Mosquito Response Research Articles

Transcriptomic analysis and molecular docking reveal genes involved in the response of Aedes aegypti larvae to an essential oil extracted from Eucalyptus.

BackgroundAedes aegypti (L.) is an urban mosquito, vector of several arboviruses that cause severe diseases in hundreds of million people each year. The resistance to synthetic insecticides developed by Ae. aegypti populations worldwide has contributed to failures in vector control campaigns, increasing the impact of arbovirus diseases. In this context, plant-derived essential oils with larvicidal activity could be an attractive alternative for vector control. However, the mode of action and the detoxificant response of mosquitoes to plant derived compounds have not been established, impairing the optimization of their use.Methods and findingsHere we compare gene expression in Ae. aegypti larvae after 14 hrs of exposure to Eucalyptus camaldulensis essential oil with a control group exposed to vehicle (acetone) for the same lapse, by using RNA-Seq. We found differentially expressed genes encoding for cuticle proteins, fatty-acid synthesis, membrane transporters and detoxificant related gene families (i.e. heat shock proteins, cytochromes P450, glutathione transferases, UDP-glycosyltransferases and ABC transporters). Finally, our RNA-Seq and molecular docking results provide evidence pointing to a central involvement of chemosensory proteins in the detoxificant response in mosquitoes.Conclusions and significanceOur work contributes to the understanding of the physiological response of Ae. aegypti larvae to an intoxication with a natural toxic distilled from Eucalyptus leafs. The results suggest an involvement of most of the gene families associated to detoxification of xenobiotics in insects. Noteworthy, this work provides important information regarding the implication of chemosensory proteins in the detoxification of a natural larvicide. Understanding the mode of detoxification of Eucalyptus distilled compounds could contribute to their implementation as a tool in mosquito control.

Geographic variation in Culex oviposition habitat selection responses to a predator, Notonecta irrorata

1. Predators have effects on prey populations through both consumptive and non‐consumptive effects. Predator's presence is expected to drive variation in prey oviposition habitat selection behaviour, but differences in biotic and abiotic characteristics of habitats, or trait variation, may produce geographic variation in species interactions.2. We conducted a series of experiments in two geographic locations, Mississippi and Missouri, USA, to assess oviposition responses of Culex mosquitoes (prey) to the presence of Notonecta irrorata (predator). We first tested whether mosquitoes in each location respond to the presence of N. irrorata, with follow‐up experiments to determine whether mosquitoes respond to variation in N. irrorata density, whether N. irrorata from each location generate different responses by the same Culex population, and whether diet and consumption of conspecifics affect oviposition.3. We found that Culex restuans in Missouri had reduced oviposition when N. irrorata were present. In Mississippi, C. restuans did not respond to the presence of N. irrorata from either Mississippi or Missouri, to the variation in density of N. irrorata, or to N. irrorata that had been fed larval mosquitoes.4. Our study documents the first instance of geographic variation in oviposition response of a prey species to a predator species.

Effect of Different Wall Surface Coverage With Deltamethrin-Treated Netting on the Reduction of Indoor-Biting Anopheles Mosquitoes (Diptera: Culicidae).

Indoor residual spray with deltamethrin remains the most common tool for reducing malaria transmission in Thailand. Deltamethrin is commonly used to spray the entire inner surfaces of the walls to prevent mosquitoes from resting. This study compared the mosquito landing responses on humans inside three experimental huts treated with deltamethrin at three different extents of wall coverage (25%, 50%, and full coverage), with one clean/untreated hut serving as a control. There were no significant differences between the numbers of Anopheles mosquitoes landing in the 50% and full coverage huts, whereas, in comparison to both of these, there was a significantly greater number landing in the 25% coverage hut. This study demonstrates that varying the percent coverage of indoor surfaces with deltamethrin-treated netting influences the blood-feeding success of wild Anopheles, and our findings suggest that it may be possible to reduce the extent of insecticide surface treatment while maintaining equivalent mosquito avoidance action to that seen in fully treated structures.

Evaluation of Mosquito Attractant Candidates Using a High-Throughput Screening System for Aedes aegypti (L.), Culex quinquefasciatus Say. and Anopheles minimus Theobald (Diptera: Culicidae).

Simple SummaryTrapping mosquitoes can enhance its capture rate by adding attractants such as carbon dioxide or human hosts’ odor-mimicking synthetic blends. Various olfactometers exist to test mosquitos’ behavior, but high-throughput screening system (HITSS)—one of the diffusion assays—has not been applied to developing lures. In this study, six different newly prepared chemical lure candidates (Kasetsart University (KU)-lures) were tested for diurnal Aedes aegypti, nocturnal Culex quinquefasciatus and nocturnal Anopheles minimus, using the HITSS assay. Results showed species-specific different lure preferences; the diurnal species were attracted to KU-lure #1 (29.7%), while both of the nocturnal species preferred KU-lure #6 (68.3% and 74.3% for Cx. quinquefasciatus and An. minimus, respectively). In addition, the selected lure candidates clearly demonstrated dose-dependent reversal responses for each Ae. aegypti and Cx. quinquefasciatus. Our results indicate that the HITSS assay distinguishes potential species-specific lure candidates. In addition, the HITSS assay was equally effective in determining the host-seeking behavior in pyrethroid-resistant and -susceptible strains. Further studies are needed to determine the accuracy of the HITSS assay in large-scale semi-field screen house tests using commercial traps.Several types of olfactometers have been used to evaluate mosquito responses to agents that mimic natural volatiles that repel or attract. The Y-tube olfactometer has been widely used to study repellents and attractants, while the high-throughput screening system assay has only been used to study repellents. Whether the high-throughput screening system assay is suitable for evaluating attractants is unknown. We evaluated the responses to four lactic-acid-based mixtures and two non-lactic-acid-based chemical lure candidates using the high-throughput screening system (HITSS) for three mosquito species (laboratory strains and field populations of both Aedes aegypti (L.) and Culex quinquefasciatus Say.; laboratory strain of Anopheles minimus Theobald) under laboratory-controlled conditions. HITSS assay results showed that KU-lure #1 elicited the greatest percent attraction for pyrethroid-resistant and -susceptible Ae. aegypti. KU-lure #6 elicited the strongest attractive response for pyrethroid-susceptible and -resistant Cx. quinquefasciatus and pyrethroid-susceptible An. minimus. The response to the lures from each species was independent of the pyrethroid susceptibility status (Ae. aegypti, p = 0.825; Cx. quinquefasciatus, p = 0.056). However, a significant difference in attraction to KU-lure #6 was observed between diurnal and nocturnal mosquitoes (Cx. quinquefasciatus vs. Ae. aegypti, p = 0.014; An. minimus vs. Ae. aegypti, p = 0.001). The laboratory-level HITSS assay effectively selects potential lure candidates. Because the host-seeking behavior differs between mosquito species, further studies are needed to develop species-specific attractants. Additional studies in semi-field screen houses using commercial traps are necessary to evaluate the accuracy of these laboratory assay results.

Response of Mosquitoes Associated with Estuarine Wetlands to Bushfire in Australia.

The response of mosquitoes to bushfire is poorly understood. During the 2019-20 summer, many regions of Australia were impacted by devastating bushfires. An area of estuarine and brackish-water wetlands alongside the Georges River, Sydney, New South Wales, was burned in January 2020. Mosquito populations within the area were monitored as part of the local authority's mosquito management program, providing a unique opportunity to record the response of key mosquitoes of pest and public health concern to bushfire. Ground pools within a tidally influenced swamp oak forest dominated by Casuarina glauca and associated wetlands dominated by Phragmites australis and Bolboschoenus spp. had been identified as suitable habitat for a range of mosquitoes, including Aedes alternans, Ae. vigilax, and Verrallina funerea. Surveys of immature stages of mosquitoes within recently burned habitats inundated by tides demonstrated that mosquito eggs survived the direct and indirect impacts of fire and immature stages successfully completed development as reflected in concomitant changes in adult mosquito populations following the bushfire. This unique observation has implications for mosquito management following bushfire in Australia and internationally.

Distinct Roles of Hemocytes at Different Stages of Infection by Dengue and Zika Viruses in Aedes aegypti Mosquitoes.

Aedes aegypti mosquitoes are vectors for arboviruses of medical importance such as dengue (DENV) and Zika (ZIKV) viruses. Different innate immune pathways contribute to the control of arboviruses in the mosquito vector including RNA interference, Toll and Jak-STAT pathways. However, the role of cellular responses mediated by circulating macrophage-like cells known as hemocytes remains unclear. Here we show that hemocytes are recruited to the midgut of Ae. aegypti mosquitoes in response to DENV or ZIKV. Blockade of the phagocytic function of hemocytes using latex beads induced increased accumulation of hemocytes in the midgut and a reduction in virus infection levels in this organ. In contrast, inhibition of phagocytosis by hemocytes led to increased systemic dissemination and replication of DENV and ZIKV. Hence, our work reveals a dual role for hemocytes in Ae. aegypti mosquitoes, whereby phagocytosis is not required to control viral infection in the midgut but is essential to restrict systemic dissemination. Further understanding of the mechanism behind this duality could help the design of vector-based strategies to prevent transmission of arboviruses.

The Effect of the Acoustic Swarm Environment on the Behaviour and Hearing of Male Culex quinquefasciatus Mosquitoes

Background The mating behaviour of many mosquito species is mediated by sound: males pursue and mate with a female mid-flight by using their antennae to detect and track the whine of the female flight-tone. Upon detection of the female's flight-tones (or artificial female-like tones), males track the sound source and display stereotypical rapid frequency modulation (RFM) acoustic behaviour when very close to the female. Under natural conditions, flying males of these mosquito species form relatively dense agglomerations – swarms – while waiting for sexually receptive females to pass nearby. Individual male mosquitoes thus face intense competition to detect, track, and access a passing female. Aims We aim to understand the significance of the swarm acoustics on the individual mating behaviour of male mosquitoes. Specifically, we tested the influence of simulated swarm sound on the RFM behaviour and on the hearing physiology of male mosquitoes. Methods We derived the behavioural threshold responses of individual male mosquitoes exposed to female-like tones with and without the presence of a banded 700-900 Hz synthesised swarm sound. Two electrophysiological experiments were also performed: 1) measurement of the threshold of the mosquito auditory organ, the Johnston's organ (JO), to female-like tones with and without swarm sound; 2) measurement from the JO of electrical difference-tone distortion products generated by tones at the frequencies of the male and female flight tones. Results Behavioural experiments with free-flying mosquitoes and electrophysiological preparations indicate that swarm sound has no apparent affect on the behavioural and hearing thresholds of males for female-like sounds. Conclusion This indicates that from an individual male mosquito perspective, swarms are not a source of acoustic noise and males are neither facilitated nor impaired by synthetic swarm noise in detecting females. Furthermore, the swarm noise does not appear to impair or facilitate the ability of males to detect their own flight tones and those of nearby male mosquitoes.

Multimodal synergisms in host stimuli drive landing response in malaria mosquitoes

Anopheles mosquitoes transmit malaria, which affects one-fifth of the world population. A comprehensive understanding of mosquito behaviour is essential for the development of novel tools for vector control and surveillance. Despite abundant research on mosquito behaviour, little is known on the stimuli that drive malaria vectors during the landing phase of host-seeking. Using behavioural assays with a multimodal step approach we quantified both the individual and the combined effect of three host-associated stimuli in eliciting landing in Anopheles coluzzii females. We demonstrated that visual, olfactory and thermal sensory stimuli interact synergistically to increase the landing response. Furthermore, if considering only the final outcome (i.e. landing response), our insect model can bypass the absence of either a thermal or a visual stimulus, provided that at least one of these is presented simultaneously with the olfactory stimuli, suggesting that landing is the result of a flexible but accurate stimuli integration. These results have important implications for the development of mosquito control and surveillance tools.

Influence of Vertebrate Excreta on Attraction, Oviposition and Development of the Asian Tiger Mosquito, Aedes albopictus (Diptera: Culicidae)

Simple SummaryCommonly known as the Asian tiger mosquito, Aedes albopictus is a vector of dengue worldwide. Knowledge of the behavior of dengue vectors facilitates effective vector control. This is the first comprehensive analysis of selected vertebrate excreta of goat, cow and pig to identify the oviposition attraction and growth performance of Ae. albopictus in Sri Lanka. The current study revealed that Ae. albopictus gravid females are significantly attracted to goat excreta but are repelled by pig excreta. The oviposition preference was highest for the cow excreta and lowest for the pig excreta. For excreta combinations, the Cow+Goat combination increased the oviposition while the Pig+Goat combination reduced the oviposition. The oviposition preference of Ae. albopictus increased with the rate of fermentation. The pig excreta increased the Ae. albopictus larval mortality, larval and pupal duration and reduced adult fecundity, whereas the cow excreta positively affected all these aspects. Additionally, our findings suggest that a high abundance of Ae. albopictus in rural areas of Sri Lanka is possibly due to its oviposition attraction and the growth performance of the vertebrate excreta.Aedes albopictus is an important vector of dengue worldwide. Eliminating dengue in Sri Lanka depends entirely on controlling the vector and human-vector contact. Thus, studying the bionomics and behavior of Ae. albopictus is paramount. The objective of this study was to evaluate the effect of the excreta of cow, goat and pig on the attraction, oviposition and development of Ae. albopictus. Bioassay chambers determined the mosquito stimulatory response. Ovitraps determined Ae. albopictus oviposition preference to excreta singly, in combination and on fermentation. The excreta effect on larval development was also determined. The results revealed that Ae. albopictus gravid females were significantly attracted to goat excreta but were repelled by pig excreta. The oviposition preference was highest for cow excreta and lowest for pig excreta. For excreta combinations, the Cow+Goat combination increased the oviposition while the Pig+Goat combination reduced the oviposition. The oviposition preference of Ae. albopictus increased with the rate of fermentation. The pig excreta increased the Ae. albopictus larval mortality, larval and pupal duration and reduced adult fecundity, whereas the cow excreta positively affected all these aspects. Our findings additionally suggest that a high abundance of Ae. albopictus in rural areas of Sri Lanka may be due to its oviposition attraction and growth performance for vertebrate excreta.

Mosquito responses to lethal and nonlethal effects of predation and an insect growth regulator

AbstractLethal and nonlethal effects of predation are likely to impact phenotypic traits of potential prey that elude capture. Nonlethal effects of predation on prey can include alterations in behavior or morphology in response to predation risk which may impair prey growth and fitness. The application of pesticide in mosquito control during aquatic stages can interact with predation stress and alter density and phenotypic traits of prey. Insect growth regulator (IGR) pyriproxyfen is a pesticide that mainly prevents pupal‐adult metamorphosis by mimicking juvenile hormone, whereas the larval stage is not targeted. The use of IGR can therefore act in conjunction with natural aquatic predators that target the larval stage to affect population of prey. In this study, we assessed the invasive mosquito prey Aedes aegypti responses to lethal and nonlethal effects of a combination of IGR and predatory mosquito larvae of Toxorhynchites rutilus. The combination of IGR and Tx. rutilus heavily lowered Ae. aegypti metamorphosis to adulthood more than the independent effects of IGR or Tx. rutilus. Exposing Ae. aegypti larvae to the combination shortened life span of adults after metamorphosis for both males and females, whereas control and numerical density reduction “removals” treatments lengthened life span. Our results show strong lethal and nonlethal outcomes of the combination on Ae. aegypti. These findings suggest an additional benefit, decreases adult life span, of the use of an IGR when combined with a natural predator of mosquitoes that may be exploited to improve mosquito control strategies to reduce the risk of disease transmission.

Aedes mosquito responses to control interventions against the Chikungunya outbreak of Dire Dawa, Eastern Ethiopia

Aedes mosquito responses to control interventions against the Chikungunya outbreak of Dire Dawa, Eastern Ethiopia

Acoustotactic response of mosquitoes in untethered flight to incidental sound

Mosquitoes are vectors for some of the most devastating diseases on the planet. Given the centrality of acoustic sensing in the precopulatory behavior of these vectors, the use of an exogenous acoustic stimulus offers the potential of interfering with the courtship behavior of these insects. Previous research on the acoustotactic response of mosquitoes has been conducted on tethered preparations using low-intensity sound stimuli. To quantify differences in acoustotactic responses between mosquitos of distinct sex and species, we examined the effects of incidental sound stimuli on the flight behavior of free-flying male vs. female Aedes aegypti and Anopheles gambiae mosquitoes. The key variables were sound frequency (100–1000 Hz) and intensity (67–103 dB, measured at 12.5 cm from the source), and the acoustotactic response was measured in terms of the relative increase in flight speed in response to the stimulus. The data show, for the first time, significant sex- and species-specific differences in acoustotactic responses. A. aegypti exhibited a greater response to sound stimulus compared to An. gambiae, and the response also extended over a larger range of frequencies. Furthermore, the males of both species displayed a greater acoustotactic response than females, with An. gambiae females exhibiting minimal response to sound.

Genetic changes of Plasmodium vivax tempers host tissue-specific responses in Anopheles stephensi

Recently, we showed how an early restriction of gut flora proliferation by Plasmodium vivax favors immune-suppression and Plasmodium survival in the gut lumen (Sharma et al., 2020). Here, we asked post gut invasion how P. vivax interacts with individual tissues such as the midgut, hemocyte, and salivary glands, and manages its survival in the mosquito host. Our data from tissue-specific comparative RNA-Seq analysis and extensive temporal/spatial expression profiling of selected mosquito transcripts in the uninfected and P. vivax infected mosquito’s tissues indicated that (i) a transient suppression of gut metabolic machinery by early oocysts; (ii) enriched expression of nutritional responsive proteins and immune proteins against late oocysts, together may ensure optimal parasite development and gut homeostasis restoration; (iii) pre-immune activation of hemocyte by early gut-oocysts infection via REL induction (p ​< ​0.003); and altered expression of hemocyte-encoded immune proteins may cause rapid removal of free circulating sporozoites from hemolymph; (iv) while a strong suppression of salivary metabolic activities, and elevated expression of salivary specific secretory, as well as immune proteins together, may favor the long-term storage and survival of invaded sporozoites. Finally, our RNA-Seq-based discovery of 4449 transcripts of Plasmodium vivax origin, and their developmental stage-specific expression modulation in the corresponding infected mosquito tissues, predicts a possible mechanism of mosquito responses evasion by P. vivax. Conclusively, our system-wide RNA-Seq analysis provides the first genetic evidence of direct mosquito-Plasmodium interaction and establishes a functional correlation.

Prediction of neuropeptide precursors and differential expression of adipokinetic hormone/corazonin-related peptide, hugin and corazonin in the brain of malaria vector Nyssorhynchus albimanus during a Plasmodium berghei infection.

Insect neuropeptides, play a central role in the control of many physiological processes. Based on an analysis of Nyssorhynchus albimanus brain transcriptome a neuropeptide precursor database of the mosquito was described. Also, we observed that adipokinetic hormone/corazonin-related peptide (ACP), hugin and corazonin encoding genes were differentially expressed during Plasmodium infection. Transcriptomic data from Ny. albimanus brain identified 29 pre-propeptides deduced from the sequences that allowed the prediction of at least 60 neuropeptides. The predicted peptides include isoforms of allatostatin C, orcokinin, corazonin, adipokinetic hormone (AKH), SIFamide, capa, hugin, pigment-dispersing factor, adipokinetic hormone/corazonin-related peptide (ACP), tachykinin-related peptide, trissin, neuropeptide F, diuretic hormone 31, bursicon, crustacean cardioactive peptide (CCAP), allatotropin, allatostatin A, ecdysis triggering hormone (ETH), diuretic hormone 44 (Dh44), insulin-like peptides (ILPs) and eclosion hormone (EH). The analysis of the genome of An. albimanus and the generated transcriptome, provided evidence for the identification of myosuppressin neuropeptide precursor. A quantitative analysis documented increased expression of precursors encoding ACP peptide, hugin and corazonin in the mosquito brain after Plasmodium berghei infection. This work represents an initial effort to characterize the neuropeptide precursors repertoire of Ny. albimanus and provides information for understanding neuroregulation of the mosquito response during Plasmodium infection.

Behavioral response of insecticide-resistant mosquitoes against spatial repellent: A modified self-propelled particle model simulation.

Rapidly increasing pyrethroid insecticide resistance and changes in vector biting and resting behavior pose serious challenges in malaria control. Mosquito repellents, especially spatial repellents, have received much attention from industry. We attempted to simulate interactions between mosquitoes and repellents using a machine learning method, the Self-Propelled Particle (SPP) model, which we modified to include attractiveness/repellency effects. We simulated a random walk scenario and scenarios with insecticide susceptible/resistant mosquitoes against repellent alone and against repellent plus attractant (to mimic a human host). Simulation results indicated that without attractant/repellent, mosquitoes would fly anywhere in the cage at random. With attractant, all mosquitoes were attracted to the source of the odor by the end. With repellent, all insecticide-susceptible mosquitoes eventually moved to the corner of the cage farthest from the repellent release point, whereas, a high proportion of highly resistant mosquitoes might reach the attractant release point (the human) earlier in the simulation. At fixed concentration, a high proportion of mosquitoes could be able to reach the host when the relative repellency efficacy (compare to attractant efficacy) was <1, whereas, no mosquitoes reached the host when the relative repellency efficacy was > 1. This result implies that repellent may not be sufficient against highly physiologically insecticide resistant mosquitoes, since very high concentrations of repellent are neither practically feasible nor cost-effective.

Mosquito defensin facilitates Japanese encephalitis virus infection by downregulating the C6/36 cell-surface antiviral protein HSC70B

Japanese encephalitis virus (JEV) is a viral zoonosis that can cause viral encephalitis, death and disability whose primary vector is the Culex mosquito. Viral infection induces a series of antimicrobial peptide responses in mosquitoes, and the effector defensin enhances JEV replication in mosquitoes. However, the underlying mechanisms by which defensin enhances JEV are not fully understood. Here, we found that mosquito defensin could downregulate the antiviral protein HSC70B and enhance virus infection in mosquitoes. The cell-surface protein HSC70B was significantly downregulated by JEV infection and defensin treatment. Low levels of HSC70B were beneficial to JEV infection in mosquitoes. Taken together, these findings show that defensin and HSC70B axis facilitates JEV infection in the mosquito.

The Differential Metabolic Profiles Between Deltamethrin-Resistant and -Susceptible Strains of Aedes albopictus (Diptera: Culicidae) by 1H-NMR.

Metabolomics can indicate the physiological and biochemical responses of mosquitoes to different stimulants, including insecticides, which allow them to adapt to different inhospitable environments. Though metabolic differences between insecticide-resistant and -susceptible strains have been established for other mosquito species, such as Anopheles and Culex, it is yet to be done for Aedes albopictus (Skuse). In this study, nuclear magnetic resonance (NMR) spectroscopy-based metabolomic analysis performed on Ae. albopictus deltamethrin-resistant and -susceptible strains showed significant differences in amino acid, organic acid, and sugar metabolism. Concentrations of neutral amino acids and sugars tended to be lower in the deltamethrin-resistant strain than in the deltamethrin-suceptible strain, but the concentration of basic and acidic amino acids and organic acids increased. All these changes might accommodate biochemical and physiological needs in deltamethrin-resistant mosquitoes, such as enzyme synthesis and detoxification. This was further confirmed by the predictable draft metabolic map. This is the first report using NMR spectroscopy to investigate the metabolic differences between deltamethrin-resistant and -susceptible strains of Ae. albopictus. To a certain degree, this demonstrates how Ae. albopictus develop insecticide resistance by metabolic reprograming to survive under the insecticide pressure.

Antifungal immune responses in mosquitoes (Diptera: Culicidae): A review

Mosquitoes transmit many parasites and pathogens to humans that cause significant morbidity and mortality. As such, we are constantly looking for new methods to reduce mosquito populations, including the use of effective biological controls. Entomopathogenic fungi are excellent candidate biocontrol agents to control mosquitoes. Understanding the complex ecological, environmental, and molecular interactions between hosts and pathogens are essential to create novel, effective and safe biocontrol agents. Understanding how mosquitoes recognize and eliminate pathogens such as entomopathogenic fungi may allow us to create insect-order specific biocontrol agents to reduce pest populations. Here we summarize the current knowledge of fungal infection, colonization, development, and replication within mosquitoes and the innate immune responses of the mosquitoes towards the fungal pathogens, emphasizing those features required for an effective mosquito biocontrol agent.

Zika Virus Infection Results in Biochemical Changes Associated With RNA Editing, Inflammatory and Antiviral Responses in Aedes albopictus.

Rapid and significant range expansion of both the Zika virus (ZIKV) and its Aedes vector species has resulted in the declaration of ZIKV as a global health threat. Successful transmission of ZIKV by its vector requires a complex series of interactions between these entities including the establishment, replication and dissemination of the virus within the mosquito. The metabolic conditions within the mosquito tissues play a critical role in mediating the crucial processes of viral infection and replication and represent targets for prevention of virus transmission. In this study, we carried out a comprehensive metabolomic phenotyping of ZIKV infected and uninfected Ae. albopictus by untargeted analysis of primary metabolites, lipids and biogenic amines. We performed a comparative metabolomic study of infection state with the aim of understanding the biochemical changes resulting from the interaction between the ZIKV and its vector. We have demonstrated that ZIKV infection results in changes to the cellular metabolic environment including a significant enrichment of inosine and pseudo-uridine (Ψ) levels which may be associated with RNA editing activity. In addition, infected mosquitoes demonstrate a hypoglycemic phenotype and show significant increases in the abundance of metabolites such as prostaglandin H2, leukotriene D4 and protoporphyrinogen IX which are associated with antiviral activity. These provide a basis for understanding the biochemical response to ZIKV infection and pathology in the vector. Future mechanistic studies targeting these ZIKV infection responsive metabolites and their associated biosynthetic pathways can provide inroads to identification of mosquito antiviral responses with infection blocking potential.

Landing mosquitoes bounce when engaging a substrate

In this experimental study we film the landings of Aedes aegypti mosquitoes to characterize landing behaviors and kinetics, limitations, and the passive physiological mechanics they employ to land on a vertical surface. A typical landing involves 1–2 bounces, reducing inbound momentum by more than half before the mosquito firmly attaches to a surface. Mosquitoes initially approach landing surfaces at 0.1–0.6 m/s, decelerating to zero velocity in approximately 5 ms at accelerations as high as 5.5 gravities. Unlike Dipteran relatives, mosquitoes do not visibly prepare for landing with leg adjustments or body pitching. Instead mosquitoes rely on damping by deforming two forelimbs and buckling of the proboscis, which also serves to distribute the impact force, lessening the potential of detection by a mammalian host. The rebound response of a landing mosquito is well-characterized by a passive mass-spring-damper model which permits the calculation of force across impact velocity. The landing force of the average mosquito in our study is approximately 40 upmuN corresponding to an impact velocity of 0.24 m/s. The substrate contact velocity which produces a force perceptible to humans, 0.42 m/s, is above 85% of experimentally observed landing speeds.