Plague is a rare, potentially fatal flea-borne zoonosis endemic in the western United States. A previous model described interannual variation in human cases based on temperature and lagged precipitation. We recreated this model in northeastern Arizona (1960-1997) to evaluate its capacity to predict recent cases (1998-2022). In recreating the original model, we found that future instead of concurrent temperature had inadvertently been used for the presented fit. Prediction from our revised models with lagged precipitation and temporally plausible temperature relationships aligned with low observed cases in 1998-2022. Elevated precipitation associated with high cases in historical data (>6 inches combined precipitation over two previous springs) was only observed once in the last quarter century, so we could not assess if these conditions were reliably associated with elevated (four or more) human plague cases. Observed weather conditions were similar to those previously associated with low (fewer than or equal to two) case counts, suggesting "baseline" conditions in the last quarter century.

Knowledge of seasonal activity patterns of human-biting life stages of tick species serving as vectors of human disease agents provides basic information on when during the year humans are most at risk for tick bites and tick-borne diseases. Although there is a wealth of published information on seasonal activity patterns of Ixodes scapularis and Ixodes pacificus in the United States, a critical review of the literature for these important tick vectors is lacking. The aims of this paper were to: (i) review what is known about the seasonal activity patterns of I. scapularis and I. pacificus in different parts of their geographic ranges in the US, (ii) provide a synthesis of the main findings, and (iii) outline key knowledge gaps and methodological pitfalls that limit our understanding of variability in seasonal activity patterns. Based on ticks collected while questing or from wild animals, the seasonal activity patterns were found to be similar for I. pacificus in the Far West and I. scapularis in the Southeast, with synchronous activity of larvae and nymphs, peaking in spring (April to June) in the Far West and from spring to early summer (April to July) in the Southeast, and continuous activity of adults from fall through winter and spring with peak activity from fall through winter (November/December to March). In the colder climates of the Upper Midwest and Northeast, I. scapularis adults have a bimodal seasonal pattern, with activity peaks in fall (October to November) and spring (April to May). The seasonal activity patterns for immatures differ between the Upper Midwest, synchronous for larvae and nymphs with peak activity in spring and summer (May to August), and the Northeast, where the peak activity of nymphs in spring and early summer (May to July) precedes that of larvae in summer (July to September). Seasonality of human tick encounters also is influenced by changes over the year in the level of outdoor activities in tick habitat. Studies on the seasonality of ticks infesting humans have primarily focused on the coastal Northeast and the Pacific Coast states, with fewer studies in the Southeast, inland parts of the Northeast, and the Upper Midwest. Discrepancies between seasonal patterns for peak tick questing activity and peak human infestation appear to occur primarily for the adult stages of I. scapularis and I. pacificus. Study design and data presentation limitations of the published literature are discussed. Scarcity of data for seasonal activity patterns of I. pacificus outside of California and for I. scapularis from parts of the Southeast, Northeast, and Upper Midwest is a key knowledge gap. In addition to informing the public of when during the year the risk for tick bites is greatest, high-quality studies describing current seasonal activity patterns also will generate the data needed for robust model-based projections of future climate-driven change in the seasonal activity patterns and provide the baseline needed to empirically determine in the future if the projections were accurate.

A previous laboratory study using Haemaphysalis longicornis Neumann (Acari: Ixodidae) ticks of North American origin showed that larvae could acquire the Lyme disease spirochete, Borrelia burgdorferi sensu stricto (s.s.) (Spirochaetales: Spirochaetaceae) while feeding to completion on infected mice. However, the infection was lost during the molt to the nymphal stage. Nonetheless, questing H. longicornis nymphs and adults collected by drag sampling in the northeastern United States have been reported infected with B. burgdorferi s.s. DNA; occasionally these ticks appeared to be partially engorged. This raises the question of whether H. longicornis ticks can (i) acquire B. burgdorferi s.s. during an interrupted, partial blood meal on an infected host and (ii) transmit spirochetes while completing the blood meal on a second host. In this laboratory study, we demonstrated that H. longicornis nymphs could acquire B. burgdorferi s.s. from infected Mus musculus mice during a partial blood meal. Borrelia burgdorferi s.s. was detected by a multiplex polymerase chain reaction amplicon sequencing assay in 2 of 32 (6.3%) nymphs allowed to remain attached to infected mice for 48 h but, paradoxically, not in any of 25 nymphs that remained attached to infected mice for 72 h. Unfortunately, due to the low percentage of infected nymphs, we were not able to examine if such partially fed, infected nymphs were able to transmit B. burgdorferi s.s. while completing their blood meal on a second, naïve host.

In response to notable increases in tick-associated illnesses in the United States, recent public health policies encouraged multi-sector collaborative approaches to preventing vector-borne diseases. Primary prevention strategies focus on educating the public about risks for tick-borne diseases and encouraging adoption of personal protection strategies. Accurate descriptions of when and where people are at risk for tick-borne diseases aid in the optimization of prevention messaging. Tick and tick-borne pathogen data can be used to fill gaps in epidemiological surveillance. However, the utility of acarological data is limited by their completeness. National maps showing the distribution of medically important tick species and the pathogens they carry are often incomplete or non-existent. Recent policies encourage accelerated efforts to monitor changes in the distribution and abundance of medically important ticks and the presence and prevalence of human pathogens that they carry, and to provide actionable, evidence-based information to the public, health care providers and public health policy makers. In 2018, the Centers for Disease Control and Prevention initiated a national tick surveillance program focused on Ixodes ticks. The national program coordinated and expanded upon existing efforts led by public health departments and academic institutions. Here, we describe experiences of state public health departments engaged in Ixodes tick surveillance, including information on why they initiated Ixodes surveillance programs, programmatic objectives, and strategies for maintaining tick surveillance programs. We share experiences and challenges in interpreting or communicating tick surveillance data to stakeholders and explore how the acarological data are used to complement epidemiological data.

BACKGROUND: Ixodes scapularis and Ixodes pacificus are important vectors of multiple pathogens in the United States. However, their role in transmission of Bartonella spp., which are commonly reported in rodents and fleas, has been debated. Our previous investigation on Bartonella spp. in host-seeking I. scapularis and I. pacificus showed Bartonella spp. were absent in the ticks, suggesting the two species are unlikely to contribute to Bartonella transmission. It is unclear whether the absence of Bartonella spp. in the host-seeking ticks was attributable to ticks not being exposed to Bartonella in nature or being exposed but unable to acquire or transstadially transmit the bacterium. To assess the likelihood of exposure and acquisition, we tested Ixodes spp. ticks collected from rodents for Bartonella infections. METHODS: Blood-fed I. scapularis ticks (n = 792; consisting of 645 larvae and 147 nymphs), I. pacificus ticks (n = 45, all larvae), and Ixodes angustus ticks (n = 16, consisting of 11 larvae and 5 nymphs) collected from rodents from Minnesota and Washington were tested for Bartonella spp. using a quadruplex polymerase chain reaction (PCR) amplicon next-generation sequencing approach that targets Bartonella-specific fragments on gltA, ssrA, rpoB, and groEL. In parallel, rodents and fleas collected from the same field studies were investigated to compare the differences of Bartonella distribution among the ticks, fleas, and rodents. RESULTS: Bartonella spp. were commonly detected in rodents and fleas, with prevalence of 25.6% in rodents and 36.8% in fleas from Minnesota; 27.9% in rodents and 45.2% in fleas from Washington. Of all tested ticks, Bartonella DNA was detected by gltA in only one larval I. scapularis tick from Minnesota. CONCLUSIONS: The high prevalence of Bartonella spp. in rodents and fleas coupled with extremely low prevalence of Bartonella spp. in blood-fed ticks suggests that although Ixodes ticks commonly encounter Bartonella in rodents, they rarely acquire the infection through blood feeding. Notably, ticks were at various stages of feeding on rodents when they were collected. Laboratory transmission studies are needed to assess acquisition rates in fully blood-fed ticks and to assess transstadial transmission efficiency if ticks acquire Bartonella infections from feeding to repletion.

We report the genomic sequence of the hard tick relapsing fever spirochete Borrelia miyamotoi strain MN18-0001. B. miyamotoi causes human illness and is geographically widespread in Ixodes spp. (Acari: Ixodidae) ticks. This is a chromosome- and plasmid-resolved genome assembly of an Am-East-2 strain type isolate from the midwestern United States.

BACKGROUND: The vast majority of vector-borne diseases in the USA are associated with mosquitoes or ticks. Mosquito control is often conducted as part of community programs run by publicly-funded entities. By contrast, tick control focuses primarily on individual residential properties and is implemented predominantly by homeowners and the private pest control firms they contract. We surveyed publicly-funded vector control programs (VCPs), presumed to focus mainly on mosquitoes, to determine what tick-related services they currently offer, and their interest in and capacity to expand existing services or provide new ones. METHODS: We distributed a survey to VCPs in the Northeast, Upper Midwest and Pacific Coast states of the USA, where humans are at risk for bites by tick vectors (Ixodes scapularis or Ixodes pacificus) of agents causing Lyme disease and other tick-borne diseases. The data we report are based on responses from 118 VCPs engaged in vector control and with at least some activities focused on ticks. RESULTS: Despite our survey targeting geographic regions where ticks and tick-borne diseases are persistent and increasing public health concerns, only 11% (12/114) of VCPs reported they took direct action to suppress ticks questing in the environment. The most common tick-related activities conducted by the VCPs were tick bite prevention education for the public (70%; 75/107 VCPs) and tick surveillance (48%; 56/116). When asked which services they would most likely include as part of a comprehensive tick management program, tick bite prevention education (90%; 96/107), tick surveillance (89%; 95/107) and tick suppression guidance for the public (74%; 79/107) were the most common services selected. Most VCPs were also willing to consider engaging in activities to suppress ticks on public lands (68%; 73/107), but few were willing to consider suppressing ticks on privately owned land such as residential properties (15%; 16/107). Across all potential tick-related services, funding was reported as the biggest obstacle to program expansion or development, followed by personnel. CONCLUSIONS: Considering the hesitancy of VCPs to provide tick suppression services on private properties and the high risk for tick bites in peridomestic settings, suppression of ticks on residential properties by private pest control operators will likely play an important role in the tick suppression landscape in the USA for the foreseeable future. Nevertheless, VCPs can assist in this effort by providing locally relevant guidelines to homeowners and private pest control firms regarding best practices for residential tick suppression efforts and associated efficacy evaluations. Publicly-funded VCPs are also well positioned to educate the public on personal tick bite prevention measures and to collect tick surveillance data that provide information on the risk of human encounters with ticks within their jurisdictions.

Human anaplasmosis cases, caused by Anaplasma phagocytophilum, are increasing in the United States. This trend is explained, in part, by expansion in the geographic range of the primary vector, Ixodes scapularis. Multiple variants of A. phagocytophilum have been identified in field collected ticks, but only a single variant (human active, or "Ap-ha," variant) has been shown to be pathogenic in humans. Until recently, laboratory methods used to differentiate variants were cumbersome and seldomly used in large scale assessments of the pathogen's geographic distribution. As a result, many surveys reported A. phagocytophilum without segregating variants. Lack of discrimination among A. phagocytophilum variants could lead to overestimation of anaplasmosis risk to humans. Next Generation Sequencing (NGS) assays were recently developed to efficiently detect multiple Ixodes scapularis-borne human pathogens including Ap-ha. In this study, we utilized NGS to detect and differentiate A. phagocytophilum variants (Ap-ha vs. non ha) in host-seeking I. scapularis nymphs and adults collected across 23 states in the eastern United States from 2012 to 2023 as part of national tick surveillance efforts and research studies. Many of the included ticks were tested previously using a TaqMan PCR assay that could detect A. phagocytophilum but could not differentiate variants. We retested A. phagocytophilum infected ticks with NGS to differentiate variants. Anaplasma phagocytophilum (any variant) was identified in 165 (35 %) of 471 counties from which ticks were tested, whereas Ap-ha was detected in 70 (15 %) of 469 counties where variants were differentiated. Both variants were identified in 32 % (n = 40) of 126 counties with either variant detected. Among states where A. phagocytophilum (any variant) was detected, prevalence ranged from 2 % to 19 % in unfed adults and from 0.2 % to 7.8 % in unfed nymphs; prevalence of Ap-ha variant ranged from 0.0 % to 16 % in adults, and 0.0 % to 4.6 % in nymphs.

We conducted surveys of New Jersey mosquito control and public health agencies to determine their willingness and ability to expand or create and maintain publicly funded tick and tick-borne disease (T/TBD) management programs. Nearly all (86%) of 21 county mosquito control agencies (MCAs) completed the survey, while only 25% of the 102 health departments (HDs) responded, probably reflecting traditional agency responsibilities. Although few of either group had formal programs, many were engaged in T/TBD-related activities. Many MCAs rated their ability to assume T/TBD responsibilities as high or moderate, while most HDs rated their capabilities as low. With the exceptions of lack of sustainable funding and possible legal constraints, the groups differed regarding perceived barriers to program creation and maintenance. Both groups envisioned comprehensive programs emphasizing public education, but program priorities differed between the groups. MCAs were willing to include most program activities, while HDs felt that some activities should be the responsibility of other agencies. MCAs were generally more familiar than HDs with tick control methods and while both groups would include control in a comprehensive program, both would limit control to public lands. Estimated program costs varied widely, probably reflecting responding agency size and complexity of envisioned programs. These results in a state with a system of existing agencies staffed by highly competent professionals suggest that more than simply additional funding (e.g., established guidelines for tick control and surveillance) is needed to create a network of practice necessary to address the growing incidence of TBD.

The burden of tick-borne diseases continues to increase in the United States. Tick surveillance has been implemented to monitor changes in the distribution and prevalence of human disease-causing pathogens in ticks that frequently bite humans. Such efforts require accurate identification of ticks to species and highly sensitive and specific assays that can detect and differentiate pathogens from genetically similar microbes in ticks that have not been demonstrated to be pathogenic in humans. We describe a modification to a next generation sequencing pathogen detection assay that includes a target that accurately identifies Ixodes ticks to species. We show that the replacement of internal control primers used to ensure assay performance with primers that also act as an internal control and can additionally differentiate tick species, retains high sensitivity and specificity, improves efficiency, and reduces costs by eliminating the need to run separate assays to screen for pathogens and for tick identification.

[This corrects the article DOI: 10.3389/fmicb.2023.1243471.].

Ixodes pacificus (the western blacklegged tick) occurs in the far western United States (US), where it commonly bites humans. This tick was not considered a species of medical concern until it was implicated in the 1980s as a vector of Lyme disease spirochetes. Later, it was discovered to also be the primary vector to humans in the far western US of agents causing anaplasmosis and hard tick relapsing fever. The core distribution of I. pacificus in the US includes California, western Oregon, and western Washington, with outlier populations reported in Utah and Arizona. In this review, we provide a history of the documented occurrence of I. pacificus in the US from the 1890s to present, and discuss associations of its geographic range with landscape, hosts, and climate. In contrast to Ixodes scapularis (the blacklegged tick) in the eastern US, there is no evidence for a dramatic change in the geographic distribution of I. pacificus over the last half-century. Field surveys in the 1930s and 1940s documented I. pacificus along the Pacific Coast from southern California to northern Washington, in the Sierra Nevada foothills, and in western Utah. County level collection records often included both immatures and adults of I. pacificus, recovered by drag sampling or from humans, domestic animals, and wildlife. The estimated geographic distribution presented for I. pacificus in 1945 by Bishopp and Trembley is similar to that presented in 2022 by the Centers for Disease Control and Prevention. There is no clear evidence of range expansion for I. pacificus, separate from tick records in new areas that could have resulted from newly initiated or intensified surveillance efforts. Moreover, there is no evidence from long-term studies that the density of questing I. pacificus ticks has increased over time in specific areas. It therefore is not surprising that the incidence of Lyme disease has remained stable in the Pacific Coast states from the early 1990s, when it became a notifiable condition, to present. We note that deforestation and deer depredation were less severe in the far western US during the 1800s and early 1900s compared to the eastern US. This likely contributed to I. pacificus maintaining stable, widespread populations across its geographic range in the far western US in the early 1900s, while I. scapularis during the same time period appears to have been restricted to a small number of geographically isolated refugia sites within its present range in the eastern US. The impact that a warming climate may have had on the geographic distribution and local abundance of I. pacificus in recent decades remains unclear.

BACKGROUND: The majority of vector-borne disease cases in the USA are caused by pathogens spread by ticks, most commonly the blacklegged tick, Ixodes scapularis. Personal protection against tick bites, including use of repellents, is the primary defense against tick-borne diseases. Tick repellents registered by the Environmental Protection Agency (EPA) are well documented to be safe as well as effective against ticks. Another group of tick repellent products, 25(b) exempt or minimum risk products, use alternative, mostly botanically derived, active ingredients. These are considered to pose minimal risk to human health and therefore are exempt from EPA registration; efficacy testing is not mandated for these products. METHODS: We used a finger bioassay to evaluate the repellency against I. scapularis nymphs for 11 formulated 25(b) exempt products together with two positive control DEET-based EPA registered products. Repellency was assessed hourly from 0.5 to 6.5 h after product application. RESULTS: The DEET-based products showed ≥ 97% repellency for all examined timepoints. By contrast, an average of 63% of ticks were repelled in the first 1.5 h after application across the 11 25(b) exempt products, and the average fell to 3% repelled between 2.5 and 6.5 h. Ten of the 11 25(b) exempt products showed statistically similar efficacy to DEET-based products at 30 min after application (repellency of 79-97%). However, only four 25(b) exempt products maintained a level of repellency similar to DEET-based products (> 72%) at the 1.5-h mark, and none of these products were effective in repelling ticks at the timepoints from 2.5 to 6.5 h after application. CONCLUSIONS: Neither the claims on the labels nor specific active ingredients and their concentrations appeared to predict the duration of efficacy we observed for the 25(b) exempt products. These products are not registered with the EPA, so the methods used to determine the application guidelines on their labels are unclear. Consumers should be aware that both the level of efficacy and the duration of repellency may differ among unregulated 25(b) exempt repellent products labeled for use against ticks. We encourage more research on these products and the 25(b) exempt active ingredients they contain to help determine and improve their efficacy as repellents under different conditions.

The majority of vector-borne disease cases reported annually in the United States are caused by pathogens spread by the blacklegged tick, Ixodes scapularis. The number and geographic distribution of cases have increased as the geographic range and abundance of the tick have expanded in recent decades. A large proportion of Lyme disease and other I. scapularis-borne diseases are associated with nymphal tick bites; likelihood of such bites generally increases with increasing nymphal densities. National tick surveillance was initiated in 2018 to track changes in the distribution and abundance of medically important ticks at the county spatial scale throughout the United States. Tick surveillance records, including historical data collected prior to the initiation of the national program, are collated in the ArboNET Tick Module database. Through exploration of ArboNET Tick Module data, we found that efforts to quantify the density of host-seeking I. scapularis nymphs (DON) were unevenly distributed among geographic regions with the greatest proportion of counties sampled in the Northeast and Upper Midwest. Submissions covering tick collections from 2004 through 2022 revealed extensive variation in DON estimates at collection site, county, state, and regional spatial scales. Throughout the entire study period, county DON estimates ranged from 0.0 to 488.5 nymphs/1,000 m(2). Although substantial variation was recorded within regions, DON estimates were greatest in the Northeast, Upper Midwest, and northern states within the Southeast regions (Virginia and North Carolina); densities were intermediate in the Ohio Valley and very low in the South and Northern Rockies and Plains regions. The proportion of counties classified as moderate or high DON was greater in the Northeast, Ohio Valley, and Southeast regions during the 2004 through 2017 time period (prior to initiation of the national tick surveillance program) compared to 2018 through 2022; DON estimates remained similarly low between these time periods in the South and the Northern Rockies and Plains regions. Despite the limitations described herein, the ArboNET Tick Module provides useful data for tracking changes in acarological risk across multiple geographic scales and long periods of time.

Human-biting ticks threaten public health in the United States. Registration by the Environmental Protection Agency of products to kill host-seeking ticks or repel ticks contacting humans is indicative of their safety and effectiveness. Unregulated minimum risk products, exempt from Environmental Protection Agency registration and often based on botanical oils, are proliferating in the marketplace, but there is concern about their effectiveness to kill and repel ticks. Evaluations of such products are limited in the published literature. A review showed considerable variability among minimum risk products to kill host-seeking blacklegged ticks, with effectiveness similar to chemical pesticide products for some minimum risk products but minimal impact on the ticks for other products. Evaluations of minimum risk tick repellents have typically focused on individual active ingredients rather than formulated products, which often combine multiple active ingredients. Consumers should be aware that effectiveness to kill and repel ticks can differ among unregulated minimum risk products.

The genus Bartonella includes a group of species that are associated with a wide range of mammalian species, including human. It is challenging to detect all Bartonella species using a single molecular target due to its high genetic diversity. To solve this issue, we developed a quadruplex PCR amplicon sequencing assay using next-generation sequencing (NGS) technology for the detection and differentiation of Bartonella species. Our objective was to obtain the specific sequences of a minimum of two of the four target genes as confirmation of the identity of a particular Bartonella species using the assay. Four pairs of primers targeting specific regions on gltA, groEL, rpoB, and ssrA were evaluated for their capability of differentiating Bartonella species individually and collectively by performing singular PCR amplicon sequencing and quadruplex PCR amplicon sequencing. Using the quadruplex PCR amplicon sequencing, 24 Bartonella reference species were tested, all of which were successfully differentiated by at least two targets. Bartonella species were accurately identified from the artificially mixed DNA templates developed to simulate coinfections. The limit of detection was determined to be 1 fg based on testing a series of 10-fold dilutions of DNA from the Bartonella species. Testing of high DNA concentrations of 19 non-Bartonella species showed high specificity with none of the non-Bartonella species misclassified as Bartonella. Finally, the assay was evaluated by testing DNA extracts from field-collected body lice (Pediculus humanus humanus) and Norway rats (Rattus norvegicus): Bartonella quintana was detected and confirmed by three targets in the lice and Bartonella tribocorum was detected and confirmed by two targets in the rats. These results demonstrated that Bartonella species could be accurately and rapidly detected and differentiated into different tissue types using the quadruplex sequencing assay.

Ixodes scapularis (the blacklegged tick) is widely distributed in forested areas across the eastern United States. The public health impact of I. scapularis is greatest in the north, where nymphal stage ticks commonly bite humans and serve as primary vectors for multiple human pathogens. There were dramatic increases in the tick's distribution and abundance over the last half-century in the northern part of the eastern US, and climate warming is commonly mentioned as a primary driver for these changes. In this review, we summarize the evidence for the observed spread and proliferation of I. scapularis being driven by climate warming. Although laboratory and small-scale field studies have provided insights into how temperature and humidity impact survival and reproduction of I. scapularis, using these associations to predict broad-scale distribution and abundance patterns is more challenging. Numerous efforts have been undertaken to model the distribution and abundance of I. scapularis at state, regional, and global scales based on climate and landscape variables, but outcomes have been ambiguous. Across the models, the functional relationships between seasonal or annual measures of heat, cold, precipitation, or humidity and tick presence or abundance were inconsistent. The contribution of climate relative to landscape variables was poorly defined. Over the last half-century, climate warming occurred in parallel with spread and population increase of the white-tailed deer, the most important reproductive host for I. scapularis adults, in the northern part of the eastern US. There is strong evidence for white-tailed deer playing a key role to facilitate spread and proliferation of I. scapularis in the US over the last century. However, due to a lack of spatially and temporally congruent data, climate, landscape, and host variables are rarely included in the same models, thus limiting the ability to evaluate their relative contributions or interactions in defining the geographic range and abundance patterns of ticks. We conclude that the role of climate change as a key driver for geographic expansion and population increase of I. scapularis in the northern part of the eastern US over the last half-century remains uncertain.

Lyme disease is the most commonly reported vector-borne disease in the United States and is transmitted by Ixodes scapularis in the eastern US and I. pacificus in the west. The causative agents, Borrelia burgdorferi sensu stricto (Bbss) and B. mayonii belong to the B. burgdorferi sensu lato (Bbsl) species complex. An additional eight species of Bbsl have been identified in Ixodes species ticks in the US, but their geographic distribution, vector associations, human encounter rates and pathogenicity in humans are poorly defined. To better understand the geographic distribution and vector associations of Bbsl spirochetes in frequent and infrequent human-biting Ixodes species ticks in the US, we previously screened 29,517 host-seeking I. scapularis or I. pacificus ticks and 692 ticks belonging to eight other Ixodes species for Borrelia spirochetes using a previously described tick testing algorithm that utilizes a combination of real-time PCR and Sanger sequencing for Borrelia species identification. The assay was designed to detect known human pathogens spread by Ixodes species ticks, but it was not optimized to detect Bbsl co-infections. To determine if such co-infections were overlooked particularly in ticks infected with Bbss, we retested and analyzed a subsample of 845 Borrelia infected ticks using a next generation sequencing multiplex PCR amplicon sequencing (MPAS) assay that can identify Borrelia species and Bbsl co-infections. The assay also includes targets that can molecularly confirm identifications of Ixodes species ticks to better inform pathogen-vector associations. We show that Bbss is the most prevalent species in I. scapularis and I. pacificus; other Bbsl species were rarely detected in I. scapularis and the only Bbsl co-infections identified in I. scapularis were with Bbss and B. mayonii. We detected B. andersonii in I. dentatus in the Mid-Atlantic and Upper Midwest regions, B. kurtenbachii in I. scapularis in the Upper Midwest, B. bissettiae in I. pacificus and I. spinipalpis in the Northwest, and B. carolinensis in I. affinis in the Mid-Atlantic and Southeast, and B. lanei in I. spinipalpis in the Northwest. Twelve of 62 (19.4%) Borrelia-infected I. affinis from the Mid-Atlantic region were co-infected with Bbss and B. carolinensis. Our data support the notion that Bbsl species are maintained in largely independent enzootic cycles, with occasional spill-over resulting in multiple Bbsl species detected in Ixodes species ticks.

The majority of vector-borne disease cases reported in the United States (U.S.) are caused by pathogens spread by the blacklegged tick, Ixodes scapularis. In recent decades, the geographic ranges of the tick and its associated human pathogens have expanded, putting an increasing number of communities at risk for tick-borne infections. In 2018, the U.S. Centers for Disease Control and Prevention (CDC) initiated a national tick surveillance program to monitor changes in the distribution and abundance of ticks and the presence and prevalence of human pathogens in them. We assessed the geographical representativeness of prevalence data submitted to CDC as part of the national tick surveillance effort. We describe county, state, and regional variation in the prevalence of five human pathogens (Borrelia burgdorferi sensu stricto (s.s.), Borrelia mayonii, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia microti) in host-seeking I. scapularis and I. pacificus nymphs and adults. Although I. scapularis and I. pacificus are widely distributed in the eastern and western U.S., respectively, pathogen prevalence was estimated predominantly in ticks collected in the Northeast, Ohio Valley, and Upper Midwest regions, where human Lyme disease cases are most commonly reported. Within these regions, we found that state and regional estimates of pathogen prevalence generally reached predictable and stable levels, but variation in prevalence estimates at the sub-state level was considerable. Borrelia burgdorferi s.s. was the most prevalent and widespread pathogen detected. Borrelia miyamotoi and A. phagocytophilum shared a similarly broad geographic range, but were consistently detected at much lower prevalence compared with B. burgdorferi s.s. Babesia microti was detected at similar prevalence to A. phagocytophilum, where both pathogens co-occurred, but was reported over a much more limited geographic range compared with A. phagocytophilum or B. burgdorferi s.s. Borrelia mayonii was identified at very low prevalence with a focal distribution within the Upper Midwest. National assessments of risk for tick-borne diseases need to be improved through collection and testing of ticks in currently under-represented regions, including the West, South, Southeast, and eastern Plains states.

Tick-borne diseases continue to threaten human health across the United States. Both active and passive tick surveillance can complement human case surveillance, providing spatio-temporal information on when and where humans are at risk for encounters with ticks and tick-borne pathogens. However, little work has been done to assess the concordance of the acarological risk metrics from each surveillance method. We used data on Ixodes scapularis and its associated human pathogens from Connecticut (2019-2021) collected through active collections (drag sampling) or passive submissions from the public to compare county estimates of tick and pathogen presence, infection prevalence, and tick abundance by life stage. Between the surveillance strategies, we found complete agreement in estimates of tick and pathogen presence, high concordance in infection prevalence estimates for Anaplasma phagocytophilum, Borrelia burgdorferi sensu stricto, and Babesia microti, but no consistent relationships between actively and passively derived estimates of tick abundance or abundance of infected ticks by life stage. We also compared nymphal metrics (i.e., pathogen prevalence in nymphs, nymphal abundance, and abundance of infected nymphs) with reported incidence of Lyme disease, anaplasmosis, and babesiosis, but did not find any consistent relationships with any of these metrics. The small spatial and temporal scale for which we had consistently collected active and passive data limited our ability to find significant relationships. Findings are likely to differ if examined across a broader spatial or temporal coverage with greater variation in acarological and epidemiological outcomes. Our results indicate similar outcomes between some actively and passively derived tick surveillance metrics (tick and pathogen presence, pathogen prevalence), but comparisons were variable for abundance estimates.

BACKGROUND: Bartonella spp. infect a variety of vertebrates throughout the world, with generally high prevalence. Several Bartonella spp. are known to cause diverse clinical manifestations in humans and have been recognized as emerging pathogens. These bacteria are mainly transmitted by blood-sucking arthropods, such as fleas and lice. The role of ticks in the transmission of Bartonella spp. is unclear. METHODS: A recently developed quadruplex polymerase chain reaction (PCR) amplicon next-generation sequencing approach that targets Bartonella-specific fragments on gltA, ssrA, rpoB, and groEL was applied to test host-seeking Ixodes scapularis ticks (n = 1641; consisting of 886 nymphs and 755 adults) collected in 23 states of the eastern half of the United States and Ixodes pacificus ticks (n = 966; all nymphs) collected in California in the western United States for the presence of Bartonella DNA. These species were selected because they are common human biters and serve as vectors of pathogens causing the greatest number of vector-borne diseases in the United States. RESULTS: No Bartonella DNA was detected in any of the ticks tested by any target. CONCLUSIONS: Owing to the lack of Bartonella detection in a large number of host-seeking Ixodes spp. ticks tested across a broad geographical region, our results strongly suggest that I. scapularis and I. pacificus are unlikely to contribute more than minimally, if at all, to the transmission of Bartonella spp.

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.

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.

Ixodes scapularis (the blacklegged tick) was considered a species of no medical concern until the mid-1970s. By that time, the tick's geographic distribution was thought to be mainly in the southeastern United States (US), with additional localized populations along the Eastern Seaboard north to southern Massachusetts and in the Upper Midwest. Since 1975, I. scapularis has been implicated as a vector of seven human pathogens and is now widely distributed across the eastern US up to the border with Canada. Geographic expansion of tick-borne diseases associated with I. scapularis (e.g., Lyme disease, anaplasmosis, and babesiosis) is attributed to an expanding range of the tick. However, due to changes in tick surveillance efforts over time, it is difficult to differentiate between range expansion and increased recognition of already established tick populations. We provide a history of the documented occurrence of I. scapularis in the US from its description in 1821 to present, emphasizing studies that provide evidence of expansion of the geographic distribution of the tick. Deforestation and decimation of the white-tailed deer (Odocoileus virginianus), the primary reproductive host for I. scapularis adults, during the 1800s presumably led to the tick disappearing from large areas of the eastern US where it previously had been established. Subsequent reforestation and deer population recovery, together with recent climate warming, contributed to I. scapularis proliferating in and spreading from refugia where it had persisted into the early 1900s. From documented tick collection records, it appears I. scapularis was present in numerous locations in the southern part of the eastern US in the early 1900s, whereas in the north it likely was limited to a small number of refugia sites during that time period. There is clear evidence for established populations of I. scapularis in coastal New York and Massachusetts by 1950, and in northwestern Wisconsin by the late 1960s. While recognizing that surveillance for I. scapularis increased dramatically from the 1980s onward, we describe multiple instances of clearly documented expansion of the tick's geographic distribution in the Northeast, Upper Midwest, and Ohio Valley regions from the 1980s to present. Spread and local population increase of I. scapularis, together with documentation of Borrelia burgdorferi sensu stricto in host-seeking ticks, was universally followed by increases in Lyme disease cases in these areas. Southward expansion of northern populations of I. scapularis, for which the host questing behavior of the nymphal stage leads to substantially higher risk of human bites compared with southern populations, into Virginia and North Carolina also was followed by rising numbers of Lyme disease cases. Ongoing surveillance of ticks and tick-borne pathogens is essential to provide the data needed for studies that seek to evaluate the relative roles of land cover, tick hosts, and climate in explaining and predicting geographic expansion of ticks and tick-borne diseases.

BACKGROUND: Numerous bioassay methods have been used to test the efficacy of repellents for ticks, but the comparability of results across different methods has only been evaluated in a single study. Of particular interest are comparisons between bioassays that use artificial containers (in vitro) with those conducted on a human subject (in vivo) for efficacy testing of new potential unregistered active ingredients, which most commonly use in vitro methods. METHODS: We compared four different bioassay methods and evaluated three ingredients (DEET [N,N-Diethyl-meta-toluamide], peppermint oil and rosemary oil) and a negative control (ethanol) over a 6-h period. Two of the methods tested were in vivo bioassay methods in which the active ingredient was applied to human skin (finger and forearm bioassays), and the other two methods were in vitro methods using artificial containers (jar and petri dish bioassays). All four bioassays were conducted using Ixodes scapularis nymphs. We compared the results using nymphs from two different tick colonies that were derived from I. scapularis collected in the US states of Connecticut and Rhode Island (northern origin) and Oklahoma (southern origin), expecting that ticks of different origin would display differences in host-seeking behavior. RESULTS: The results between bioassay methods did not differ significantly, even when comparing those that provide the stimulus of human skin with those that do not. We also found that tick colony source can impact the outcome of repellency bioassays due to differences in movement speed; behavioral differences were incorporated into the assay screening. DEET effectively repelled nymphs for the full 6-h duration of the study. Peppermint oil showed a similar repellent efficacy to DEET during the first hour, but it decreased sharply afterwards. Rosemary oil did not effectively repel nymphs across any of the time points. CONCLUSIONS: The repellency results did not differ significantly between the four bioassay methods tested. The results also highlight the need to consider the geographic origin of ticks used in repellency bioassays in addition to species and life stage. Finally, our results indicate a limited repellent efficacy of the two essential oils tested, which highlights the need for further studies on the duration of repellency for similar botanically derived active ingredients and for evaluation of formulated products.

The Asian longhorned tick, Haemaphysalis longicornis, an invasive tick species in the United States, has been found actively host-seeking while infected with several human pathogens. Recent work has recovered large numbers of partially engorged, host-seeking H. longicornis, which together with infection findings raises the question of whether such ticks can reattach to a host and transmit pathogens while taking additional bloodmeals. Here we conducted molecular blood meal analysis in tandem with pathogen screening of partially engorged, host-seeking H. longicornis to identify feeding sources and more inclusively characterize acarological risk. Active, statewide surveillance in Pennsylvania from 2020 to 2021 resulted in the recovery of 22/1,425 (1.5%) partially engorged, host-seeking nymphal and 5/163 (3.1%) female H. longicornis. Pathogen testing of engorged nymphs detected 2 specimens positive for Borrelia burgdorferi sensu lato, 2 for Babesia microti, and 1 co-infected with Bo. burgdorferi s.l. and Ba. microti. No female specimens tested positive for pathogens. Conventional PCR blood meal analysis of H. longicornis nymphs detected avian and mammalian hosts in 3 and 18 specimens, respectively. Mammalian blood was detected in all H. longicornis female specimens. Only 2 H. longicornis nymphs produced viable sequencing results and were determined to have fed on black-crowned night heron, Nycticorax nycticorax. These data are the first to molecularly confirm H. longicornis partial blood meals from vertebrate hosts and Ba. microti infection and co-infection with Bo. burgdorferi s.l. in host-seeking specimens in the United States, and the data help characterize important determinants indirectly affecting vectorial capacity. Repeated blood meals within a life stage by pathogen-infected ticks suggest that an understanding of the vector potential of invasive H. longicornis populations may be incomplete without data on their natural host-seeking behaviors and blood-feeding patterns in nature.

Rapid environmental change in Alaska and other regions of the Arctic and sub-Arctic has raised concerns about increasing human exposure to ticks and the pathogens they carry. We tested a sample of ticks collected through a combination of passive and active surveillance from humans, domestic animals, and wildlife hosts in Alaska for a panel of the most common tick-borne pathogens in the contiguous United States to characterize the diversity of microbes present in this region. We tested 189 pooled tick samples collected in 2019-2020 for Borrelia spp., Anaplasma spp., Ehrlichia spp., and Babesia spp. using a multiplex PCR amplicon sequencing assay. We found established populations of Ixodes angustus Neumann (Acari: Ixodidae), Ixodes uriae White (Acari: Ixodidae), and Haemaphysalis leporispalustris Packard (Acari: Ixodidae) in Alaska, with I. angustus found on a variety of hosts including domestic companion animals (dogs and cats), small wild mammals, and humans. Ixodes angustus were active from April through October with peaks in adult and nymphal activity observed in summer months (mainly July). Although no known human pathogens were detected, Babesia microti-like parasites and candidatus Ehrlichia khabarensis were identified in ticks and small mammals. The only human pathogen detected (B. burgdorferi s.s.) was found in a tick associated with a dog that had recently traveled to New York, where Lyme disease is endemic. This study highlights the value of a combined passive and active tick surveillance system to detect introduced tick species and pathogens and to assess which tick species and microbes are locally established.

The Centers for Disease Control and Prevention's national tick and tick-borne pathogen surveillance program collects information to better understand the regional distribution, prevalence, and exposure risk of host-seeking medically important ticks in the United States. A recently developed next generation sequencing (NGS) targeted multiplex PCR amplicon sequencing (MPAS) assay has enhanced the detection capabilities for Ixodes-associated human pathogens found in Ixodes scapularis and Ixodes pacificus ticks compared to the routinely used real-time PCR assay. To operationalize the MPAS assay for the large number of tick surveillance submissions processed each year, a reproducible high throughput bioinformatics pipeline is needed. We describe the development and validation of the MPAS pipeline, a bioinformatics pipeline that identifies and summarizes amplicon sequences produced by the MPAS assay. This pipeline is portable and reproducible across different computing environments, and flexible by allowing modifications to input parameters, assay primer and reference sequences. The automation of the summary report, BLAST report, and phylogenetic analysis reduces the amount of time needed for downstream analysis. To validate this pipeline, we compared the analysis of a MPAS assay dataset consisting of 175 I. scapularis nymphs with the MPAS pipeline and previously published results analyzed with a CLC Genomic Workbench workflow. The MPAS pipeline identified the same number of positive ticks for Anaplasma phagocytophilum and Babesia species as the original analysis, but the MPAS pipeline provided enhanced sequencing resolution of Borrelia burgdorferi sensu lato co-infected samples. The reproducibility, flexibility, analysis automation, and improved sequence resolution of the MPAS pipeline make it well suited for a high throughput tick pathogen surveillance program.

In 2011, Ehrlichia muris eauclairensis (EME) was described as a human pathogen spread by the blacklegged tick, Ixodes scapularis. Until very recently, its reported distribution was limited to the upper midwestern United States, mainly in Minnesota and Wisconsin. In this study, we report the detection of EME DNA in 4 of 16,146 human biting I. scapularis ticks submitted from Massachusetts to a passive tick surveillance program. Active tick surveillance yielded evidence of EME local transmission in the northeastern United States through detection of EME DNA in 2 of 461 host-seeking I. scapularis nymphs, and in 2 white-footed mice (Peromyscus leucopus) of 491 rodent samples collected in the National Ecological Observatory Network (NEON) Harvard Forest site in Massachusetts.

As the primary vector of Lyme disease spirochetes and several other medically significant pathogens, Ixodes scapularis presents a threat to public health in the United States. The incidence of Lyme disease is growing rapidly in upper midwestern states, particularly Michigan, Minnesota, and Wisconsin. The probability of a tick bite, acarological risk, is affected by the phenology of host-seeking I. scapularis. Phenology has been well-studied in northeastern states, but not in the Upper Midwest. We conducted biweekly drag sampling across 4 woodland sites in Minnesota between April and November from 2015 to 2017. The majority of ticks collected were I. scapularis (82%). Adults were active throughout our entire 8-month collection season, with sporadic activity during the summer, larger peaks in activity observed in April, and less consistent and lower peaks observed in October. Nymphs were most active from May through August, with continuing low-level activity in October, and peak activity most commonly observed in June. The observed nymphal peak corresponded with the typical peak in reported human Lyme disease and anaplasmosis cases. These findings are consistent with previous studies from the Upper Midwest and highlight a risk of human exposure to I. scapularis at least from April through November. This information may aid in communicating the seasonality of acarological risk for those living in Minnesota and other upper midwestern states as well as being relevant to the assessment of the ecoepidemiology of Lyme disease and the modeling of transmission dynamics.