Mosquito Host-seeking Research Articles

Evaluation of Anopheline Diversity and Abundance across Outdoor Collection Schemes Utilizing CDC Light Traps in Nchelenge District, Zambia.

In the global fight against malaria, standard vector control methods such as indoor residual spraying (IRS) and insecticide-treated nets (ITNs) are intended to protect inside residential structures and sleeping spaces. However, these methods can still leave individuals vulnerable to residual transmission from vectors that they may be exposed to outdoors. Nchelenge District in northern Zambia experiences persistently high malaria transmission even with ITNs and IRS in place. However, very few studies have examined outdoor vector activity. To assess the diversity and abundance of outdoor foraging female anopheline mosquitoes, CDC light traps were used as proxy measures for mosquito host-seeking, set in three outdoor trapping schemes randomly assigned on different nights: (1) locations where people congregate at night outside of the house within the peri-domestic space, (2) animal pens or shelters, and (3) high-human-traffic areas, such as paths to latrines, where traps were baited with BG-Lure®. A total of 1087 total female anophelines were collected over a total of 74 trap nights. Anopheles funestus s.s. comprised the majority of the collection (86%), with An. gambiae s.s. (2%) and a highly diverse sampling of other anophelines (12%) making up the remainder. Plasmodium falciparum parasites were only detected in An. funestus (1%). No significant difference in species diversity or female anopheline abundance was detected between trapping schemes. Outdoor foraging anopheline mosquitoes, including a number of infectious An. funestus, may partially explain the difficulty of controlling malaria transmission in Nchelenge District, where vector control is only targeted indoors. BG-Lure® shows some promise as an alternative to human-baited landing catch collections in this resource-poor setting.

Phenotypic variation in biting behavior associated with differences in expression of olfactory genes in the vector mosquito, Aedes albopictus (Diptera: Culicidae).

We evaluated miRNA and mRNA expression differences in head tissues between avid-biting vs. reluctant-biting Aedes albopictus (Skuse) females from a single population over a 20-min timescale. We found no differences in miRNA expression between avid vs. reluctant biters, indicating that translational modulation of blood-feeding behavior occurs on a longer timescale than mRNA transcription. In contrast, we detected 19 differentially expressed mRNAs. Of the 19 differentially expressed genes at the mRNA level between avid-biting vs. reluctant-biting A. albopictus, 9 are implicated in olfaction, consistent with the well-documented role of olfaction in mosquito host-seeking. Additionally, several of the genes that we identified as differentially expressed in association with phenotypic variation in biting behavior share similar functions with or are inferred orthologues of, genes associated with evolutionary variation in biting behaviors of Wyeomyia smithii (Coq.) and Culex pipiens (Lin.). A future goal is to determine whether these genes are involved in the evolutionary transition from a biting to a non-biting life history.

House Screening Reduces Exposure to Indoor Host-Seeking and Biting Malaria Vectors: Evidence from Rural South-East Zambia.

This study evaluated the impact of combining house screens with long-lasting insecticidal nets (LLINs) on mosquito host-seeking, resting, and biting behavior. Intervention houses received house screens and LLINs, while control houses received only LLINs. Centre for Disease Control light traps, pyrethrum spray collections and human landing catches were used to assess the densities of indoor and outdoor host-seeking, indoor resting, and biting behavior of malaria vectors in 15 sentinel houses per study arm per sampling method. The protective efficacy of screens and LLINs was estimated through entomological inoculation rates (EIRs). There were 68% fewer indoor host-seeking Anopheles funestus (RR = 0.32, 95% CI 0.20-0.51, p < 0.05) and 63% fewer An. arabiensis (RR = 0.37, 95% CI 0.22-0.61, p < 0.05) in screened houses than unscreened houses. There was a significantly higher indoor biting rate for unscreened houses (6.75 bites/person/h [b/p/h]) than for screened houses (0 b/p/h) (χ2 = 6.67, df = 1, p < 0.05). The estimated indoor EIR in unscreened houses was 2.91 infectious bites/person/six months, higher than that in screened houses (1.88 infectious bites/person/six months). Closing eaves and screening doors and windows has the potential to reduce indoor densities of malaria vectors and malaria transmission.

Wind Tunnels and Airflow-Driven Assays: Methods for Establishing the Cues and Orientation Mechanisms That Modulate Female Mosquito Attraction to Human Hosts.

Understanding how female mosquitoes find a prospective host is crucial to developing means that can interfere with this process. Many methods are available to researchers studying cues and orientation mechanisms that modulate female mosquito attraction to hosts. Behaviors that can be monitored with these assays include activation, taking flight, upwind flight along an odor plume (optomotor anemotaxis), close approach to the stimulus (including hovering), and landing. Video recording can three-dimensionally document flight tracks and can correlate overall distribution patterns and moment-to-moment movements with odor contact and the presence of nearby cues such as a visual target. Here, we introduce mosquito host-seeking behaviors and methods to study them: wind tunnels (which allow orientation in free-flight), airflow-driven assays (using either tethered mosquitoes or small assay chambers that permit flight but also often dictate walking orientation), and still-air assays (wherein in odor concentration and spatial distribution are the orientation cues). We also describe factors that affect the assays and provide assay design considerations.

Knockout of OR39 reveals redundancy in the olfactory pathway regulating the acquisition of host seeking in Anopheles coluzzii.

The attraction of anthropophilic mosquitoes to human host cues, such as body odour and carbon dioxide, gradually increases during adult maturation. This acquisition of host-seeking behaviour correlates with age-dependent changes in odorant receptor (OR) transcript abundance and sensitivity of olfactory sensory neurons (OSNs). One OR gene of the human malaria vector, Anopheles coluzzii, AcolOR39, is significantly downregulated in mature females, and a cognate ligand of AcolOR39, sulcatone, a major component of human emanations, mediates the observed behavioural inhibition of newly emerged (teneral) females to human body odour. Knockout of AcolOR39, using CRISPR-Cas9 mutagenesis, selectively abolished sulcatone detection in OSNs, housed in trichoid sensilla. However, knockout of AcolOR39 altered neither the response rate nor the flight behaviour of teneral females in a wind tunnel, indicating the involvement of other genes, and thus a redundancy, in regulating the acquisition of host seeking in mosquitoes.

Perceived mosquito nuisance in residential areas, varying in distance to the floodplain

Mosquitoes, which are known as carriers of infectious pathogens, can induce public health threats. Moreover, high numbers of host-seeking nuisance mosquitoes in residential areas could lower life quality. In this study, we analysed how the mosquito nuisance perception of residents differed in four residential areas in Germany. Residential areas varied in distance to the nearest floodplain and mosquito breeding sites within this floodplain. We then surveyed residents in these areas and recorded adult mosquito field data with one mosquito trap per area. Nuisance perception of residents intensified in the studied area closest to the floodplain, while in the residential area at the greatest distance to the floodplain perception was low. The highest abundance ofAedes spp. was collected in the trap closest to the floodplain. We discussed the reactions of residents to the presence of specificAedes spp. species andCulex pipiens complex and how the distance to the floodplain could shape mosquito species composition. In conclusion, we assumed that residents might respond to the mosquito species-specific biting and host-seeking behaviours. Consequently, nuisance perception of the residents might change according to the distance between the floodplain and residential area and mosquito species present in the residential areas.

Semi-field evaluation of a volatile transfluthrin-based intervention reveals efficacy as a spatial repellent and evidence of other modes of action.

Presently, the most common malaria control tools-i.e., long lasting insecticide-treated nets (LLINs) and indoor residual spraying (IRS)-are limited to targeting indoor biting and resting behaviors of Anopheles mosquito species. Few interventions are targeted towards malaria control in areas where transmission is driven or persists due to outdoor biting behaviors. This study investigated a volatile pyrethroid-based spatial repellent (VPSR) designed to bridge this gap and provide protection from mosquito bites in outdoor spaces. Southern Province, Zambia, is one such environment where outdoor biting is suspected to contribute to malaria transmission, where people are active in the evening in open-walled outdoor kitchens. This study assessed the VPSR in replica kitchens within a controlled semi-field environment. Endpoints included effects on mosquito host seeking, immediate and delayed mortality, deterrence, blood feeding inhibition, and fertility. Host-seeking was reduced by approximately 40% over the course of nightly releases in chambers containing VPSR devices. Mosquito behavior was not uniform throughout the night, and the modeled effect of the intervention was considerably higher when hourly catch rates were considered. These two observations highlight a limitation of this overnight semi-field design and consideration of mosquito circadian rhythms is recommended for future semi-field studies. Additionally, deterrence and immediate mortality were both observed in treatment chambers, with evidence of delayed mortality and a dose related response. These results demonstrate a primarily personal protective mode of action with possible positive and negative community effects. Further investigation into this primary mode of action will be conducted through a field trial of the same product in nearby communities.

Quantifying Aedes aegypti Host Odor Preference Using a Two-Port Olfactometer.

Blood-feeding mosquitoes are a leading threat to global public health-vectoring dangerous infections including Zika, dengue, and malaria. Mosquitoes identify and target hosts for blood meals by using visual, thermal, and chemical cues. Here we describe an assay for measuring odor-based host-preference behavior-that is, the preferential approach toward one host over another based on differences in the volatile compounds they emit. The assay can be adapted for use with diverse odor sources, from live animals and their breath to odor-scented sleeves with controlled amounts of CO2 Mosquitoes in this assay fly upwind to within 30 cm of the odor source and then enter a small trap. We therefore believe this assay best replicates medium- to short-range host-seeking, when females approach and are preparing to land on a host animal. We also find that relative response in a two-choice test shows less trial-to-trial variation than the absolute number of responsive mosquitoes, which appears more sensitive to exogenous factors such as rearing conditions. This assay has been used to better understand mosquito host-seeking decisions, which can provide fundamental insight into the brain and behavior as well as information useful for the design of novel vector control strategies.

Mosquito host-seeking diel rhythm and chemosensory gene expression is affected by age and Plasmodium stages

Malaria parasites can affect vector-related behaviours, increasing transmission success. Using Anopheles gambiae and Plasmodium falciparum, we consider the effect of interaction between infection stage and vector age on diel locomotion in response to human odour and the expression of antennal chemosensory genes. We identified age-dependent behavioural diel compartmentalisation by uninfected females post-blood meal. Infection disrupts overall and diel activity patterns compared with age-matched controls. In this study, mosquitoes carrying transmissible sporozoites were more active, shifting activity periods which corresponded with human host availability, in response to human odour. Older, uninfected, blood-fed females displayed reduced activity during their peak host-seeking period in response to human odour. Age- and infection stage-specific changes in odour-mediated locomotion coincide with altered transcript abundance of select chemosensory genes suggesting a possible molecular mechanism regulating the behaviour. We hypothesize that vector-related behaviours of female mosquitoes are altered by infection stage and further modulated by the age post-blood meal of the vector. Findings may have important implications for malaria transmission and disease dynamics.

L-lactic and 2-ketoglutaric Acids, Odors from Human Skin, Govern Attraction and Landing in Host-Seeking Female Aedes aegypti Mosquitoes

Aedes aegypti, presented with a source of L-lactic and 2-ketoglutaric acid in a wind-tunnel bioassay, takeoff, fly upwind, and land on the odorant substrate at rates comparable to those exhibited by mosquitoes presented with a skin-odor stimulus. Addition of carbon dioxide decreased takeoff latency but was not required to elicit upwind flight nor landings. Ketoglutaric acid, a recently identified component of human skin odor, combined with lactic acid elicits the full repertoire of mosquito host-seeking behaviors.

A volatile from the skin microbiota of flavivirus-infected hosts promotes mosquito attractiveness

The host-seeking activity of hematophagous arthropods is essential for arboviral transmission. Here, we demonstrate that mosquito-transmitted flaviviruses can manipulate host skin microbiota to produce a scent that attracts mosquitoes. We observed that Aedes mosquitoes preferred to seek and feed on mice infected by dengue and Zika viruses. Acetophenone, a volatile compound that is predominantly produced by the skin microbiota, was enriched in the volatiles from the infected hosts to potently stimulate mosquito olfaction for attractiveness. Of note, acetophenone emission was higher in dengue patients than in healthy people. Mechanistically, flaviviruses infection suppressed the expression of RELMα, an essential antimicrobial protein on host skin, thereby leading to the expansion of acetophenone-producing commensal bacteria and, consequently, a high acetophenone level. Given that RELMα can be specifically induced by a vitamin A derivative, the dietary administration of isotretinoin to flavivirus-infected animals interrupted flavivirus life cycle by reducing mosquito host-seeking activity, thus providing a strategy of arboviral control.

Mosquito Host Seeking in 3D Using a Versatile Climate-Controlled Wind Tunnel System.

Future anthropogenic climate change is predicted to impact sensory-driven behaviors. Building on recent improvements in computational power and tracking technology, we have developed a versatile climate-controlled wind tunnel system, in which to study the effect of climate parameters, including temperature, precipitation, and elevated greenhouse gas levels, on odor-mediated behaviors in insects. To establish a baseline for future studies, we here analyzed the host-seeking behavior of the major malaria vector mosquito, Anopheles gambiae sensu strico, to human odor and carbon dioxide (CO2), under tightly controlled climatic conditions, and isolated from potential background contamination by the presence of an experimenter. When presented with a combination of human foot odor and CO2 (case study I), mosquitoes engaged in faster crosswind flight, spent more time in the filamentous odor plume and targeted the odor source more successfully. In contrast, female An. gambiae s. s. presented with different concentrations of CO2 alone, did not display host-seeking behavior (case study II). These observations support previous findings on the role of human host-associated cues in host seeking and confirm the role of CO2 as a synergist, but not a host-seeking cue on its own. Future studies are aimed at investigating the effect of climate change on odor-mediated behavior in mosquitoes and other insects. Moreover, the system will be used to investigate detection and processing of olfactory information in various behavioral contexts, by providing a fine-scale analysis of flight behavior.

A minimal 3D model of mosquito flight behaviour around the human baited bed net

BackgroundAdvances in digitized video-tracking and behavioural analysis have enabled accurate recording and quantification of mosquito flight and host-seeking behaviours, facilitating development of individual (agent) based models at much finer spatial scales than previously possible.MethodsQuantified behavioural parameters were used to create a novel virtual testing model, capable of accurately simulating indoor flight behaviour by a virtual population of host-seeking mosquitoes as they interact with and respond to simulated stimuli from a human-occupied bed net. The model is described, including base mosquito behaviour, state transitions, environmental representation and host stimulus representation.ResultsIn the absence of a bed net and human host bait, flight distribution of the model population was relatively uniform throughout the arena. Introducing an unbaited untreated bed net induced a change in distribution with an increase in landing events on the net surface, predominantly on the sides of the net. Adding the presence of a simulated human bait dramatically impacted flight distribution patterns, exploratory foraging and, the number and distribution of landing positions on the net, which were determined largely by the orientation of the human within. The model replicates experimental results with free-flying living mosquitoes at human-occupied bed nets, where contact occurs predominantly on the top surface of the net. This accuracy is important as it quantifies exposure to the lethal insecticide residues that may be unique to the net roof (or theoretically any other surface). Number of net contacts and height of contacts decreased with increasing attractant dispersal noise.ConclusionsResults generated by the model are an accurate representation of actual mosquito behaviour recorded at and around a human-occupied bed net in untreated and insecticide-treated nets. This fine-grained model is highly flexible and has significant potential for in silico screening of novel bed net designs, potentially reducing time and cost and accelerating the deployment of new and more effective tools for protecting against malaria in sub-Saharan Africa.

Malaria hotspots explained from the perspective of ecological theory underlying insect foraging

Hotspots constitute the major reservoir for residual malaria transmission, with higher malaria incidence than neighbouring areas, and therefore, have the potential to form the cornerstone for successful intervention strategies. Detection of malaria hotspots is hampered by their heterogenous spatial distribution, and the laborious nature and low sensitivity of the current methods used to assess transmission intensity. We adopt ecological theory underlying foraging in herbivorous insects to vector mosquito host seeking and modelling of fine-scale landscape features at the village level. The overall effect of environmental variables on the density of indoor mosquitoes, sporozoite infected mosquitoes, and malaria incidence, was determined using generalized linear models. Spatial analyses were used to identify hotspots for malaria incidence, as well as malaria vector density and associated sporozoite prevalence. We identify household occupancy and location as the main predictors of vector density, entomological inoculation rate and malaria incidence. We propose that the use of conventional vector control and malaria interventions, integrated with their intensified application targeting predicted hotspots, can be used to reduce malaria incidence in endemic and residual malaria settings.

Dengue infection modulates locomotion and host seeking in Aedes aegypti.

Pathogens may manipulate their human and mosquito hosts to enhance disease transmission. Dengue, caused by four viral serotypes, is the fastest-growing transmissible disease globally resulting in 50–100 million infections annually. Transmission of the disease relies on the interaction between humans and the vector Aedes aegypti and is largely dependent on the odor-mediated host seeking of female mosquitoes. In this study, we use activity monitors to demonstrate that dengue virus-1 affects the locomotion and odor-mediated behavior of Ae. aegypti, reflecting the progression of infection within the mosquito. Mosquitoes 4–6 days post-infection increase locomotion, but do not alter their odor-driven host-seeking response. In contrast, females 14–16 days post-infection are less active, yet more sensitive to human odors as assessed by behavioral and electrophysiological assays. Such an increase in physiological and behavioral sensitivity is reflected by the antennal-specific increase in abundance of neural signaling transcripts in 14 days post-infection females, as determined by transcriptome analysis. This suggests that the sensitivity of the mosquito peripheral olfactory system is altered by the dengue virus by enhancing the overall neural responsiveness of the antenna, rather than the selective regulation of chemosensory-related genes. Our study reveals that dengue virus-1 enhances vector-related behaviors in the early stages post-infection that aid in avoiding predation and increasing spatial exploration. On the other hand, at the later stages of infection, the virus enhances the host-seeking capacity of the vector, thereby increasing the risk of virus transmission. A potential mechanism is discussed.

What Happens on Islands, doesn’t Stay on Islands: Patterns of Synchronicity in Mosquito Nuisance and Host-Seeking Activity between a Mangrove Island and Adjacent Coastal Development

Coastal development is expanding globally in response to mass human migration, yet urban planning guidelines often overlook the problems that human encroachment on or near coastal mosquito habitat may cause. This study aimed at elucidating the frequency and magnitude of dispersal of highly vagile saltmarsh mosquitoes from productive off-shore bay islands to adjacent coastal human developments. Inter-population dynamics and daily host-seeking activity of saltmarsh mosquitoes were monitored daily at 15-min intervals within a productive bay island and adjacent coastal development in southeast Queensland, Australia, using emerging smart trap technology over a 2-month period of high mosquito activity. The regulation of mosquito dispersal and host-seeking activity by local environmental factors, e.g. temperature, relative humidity and hourly wind patterns, were also investigated. The data show that the primary saltmarsh mosquitoes Aedes vigilax (Skuse) and Culex sitiens (Wiedemann) disperse from offshore breeding sites to neighboring mainland areas in high numbers and in highly synchronized waves despite unfavorable wind patterns and the need to traverse a considerable expanse (ca. 1.4 km) of open water. Patterns of host-seeking activity within each site were also remarkably similar despite notable differences in the local environment demonstrating a consistency in host-seeking activity across disparate habitats. These findings demonstrate that distant saltmarsh habitats, including offshore breeding sites, are likely to be primary sources of mosquito nuisance for coastal housing developments. This observation highlights the need to develop new planning and regulatory guidelines that alert urban planners to the risks of encroaching on habitats close to sources of highly vagile mosquito species.

Small-Molecule Agonists of Ae. aegypti Neuropeptide Y Receptor Block Mosquito Biting

Female Aedes aegypti mosquitoes bite humans to obtain blood to develop their eggs. Remarkably, their strong attraction to humans is suppressed for days after the blood meal by an unknown mechanism. We investigated a role for neuropeptide Y (NPY)-related signaling in long-term behavioral suppression and discovered that drugs targeting human NPY receptors modulate mosquito host-seeking. In a screen of all 49 predicted Ae. aegypti peptide receptors, we identified NPY-like receptor 7 (NPYLR7) as the sole target of these drugs. To obtain small-molecule agonists selective for NPYLR7, we performed a high-throughput cell-based assay of 265,211 compounds and isolated six highly selective NPYLR7 agonists that inhibit mosquito attraction to humans. NPYLR7 CRISPR-Cas9 null mutants are defective in behavioral suppression and resistant to these drugs. Finally, we show that these drugs can inhibit biting and blood-feeding on a live host, suggesting a novel approach to control infectious disease transmission by controlling mosquito behavior. VIDEO ABSTRACT.

Controlled release spatial repellent devices (CRDs) as novel tools against malaria transmission: a semi-field study in Macha, Zambia

BackgroundThe emergence of mosquitoes that can avoid indoor-deployed interventions, such as treated bed nets and indoor residual spraying, threatens the mainstay of malaria control in Zambia. Furthermore, the requirement for high coverage of these tools poses operational challenges. Spatial repellents are being assessed to supplement these vector control tools, but limitations exist in the residual effect of the repellent and the need for external power or heat for diffusion of the volatiles.MethodsA semi-field evaluation of a novel controlled release spatial repellent device (CRD) was conducted in Macha, Zambia. These devices emanate metofluthrin with no need for external power. Devices were deployed in huts within the semi-field system (SFS). Female Anopheles gambiae sensu stricto released within the SFS were trapped overnight by light traps and collected by aspiration the next morning inside and outside of huts to determine the extent of mosquito repellency and the impact on host-seeking and survival. Experiments studied the impact of number of devices as well as the presence of hut occupants. The study was complemented with numerical methods based on computational fluid dynamics to simulate spatial distribution of metofluthrin.ResultsPresence of CRDs was associated with significant reductions in indoor counts of mosquitoes, regardless of whether huts were occupied or not. Repellency ranged from 15 to 60% compared to huts with no devices. Reducing the number of devices from 16 to 4 had little impact on repellency. When huts were occupied, indoor mosquito host-seeking was higher in the presence of CRDs, whilst survival was significantly reduced.ConclusionsThis study demonstrated that deployment of as few as four CRDs within a hut was associated with reduced indoor mosquito densities. As would be expected, presence of occupants within huts, resulted in greater indoor catches (both with and without devices). The increased indoor mosquito host-seeking and mortality in huts when devices were present may be explained by the excito-repellency activity of metofluthrin. These semi-field experiments provide preliminary data on the utility of CRD spatial repellents to reduce indoor densities of An. gambiae mosquitoes. Studies will further investigate the impact of CRDs on mosquito behaviour as well as epidemiological protective efficacy.

An Efficient Adaptive Network-based Fuzzy Inference System with Mosquito Host-Seeking for Facial Expression Recognition

An Efficient Adaptive Network-based Fuzzy Inference System with Mosquito Host-Seeking for Facial Expression Recognition

Functional characterization of the dual allatostatin-A receptors in mosquitoes

The neuropeptide allatostatin-A (AstA) and its cognate receptors (AstARs) are involved in the modulation of feeding behavior, which in hematophagous insects includes the regulation of the disease vector-related behaviors, host seeking and blood feeding. In mosquitoes and other dipterans, there are two copies of AstAR, contrasting with the single copy found in other insects. In this study, we identified and cloned the dual AstAR system of two important disease vectors Aedes aegypti and Culex quinquefasciatus, and compared them with those previously described, including those in Anopheles coluzzii and Drosophila melanogaster. Phylogenetic analysis of the AstARs revealed that the mosquito AstAR1s has retained a similar amino acid sequence as the AstARs from non-dipteran insect species. Intron analysis revealed that the number of introns accumulated in the AstAR2s is similar to that in other insects, and that introns are conserved within the receptor types, but that only the final two introns are conserved across AstAR1s and 2s. We functionally characterized the dual AstARs in An. coluzzii, Ae. aegypti and Cx. quinquefasciatus by stably expressing the receptors in a Chinese hamster oocyte cell line (CHO) also stably expressing a promiscuous G-protein (G16), and challenged them with the endogenous isoforms of AstA from the three mosquito species. In the culicine mosquitoes, Ae. aegypti and Cx. quinquefasciatus, the AstARs demonstrated differential sensitivity to AstA, with the AstAR2s displaying a higher sensitivity than the AstAR1s, suggesting a divergence of functional roles for these AstARs. In contrast, both An. coluzzii AstARs demonstrated a similar sensitivity to the AstA ligands. We discuss our findings in the light of AstA acting as a regulator of blood feeding in mosquitoes. A better understanding of the regulation of host seeking and blood feeding in vector mosquitoes will lead to the rational development of novel approaches for vector control.