack of studies able to monitor both concurrently. To begin filling aforementioned gaps, here we engineered Ae. aegypti to ubiquitously express a genetically encoded calcium indicator, GCaMP6s. Using this strain, combined with advanced confocal microscopy, we were able to simultaneously measure live stimulus-evoked calcium responses in both neuronal and muscle cells with a wide spatial range and resolution. Moreover, by coupling in vivo calcium imaging with behavioral assays we were able to gain functional insights into how stimulus-evoked neural and muscle activities are represented, modulated, and transformed in mosquito larvae enabling us to elucidate mosquito sensorimotor properties important for life-history-specific foraging strategies.\n\nSignificance StatementUnderstanding mosquito sensory systems and resulting behavior has been a major factor in the advancement of mosquito control innovations. Aedes aegypti larvae offer an effective life stage for further elucidating information on mosquito sensory systems. Due to their relatively simplified nervous system, mosquito larvae are ideal for studying neural signal transduction, coding, and behavior. Moreover, a better understanding of the larval sensory system may enable the development of novel control methodologies able to target mosquitoes before they reach a vector-competent stage. Here we engineer Ae. aegypti to ubiquitously express a genetically encoded calcium indicator, GCaMP6s and use this tool to observe links between sensorimotor responses and behavior by exploiting live calcium imaging as well as live tracking based behavioral assays.

neuroscience↗