The black-legged tick Ixodes scapularis detects CO2 without the Haller's organ.

Both male and female ticks have a strong innate drive to find and blood-feed on hosts. Carbon dioxide (CO2) is considered a critical behavioral activator and attractant for ticks and an essential sensory cue to find hosts. Yet, how CO2 activates and promotes host seeking in ticks is poorly understood. CO2 responses were studied in the black-legged tick Ixodes scapularis, the primary vector for Lyme disease in North America. Adult males and females were exposed to 1%, 2%, 4% or 8% CO2, and changes in walking behavior and foreleg movement were analyzed. CO2 is a potent stimulant for adult I. scapularis, even at lower concentrations (1%). Behavioral reactions depended on the animal's state: walking ticks increased their walking speed, while stationary ticks started to wave their forelegs and began to quest - both behaviors resembling aspects of host seeking. Only in sporadic cases did stationary animals start to walk when exposed to CO2, supporting the hypothesis that CO2 acts as an activator rather than an attractant. Furthermore, I. scapularis did not show a clear concentration preference and was not tuned more robustly to breath-like CO2 concentrations (∼4%) than to the other concentrations tested. Moreover, convincing evidence is provided showing that the foreleg Haller's organ is not necessary for CO2 detection. Even with a disabled or amputated Haller's organ, I. scapularis responded robustly to CO2, signifying that there must be CO2-sensitive structures important for tick host seeking that have not yet been identified.

Metabolomic changes associated with acquired resistance to Ixodes scapularis.

Guinea pigs repeatedly exposed to Ixodes scapularis develop acquired resistance to the ticks (ATR). The molecular mechanisms of ATR have not been fully elucidated, and partially involves immune responses to proteins in tick saliva. In this study, we examined the metabolome of sera of guinea pigs during the development of ATR. Induction of components of the tyrosine metabolic pathway, including hydroxyphenyllactic acid (HPLA), were associated with ATR. We therefore administered HPLA to mice, an animal that does not develop ATR, and exposed the animals to I. scapularis. We also administered nitisinone, a known inhibitor of tyrosine degradation, to another group of mice. The mortality of I. scapularis that fed on mice given HPLA or nitisinone was 26 % and 72 % respectively, compared with 2 % mortality among ticks that fed on control animals. These data indicate that tick bites alter the guinea pig metabolome, and that the tyrosine metabolism pathway can potentially be targeted for I. scapularis control.

Copulation of Ixodes ricinus males and females on the host and its potential impact on pathogen transmission.

INTRODUCTION AND OBJECTIVE: The common tick Ixodes ricinus is one of Europe's most important vectors of tick-borne diseases. The increased risk of attacks by this tick suggests the need for identification of factors contributing to the transmission of tick-borne pathogens, and the routes of pathogen circulation in nature. MATERIAL AND METHODS: Polymerase chain reaction was used to investigate the prevalence of four pathogens, i.e. Borrelia burgdorferi s.l. (Bb), Anaplasma phagocytophilum (Ap), Babesia spp. (Bs), and Bartonella spp. (Ba) in I. ricinus females and males mating on dogs in south-central Poland. RESULTS: The study revealed the presence of three pathogens: Bb, Ap, and Bs in 9.4%, 5.4%, and 5.4% of all I. ricinus adults in copula, respectively. Co-infection with two pathogens was detected in one tick specimen. Borrelia burgdorferi spirochetes were isolated in two females and two males in copula, but the sexual transfer of the spirochetes between these specimens could not be clearly confirmed. CONCLUSIONS: By increasing the feeding dynamics in females, the copulation of I. ricinus males with females attached to the host's skin may stimulate pathogen replication in tick tissues and migration from the gut to the salivary glands. Further investigations of the I. ricinus copulation on the host on female feeding and pathogen transmission may contribute to the elucidation of the eco-epidemiology of tick-borne diseases transmitted by this tick species.

Ixodes scapularis nymph saliva protein blocks host inflammation and complement-mediated killing of Lyme disease agent, Borrelia burgdorferi.

Tick serine protease inhibitors (serpins) play crucial roles in tick feeding and pathogen transmission. We demonstrate that Ixodes scapularis (Ixs) nymph tick saliva serpin (S) 41 (IxsS41), secreted by Borrelia burgdorferi (Bb)-infected ticks at high abundance, is involved in regulating tick evasion of host innate immunity and promoting host colonization by Bb. Recombinant (r) proteins were expressed in Pichia pastoris, and substrate hydrolysis assays were used to determine. Ex vivo (complement and hemostasis function related) and in vivo (paw edema and effect on Bb colonization of C3H/HeN mice organs) assays were conducted to validate function. We demonstrate that rIxsS41 inhibits chymase and cathepsin G, pro-inflammatory proteases that are released by mast cells and neutrophils, the first immune cells at the tick feeding site. Importantly, stoichiometry of inhibition analysis revealed that 2.2 and 2.8 molecules of rIxsS41 are needed to 100% inhibit 1 molecule of chymase and cathepsin G, respectively, suggesting that findings here are likely events at the tick feeding site. Furthermore, chymase-mediated paw edema, induced by the mast cell degranulator, compound 48/80 (C48/80), was blocked by rIxsS41. Likewise, rIxsS41 reduced membrane attack complex (MAC) deposition via the alternative and lectin complement activation pathways and dose-dependently protected Bb from complement killing. Additionally, co-inoculating C3H/HeN mice with Bb together with rIxsS41 or with a mixture (rIxsS41 and C48/80). Findings in this study suggest that IxsS41 markedly contributes to tick feeding and host colonization by Bb. Therefore, we conclude that IxsS41 is a potential candidate for an anti-tick vaccine to prevent transmission of the Lyme disease agent.

Differential associations of horizontally and vertically transmitted symbionts on Ixodes ricinus behaviour and physiology.

BACKGROUND: Ixodes ricinus ticks are infected with a large diversity of vertically and horizontally transmitted symbionts. While horizontally transmitted symbionts rely on a vertebrate host for their transmission, vertically transmitted symbionts rely more on the survival of their invertebrate host for transmission. We therefore hypothesized horizontally transmitted symbionts to be associated with increased tick activity to increase host contact rate and vertically transmitted symbionts to be associated with higher tick weight and lipid fraction to promote tick survival. METHODS: We used a behavioural assay to record the questing activity of I. ricinus ticks. In addition, we measured weight and lipid fraction and determined the presence of ten symbiont species in these ticks using qPCR, of which six were vertically transmitted and four horizontally transmitted. RESULTS: Vertically transmitted symbionts (e.g. Midichloria mitochondrii) were associated with an increase in tick weight, whereas horizontally transmitted symbionts (e.g. Borrelia burgdorferi sensu lato) were often associated with lower weight and lipid fraction of ticks. Moreover, horizontally transmitted symbionts (e.g. B. burgdorferi s.l.) were associated with increased tick activity, which may benefit pathogen transmission and increases tick-borne disease hazard. CONCLUSIONS: Our study shows that horizontally and vertically transmitted symbionts differentially influence the behaviour and physiology of I. ricinus and warrants future research to study the underlying mechanisms and effects on transmission dynamics of tick-borne pathogens.

Adiponectin in the mammalian host influences ticks' acquisition of the Lyme disease pathogen Borrelia.

Arthropod-borne pathogens cause some of the most important human and animal infectious diseases. Many vectors acquire or transmit pathogens through the process of blood feeding. Here, we report adiponectin, the most abundant adipocyte-derived hormone circulating in human blood, directly or indirectly inhibits acquisition of the Lyme disease agent, Borrelia burgdorferi, by Ixodes scapularis ticks. Rather than altering tick feeding or spirochete viability, adiponectin or its associated factors induces host histamine release when the tick feeds, which leads to vascular leakage, infiltration of neutrophils and macrophages, and inflammation at the bite site. Consistent with this, adiponectin-deficient mice have diminished pro-inflammatory responses, including interleukin (IL)-12 and IL-1ß, following a tick bite, compared with wild-type animals. All these factors mediated by adiponectin or associated factors influence B. burgdorferi survival at the tick bite site. These results suggest a host adipocyte-derived hormone modulates pathogen acquisition by a blood-feeding arthropod.

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

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

Interspecific differences in the behavioral response of ticks exposed to radiofrequency electromagnetic radiation.

Artificial electromagnetic radiation is a new environmental factor that affects animals. Experiments with the effect of radio frequency electromagnetic radiation were focused on both vertebrates and invertebrates. Ticks showed a significant affinity to radiation. Our study is a continuation of this research and its aim was to monitor the effect of radiation on the behavior of four tick species: Ixodes ricinus, Dermacentor reticulatus, Dermacentor marginatus and Haemaphysalis inermis. In total 1,200 ticks, 300 of each species, were tested in modules allowing the choice of an exposed or shielded area. During the test, the ticks were exposed to electro-magnetic radiation of 900 MHz for 24 h. The position of the individuals was recorded and we evaluated the obtained data statistically. We observed a significant preference to the exposed area in both sexes of I. ricinus. Males of D. reticulatus and D. marginatus also showed an affinity to radiation, but not females of both species, nor females and males of H. inermis. The results of the study support the assumption that ticks perceive the electromagnetic field and the observed differences in their response have the potential to help understand the mechanism of perception.

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

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

Microbiota perturbation by anti-microbiota vaccine reduces the colonization of Borrelia afzelii in Ixodes ricinus.

BACKGROUND: Ticks can transmit a broad variety of pathogens of medical importance, including Borrelia afzelii, the causative agent of Lyme borreliosis in Europe. Tick microbiota is an important factor modulating, not only vector physiology, but also the vector competence. Anti-microbiota vaccines targeting keystone taxa of tick microbiota can alter tick feeding and modulate the taxonomic and functional profiles of bacterial communities in the vector. However, the impact of anti-microbiota vaccine on tick-borne pathogen development within the vector has not been tested. RESULTS: Here, we characterized the Ixodes ricinus microbiota modulation in response to B. afzelii infection and found that the pathogen induces changes in the microbiota composition, its beta diversity and structure of bacterial community assembly. Tick microbiota perturbation by anti-microbiota antibodies or addition of novel commensal bacteria into tick midguts causes departures from the B. afzelii-induced modulation of tick microbiota which resulted in a lower load of the pathogen in I. ricinus. Co-occurrence networks allowed the identification of emergent properties of the bacterial communities which better defined the Borrelia infection-refractory states of the tick microbiota. CONCLUSIONS: These findings suggest that Borrelia is highly sensitive to tick microbiota perturbations and that departure from the modulation induced by the pathogen in the vector microbiota pose a high cost to the spirochete. Network analysis emerges as a suitable tool to identify emergent properties of the vector microbiota associated with infection-refractory states. Anti-microbiota vaccines can be used as a tool for microbiota perturbation and control of important vector-borne pathogens. Video Abstract.

Topical permethrin may increase blacklegged tick (Ixodes scapularis) repellency but is associated with cutaneous irritation in horses.

OBJECTIVE: To evaluate the safety of repeated applications of permethrin concentrations (0% control, 1.5%, 5%, and 10%) to the necks and faces of horses and assess the efficacy and longevity of permethrin as an equine tick repellent. ANIMALS: 5 healthy adult Quarter Horses. PROCEDURES: Each treatment was applied to the neck of each horse (0.01 m2) 4 times a day, for up to 10 days. An 8-mm biopsy was taken to evaluate postexposure dermal responses. Any treatments that were not withdrawn were applied to a quadrant of the horse's face 4 times a day, for up to 5 days. For tick bioassays, a treatment was applied to 1 leg of a horse and 5 female blacklegged ticks (Ixodes scapularis) were evaluated as "repelled" or "not repelled" by the treatment. The bioassays were repeated up to 5 days, but treatment application took place only on the first day of the experiment. RESULTS: Histological results of neck biopsies indicated that more repeated exposures or higher concentrations resulted in more dermal damage. Tick bioassays showed that 5% and 10% permethrin had the greatest efficacy and longevity as a tick repellent, but the differences in tick repellency were not significant overall. CLINICAL RELEVANCE: While there was a nonsignificant trend of higher permethrin concentrations repelling more ticks with longer-lasting residual repellent effects, higher concentrations also produced greater skin damage after repeated exposures. These opposing findings emphasize the need for better tick prevention and control methods that balance safety and efficacy for the equine community.

Tick saliva and its role in pathogen transmission.

Tick saliva is a complex mixture of peptidic and non-peptidic molecules that aid engorgement. The composition of tick saliva changes as feeding progresses and the tick counters the dynamic host response. Ixodid ticks such as Ixodes ricinus, the most important tick species in Europe, transmit numerous pathogens that cause debilitating diseases, e.g. Lyme borreliosis and tick-borne encephalitis. Tick-borne pathogens are transmitted in tick saliva during blood feeding; however, saliva is not simply a medium enabling pathogen transfer. Instead, tick-borne pathogens exploit saliva-induced modulation of host responses to promote their transmission and infection, so-called saliva-assisted transmission (SAT). Characterization of the saliva factors that facilitate SAT is an active area of current research. Besides providing new insights into how tick-borne pathogens survive in nature, the research is opening new avenues for vaccine development.

Out of Asia? Expansion of Eurasian Lyme borreliosis causing genospecies display unique evolutionary trajectories.

Vector-borne pathogens exist in obligate transmission cycles between vector and reservoir host species. Host and vector shifts can lead to geographic expansion of infectious agents and the emergence of new diseases in susceptible individuals. Three bacterial genospecies (Borrelia afzelii, Borrelia bavariensis, and Borrelia garinii) predominantly utilize two distinct tick species as vectors in Asia (Ixodes persulcatus) and Europe (Ixodes ricinus). Through these vectors, the bacteria can infect various vertebrate groups (e.g., rodents, birds) including humans where they cause Lyme borreliosis, the most common vector-borne disease in the Northern hemisphere. Yet, how and in which order the three Borrelia genospecies colonized each continent remains unclear including the evolutionary consequences of this geographic expansion. Here, by reconstructing the evolutionary history of 142 Eurasian isolates, we found evidence that the ancestors of each of the three genospecies probably have an Asian origin. Even so, each genospecies studied displayed a unique substructuring and evolutionary response to the colonization of Europe. The pattern of allele sharing between continents is consistent with the dispersal rate of the respective vertebrate hosts, supporting the concept that adaptation of Borrelia genospecies to the host is important for pathogen dispersal. Our results highlight that Eurasian Lyme borreliosis agents are all capable of geographic expansion with host association influencing their dispersal; further displaying the importance of host and vector association to the geographic expansion of vector-borne pathogens and potentially conditioning their capacity as emergent pathogens.

The added value of One Health surveillance: data from questing ticks can provide an early signal for anaplasmosis outbreaks in animals and humans.

OBJECTIVE: In 2021, a first outbreak of anaplasmosis occurred in animals and humans in southern Québec, with 64% of confirmed human cases located in Bromont municipality. Ixodes scapularis ticks and Peromyscus mouse ear biopsies collected in Bromont from 2019 to 2021 were analyzed for Anaplasma phagocytophilum (Ap) with the objective of determining whether an early environmental signal could have been detected before the outbreak. METHODS: Samples were collected for a concurrent study aiming to reduce Lyme disease risk. Between 2019 and 2021, up to 14 experimental sites were sampled for ticks and capture of small mammals took place on three sites in 2021. Samples were screened for Ap using multiplex real-time PCR, and genetic strains were identified using a single-nucleotide polymorphism assay. RESULTS: Analyses showed an increase of 5.7% in Ap prevalence in ticks (CI95: 1.5-9.9) between 2019 and 2020, i.e., one year before the outbreak. A majority of Ap-positive ticks were infected with the zoonotic strain (68.8%; CI95: 50.0-83.9) during the study period. In 2021, 2 of 59 captured Peromycus mice were positive for Ap, for a prevalence of 3.4% (CI95: 0.4-11.7). CONCLUSION: We conclude that data collected in Bromont could have provided an early signal for an anaplasmosis risk increasing in the targeted region. This is a reminder that integrated surveillance of tick-borne diseases through structured One Health programs, i.e. systematically integrating data from humans, animals and the environment, can provide useful and timely information for better preparedness and response in public health.

RéSUMé: OBJECTIF: En 2021, suivant une éclosion d'anaplasmoses chez les animaux et les humains dans le sud du Québec, des tiques de l'espèce Ixodes scapularis et des biopsies de souris Peromyscus spp. échantillonées à Bromont, la municipalité où 64 % des cas humains confirmés était localisé, ont été testées pour Anaplasma phagocytophilum (Ap) avec pour objectif de déterminer si un signal environnemental précoce d'augmentation du risque aurait pu être détecté avant l'éclosion. MéTHODE: L'échantillonnage a été réalisé dans le cadre d'une étude visant à réduire le risque de maladie de Lyme. De 2019 à 2021, 14 sites expérimentaux ont été échantillonnés pour les tiques. En 2021, trois sites ont été sélectionnés pour la capture des micromammifères. Les échantillons ont été testés pour la présence d'Ap à l'aide d'un PCR multiplex en temps réelle et les lignées génétiques ont été identifiées grâce à un test de polymorphisme mononucléotidique. RéSULTATS: Les analyses ont montré une augmentation de 5,7 % (IC95% : 1,5­9,9) de la prévalence de Ap entre 2019 et 2020, c'est-à-dire un an avant l'éclosion. Cette augmentation est associée à la présence d'une majorité d'Ap de la lignée zoonotique (68,8 %; IC95% : 50,0­83,9) sur l'ensemble de la période étudiée. En 2021, deux Peromycus spp. capturées sur 59 étaient positives pour Ap pour une prévalence de 3,4 % (IC95% : 0,4­11,7). CONCLUSION: Les données environnementales échantillonnées à Bromont auraient pu fournir un signal précoce de l'augmentation du risque d'anaplasmose dans la région. C'est un rappel que la surveillance intégrée des maladies transmises par les tiques inspirée de l'approche Une seule santé, intégrant systématiquement des données humaines, animales et environnementales, peut fournir des informations utiles et opportunes aux autorités de santé publique.

Interference with LTßR signaling by tick saliva facilitates transmission of Lyme disease spirochetes.

Lyme spirochetes have coevolved with ticks to optimize transmission to hosts using tick salivary molecules (TSMs) to counteract host defenses. TSMs modulate various molecular events at the tick-host interface. Lymphotoxin-beta receptor (LTßR) is a vital immune receptor and plays protective roles in host immunity against microbial infections. We found that Ltbr knockout mice were more susceptible to Lyme disease spirochetes, suggesting the involvement of LTßR signaling in tick-borne Borrelia infection. Further investigation showed that a 15-kDa TSM protein from Ixodes persulcatus (I. persulcatus salivary protein; IpSAP) functioned as an immunosuppressant to facilitate the transmission and infection of Lyme disease spirochetes. IpSAP directly interacts with LTßR to block its activation, thus inhibiting the downstream signaling and consequently suppressing immunity. IpSAP immunization provided mice with significant protection against I. persulcatus-mediated Borrelia garinii infection. Notably, the immunization showed considerable cross-protection against other Borrelia infections mediated by other ixodid ticks. One of the IpSAP homologs from other ixodid ticks showed similar effects on Lyme spirochete transmission. Together, our findings suggest that LTßR signaling plays an important role in blocking the transmission and pathogenesis of tick-borne Lyme disease spirochetes, and that IpSAP and its homologs are promising candidates for broad-spectrum vaccine development.

Tick feeding modulates the human skin immune landscape to facilitate tick-borne pathogen transmission.

During cutaneous tick attachment, the feeding cavity becomes a site of transmission for tick salivary compounds and tick-borne pathogens. However, the immunological consequences of tick feeding for human skin remain unclear. Here, we assessed human skin and blood samples upon tick bite and developed a human skin explant model mimicking Ixodes ricinus bites and tick-borne pathogen infection. Following tick attachment, we observed rapidly occurring patterns of immunomodulation, including increases in neutrophils and cutaneous B and T cells. T cells upregulated tissue residency markers, while lymphocytic cytokine production was impaired. In early stages of Borrelia burgdorferi model infections, we detected strain-specific immune responses and close spatial relationships between macrophages and spirochetes. Preincubation of spirochetes with tick salivary gland extracts hampered accumulation of immune cells and increased spirochete loads. Collectively, we showed that tick feeding exerts profound changes on the skin immune network that interfere with the primary response against tick-borne pathogens.

Role of bank vole (Myodes glareolus) personality on tick burden (Ixodes spp.).

Parasitism among individuals in a population often varies more than expected by chance only, leading to parasite aggregation, which is a parameter of paramount importance in parasite population dynamics and particularly in vector-borne epidemiology. However, the origin of this phenomenon is yet not fully elucidated. An increasing body of literature has demonstrated that individuals vary consistently in their behaviour, which is referred to as animal personality. Such behavioural variation could potentially lead to different encounter rates with parasites. To test this hypothesis, the relationship between personality and burden with ticks (Ixodes spp.) in the bank vole, Myodes glareolus (Schreber), was assessed. Wild rodents (eight females and 18 males) were live-trapped, identified, sexed, weighted, and inspected for ticks. Behavioural profiling was then performed using standardised tests measuring activity/exploration and boldness with a combination of automatically and manually recorded behavioural variables summarised using multivariate analyses. The resulting personality descriptors and questing tick variables were used as explanatory variables in negative binomial generalised linear models of tick burden and Bayesian simulations were performed to better estimate coefficients. Tick burden was associated to body mass and sex, but not to personality descriptors, which was mainly associated to activity/exploration. These results are discussed regarding the complex relationships among individual personality, physiological status, space use and health status.

Range Expansion of Ixodes scapularis and Borrelia burgdorferi in Ontario, Canada, from 2017 to 2019.

Range expansion of the vector tick species, Ixodes scapularis, has been detected in Ontario over the last two decades. This has led to elevated risk of exposure to Borrelia burgdorferi, the bacterium that causes Lyme disease. Previous research using passive surveillance data suggests that I. scapularis populations establish before the establishment of B. burgdorferi transmission cycles, with a delay of ∼5 years. The objectives of this research were to examine spatial and temporal patterns of I. scapularis and its pathogens from 2017 to 2019 in southwestern, eastern, and central Ontario, and to explore patterns of B. burgdorferi invasion. Over the 3-year study period, drag sampling was conducted at 48 sites across Ontario. I. scapularis ticks were tested for B. burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum, and Babesia species, including Babesia microti and Babesia odocoilei, and Powassan virus. I. scapularis was detected at 30 sites overall, 22 of which had no history of previous tick detection. B. burgdorferi was detected at nine sites, eight of which tested positive for the first time during this study and five of which had B. burgdorferi detected concurrently with initial tick detection. Tick and pathogen hotspots were identified in eastern Ontario in 2017 and 2018, respectively. These findings provide additional evidence on the range expansion and population establishment of I. scapularis in Ontario and help generate hypotheses on the invasion of B. burgdorferi in Ontario. Ongoing public health surveillance is critical to monitor changes in I. scapularis and its pathogens in Ontario.

Suppressive effects of Ixodes persulcatus sialostatin L2 against Borrelia miyamotoi-stimulated immunity.

Borrelia miyamotoi infection is an emerging tick-borne disease that causes hard tick-borne relapsing fever. B. miyamotoi is transmitted through the bite of ticks, including Ixodes persulcatus. Although accumulating evidence suggests that tick salivary proteins enhance the infectivity of other tick-borne pathogens, the association of B. miyamotoi with tick-derived proteins remains unknown. In this study, the effect of I. persulcatus sialostatin L2 (Ip-sL2), a tick-derived cystatin, on specific immunity to B. miyamotoi was preliminarily investigated in vitro. Mice were immunized with heat-killed B. miyamotoi and in vitro analyses of the splenocytes of the immunized mice indicated that the expression levels of the activation markers of CD11c+ and CD3+ cells were significantly upregulated by B. miyamotoi stimulation. Spleen cells from B. miyamotoi-immunized mice were used to determine whether Ip-sL2 regulates murine immune responses against B. miyamotoi. Treatment with Ip-sL2 in vitro inhibited the activation of CD11c+ and CD3+ cells as well as inflammatory cytokine production by cultured splenocytes. These findings show that Ip-sL2 has modulatory effects on murine immune responses to B. miyamotoi. Therefore, it is necessary to clarify in the future whether Ip-sL2 is involved in the enhanced infectivity of B. miyamotoi.

Tick infestation effects on haemoglobin levels of deer mice (Peromyscus maniculatus).

Deer mice (Peromyscus maniculatus) are hosts to ixodid ticks as well as the associated tick-borne pathogens they can spread. As the ranges of black-legged ticks (Ixodes scapularis) and American dog ticks (Dermacentor variabilis) expand northwards, naïve host populations of deer mice are likely to become infested by ticks and experience the physiological effects that ticks can have on them via blood-feeding. The prevalence of these haematophagous ticks can affect the haemoglobin levels of the mice they infest. Haemoglobin levels were compared and analysed in deer mice populations at three different sites with varying tick exposure. These results suggested that without confounding effects, the abundance of black-legged and American dog ticks on individual mice had a significant negative effect on the hosts' haemoglobin levels, but only in an area with high tick infestation. This was seen across the average haemoglobin levels between populations, where there was a significant difference between the source population with the longest established tick populations and the source population where neither black-legged nor American dog ticks were prevalent. As the ticks' ranges expand and they become more abundant, it is important to understand how their prevalence and intensity can alter host physiology, potentially affecting their own range expansion and the spread of the diseases they may carry.