Host Seeking Research Articles

Genome report: First whole genome sequence of Triatoma sanguisuga (Le Conte, 1855), vector of Chagas disease.

Triatoma sanguisuga is the most widespread triatomine bug species in the United States (US). The species vectors the human parasite Trypanosoma cruzi , which causes Chagas disease. Vector-borne Chagas disease is rarely diagnosed in the US, but T. sanguisuga has been implicated in a handful of cases. Despite its public health importance, little is known about the genomics or population genetics of T. sanguisuga . Here, we used long-read sequencing to assemble the first whole genome sequence for T. sanguisuga using DNA extracted from one adult specimen from Delaware. The final size of the genome was 1.162 Gbp with 77.7x coverage. The assembly consisted of 183 contigs with an N50 size of 94.97 Kb. The Benchmarking Universal Single-Copy Ortholog (BUSCO) complete score was 99.1%, suggesting a very complete assembly. Genome-wide GC level was 33.56%, and DNA methylation was 18.84%. The genome consists of 61.4% repetitive DNA and 17,799 predicted coding genes. The assembled T. sanguisuga genome was slightly larger than that of Triatominae species Triatoma infestans and Rhodnius prolixus (949 Mbp with 90.4% BUSCO score and 706 Mbp with 96.5% BUSCO score, respectively). The T. sanguisuga genome is the first North American triatomine species genome to be sequenced, and it is the most complete genome yet for any Triatominae species. The T. sangsuiga genome will allow for deeper investigations into epidemiologically relevant aspects of this important vector species, including blood feeding, host seeking, and parasite competence, thus providing potential vector-borne disease management targets and strengthening public health preparedness.

Invasive brown algae (Sargassum spp.) as a potential source of biocontrol against Aedes aegypti

Influxes of sargassos are responsible for economic and environmental disasters in areas where they bloom, especially in regions whose main income relies on tourism and with limited capacity for sanitation and public health response. A promising way of valorization would be to convert this incredible biomass into tools to fight the deadly vector mosquito Aedes aegypti. In the present study, we generated hydrolates and aqueous extracts from three main Sargassum morphotypes identified in Guadeloupe (French West Indies): Sargassum natans VIII, Sargassum natans I and Sargassum fluitans. We conducted a chemical characterization and a holistic evaluation of their potential to induce toxic and behavioral effects in Ae. aegypti. Despite the low insecticidal potential observed for all the extracts, we found that S. natans VIII and S. fluitans hydrolates deterred oviposition, induced contact irritancy and stimulated blood feeding behavior in host seeking Ae. aegypti females, while aqueous extracts from S. natans I and S. fluitans deterred both blood feeding behavior and oviposition. Chemical characterization evidenced the presence of phenylpropanoid, polyphenols, amino acids and esters. Thus, Sargassum spp. aqueous extracts and hydrolates could be used to manipulate Ae. aegypti behavior and be valorized as control tools against this mosquito.

The potential role of the Asian bush mosquito Aedes japonicus as spillover vector for West Nile virus in the Netherlands

BackgroundIn recent years the Asian bush mosquito Aedes japonicus has invaded Europe, including the Netherlands. This species is a known vector for a range of arboviruses, possibly including West Nile virus (WNV). As WNV emerged in the Netherlands in 2020, it is important to investigate the vectorial capacity of mosquito species present in the Netherlands to estimate the risk of future outbreaks and further spread of the virus. Therefore, this study evaluates the potential role of Ae. japonicus in WNV transmission and spillover from birds to dead-end hosts in the Netherlands.MethodsWe conducted human landing collections in allotment gardens (Lelystad, the Netherlands) in June, August and September 2021 to study the diurnal and seasonal host-seeking behaviour of Ae. japonicus. Furthermore, their host preference in relation to birds using live chicken-baited traps was investigated. Vector competence of field-collected Ae. japonicus mosquitoes for two isolates of WNV at two different temperatures was determined. Based on the data generated from these studies, we developed a Susceptible-Exposed-Infectious-Recovered (SEIR) model to calculate the risk of WNV spillover from birds to humans via Ae. japonicus, under the condition that the virus is introduced and circulates in an enzootic cycle in a given area.ResultsOur results show that Ae. japonicus mosquitoes are actively host seeking throughout the day, with peaks in activity in the morning and evening. Their abundance in August was higher than in June and September. For the host-preference experiment, we documented a small number of mosquitoes feeding on birds: only six blood-fed females were caught over 4 full days of sampling. Finally, our vector competence experiments with Ae. japonicus compared to its natural vector Culex pipiens showed a higher infection and transmission rate when infected with a local, Dutch, WNV isolate compared to a Greek isolate of the virus. Interestingly, we also found a small number of infected Cx. pipiens males with virus-positive leg and saliva samples.ConclusionsCombining the field and laboratory derived data, our model predicts that Ae. japonicus could act as a spillover vector for WNV and could be responsible for a high initial invasion risk of WNV when present in large numbers.Graphical

Decoding the genome of bloodsucking midge Forcipomyia taiwana (diptera: Ceratopogonidae): Insights into odorant receptor expansion

Biting midges, notably those within the Ceratopogonidae family, have long been recognized for their epidemiological significance, both as nuisances and vectors for disease transmission in vertebrates. Despite their impact, genomic insights into these insects, particularly beyond the Culicoides genus, remain limited. In this study, we assembled the Forcipomyia taiwana (Shiraki) genome, comprising 113 scaffolds covering 130.4 Mbps—with the longest scaffold reaching 7.6 Mbps and an N50 value of 2.6 Mbps—marking a pivotal advancement in understanding the genetic architecture of ceratopogonid biting midges. Phylogenomic analyses reveal a shared ancestry between F. taiwana and Culicoides sonorensis Wirth & Jones, dating back approximately 124 million years, and highlight a dynamic history of gene family expansions and contractions within the Ceratopogonidae family. Notably, a substantial expansion of the odorant receptor (OR) gene family was observed, which is crucial for the chemosensory capabilities that govern biting midges' interactions with their environment, including host seeking and oviposition behaviors. The distribution of OR genes across the F. taiwana genome displays notable clusters on scaffolds, indicating localized tandem gene duplication events. Additionally, several collinear regions were identified, hinting at segmental duplications, inversions, and translocations, contributing to the olfactory system's evolutionary complexity. Among the 156 ORs identified in F. taiwana, 134 are biting midge-specific ORs, distributed across three distinct clades, each exhibiting unique motif features that distinguish them from the others. Through weighted gene co-expression network analysis, we correlated distinct gene modules with sex and reproductive status, laying the groundwork for future investigations into the interplay between gene expression and adaptive behaviors in F. taiwana. In conclusion, our study not only highlights the unique olfactory repertoire of ceratopogonid biting midges but also sets the stage for future studies into the genetic underpinnings of their unique biological traits and ecological strategies.

Optimizing malaria vector control in the Greater Mekong Subregion: a systematic review and mathematical modelling study to identify desirable intervention characteristics

BackgroundIn the Greater Mekong Subregion (GMS), new vector-control tools are needed to target mosquitoes that bite outside during the daytime and night-time to advance malaria elimination.MethodsWe conducted systematic literature searches to generate a bionomic dataset of the main malaria vectors in the GMS, including human blood index (HBI), parity proportion, sac proportion (proportion with uncontracted ovary sacs, indicating the amount of time until they returned to host seeking after oviposition) and the resting period duration. We then performed global sensitivity analyses to assess the influence of bionomics and intervention characteristics on vectorial capacity.ResultsOur review showed that Anopheles minimus, An. sinensis, An. maculatus and An. sundaicus display opportunistic blood-feeding behaviour, while An. dirus is more anthropophilic. Multivariate regression analysis indicated that environmental, climatic and sampling factors influence the proportion of parous mosquitoes, and resting duration varies seasonally. Sensitivity analysis highlighted HBI and parity proportion as the most influential bionomic parameters, followed by resting duration. Killing before feeding is always a desirable characteristic across all settings in the GMS. Disarming is also a desirable characteristic in settings with a low HBI. Repelling is only an effective strategy in settings with a low HBI and low parity proportion. Killing after feeding is only a desirable characteristic if the HBI and parity proportions in the setting are high.ConclusionsAlthough in general adopting tools that kill before feeding would have the largest community-level effect on reducing outdoor transmission, other modes of action can be effective. Current tools in development which target outdoor biting mosquitoes should be implemented in different settings dependent on their characteristics.Graphical

Behavioral responses of Ixodiphagus hookeri (Hymenoptera; Encyrtidae) to Rhipicephalus sanguineus nymphs (Ixodida: Ixodidae) and dog hair volatiles

Abstract The wasp Ixodiphagus hookeri (Howard; Hymenoptera: Encyrtidae) parasitizes the larvae and nymphs of several tick species worldwide. In this study, we investigated whether I. hookeri females are attracted to Rhipicephalus sanguineus Latreille (Ixodida: Ixodidae) nymphs, hair samples from non-infested and infested Schnauzer dog volatiles, and a synthetic blend of compounds isolated from dog hair in a Y-tube glass olfactometer. We also studied the acceptance of fed and unfed R. sanguineus nymphs by female wasps. Our results showed that I. hookeri females were not attracted to R. sanguineus nymphs. In contrast, they were attracted to dog hair samples and extracts from non-infested and R. sanguineus-infested Schnauzer dog hair. Wasps were more attracted to hair samples from R. sanguineus-infested dogs than to those from non-infested dogs. In addition, I. hookeri females were attracted to a synthetic blend composed of hexanal, heptanal, and isovaleric acid, compounds previously isolated from dog hair. In contrast, I. hookeri females were not attracted to binary blends or individual compounds. Female wasps showed a higher acceptance of live-fed and live-unfed R. sanguineus nymphs than of dead fed nymphs washed with dichloromethane. Our results suggests that the host seeking behavior of I. hookeri females is attributable to a blend of volatiles from infested and non-infested schnauzer dogs, used to attack R. sanguineus nymphs.

An update and review of arthropod vector sensory systems: Potential targets for behavioural manipulation by parasites and other disease agents.

For over a century, vector ecology has been a mainstay of vector-borne disease control. Much of this research has focused on the sensory ecology of blood-feeding arthropods (black flies, mosquitoes, ticks, etc.) with terrestrial vertebrate hosts. Of particular interest are the cues and sensory systems that drive host seeking and host feeding behaviours as they are critical for a vector to locate and feed from a host. An important yet overlooked component of arthropod vector ecology are the phenotypic changes observed in infected vectors that increase disease transmission. While our fundamental understanding of sensory mechanisms in disease vectors has drastically increased due to recent advances in genome engineering, for example, the advent of CRISPR-Cas9, and high-throughput "big data" approaches (genomics, proteomics, transcriptomics, etc.), we still do not know if and how parasites manipulate vector behaviour. Here, we review the latest research on arthropod vector sensory systems and propose key mechanisms that disease agents may alter to increase transmission.

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.

AsOBP1 is required for host seeking in the malaria vector mosquito, Anopheles sinensis

AsOBP1 is required for host seeking in the malaria vector mosquito, Anopheles sinensis

Pyrethroid-treated bed nets impair blood feeding performance in insecticide resistant mosquitoes

The blood feeding performance of female mosquitoes directly impacts their ability to transmit malaria. Yet their host seeking and blood feeding behaviours in the presence of insecticide-treated nets (ITNs) are still poorly understood. This work explores how both insecticide resistant and susceptible Anopheles gambiae s.l. mosquitoes interact with pyrethroid nets (PermaNet 2.0 or Olyset net) or an untreated net (UTN) while attempting to blood feed on a human arm. Regardless of mosquito resistance status, the ITNs did not efficiently prevent host searching but reduced blood feeding success by 34.1 (29.31–38.95) %. The Permanet and Olyset net reduced to 227.5 (208.19–246.77) sec and 235.9 (214.03–257.74) sec the average blood feeding duration from 369.9 (342.78–397.04) sec with the UTN. The ingested blood volume was on average 22% lower for all mosquitoes exposed to insecticide. When feeding through ITNs, the blood volume flow rate of the susceptible strain increased by 35%, but no significant difference was found in the resistant strain. Thus, whilst the presence of the insecticide in ITNs reduced mosquito blood feeding success and blood volume, the mosquito’s ability to respond by accelerating her rate of blood ingestion may further reduce the impact of ITNs on resistant mosquitoes.

Effect of Temperature on Mosquito Olfaction.

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

Horse fly and deer fly (Diptera: Tabanidae) species and abundance following logging in northwestern Ontario

Horse flies and deer flies are important components of habitat for many species, but how logging may alter the communities of biting flies in the boreal forest is largely unexplored. Such knowledge is important to conservation, because biting flies can shape the behaviour and sometimes even the demography of vertebrates in the boreal forest. We compared the abundance and distribution of Tabanidae species in managed boreal forest stands in northwestern Ontario with three harvest histories. Using sweep nets during 2 summers, we collected specimens from forest stands that were young (harvested 20–35 years ago), intermediate (harvested 36–69 years ago), and old (unharvested or harvested ≥ 70 years ago). We tested the hypothesis that greater harassment potential in young stands was due not only to more abundant tabanids, but more abundant obligate blood-feeding (anautogenous) tabanids, which are more persistent in host seeking. We caught 8,895 specimens from 44 species. The oldest stands had the highest species richness, but the lowest abundance of individuals, whereas young stands had the highest relative abundance and the most anautogenous specimens, potentially causing more stress to hosts in young stands. These results demonstrate that forest harvesting may reduce habitat suitability for many large mammals due to an altered tabanid community.

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.

LABORATORY EVALUATION OF BIGSHOT MAXIM, REPELCARE, AND CLOVE ESSENTIAL OIL, SYZYGIUM AROMATICUM AGAINST THE LONE STAR TICK, AMBLYOMMA AMERICANUM

Synthetic acaricides have been the most used method for controlling tick populations. However, their frequent application has had negative impacts on treated animals and the environment and led to the development of resistance in tick populations. These factors generated interest and the need to find more environmentally friendly alternatives. In this study, the repellency and acaricidal effects of BigShot Maxim (AIs: cedarwood oil, cinnamon oil, thyme oil), RepelCare (AIs: turmeric oil and eucalyptus oil), and clove oil, Syzygium aromaticum L., were tested against the lone star tick, Amblyomma Americanum (Linnaeus). Despite no evidence of repellency for BigShot Maxim, RepelCare and clove oil, its disruption of the host seeking behavior after contact to the products was observed and further investigated. BigShot Maxim and clove oil were selected for mortality testing and resulted in complete mortality of male, female ticks, and nymphs with some application rates. The results from this study provide a better understanding of repellency and acaricidal effects of botanical products against the lone star tick that can be used to further improve the development of green chemistry. However, further studies are warranted before these botanical products can be implemented as effective alternatives to chemical acaricides to use in integrated vector control.

Thermal preference of adult mosquitoes (Culicidae) and biting midges (Ceratopogonidae) at different altitudes in Switzerland.

Mosquitoes (Diptera: Culicidae) and biting midges (Diptera: Ceratopogonidae) are among the most important vectors of human and veterinary pathogens. For modelling the distribution of these pathogens, entomological aspects are essential, which in turn are highly dependent on environmental factors, such as temperature. In this study, mosquitoes and biting midges were sampled in multiple microclimates at two low (360, 480 meters above sea level, m.a.s.l.) and two high (1250, 1530 m.a.s.l.) altitude locations in Switzerland. Sets of various traps (CO2 -baited CDC, LED-UV, resting boxes, oviposition cups) equipped with dataloggers were placed in transects at five sites with similar vegetation at each location. Only the CDC and the LED-UV traps collected enough insects for analyses. Taxonomic diversity was greater for mosquitoes but lower for biting midges at lower altitudes. Both mosquitoes and biting midges had a thermal preference. Culicoides preferred the traps with warmer microclimate, especially at lower altitudes, whereas mosquito preferences depended on the species, but not on altitude. Relative humidity had a significant positive impact on catches of biting midges but not mosquitoes. To obtain better data on thermal preferences of resting and ovipositing vectors in addition to host seeking individuals, new and improved collecting methods are needed.

Entomological Surveillance Planning Tool (ESPT)-generated actionable evidence on human and vector behaviours optimizes present interventions and reduces exposure to Anopheles vectors in two communities of Guna Yala, Panamá

BackgroundAlthough most of Panamá is free from malaria, localized foci of transmission persist, including in the Guna Yala region. Government-led entomological surveillance using an Entomological Surveillance Planning Tool (ESPT) sought to answer programmatically relevant questions on local entomological drivers of transmission and gaps in protection to guide local vector control decision-making.MethodsThe ESPT was used to design a sampling plan to answer priority programmatic questions about the appropriateness of Long Lasting Insecticidal Nets (LLINs) and spaces and times where humans remain exposed to Anopheles bites (gaps in protection) in the communities of Permé and Puerto Obaldía, Guna Yala. Adult Anopheles were sampled at three time points via human landing catches (HLCs) during the rainy and dry seasons (2018/2019). Human behaviour observations (HBOs) were conducted alongside HLCs to examine intervention use, indoor versus outdoor activity, and sleeping patterns. HLC and HBO data were integrated to evaluate HBO-adjusted human biting rate (HBR).ResultsA total of 7,431 adult Anopheles were collected across both sites. Of the 450 specimens molecularly confirmed to species-level, 75.5% (n = 340) were confirmed as Anopheles Nyssorhynchus albimanus, followed by Anopheles (Ny.) aquasalis. Anopheles host seeking activity was demonstrated to be primarily exophagic throughout all sampling periods and in both communities. When adjusted with HBOs, exposure to mosquito bites was predominantly indoors and overnight in Permé (Nov, Mar), compared to predominantly outdoors in Puerto Obaldía (Nov, Mar, Jul). Differences in site-specific human-vector exposure profiles were due to contrasting cultural and lifestyle practices between Permé and Puerto Obaldía (possibly partly influenced by the absence of electricity in Permé), and lower LLIN use in Permé. This evidence supported a previously planned LLIN campaign alongside a social behaviour change communication (SBCC) strategy in the Guna Yala Comarca (Jul 2019), which increased LLIN use. In turn, this led to a reduction of indoor exposure to mosquito bites, and a shift to predominant outdoor exposure to mosquito bites.ConclusionESPT-based question-driven planning and the integration of HBOs, intervention, and HLC data generated evidence towards answering the programmatic questions. This evidence enabled the characterization of site-specific human-vector exposure profiles, and the quantification of remaining gaps in protection. These data also provide important insights into remaining gaps in protection that must be addressed to further reduce human exposure to mosquito bites at these sites.

Antennal and maxillary palp morphology, and sensillar equipment, of the spruce bark beetle predators, Medetera signaticornis and Medetera infumata (Diptera: Dolichopodidae)

Many long-legged Medetera flies are natural enemies of bark beetle pests, which they detect using olfactory cues, likely through olfactory sensilla on the antennae and maxillary palps. Morphological characterisation of olfactory sensilla among insects can provide a basis for future taxonomic, phylogenetic or electrophysiological studies. Scanning electron microscopy was used to describe the morphology of olfactory organs and sensillar equipment of Medetera signaticornis and M. infumata. Three different olfactory sensillum types were found in both fly species, sensilla trichodea, s. basiconica and grooved pegs. Based on size and wall structure, s. trichodea and s. basiconica were categorised into different subtypes. Sharp-tipped curved s. trichodea, and small, large and thin s. basiconica were found on the antennal postpedicel of M. signaticornis adults, while grooved s. basiconica were found in M. infumata. The density of sharp-tipped long s. trichodea was significantly higher in males compared to females, and in M. signaticornis compared to M. infumata. Long-grooved s. basiconica were found grouped in a small pit on the maxillary palps of both species. Comparison of our results with the limited available ecological data suggests that differences in numbers of specific sensillum types may reflect adaptations related to olfactory-driven behaviours such as host seeking.

Human and vector behaviors determine exposure to Anopheles in Namibia

BackgroundAlthough the Republic of Namibia has significantly reduced malaria transmission, regular outbreaks and persistent transmission impede progress towards elimination. Towards an understanding of the protective efficacy, as well as gaps in protection, associated with long-lasting insecticidal nets (LLINs), human and Anopheles behaviors were evaluated in parallel in three malaria endemic regions, Kavango East, Ohangwena and Zambezi, using the Entomological Surveillance Planning Tool to answer the question: where and when are humans being exposed to bites of Anopheles mosquitoes?MethodsSurveillance activities were conducted during the malaria transmission season in March 2018 for eight consecutive nights. Four sentinel structures per site were selected, and human landing catches and human behavior observations were consented to for a total of 32 collection nights per site. The selected structures were representative of local constructions (with respect to building materials and size) and were at least 100 m from each other. For each house where human landing catches were undertaken, a two-person team collected mosquitoes from 1800 to 0600 hours.ResultsSurveillance revealed the presence of the primary vectors Anopheles arabiensis, Anopheles gambiae sensu stricto (s.s.) and Anopheles funestus s.s., along with secondary vectors (Anopheles coustani sensu lato and Anopheles squamosus), with both indoor and outdoor biting behaviors based on the site. Site-specific human behaviors considerably increased human exposure to vector biting. The interaction between local human behaviors (spatial and temporal presence alongside LLIN use) and vector behaviors (spatial and temporal host seeking), and also species composition, dictated where and when exposure to infectious bites occurred, and showed that exposure was primarily indoors in Kavango East (78.6%) and outdoors in Ohangwena (66.7%) and Zambezi (81.4%). Human behavior-adjusted exposure was significantly different from raw vector biting rate.ConclusionsIncreased LLIN use may significantly increase protection and reduce exposure to malaria, but may not be enough to eliminate the disease, as gaps in protection will remain both indoors (when people are awake and not using LLINs) and outdoors. Alternative interventions are required to address these exposure gaps. Focused and question-based operational entomological surveillance together with human behavioral observations may considerably improve our understanding of transmission dynamics as well as intervention efficacy and gaps in protection.Graphical

Piercing and sucking mouth parts sensilla of irradiated mosquito, Culex pipiens (Diptera: Culicidae) with gamma radiation

Morphology, distribution and function of the sensilla associated with mouthparts of female mosquito, Culex pipiens (Diptera: Culicidae) were studied by using the scanning electron microscope. The female mosquito mouthpart is a piercing and sucking type and carrying many sensory organs which plays major role in host seeking, and oviposition behaviour of mosquitoes. Six types of mouthpart sensilla have been identified and described, Sensilla trichoidea, chaetica, basiconica, Capitate peg, campaniformia and squamiformia. Females mosquito were irradiated as pupae with 20, 40 and 80 Gy of gamma radiation to investigate their effect on mouthparts sensilla. Dose of 20 Gy had slight effect on the different types of the sensilla as compared with the control. Features of malformation observed in the maxillary palp segments at this radiation dose include falling of some scales (sensilla squamiformia) at certain parts of the 1st segment leaving vacuoles. At 40 Gy, the maxillary palp segments were liquefied in some areas leaving undistinguished shape. At the highest dose 80 Gy, the tip of labial palps had many tears and showed were most affected, especially, at the trichoid, basiconic and chateica sensilla. All these malformations may lead the mosquitoes to not reaching the host, impeding their ability to transmit diseases or killing the mosquitoes, and this is the main objective of this study.

Using next generation sequencing for molecular detection and differentiation of Anaplasma phagocytophilum variants from host seeking Ixodes scapularis ticks in the United States

Anaplasmosis is increasingly common in the United States, with cases being reported over an expanding geographic area. To monitor for changes in risk of human infection, the U.S. Centers for Disease Control and Prevention monitors the distribution and abundance of host-seeking vector ticks (Ixodes scapularis and Ixodes pacificus) and their infection with Anaplasma phagocytophilum. While several variants of A. phagocytophilum circulate in I. scapularis, only the human-active variant (Ap-ha) appears to be pathogenic in humans. Failure to differentiate between human and non-human variants may artificially inflate estimates of the risk of human infection. Efforts to differentiate the Ap-ha variant from the deer variant (Ap-V1) in ticks typically rely on traditional PCR assays coupled with sequencing of PCR products. However, laboratories are increasingly turning to Next Generation Sequencing (NGS) to increase testing efficiency, retain high sensitivity, and increase specificity compared with traditional PCR assays. We describe a new NGS assay with novel targets that accurately segregate the Ap-ha variant from other non-human variants and further identify unique clades within the human and non-human variants. Recognizing that not all investigators have access to NGS technology, we also developed a PCR assay based on one of the novel targets so that variants can be visualized using agarose gel electrophoresis without the need for subsequent sequencing. Such an assay may be used to improve estimates of human risk of developing anaplasmosis in North America.