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.

Identification of candidate genes associated with host-seeking behavior in the parasitoid wasp Diachasmimorpha longicaudata.

BACKGROUND: Diachasmimorpha longicaudata is a hymenopteran fruit fly endoparasitoid. Females of this species find their hosts for oviposition by using complex sensorial mechanisms in response to physical and chemical stimuli associated with the host and host habitat. Ecological and behavioral aspects related to host-seeking behavior for oviposition have been extensively studied in D. longicaudata, including the identification of volatile organic compounds acting as attractants to females. In this sense, molecular mechanisms of chemoreception have been explored in this species, including a preliminary characterization of odorant-binding proteins (OBPs), chemosensory proteins (CSPs) and odorant receptors (ORs), among other proteins. Functional assays on OBP and CSP have been conducted as a first approach to identify molecular mechanisms associated with the female host-seeking behavior for oviposition. The aims of the present study were to identify the D. longicaudata sensory gene repertoire expressed in the antenna of sexually mature and mated individuals of both sexes, and subsequently, characterize transcripts differentially expressed in the antennae of females to identify candidate genes associated with the female host-seeking behavior for oviposition. RESULTS: A total of 33,745 predicted protein-coding sequences were obtained from a de novo antennal transcriptome assembly. Ten sensory-related gene families were annotated as follows: 222 ORs, 44 ionotropic receptors (IRs), 25 gustatory receptors (GRs), 9 CSPs, 13 OBPs, 2 ammonium transporters (AMTs), 8 pickpocket (PPKs) receptors, 16 transient receptor potential (TRP) channels, 12 CD36/SNMPs and 3 Niemann-Pick type C2 like proteins (NPC2-like). The differential expression analysis revealed 237 and 151 transcripts up- and downregulated, respectively, between the female and male antennae. Ninety-seven differentially expressed transcripts corresponded to sensory-related genes including 88 transcripts being upregulated (87 ORs and one TRP) and nine downregulated (six ORs, two CSPs and one OBP) in females compared to males. CONCLUSIONS: The sensory gene repertoire of D. longicaudata was similar to that of other taxonomically related parasitoid wasps. We identified a high number of ORs upregulated in the female antenna. These results may indicate that this gene family has a central role in the chemoreception of sexually mature females during the search for hosts and host habitats for reproductive purposes.

The Rpfor gene modulates the locomotory activity and host-seeking behaviour of Rhodnius prolixus.

The molecular bases of animal behaviour are intricate due to the pleiotropic nature of behaviour-modulating genes, which are often expressed across multiple tissues. The foraging gene (for) encodes a cGMP-dependent protein kinase (PKG), pivotal in regulating downstream target proteins through phosphorylation. In insects, for has been implicated in various behavioural contexts and physiological processes regarding searching for food. Rhodnius prolixus, a hematophagous bug that transmits Trypanosoma cruzi, the causative agent of Chagas disease, exhibits specific activity patterns associated with its hematophagous behaviour. Our previous work demonstrated a correlation between locomotor activity profiles and the expression of Rpfor, suggesting its involvement in modulating triatomine locomotion. In this study, we investigated the impact of Rpfor knockdown on locomotory activity, host-seeking behaviour, feeding performance and lipid metabolism in R. prolixus nymphs. Using RNA interference, we achieved a significant reduction of Rpfor expression in both the brain and fat body of R. prolixus nymphs. Knocked-down nymphs exhibited diminished non-oriented locomotory activity compared with controls, without altering the characteristic bimodal pattern of activity. Additionally, they displayed an increased tendency to approach a host, suggesting a role for Rpfor in modulating host-seeking behaviour. Feeding performance and lipid metabolism remained unaffected by Rpfor knockdown. Our findings underscore the multifaceted role of Rpfor in modulating locomotor activity and host-seeking behaviour in R. prolixus nymphs, shedding light on the molecular mechanisms underlying their hematophagous behaviour and potential implications for disease transmission. Further research is necessary to elucidate the intricate interplay between Rpfor expression, behaviour and physiological processes in triatomine bugs.

As bases moleculares do comportamento animal são complexas devido à natureza pleiotrópica dos genes envolvidos na sua modulação, normalmente expressos em múltiplos tecidos. O gene foraging (for) codifica para uma proteína quinase dependente de cGMP, fundamental para a regulação de proteínas alvo via fosforilação. Em insetos, o gene for tem sido associado a vários contextos comportamentais e processos fisiológicos relacionados com forrageamento. Rhodnius prolixus, um inseto hematófago que transmite Trypanosoma cruzi, o agente causativo da doença de Chagas, exibe padrões de atividade específicos associados com o seu comportamento hematófago. Em um estudo anterior, demonstramos uma correlação entre os perfis de atividade locomotora e a expressão de Rpfor, sugerindo o seu envolvimento na modulação da locomoção de triatomíneos. No presente estudo, investigamos o impacto do silenciamento de Rpfor na atividade locomotora, no comportamento de busca por hospedeiro, na performance alimentar, e no metabolismo de lipídeos em ninfas de R. prolixus. Através da técnica de RNA de interferência, obtivemos uma redução significativa da expressão do gene Rpfor no cérebro e no corpo gorduroso de R. prolixus. Insetos silenciados exibiram uma redução da atividade locomotora não orientada em comparação com controles, sem alterações no padrão bimodal da atividade. Adicionalmente, os insetos apresentaram um aumento no comportamento de busca por hospedeiro, sugerindo um papel para o Rpfor na sua modulação. A performance alimentar e o metabolismo de lipídeos não foram alterados pelo silenciamento do gene. Nossas descobertas ressaltam o papel multifuncional do gene Rpfor na modulação da atividade locomotora e no comportamento de busca por hospedeiro em R. prolixus, ampliando o conhecimento sobre os mecanismos moleculares relacionados ao seu comportamento hematófago e potenciais implicações para a transmissão de doenças. Estudos adicionais são necessários para elucidar a intrincada interação entre expressão, comportamento e processos fisiológicos de Rpfor em insetos triatomíneos.

The black-legged tick Ixodes scapularis detects CO2 without the Haller's organ.

Both male and female ticks have a strong innate drive to find and blood-feed on hosts. Carbon dioxide (CO2) is considered a critical behavioral activator and attractant for ticks and an essential sensory cue to find hosts. Yet, how CO2 activates and promotes host seeking in ticks is poorly understood. CO2 responses were studied in the black-legged tick Ixodes scapularis, the primary vector for Lyme disease in North America. Adult males and females were exposed to 1%, 2%, 4% or 8% CO2, and changes in walking behavior and foreleg movement were analyzed. CO2 is a potent stimulant for adult I. scapularis, even at lower concentrations (1%). Behavioral reactions depended on the animal's state: walking ticks increased their walking speed, while stationary ticks started to wave their forelegs and began to quest - both behaviors resembling aspects of host seeking. Only in sporadic cases did stationary animals start to walk when exposed to CO2, supporting the hypothesis that CO2 acts as an activator rather than an attractant. Furthermore, I. scapularis did not show a clear concentration preference and was not tuned more robustly to breath-like CO2 concentrations (∼4%) than to the other concentrations tested. Moreover, convincing evidence is provided showing that the foreleg Haller's organ is not necessary for CO2 detection. Even with a disabled or amputated Haller's organ, I. scapularis responded robustly to CO2, signifying that there must be CO2-sensitive structures important for tick host seeking that have not yet been identified.

Soil moisture conditions alter behavior of entomopathogenic nematodes.

BACKGROUND: A variety of environmental factors can disrupt biotic interactions between plants, insects and soil microorganisms with consequences for agricultural management and production. Many of these belowground interactions are mediated by volatile organic compounds (VOCs) which can be used for communication under appropriate environmental conditions. Behavioral responses to these compounds may likewise be dependent on varying soil conditions which are influenced by a changing climate. To determine how changing environmental conditions may affect VOC-mediated biotic interactions, we used a belowground system where entomopathogenic nematodes (EPNs) - tiny roundworm parasitoids of soil-borne insects - respond to VOCs by moving through the soil pore matrix. Specifically, we used two genera of EPNs - Heterorhabditis and Steinernema - that are known to respond to four specific terpenes - α-pinene, linalool, d-limonene and pregeijerene - released by the roots of plants in the presence of herbivores. We assessed the response of these nematodes to these terpenes under three moisture regimes to determine whether drier conditions or inundated conditions may influence the response behavior of these nematodes. RESULTS: Our results illustrate that the recovery rate of EPNs is positively associated with soil moisture concentration. As soil moisture concentration increases from 6% to 18%, substantially more nematodes are recovered from bioassays. In addition, we find that soil moisture influences EPN preference for VOCs, as illustrated in the variable response rates. Certain compounds shifted from acting as a repellent to acting as an attractant and vice versa depending on the soil moisture concentration. CONCLUSION: On a broad scale, we demonstrate that soil moisture has a significant effect on EPN host-seeking behavior. EPN efficacy as biological control agents could be affected by climate change projections that predict varying soil moisture concentrations. We recommend that maintaining nematodes as biological control agents is essential for sustainable agriculture development, as they significantly contribute not only to soil health but also to efficient pest management. © 2024 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Why are biting flies attracted to blue objects?

Diurnal biting flies are strongly attracted to blue objects. This behaviour is widely exploited for fly control, but its functional significance is debated. It is hypothesized that blue objects resemble animal hosts; blue surfaces resemble shaded resting places; and blue attraction is a by-product of attraction to polarized light. We computed the fly photoreceptor signals elicited by a large sample of leaf and animal integument reflectance spectra, viewed under open/cloudy illumination and under woodland shade. We then trained artificial neural networks (ANNs) to distinguish animals from leaf backgrounds, and shaded from unshaded surfaces, in order to find the optimal means of doing so based upon the sensory information available to a fly. After training, we challenged ANNs to classify blue objects used in fly control. Trained ANNs could make both discriminations with high accuracy. They discriminated animals from leaves based upon blue-green photoreceptor opponency and commonly misclassified blue objects as animals. Meanwhile, they discriminated shaded from unshaded stimuli using achromatic cues and never misclassified blue objects as shaded. We conclude that blue-green opponency is the most effective means of discriminating animals from leaf backgrounds using a fly's sensory information, and that blue objects resemble animal hosts through such mechanisms.

Anatomical, phenological and genetic aspects of the host-parasite relationship between Andrena vaga (Hymenoptera) and Stylops ater (Strepsiptera).

Stylops ater is an endoparasite of the mining bee Andrena vaga with extreme sexual dimorphism and hypermetamorphosis. Its population structure, parasitization mode, genetic diversity and impact on host morphology were examined in nesting sites in Germany to better understand this highly specialized host­parasite interaction. The shift in host emergence due to stylopization was proven to be especially strong in A. vaga. Around 10% of bees hosted more than 1 Stylops, with at maximum 4. A trend in Stylops' preference for hosts of their own sex and a sex-specific position of extrusion from the host abdomen was found. Invasion of Andrena eggs by Stylops primary larvae was depicted for the first time. Cephalothoraces of female Stylops were smaller in male and pluristylopized hosts, likely due to lower nutrient supply. The genes H3, 18S and cytochrome c oxidase subunit 1 were highly conserved, revealing near-absence of local variation within Stylops. Ovaries of hosts with male Stylops contained poorly developed eggs while those of hosts with female Stylops were devoid of visible eggs, which might be due to a higher protein demand of female Stylops. Male Stylops, which might have a more energy-consuming development, led to a reduction in head width of their hosts. Host masculinization was present in the leaner shape of the metabasitarsus of stylopized females and is interpreted as a by-product of manipulation of the host's endocrine system to shift its emergence. Stylopization intensified tergal hairiness, most strongly in hosts with female Stylops, near the point of parasite extrusion, hinting towards substance-induced host manipulation.

Chemosensory mechanisms of host seeking and infectivity in skin-penetrating nematodes.

Approximately 800 million people worldwide are infected with one or more species of skin-penetrating nematodes. These parasites persist in the environment as developmentally arrested third-stage infective larvae (iL3s) that navigate toward host-emitted cues, contact host skin, and penetrate the skin. iL3s then reinitiate development inside the host in response to sensory cues, a process called activation. Here, we investigate how chemosensation drives host seeking and activation in skin-penetrating nematodes. We show that the olfactory preferences of iL3s are categorically different from those of free-living adults, which may restrict host seeking to iL3s. The human-parasitic threadworm Strongyloides stercoralis and hookworm Ancylostoma ceylanicum have highly dissimilar olfactory preferences, suggesting that these two species may use distinct strategies to target humans. CRISPR/Cas9-mediated mutagenesis of the S. stercoralis tax-4 gene abolishes iL3 attraction to a host-emitted odorant and prevents activation. Our results suggest an important role for chemosensation in iL3 host seeking and infectivity and provide insight into the molecular mechanisms that underlie these processes.

The antennal transcriptome of Triatoma infestans reveals substantial expression changes triggered by a blood meal.

BACKGROUND: Triatoma infestans is the main vector of Chagas disease in the Americas, currently transmitting it in Argentina, Paraguay, and Bolivia. Many T. infestans populations present insecticide resistance, reducing the efficiency of control campaigns. Alternative vector control methods are needed, and molecular targets mediating fundamental physiological processes can be a promising option to manipulate kissing bug behavior. Therefore, it is necessary to characterize the main sensory targets, as well as to determine whether they are modulated by physiological factors. In order to identify gene candidates potentially mediating host cue detection, the antennal transcripts of T. infestans fifth instar larvae were sequenced and assembled. Besides, we evaluated whether a blood meal had an effect on transcriptional profiles, as responsiveness to host-emitted sensory cues depends on bug starvation. RESULTS: The sensory-related gene families of T. infestans were annotated (127 odorant receptors, 38 ionotropic receptors, 11 gustatory receptors, 41 odorant binding proteins, and 25 chemosensory proteins, among others) and compared to those of several other hemipterans, including four triatomine species. Several triatomine-specific lineages representing sensory adaptations developed through the evolution of these blood-feeding heteropterans were identified. As well, we report here various conserved sensory gene orthogroups shared by heteropterans. The absence of the thermosensor pyrexia, of pickpocket receptor subfamilies IV and VII, together with clearly expanded takeout repertoires, are revealed features of the molecular bases of heteropteran antennal physiology. Finally, out of 2,122 genes whose antennal expression was significantly altered by the ingestion of a blood meal, a set of 41 T. infestans sensory-related genes (9 up-regulated; 32 down-regulated) was detected. CONCLUSIONS: We propose that the set of genes presenting nutritionally-triggered modulation on their expression represent candidates to mediate triatomine host-seeking behavior. Besides, the triatomine-specific gene lineages found represent molecular adaptations to their risky natural history that involves stealing blood from an enormously diverse set of vertebrates. Heteropteran gene orthogroups identified may represent unknown features of the sensory specificities of this largest group of hemipteroids. Our work is the first molecular characterization of the peripheral modulation of sensory processes in a non-dipteran vector of human disease.

Chemotactic and temperature-dependent responses of the Strongyloidoidea superfamily of nematodes.

Host-seeking behaviour and how a parasite identifies the correct host to infect remains a poorly understood area of parasitology. What is currently known is that host sensation and seeking behaviour is formed from a complex mixture of chemo-, thermo- and mechanosensory behaviours, of which chemosensation is the best studied. Previous studies of olfaction in parasitic nematodes suggested that this behaviour appears to be more closely related to target host and infection mode than phylogeny. However, there has not yet been a study comparing the chemotactic and temperature-dependent behaviours of very closely related parasitic and non-parasitic nematodes. To this end, we examined the temperature-dependent and chemotactic responses of the Strongyloidoidea superfamily of nematodes. We found differences in temperature response between the different species and within infective larvae. Chemotactic responses were highly divergent, with different attraction profiles between all species studied. When examining direct stimulation with fur, we found that it was insufficient to cause an attractive response. Overall, our results support the notion that olfactory sensation is more closely related to lifestyle and host range than phylogeny, and that multiple cues are required to initiate host-seeking behaviour.

Salivary AsHPX12 influences pre-blood meal associated behavioral properties in Anopheles stephensi.

BACKGROUND & OBJECTIVES: A successful blood meal acquisition process by an adult female mosquito is accomplished through salivary glands, which releases a cocktail of proteins to counteract the vertebrate host's immune homeostasis. Here, we characterize a salivary-specific Heme peroxidase family member HPX12, originally identified from Plasmodium vivax infected salivary RNAseq data of the mosquito Anopheles stephensi. METHODS: To demonstrate we utilized a comprehensive in silico and functional genomics approach. RESULTS: Our dsRNA-mediated silencing experiments demonstrate that salivary AsHPX12 may regulate pre-blood meal-associated behavioral properties such as probing time, probing propensity, and host attraction. Altered expression of the salivary secretory and antennal proteins expression may have accounted for salivary homeostasis disruption resulting in the unusual fast release of salivary cocktail proteins and delayed acquisition of blood meal in the AsHPX12 knockdown mosquitoes. We also observed a significant parallel transcriptional modulation in response to blood feeding and P. vivax infection. INTERPRETATION & CONCLUSION: With this work, we establish a possible functional correlation of AsHPX12 role in the maintenance of salivary physiological-homeostasis, and Plasmodium sporozoites survival/transmission, though the mechanism is yet to unravel.

Modulation of odour-guided behaviour in mosquitoes.

Mosquitoes are emerging as model systems with which to study innate behaviours through neuroethology and functional genomics. Decades of work on these disease vectors have provided a solid behavioural framework describing the distinct repertoire of predominantly odour-mediated behaviours of female mosquitoes, and their dependence on life stage (intrinsic factors) and environmental cues (extrinsic factors). The purpose of this review is to provide an overview of how intrinsic factors, including adult maturation, age, nutritional status, and infection, affect the attraction to plants and feeding on plant fluids, host seeking, blood feeding, supplemental feeding behaviours, pre-oviposition behaviour, and oviposition in female mosquitoes. With the technological advancements in the recent two decades, we have gained a better understanding of which volatile organic compounds are used by mosquitoes to recognise and discriminate among various fitness-enhancing resources, and characterised their neural and molecular correlates. In this review, we present the state of the art of the peripheral olfactory system as described by the neural physiology, functional genomics, and genetics underlying the demonstrated changes in the behavioural repertoire in female mosquitoes. The review is meant as a summary introduction to the current conceptual thinking in the field.

Olfaction in Anopheles mosquitoes.

As vectors of disease, mosquitoes are a global threat to human health. The Anopheles mosquito is the deadliest mosquito species as the insect vector of the malaria-causing parasite, which kills hundreds of thousands every year. These mosquitoes are reliant on their sense of smell (olfaction) to guide most of their behaviors, and a better understanding of Anopheles olfaction identifies opportunities for reducing the spread of malaria. This review takes a detailed look at Anopheles olfaction. We explore a range of topics from chemosensory receptors, olfactory neurons, and sensory appendages to behaviors guided by olfaction (including host-seeking, foraging, oviposition, and mating), to vector management strategies that target mosquito olfaction. We identify many research areas that remain to be addressed.

Entomological survey in two communes with residual malaria transmission in Gia Lai Province in the central highlands of Vietnam.

BACKGROUND: In 2018, the National Malaria Control Programme in Vietnam switched from prioritizing malaria control to elimination. However, with the ongoing elimination programme, there are still areas where residual malaria transmission persists, including the central highlands. This entomological survey was conducted to evaluate Anopheles diversity and host-seeking activity of Anopheles vectors in two communes with very low malaria transmission in Gia Lai Province. METHODS: Anopheles species were collected in Ia DReh commune and Ia KDam commune, Gia Lai Province in the central highlands of Vietnam. Collections were conducted using human-baited double net trap, light trap and manual aspiration collections around cattle sheds, in the dry and rainy season. Mosquito specimens were identified morphologically, and members of species complexes were distinguished molecularly. Mosquito night-feeding patterns were investigated during the dry and rainy seasons. RESULTS: Overall, 18,835 specimens including 19 taxa were collected in Ia KDam and Ia DReh communes. These included the primary malaria vectors, Anopheles dirus and Anopheles minimus, and other secondary vector species. Anopheles dirus was observed to be an anthropophilic species, whereas An. minimus and a number of secondary vectors were observed to be zoophilic. Anopheles vagus was the dominant species, followed by Anopheles sinensis and Anopheles peditaeniatus. The majority of specimens were collected in the rainy season due to the relatively large number of An. vagus, while An. peditaeniatus, An. dirus, Anopheles kochi, Anopheles monstrosus and Anopheles tessellatus were collected in greater numbers during the dry season. The peak of host-seeking activity for An. dirus, An. sinensis, and An. vagus was between 18.00 and 19.00 h. CONCLUSION: This study provided information on the diversity, seasonal prevalence and behaviour of Anopheles at the study sites. Identifying the diverse mosquito fauna in the central highlands of Vietnam allows species-specific control measures to be implemented by the National Programme to reduce malaria in areas of very low malaria transmission. The peak Anopheles host-seeking activity observed in this study was between 18.00 and 23.00 h, which highlights the need to better characterize Anopheles behaviour in this region of Vietnam to inform on vector control strategies.

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

BACKGROUND: Advances 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. METHODS: Quantified 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. RESULTS: In 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. CONCLUSIONS: Results 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.

Efficiency of the CO2 -baited omni-directional Fay-Prince trap under semi-field conditions and characterizing response behaviours for the yellow fever mosquito, Aedes aegypti.

Aedes aegypti (L.) (Diptera: Culicidae) is a vector of several serious disease-causing viruses including Dengue, Zika, chikungunya and yellow fever. Effective and efficient trapping methods are essential for meaningful mosquito population and disease-presence surveillance and ultimately, vector control. The Fay-Prince trap (FPT) was developed in the late 1960s as a daytime visual trap for male Ae. aegypti. Since then, its use has been expanded into the trapping of female Ae. aegypti, Aedes albopictus Skuse, other Ae. spp., and Culex spp. The efficiency of the FPT alone and with CO2 was tested under semi-field conditions and the behaviour of responding female Ae. aegypti was characterized. The mean capture efficiency of the FPT with CO2 per 30 min in the greenhouse was 3.07% and the capture rate from the total number of mosquitoes in our semi-field setup was slightly higher at 4.45%. Understanding the behaviours that mosquitoes exhibit during their encounter with particularly a visual trap may recommend trap improvements and contributes to our understanding of host-seeking behaviour and how it might be exploited.

Evaluation of human-baited double net trap and human-odour-baited CDC light trap for outdoor host-seeking malaria vector surveillance in Kenya and Ethiopia.

BACKGROUND: Surveillance of outdoor host-seeking malaria vectors is crucial to monitor changes in vector biting behaviour and evaluate the impact of vector control interventions. Human landing catch (HLC) has been considered the most reliable and gold standard surveillance method to estimate human-biting rates. However, it is labour-intensive, and its use is facing an increasing ethical concern due to potential risk of exposure to infectious mosquito bites. Thus, alternative methods are required. This study was conducted to evaluate the performance of human-odour-baited CDC light trap (HBLT) and human-baited double net trap (HDNT) for outdoor host-seeking malaria vector surveillance in Kenya and Ethiopia. METHODS: The sampling efficiency of HBLT and HDNT was compared with CDC light trap and HLC using Latin Square Design in Ahero and Iguhu sites, western Kenya and Bulbul site, southwestern Ethiopia between November 2015 and December 2018. The differences in Anopheles mosquito density among the trapping methods were compared using generalized linear model. RESULTS: Overall, 16,963 female Anopheles mosquitoes comprising Anopheles gambiae sensu lato (s.l.), Anopheles funestus s.l., Anopheles pharoensis, Anopheles coustani and Anopheles squamosus were collected. PCR results (n = 552) showed that Anopheles arabiensis was the only member of An. gambiae s.l. in Ahero and Bulbul, while 15.7% An. arabiensis and 84.3% An. gambiae sensu stricto (s.s.) constituted An. gambiae s.l. in Iguhu. In Ahero, HBLT captured 2.23 times as many An. arabiensis and 2.11 times as many An. funestus as CDC light trap. In the same site, HDNT yielded 3.43 times more An. arabiensis and 3.24 times more An. funestus than HBLT. In Iguhu, the density of Anopheles mosquitoes did not vary between the traps (p > 0.05). In Bulbul, HBLT caught 2.19 times as many An. arabiensis as CDC light trap, while HDNT caught 6.53 times as many An. arabiensis as CDC light trap. The mean density of An. arabiensis did not vary between HDNT and HLC (p = 0.098), whereas the HLC yielded significantly higher density of An. arabiensis compared to HBLT and CDC light trap. There was a significant density-independent positive correlation between HDNT and HLC (r = 0.69). CONCLUSION: This study revealed that both HBLT and HDNT caught higher density of malaria vectors than conventional CDC light trap. Moreover, HDNT yielded a similar vector density as HLC, suggesting that it could be an alternative tool to HLC for outdoor host-seeking malaria vector surveillance.

A comparative evaluation of northern and southern Ixodes scapularis questing height and hiding behaviour in the USA.

Ticks display a distinct type of host-seeking behaviour called questing. It has been proposed that the questing behaviour of Ixodes scapularis explains the geographic variation in Lyme disease (LD) risk in the eastern USA because the northern population has been shown to quest more often than the southern population. The height at which questing occurs is variable and this study aimed to characterize questing height for I. scapularis. Ticks were collected from a northern and southern state (i.e. Maryland and Texas) and bioassays were conducted. We report that nymphs from Texas quested at lower heights compared to nymphs from Maryland. In addition, only Texas nymphs exhibited a behaviour we call 'hiding behaviour'. These results may reflect the different composition of hosts between these two areas as the south has a higher abundance of lizards. In contrast, there was no significant difference in questing height between Maryland adults and Texas adults which was to be expected since adults are feeding on white-tailed deer in both locations. If all southern I. scapularis nymphs are questing at lower heights, this might make them less likely to come into contact with humans and this may be contributing to the geographical difference in LD prevalence.

Aversion of the invasive Asian longhorned tick to the white-footed mouse, the dominant reservoir of tick-borne pathogens in the U.S.A.

The Asian longhorned tick (Haemaphysalis longicornis) was reported for the first time in the U.S.A. in 2017 and has now spread across 12 states. The potential of this invasive tick vector to transmit pathogens will be determined through its association to hosts, such as the white-footed mouse (Peromyscus leucopus), which is the primary reservoir for the causative agent of Lyme disease (Borrelia burgdorferi) and other zoonotic pathogens. Larval H. longicornis were placed on P. leucopus; 65% of the larvae (n = 40) moved off the host within a short period of time, and none engorged. By contrast, larval blacklegged ticks (Ixodes scapularis) did not move from where they were placed in the ear of the mouse. A laboratory behavioural assay was then conducted to assess the interaction of H. longicornis with the hair of potential mammalian host species in the U.S.A. H. longicornis larvae were significantly less likely to enter the hair zone of P. leucopus and humans compared to the hair of domestic cats, domestic dogs and white-tailed deer. This study identifies a tick-host interaction behaviour, which can be quantified in a laboratory assay to predict tick-host associations and provides insights into how ticks select a host.

Early Questing by Lone Star Tick Larvae, New York and Massachusetts, USA, 2018.

Subtropical lone star tick larvae typically emerge in late summer. We found clusters of host-seeking lone star tick larvae during early June 2018 in New York and Massachusetts, USA. Invasion and persistence of this tick in more northern locations may have been promoted by adaptation to an accelerated life cycle.