Tick-borne diseases in Europe: Current prevention, control tools and the promise of aptamers.

In Europe, tick-borne diseases (TBDs) cause significant morbidity and mortality, affecting both human and animal health. Ticks can transmit a wide variety of pathogens (bacteria, viruses, and parasites) and feed on many vertebrate hosts. The incidence and public health burden of TBDs are tending to intensify in Europe due to various factors, mainly anthropogenic and often combined. Early detection of tick-borne pathogens (TBPs), preventive measures and treatment are of great importance to control TBDs and their expansion. However, there are various limitations in terms of the sensitivity and/or specificity of detection and prevention methods, and even in terms of feasibility. Aptamers are single-stranded DNA or RNA that could address these issues as they are able to bind with high affinity and specificity to a wide range of targets (e.g., proteins, small compounds, and cells) due to their unique three-dimensional structure. To date, aptamers have been selected against TBPs such as tick-borne encephalitis virus, Francisella tularensis, and Rickettsia typhi. These studies have demonstrated the benefits of aptamer-based assays for pathogen detection and medical diagnosis. In this review, we address the applications of aptamers to TBDs and discuss their potential for improving prevention measures (use of chemical acaricides, vaccination), diagnosis and therapeutic strategies to control TBDs.

Subolesin knockdown in tick cells provides insights into vaccine protective mechanisms.

Ticks as obligate blood-feeding arthropod vectors of pathogenic viruses, bacteria, protozoa and helminths associated with prevalent tick-borne diseases (TBDs) worldwide. These arthropods constitute the second vector after mosquitoes that transmit pathogens to humans and the first vector in domestic animals. Vaccines constitute the safest and more effective approach to control tick infestations and TBDs, but research is needed to identify new antigens and improve vaccine formulations. The tick protein Subolesin (Sub) is a well-known vaccine protective antigen with a highly conserved sequence at both gene and protein levels in the Ixodidae and among arthropods and vertebrates. In this study, transcriptomics and proteomics analyses were conducted together with graph theory data analysis in wild type and Sub knockdown (KD) tick ISE6 cells in order to identify and characterize the functional implications of Sub in tick cells. The results support a key role for Sub in the regulation of gene expression in ticks and the relevance of this antigen in vaccine development against ticks and TBDs. Proteins with differential representation in response to Sub KD provide insights into vaccine protective mechanisms and candidate tick protective antigens.

A study of knowledge, attitudes, and practices on ticks and tick-borne diseases of cattle among breeders of two bovine Portuguese autochthonous breeds.

Beef cattle production in Portugal is an important sector of national agricultural production, with half of the herd being in the Alentejo region. Despite this, animal health is essential for its productivity, which may be compromised by ticks and tick-borne diseases. So far, no study has been conducted in Portugal to assess knowledge, attitudes, and practices (KAP) on ticks and tick-borne diseases in cattle, which the authors are aware of. This type of questionnaire is a very useful tool in the development and application of effective and sustainable prevention and control measures. Therefore, a KAP questionnaire was applied to 44 cattle breeders of autochthonous Portuguese breeds, namely 14 breeders of the Alentejana breed and 30 of the Mertolenga breed, between January 1 and May 9, 2023. Based on the analysis criteria of these surveys, 64% of the Alentejana breeders and 63% of the Mertolenga breeders have an average level of knowledge about ticks and tick-borne diseases, and 21% of the Alentejana breeders and 33% of the Mertolenga breeders have a high level of knowledge. Although only 21.4% of the Alentejana and 36.7% of the Mertolenga breeders consider tick infestation as a major animal health problem, 71.4% of the Alentejana and 63.3% of breeders of the Mertolenga state that one of the main reasons for veterinary consultations on their farm is deworming of animals, and 92.9% of breeders of the Alentejana and 96.7% of breeders of the Mertolenga refer the use of dewormers as a strategy to control tick infestation. The results of this study contribute to highlighting the importance of correcting some identified knowledge gaps and improving knowledge, especially on the life cycle of this parasite, its local distribution and seasonality, resistance to acaricides, and alternative control strategies.

[Anthropization and tick-borne diseases: the example of Lyme borreliosis]. / Anthropisation et maladies à tique : l'exemple de la borréliose de Lyme.

Ticks and tick-borne diseases are on the rise throughout the world. The reasons are multifactorial but all associated with human practices, including climate change and socio-economic and eco systemic changes. In the northern hemisphere, Lyme borreliosis and its vector, the tick belonging to the Ixodes ricinus complex, are particularly studied. Changes in forestry and the expansion of certain wild ungulates since the Second World War could explain the increasing presence of this tick in our environment. As it is likely to transmit other microorganisms potentially pathogenic to humans, an integrated multidisciplinary approach to identify human practices promoting its expansion is critical to control the (re)emergence of infectious diseases. Other ticks also benefit from the same anthropised context to increase their numbers in the environment.

Les tiques et maladies à tiques sont en expansion à travers le monde. Les raisons sont multifactorielles mais toutes associées aux pratiques humaines, notamment les modifications climatiques et les changements socio-économiques et écosystémiques. Dans l'hémisphère nord, la borréliose de Lyme et son vecteur, la tique appartenant au complexe Ixodes ricinus sont particulièrement étudiés. Les modifications de sylviculture et l'expansion de certains ongulés sauvages depuis la seconde guerre mondiale pourraient expliquer la présence de plus en plus importante de la tique dans notre environnement. Cette tique étant susceptible de transmettre d'autres microorganismes potentiellement pathogènes pour l'homme, une approche multidisciplinaire intégrée afin d'identifier les pratiques humaines qui favorisent son expansion est essentielle afin de contrôler la (ré)-émergence de maladies infectieuses. D'autres tiques bénéficient du même contexte anthropisé pour augmenter leur nombre dans l'environnement.

An integrative framework for tick management: the need to connect wildlife science, One Health, and interdisciplinary perspectives.

Vector-borne diseases pose a significant threat to human and animal health worldwide. Their emergence is influenced by various factors such as environmental changes, host characteristics, and human behavior. The One Health approach is necessary to thoroughly investigate tick-borne diseases and understand the complex interactions between environmental, animal, and human health. Anthropogenic changes have impacted predators, leading to cascading effects on wildlife prey species and the emergence of vector-borne diseases. The increase in global trade and travel has led to the introduction of several invasive vector species, increasing the risk of zoonotic pathogen spillover. Tick and tick-borne disease research requires an interdisciplinary approach to address challenges in a One Health paradigm.

Knowledge, Attitudes, and Practices Regarding Ticks, Tick-Borne Diseases, and Ethnomedicine Among an at-Risk Population in Kerala.

Tick and tick-borne diseases (TBDs) are increasing annually, and the study of ticks has gained importance after the outbreak of Kyasanur Forest disease (KFD) in the South Western Ghats of India. Wayanad district of Kerala, with the highest tribal population in South India, is a KFD endemic state, owing to the lack of knowledge, attitude, and practice studies on TBDs and ethnomedicines against ticks. This study was carried out to assess their baseline knowledge, attitude, and ethnomedicinal practice against ticks. A structured questionnaire was used to conduct a survey of 499 tribal members living in forest fringe areas. Logistic regression analysis was performed to identify the factors that influence the knowledge, attitude, and practice of tribes on TBDs. More than 70% of the population visit the forests on a regular basis, with 65.7% of the population regularly exposed to tick bites; however, only 47.7% were aware of TBDs. About 47.4% of the respondents took precautions like therapeutics and natural remedies to avoid tick bites. Ten species of medicinal plant belonging to eight different families have been identified from the survey. The tribal population use these plants to repel ticks as well as treat tick bites. From the study, we concluded that the limited in-depth knowledge displayed by the tribes can be strengthened by conducting community programs such as awareness classes on TBD and its control measures. The ethnobotanicals identified can be used to formulate novel tick repellents in the future.

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

CONTEXT: 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. PROGRAM: 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. IMPLEMENTATION: 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. EVALUATION: 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. DISCUSSION: 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.

Modelling the burden of disease for cattle-A case of ticks and tick-borne diseases in cattle in a rural set-up in South Africa.

BACKGROUND: The study aimed to estimate the burden of ticks and tick-borne diseases (TBDs) among rural cattle-keeping households of the Eastern Cape province of South Africa using Productivity Adjusted Life Years (PALYs). We modified Disability Adjusted Life Year (DALY) equations for humans to PALYs to estimate the societal burden of tick-borne animal diseases. Whilst the World Health Organization has indicated the adaptability of DALYs to assess burden of animal diseases, nothing has been done in this regard. This could be due to several reasons including that the assessment of animal disease burden is often less of a priority compared to human diseases, particularly in low- and middle-income countries where resources may be limited. As a result, less funding and attention may be given to developing and implementing PALYs for animal diseases. Furthermore, technical and conceptual challenges may be associated with applying DALYs equations to animal diseases, such as determining appropriate measures of productivity loss for different types and categories of animals and diseases. This motivated our study, which is focused on modelling the burden of ticks and TBDs in cattle (cows, oxen and bulls) reared in resource-poor settings. METHOD: We formulated a PALYs approach for cattle populations by adapting the DALYs approach to assess the burden of ticks and TBDs for cattle populations in 20 villages in the Eastern Cape province of South Africa. PALYs is a measurement used to assess the burden of disease in cattle populations, quantifying the years of life lost due to premature mortality and disability. It encompasses years of life lost due to premature mortality (YLL) and years lost due to disability (YLD) caused by health conditions. PALYs provide a comprehensive perspective on the effective number of years lost due to disability and premature death in cattle populations. The PALYs model involves several parameters that are examined to understand their impact on the model's behavior. To illustrate this, we used a structured questionnaire to collect data on parameters that feed into PALY equations. We coded and entered data from questionnaires directly into Statistical Package of Social Sciences (IBM SPSS Version 20) and entered the estimated values of PALY parameters to calculate PALYs equations, which were to estimate the societal disease burden of ticks and TBDs in cattle. PALYs calculations were done in three categories; PALYs without discounting and age weighting, PALYs with only discounting, and PALYs with discounting and age weighting in a practical example to study how these parameters influence the outcomes of the PALYs model. RESULTS: Our results revealed that the years of productivity lost by a cow, bull, and ox that suffered from ticks and TBDs could be estimated at various disability weights. Approximately 26%, 23% and 35% of the productivity years of a cow, ox and bull, respectively, reared by resource-poor livestock owners are lost due to the burden of ticks and TBDs in the Eastern Cape province of South Africa. However, introducing tick control measures reduces the loss to approximately 3%, 2% and 3% of their lifespan productivity, an indication that tick control will save approximately 23%, 21% and 32% of years of the productive life of cows, oxen and bulls, respectively. Therefore, it is evident that using ticks and TBD prevention measures at an early age of cattle will improve cattle productivity and hence the socioeconomic welfare of resource-poor rural farming communities in the Eastern Cape province of South Africa. CONCLUSION: The findings generated from the PALYs approach are helpful in projections for the future burden of any livestock disease. They may be used as a basis in policy formulation and decision-making by various stakeholders, and hence a priority in animal health economics. We recommend that a classification of livestock diseases of national economic importance should consider both the societal burden (non-monetary) and economic impact instead of the common practice of only considering the economic (monetary) impact. Adding a societal burden measure to existing economic measures provides a holistic understanding of the impact of a disease on society especially in resource-limited settings where the livestock value goes beyond monetary value.

Vaccines against tick-borne diseases: a big step forward?

Ticks and tick-borne diseases are on the rise due to socioecosystemic changes and climate modification and are affecting human and animal health. Few vaccines are available. Two recent articles from Matias et al. and Pine et al. used mRNA technology to explore tick and pathogen proteins as vaccine candidates.

Control of ticks and tick-borne diseases in Africa through improved diagnosis and utilisation of data on acaricide resistance.

A meeting, sponsored by the Bill and Melinda Gates Foundation (BMGF) and organised by Clinglobal, was held at The International Livestock Research Institute (ILRI) in Nairobi, Kenya, from 19th - to 21st October 2022. The meeting assembled a unique group of experts on tick control in Africa. Academia, international agencies (FAO and ILRI), the private Animal Health sector and government veterinary services were represented. The significant outcomes included: (i) a shared commitment to standardisation and improvement of acaricide resistance bioassay protocols, particularly the widely used larval packet test (LPT); (ii) development of novel molecular assays for detecting acaricide resistance; (3) creation of platforms for disseminating acaricide resistance data to farmers, veterinary service providers and veterinary authorities to enable more rational evidence-based control of livestock ticks. Implementation of enhanced control will be facilitated by several recently established networks focused on control of parasites in Africa and globally, whose activities were presented at the meeting. These include a newly launched community of practice on management of livestock ticks, coordinated by FAO, an African module of the World Association for the Advancement of Veterinary Parasitology (WAAVP-AN) and the MAHABA (Managing Animal Health and Acaricides for a Better Africa) initiative of Elanco Animal Health.

Innovative approaches for the control of ticks and tick-borne diseases.

Ticks and tick-borne diseases constitute a major threat for human and animal health worldwide. Vaccines for the control of tick infestations and transmitted pathogens still represents a challenge for science and health. Vaccines have evolved with antigens derived from inactivated pathogens to recombinant proteins and vaccinomics approaches. Recently, vaccines for the control of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) have shown the efficacy of new antigen delivery platforms. However, until now only two vaccines based on recombinant Bm86/Bm95 antigens have been registered and commercialized for the control of cattle-tick infestations. Nevertheless, recently new technologies and approaches are under consideration for vaccine development for the control of ticks and tick-borne pathogens. Genetic manipulation of tick commensal bacteria converted enemies into friends. Frankenbacteriosis was used to control tick pathogen infection. Based on these results, the way forward is to develop new paratransgenic interventions and vaccine delivery platforms for the control of tick-borne diseases.

Can protection motivation theory predict protective behavior against ticks?

BACKGROUND: Cases of reported tick-borne diseases in humans have increased over the past decades. Strategies informing the public about ticks, their associated diseases, and preventive measures are often highlighted as important in limiting pathogen transfer and disease. However, knowledge about the motivation for people to apply preventative measures is sparse. METHODS: The aim was to examine if Protection Motivation Theory, a model of disease prevention and health promotion, can predict the use of protective measures against ticks. Ordinal logistic regression and Chi-square tests were used on data from a cross-sectional survey with respondents from Denmark, Norway, and Sweden (n = 2658). We examined the effect of (1) the perceived seriousness of tick bites, Lyme borreliosis (LB), and tick-borne encephalitis (TBE), and (2) the perceived probability of getting a tick bite, Lyme borreliosis, and tick-borne encephalitis on protection against ticks. Finally, we examined if there was an association between the use of a protective measure and the perceived efficacy of that measure. RESULTS: The perceived seriousness of a tick bite and LB significantly predict who is more likely to apply protective measures for all three countries combined. The perceived seriousness of TBE did not significantly predict the level of adoption of protective measures applied by respondents. The perceived likelihood of getting a tick bite within the next 12 months and the perceived likelihood of getting LB if bitten by a tick significantly predicted the application of protective measures. However, the increases in the likelihood of protection were very small. The application of a certain type of protection was always correlated with the perceived efficacy of the same protective measure. CONCLUSION: Some variables of PMT may be used to predict the level of adoption of protection applied against ticks and tick-borne diseases. We found that the perceived seriousness of a tick bite and LB significantly predict the level of adoption protection. The perceived likelihood of getting a tick bite or LB also significantly predicted the level of adoption of protection, although the change was very small. The results regarding TBE were less clear. Lastly, there was an association between applying a protective measure and the perceived efficacy of the same measure.

Citizen Science Informs How Missourians Interact With Ticks.

Understanding which tick species Missourians interact with and where exposures occur can help prevent and diagnose tickborne diseases. A statewide survey of ticks and tickborne pathogens was conducted using citizen science. Missourians submitted 12,819 ticks from April-December 2021. In total, 71% of ticks were Amblyomma americanum, and 89% were collected from a person or a pet. These preliminary data provide tick distributions at the county level and identify patterns of direct tick exposure to humans.

Farmers' Knowledge and Practices About Ticks and Tickborne Diseases in Illinois.

OBJECTIVE: Tickborne diseases (TBDs) in Illinois have increased in recent years. A growing body of literature indicates that the risk of exposure to ticks and tickborne diseases is higher among outdoor workers, including farmers. However, information is lacking on awareness of ticks and tickborne diseases among this demographic. This study aimed to determine the knowledge and awareness among Illinois farmers regarding ticks and tickborne diseases. METHODS: A Knowledge, Attitudes & Prevention practices (KAP) survey was developed and administered to capture information regarding farmers' knowledge and attitudes about ticks and TBDs. Tick drags were conducted on a subset of properties as an incentive to complete the survey and to compare farmers' knowledge or expectations of ticks on their land with ticks collected. RESULTS: Fifty farmers participated in the survey, and 17 allowed tick drags. Only 60% of respondents had at least moderate knowledge about ticks gained through family and friends (56%), medical and healthcare personnel (48%), and the internet (44%). Responses varied by the type of commodity produced by the farmer. Fifty percent of participants reported knowledge about the blacklegged tick, 34% for the American dog tick, and 42% for the lone star tick; this knowledge also varied by farm type. Most farmers (54%) agreed that preventative behaviors could protect against tickborne diseases. Self-reported knowledge was significantly and directly associated with knowledge scores (p < .001). CONCLUSION: Knowledge of ticks and TBDs was lower among crop farmers than beef or mixed commodity farmers, but farmers generally have moderate knowledge of tick species in Illinois. Many participants expressed low concern over contracting a TBD, but many were also dissatisfied with the level of tick prevention measures that they follow. These results can be utilized to fill in knowledge gaps and develop informational materials to help farmers protect themselves against ticks and TBDs.

Knowledge, Attitudes, and Practices of Pastoralists Towards Tick Bites, and Tick Control in Plateau State, Nigeria.

PURPOSE: Pastoralists regularly come in contact with ticks as they herd their animals and are exposed to pathogens that cause zoonotic diseases. No study has been conducted in Nigeria to evaluate the knowledge, attitudes, and practices (KAP) of these Pastoralists towards ticks, tick bite, and tick control, and thus this research. METHODS: A KAP survey of pastoralists (n = 119) was conducted in Plateau State, Nigeria. Data generated were analysed using Statistical Package for Social Sciences (SPSS). RESULTS: The majority of the pastoralists (99.2%) had knowledge of ticks, with 79% of them being aware that ticks attach and bite humans, whereas only 30.3% believed that ticks transmit diseases to humans. Eighty-four per cent of the pastoralists do not wear protective clothing while herding their animals and 81.5% indicated to having been bitten by ticks, whereas hospital visit after tick bite was low (7.6%). Statistically significant variables were observed when knowledge of the respondents were compared in relation to the ability of ticks to cause diseases (Χ2 = 9.980, P = 0.007); hospital visit after a bite (Χ2 = 11.453, P = 0.003); and the use of protective clothing for herding (Χ2 = 22.596, P = 0). The main tick control measure was hand picking (58.8%). CONCLUSIONS: The pastoralists were unaware of the ability of ticks to transmit zoonotic pathogens. Preventive practices were insufficient to reduce tick bites, and thus were constantly exposed to tick-borne diseases. This study hopes to provide important insights for the development of educational awareness programmes for the pastoralists and serve as a guide for the health workers in designing future preventive programmes against tick-borne zoonoses in Nigeria.

Biodiversity in the Lyme-light: ecological restoration and tick-borne diseases in Europe.

Biodiversity loss and the emergence of zoonotic diseases are two major global challenges. An urgent question is how ecosystems and wildlife communities can be restored whilst minimizing the risk of zoonotic diseases carried by wildlife. Here, we evaluate how current ambitions to restore Europe's natural ecosystems may affect the hazard of diseases vectored by the tick Ixodes ricinus at different scales. We find that effects of restoration efforts on tick abundance are relatively straightforward but that the interacting effects of vertebrate diversity and abundance on pathogen transmission are insufficiently known. Long-term integrated surveillance of wildlife communities, ticks, and their pathogens is needed to understand their interactions and to prevent nature restoration from increasing tick-borne disease (TBD) hazard.

Impacts Over Time of Neighborhood-Scale Interventions to Control Ticks and Tick-Borne Disease Incidence.

Background: Controlling populations of ticks with biological or chemical acaricides is often advocated as a means of reducing human exposure to tick-borne diseases. Reducing tick abundance is expected to decrease immediate risk of tick encounters and disrupt pathogen transmission cycles, potentially reducing future exposure risk. Materials and Methods: We designed a placebo-controlled, randomized multiyear study to assess whether two methods of controlling ticks-tick control system (TCS) bait boxes and Met52 spray-reduced tick abundance, tick encounters with people and outdoor pets, and reported cases of tick-borne diseases. The study was conducted in 24 residential neighborhoods in a Lyme disease endemic zone in New York State. We tested the hypotheses that TCS bait boxes and Met52, alone or together, would be associated with increasing reductions in tick abundance, tick encounters, and cases of tick-borne disease over the 4-5 years of the study. Results: In neighborhoods with active TCS bait boxes, populations of blacklegged ticks (Ixodes scapularis) were not reduced over time in any of the three habitat types tested (forest, lawn, shrub/garden). There was no significant effect of Met52 on tick abundance overall, and there was no evidence for a compounding effect over time. Similarly, we observed no significant effect of either of the two tick control methods, used singly or together, on tick encounters or on reported cases of tick-borne diseases in humans overall, and there was no compounding effect over time. Thus, our hypothesis that effects of interventions would accumulate through time was not supported. Conclusions: The apparent inability of the selected tick control methods to reduce risk and incidence of tick-borne diseases after years of use requires further consideration.

Knowledge, attitudes, and practices for tick bite prevention and tick control among residents of Long Island, New York, USA.

In the United States, tick-borne diseases (TBDs) are a major public health concern and a source of significant morbidity. To reduce tick-borne disease incidence, understanding factors associated with increased risk in humans for tick bites is necessary. These risk factors can include TBD knowledge, attitudes about prevention and care, and associated practices of individuals and their communities, including paying for preventive services. Our study focused on Long Island, New York, a region with high endemicity of ticks and TBDs. The purpose of our study was to identify gaps in the knowledge, attitudes, and practices regarding TBDs in residents of this region to inform priorities for TBD interventions. To this end, we performed a knowledge, attitudes, and practices (KAP) survey and collected 803 responses from Long Island residents. Our survey results demonstrated that Long Island residents had a low to moderate level of knowledge regarding ticks and TBDs. Still, residents expressed concern regarding TBDs, with increased levels of concern associated with increased likelihood of paying for tick control and for practicing tick bite prevention strategies. Individuals with pets were more likely to practice these tick bite prevention strategies. Residents with more frequent exposure to ticks, with a history of TBD in their household, and with higher general concern over ticks were also more likely to pay for tick control services. These findings highlight gaps in resident knowledge, differences in attitudes towards tick preventative behavior, and potential factors affecting motivation towards implementing tick control measures, which can inform future public health messaging regarding tick bite prevention.

Tick-Borne Diseases.

Many tick-borne infections are increasing in geographic range as a result of activities such as reforestation and climate change. A history of outdoor activity in tick-endemic regions, together with consistent clinical signs (such as fever, splenomegaly, polyarthritis, thrombocytopenia), should raise suspicion for tick-borne infectious disease. Diseases with short incubation periods are best diagnosed with organism-detection assays. When the incubation period is long and organism numbers are low, diagnosis often relies on antibody testing, but it may be difficult to associate positive tests with infection. Positive antibody tests in healthy animals should prompt veterinarians to discuss prevention approaches with owners.