Blacklegged Tick Ixodes Scapularis Research Articles

Effects of residential acaricide treatments on patterns of pathogen coinfection in blacklegged ticks.

Medically important ixodid ticks often carry multiple pathogens, with individual ticks frequently coinfected and capable of transmitting multiple infections to hosts, including humans. Acquisition of multiple zoonotic pathogens by immature blacklegged ticks (Ixodes scapularis) is facilitated when they feed on small mammals, which are the most competent reservoir hosts for Anaplasma phagocytophilum (which causes anaplasmosis in humans), Babesia microti (babesiosis) and Borrelia burgdorferi (Lyme disease). Here, we used data from a large-scale, long-term experiment to ask whether patterns of single and multiple infections in questing nymphal I. scapularis ticks from residential neighbourhoods differed from those predicted by independent assortment of pathogens, and whether patterns of coinfection were affected by residential application of commercial acaricidal products. Quantitative polymerase chain reaction was used for pathogen detection in multiplex reactions. In control neighbourhoods and those treated with a fungus-based biopesticide deployed against host-seeking ticks (Met52), ticks having only single infections of either B. microti or B. burgdorferi were significantly less common than expected, whereas coinfections with these 2 pathogens were significantly more common. However, use of tick control system bait boxes, which kill ticks attempting to feed on small mammals, eliminated the bias towards coinfection. Although aimed at reducing the abundance of host-seeking ticks, control methods directed at ticks attached to small mammals may influence human exposure to coinfected ticks and the probability of exposure to multiple tick-borne infections.

Partial trailside Japanese barberry (Ranunculales: Berberidaceae) removal did not reduce the abundance of questing blacklegged ticks (Acari: Ixodidae).

In a nature reserve in southern Maine, we removed invasive Japanese barberry (Berberis thunbergii de Candolle) along sections of forested recreational trails that ran through dense barberry infestations. Barberry thickets provide questing substrate and a protective microclimate for blacklegged ticks (Ixodes scapularis Say), and trail users could brush up against encroaching barberry and acquire ticks. Trailside barberry removal will reduce or eliminate encroaching tick questing substrate and could reduce trailside questing tick abundance by creating a microclimate more hostile to ticks. The same-day cut-and-spray treatment comprised mechanical cutting of barberry clumps (individual plants with numerous ramets) followed immediately by targeted herbicide application to the resulting root crowns. The treatment created trail shoulders to a lateral width of 1-2 m on both sides of 100-m trail sections, with initial treatment in the fall of 2013 and one retreatment in the summer of 2014. Our aim was to remove 90% of barberry clumps to achieve a 50% or better reduction in questing tick abundance on trail shoulders. However, by the fall of 2015, there were only 41% fewer barberry clumps on treated vs. untreated trail sections and there was no reduction in either adults or nymphs. We concluded that our barberry treatment protocol was not sufficiently aggressive since the resulting ecotone habitat on trail shoulders proved suitable for questing I. scapularis. In principle, cutting back barberry along trails should reduce trail user contact with questing deer ticks, but we were unable to demonstrate a reduction in trailside tick abundance.

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.

Genetic manipulation of an Ixodes scapularis cell line.

Although genetic manipulation is one of the hallmarks of model organisms, its applicability to non-model species has remained difficult due to our limited understanding of their fundamental biology. For instance, manipulation of a cell line originated from the black-legged tick Ixodes scapularis, an arthropod that serves as a vector for several human pathogens, has yet to be established. Here, we demonstrate the successful genetic modification of the commonly used tick ISE6 line through ectopic expression and clustered regularly interspaced palindromic repeats [(CRISPR)/CRISPR-associated protein 9 (Cas9)] genome editing. We performed ectopic expression using nucleofection and attained CRISPR-Cas9 editing via homology-dependent recombination. Targeting the E3 ubiquitin ligase x-linked inhibitor of apoptosis (xiap) and its substrate p47 led to an alteration in molecular signaling within the immune deficiency network and increased infection of the rickettsial agent Anaplasma phagocytophilum in I. scapularis ISE6 cells. Collectively, our findings complement techniques for the genetic engineering of I. scapularis ticks, which currently limit efficient and scalable molecular genetic screens in vivo.IMPORTANCEGenetic engineering in arachnids has lagged compared to insects, largely because of substantial differences in their biology. This study unveils the implementation of ectopic expression and CRISPR-Cas9 gene editing in a tick cell line. We introduced fluorescently tagged proteins in ISE6 cells and edited its genome via homology-dependent recombination. We ablated the expression of xiap and p47, two signaling molecules present in the immune deficiency (IMD) pathway of Ixodes scapularis. Impairment of the tick IMD pathway, an analogous network of the tumor necrosis factor receptor in mammals, led to enhanced infection of the rickettsial agent Anaplasma phagocytophilum. Altogether, our findings provide a critical technical resource to the scientific community to enable a deeper understanding of biological circuits in the black-legged tick I. scapularis.

Microbiome compositions of nymphal blacklegged ticks (Ixodes scapularis) infected and uninfected with Borrelia burgdorferi in Delaware

Microbiome compositions of nymphal blacklegged ticks (Ixodes scapularis) infected and uninfected with Borrelia burgdorferi in Delaware

Arthropod repellent interactions with olfactory receptors and ionotropic receptors analyzed by molecular modeling

The main insect chemoreceptors are olfactory receptors (ORs), gustatory receptors (GRs) and ionotropic receptors (IRs). The odorant binding sites of many insect ORs appear to be occluded and inaccessible from the surface of the receptor protein, based on the three-dimensional structure of OR5 from the jumping bristletail Machilis hrabei (MhraOR5) and a survey of a sample of vinegar fly (Drosophila melanogaster) OR structures obtained from artificial intellegence (A.I.) modeling. Molecular dynamics simulations revealed that the occluded site can become accessible through tunnels that transiently open and close. The present study extends this analysis to examine seventeen ORs and one GR docking with ligands that have known valence: nine that signal attraction and nine that signal aversion. All but one of the receptors displayed occluded ligand binding sites analogous to MhraOR5, and docking software predicted the known attractant and repellent ligands will bind to the occluded sites. Docking of the repellent DEET was examined, and more than half of the OR ligand sites were predicted to bind DEET, including receptors that signal aversion as well as those that signal attraction. However, DEET may not actually have access to all the attractant binding sites. The larger size and lower flexibility of repellent molecules may restrict their passage through the tunnel bottlenecks, which could act as filters to select access to the ligand binding sites. In contrast to ORs and GRs, the IR ligand binding site is in an extracellular domain known to undergo a large conformational change from an open to a closed state. A.I. models of two D. melanogaster IRs of known valence and two blacklegged tick (Ixodes scapularis) IRs having unknown ligands were computationally tested for attractant and repellent binding. The ligand-binding sites in the closed state appear inaccessible to the protein surface, so attractants and repellents must bind initially at an accessible site in the open state before triggering the conformational change. In some IRs, repellent binding sites were identified at exterior sites adjacent to the ligand-binding site. These may be allosteric sites that, when occupied by repellents, can stabilize the open state of an attractant IR, or stabilize the closed state of an IR in the absence of its activating ligand. The model of D. melanogaster IR64a suggests a possible molecular mechanism for the activation of this IR by H+. The amino acids involved in this proposed mechanism are conserved in IR64a from several Dipteran pest species and disease vectors, potentially offering a route to discovery of new repellents that act via the allosteric site.

Differential burdens of blacklegged ticks (Ixodes scapularis) on sympatric rodent hosts.

In the United States, there has been a steady increase in diagnosed cases of tick-borne diseases in people, most notably Lyme disease. The pathogen that causes Lyme disease, Borrelia burgdorferi, is transmitted by the blacklegged tick (Ixodes scapularis). Several small mammals are considered key reservoirs of this pathogen and are frequently-used hosts by blacklegged ticks. However, limited studies have evaluated between-species host use by ticks. This study compared I. scapularis burdens and tick-associated pathogen presence in wild-caught Clethrionomys gapperi (southern red-backed voles) and Peromyscus spp. (white-footed mice) in forested areas where the habitat of both species overlapped. Rodent trapping data collected over two summers showed a significant difference in the average tick burden between species. Adult Peromyscus spp. had an overall mean of 4.03 ticks per capture, while adult C. gapperi had a mean of 0.47 ticks per capture. There was a significant association between B. burgdorferi infection and host species with more Peromyscus spp. positive samples than C. gapperi (65.8% and 10.2%, respectively). This work confirms significant differences in tick-host use and pathogen presence between sympatric rodent species. It is critical to understand tick-host interactions and tick distributions to develop effective and efficient tick control methods.

Spatial variation in risk for tick-borne diseases in residential areas of Dutchess County, New York

Although human exposure to the ticks that transmit Lyme-disease bacteria is widely considered to occur around people’s homes, most studies of variation in tick abundance and infection are undertaken outside residential areas. Consequently, the patterns of variation in risk of human exposure to tick-borne infections in these human-dominated landscapes are poorly understood. Here, we report the results of four years of sampling for tick abundance, tick infection, tick encounters, and tick-borne disease reports on residential properties nested within six neighborhoods in Dutchess County, New York, USA, an area of high incidence for Lyme and other tick-borne diseases. All properties were within neighborhoods that had been randomly assigned as placebo controls in The Tick Project; hence, none were treated to reduce tick abundance during the period of investigation, providing a unique dataset of natural variation within and between neighborhoods. We estimated the abundance of host-seeking blacklegged ticks (Ixodes scapularis) in three types of habitats on residential properties–forests, lawns, and gardens. In forest and lawn habitats, some neighborhoods had consistently higher tick abundance. Properties within neighborhoods also varied consistently between years, suggesting hot spots and cold spots occurring at a small (~ 1-hectare) spatial scale. Across neighborhoods, the abundance of nymphal ticks was explained by neither the amount of forest in that neighborhood, nor by the degree of forest fragmentation. The proportion of ticks infected with three common tick-borne pathogens did not differ significantly between neighborhoods. We observed no effect of tick abundance on human encounters with ticks, nor on either human or pet cases of tick-borne diseases. However, the number of encounters between ticks and outdoor pets in a neighborhood was negatively correlated with the abundance of questing ticks in that neighborhood. Our results reinforce the need to understand how human behavior and neglected ecological factors affect variation in human encounters with ticks and cases of tick-borne disease in residential settings.

Risky business: human-related data is lacking from Lyme disease risk models.

Used as a communicative tool for risk management, risk maps provide a service to the public, conveying information that can raise risk awareness and encourage mitigation. Several studies have utilized risk maps to determine risks associated with the distribution of Borrelia burgdorferi, the causal agent of Lyme disease in North America and Europe, as this zoonotic disease can lead to severe symptoms. This literature review focused on the use of risk maps to model distributions of B. burgdorferi and its vector, the blacklegged tick (Ixodes scapularis), in North America to compare variables used to predict these spatial models. Data were compiled from the existing literature to determine which ecological, environmental, and anthropic (i.e., human focused) variables past research has considered influential to the risk level for Lyme disease. The frequency of these variables was examined and analyzed via a non-metric multidimensional scaling analysis to compare different map elements that may categorize the risk models performed. Environmental variables were found to be the most frequently used in risk spatial models, particularly temperature. It was found that there was a significantly dissimilar distribution of variables used within map elements across studies: Map Type, Map Distributions, and Map Scale. Within these map elements, few anthropic variables were considered, particularly in studies that modeled future risk, despite the objective of these models directly or indirectly focusing on public health intervention. Without including human-related factors considering these variables within risk map models, it is difficult to determine how reliable these risk maps truly are. Future researchers may be persuaded to improve disease risk models by taking this into consideration.

Resource selection by New York City deer reveals the effective interface between wildlife, zoonotic hazards and humans.

Although the role of host movement in shaping infectious disease dynamics is widely acknowledged, methodological separation between animal movement and disease ecology has prevented researchers from leveraging empirical insights from movement data to advance landscape scale understanding of infectious disease risk. To address this knowledge gap, we examine how movement behaviour and resource utilization by white-tailed deer (Odocoileus virginianus) determines blacklegged tick (Ixodes scapularis) distribution, which depend on deer for dispersal in a highly fragmented New York City borough. Multi-scale hierarchical resource selection analysis and movement modelling provide insight into how deer's movements contribute to the risk landscape for human exposure to the Lyme disease vector-I. scapularis. We find deer select highly vegetated and accessible residential properties which support blacklegged tick survival. We conclude the distribution of tick-borne disease risk results from the individual resource selection by deer across spatial scales in response to habitat fragmentation and anthropogenic disturbances.

Passive Surveillance of SARS-CoV-2 in Adult Blacklegged Ticks (Ixodes scapularis) from Northeast Pennsylvania.

Monitoring the spread of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) in wildlife is vital to public health. White-tailed deer (Odocoileus virginianus) in the United States have tested positive for SARS-CoV-2, and their interactions with blacklegged ticks (Ixodes scapularis) raise the question of whether or not these ticks also carry SARS-CoV-2. In this study, 449 blacklegged ticks from Northeast Pennsylvania were collected in the fall of 2022 and tested via RT-qPCR for the presence of SARS-CoV-2. Fourteen ticks were amplified with late quantification cycles (Cq) using primers from two nucleocapsid genes (N1 and N2) via TaqMan assays. Three of these samples were amplified on a SYBR green assay during secondary testing. However, melt curve and gel electrophoresis analysis verified negative results for these three samples. Genetic sequencing was performed on one of the three samples to look for potential cross-reactions causing the amplification observed. However, no significant match was found in the NCBI database. Although all 449 blacklegged ticks were negative for SARS-CoV-2, I. scapularis should continue to be tested for COVID-19. If blacklegged ticks test positive for COVID-19 in the future, research should focus on determining the stability of SARS-CoV-2 with the tick vector and the potential for transmission through tick bites.

Development of a low-dose fipronil deer feed: Bait-screening and range-finding to determine the optimal formulation to control blacklegged ticks (Ixodes scapularis) feeding on white-tailed deer (Odocoileus virginianus).

Lyme disease is the most prevalent vector-borne disease in the U.S., and acaricidal feeds administered to white-tailed deer (Odocoileus virginianus) have potential to disrupt blood feeding by the blacklegged tick, Ixodes scapularis. Two studies were conducted with the aim of determining an ideal formulation to deliver oral acaricides to white-tailed deer and finding the lowest fipronil dose level to effectively control I. scapularis. During formulation screening, various commercial attractants (baits) were presented to deer in large paddocks under semi-field conditions and motion-sensitive cameras were used to monitor consumption by deer and non-target species. During dose range-finding, deer were housed individually and presented a fipronil feed at one of five dose levels for 48 h (two deer untreated). At 24 h post-exposure, the deer were anesthetized, blood was drawn, and deer were manually infested with 20 I. scapularis mating pairs in feeding capsules. Tick mortality was monitored up to nine days post-attachment. Results of formulation screening indicated that loose/granular formulations were substantially more palatable to deer, relative to block formulations, and would be more effective under field conditions. Dose range-finding concluded fipronil feeds with doses of 25 ppm and higher would eliminate 100% of I. scapularis parasitizing deer. Additionally, 10 ppm fipronil feed controlled a considerable proportion of ticks, and results suggested 100% ticks could be eliminated if fipronil sulfone was present in plasma at ≥25.1 ppb. These results were paramount in developing a low dose fipronil deer feed for tick control and should provide valuable insights prior to execution of future field trials.

Operational Considerations for Using Deer-Targeted 4-Poster Tick Control Devices in a Tick-borne Disease Endemic Community.

In the northeastern United States, recommendations to prevent diseases spread by black-legged ticks ( Ixodes scapularis ) and lone star ticks ( Amblyomma americanum ) often rely on individuals to use personal protection or yard-based strategies. The 4-Poster deer treatment stations (4-Posters) suppress tick populations by treating deer hosts with acaricide, potentially offering a community-wide approach for reducing tick-borne diseases in endemic areas. The 4-Poster deployment logistics in mainland community settings are not well documented but are needed for future public health tick control efforts. As part of a public health research effort to design a population-based 4-Poster effectiveness study aimed at reducing tick-borne disease incidence, TickNET researchers partnered with the Town of Ridgefield (Connecticut) to understand the feasibility and operational logistics of deploying 4-Posters on public land within a residential community to inform future public health interventions by municipalities or vector control agencies. We deployed three 4-Posters on a municipal property from July to December 2020 and used motion-activated cameras to record wildlife activity nearby. We documented per-device operational details, costs, materials consumed, and animal activity. Operation of 4-Posters was feasible, and device challenges were easily remedied. Deer visitation and heavy nontarget animal use were documented at all devices. Unexpectedly, monthly corn consumption was not correlated with monthly deer-view days. The monthly cost per device was US $1279 or US $305 per hectare with an average 21 minutes of weekly service time. Use of 4-Posters by communities, public health agencies, or vector control programs may be a practicable addition to tick management programs in tick-borne disease endemic areas in the Northeast. Such programs should carefully consider local and state regulations, follow manufacturer and pesticide label guidelines, and include wildlife monitoring. High labor costs incurred in this project could be mitigated by training vector control agency or municipality staff to service 4-Posters.

Community-based integrated tick management programs: cost and feasibility scenarios.

Numerous studies have assessed the efficacy of environmentally based control methods to suppress populations of the blacklegged tick (Ixodes scapularis Say), but few of these estimated the cost of control. We estimated costs for a range of tick control methods (including habitat management, deer exclusion or population reduction, broadcast of acaricides, and use of host-targeted acaricides) implemented singly or in combination and applied to a model community comprising 320 residential properties and parklands. Using the high end for cost ranges, tick control based on a single method was estimated to have mean annual costs per household in the model community ranging from $132 for treating only forest ecotone with a broadcast synthetic acaricide to kill host-seeking ticks (or $404 for treating all residential forested habitat) to >$2,000 for deployment of bait boxes (SELECT TCS) across all residential tick habitat to treat rodents topically with acaricide to kill infesting ticks. Combining different sets of multiple methods in an integrated tick management program placed the annual cost between $508 and 3,192 annually per household in the model community, underscoring the disconnect between what people in Lyme disease endemic areas say they are willing to pay for tick control (not more than $100-150 annually) and the actual costs for tick control. Additional barriers to implementing community-based tick management programs within residential communities are discussed.

Acceptability of 4-poster deer treatment devices for community-wide tick control among residents of high Lyme disease incidence counties in Connecticut and New York, USA

The 4-Poster Tick Control Deer Feeder (4-poster) device applies acaricide to white-tailed deer (Odocoileus virginianus) and can reduce populations of the blacklegged tick (Ixodes scapularis), which transmits the agents of Lyme disease, anaplasmosis, babesiosis, and Powassan virus disease in the Northeastern United States. While 4-poster devices have the potential to provide community-wide management of blacklegged ticks in Lyme disease endemic areas, no recent study has assessed their acceptability among residents. We conducted a survey of residents from 16 counties with high annual average Lyme disease incidence (≥ 10 cases per 100,000 persons between 2013 and 2017) in Connecticut and New York to understand perceptions and experiences related to tickborne diseases, support or concerns for placement of 4-poster devices in their community, and opinions on which entities should be responsible for tick control on private properties. Overall, 37% of 1652 respondents (5.5% response rate) would support placement of a 4-poster device on their own property, 71% would support placement on other private land in their community, and 90% would support placement on public land. Respondents who were male, rented their property, resided on larger properties, or were very or extremely concerned about encountering ticks on their property were each more likely to support placement of 4-poster devices on their own property. The primary reason for not supporting placement of a 4-poster device on one's own property was the need for weekly service visits from pest control professionals, whereas the top reason for not supporting placement on other land (private or public) was safety concerns. Most respondents (61%) felt property owners should be responsible for tick control on private properties. Communities considering 4-poster devices as part of a tick management strategy should consider targeting owners of larger properties and placing devices on public lands.

Tick Densities and Infection Prevalence on Coastal Islands in Massachusetts, USA: Establishing a Baseline.

Tick-borne diseases and a tick-induced red meat allergy have become increasingly common in the northeastern USA and elsewhere. At the scale of local communities, few studies have documented tick densities or infection levels to characterize current conditions and provide a baseline for further monitoring. Using the town of Nantucket, MA, as a case study, we recorded tick densities by drag sampling along hiking trails in nature preserves on two islands. Nymphal blacklegged ticks (Ixodes scapularis Say) were most abundant at shadier sites and least common in grasslands and scrub oak thickets (Quercus ilicifolia). Lone star ticks (Amblyomma americanum L.) were common on Tuckernuck Island and rare on Nantucket Island, while both tick species were more numerous in 2021 compared to 2020 and 2022. We tested for pathogens in blacklegged nymphs at five sites over two years. In 2020 and 2021, infection levels among the four Nantucket Island sites averaged 10% vs. 19% for Borrelia burgdorferi, 11% vs. 15% for Babesia microti, and 17% (both years) for Anaplasma phagocytophilum, while corresponding levels were significantly greater on Tuckernuck in 2021. Our site-specific, quantitative approach represents a practical example of how potential exposure to tick-borne diseases can be monitored on a local scale.

The principal eigenvalue for partially degenerate and periodic reaction-diffusion systems with time delay

In this paper, we first establish the theory of principal eigenvalues for a large class of partially degenerate, linear and periodic parabolic cooperative systems with time delay. Then we apply these theoretical results to study the global dynamics of a blacklegged tick Ixodes scapularis population model. To present a threshold-type result in terms of basic reproduction ratio R0 for such a model, we also extend the earlier theory of R0 to abstract functional differential equations with time-delayed internal transition.

Deer management generally reduces densities of nymphal Ixodes scapularis, but not prevalence of infection with Borrelia burgdorferi sensu stricto

Human Lyme disease–primarily caused by the bacterium Borrelia burgdorferi sensu stricto (s.s.) in North America–is the most common vector-borne disease in the United States. Research on risk mitigation strategies during the last three decades has emphasized methods to reduce densities of the primary vector in eastern North America, the blacklegged tick (Ixodes scapularis). Controlling white-tailed deer populations has been considered a potential method for reducing tick densities, as white-tailed deer are important hosts for blacklegged tick reproduction. However, the feasibility and efficacy of white-tailed deer management to impact acarological risk of encountering infected ticks (namely, density of host-seeking infected nymphs; DIN) is unclear. We investigated the effect of white-tailed deer density and management on the density of host-seeking nymphs and B. burgdorferi s.s. infection prevalence using surveillance data from eight national parks and park regions in the eastern United States from 2014–2022. We found that deer density was significantly positively correlated with the density of nymphs (nymph density increased by 49% with a 1 standard deviation increase in deer density) but was not strongly correlated with the prevalence of B. burgdorferi s.s. infection in nymphal ticks. Further, while white-tailed deer reduction efforts were followed by a decrease in the density of I. scapularis nymphs in parks, deer removal had variable effects on B. burgdorferi s.s. infection prevalence, with some parks experiencing slight declines and others slight increases in prevalence. Our findings suggest that managing white-tailed deer densities alone may not be effective in reducing DIN in all situations but may be a useful tool when implemented in integrated management regimes.

Multiomics Reveals Symbionts, Pathogens, and Tissue-Specific Microbiome of Blacklegged Ticks (Ixodes scapularis) from a Lyme Disease Hot Spot in Southeastern Ontario, Canada.

Ticks in the family Ixodidae are important vectors of zoonoses, including Lyme disease (LD), which is caused by spirochete bacteria from the Borreliella (Borrelia) burgdorferi sensu lato complex. The blacklegged tick (Ixodes scapularis) continues to expand across Canada, creating hot spots of elevated LD risk at the leading edge of its expanding range. Current efforts to understand the risk of pathogen transmission associated with I. scapularis in Canada focus primarily on targeted screens, while natural variation in the tick microbiome remains poorly understood. Using multiomics consisting of 16S metabarcoding and ribosome-depleted, whole-shotgun RNA transcriptome sequencing, we examined the microbial communities associated with adult I. scapularis (n = 32), sampled from four tissue types (whole tick, salivary glands, midgut, and viscera) and three geographical locations within a LD hot spot near Kingston, Ontario, Canada. The communities consisted of both endosymbiotic and known or potentially pathogenic microbes, including RNA viruses, bacteria, and a Babesia sp. intracellular parasite. We show that β-diversity is significantly higher between the bacterial communities of individual tick salivary glands and midguts than that of whole ticks. Linear discriminant analysis effect size (LEfSe) determined that the three potentially pathogenic bacteria detected by V4 16S rRNA sequencing also differed among dissected tissues only, including a Borrelia strain from the B. burgdorferi sensu lato complex, Borrelia miyamotoi, and Anaplasma phagocytophilum. Importantly, we find coinfection of I. scapularis by multiple microbes, in contrast to diagnostic protocols for LD, which typically focus on infection from a single pathogen of interest (B. burgdorferi sensu stricto). IMPORTANCE As a vector of human health concern, blacklegged ticks (Ixodes scapularis) transmit pathogens that cause tick-borne diseases (TBDs), including Lyme disease (LD). Several hot spots of elevated LD risk have emerged across Canada as I. scapularis expands its range. Focusing on a hot spot in southeastern Ontario, we used high-throughput sequencing to characterize the microbiome of whole ticks and dissected salivary glands and midguts. Compared with whole ticks, salivary glands and midguts were more diverse and associated with distinct bacterial communities that are less dominated by Rickettsia endosymbiont bacteria and are enriched for pathogenic bacteria, including a B. burgdorferi sensu lato-associated Borrelia sp., Borrelia miyamotoi, and Anaplasma phagocytophilum. We also found evidence of coinfection of I. scapularis by multiple pathogens. Overall, our study highlights the challenges and opportunities associated with the surveillance of the microbiome of I. scapularis for pathogen detection using metabarcoding and metatranscriptome approaches.

Ticks harbor and excrete chronic wasting disease prions

Chronic wasting disease (CWD) is a fatal neurodegenerative disease caused by infectious prions (PrPCWD) affecting cervids. Circulating PrPCWD in blood may pose a risk for indirect transmission by way of hematophagous ectoparasites acting as mechanical vectors. Cervids can carry high tick infestations and exhibit allogrooming, a common tick defense strategy between conspecifics. Ingestion of ticks during allogrooming may expose naïve animals to CWD, if ticks harbor PrPCWD. This study investigates whether ticks can harbor transmission-relevant quantities of PrPCWD by combining experimental tick feeding trials and evaluation of ticks from free-ranging white-tailed deer (Odocoileus virginianus). Using the real-time quaking-induced conversion (RT-QuIC) assay, we show that black-legged ticks (Ixodes scapularis) fed PrPCWD-spiked blood using artificial membranes ingest and excrete PrPCWD. Combining results of RT-QuIC and protein misfolding cyclic amplification, we detected seeding activity from 6 of 15 (40%) pooled tick samples collected from wild CWD-infected white-tailed deer. Seeding activities in ticks were analogous to 10–1000 ng of CWD-positive retropharyngeal lymph node collected from deer upon which they were feeding. Estimates revealed a median infectious dose range of 0.3–42.4 per tick, suggesting that ticks can take up transmission-relevant amounts of PrPCWD and may pose a CWD risk to cervids.