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Lyme disease, caused by vector-borne Borrelia bacteria, can present with diverse multi-system symptoms that resemble other conditions. The objective of this study was to evaluate disease presentations and Borrelia seroreactivity in individuals experiencing a spectrum of chronic and complex illnesses. We recruited 157 participants from Eastern Canada who reported one or more diagnoses of Lyme disease, neurological, rheumatic, autoimmune, inflammatory, gastrointestinal, or cardiovascular illnesses, or were asymptomatic and presumed healthy. Intake categories were used to classify participants based on their perceived proximity to Lyme disease, distinguishing between those with a disclosed history of Borrelia infection, those with lookalike conditions (e.g. fibromyalgia syndrome), and those with unrelated ailments (e.g. intestinal polyps). Participants completed three questionnaires, the SF-36 v1, SIQR, and HMQ, to capture symptoms and functional burden, and provided blood serum for analysis at an accredited diagnostic lab. Two-tiered IgG and IgM serological assessments (whole cell ELISA and Western blot) were performed in a blinded fashion on all samples. The pattern of symptoms and functional burden were similarly profound in the presumptive Lyme and Lyme-like disease categories. Borrelia seroprevalence across the study cohort was 10% for each of IgG and IgM, and occurred within and beyond the Lyme disease intake category. Western blot positivity in the absence of reactive ELISA was also substantial. Fibromyalgia was the most common individual diagnostic tag disclosed by two-tier IgG-positive participants who did not report a history of Lyme disease. Within the IgG seropositive cohort, the presence of antibodies against the 31 kDa Outer Surface Protein A (OspA) was associated with significantly better health outcomes. Previously, this marker has been linked to treatment-refractory Lyme arthritis. Overall, our findings support prior observations of phenotypic overlap between Lyme and other diseases. Seropositivity associated with non-specific symptoms and functional impairment warrants further mechanistic investigation and therapeutic optimization., Competing Interests: The authors have declared that no competing interests exist., (Copyright: © 2023 Sanderson et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Borrelia burgdorferi and Borrelia miyamotoi are tick-vectored zoonotic pathogens maintained in wildlife species. Tick populations are establishing in new areas globally in response to climate change and other factors. New Brunswick is a Canadian maritime province at the advancing front of tick population establishment and has seen increasing numbers of ticks carrying B. burgdorferi, and more recently B. miyamotoi. Further, it is part of a region of Atlantic Canada with wildlife species composition differing from much of continental North America and little information exists as to the presence and frequency of infection of Borrelia spp. in wildlife in this region. We used a citizen science approach to collect a wide range of animals including migratory birds, medium-sized mammals, and small mammals. In total we tested 339 animals representing 20 species for the presence of B. burgdorferi and B. miyamotoi. We have developed new nested PCR primers and a protocol with excellent specificity for detecting both of these Borrelia species, both single and double infections, in tissues and organs of various wildlife species. The positive animals were primarily small non-migratory mammals, approximately twice as many were infected with B. burgdorferi than B. miyamotoi and one animal was found infected with both. In addition to established reservoir species, the jumping mouse (Napaeozapus insignis) was found frequently infected; this species had the highest infection prevalence for both B. burgdorferi and B. miyamotoi and has not previously been identified as an important carrier for either Borrelia species. Comprehensive testing of tissues found that all instances of B. burgdorferi infection were limited to one tissue within the host, whereas two of the five B. miyamotoi infections were diffuse and found in multiple systems. In the one coinfected specimen, two fetuses were also recovered and found infected with B. miyamotoi. This presumptive transplacental transmission suggests that vertical transmission in mammals is possible. This finding implies that B. miyamotoi could rapidly spread into wildlife populations, as well as having potential human health implications., Competing Interests: The authors have declared that no competing interests exist.

Lyme disease is a tick-borne disease that is emerging in Canada. The disease is caused by spirochetes of the Lyme borreliosis group, which is expanding as new species are discovered. In Canada, Lyme disease risk has so far been assessed primarily by detection of Borrelia burgdorferi sensu stricto. Of Ixodes scapularis ticks collected between 2014 and 2016 in New Brunswick, Canada, 7 were shown to be infected with Borrelia bissettii by nested PCR and sequencing of 5 B. bissettii genes. Since different Borrelia species are associated with different clinical manifestations and are not detected with the same diagnostic tests, the identification of a previously undocumented or underreported pathogenic Borrelia species has important implications for public and veterinary medicine.

Tick-borne pathogens are growing in importance for human and veterinary research worldwide. We developed, optimized, and validated a reliable quantitative PCR (qPCR; real-time PCR) assay to assess Borrelia burgdorferi infection by targeting two B. burgdorferi genes, ospA and flaB. When assessing previously tested tick samples, its performance surpassed the nested PCR in efficiency, sensitivity, and specificity. Since the detection of Borrelia is more difficult in mammalian samples, the qPCR assay was also assessed using wildlife tissues. For wildlife samples, the sensitivity and specificity of ospA primers, with the incorporation of a pre-amplification step, was equivalent or superior to the nested PCR. For human samples, no primer set was successful with human tissue without culture, but we detected Borrelia with ospA and flaB primers in 50% of the Lyme culture samples, corresponding to 60% of the participants with a Lyme disease diagnosis or suspicion. The specificity of amplification was confirmed by Sanger sequencing. The healthy participant culture samples were negative. This PCR-based direct detection assay performs well for the detection of Borrelia in different biological samples. Advancements in detection methods lead to a better surveillance of Borrelia in vectors and hosts, and, ultimately, enhance human and animal health. [ABSTRACT FROM AUTHOR]

Borrelia burgdorferi and Borrelia miyamotoi are tickborne zoonotic pathogens in Canada. Both bacteria are vectored by ticks, Ixodes scapularis in Atlantic Canada, but require wildlife reservoir species to maintain the bacteria for retransmission to future generations of ticks. Coyotes (Canis latrans) are opportunistic feeders, resulting in frequent contact with other animals and with ticks. Because coyotes are closely related to domestic dogs (Canis lupus familiaris), it is probable that coyote susceptibility to Borrelia infection is similar to that of dogs. We collected livers and kidneys of eastern coyotes from licensed harvesters in Nova Scotia, Canada, and tested them using nested PCR for the presence of B. burgdorferi, B. miyamotoi, and Dirofilaria immitis. Blood obtained from coyote livers was also tested serologically for antibodies to B. burgdorferi, Ehrlichia canis, Anaplasma phagocytophilum, and D. immitis. Borrelia burgdorferi and D. immitis were detected by both nested PCR and serology tests. Seroreactivity to A. phagocytophilum was also found. Borrelia miyamotoi and E. canis were not detected. Our results show that coyotes in Nova Scotia have been exposed to a number of vectorborne pathogens. [ABSTRACT FROM AUTHOR]

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Ticks are vectors of many diseases, including Lyme disease (Ld). Lyme disease is an emerging disease in Canada caused by infection with the Lyme borreliosis (Lb) members of the Borrelia genus of spirochaete bacteria, of which Borrelia burgdorferi is regionally the most prevalent. The primary tick vector in central and eastern Canada, Ixodes scapularis, is increasing in numbers and in the geographical extent of established populations. This study documents the distribution of ticks recovered by passive surveillance, and their B. burgdorferi infection prevalence, in three Canadian Maritime provinces from 2012–2020. These regions represent areas in which tick populations are widely established, establishing, and considered non-established. Using a community science approach by partnering with veterinarians and members of the public, we collected over 7000 ticks from the 3 provinces. The three species found most often on companion animals and humans were I. scapularis (76.9%), Ixodes cookei (10.4%) and Dermacentor variabilis (8.9%). The most common hosts were dogs (60.5%), cats (16.8%) and humans (17.6%). As is typical of passive surveillance tick collections, the majority of ticks recovered were adult females; for I. scapularis 90.2%, 5.3%, 3.9% and 0.6% of the total of 5630 ticks recovered for this species were adult females, adult males, nymphs and larvae, respectively. The majority of B. burgdorferi-infected ticks were I. scapularis, as expected. Borrelia infection prevalence in I scapularis was higher in Nova Scotia (20.9%), the province with the most endemic regions, than New Brunswick (14.1%) and Prince Edward Island (9.1%), provinces thought to have established and non-established tick populations, respectively. The province-wide Borrelia infection prevalence generally increased in these latter tow provinces over the course of the study. The host did not have a significant effect on B. burgdorferi infection prevalence; I. scapularis ticks from dogs, cats, humans was, 13.3% (n = 3622), 15.6% (n = 817), 17.9% (n = 730), respectively. No I. scapularis larvae were found infected (n = 33) but B. burgdorferi was detected in 14.8% of both adults (n = 5140) and nymphs (n = 215). The incidence of B. burgdorferi infection also did not differ by engorgement status 15.0% (n = 367), 15.1% (n = 3101) and 14.4% (n = 1958) of non-engorged, engorged and highly engorged ticks, respectively, were infected. In New Brunswick, at the advancing front of tick population establishment, the province-wide infection percentages generally increased over the nine-year study period and all health district regions showed increased tick recoveries and a trend of increased percentages of Borrelia-infected ticks over the course of the study. Within New Brunswick, tick recoveries but not Borrelia infection prevalence were significantly different from endemic and non-endemic regions, suggesting cryptic endemic regions existed prior to their designation as a risk area. Over the 9 years of the study, tick recoveries increased in New Brunswick, the primary study region, and I. scapularis recoveries spread northwards and along the coast, most but not all new sites of recoveries were predicted by climate-based models, indicating that ongoing tick surveillance is necessary to accurately detect all areas of risk. Comparison of tick recoveries and public health risk areas indicates a lag in identification of risk areas. Accurate and timely information on tick distribution and the incidence of Borrelia and other infections are essential for keeping the public informed of risk and to support disease prevention behaviors. [ABSTRACT FROM AUTHOR]