Molecular analysis of human tick-bitten skin yields signatures associated with distinct spatial and temporal trajectories - A proof-of-concept study.

Tick-associated diseases present challenges due to tridirectional interactions among host-specific responses, tick toxins and salivary proteins as well as microbes. We aimed to uncover molecular mechanisms in tick-bitten skin samples (cases) and contralateral skin samples (controls) collected simultaneously from the same participants, using spatial transcriptomics. Cases and controls analysed using NanoString GeoMx Digital Spatial Profiler identified 274 upregulated and 840 downregulated differentially expressed genes (DEGs), revealing perturbations in keratinization and immune system regulation. Samples of skin biopsies taken within 72 h post tick-bite DEGs had changes in protein metabolism and viral infection pathways as compared to samples taken 3 months post tick-bite, which instead displayed significant perturbations in several epigenetic regulatory pathways, highlighting the temporal nature of the host response following tick bites. Within-individual signatures distinguished tick-bitten samples from controls and identified between-individual signatures, offering promise for future biomarker discovery to guide prognosis and therapy.

Clinical Study and Serological Diagnosis of Vector-Borne Pathogens in Sardinian Dogs.

Canine vector-borne diseases (CVBDs) comprise a group of infectious diseases caused by a wide range of pathogens transmitted by arthropod vectors. Clinical signs commonly involve symptoms such as fever, anorexia, weight loss, blood disorders, hepatosplenomegaly, and others that can lead to death in dogs with comorbidities. Some pathogens responsible for CVBDs constitute a serious threat to human health due to their zoonotic transmission. This study aimed to determine the prevalence of zoonotic vector-borne diseases (Rickettsia rickettsii, Anaplasma phagocytophilum, Ehrlichia canis, Bartonella henselae, and Leishmania infantum) in domestic Sardinian dogs with and without clinical signs of these pathogens. Blood serum samples were collected from 142 dogs and examined through serological analysis. Clinical signs suggestive of these pathogens were also evaluated. The results obtained showed that 33 (33/140; 23.6%), 22 (22/134; 16.4%), 14 (14/142; 9.9%), 20 (20/66; 30.3%), and 26 (26/108; 24.1%) dogs were seropositive for Rickettsia sp., Anaplasma sp., Ehrlichia sp., Bartonella sp., and Leishmania sp. antibodies, respectively. Among these dogs, 12 dogs presented with at least one clinical sign (8.5%), while 18 (12.7%) showed more than two symptoms at the same time. Furthermore, among the asymptomatic dogs (93/142; 65.5%), 13% (n = 12) tested positive for A. phagocytophilum, 12% (n = 11) tested positive for B. henselae, 9% (n = 8) tested positive for E. canis, 12% (n = 11) tested positive for L. infantum, and 19% (n = 18) tested positive for R. rickettsii. This survey represents the first study assessing different canine vector-borne pathogens in dogs from North Sardinia. Since the pathogens detected here represent emerging zoonotic diseases, these results highlight the need to undertake further studies to increase the knowledge of these under-reported vector-borne pathogens in Sardinia.

Potential of mRNA-based vaccines for the control of tick-borne pathogens in one health perspective.

The One Health approach, which integrates the health of humans, animals, plants, and ecosystems at various levels, is crucial for addressing interconnected health threats. This is complemented by the advent of mRNA vaccines, which have revolutionized disease prevention. They offer broad-spectrum effectiveness and can be rapidly customized to target specific pathogens. Their utility extends beyond human medicine, showing potential in veterinary practices to control diseases and reduce the risk of zoonotic transmissions. This review place mRNA vaccines and One Health in the context of tick-borne diseases. The potential of these vaccines to confer cross-species immunity is significant, potentially disrupting zoonotic disease transmission cycles and protecting the health of both humans and animals, while reducing tick populations, infestations and circulation of pathogens. The development and application of mRNA vaccines for tick and tick-borne pathogens represent a comprehensive strategy in global health, fostering a healthier ecosystem for all species in our interconnected world.

Epidemiology and ecology of the sylvatic cycle of African Swine Fever Virus in Kenya.

African Swine Fever (ASF) is caused by a DNA virus (AFSV) maintained and transmitted by the Argasid ticks. The re-emergence of the disease in Africa coupled with its rapid spread globally is a threat to the pig industry, food security and livelihoods. The ecology and epidemiology of the ASFV sylvatic cycle, especially in the face of changing land use and land cover, further compounds the menace and impacts of this disease in Kenya. The study aimed to determine the occurrence and distribution of ASFV seroprevalence in warthog populations, the tick vectors and extent of tick infestation of warthog burrows, and the genotypes of ASFV in soft ticks in Kenya. Warthogs from different parts of Kenya were captured and venous blood was centrifuged to harvest sera. Warthog burrows were examined for their conditions and to extract ticks. Sera were analyzed for antibodies against ASFV using a commercial ELISA kit coated with p32 ASFV recombinant protein. Ticks were pooled, DNA extracted and the p72 gene of the ASFV was amplified by qPCR and conventional PCR. The overall seroprevalence of ASFV in warthogs was 87.5 %. A total of 228 warthog burrows were examined and 2154 argasid ticks were extracted from the burrows. Tick pools from Kigio Farm and Lewa Wildlife Conservancies were ASFV-positive by qPCR and conventional PCR. ASFV was further confirmed by the Twist Comprehensive Viral Research Panel (TCVRP), which also identified the argasid ticks as Ornithodoros porcinus. The ticks were infected with virus genotype IX, and their occurrence overlaps with regions of previous ASF outbreaks in domestic pigs. Further, Viruses that could be tick endosymbionts/commensals or due to bloodmeal were detected in ticks by TCVRP; Porcine type-C oncovirus; Pandoravirus neocaledonia; Choristoneura fumiferana granulovirus; Enterobacteria phage p7; Leporid herpesvirus 4 isolate; 5; Human Lymphotropic virus; Human herpesvirus 5. In conclusion, our results suggest that infected Ornithodoros spp. seems to have a rich virome, which has not been explored but could be exploited to inform ASF control in Kenya. Further, the ecology of Ornithodoros spp. and burrow-use dynamics are complex and more studies are needed to understand these dynamics, specifically in the spread of ASFV at the interface of wild and domestic pigs. Further, our results provide evidence of genotype IX ASFV sylvatic cycle which through O. porcinus tick transmission has resulted in high exposure of adult common warthogs. Finally, the co-circulation of ASFV genotype IX in the same location with past ASF outbreaks in domestic pigs and presently in ticks brings to focus the role of the interface and ticks on virus transmission to pigs and warthogs.

A Set of Diagnostic Tests for Detection of Active Babesia duncani Infection.

OBJECTIVES: Human babesiosis, an emerging potential fatal tick-borne disease caused by intraerythrocytic parasites of the Babesia genus. B. duncani is one of the Babesia species cause severe and life-threatening infections in humans. Detecting B. duncani infection is essential for accurate diagnosis and effective disease management. While molecular assays for detection in blood exist, there is still no reliable method to detect biomarkers of active infection. METHODS: We developed the first B. duncani antigen capture assays, targeting two immunodominant antigens, BdV234 and BdV38. These assays were validated using established in vitro and in vivo B. duncani-infection levels, both before and after therapy. RESULTS: The assay displayed no cross-reactivity with other species such as B. microti, B. divergens, Babesia MO1, or P. falciparum. It can detect as few as 115 infected erythrocytes/µl blood. Screening of 1,731 blood samples from various biorepositories, including samples previously identified as Lyme and/or B. microti positive, as well as new specimens from field mice, revealed no evidence of B. duncani infection and cross reactivity. CONCLUSION: These assays have potential applications, such as point-of-care testing for early detection of B. duncani in patients, field tests for screening reservoir hosts, and high-throughput screening of blood samples collected for transfusion.

Molecular detection of Coxiella burnetii in ticks collected from animals and the environment in Uganda.

AIMS: Coxiella burnetii is a highly infectious organism that is easily spread through aerosols causing Q fever in humans. Ticks can harbour and transmit C. burnetii to animals, contributing to disease maintenance. Our aim was to examine the presence of C. burnetii in ticks in Uganda. METHODS AND RESULTS: In this study, ticks were collected from five Ugandan districts and tested by real-time PCR for C. burnetii (Coxiella outer membrane protein 1 gene). A total of 859 tick pools (9602 individual ticks) were tested, and pool positivity for C. burnetii was 5.5% (n = 47). Pooled prevalence differed by district; the highest was Luwero (7.3%), then Gulu (6.6%), and Kasese had the lowest (1.3%). However, district variation was not statistically significant (Fisher's exact = 0.07). Ticks collected from dogs and cats had the highest positivity rates [23/47, (48.9%)] followed by livestock (cattle, goats, sheep, and pigs) [18/47, (38.3%)] and vegetation [6/47, (12.8%)]. Haemaphysalis elliptica had the highest infection rates, followed by Rhipicephalus appendiculatus, Amblyomma variegatum and Rhipicephalus decoloratus had similar prevalence. CONCLUSIONS: Although ticks are not the primary transmitters of C. burnetii to humans, pathogen detection in ticks can be an indirect indicator of risk among animal hosts. Vulnerable populations, including occupations with close animal contact such as farming, butchery, and veterinary practice, have an increased risk of C. burnetii exposure. Veterinarians and clinicians should be aware that C. burnetii may cause human and animal illness in these regions.

A wide distribution of Beiji nairoviruses and related viruses in Ixodes ticks in Japan.

Beiji nairovirus (BJNV), in the family Nairoviridae, the order Bunyavirales, was recently reported as a causative agent of an emerging tick-borne zoonotic infection in China. This study investigated the prevalence of BJNV in ticks in Japan. Screening of over 2,000 ticks from multiple regions revealed a widespread distribution of BJNV and BJNV-related viruses in Japan, particularly in the northern island, and in other high altitude areas with exclusive occurrence of Ixodes ticks. Phylogenetic analysis identified three distinct groups of nairoviruses in ticks in Japan: BJNV, Yichun nairovirus (YCNV) and a newly identified Mikuni nairovirus (MKNV). BJNV and YCNV variants identified in ticks in Japan exhibited high nucleotide sequence identities to those in China and Russia with evidence of non-monophyletic evolution among BJNVs, suggesting multiple cross-border transmission events of BJNV between the Eurasian continent and Japan. Whole genome sequencing of BJNV and MKNV revealed a unique GA-rich region in the S segment, the significance of which remains to be determined. In conclusion, the present study has shown a wide distribution and diversity of BJNV-related nairoviruses in Ixodes ticks in Japan and has identified unique genomic structures. The findings demonstrate the significance of BJNV as well as related viruses in Japan and highlight the necessity of monitoring emerging nairovirus infections and their potential risks to public health.

Assessing the value and knowledge gains from an online tick identification and tick-borne disease management course for the Southeastern United States.

BACKGROUND: Tick-borne diseases are a growing public health threat in the United States. Despite the prevalence and rising burden of tick-borne diseases, there are major gaps in baseline knowledge and surveillance efforts for tick vectors, even among vector control districts and public health agencies. To address this issue, an online tick training course (OTTC) was developed through the Southeastern Center of Excellence in Vector-Borne Diseases (SECOEVBD) to provide a comprehensive knowledge base on ticks, tick-borne diseases, and their management. METHODS: The OTTC consisted of training modules covering topics including tick biology, tick identification, tick-borne diseases, and public health, personal tick safety, and tick surveillance. The course was largely promoted to vector control specialists and public health employees throughout the Southeastern US. We collected assessment and survey data on participants to gauge learning outcomes, perceptions of the utility of knowledge gained, and barriers and facilitators to applying the knowledge in the field. RESULTS: The OTTC was successful in increasing participants' baseline knowledge across all course subject areas, with the average score on assessment increasing from 62.6% (pre-course) to 86.7% (post-course). More than half of participants (63.6%) indicated that they would definitely use information from the course in their work. Barriers to using information identified in the delayed assessment included lack of opportunities to apply skills (18.5%) and the need for additional specialized training beyond what the OTTC currently offers (18.5%), while the main facilitator (70.4%) for applying knowledge was having opportunities at work, such as an existing tick surveillance program. CONCLUSIONS: Overall, this OTTC demonstrated capacity to improve knowledge in a necessary and underserved public health field, and more than half of participants use or plan to use the information in their work. The geographic reach of this online resource was much larger than simply for the Southeastern region for which it was designed, suggesting a much broader need for this resource. Understanding the utility and penetrance of training programs such as these is important for refining materials and assessing optimal targets for training.

Evolution of Lyme Borreliosis Epidemiology in Primary Care and Hospital Settings in France During the COVID-19 Pandemic.

Background: Lyme borreliosis (LB) remains a public health concern in France despite improved patient management and medical care. Stay-at-home restrictions during the COVID-19 pandemic, which affected participation in outdoor recreational activities and disrupted access to health care services, may have impacted the risk of developing LB. Methods: We analyzed data from two general practitioner networks in France (Sentinel Network and an electronic medical records database [EMR]) and the national hospital discharge database to describe LB epidemiology in 2020-2021 and compare it to previous years. Google Trends' search volume was used to evaluate the association between the population's interest in LB and the evolving epidemiology. Results: Annual LB incidence rates in primary care decreased from 104 cases/100,000 population in 2018 to 71/100,000 in 2021 and from 82/100,000 to 60/100,000 according to Sentinel Network and EMR, respectively. Google Trends' search volume for "Lyme" followed a similar trend, one year earlier. Annual hospitalizations were stable from 2012-2019 (1.6/100,000 on average) and declined to 1.3/100,000 in 2020 and 1.1/100,000 in 2021. This decline was observed primarily in adults (e.g., 3.4/100,000 in 2017-2019 to 1.8/100,000 in 2020-2021 for 70-79 years of age). Changes in regional incidence rates in primary care from 2017-2019 to 2020-2021 ranged from -75% to 208%. Hospitalizations decreased in all regions except in Bretagne. Conclusions: The estimated LB incidence decreased in 2020 and 2021 compared with previous years but this change may not be related to COVID-19. The incidence decrease observed in primary care could result from reduced population interest in LB, leading to lower care-seeking behavior. The decrease in LB hospitalizations may be explained by changes in clinical practice. Surveillance systems are critical to understand the evolution of LB epidemiology. However, external factors impacting incidence estimates should be considered.

Endemisation and management of Babesia divergens on a beef production farm.

The hard tick Ixodes ricinus transmits a variety of zoonotic pathogens, including Babesia divergens, the most common cause of bovine babesiosis in northern Europe. In endemic areas, cattle are rarely clinically affected, as animals up to the age of nine months are resistant against relevant clinical disease and develop protective premunity. However, outbreaks in immunologically naïve herds may lead to considerable losses. Such an outbreak with a high mortality rate occurred in 2018 on a northern German beef production farm, as previously reported. The present study provides an update on the epidemiological situation and management strategy of the farm. In spring 2022, blood samples were taken from 46 animals for PCR and serological testing before pasture turnout. Although no clinical cases had been noticed since 2019, B. divergens DNA was detected by quantitative real-time PCR (qPCR), followed by amplification and sequencing of the 18S rRNA gene, in 6.5% (3/46) of cattle blood samples. Presence of anti-B. divergens antibodies was confirmed in 26.1% (12/46) of animals, while further 10.9% (5/46) had a borderline antibody titre. The antibody status of 23 of these animals had already been determined in 2018 and/or 2020, revealing fluctuating titre patterns indicative of repeated pathogen exposure. Moreover, 457 questing I. ricinus specimens collected on the farm's pastures and 83 I. ricinus specimens detached from cattle were screened for Babesia spp. DNA by qPCR, followed by 18S rDNA amplification and sequencing. Endemisation of B. divergens was confirmed by 0.9% (4/457) positive questing I. ricinus, while the ticks detached from cattle were Babesia-negative. The farm's management strategy includes annual metaphylactic treatment with imidocarb dipropionate during the main tick exposure period in spring. However, the antibody titre fluctuations and the persistent infections at the end of the housing period indicate that the absence of clinical disease is primarily due to a rising level of premunity. Metaphylactic treatment with imidocarb seems to be a suitable management option to protect newly acquired immunologically naïve animals. The endemisation of B. divergens is also of public health significance, as the pastures are located close to a tourist destination in a popular hiking area.

The role of wildlife in the epidemiology of tick-borne diseases in Slovakia.

Tick-borne diseases (TBD) represent an important challenge for human and veterinary medicine. In Slovakia, studies on the epidemiology of tick-borne pathogens (TBP) regarding reservoir hosts have focused on small mammals and to a lesser extent on birds or lizards, while knowledge of the role of the remaining vertebrate groups is limited. Generally, wild ungulates, hedgehogs, small- and medium-sized carnivores, or squirrels are important feeding hosts for ticks and serve as reservoirs for TBP. Importantly, because they carry infected ticks and/or are serologically positive, they can be used as sentinels to monitor the presence of ticks and TBP in the environment. With their increasing occurrence in urban and suburban habitats, wild ungulates, hedgehogs or foxes are becoming an important component in the developmental cycle of Ixodes ricinus and of TBP such as Anaplasma phagocytophilum or Babesia spp. On the other hand, it has been postulated that cervids may act as dilution hosts for Borrelia burgdorferi (sensu lato) and tick-borne encephalitis virus. In southwestern Slovakia, a high prevalence of infection with Theileria spp. (100%) was observed in some cervid populations, while A. phagocytophilum (prevalence of c.50%) was detected in cervids and wild boars. The following pathogens were detected in ticks feeding on free-ranging ungulates, birds, and hedgehogs: A. phagocytophilum, Rickettsia spp., Coxiella burnetii, Neoehrlichia mikurensis, B. burgdorferi (s.l.), and Babesia spp. The growing understanding of the role of wildlife as pathogen reservoirs and carriers of pathogen-infected ticks offers valuable insights into the epidemiology of TBP, providing a foundation for reducing the risk of TBD.

Germany-wide citizen science study reveals spread of Babesia canis-infected Dermacentor reticulatus ticks by dogs travelling within the country.

The ornate dog tick Dermacentor reticulatus, vector of Babesia canis, has shown a considerable range expansion in several European countries. Previously, only few areas in Germany were recognised as endemic for B. canis, but a marked increase in autochthonous canine babesiosis cases and spread to new areas has been noted recently. To better assess the current risk for dogs, the present study screened 5913 specimens of D. reticulatus from all over Germany, collected in the frame of a Citizen Science study during 2019-2023. Moreover, 343 Dermacentor marginatus ticks were also included. Babesia detection was achieved by quantitative real-time PCR (qPCR). Positive samples were confirmed by sequencing. Moreover, a MGB-probe-based triplex qPCR was established to detect and distinguish between the canine Babesia spp. relevant in Europe, i.e. B. canis, Babesia vogeli and Babesia gibsoni. Overall, B. canis DNA was detected in five D. reticulatus specimens (0.08%). Two of the B. canis-positive ticks originated from areas previously known as endemic for canine babesiosis, namely from the area of Freiburg im Breisgau, federal state of Baden-Wuerttemberg, and from the district St. Wendel, federal state of Saarland. Three further B. canis-positive ticks were detected in districts not yet recognised as endemic, one each in the district of Mansfeld-Suedharz, federal state of Saxony-Anhalt, the district of Ravensburg, federal state of Baden-Wuerttemberg and in the city of Fürth, federal state of Bavaria. However, the tick in Fürth was found on a dog who had returned from a trip to the Breisgau region on the previous day, indicating translocation of the specimen out of this well-known endemic focus. The geographical distribution of the positive samples shows that B. canis is currently spreading in Germany, particularly via dogs travelling within the country, increasing the infection risk throughout the country. Important measures to contain a further spread of the pathogen include comprehensive year-round tick prophylaxis with licensed acaricides, not only to protect the individual pet, but also the entire dog population. Moreover, screening of dogs entering Germany from B. canis-endemic countries is required and any treatment should aim at pathogen elimination by use of appropriate imidocarb dosages.

Infectious risks associated with outdoor sports activities.

In France, outdoor sports and activities account for 36% of sports engagement, making outdoor venues the most popular settings for sports participation. Discussing the links between sports and health almost always highlights the beneficial impact of engaging in sports. However, due to a lack of specific notifications, infectious risks are not subject to epidemiological monitoring, and need to be better understood. Since the practice of outdoor sports has become part and parcel of many individuals' daily routines, it is essential to more accurately characterize the knowledge we have gained about the risks associated with exposure. However, directly associating the practice of a sport with an elevated risk of infectious diseases is a challenging endeavor. Sociological factors based on risk awareness and adoption of protective behaviors in response to the risk are crucial to the orientation of prevention efforts. This review deals with several (bacteriological, viral, parasitic, and mycological) infectious risks related to outdoor activities practiced in a natural field via contamination routes such as tick-bite, enteric pathogen, skin, and aerosol transmission. We have also detailed a number of preventive measures taking into account the outdoor setting (e.g., vaccination).

Seroprevalence study in humans and molecular detection in Rhipicephalus sanguineus ticks of severe fever with thrombocytopenia syndrome virus in Thailand.

Severe fever with thrombocytopenia syndrome virus (SFTSV) is an emerging tick-borne virus with a mortality rate of up to 30%. First identified in China in 2009, it was later reported in other Asian countries, including Thailand in 2020. SFTSV has been detected in several tick species, including Rhipicephalus sanguineus, known for infesting dogs. We conducted a seroprevalence study of SFTSV in Bangkok and Nong Khai, Thailand, by analyzing 1162 human samples collected between 2019 and 2023. The testing method relied on IgG detection using ELISA and confirmed though a virus seroneutralization test. The results indicated that out of the participants, 12 (1.1%) tested positive for anti-SFTSV IgG antibodies; however, none exhibited positive results in the seroneutralization assay. Additionally, molecular detection of SFTSV, Crimean-Congo hemorrhagic fever (CCHF), Coxiella spp., Bartonella spp., and Rickettsia spp. was performed on 433 Rh. sanguineus ticks collected from 49 dogs in 2023 in Chachoengsao Province, Thailand. No evidence of these pathogens was found in ticks. These findings highlight the importance of exploring viral cross-reactivity. Furthermore, it is important to conduct additional studies to isolate SFTSV from animals and ticks in order to identify the potential transmission routes contributing to human and animal infections in Thailand.

Molecular Detection of Anaplasma marginale in Capybara (Hydrochoerus hydrochaeris) from Corrientes, Argentina.

Monitoring wildlife health is essential for understanding global disease patterns, particularly as vector-borne infections extend the geographic ranges and thereby hosts due to environmental shifts. Anaplasma marginale, primarily impacting cattle, has economic implications and has been found in diverse hosts, yet its presence in capybaras (Hydrochoerus hydrochaeris), influential in tick-borne pathogen spread, lacks comprehensive understanding. From 2015 to 2022, 14 capybaras were surveyed across two different areas of northeastern Argentina. In 1 of 14 (7%) capybaras, the presence of A. marginale was confirmed through the amplification of specific genes, msp5 and msp1ß. In addition, A. marginale DNA was detected in the capybara's blood sample through quantitative PCR, with a cycle threshold value of 30.81 (800 copies per reaction). Amplification of a fragment of the msp1α gene revealed PCR products of three different sizes, suggesting the presence of at least three coinfecting A. marginale variants in the capybara host. This study suggests that capybaras are wild hosts for A. marginale in the Ibera Wetlands in Argentina, potentially influencing the infection dynamics of both domestic and wild species. This finding highlights the necessity for thorough studies on the role of capybaras in disease dynamics, crucial for understanding wildlife health and the spread of disease.

Molecular detection of rickettsial agents in Amblyomma maculatum ticks (Ixodida: Ixodidae) from Ecuador.

Rickettsiales are obligate intracellular bacteria that need vertebrates and arthropods to maintain their life cycles. Some species of the genera Anaplasma, Ehrlichia, and Rickettsia are transmitted by ticks to both animals and humans and can cause mild to severe and even fatal cases. In the Americas, there is substantial data on rickettsial agents, encompassing both clinical cases and the detection of these agents in ticks, but in Ecuador, the information about them remains poorly understood. Therefore, the objective of this study was to detect molecularly rickettsial agents in Amblyomma maculatum ticks in both parasitic and free-living phases collected from domestic animals and pasture in five localities across three coastal provinces of Ecuador. Rickettsia parkeri, Candidatus Rickettsia andeanae, and Ehrlichia sp. were recorded in A. maculatum for the first time in Ecuador. These records were made in a region where antibodies to the Spotted Fever Rickettsia Group were detected in humans. Additional studies are needed to characterize Ehrlichia sp. at a specific level. Furthermore, recognizing the specific Rickettsiales species circulating in the ticks and the hosts within a region is crucial for assessing potential contact risks.

Lime for Lyme: Treatment of Leaf Litter with Dolomitic Lime Powder Impairs Activity of Immature Ixodes scapularis Ticks.

Background: Tick-borne diseases are an emerging threat to public health throughout the temperate world, leading to a growing field of research aimed at developing and testing intervention strategies for reducing human-tick encounters or prevalence of infection in ticks. Various wide-spectrum chemical acaricides have proven effective for controlling tick populations, but many of these have potential deleterious side-effects on health and the environment. In addition to chemical acaricides, certain compounds such as diatomaceous earth have been shown to have physical acaricidal properties. We hypothesized that dolomitic lime (CaMg(CO3)2, a corrosive, desiccant mineral that is already used extensively in agricultural and forestry contexts to balance the pH of soils, may affect ticks' locomotory activity, habitat position, or survival and that this should manifest as a reduction in the number of questing ticks collected by dragging. Objective: This study aimed to formally assess this hypothesis in a controlled laboratory setting. Methods: We carried out a microcosm experiment, with one control and three treated microcosm trays, each replicating the natural substrate characterizing I. scapularis habitat in northeastern North America. Each tray was infested with 200 living larvae and 50 nymphs, and then treated with 0 (control), 50, 100, or 500 g/m2 of lime powder. Ticks were collected by microdragging 24 and 72 h postliming. Results: Efficacy of liming at reducing the number of collected questing ticks ranged from 87% to 100% for larvae and 0% to 69% for nymphs 24 h postliming and from 91% to 93% for larvae and -47% to 65% for nymphs 72 postliming. Conclusion: This study provides the first experimental evidence of the potential efficacy of liming for impairing activity of questing immature ticks. Given that lime is a low-cost material, that methods for widespread application in deciduous woodlands already exist, and that it has been documented as having a limited negative impact on the environment, further assessment of lime application as a public health risk reduction intervention for tick-borne diseases is warranted.

Tick-borne zoonotic flaviviruses and Borrelia infections in wildlife hosts: What have field studies contributed?

Tick-borne flaviviruses and Borrelia spp. are globally spread pathogens of zoonotic potential that are maintained by a transmission cycle at the interface between ticks and vertebrate hosts, mainly wild animals. Aside data on pathogen burden in ticks, information on the status of various hosts relative to infection is important to acquire. We reviewed how those infections have been studied in wildlife host species in the field to discuss how collected data provided relevant epidemiological information and to identify needs for further studies. The literature was screened for observational studies on pathogen or antibody detection for tick-borne Borrelia spp. and flaviviruses in wildlife host animals. Overall, Borrelia spp. were more studied (73% of case studies, representing 297 host species) than flaviviruses (27% of case studies, representing 114 host species). Studies on both Borrelia spp. and flaviviruses focused mainly on the same species, namely bank vole and yellow-necked mouse. Most studies were order-specific and cross-sectional, reporting prevalence at various locations, but with little insight into the underlying epidemiological dynamics. Host species with potential to act as reservoir hosts of these pathogens were neglected, notably birds. We highlight the necessity of collecting both demographics and infection data in wildlife studies, and to consider communities of species, to better estimate zoonotic risk potential in the One Health context.

Widespread Circulation of Tick-Borne Viruses in Virginia-Evidence of Exposure to Heartland, Bourbon, and Powassan Viruses in Wildlife and Livestock.

Emerging tick-borne viruses such as Powassan virus (POWV), Bourbon virus (BRBV), and Heartland virus (HRTV), whilst rare, can cause severe health problems in humans. While limited clinical cases have been reported thus far in Virginia, the presence of tick-borne viruses poses a serious health threat, and the extent of their prevalence in Virginia is unknown. Here, we sought evidence of POWV, BRBV, and HRTV exposure in Virginia via a serological assessment of wildlife and livestock. Wildlife in Virginia were found to be seropositive against POWV (18%), BRBV (8%), and HRTV (5%), with western and northern regions of the state having a higher prevalence. Multiple wildlife species were shown to have been exposed to each virus examined. To a lesser extent, cattle also showed exposure to tick-borne viruses, with seroprevalences of 1%, 1.2%, and 8% detected in cattle against POWV, BRBV, and HRTV, respectively. Cross-reactivity against other known circulating mosquito-borne flaviviruses was ruled out. In conclusion, there is widespread exposure to tick-borne viruses in western and northern Virginia, with exposure to a diverse range of animal populations. Our study provides the first confirmation that HRTV is circulating in the Commonwealth. These findings strengthen the existing evidence of emerging tick-borne viruses in Virginia and highlight the need for public health vigilance to avoid tick bites.

Novel Tick-Borne Anaplasmataceae Genotypes in Tropical Birds from the Brazilian Pantanal Wetland.

Despite numerous reports of Anaplasmataceae agents in mammals worldwide, few studies have investigated their occurrence in birds. The present study aimed to investigate the occurrence and molecular identity of Anaplasmataceae agents in birds from the Pantanal wetland, Brazil. Blood samples were collected from 93 different species. After DNA extraction, samples positive for the avian ß-actin gene were subjected to both a multiplex quantitative real-time (q)PCR for Anaplasma and Ehrlichia targeting the groEL gene and to a conventional PCR for Anaplasmataceae agents targeting the 16S rRNA gene. As a result, 37 (7.4%) birds were positive for Anaplasma spp. and 4 (0.8%) for Ehrlichia spp. in the qPCR assay; additionally, 13 (2.6%) were positive for Anaplasmataceae agents in the PCR targeting the 16S rRNA gene. The Ehrlichia 16S rRNA sequences detected in Arundinicola leucocephala, Ramphocelus carbo, and Elaenia albiceps were positioned closely to Ehrlichia sp. Magellanica. Ehrlichia dsb sequences detected in Agelasticus cyanopus and Basileuterus flaveolus grouped with Ehrlichia minasensis. The 16S rRNA genotypes detected in Crax fasciolata, Pitangus sulphuratus and Furnarius leucopus grouped with Candidatus Allocryptoplasma. The 23S-5S genotypes detected in C. fasciolata, Basileuterus flaveolus, and Saltator coerulescens were related to Anaplasma phagocytophilum. In conclusion, novel genotypes of Anaplasma, Ehrlichia, and Candidatus Allocryptoplasma were detected in birds from the Pantanal wetland.