Host-utilization differences between larval and nymphal deer ticks in northeastern U.S. sites enzootic for Borrelia burgdorferi sensu stricto.

In the northeastern U.S., Borrelia burgdorferi sensu stricto, the agent of Lyme disease, is maintained between vertebrate hosts and subadult deer ticks (the northern clade of Ixodes scapularis, formerly known as Ixodes dammini). Theoretical arguments suggest that the force of transmission would be greatest when infected nymphal ticks focus their bites on the same host as the uninfected larvae. Stage-specific differences in host utilization would reduce the force of transmission, but to date such differences remain understudied. We determined the host utilization differences of larval and nymphal deer ticks using bloodmeal analysis of host-seeking nymphs and adults collected from 5 field sites in New England. Matched cohorts of ticks (nymphs=506, adults=451), i.e. ticks that had fed during the same summer season, were used to control for yearly host population variations. Infection status of all ticks was determined by real time PCR. Nymphal deer ticks were more likely to have fed on birds and sciurids (13% vs 3%, and 41% vs 9%, respectively p<0.001) and larvae were more likely to have fed on shrews (26% vs 3%, p<0.001). Similarly, ticks that had fed on a mouse or a shrew as larvae were likely to become infected (OR= 3.195, 95% CI [1.9, 5.1] and OR=2.5[1.6,3.8] respectively), and they were positively associated with infection prevalence at our sites. However, very few nymphs fed on shrews, and they were not associated with infection, raising the question of how uninfected shrews acquire infection each year. Sciurids did not appear to contribute to the enzootic cycle at our sites, which may be due to the low numbers of larvae that fed on them. Sciurid-fed ticks of either stage were not associated with infection. Both stages of ticks were less likely to be infected if they had fed on deer (OR=0.08 [0.02.0.3] and OR=0.4 [0.2,0.7] tested as nymphs and adults, respectively) and thus deer likely served to reduce the force of transmission at our sites. Site-specific analysis of differential host utilization by subadult deer ticks may contribute to appropriate targeting of interventions and thereby promote reducing risk of acquiring Lyme disease and the other deer tick-transmitted infections.

Identifying Trusted Sources of Lyme Disease Prevention Information Among Internet Users Connected to Academic Public Health Resources: Internet-Based Survey Study.

BACKGROUND: Misinformation about Lyme disease and other tick-transmitted pathogens circulates frequently on the internet and can compete with, or even overshadow, science-based guidance on tick-borne disease (TBD) prevention. OBJECTIVE: We surveyed internet users connected to academic tick-related resources to identify trusted sources of Lyme disease prevention information, explore confidence in tick bite prevention information, and examine associations of these responses with answers to commonly disputed issues. METHODS: The survey was conducted through social media and website pages for Western Connecticut State University Tickborne Disease Prevention Laboratory and the University of Rhode Island TickEncounter Resource Center. RESULTS: Respondents (N=1190) were predominantly female (903/1190, 76.3%), middle-aged (574/1182, 48.6%), and resided in New England states (663/1190, 55.7%). In total 984 of 1186 (83%) respondents identified conventional experts (eg, the Centers for Disease Control [CDC] or other government health agencies, physicians who follow Infectious Diseases Society of America guidelines for Lyme disease treatment guidelines, and academics) as trustworthy TBD prevention resources. However, nearly one-fourth of respondents would first consult personal contacts and web-based communities regarding prevention information before consulting conventional expert sources. The opinions of public health experts and physicians were rated among the top motivators underlying personal prevention decisions; yet, more than 50% of participants revealed distrustful attitudes toward, or were uncertain about, CDC-supported statements related to time to transmission of Lyme disease (708/1190, 59.5%), the safety of diethyltoluamide-based repellents for children (604/1183, 51.1%), and recommended use of antibiotic prophylaxis (773/1181, 65.4%). Multimodal regression models revealed that participants from high-Lyme-disease-incidence states were more likely to first seek TBD prevention information from personal networks and nontraditional sources before approaching conventional sources of TBD prevention information. We found that those reporting high rates of social media usage were more than twice as likely to first seek traditional expert sources of prevention information but were overall more likely to reject CDC-promoted Lyme disease information, in particular the established time to transmission of Lyme disease bacteria. Models also predicted that those participants who disagreed with the conventional scientific view on the antibiotic prophylaxis prevention statement were less likely to be confident in their ability to protect themselves from a tick bite. Overall, uncertainty in one's ability to protect oneself against tick bites was strongly associated with uncertainty about beliefs in CDC-promoted TBD prevention information. Self-reported trust in experts and frequency of social media use suggest that these platforms may provide opportunities to engage directly with the public about TBD prevention practices. CONCLUSIONS: Using strategies to improve public trust and provide information where the public engages on social media may improve prevention communication and adoption of best practices.

Ixodes scapularis Is the Most Susceptible of the Three Canonical Human-Biting Tick Species of North America to Repellent and Acaricidal Effects of the Natural Sesquiterpene, (+)-Nootkatone.

Ticks are vectors of many human and animal zoonotic disease-causing agents causing significant global health and economic strain. Repellents and acaricides are integral to the human capacity for personal protection from tick bites. Nootkatone, a naturally occurring sesquiterpene found in the Alaskan cedar tree, grapefruit, and other sources, has been documented to be a potent acaricide. Research has also noted repellent effects against some tick species. In this study, our aim was to investigate the effect of synthetic, high-purity (+)-nootkatone on adult Ixodes scapularis, Dermacentor variabilis, and Amblyomma americanum ticks in an in vitro, vertical filter paper bioassay. (+)-nootkatone showed compelling tick repellency, but median effective concentrations (EC50) significantly differed among species. Ixodes scapularis were repelled at very low concentrations (EC50 = 0.87 ± 0.05 µg/cm2). Higher concentrations were required to repel D. variabilis (EC50 = 252 ± 12 µg/cm2) and A. americanum (EC50 = 2313 ± 179 µg/cm2). Significant post-exposure mortality, assessed 24 h after repellency trials, was also observed in I. scapularis but was absent entirely in D. variabilis and A. americanum. These tests demonstrate that nootkatone has a promising dual-action personal protection capacity against adult I. scapularis ticks, warranting further investigation in more natural environments and in the presence of host cues.

Tick-Borne Disease Prevention Behaviors Among Participants in a Tick Surveillance System Compared with a Sample Of Master Gardeners.

Theory-based approaches to health communication and behavior are increasingly applied to interventions that address poor public tick-borne disease prevention knowledge and practices. We sought to understand the tick-borne disease prevention behaviors among participants in a crowdsourced passive tick surveillance system that employs theory-based messages about tick bite risk and prevention strategies. We administered an electronic survey to a randomly selected sample of passive surveillance system users and compared their responses to those from a nationwide sample of Master Gardeners (MG), a group with heighten tick exposure due to outdoor activity. Over 80% of TickSpotters respondents, and over 75% of MG respondents encountered a tick in the past year. Among both groups, tick checks were the most frequently practiced prevention behavior, with over 70% of people performing them most or all the time after outdoor activity. A greater proportion of MGs used skin repellents such as DEET or picaridin than TickSpotters users, but more than 70% of respondents from both groups reported that they never or only sometimes use permethrin-treatment on clothing, and nearly half of both groups reportedly used no peridomestic tick treatments. TickSpotters respondents overwhelmingly reported recording tick encounter information and saving specimens for identification and testing, while only a small percentage of MGs monitored their tick encounters. These findings suggest that while both TickSpotters and MG groups appear to be practicing some important tick bite prevention behaviors, there remain areas that could benefit from targeted theory-based interventional approaches.

Identification of Hard Ticks in the United States: A Practical Guide for Clinicians and Pathologists.

ABSTRACT: According to guidelines published by the Infectious Disease Society of America, Lyme disease prophylaxis is possible if a tick can be identified as Ixodes scapularis (nymphal or adult) within 72 hours of tick removal. However, a recent survey of medical practitioners indicates generally poor proficiency in tick identification. In this study, we provide a simple, practical guide to aid medical practitioners in identifying the most commonly encountered human biting ticks of North America.

Proficiency at Tick Identification by Pathologists and Clinicians Is Poor.

OBJECTIVES: Prompt accurate identification of tick species is required for appropriate administration of single dose antimicrobial prophylaxis for Lyme disease in selected patients. To determine the proficiency of clinicians at tick identification in the northeastern United States where Lyme disease has its highest incidence, we undertook a survey. METHODS: We analyzed the results of a voluntary survey testing proficiency in identifying tick species using high-resolution photographs of ticks. RESULTS: Only 35% of ticks were correctly identified. Although 60% of respondents could identify a nonengorged adult blacklegged tick, only 34% could correctly identify a partially engorged blacklegged tick. Participants performed even worse at classifying brown dog, American dog, and Lone Star ticks. CONCLUSIONS: Proficiency of tick identification by pathologists and clinicians is poor.

Assessing Durability and Safety of Permethrin Impregnated Uniforms Used by Outdoor Workers to Prevent Tick Bites after One Year of Use.

Long lasting permethrin-impregnated (LLPI) clothing can retain permethrin and repel ticks for up to three months and without exceeding EPA-approved safe levels; however, little is known about longer term effects of wearing LLPI clothing. Here, permethrin content was measured in new forester pants soon after initial impregnation (Insect Shield) and again one year later after being repeatedly worn by foresters in the field. Urine samples were collected from foresters for biomonitoring of permethrin metabolites at multiple time intervals (pre-use, one-month, three-to-four-months, and one-year post-use). Lethality against nymphal Ixodes scapularis Say was measured in clothing after one year of wear by foresters. Furthermore, to test potential variability in permethrin impregnation of different batches of clothing, separate sets of clothing were anonymously sent to Insect Shield for permethrin treatment over a period of three months and permethrin was quantified. Results demonstrated 33% of participants' pants had no measurable permethrin after one year of wear and permethrin content and tick mortality varied significantly between clothing. Only two of the participants' clothing resulted in ≥ 30% tick mortality after one year of wear. Significant differences were observed in 3-PBA and trans-DCCA, but not cis-DCCA metabolites in participants over the four measured time points and were higher than general United States population levels. This study provides practical information on the safety (measured by urinary metabolites) over time of LLPI clothing. It also provides snapshots (pre-washing and after one year of wear) of effectiveness of LLPI clothing as personal protective equipment against ticks for outdoor workers.

Incrimination of shrews as a reservoir for Powassan virus.

Powassan virus lineage 2 (deer tick virus) is an emergent threat to American public health, causing severe neurologic disease. Its life cycle in nature remains poorly understood. We use a host-specific retrotransposon-targeted real time PCR assay to test the hypothesis that white-footed mice, considered the main eastern U.S. reservoir of the coinfecting agent of Lyme disease, is the reservoir for deer tick virus. Of 20 virus-infected host-seeking nymphal black-legged ticks 65% fed on shrews and none on mice. The proportion of ticks feeding on shrews at a site is positively associated with prevalence of viral infection, but not the Lyme disease agent. Viral RNA is detected in the brain of one shrew. We conclude that shrews are a likely reservoir host for deer tick virus and that host bloodmeal analysis can provide direct evidence to incriminate reservoir hosts, thereby promoting our understanding of the ecology of tick-borne infections.

Potential Effects of Climate Change on Tick-borne Diseases in Rhode Island.

Human cases of tick-borne diseases have been increasing in the United States. In particular, the incidence of Lyme disease, the major vector-borne disease in Rhode Island, has risen, along with cases of babesiosis and anaplasmosis, all vectored by the blacklegged tick. These increases might relate, in part, to climate change, although other environmental changes in the northeastern U.S. (land use as it relates to habitat; vertebrate host populations for tick reproduction and enzootic cycling) also contribute. Lone star ticks, formerly southern in distribution, have been spreading northward, including expanded distributions in Rhode Island. Illnesses associated with this species include ehrlichiosis and alpha-gal syndrome, which are expected to increase. Ranges of other tick species have also been expanding in southern New England, including the Gulf Coast tick and the introduced Asian longhorned tick. These ticks can carry human pathogens, but the implications for human disease in Rhode Island are unclear.

An analysis of companion animal tick encounters as revealed by photograph-based crowdsourced data.

BACKGROUND: Community science is increasingly utilized to track important vectors of companion animal disease, providing a scalable, cost-effective strategy for identifying new foci, changing phenology, and disease prevalence across wide geographies. OBJECTIVES: We examined photographs of ticks found attached to predominately dogs and cats reported to a photograph-based tick surveillance program to identify potential areas for improvements in tick prevention education and risk intervention. METHODS: We compared estimated days of tick attachment using a Kruskal-Wallis one-way analysis of variance, and a Pearson's chi-square analysis of variance on the number of submissions by host type submitted for each season. RESULTS: The blacklegged tick (Ixodes scapularis) was the most common species reported (39.8%). Tick photographs submitted were almost entirely adults (89.5%), and ticks found on companion animals exhibited an estimated median engorgement time of 2.5 days. Ixodes scapularis displayed the highest median engorgement of the top tick species found feeding on companion animals (χ2  = 98.96, p < 0.001). Ticks were spotted year-round; during spring and summer, ticks collected from pets represented 15.4 and 12.8% of all submissions, but increased to 28.5 and 35.2% during autumn and winter, respectively. CONCLUSIONS: Crowdsourced data reveal that mostly adult ticks are detected on pets, and they are found at a point in the blood-feeding process that puts pets at heightened risk for disease transmission. The increase in proportion of ticks found on pets during colder months may reveal a critical knowledge gap amongst pet owners regarding seasonal activity of I. scapularis, a vector of Lyme disease, providing an opportunity for prevention-education.

Crowdsourced Tick Image-Informed Updates to U.S. County Records of Three Medically Important Tick Species.

Burgeoning cases of tick-borne disease present a significant public health problem in the United States. Passive tick surveillance gained traction as an effective way to collect epidemiologic data, and in particular, photograph-based tick surveillance can complement in-hand tick specimen identification to amass distribution data and related encounter demographics. We compared the Federal Information Processing Standards (FIPS) code of tick photos submitted to a free public identification service (TickSpotters) from 2014 to 2019 to published nationwide county reports for three tick species of medical concern: Ixodes scapularis Say (Ixodida: Ixodidae), Ixodes pacificus Cooley and Kohls (Ixodida: Ixodidae), and Amblyomma americanum Linneaus (Ixodida: Ixodidae). We tallied the number of TickSpotters submissions for each tick species according to "Reported" or "Established" criteria per county, and found that TickSpotters submissions represented more than half of the reported counties of documented occurrence, and potentially identified hundreds of new counties with the occurrence of these species. We detected the largest number of new county reports of I. scapularis presence in Michigan, North Carolina, and Texas. Tick image submissions revealed potentially nine new counties of occurrence for I. pacificus, and we documented the largest increase in new county reports of A. americanum in Kentucky, Illinois, Indiana, and Ohio. These findings demonstrate the utility of crowdsourced photograph-based tick surveillance as a complement to other tick surveillance strategies in documenting tick distributions on a nationwide scale, its potential for identifying new foci, and its ability to highlight at-risk localities that might benefit from tick-bite prevention education.

Evaluating the Effects of Minimal Risk Natural Products for Control of the Tick, Ixodes scapularis (Acari: Ixodidae).

Knockdown and residual activity of 10 minimal risk natural products (MRNPs), one experimental formulation of nootkatone, and two bifenthrin labels were evaluated against host-seeking nymphal Ixodes scapularis Say using a novel micro-plot product screening system placed in a landscape setting similar to a wooded residential property. The MRNPs evaluated included Tick Stop, EcoPCO EC-X, Met52 EC, CedarCide PCO Choice, EcoEXEMPT IC2, EcoSMART Organic Insecticide, Essentria IC3, privately labeled products 1 and 2 (based on EcoEXEMPT IC2 and sold as a professional pest control application), and Tick Killz. Just the nootkatone and 4 of these 10 products tested (EcoPCO EC-X, Met52 EC, EcoEXEMPT IC2, and Essentria IC3) had statistically significant (P < 0.05) knockdown effects (killed ticks while active in the arenas) when compared to water-only controls, but only 2 of these, EcoPCO EC-X and nootkatone, displayed significant residual tick-killing activity after weathering naturally in the landscape for 2 wk prior to tick application/testing. Moreover, botanical oil-based products with the same active ingredients provided inconsistent results when tested multiple times across study years.

Assessing Public Tick Identification Ability and Tick Bite Riskiness Using Passive Photograph-Based Crowdsourced Tick Surveillance.

Tick identification is critical for assessing disease risk from a tick bite and for determining requisite treatment. Data from the University of Rhode Island's TickEncounter Resource Center's photo-based surveillance system, TickSpotters, indicate that users incorrectly identified their submitted specimen 83% of the time. Of the top four most commonly submitted tick species, western blacklegged ticks (Ixodes pacificus Cooley & Kohls [Ixodida: Ixodidae]) had the largest proportion of unidentified or misidentified submissions (87.7% incorrectly identified to species), followed by lone star ticks (Amblyomma americanum Linneaus [Ixodida: Ixodidae]; 86.8% incorrect), American dog ticks (Dermacentor variabilis Say [Ixodida: Ixodidae]; 80.7% incorrect), and blacklegged ticks (Ixodes scapularis Say [Ixodida: Ixodidae]; 77.1% incorrect). More than one quarter of participants (26.3%) submitted photographs of ticks that had been feeding for at least 2.5 d, suggesting heightened risk. Logistic regression generalized linear models suggested that participants were significantly more likely to misidentify nymph-stage ticks than adult ticks (odds ratio [OR] = 0.40, 95% confidence interval [CI]: 0.23, 0.68, P < 0.001). Ticks reported on pets were more likely to be identified correctly than those found on humans (OR = 1.07, 95% CI: 1.01-2.04, P < 0.001), and ticks feeding for 2.5 d or longer were more likely to be misidentified than those having fed for one day or less (OR = 0.43, 95% CI: 0.29-0.65, P < 0.001). State and region of residence and season of submission did not contribute significantly to the optimal model. These findings provide targets for future educational efforts and underscore the value of photograph-based tick surveillance to elucidate these knowledge gaps.

Retrotransposon-Based Blood Meal Analysis of Nymphal Deer Ticks Demonstrates Spatiotemporal Diversity of Borrelia burgdorferi and Babesia microti Reservoirs.

Deer tick-transmitted Borrelia burgdorferisensu stricto (Lyme disease) and Babesia microti (babesiosis) increasingly burden public health across eastern North America. The white-footed mouse is considered the primary host for subadult deer ticks and the most important reservoir host for these and other disease agents. Local transmission is thought to be modulated by less reservoir-competent hosts, such as deer, diverting ticks from feeding on mice. We measured the proportion of mouse-fed or deer-fed host-seeking nymphs from 4 sites during 2 transmission seasons by blood meal remnant analysis using a new retrotransposon-based quantitative PCR (qPCR) assay. We then determined the host that was associated with the infection status of the tick. During the first year, the proportion of mouse-fed ticks ranged from 17% on mainland sites to 100% on an island, while deer-fed ticks ranged from 4% to 24%. The proportion of ticks feeding on mice and deer was greater from island sites than mainland sites (on average, 92% versus 43%). Mouse-fed ticks decreased significantly during year 2 in 3 of 4 sites (most were <20%), while deer-fed ticks increased for all sites (75% at one site). Overall, ticks were more likely to be infected when they had fed on mice (odds ratio [OR] of 2.4 and 1.6 for Borrelia and Babesia, respectively) and were less likely to be infected if they had fed on deer (OR, 0.8 and 0.4). We conclude that host utilization by deer ticks is characterized by significant spatiotemporal diversity, which may confound efficacy tests of interventions targeting reservoir hosts.IMPORTANCE White-footed mice are thought to be the most important reservoir host for the deer tick-transmitted pathogens that cause Lyme disease and human babesiosis because they are the primary host for immature ticks. Transmission would be reduced, however, if ticks feed on deer, which are not capable of infecting ticks with either pathogen. By directly measuring whether ticks had fed on either mice or deer using a new quantitative PCR (qPCR) assay to detect remnants of host DNA leftover from the larval blood meal, we demonstrate that host utilization by ticks varies significantly over time and space and that mice often feed fewer ticks than expected. This finding has implications for our understanding of the ecology of these diseases and for the efficacy of control measures.

Crowdsourced Photographs as an Effective Method for Large-Scale Passive Tick Surveillance.

As tick vector ranges expand and the number of tickborne disease cases rise, physicians, veterinarians, and the public are faced with diagnostic, treatment, and prevention challenges. Traditional methods of active surveillance (e.g., flagging) can be time-consuming, spatially limited, and costly, while passive surveillance can broadly monitor tick distributions and infection rates. However, laboratory testing can require service fees in addition to mailing and processing time, which can put a tick-bite victim outside the window of potential prophylactic options or under unnecessary antibiotic administration. We performed a retrospective analysis of a national photograph-based crowdsourced tick surveillance system to determine the accuracy of identifying ticks by photograph when compared to those same ticks identified by microscopy and molecular methods at a tick testing laboratory. Ticks identified by photograph were correct to species with an overall accuracy of 96.7% (CI: 0.9522, 0.9781; P < 0.001), while identification accuracy for Ixodes scapularis Say (Ixodida: Ixodidae), Amblyomma americanum Linnaeus (Ixodida: Ixodidae), and Dermacentor variabilis Say (Ixodida: Ixodidae), three ticks of medical importance, was 98.2% (Cohen's kappa [κ] = 0.9575; 95% CI: 0.9698, 0.9897), 98.8% (κ = 0.9466, 95% CI: 0.9776, 0.9941), and 98.8% (κ = 0.9515, 95% CI: 0.9776, 0.9941), respectively. Fitted generalized linear models revealed that tick species and stage were the most significant predictive factors that contributed to correct photograph-based tick identifications. Neither engorgement, season, nor location of submission affected identification ability. These results provide strong support for the utility of photograph-based tick surveillance as a tool for risk assessment and monitoring among commonly encountered ticks of medical concern.

First glimpse into the origin and spread of the Asian longhorned tick, Haemaphysalis longicornis, in the United States.

Established populations of Asian longhorned ticks (ALT), Haemaphysalis longicornis, were first identified in the United States (US) in 2017 by sequencing the mitochondrial cytochrome c oxidase subunit I (cox1) 'barcoding' locus followed by morphological confirmation. Subsequent investigations detected ALT infestations in 12, mostly eastern, US states. To gain information on the origin and spread of US ALT, we (1) sequenced cox1 from ALT populations across 9 US states and (2) obtained cox1 sequences from potential source populations [China, Japan and Republic of Korea (ROK) as well as Australia, New Zealand and the Kingdom of Tonga (KOT)] both by sequencing and by downloading publicly available sequences in NCBI GenBank. Additionally, we conducted epidemiological investigations of properties near its initial detection locale in Hunterdon County, NJ, as well as a broader risk analysis for importation of ectoparasites into the area. In eastern Asian populations (China/Japan/ROK), we detected 35 cox1 haplotypes that neatly clustered into two clades with known bisexual versus parthenogenetic phenotypes. In Australia/New Zealand/KOT, we detected 10 cox1 haplotypes all falling within the parthenogenetic cluster. In the United States, we detected three differentially distributed cox1 haplotypes from the parthenogenetic cluster, supporting phenotypic evidence that US ALT are parthenogenetic. While none of the source populations examined had all three US cox1 haplotypes, a phylogeographic network analysis supports a northeast Asian source for the US populations. Within the United States, epidemiological investigations indicate ALT can be moved long distances by human transport of animals, such as horses and dogs, with smaller scale movements on wildlife. These results have relevant implications for efforts aimed at minimizing the spread of ALT in the United States and preventing additional exotic tick introductions.

Molecular Characterization of Haemaphysalis Species and a Molecular Genetic Key for the Identification of Haemaphysalis of North America.

Haemaphysalis longicornis (Acari: Ixodidae), the Asian longhorned tick, is native to East Asia, but has become established in Australia and New Zealand, and more recently in the United States. In North America, there are other native Haemaphysalis species that share similar morphological characteristics and can be difficult to identify if the specimen is damaged. The goal of this study was to develop a cost-effective and rapid molecular diagnostic assay to differentiate between exotic and native Haemaphysalis species to aid in ongoing surveillance of H. longicornis within the United States and help prevent misidentification. We demonstrated that restriction fragment length polymorphisms (RFLPs) targeting the 16S ribosomal RNA and the cytochrome c oxidase subunit I (COI) can be used to differentiate H. longicornis from the other Haemaphysalis species found in North America. Furthermore, we show that this RFLP assay can be applied to Haemaphysalis species endemic to other regions of the world for the rapid identification of damaged specimens. The work presented in this study can serve as the foundation for region specific PCR-RFLP keys for Haemaphysalis and other tick species and can be further applied to other morphometrically challenging taxa.

Is selenoprotein K required for Borrelia burgdorferi infection within the tick vector Ixodes scapularis?

BACKGROUND: Tick selenoproteins are involved in regulating oxidative and endoplasmic reticulum stress during prolonged tick feeding on mammalian hosts. How selenoproteins are activated upon tick-borne pathogen infection is yet to be defined. METHODS: To examine the functional role of selenoprotein K in Borrelia burgdorferi infection within the tick host Ixodes scapularis, RNA interference (RNAi)-based gene silencing was performed. RESULTS: Selenoprotein K is an endoplasmic reticulum (ER)-resident protein and a component of the ERAD complex involved in ER homeostasis. A qRT-PCR assay revealed the significant upregulation of selenogene K (selenoK) expression in B. burgdorferi-infected tick tissues. Silencing of the selenoK transcript significantly depleted B. burgdorferi copies within the infected tick tissues. Upon selenoK knockdown, another component of the ERAD complex, selenoprotein S (selenoS), was significantly upregulated, suggesting a compensatory mechanism to maintain ER homeostasis within the tick tissues. Knockdown of selenoK also upregulated ER stress-related unfolded protein response (UPR) pathway components, ATF6 and EIF2. CONCLUSIONS: The exact mechanisms that contribute to depletion of B. burgdorferi upon selenoK knockdown is yet to be determined, but this study suggests that selenoK may play a vital role in the survival of B. burgdorferi within the tick host.

Bioabsorption and effectiveness of long-lasting permethrin-treated uniforms over three months among North Carolina outdoor workers.

BACKGROUND: Vector-borne diseases are an important cause of morbidity and mortality in the USA. Effective, convenient prevention methods are needed. Long-lasting permethrin-impregnated (LLPI) clothing can prevent tick bites, however, additional information is needed on the real-world effectiveness and safety of this preventative measure. METHODS: In this pilot study, we recruited state and county park employees from North Carolina to wear LLPI uniforms for three months during the summer of 2016. We collected spot urine samples for biomonitoring of permethrin metabolites at one week, one month and three months after first use of the LLPI uniform. Following three months of wear, we collected pants and socks and analyzed them for permethrin content and mortality to ticks and mosquitoes. RESULTS: Thirteen park employees were included in the analysis. Bioactive amounts of permethrin remained in all clothing swatches tested, although there was great variability. Tick mortality was high, with 78% of pant and 88% of sock swatches having mean knockdown percentages ≥ 85%. In contrast, mosquito mortality was low. Over the study period, the absorbed dosage of permethrin averaged < 4 µg/kg/d of body weight based on measurements of three metabolites. CONCLUSIONS: LLPI clothing retained permethrin and bioactivity against ticks after three months of use in real-world conditions. The estimated absorbed dosage of permethrin was well below the U.S. EPA level of concern, suggesting that LLPI clothing can be used safely by outdoor workers for tick bite prevention.