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Introduction: The expanding geographical range of blacklegged ticks (BLTs), Ixodes scapularis, and its ability to transmit Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti, and Borrelia miyamotoi poses an emerging public health risk. Our study determined the geographic distribution and the minimum infection rate (MIR) of B. burgdorferi-, A. phagocytophilum-, Ba. microti-, and B. miyamotoi-infected BLTs in Manitoba submitted to the Public Health Agency of Canada's passive tick surveillance programme from 1995 to 2017., Methods: Regression models were used to test the association of the MIR by year for each pathogen. Ticks were tested using PCR for B. burgdorferi since 1995, A. phagocytophilum since 2006, and Ba. microti and B. miyamotoi since 2013. The global positioning system coordinates of infected and uninfected ticks submitted during the surveillance period were plotted on a map of Manitoba using ArcGIS Pro version 3.1.2 to detect changes in the geographic distribution of ticks over time., Results: The overall MIR for B. burgdorferi was 139.7 (95% confidence interval [CI]: 129.0-150.5) per 1000 BLTs; however, it varied over time. After remaining stable from 1995 to 2005, the MIR increased by 12.1 per 1000 BLTs per year from 2005 to 2017 (95% CI: 7.0%-17.2%, p-value <0.01). The geographic distribution of B. burgdorferi-infected BLTs was centred around Winnipeg, Manitoba, and spread outward from this locality. The MIRs of A. phagocytophilum, Ba. microti, and B. miyamotoi were 44.8 per 1000 BLTs (95% CI: 38.1-51.6), 10.8 (95% CI: 6.6-15.0), and 5.2 (95% CI: 2.3-8.1) per 1000 BLTs, respectively, and showed no significant change over time., Conclusion: Passive surveillance revealed the presence of A. phagocytophilum-, Ba. microti-, and B. miyamotoi-infected BLTs in southern Manitoba and revealed an increased risk of exposure to B. burgdorferi-infected BLTs due to the increasing geographic range and MIR., (© 2024 Wiley‐VCH GmbH. Published by John Wiley & Sons Ltd.)

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., (© 2022. The Author(s).)

Lyme disease (LD) risk is emerging rapidly in Canada due to range expansion of its tick vectors, accelerated by climate change. The risk of contracting LD varies geographically due to variability in ecological characteristics that determine the hazard (the densities of infected host-seeking ticks) and vulnerability of the human population determined by their knowledge and adoption of preventive behaviors. Risk maps are commonly used to support public health decision-making on Lyme disease, but the ability of the human public to adopt preventive behaviors is rarely taken into account in their development, which represents a critical gap. The objective of this work was to improve LD risk mapping using an integrated social-behavioral and ecological approach to: (i) compute enhanced integrated risk maps for prioritization of interventions and (ii) develop a spatially-explicit assessment tool to examine the relative contribution of different risk factors. The study was carried out in the Estrie region located in southern Québec. The blacklegged tick, Ixodes scapularis, infected with the agent of LD is widespread in Estrie and as a result, regional LD incidence is the highest in the province. LD knowledge and behaviors in the population were measured in a cross-sectional health survey conducted in 2018 reaching 10,790 respondents in Estrie. These data were used to create an index for the social-behavioral component of risk in 2018. Local Empirical Bayes estimator technique were used to better quantify the spatial variance in the levels of adoption of LD preventive activities. For the ecological risk analysis, a tick abundance model was developed by integrating data from ongoing long-term tick surveillance programs from 2007 up to 2018. Social-behavioral and ecological components of the risk measures were combined to create vulnerability index maps and, with the addition of human population densities, prioritization index maps. Map predictions were validated by testing the association of high-risk areas with the current spatial distribution of human cases of LD and reported tick exposure. Our results demonstrated that social-behavioral and ecological components of LD risk have markedly different distributions within Estrie. The occurrence of human LD cases or reported tick exposure in a municipality was positively associated with tick density and the prioritization risk index (p < 0.001). This research is a second step towards a more comprehensive integrated LD risk assessment approach, examining social-behavioral risk factors that interact with ecological risk factors to influence the management of emerging tick-borne diseases, an approach that could be applied more widely to vector-borne and zoonotic diseases., Competing Interests: Declaration of Competing Interest None., (Crown Copyright © 2022. Published by Elsevier GmbH. All rights reserved.)

ABSTRACT The geographic range and occurrence of tick species is dynamic. This has important public health implications due to important tick species that can transmit pathogens. This study presents a retrospective review of tick genera recovered from humans and submitted for identification in Alberta, Canada, over a 19-year period. The total number of ticks and proportion of genera were analyzed over time. Molecular testing for a number of pathogens associated with Ixodes scapularis and I. pacificus was conducted. A total of 2,358 ticks were submitted between 2000 and 2019, with 98.6% being acquired in Alberta. The number of ticks submitted increased significantly over time ( p  < 0.0001). Dermacentor ticks were the most abundant genus, followed by Ixodes and Amblyomma . There was a significant decrease in the proportion of Dermacentor ticks between 2013 and 2019 ( p  = 0.02), with a corresponding increase in the proportion of Ixodes ticks over the same time ( p  = 0.04). No statistically significant change in seasonality was identified. Borrelia burgdorferi was detected in 8/76 (10.5%; 95% CI 5.4-19.4%) of all I. scapularis and I. pacificus ticks submitted. This translated to a B. burgdorferi positivity of 0.35% (95% CI 0.15-0.68%) among all ticks received. Dermacentor species (especially D. andersoni ) remains the most common tick feeding on humans in Alberta. Small numbers of vector species (including I. scapularis/pacificus ) are encountered annually over widely separated geographic areas in the province. The risk of exposure to tick-borne pathogens (e.g. Lyme disease) in Alberta remains low.

The development of interventions that reduce Lyme disease incidence remains a challenge. Reservoir-targeted approaches aiming to reduce tick densities or tick infection prevalence with Borrelia burgdorferi have emerged as promising ways to reduce the density of infected ticks. Acaricides of the isoxazoline family offer high potential for reducing infestation of ticks on small mammals as they have high efficacy at killing feeding ticks for a long period. Fluralaner baits were recently demonstrated as effective, in the laboratory, at killing Ixodes scapularis larvae infesting Peromyscus mice, the main reservoir for B. burgdorferi in northeastern North America. Here, effectiveness of this approach for reducing the infestation of small mammals by immature stages of I. scapularis was tested in a natural environment. Two densities of fluralaner baits (2.1 baits/1,000 m2 and 4.4 baits/1,000 m2) were used during three years in forest plots. The number of I. scapularis larvae and nymphs per mouse from treated and control plots were compared. Fluralaner baiting reduced the number of larvae per mouse by 68% (CI95: 51-79%) at 2.1 baits/1,000 m2 and by 86% (CI95: 77-92%) at 4.4 baits/1,000 m2. The number of nymphs per mouse was reduced by 72% (CI95: 22-90%) at 4.4 baits/1,000 m2 but was not significantly reduced at 2.1 baits/1,000 m2. Reduction of Peromyscus mouse infestation by immature stages of I. scapularis supports the hypothesis that an approach targeting reservoirs of B. burgdorferi with isoxazolines has the potential to reduce tick-borne disease risk by decreasing the density of infected ticks in the environment., (© Crown copyright 2022.)

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.

Ixodes scapularis ticks are expanding their range in parts of northeastern North America, bringing with them pathogens of public health concern. While rodents like the white-footed mouse, Peromyscus leucopus, are considered the primary reservoir of many emerging tick-borne pathogens, the contribution of birds, as alternative hosts and reservoirs, to local transmission cycles has not yet been firmly established. From 2016 to 2018, we collected host-seeking ticks and examined rodent and bird hosts for ticks at 48 sites in a park where blacklegged ticks are established in Quebec, Canada, in order to characterize the distribution of pathogens in ticks and mammalian and avian hosts. We found nearly one third of captured birds (n = 849) and 70% of small mammals (n = 694) were infested with I. scapularis. Five bird and three mammal species transmitted Borrelia burgdorferi to feeding larvae (n larvae tested = 2257) and we estimated that about one fifth of the B. burgdorferi-infected questing nymphs in the park acquired their infection from birds, the remaining being attributable to mice. Ground-foraging bird species were more parasitized than other birds, and species that inhabited open habitat were more frequently infested and were more likely to transmit B. burgdorferi to larval ticks feeding upon them. Female birds were more likely to transmit infection than males, without age differentiation, whereas in mice, adult males were more likely to transmit infection than juveniles and females. We also detected Borrelia miyamotoi in larvae collected from birds, and Anaplasma phagocytophilum from a larva collected from a white-footed mouse. This study highlights the importance of characterising the reservoir potential of alternative reservoir hosts and to quantify their contribution to transmission dynamics in different species assemblages. This information is key to identifying the most effective host-targeted risk mitigation actions., Competing Interests: The authors have declared that no competing interests exist.

The tick vector of Lyme disease, Ixodes scapularis, is currently expanding its geographical distribution northward into southern Canada driving emergence of Lyme disease in the region. Despite large-scale studies that attributed different factors such as climate change and changes in land use to the geographical expansion of the tick, a comprehensive understanding of local patterns of tick abundance is still lacking in that region. Using a newly endemic periurban nature park located in Quebec (Canada) as a model, we explored intra-habitat patterns in tick distribution and their relationship with biotic and abiotic factors. We verified the hypotheses that (1) there is spatial heterogeneity in tick densities at the scale of the park and (2) these patterns can be explained by host availability, habitat characteristics and microclimatic conditions. During tick activity season in three consecutive years, tick, deer, rodent and bird abundance, as well as habitat characteristics and microclimatic conditions, were estimated at thirty-two sites. Patterns of tick distribution and abundance were investigated by spatial analysis. Generalised additive mixed models were constructed for each developmental stage of the tick and the relative importance of significant drivers on tick abundance were derived from final models. We found fine-scale spatial heterogeneity in densities of all tick stages across the park, with interannual variability in the location of hotspots. For all stages, the local density was related to the density of the previous stage in the previous season, in keeping with the tick's life cycle. Adult tick density was highest where drainage was moderate (neither waterlogged nor dry). Microclimatic conditions influenced the densities of immature ticks, through the effects of weather at the time of tick sampling (ambient temperature and relative humidity) and of the seasonal microclimate at the site level (degree-days and number of tick adverse moisture events). Seasonal phenology patterns were generally consistent with expected curves for the region, with exceptions in some years that may be attributable to founder events. This study highlights fine scale patterns of tick population dynamics thus providing fundamental knowledge in Lyme disease ecology and information applicable to the development of well-targeted prevention and control strategies for public natural areas affected by this growing problem in southern Canada., (Copyright © 2021. Published by Elsevier GmbH.)

Background: The universal nature of the human-companion animal relationship and their shared ticks and tick-borne pathogens offers an opportunity for improving public and veterinary health surveillance. With this in mind, we describe the spatiotemporal trends for blacklegged tick (Ixodes scapularis) submissions from humans and companion animals in Ontario, along with pathogen prevalence., Methods: We tested tick samples submitted through passive surveillance (2011-2017) from humans and companion animals for Borrelia burgdorferi, Borrelia miyamotoi, Anaplasma phagocytophilum and Babesia microti. We describe pathogen prevalence in ticks from humans and from companion animals and constructed univariable Poisson and negative binomial regression models to explore the spatiotemporal relationship between the rates of tick submissions by host type., Results: During the study, there were 17,230 blacklegged tick samples submitted from humans and 4375 from companion animals. Tick submission rates from companion animals were higher than expected in several public health units (PHUs) lacking established tick populations, potentially indicating newly emerging populations. Pathogen prevalence in ticks was higher in PHUs where established blacklegged tick populations exist. Borrelia burgdorferi prevalence was higher in ticks collected from humans (maximum likelihood estimate, MLE = 17.5%; 95% confidence interval, CI 16.97-18.09%) than from companion animals (9.9%, 95% CI 9.15-10.78%). There was no difference in pathogen prevalence in ticks by host type for the remaining pathogens, which were found in less than 1% of tested ticks. The most common co-infection B. burgdorferi + B. miyamotoi occurred in 0.11% of blacklegged ticks from humans and animals combined. Borrelia burgdorferi prevalence was higher in unengorged (21.9%, 95% CI 21.12-22.65%) than engorged ticks (10.0%, 95% CI 9.45-10.56%). There were no consistent and significant spatiotemporal relationships detected via regression models between the annual rates of submission of each host type., Conclusions: While B. burgdorferi has been present in blacklegged ticks in Ontario for several decades, other tick-borne pathogens are also present at low prevalence. Blacklegged tick and pathogen surveillance data can be used to monitor risk in human and companion animal populations, and efforts are under consideration to unite surveillance efforts for the different target populations.

Using next-generation sequencing DNA barcoding, we aimed to determine: 1) if the larval bloodmeal can be detected in Ixodes scapularis nymphs and 2) the post-moult temporal window for detection of the larval bloodmeal. Subsets of 30 nymphs fed on a domestic rabbit (Oryctolagus cuniculus Linnaeus, Lagomorphia: Leporidae) as larvae were reared and frozen at 11 time points post-moult, up to 150 d. Vertebrate DNA was amplified using novel universal (UP) and species-specific primers (SSP) and sequenced for comparison against cytochrome c oxidase subunit I barcodes to infer host identification. Detectable bloodmeals decreased as time since moult increased for both assays. For the SSP assay, detection of bloodmeals decreased from 96.7% (n = 29/30) in day 0 nymphs to 3.3% (n = 1/30) and 6.7% (n = 2/30) at 4- and 5-mo post-moult, respectively. A shorter temporal detection period was achieved with the UP assay, declining from 16.7% (n = 5/30) in day 0 nymphs to 0/30 in 3-d-old nymphs. Bloodmeal detection was nonexistent for the remaining cohorts, with the exception of 1/30 nymphs at 2-mo post-moult. Host detection was significantly more likely using the SSP assay compared to the UP assay in the first three time cohorts (day 0: χ 2 = 39.1, P < 0.005; day 2: χ 2 = 19.2, P < 0.005; day 3: χ 2 = 23.3, P < 0.005). Regardless of the primer set used, the next-generation sequencing DNA barcoding assay was able to detect host DNA from a larval bloodmeal in the nymphal life stage; however, a short window with a high proportion of detection post-moult was achieved., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Objective: Our objective was to apply next-generation sequence-based DNA barcoding to identify the remnant larval bloodmeals in wild-caught host-seeking (unengorged) Ixodes scapularis nymphs (n = 216). To infer host species identification, vertebrate DNA was amplified using universal primers for cytochrome c oxidase subunit I (COI) and sequenced using next-generation sequencing (NGS) for comparison against known barcode references., Results: Bloodmeal identification was unsuccessful in most samples (99% of 216 specimens) demonstrating a very low detection rate of this assay. Sequences that surpassed quality thresholds were obtained for 41.7% of nymphs (n = 90) and of those, confident species identification was obtained for 15.6% of nymphs (n = 14). Wild host identifications were only obtained from 2 specimens, where DNA from the eastern grey squirrel (Sciurus carolinensis) was identified. Human and bovine DNA was identified in remaining nymphs and considered to be contaminants. Further optimization of the technique is required to improve detection of remnant bloodmeals in host-seeking nymphs.

Canadians face an emerging threat of Lyme disease due to the northward expansion of the tick vector, Ixodes scapularis. We evaluated the degree of I. scapularis population establishment and Borrelia burgdorferi occurrence in the city of Ottawa, Ontario, Canada from 2017-2019 using active surveillance at 28 sites. We used a field indicator tool developed by Clow et al. to determine the risk of I. scapularis establishment for each tick cohort at each site using the results of drag sampling. Based on results obtained with the field indicator tool, we assigned each site an ecological classification describing the pattern of tick colonization over two successive cohorts (cohort 1 was comprised of ticks collected in fall 2017 and spring 2018, and cohort 2 was collected in fall 2018 and spring 2019). Total annual site-specific I. scapularis density ranged from 0 to 16.3 ticks per person-hour. Sites with the highest density were located within the Greenbelt zone, in the suburban/rural areas in the western portion of the city of Ottawa, and along the Ottawa River; the lowest densities occurred at sites in the suburban/urban core. B. burgdorferi infection rates exhibited a similar spatial distribution pattern. Of the 23 sites for which data for two tick cohorts were available, 11 sites were classified as "high-stable", 4 were classified as "emerging", 2 were classified as "low-stable", and 6 were classified as "non-zero". B. burgdorferi-infected ticks were found at all high-stable sites, and at one emerging site. These findings suggest that high-stable sites pose a risk of Lyme disease exposure to the community as they have reproducing tick populations with consistent levels of B. burgdorferi infection. Continued surveillance for I. scapularis, B. burgdorferi, and range expansion of other tick species and emerging tick-borne pathogens is important to identify areas posing a high risk for human exposure to tick-borne pathogens in the face of ongoing climate change and urban expansion., Competing Interests: The authors have declared that no competing interests exist.

Human granulocytic anaplasmosis is an obligate intra-granulocytic parasite that is transmitted by Ixodes scapularis and Ixodes pacificus in North America. We report on the second laboratory-confirmed case of Anaplasma phagocytophilum acquired within the province of Alberta, Canada. A 67-year-old woman from the Edmonton health zone developed nonspecific systemic symptoms including fatigue, night sweats, myalgia, headaches, and fever 6 days after noticing a tick on her left upper arm in May of 2017 (day 0). On day 13, she was found to have thrombocytopenia. Her symptoms progressed until day 16 when she was treated empirically with doxycycline, at which time she slowly improved over the subsequent 2 months. The tick was later identified as a partially engorged female blacklegged tick, I. scapularis , and it was positive for A. phagocytophilum DNA by PCR. Anaplasma serology performed retrospectively on blood samples collected on days 13, 31, and 52 showed a greater than 4-fold increase in A. phagocytophilum (IgG titers from less than 1:64 on day 13 to 1:2048 on days 31 and 52), consistent with an acute infection. Although populations of blacklegged ticks are not yet established in Alberta, suspicion should remain for tick-borne diseases because infected ticks are introduced into the province by migrating birds. This case report highlights the need to remind physicians and other public health professionals that rare, non-endemic tick-borne diseases can occasionally occur in low-risk jurisdictions.

Background: Lyme disease (LD) is an increasing public health threat in temperate zones of the northern hemisphere, yet relatively few methods exist for reducing LD risk in endemic areas. Disrupting the LD transmission cycle in nature is a promising avenue for risk reduction. This experimental study evaluated the efficacy of fluralaner, a recent oral acaricide with a long duration of effect in dogs, for killing Ixodes scapularis ticks in Peromyscus maniculatus mice, a known wildlife reservoir for Borrelia burgdorferi in nature., Methods: We assigned 87 mice to 3 fluralaner treatment groups (50 mg/kg, 12.5 mg/kg and untreated control) administered as a single oral treatment. Mice were then infested with 20 Ixodes scapularis larvae at 2, 28 and 45 days post-treatment and we measured efficacy as the proportion of infesting larvae that died within 48 h. At each infestation, blood from 3 mice in each treatment group was tested to obtain fluralaner plasma concentrations (C p )., Results: Treatment with 50 mg/kg and 12.5 mg/kg fluralaner killed 97% and 94% of infesting larvae 2 days post-treatment, but no significant effect of treatment on feeding larvae was observed 28 and 45 days post-treatment. Mouse C p did not differ significantly between the two tested doses. Mean C p decreased from 13,000 ng/ml in the 50 mg/kg group and 4000 ng/ml in the 12.5 mg/kg group at Day 2 to < 100 ng/ml in both groups at Day 45., Conclusions: We provide the first evidence that fluralaner is effective for killing immature ticks in Peromyscus mice, a first step in evaluating its potential for treating wild rodents as a public health intervention to reduce LD risk in endemic areas.

Passive and active surveillance for the blacklegged tick, Ixodes scapularis Say, in the Canadian province of Saskatchewan was conducted over a 9-yr period (2009-2017). More than 26,000 ixodid ticks, representing 10 species, were submitted through passive surveillance. Most (97%) of these were the American dog tick, Dermacentor variabilis (Say). Of the 65 I. scapularis adults submitted, 75% were collected from dogs. Infection rates of Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti in I. scapularis were 12%, 8%, and 0%, respectively. Although the I. scapularis submitted by passive surveillance were collected from five of seven ecoregions in central and southern Saskatchewan, they were most frequent in the Moist Mixed Grassland and Aspen Parklands. In contrast, no I. scapularis were collected from the extensive field sampling conducted at multiple sites in different ecoregions across the province. Hence, there is no evidence of I. scapularis having established a breeding population in Saskatchewan. Nonetheless, continued surveillance for blacklegged ticks is warranted given their important role as a vector of medically and veterinary important pathogens, and because they have recently become established across much of the southern portions of the neighboring province of Manitoba., (© Her Majesty the Queen in Right of Canada, as represented by the Minister of Agriculture and Agri-Food Canada, 2019.)

Human granulocytic anaplasmosis (HGA), caused by the bacteria Anaplasma phagocytophilum , is transmitted to humans by blacklegged ticks ( Ixodes scapularis ) in eastern North America. To assess the emergence of A . phagocytophilum in Ontario, we analyzed patient serological and clinical data in combination with pathogen detection in blacklegged ticks from 2011 to 2017. Our sample population included all patients who had Anaplasma serological testing ordered by their physicians ( n = 851). Eighty-three patients (10.8%) were A . phagocytophilum seropositive (IgG titers ≥ 1:64) and 686 (89.2%) were seronegative (IgG titers < 1:64). Applying published surveillance case definitions, we classified zero as confirmed, five as probable, and 78 as suspected cases. The percentage of seropositive patients remained generally stable at 13.6%. Seropositive patients were most often adult females, 40-59 years of age, and reported nonspecific signs and symptoms, such as fatigue, headache, and fever. Higher seropositivity rates (≥ 1.5 patients per 100,000 population) occurred in eastern and northwestern Ontario. The percentage of A . phagocytophilum -positive blacklegged ticks, through passive and active surveillance, was 0.4 and 1.1%, respectively, and increased over time. Serological and entomological indicators of A . phagocytophilum activity increased in areas of the province with established blacklegged tick populations. The risk of HGA is presently low in Ontario; however, further research is required to document the epidemiology of HGA in the province. To minimize the impact of HGA emergence in Ontario, increased awareness and education of the public and health-care providers is recommended, with consideration to making HGA a reportable infection in Ontario.

The occurrence of Borrelia garinii in seabird ticks, Ixodes uriae, associated with different species of colonial seabirds has been studied since the early 1990s. Research on the population structure of this bacterium in ticks from seabird colonies in the northeastern Atlantic Ocean has revealed admixture between marine and terrestrial tick populations. We studied B. garinii genetic diversity and population structure in I. uriae collected from seabird colonies in the northwestern Atlantic Ocean, in Newfoundland and Labrador, Canada. We applied a multi-locus sequence typing (MLST) scheme to B. garinii found in ticks from four species of seabirds. The B. garinii strains found in this seabird colony ecosystem were diverse. Some were very similar to strains from Asia and Europe, including some obtained from human clinical samples, while others formed a divergent group specific to this region of the Atlantic Ocean. Our findings highlight the genetic complexity of B. garinii circulating in seabird ticks and their avian hosts but also demonstrate surprisingly close connections between B. garinii in this ecosystem and terrestrial sources in Eurasia. Genetic similarities among B. garinii from seabird ticks and humans indicate the possibility that B. garinii circulating within seabird tick-avian host transmission cycles could directly, or indirectly via connectivity with terrestrial transmission cycles, have consequences for human health., (Crown Copyright © 2019. Published by Elsevier GmbH. All rights reserved.)

We thank Mr [...].

In eastern North America, the blacklegged tick, Ixodes scapularis, is the vector for Borrelia burgdorferi sensu stricto and Anaplasma phagocytophilum, the causal agents for human and canine Lyme disease and anaplasmosis, respectively. The extensive range expansion of I. scapularis in Ontario is a growing veterinary and public health concern. However, there is limited information on the risk factors associated with I. scapularis carriage on dogs. Within an emerging area for Lyme disease risk in southeastern Ontario, we identified the tick species carried by dogs; determined the prevalence of B. burgdorferi and A. phagocytophilum in I. scapularis; and examined associations between pet demographics, travel history, and geographical location and the odds of: a dog carrying I. scapularis relative to other tick species (i.e., case-case design), and a removed I. scapularis being infected with B. burgdorferi. Seven species of ticks were collected from 543 companion dogs at 20 participating veterinary hospitals from April to December 2015. Ixodes scapularis were detected on 85.6% of parasitized dogs, and 7.5% of these dogs were carrying at least one B. burgdorferi-positive tick. Based on a multivariable logistic regression model, the odds of I. scapularis infestation relative to other tick species was significantly higher in fall and spring compared to summer, with closer proximity to Lake Ontario, in female compared to male dogs, in dogs weighing over 30 kg compared to lighter dogs, and in dogs that had not visited a farm in the 7 days prior to tick removal. Based on univariable exact logistic regression models, the odds of B. burgdorferi-positive I. scapularis carriage relative to B. burgdorferi-negative I. scapularis were significantly higher for dogs that traveled in the 14 days prior to tick removal, for those dogs under 1 year of age, and for those weighing less than 10 kg. This case-case study provides information for veterinarians and public health practitioners to help protect dogs and their owners from Lyme disease in southeastern Ontario., (Copyright © 2018 Elsevier GmbH. All rights reserved.)

Competing Interests: Competing interests: None declared.

The range of Ixodes scapularis is expanding in Ontario, increasing the risk of Lyme disease. As an effective public health response requires accurate information on disease distribution and areas of risk, this study aims to establish the geographic distribution of I. scapularis and its associated pathogen, B. burgdorferi , in northwestern Ontario. We assessed five years of active and passive tick surveillance data in northwestern Ontario. Between 2013 and 2017, 251 I. scapularis were submitted through passive surveillance. The submission rate increased over time, and the proportion infected with B. burgdorferi was 13.5%. Active tick surveillance from 2014 to 2016 found few I. scapularis specimens. In 2017, 102 I. scapularis were found in 10 locations around the city of Kenora; 60% were infected with B. burgdorferi , eight tested positive for A. phagocytophilum , and one for POWV. I. scapularis ticks were found in 14 locations within the Northwestern Health Unit area, with seven locations containing B. burgdorferi -positive ticks. We found abundant I. scapularis populations in the southern portion of northwestern Ontario and northward expansion is expected. It is recommended that I. scapularis populations continue to be monitored and mitigation strategies should be established for rural northern communities.

In North America, different strains of the Lyme disease-causing bacterium Borrelia burgdorferi sensu stricto cluster into phylogenetic groups that are associated with different levels of pathogenicity and, for some, specific rodent reservoir hosts. Here we explore whether landscape connectivity, by impacting host dispersal, influences B. burgdorferi s.s. spread patterns. This question is central to modelling spatial patterns of the spread of Lyme disease risk in the zone of northward range-expansion of B. burgdorferi s.s. in southeastern Canada where the study was conducted. We used multi-locus sequence typing (MLST) to characterise B. burgdorferi s.s. in positive ticks collected at 13 sites in southern Quebec, Canada during the early stages of B. burgdorferi s.s. invasion. We used mixed effects logistic regression to investigate whether landscape connectivity (probability of connectivity; PC) affected the probability that samples collected at different sites were of the same strain (MLST sequence type: ST). PC was calculated from a habitat map based on high spatial resolution (15 m) Landsat 8 imagery to identify woodland habitat that are preferred by rodent hosts of B. burgdorferi s.s. There was a significant positive association between the likelihood that two samples were of the same ST and PC, when PC values were grouped into three categories of low, medium and high. When analysing data for individual STs, samples at different sites were significantly more likely to be the same when PC was higher for the rodent-associated ST1. These findings support the hypothesis that dispersion trajectories of B. burgdorferi s.s. in general, and some rodent-associated strains in particular, are at least partly determined by landscape connectivity. This may suggest that dispersion of B. burgdorferi s.s. is more common by terrestrial mammal hosts (which would likely disperse according to landscape connectivity) than by birds, the dispersal of which is likely less constrained by landscape. This study suggests that accounting for landscape connectivity may improve model-based predictions of spatial spread patterns of B. burgdorferi s.s. The findings are consistent with possible past dispersal patterns of B. burgdorferi s.s. as determined by phylogeographic studies., (Crown Copyright © 2018. Published by Elsevier GmbH. All rights reserved.)

Human granulocytic anaplasmosis (HGA) is an emerging disease in Canada because of range expansion by the arthropod vector, Ixodes scapularis. These ticks carry the Ap-ha variant of Anaplasma phagocytophilum (Ap-ha), which has been implicated in causing HGA, and the Ap-variant 1, which is not associated with human infection. We report the detection of 13 genotypes of the ankyrin (ankA) gene among 76 infected blacklegged ticks. Haplotype network and phylogenetic analyses revealed that the ankA genotypes corresponding to the Ap-ha variant did not form a monophyletic assemblage. They formed two distinct clades (Clades I and III), one of which was genetically more similar in nucleotide and amino acid sequences to genotypes of Ap-variant 1 that comprised Clade II. Additional work is needed to explore the evolutionary history of A. phagocytophilum in North America, and to determine if there are differences in pathogenicity or clinical symptoms associated with the two divergent groups of the Ap-ha variant given the significant differences in ankA amino acid sequence., (Crown Copyright © 2018. Published by Elsevier GmbH. All rights reserved.)

Lyme disease is an emerging public health threat in Canada. In this context, rapid detection of new risk areas is essential for timely application of prevention and control measures. In Canada, information on Lyme disease risk is collected through three surveillance activities: active tick surveillance, passive tick surveillance, and reported human cases. However, each method has shortcomings that limit its ability to rapidly and reliably identify new risk areas. We investigated the relationships between risk signals provided by human cases, passive and active tick surveillance to assess the performance of tick surveillance for early detection of emerging risk areas. We used regression models to investigate the relationships between the reported human cases, Ixodes scapularis (Say; Acari: Ixodidae) ticks collected on humans through passive surveillance and the density of nymphs collected by active surveillance from 2009 to 2014 in the province of Quebec. We then developed new risk indicators and validated their ability to discriminate risk levels used by provincial public health authorities. While there was a significant positive relationship between the risk signals provided all three surveillance methods, the strongest association was between passive tick surveillance and reported human cases. Passive tick submissions were a reasonable indicator of the abundance of ticks in the environment (sensitivity and specificity [Se and Sp] < 0.70), but were a much better indicator of municipalities with more than three human cases reported over 5 yr (Se = 0.88; Sp = 0.90). These results suggest that passive tick surveillance provides a timely and reliable signal of emerging risk areas for Lyme disease in Canada.

Since its detection in Canada in the early 1990s, Ixodes scapularis , the primary tick vector of Lyme disease in eastern North America, has continued to expand northward. Estimates of the tick's broad-scale distribution are useful for tracking the extent of the Lyme disease risk zone; however, tick distribution may vary widely within this zone. Here, we investigated I. scapularis nymph distribution at three spatial scales across the Lyme disease emergence zone in southern Quebec, Canada. We collected ticks and compared the nymph densities among different woodlands and different plots and transects within the same woodland. Hot spot analysis highlighted significant nymph clustering at each spatial scale. In regression models, nymph abundance was associated with litter depth, humidity, and elevation, which contribute to a suitable habitat for ticks, but also with the distance from the trail and the type of trail, which could be linked to host distribution and human disturbance. Accounting for this heterogeneous nymph distribution at a fine spatial scale could help improve Lyme disease management strategies but also help people to understand the risk variation around them and to adopt appropriate behaviors, such as staying on the trail in infested parks to limit their exposure to the vector and associated pathogens., Competing Interests: The authors declare no conflict of interest.

The emergence of the vector Ixodes scapularis in Ontario, Canada poses a significant public health risk. Both passive and active surveillance approaches have been employed by public health professionals (i.e., government employees) to monitor for the range expansion of this tick. Field surveillance using drag sampling for questing ticks is a recognized and effective method to identify reproducing tick populations. The degree of effort (i.e., number of visits per site) can enhance the sensitivity and specificity of surveillance, but increased effort conflicts with the cost to public health for field surveillance. Here we developed an indicator to determine the likelihood of I. scapularis establishment based on field sampling results. Field data from two established populations of I. scapularis in Ontario were incorporated with previous analyses of surveillance data to create the indicator, which is in the form of a scoring system. The life stage(s) collected, overall abundance and past surveillance findings from a site are all considered and a level is assigned for the likelihood of I. scapularis establishment based on current field sampling results. The likelihood levels are non-zero (i.e., no I. scapularis detected, but risk still present due to adventitious ticks), low, medium or high, and recommendations for future surveillance and public health measures are provided. The indicator was validated against field sampling results from five other established sites in the province and correctly categorized all five areas as high likelihood of establishment. The indicator was also applied to field sampling results from 36 sites of unknown status that were visited twice during the period of 2014-2016. There was substantial agreement of levels between measurements, as calculated using a weighted kappa. The indicator can assist public health professionals with the interpretation of field sampling results and direct their efforts for ongoing surveillance and public health interventions for I. scapularis-borne diseases, including Lyme disease.

The invasion of the blacklegged tick, Ixodes scapularis into Ontario, Canada poses a significant risk to public health because it is a vector for numerous pathogens, including Borrelia burgdorferi sensu stricto, the causative agent of Lyme disease. Baseline field sampling in 2014 and 2015 detected I. scapularis and B. burgdorferi at sites across southern, eastern and central Ontario, including a hot spot in eastern Ontario. A "speed of spread" model for I. scapularis developed by Leighton and colleagues (2012) estimated that the tick's range was expanding northward at 46 km/year. In 2016, we revisited a subset of sites sampled in 2014 and 2015 to understand the changing nature of risk, and assess whether the rate of tick invasion is consistent with the speed of spread estimate. Ticks were collected via tick dragging at 17 out of 36 sites, 5 of which were new sites for I. scapularis. Samples were positive for B. burgdorferi at 8 sites. No other I. scapularis-borne pathogens were detected. Centrographic statistics revealed an increase in the dispersion of I. scapularis positive sites in eastern Ontario. Field data for each site were then compared to the model's predicted year of establishment for each census subdivision. Our findings illustrate that the range expansion of I. scapularis and the emergence of B. burgdorferi is ongoing, and provide short timescale evidence of the processes associated with I. scapularis spread. The range front appears to be moving at a rate of ~46 km/year, with colonization of the tick behind this range front occurring at a slower and heterogeneous rate. Assessment of site-level ecological factors did not provide any insight into the underlying processes that may be influencing the colonization of I. scapularis in specific areas. Ongoing field sampling is needed to monitor this dynamic process. This study highlights the current geographic risk associated with Lyme disease, which can be used to target public health interventions to the areas of greatest risk.

Lyme disease is emerging in Canada due to geographic range expansion of the tick vector Ixodes scapularis Say. Recent areas of emergence include parts of the southeastern Canadian Prairie region. We developed a map of potential risk areas for future I. scapularis establishment in the Canadian Prairie Provinces. Six I. scapularis risk algorithms were developed using different formulations of three indices for environmental suitability: temperature using annual cumulative degree-days > 0 °C (DD > 0 °C; obtained from Moderate Resolution Imaging Spectroradiometer satellite data as an index of conditions that allow I. scapularis to complete its life cycle), habitat as a combined geolayer of forest cover and agricultural land use, and rainfall. The relative performance of these risk algorithms was assessed using receiver-operating characteristic (ROC) area under the curve (AUC) analysis with data on presence-absence of I. scapularis obtained from recent field surveillance in the Prairie Provinces accumulated from a number of sources. The ROC AUC values for the risk algorithms were significantly different (P < 0.01). The algorithm with six categories of DD > 0 °C, habitat as a simple dichotomous variable of presence or absence of forest, and normalized rainfall had the highest AUC of 0.74, representing "fair to good" performance of the risk algorithm. This algorithm had good (>80%) sensitivity in predicting positive I. scapularis surveillance sites, but low (50%) specificity as expected in this region where not all environmentally suitable habitats are expected to be occupied. Further prospective studies are needed to validate and perhaps improve the risk algorithm., (© Crown copyright 2017.)

Widespread access to the internet is offering new possibilities for data collection in surveillance. We explore, in this study, the possibility of using an electronic tool to monitor occurrence of the tick vector of Lyme disease, Ixodes scapularis. The study aimed to compare the capacity for ticks to be identified in web-based submissions of digital images/photographs, to the traditional specimen-based identification method used by the provincial public health laboratory in Quebec, Canada. Forty-one veterinary clinics participated in the study by submitting digital images of ticks collected from pets via a website for image-based identification by an entomologist. The tick specimens were then sent to the provincial public health laboratory to be identified by the 'gold standard' method using a microscope. Of the images submitted online, 74·3% (284/382) were considered of high-enough quality to allow identification. The laboratory identified 382 tick specimens from seven different species, with I. scapularis representing 76% of the total submissions. Of the 284 ticks suitable for image-based species identification, 276 (97·2%) were correctly identified (Kappa statistic of 0·92, Z = 15·46, P < 0·001). This study demonstrates that image-based tick identification may be an accurate and useful method of detecting ticks for surveillance when images are of suitable quality.

In northeastern North America, the blacklegged tick, Ixodes scapularis, is the vector of numerous tick-borne pathogens, including the agent of Lyme disease, Borrelia burgdorferi sensu stricto. Since 1990, there has been a rapid spread of I. scapularis northward into the province of Ontario, Canada. Climate change has been implicated as one of the driving factors for the spread of this vector. Other ecological factors also influence survival of I. scapularis populations and may facilitate invasion. The objective of this study was to identify local abiotic and biotic factors of significance for the invasion of I. scapularis in Ontario. The presence of ticks was determined by drag sampling at 154 sites in southern, eastern and central Ontario from May to October in 2014 and 2015. At each site, data on site aspect, forest cover, understory density and composition, soil moisture and composition, and the depth of litter layer were collected. Cumulative degree days above zero °C, total precipitation and elevation were attributed to each site using a geographic information system. A mixed multi-variable logistic regression model was created to assess the impact of the ecological factors on the presence of I. scapularis. In total, I. scapularis was found at 29 sites (18.8%) across the study area. The density of the understory, the presence of shrubs and the interaction of these two ecological factors were statistically significant, as well as longitude and cumulative degree days above zero. Our findings illustrate that local ecological factors are of importance for the invasion of I. scapularis into Ontario, and may be used to enhance local public health interventions and current predictive models and risk maps for I. scapularis., (Crown Copyright © 2017. Published by Elsevier GmbH. All rights reserved.)

Ixodes scapularis, the main vector of Borrelia burgdorferi, the spirochetal agent of Lyme disease, is expanding its range in southern Canada and bringing risk to the public from Lyme disease. The aims of this study were to (i) describe how risk of Lyme disease in Quebec, Canada, has changed from 2008 to 2014 by analysis of the number of tick submissions, the geographic scope of ticks submitted and the prevalence of B. burgdorferi in ticks removed from people and submitted through the Quebec passive tick surveillance program and (ii) explore whether exposure to ticks is influenced by age and sex. Ticks were collected from 2008 to 2014 in a passive surveillance program conducted by the Laboratoire de santé publique du Québec (LSPQ), and tested by PCR for B. burgdorferi at the National Microbiology Laboratory. The number of ticks submitted each year more than quadrupled during the study period (from 174 in 2008 to 962 in 2014), increases in the geographic range and geographic uniformity of submissions amongst municipalities were observed, and infection prevalence in the ticks (mostly adult females) submitted rose from 5.9% in 2008 to 18.1% in 2014. These data are consistent with outcomes from active surveillance for blacklegged ticks. More men (54.4%) than women (45.6%) were bitten by I. scapularis ticks and the frequency of tick submission was highest in children under 15 years of age and in the adults 50-70 years old. These findings demonstrate the utility of conducting passive tick surveillance using humans and provides information on risk groups (i.e., males, children under 15, adults older than 50, and those living in the more southern parts of the province) to which information on personal protection and tick-bite prevention should be most strongly targeted., (Crown Copyright © 2016. Published by Elsevier GmbH. All rights reserved.)

Lyme disease is emerging in Canada because of northward range expansion of the tick vector Ixodes scapularis. It is hypothesised that I. scapularis feeding on passerine birds migrating north in spring are important in founding new I. scapularis populations leading to northward range expansion. However, there are no studies on how far north I. scapularis may be carried, only inferences from passive tick surveillance. We used stable hydrogen isotope (δ(2)H) analysis of rectrices collected from northward migrating, I. scapularis-carrying, passerine birds captured in Canada to estimate how far north I. scapularis may be carried. Rectrices are usually grown close to breeding sites and their δ(2)H values reflect those in the environment, which vary strongly with latitude in North America. Passerines usually return to their breeding or natal sites so δ(2)H values of rectrices of northward migrating birds can identify the likely latitudinal bands of their intended destinations. In 2006 we analysed δ(2)H from rectrices of 73 I. scapularis-carrying birds captured at five migration monitoring stations, mainly from southern Ontario. Values of δ(2)H ranged from -33 to -124‰, suggesting 19/71 (26.7%) birds were destined for latitude band B (the most southerly part of Ontario), 40/71 (56.3%) birds were destined for band C (which extends from southern Ontario, Quebec and the Maritimes to southern James Bay) and 12/71 (16.9%) birds were destined for bands D and E (which extend from northern Ontario and Quebec into the southern Canadian Arctic). This indicates that many I. scapularis-carrying migratory birds in spring have destinations far north in Canada, including some farther north than the current region of climatic suitability for I. scapularis. These findings support the hypothesis that I. scapularis may continue to be spread north by spring migrating passerines. Some thrush species may be particularly implicated in far northward dispersion of I. scapularis., (Crown Copyright © 2015. Published by Elsevier GmbH. All rights reserved.)

Tick-borne diseases are a growing public health concern as their incidence and range have increased in recent decades. Lyme disease is an emerging infectious disease in Canada due to northward expansion of the geographic range of Ixodes scapularis, the principal tick vector for the Lyme disease agent Borrelia burgdorferi, into central and eastern Canada. In this study the geographical distributions of Ixodid ticks, including I. scapularis, and environmental factors associated with their occurrence were investigated in New Brunswick, Canada, where few I. scapularis populations have been found to date. Density of host-seeking ticks was evaluated by drag sampling of woodland habitats in a total of 159 sites. Ixodes scapularis ticks (n = 5) were found on four sites, Ixodes muris (n = 1) on one site and Haemaphysalis leporispalustris (n = 243) on 41 sites. One of four adult I. scapularis ticks collected was PCR-positive for B. burgdorferi. No environmental variables were significantly associated with the presence of I. scapularis although comparisons with surveillance data in neighbouring provinces (Québec and Nova Scotia) suggested that temperature conditions may be too cold for I. scapularis (< 2800 annual degree days above 0°C [DD > 0°C]) across much of New Brunswick. In contrast, the presence of H. leporispalustris, which is a competent vector of tularaemia, was significantly (P < 0.05) associated with specific ranges of mean DD > 0°C, mean annual precipitation, percentage of clay in site soil, elevation and season in a multivariable logistic regression model. With the exception of some localized areas, temperature conditions and deer density may be too low for the establishment of I. scapularis and Lyme disease risk areas in New Brunswick, while environmental conditions were suitable for H. leporispalustris at many sites. These findings indicate differing ecological niches for two tick species of public health significance.

The DNA sequences of the mitochondrial (mt) 12S rRNA and tRNA(Val) genes were characterized for 82 blacklegged ticks (Ixodes scapularis) that were genetically identical for Domains IV and V of the mt 16S rRNA gene. Thirty-one haplotypes, differed in sequence by 1-9 bp, were detected among the 82 ticks. Most nucleotide alterations in DNA sequence did not affect the stability of the secondary structures of the RNAs. The magnitude of the DNA sequence variation in the mt 12S rRNA and tRNA(Val) genes among blacklegged ticks suggests that this region of the mitochondrial genome has potential as a genetic marker for examining the population genetics and phylogeography of I. scapularis., (Copyright © 2015. Published by Elsevier Ltd.)

We developed PCR-based assays to distinguish a human pathogenic strain of Anaplasma phagocytophilum, Ap-ha, from Ap-variant 1, a strain not associated with human infection. The assays were validated on A. phagocytophilum-infected black-legged ticks (Ixodes scapularis) collected in Canada. The relative prevalence of these 2 strains in I. scapularis ticks differed among geographic regions.

Background: Ixodes scapularis is a vector of several human pathogens in the United States, and there is geographical variation in the relative number of persons infected with these pathogens. Geographically isolated populations of I. scapularis have established or are in the process of establishing in southern Canada. Knowledge of the genetic variation within and among these populations may provide insight into their geographical origins in the United States and the potential risk of exposure of Canadians to the different pathogens carried by I. scapularis., Methods: Part of the mitochondrial (mt) 16S ribosomal (r) RNA gene was amplified by PCR from 582 ticks collected from southern Canada, and Minnesota and Rhode Island in the United States. Sequence variation was examined in relation to the predicted secondary structure of the gene. Genetic diversity among populations was also determined., Results: DNA sequence analyses revealed 52 haplotypes. Most mutational alterations in DNA sequence occurred at unpaired sites or represented partial compensatory base pair changes that maintained the stability of the secondary structure. Significant genetic variation was detected within and among populations in different geographical regions. A greater proportion of the haplotypes of I. scapularis from the Canadian Prairie Provinces were found in the Midwest of the United States than in other regions, whereas more of the haplotypes of I. scapularis from the Canadian Central and Atlantic Provinces occurred in the Northeast of the United States. Nonetheless, 58% of I. scapularis were of a haplotype that occurs in the Midwest and Northeast of the United States; thus, their geographical origins could not be determined., Conclusions: There is considerable genetic variation in the mt 16S rRNA gene of I. scapularis. There is some evidence to support the hypothesis that some lineages of I. scapularis in the Atlantic and Central Provinces of Canada may be derived from colonizing individuals originating in the Northeast of the United States, whereas those in the Prairie Provinces may be derived from individuals originating in the Midwest of the United States. However, additional genetic markers are needed to test hypotheses concerning the geographical origins of I. scapularis in Canada.

Background: Blacklegged ticks, Ixodes scapularis are vectors of the tick-borne pathogens Borrelia burgdorferi, Anaplasma phagocytophilum and Babesia microti. Recently, the I. scapularis-borne bacterium Borrelia miyamotoi has been linked to human illness in North America. The range of this tick is expanding in Canada which may increase the potential for human exposure to these agents., Methods: In this study, 4938 I. scapularis ticks collected in 2012 were tested following a newly developed PCR-based testing protocol to determine the prevalence of infection with B. miyamotoi and other pathogens in I. scapularis in Canada., Results: Borrelia miyamotoi was detected in blacklegged ticks from all provinces except Newfoundland, although the infection prevalence was low (<1%). There was significant variation among provinces in the prevalence of infection of ticks with B. burgdorferi and A. phagocytophilum, but not with B. miyamotoi., Conclusions: Given the widespread distribution of B. miyamotoi, infection due to this agent should be considered in patients who have been exposed to blacklegged ticks in Canada.

Genotyping of Ixodes scapularis Say (Acari: Ixodidae) ticks could enhance understanding of the occurrence and genotypes of I. scapularis-borne pathogens. We investigated the utility of mitochondrial (mt) Cytochrome C Oxidase subunit I gene (cox1) sequences as a tool for understanding the population structure of I. scapularis collected in Canada, where we also investigated the geographic occurrence of different cox1 haplotypes. Sequences obtained from 414 ticks were one of 55 unique haplotypes, most of which grouped into one of six clades. Demographic analysis suggested that cox1 sequences have haplotype and nucleotide diversity comparable to other mt genes. All haplotypes were connected in a single minimum spanning network tree. Despite low fixation index values there were significant differences in the frequency of occurrence of haplotypes of different clades among four geographic regions: 1) Alberta to western Ontario, 2) eastern Ontario, 3) Quebec, and 4) Atlantic Provinces; suggesting that cox1 sequences could reveal population structure differences between I. scapularis in geographically separated populations of northeastern and midwestern North America. Spatial clusters of ticks of the same haplotype identified in regions of southern Quebec and southern Ontario where I. scapularis is invading were consistent with population bottlenecks associated with founder events. These findings suggest that cox1 sequences are useful for the study of I. scapularis population structure, are of sufficient diversity that spatial analyses of haplotypes can be used to identify where I. scapularis is emerging in southern Canada, and may be useful for exploring differences between northeastern and midwestern populations of I. scapularis.

Due to recent establishment of the blacklegged tick, Ixodes scapularis Say, in southeastern Canada, tick-borne zoonoses (Lyme disease, human granulocytotropic anaplasmosis, and babesiosis) are of growing concern for public health. Using white-tailed deer (Odocoileus virginianus) culled in southwestern Quebec during 2007-2008, we investigated whether hunter-killed deer could act as sentinels for early establishing tick populations and for tick-borne pathogens. Accounting for environmental characteristics of culling sites, and age and sex of deer, we investigated whether their tick infestation levels could identify locations of known tick populations detected in active surveillance, presumed tick populations detected by passive surveillance, or both. We also used spatial cluster analyses to identify spatial patterns of tick infestation and occurrence of tick-borne zoonoses infection in ticks collected from the deer. Adult ticks were found on 15% of the 583 deer examined. Adult male deer had the greatest number (approximately 90%) of adult ticks. Overall, 3, 15, and 0% of the ticks collected were polymerase chain reaction (PCR)-positive for Borrelia burgdorferi, Anaplasma phagocytophilum, and Babesia microti, respectively. Our statistical analyses suggest that sex and age of deer, temperature, precipitation, and an index of tick dispersion by migratory birds were significantly associated with tick infestation levels. Cluster analysis identified significant clusters of deer carrying ticks PCR-positive for A. phagocytophilum, and for deer carrying two or more I. scapularis. Our study suggests that hunter-killed deer may be effective as sentinels for emerging areas of tick-borne anaplasmosis. They may have limited use as sentinels for early emerging I. scapularis tick populations and emerging Lyme disease risk.

Lyme disease (LD) is emerging in Canada because of the northward expansion of the geographic range of the tick vector Ixodes scapularis (Say). Early detection of emerging areas of LD risk is critical to public health responses, but the methods to do so on a local scale are lacking. Passive tick surveillance has operated in Canada since 1990 but this method lacks specificity for identifying areas where tick populations are established because of dispersion of ticks from established LD risk areas by migratory birds. Using data from 70 field sites in Quebec visited previously, we developed a logistic regression model for estimating the risk of I. scapularis population establishment based on the number of ticks submitted in passive surveillance and a model-derived environmental suitability index. Sensitivity-specificity plots were used to select an optimal threshold value of the linear predictor from the model as the signal for tick population establishment. This value was used to produce an "Alert Map" identifying areas where the passive surveillance data suggested ticks were establishing in Quebec. Alert Map predictions were validated by field surveillance at 76 sites: the prevalence of established I. scapularis populations was significantly greater in areas predicted as high-risk by the Alert map (29 out of 48) than in areas predicted as moderate-risk (4 out of 30) (P < 0.001). This study suggests that Alert Maps created using this approach can provide a usefully rapid and accurate tool for early identification of emerging areas of LD risk at a geographic scale appropriate for local disease control and prevention activities.

Immature Ixodes scapularis infestation and Borrelia burgdorferi infection of wild small mammals were studied from June to October in 2007 and from May to October in 2008 at 71 study sites in a zone where I. scapularis populations and environmental Lyme disease risk are emerging in southwestern Quebec. Seasonal host-seeking activity of immature I. scapularis was similar to patterns reported previously in Canada and the USA: nymphal activity peaked in spring while larval activity peaked in late summer. Synchronous activity of nymphs with some larvae was observed in late spring, which could favour establishment of B. burgdorferi strains that cause short-lived infections in their hosts. White-footed mice (Peromyscus leucopus), deer mice (P. maniculatus), chipmunks (Tamias striatus), and red squirrels (Tamiasciurus hudsonicus) carried 92.0% of the larvae and 94.2% of the nymphs collected. Adult male white-footed mice carried significantly larger numbers of both larval and nymphal I. scapularis than other species and classes of small mammals (different demographic groups or physiological status: age, sex, sexual activity). We conclude that seasonality and host association were comparable to previous studies in North America, even in the context of a newly endemic pattern of low infection prevalence and low densities of host-seeking and feeding I. scapularis in southwestern Quebec. Our studies suggest that B. burgdorferi transmission cycles are focused on adult male mice (which carried 35% of all feeding ticks collected in the study), so control methods targeting this class of hosts may be particularly effective. However, our study also suggested that habitats containing a diverse host structure may dilute transmission cycles by partitioning of nymphal and larval ticks on different host species., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

Although Ixodes scapularis is the most important vector of Borrelia burgdorferi sensu stricto, the causative agent of Lyme disease in North America, little is known of the genetic diversity in this tick species within the recently established populations in Canada. In the present study, 153 I. scapularis adults collected from southern Canada were compared genetically using single-strand conformation polymorphism (SSCP) analyses in combination with DNA sequencing of the mitochondrial 16S rRNA gene. Nineteen haplotypes were detected, 8 of which have not been reported in the U.S.A. One 'new' haplotype was only detected at Lunenburg, Nova Scotia, and comprised 38% of the ticks examined for that population. The population in the southeast corner of Manitoba contained 3 'new' haplotypes. Although the most common haplotype (Is-1) was present in all 6 populations of I. scapularis in Canada, there were significant differences in the genetic structure among population. This suggests different geographical origins for the tick populations in Canada, which may be related to the transportation of larval and nymphal ticks by migratory passerines using different flyways. Determination of the origins of the endemic populations of I. scapularis in Canada, as well as those predicted to establish in the near future, has important implications with respect to understanding the risk of exposure to pathogenic bacteria infecting these ticks., (Copyright © 2011 Elsevier GmbH. All rights reserved.)

The genetic diversity of Borrelia burgdorferi sensu stricto, the agent of Lyme disease in North America, has consequences for the performance of serological diagnostic tests and disease severity. To investigate B. burgdorferi diversity in Canada, where Lyme disease is emerging, bacterial DNA in 309 infected adult Ixodes scapularis ticks collected in surveillance was characterized by multilocus sequence typing (MLST) and analysis of outer surface protein C gene (ospC) alleles. Six ticks carried Borrelia miyamotoi, and one tick carried the novel species Borrelia kurtenbachii. 142 ticks carried B. burgdorferi sequence types (STs) previously described from the United States. Fifty-eight ticks carried B. burgdorferi of 1 of 19 novel or undescribed STs, which were single-, double-, or triple-locus variants of STs first described in the United States. Clonal complexes with founder STs from the United States were identified. Seventeen ospC alleles were identified in 309 B. burgdorferi-infected ticks. Positive and negative associations in the occurrence of different alleles in the same tick supported a hypothesis of multiple-niche polymorphism for B. burgdorferi in North America. Geographic analysis of STs and ospC alleles were consistent with south-to-north dispersion of infected ticks from U.S. sources on migratory birds. These observations suggest that the genetic diversity of B. burgdorferi in eastern and central Canada corresponds to that in the United States, but there was evidence for founder events skewing the diversity in emerging tick populations. Further studies are needed to investigate the significance of these observations for the performance of diagnostic tests and clinical presentation of Lyme disease in Canada.

Background: Northward expansion of the tick Ixodes scapularis is driving Lyme disease (LD) emergence in Canada. Information on mechanisms involved is needed to enhance surveillance and identify where LD risk is emerging., Objectives: We used passive and active surveillance and phylogeographic analysis of Borrelia burgdorferi to investigate LD risk emergence in Quebec., Methods: In active surveillance, we collected ticks from the environment and from captured rodents. B. burgdorferi transmission was detected by serological analysis of rodents and by polymerase chain reaction assays of ticks. Spatiotemporal trends in passive surveillance data assisted interpretation of active surveillance. Multilocus sequence typing (MLST) of B. burgdorferi in ticks identified likely source locations of B. burgdorferi., Results: In active surveillance, we found I. scapularis at 55% of sites, and we were more likely to find them at sites with a warmer climate. B. burgdorferi was identified at 13 I. scapularis-positive sites, but infection prevalence in ticks and animal hosts was low. Low infection prevalence in ticks submitted in passive surveillance after 2004-from the tick-positive regions identified in active surveillance-coincided with an exponential increase in tick submissions during this time. MLST analysis suggested recent introduction of B. burgdorferi from the northeastern United States., Conclusions: These data are consistent with I. scapularis ticks dispersed from the United States by migratory birds, founding populations where the climate is warmest, and then establishment of B. burgdorferi from the United States several years after I. scapularis have established. These observations provide vital information for public health to minimize the impact of LD in Canada.

Background: Lyme disease is the commonest vector-borne zoonosis in the temperate world, and an emerging infectious disease in Canada due to expansion of the geographic range of the tick vector Ixodes scapularis. Studies suggest that climate change will accelerate Lyme disease emergence by enhancing climatic suitability for I. scapularis. Risk maps will help to meet the public health challenge of Lyme disease by allowing targeting of surveillance and intervention activities., Results: A risk map for possible Lyme endemicity was created using a simple risk algorithm for occurrence of I. scapularis populations. The algorithm was calculated for each census sub-division in central and eastern Canada from interpolated output of a temperature-driven simulation model of I. scapularis populations and an index of tick immigration. The latter was calculated from estimates of tick dispersion distances by migratory birds and recent knowledge of the current geographic range of endemic I. scapularis populations. The index of tick immigration closely predicted passive surveillance data on I. scapularis occurrence, and the risk algorithm was a significant predictor of the occurrence of I. scapularis populations in a prospective field study. Risk maps for I. scapularis occurrence in Canada under future projected climate (in the 2020s, 2050s and 2080s) were produced using temperature output from the Canadian Coupled Global Climate Model 2 with greenhouse gas emission scenario enforcing 'A2' of the Intergovernmental Panel on Climate Change., Conclusion: We have prepared risk maps for the occurrence of I. scapularis in eastern and central Canada under current and future projected climate. Validation of the risk maps provides some confidence that they provide a useful first step in predicting the occurrence of I. scapularis populations, and directing public health objectives in minimizing risk from Lyme disease. Further field studies are needed, however, to continue validation and refinement of the risk maps.

During the spring in 2005 and 2006, 39,095 northward-migrating land birds were captured at 12 bird observatories in eastern Canada to investigate the role of migratory birds in northward range expansion of Lyme borreliosis, human granulocytic anaplasmosis, and their tick vector, Ixodes scapularis. The prevalence of birds carrying I. scapularis ticks (mostly nymphs) was 0.35% (95% confidence interval [CI] = 0.30 to 0.42), but a nested study by experienced observers suggested a more realistic infestation prevalence of 2.2% (95% CI = 1.18 to 3.73). The mean infestation intensity was 1.66 per bird. Overall, 15.4% of I. scapularis nymphs (95% CI = 10.7 to 20.9) were PCR positive for Borrelia burgdorferi, but only 8% (95% CI = 3.8 to 15.1) were positive when excluding nymphs collected at Long Point, Ontario, where B. burgdorferi is endemic. A wide range of ospC and rrs-rrl intergenic spacer alleles of B. burgdorferi were identified in infected ticks, including those associated with disseminated Lyme disease and alleles that are rare in the northeastern United States. Overall, 1.4[corrected]% (95% CI = 0.3 [corrected] to 0.41) of I. scapularis nymphs were PCR positive for Anaplasma phagocytophilum. We estimate that migratory birds disperse 50 million to 175 million I. scapularis ticks across Canada each spring, implicating migratory birds as possibly significant in I. scapularis range expansion in Canada. However, infrequent larvae and the low infection prevalence in ticks carried by the birds raise questions as to how B. burgdorferi and A. phagocytophilum become endemic in any tick populations established by bird-transported ticks.

Fitness of tick-borne pathogens may be determined by the degree to which their infection dynamics in vertebrate hosts permits transmission cycles if infective and uninfected tick stages are active at different times of the year. To investigate this hypothesis we developed a simulation model that integrates the transmission pattern imposed by seasonally asynchronous nymphal and larval Ixodes scapularis ticks in northeastern North America, with a model of infection in white-footed mice (Peromyscus leucopus) reservoir hosts, using the bacteria Borrelia burgdorferi and Anaplasma phagocytophilum as examples. In simulations, survival of microparasites, their sensitivity to reduced rodent and tick abundance, and to 'dilution' by a reservoir-incompetent host depended on traits that allowed (i) highly efficient transmission from acutely-infected hosts, (ii) long-lived acute or 'carrier' host infections, and/or (iii) transmission amongst co-feeding ticks. Minimum values for transmission efficiency to ticks, and duration of host infectivity, necessary for microparasite persistence, were always higher when nymphal and larval ticks were seasonally asynchronous than when these instars were synchronous. Thus, traits influencing duration of host infectivity, transmission efficiency to ticks and co-feeding transmission are likely to be dominant determinants of fitness in I. scapularis-borne microparasites in northeastern North America due to abiotic forcings influencing I. scapularis seasonality.

Passive surveillance for the occurrence of the tick Ixodes scapularis Say (1821) and their infection with the Lyme borreliosis spirochaetes Borrelia burgdorferi s.l. has taken place in Canada since early 1990. Ticks have been submitted from members of the public, veterinarians, and medical practitioners to provincial, federal, and university laboratories for identification, and the data have been collated and B. burgdorferi detected at the National Microbiology Laboratory. The locations of collection of 2,319 submitted I. scapularis were mapped, and we investigated potential risk factors for I. scapularis occurrence (in Quebec as a case study) by using regression analysis and spatial statistics. Ticks were submitted from all provinces east of Alberta, most from areas where resident I. scapularis populations are unknown. Most were adult ticks and were collected in spring and autumn. In southern Québec, risk factors for tick occurrence were lower latitude and remote-sensed indices for land cover with woodland. B. burgdorferi infection, identified by conventional and molecular methods, was detected in 12.5% of 1,816 ticks, including 10.1% of the 256 ticks that were collected from humans and tested. Our study suggests that B. burgdorferi-infected I. scapularis can be found over a wide geographic range in Canada, although most may be adventitious ticks carried from endemic areas in the United States and Canada by migrating birds. The risk of Lyme borreliosis in Canada may therefore be mostly low but more geographically widespread than previously suspected.