Draft whole-genome sequences of three Borrelia burgdorferi isolates from Western Canada.

Whole-genome sequences are presented for three Borrelia burgdorferi, a causative agent of Lyme disease in North America, isolated from Ixodes pacificus ticks collected in British Columbia, Canada. Shotgun DNA libraries were prepared with Illumina DNA Prep and sequenced using the MiniSeq platform. Genome assemblies enabled multilocus sequence typing and ospC typing.

Surveillance for Ixodes scapularis and Ixodes pacificus ticks and their associated pathogens in Canada, 2020.

Background: Ixodes scapularis and Ixodes pacificus ticks are the principal vectors of the agent of Lyme disease and several other tick-borne diseases in Canada. Tick surveillance data can be used to identify local tick-borne disease risk areas and direct public health interventions. The objective of this article is to describe the seasonal and spatial characteristics of the main Lyme disease vectors in Canada, and the tick-borne pathogens they carry, using passive and active surveillance data from 2020. Methods: Passive and active surveillance data were compiled from the National Microbiology Laboratory Branch (Public Health Agency of Canada), provincial and local public health authorities, and eTick (an online, image-based platform). Seasonal and spatial analyses of ticks and their associated pathogens are presented, including infection prevalence estimates. Results: In passive surveillance, I. scapularis (n=7,534) were submitted from all provinces except Manitoba and British Columbia, while I. pacificus (n=718) were submitted only from British Columbia. No ticks were submitted from the Territories. The seasonal distribution of I. scapularis submissions was bimodal, but unimodal for I. pacificus. Four tick-borne pathogens were identified in I. scapularis (Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti and Borrelia miyamotoi) and one in I. pacificus (B. miyamotoi). In active surveillance, I. scapularis (n=688) were collected in Ontario, Québec and New Brunswick. Five tick-borne pathogens were identified: B. burgdorferi, A. phagocytophilum, B. microti, B. miyamotoi and Powassan virus. Conclusion: This article provides a snapshot of the distribution of I. scapularis and I. pacificus and their associated human pathogens in Canada in 2020, which can help assess the risk of exposure to tick-borne pathogens in different provinces.

British Columbia's Experience after Implementation of the Treponema pallidum Reverse Algorithm and PCR Detection, 2015 to 2020.

British Columbia (BC) implemented the syphilis reverse screening algorithm and Treponema pallidum PCR testing in 2014. We summarize the performance characteristics of the algorithm, together with PCR direct detection, and report on syphilis cases identified from 2015 to 2020. Prior to 2015, samples for syphilis diagnosis were first screened by rapid plasma reagin (RPR). As of 2015, sera were screened by the Siemens Advia Centaur syphilis assay (enzyme immunoassay [EIA]). Positive and equivocal samples were reflex tested by a T. pallidum passive particle agglutination assay (TPPA) and RPR. We used T. pallidum DNA PCR on clinical samples and restriction fragment length polymorphism analysis to identify azithromycin resistance mutations. Case/epidemiological data were obtained from the BC surveillance system. Of 1,631,519 samples screened by the EIA, 72,492 (4.4%) were positive and 187 (<0.1%) were equivocal. Of EIA-positive/equivocal samples, 10.6% were false positive, and false positivity was higher at lower EIA indices. The reverse algorithm detected 4,693 late latent syphilis cases that likely would have been missed by RPR screening. PCR had a very high sensitivity of 100% versus 52.9% and 52.4% for dark-field (DF) and immunofluorescence (IF) microscopy, respectively. The azithromycin resistance mutation A2058G was identified in 96% of PCR-positive samples, and A2059G was identified in 4%. Annually, there were 944 to 1,467 syphilis cases, with 62% in men who reported male sexual partners. The reverse algorithm had a low false-positive rate and very few equivocal screening results but did identify previously undiagnosed late latent syphilis cases. PCR was more sensitive than both DF and IF microscopy for direct diagnosis and enabled monitoring for azithromycin resistance. IMPORTANCE In this study, we summarize the performance characteristics of the algorithm, together with PCR direct detection and epidemiological analysis, and report on syphilis cases identified from 2015 to 2020. This allowed us to paint a complete picture of the outcome of the utilization of the reverse algorithm for diagnosing syphilis cases. The study clearly showed that the reverse algorithm had a low false-positive rate and very few equivocal screening results but did identify previously undiagnosed late latent syphilis cases. PCR was more sensitive than both DF and IF microscopy for direct diagnosis and enabled monitoring for azithromycin resistance.

Surveillance for Ixodes scapularis and Ixodes pacificus ticks and their associated pathogens in Canada, 2019.

Background: The primary vectors of the agent of Lyme disease in Canada are Ixodes scapularis and Ixodes pacificus ticks. Surveillance for ticks and the pathogens they can transmit can inform local tick-borne disease risk and guide public health interventions. The objective of this article is to characterize passive and active surveillance of the main Lyme disease tick vectors in Canada in 2019 and the tick-borne pathogens they carry. Methods: Passive surveillance data were compiled from the National Microbiology Laboratory Branch and provincial public health data sources. Active surveillance was conducted in selected sentinel sites in all provinces. Descriptive analysis of ticks submitted and infection prevalence of tick-borne pathogens are presented. Seasonal and spatial trends are also described. Results: In passive surveillance, specimens of I. scapularis (n=9,858) were submitted from all provinces except British Columbia and I. pacificus (n=691) were submitted in British Columbia and Alberta. No ticks were submitted from the territories. The seasonal distribution pattern was bimodal for I. scapularis adults, but unimodal for I. pacificus adults. Borrelia burgdorferi was the most prevalent pathogen in I. scapularis (18.8%) and I. pacificus (0.3%). In active surveillance, B. burgdorferi was identified in 26.2% of I. scapularis; Anaplasma phagocytophilum in 3.4% of I. scapularis, and Borrelia miyamotoi and Powassan virus in 0.5% or fewer of I. scapularis. These same tick-borne pathogens were not found in the small number of I. pacificus tested. Conclusion: This surveillance article provides a snapshot of the main Lyme disease vectors in Canada and their associated pathogens, which can be used to monitor emerging risk areas for exposure to tick-borne pathogens.

Performance comparison of micro-neutralization assays based on surrogate SARS-CoV-2 and WT SARS-CoV-2 in assessing virus-neutralizing capacity of anti-SARS-CoV-2 antibodies.

We compared neutralization assays using either the wild-type severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus or surrogate neutralization markers, using characterized sera. We found the results of the neutralization assays 75 % concordant overall and 80 % concordant for samples with high antibody levels. This demonstrates that commercial surrogate SARS-CoV-2 assays offer the potential to assess anti-SARS-CoV-2 antibodies' neutralizing capacity outside CL-3 laboratory containment.

Global phylogeny of Treponema pallidum lineages reveals recent expansion and spread of contemporary syphilis.

Syphilis, which is caused by the sexually transmitted bacterium Treponema pallidum subsp. pallidum, has an estimated 6.3 million cases worldwide per annum. In the past ten years, the incidence of syphilis has increased by more than 150% in some high-income countries, but the evolution and epidemiology of the epidemic are poorly understood. To characterize the global population structure of T. pallidum, we assembled a geographically and temporally diverse collection of 726 genomes from 626 clinical and 100 laboratory samples collected in 23 countries. We applied phylogenetic analyses and clustering, and found that the global syphilis population comprises just two deeply branching lineages, Nichols and SS14. Both lineages are currently circulating in 12 of the 23 countries sampled. We subdivided T. p. pallidum into 17 distinct sublineages to provide further phylodynamic resolution. Importantly, two Nichols sublineages have expanded clonally across 9 countries contemporaneously with SS14. Moreover, pairwise genome analyses revealed examples of isolates collected within the last 20 years from 14 different countries that had genetically identical core genomes, which might indicate frequent exchange through international transmission. It is striking that most samples collected before 1983 are phylogenetically distinct from more recently isolated sublineages. Using Bayesian temporal analysis, we detected a population bottleneck occurring during the late 1990s, followed by rapid population expansion in the 2000s that was driven by the dominant T. pallidum sublineages circulating today. This expansion may be linked to changing epidemiology, immune evasion or fitness under antimicrobial selection pressure, since many of the contemporary syphilis lineages we have characterized are resistant to macrolides.

Passive Tick Surveillance and Detection of Borrelia Species in Ticks from British Columbia, Canada: 2002-2018.

Lyme disease, caused by Borrelia burgdorferi sensu lato (s.l.) complex, is the most common vector-borne disease in North America. This disease has a much lower incidence in western compared with eastern North America. Passive tick surveillance data submitted over 17 years from 2002 to 2018 were analyzed to determine the occurrence of tick species and the prevalence of Borrelia spp. in ticks in British Columbia (BC), Canada. The BC Centre for Disease Control Public Health Laboratory received tick submissions from physicians, veterinarians, and BC residents. Ticks were identified to species, and all ticks, except Dermacentor andersoni, were tested using generic B. burgdorferi s.l. primer sets and species-specific PCR primer sets for B. burgdorferi sensu stricto (s.s.). Tick submission data were analyzed to assess temporal and geographical trends, tick life stages, and tick species. Poisson regression was used to assess temporal trends in annual tick submissions. A total of 15,464 ticks were submitted. Among these, 0.29% (n = 10,235) of Ixodes spp. ticks and 5.3% (n = 434) of Rhipicephalus sanguineus ticks were found carrying B. burgdorferi s.s. B. burgdorferi s.s. was primarily detected in Ixodes pacificus (52%; n = 16) and Ixodes angustus ticks (19%; n = 6) retrieved from humans (n = 5) and animals (n = 26). B. burgdorferi was found in ticks submitted throughout the year. Ixodes spp. ticks were primarily submitted from the coastal regions of southwestern BC, and D. andersoni ticks were primarily submitted from southern interior BC. The number of human tick submissions increased significantly (p < 0.001) between 2013 and 2018. The annual prevalence of B. burgdorferi in ticks remained stable during the study period. These findings correspond to those observed in US Pacific Northwestern states. Passive tick surveillance is an efficient tool to monitor long-term trends in tick distribution and B. burgdorferi prevalence in a low endemicity region.

Comparative Analysis of Capillary vs Venous Blood for Serologic Detection of SARS-CoV-2 Antibodies by RPOC Lateral Flow Tests.

A comparison of rapid point-of-care serology tests using finger prick and venous blood was done on 278 participants. In a laboratory setting, immunoglobulin G (IgG) sensitivity neared 100%; however, IgG sensitivity dramatically dropped (82%) in field testing. Possible factors include finger prick volume variability, hemolysis, cassette readability, and operator training.

Ecology and Epidemiology of Tickborne Pathogens, Washington, USA, 2011-2016.

Tickborne diseases are rare in Washington, USA, and the ecology of these pathogens is poorly understood. We integrated surveillance data from humans and ticks to better describe their epidemiology and ecology. During 2011-2016, a total of 202 tickborne disease cases were reported in Washington residents. Of these, 68 (34%) were autochthonous, including cases of Lyme disease, Rocky Mountain spotted fever, tickborne relapsing fever, and tularemia. During May 2011-December 2016, we collected 977 host-seeking ticks, including Ixodes pacificus, I. angustus, I. spinipalpis, I. auritulus, Dermacentor andersoni, and D. variabilis ticks. The prevalence of Borrelia burgdorferi sensu stricto in I. pacificus ticks was 4.0%; of B. burgdorferi sensu lato, 3.8%; of B. miyamotoi, 4.4%; and of Anaplasma phagocytophilum, 1.9%. We did not detect Rickettsia rickettsii in either Dermacentor species. Case-patient histories and detection of pathogens in field-collected ticks indicate that several tickborne pathogens are endemic to Washington.

Detection of 'Candidatus Ehrlichia khabarensis' in rodents and ticks removed from rodents in British Columbia, Canada.

'Candidatus Ehrlichia khabarensis' was first described from rodents and insectivores in the Far East territory of Khabarovsk on the Russian Pacific Coast. Here we report the detection of DNA from this microorganism in rodents and fed ticks collected from rodents in British Columbia, Canada in 2013-2014. 'Candidatus Ehrlichia khabarensis' was detected in (i) a female Ixodes angustus tick collected from a Peromyscus maniculatus; (ii) a female Dermacentor andersoni tick collected from a Perognathus parvus; (iii) a pool of 2 larval Ixodes pacificus ticks collected from a single P. maniculatus; and (iv) a pool of 3 nymphal I. pacificus ticks collected from a single P. maniculatus. Three of these four rodents (2 P. maniculatus and 1 P. parvus) with infected ticks also had evidence of 'Candidatus Ehrlichia khabarensis' in at least one tissue type. The infected P. maniculatus and Ixodes ticks came from the Vancouver area in western British Columbia and the P. parvus and Dermacentor tick from an inland site in central British Columbia. Although it remains to be determined whether 'Candidatus Ehrlichia khabarensis' has any negative impacts on wildlife, domestic animals or humans, we note that all three tick species found to contain the DNA of this microorganism are known to bite humans. Future detection of this microorganism either in ticks collected from rodents and allowed to molt to the next life stage prior to being tested, or from host-seeking ticks, is required to determine if it can survive the tick's molt after being ingested via an infectious blood meal.

Increased Discrimination of Treponema pallidum Strains by Subtyping With a 4-Component System Incorporating a Mononucleotide Tandem Repeat in rpsA.

A guanine mononucleotide repeat in the rpsA (tp0279) gene was evaluated for improved strain discrimination using 72 Treponema pallidum-positive specimens. The tandem repeat combined with the enhanced Centers for Disease Control and Prevention typing system resulted in increased discrimination and should be useful for molecular epidemiologic studies on syphilis especially in outbreaks and among men who have sex with men.

A Retrospective Cohort Study of Tick Paralysis in British Columbia.

BACKGROUND: Tick paralysis is a frequently overlooked severe disease characterized by bilateral ascending flaccid paralysis caused by a neurotoxin produced by feeding ticks. We aimed to characterize suspected tick paralysis cases documented at the BC Centre for Disease Control (BCCDC) in British Columbia (BC) from 1993 to 2016 and reviewed prevention, diagnosis, and treatment considerations. METHODS: Demographic, geographic, and clinical data from test requisition forms for ticks submitted to the BCCDC Public Health Laboratory (PHL) from patients across BC between 1993 and 2016 for suspected human and animal tick paralysis were reviewed. Descriptive statistics were generated to characterize tick paralysis cases in BC, including tick species implicated, seasonality of disease, and regional differences. RESULTS: From 1993 to 2016, there were 56 cases of suspected tick paralysis with at least one tick specimen submitted for testing at the BCCDC PHL. Humans and animals were involved in 43% and 57% of cases, respectively. The majority of cases involved a Dermacentor andersoni tick (48 cases or 86%) and occurred between the months of April and June (49 cases or 88%). Among known locations of tick acquisition, the Interior region of BC was disproportionately affected, with 25 cases (69%) of tick bites occurring in that area. CONCLUSIONS: Tick paralysis is a rare condition in BC. The region of highest risk is the Interior, particularly during the spring and summer months. Increasing awareness of tick paralysis among healthcare workers and the general public is paramount to preventing morbidity and mortality from this rare disease.

Surveillance for Borrelia burgdorferi in Ixodes Ticks and Small Rodents in British Columbia.

To determine the prevalence of Borrelia burgdorferi in British Columbian ticks, fieldwork was conducted over a 2-year period. In all, 893 ticks (Ixodes pacificus, I. angustus, I. soricis, Ixodes spp., and Dermacentor andersoni) of different life stages were retrieved from 483 small rodents (Peromyscus maniculatus, Perognathus parvus, and Reithrodontomys megalotis). B. burgdorferi DNA was detected in 5 out of 359 tick pools, and 41 out of 483 mice were serologically confirmed to have antibodies against B. burgdorferi. These results were consistent with previous studies, data from passive surveillance in British Columbia, and data from neighboring states in the Pacific Northwest, suggesting a continually low prevalence of B. burgdorferi in British Columbia ticks.

A decade of experience: Cryptococcus gattii in British Columbia.

It has been over a decade since Cryptococcus gattii was first recognized as the causative organism of an outbreak of cryptococcosis on Vancouver Island, British Columbia. A number of novel observations have been associated with the study of this emergent pathogen. A novel genotype of C. gattii, VGIIa was described as the major genotype associated with clinical disease. Minor genotypes, VGIIb and VGI, are also responsible for disease in British Columbians, in both human and animal populations. The clinical major genotype VGIIa and minor genotype VGIIb are identical to C. gattii isolated from the environment of Vancouver Island. There is more heterogeneity in VGI, and a clear association with the environment is not apparent. Between 1999 and 2010, there have been 281 cases of C. gattii cryptococcosis. Risk factors for infection are reported to be age greater than 50 years, history of smoking, corticosteroid use, HIV infection, and history of cancer or chronic lung disease. The major C. gattii genotype VGIIa is as virulent in mice as the model Cryptococcus, H99 C. neoformans, although the outbreak strain produces a less protective inflammatory response in C57BL/6 mice. The minor genotype VGIIb is significantly less virulent in mouse models. Cryptococcus gattii is found associated with native trees and soil on Vancouver Island. Transiently positive isolations have been made from air and water. An ecological niche for this organism is associated within a limited biogeoclimatic zone characterized by daily average winter temperatures above freezing.

Detection of Lyme disease spirochete, Borrelia burgdorferi sensu lato, including three novel genotypes in ticks (Acari: Ixodidae) collected from songbirds (Passeriformes) across Canada.

Lyme disease is reported across Canada, but pinpointing the source of infection has been problematic. In this three-year, bird-tick-pathogen study (2004-2006), 366 ticks representing 12 species were collected from 151 songbirds (31 passerine species/subspecies) at 16 locations Canada-wide. Of the 167 ticks/pools tested, 19 (11.4%) were infected with Borrelia burgdorferi sensu lato (s.l.). Sequencing of the rrf-rrl intergenic spacer gene revealed four Borrelia genotypes: B. burgdorferi sensu stricto (s.s.) and three novel genotypes (BC genotype 1, BC genotype 2, BC genotype 3). All four genotypes were detected in spirochete-infected Ixodes auritulus (females, nymphs, larvae) suggesting this tick species is a vector for B. burgdorferi s.l. We provide first-time records for: ticks in the Yukon (north of 60 degrees latitude), northernmost collection of Amblyomma americanum in North America, and Amblyomma imitator in Canada. First reports of bird-derived ticks infected with B. burgdorferi s.l. include: live culture of spirochetes from Ixodes pacificus (nymph) plus detection in I. auritulus nymphs, Ixodes scapularis in New Brunswick, and an I. scapularis larva in Canada. We provide the first account of B. burgdorferi s. l. in an Ixodes muris tick collected from a songbird anywhere. Congruent with previous data for the American Robin, we suggest that the Common Yellowthroat, Golden-crowned Sparrow, Song Sparrow, and Swainson's Thrush are reservoir-competent hosts. Song Sparrows, the predominant hosts, were parasitized by I. auritulus harboring all four Borrelia genotypes. Our results show that songbirds import B. burgdorferi s.l.-infected ticks into Canada. Bird-feeding I. scapularis subadults were infected with Lyme spirochetes during both spring and fall migration in eastern Canada. Because songbirds disperse millions of infected ticks across Canada, people and domestic animals contract Lyme disease outside of the known and expected range.

Rapid introduction of Lyme disease spirochete, Borrelia burgdorferi sensu stricto, in Ixodes scapularis (Acari: Ixodidae) established at Turkey Point Provincial Park, Ontario, Canada.

Borrelia burgdorferi sensu stricto (s.s.) was isolated from questing adult Ixodes scapularis Say ticks collected from Turkey Point Provincial Park (TPPP), Ontario, Canada during 2005-2006. DNA from ten (67%) of 15 pools of ticks was confirmed positive for B. burgdorferi s.s. using polymerase chain reaction (PCR) by targeting the rrf (5S)-rrl (23S) intergenic spacer region and OspA genes. This significant infection rate indicates an accelerated development of B. burgdorferi s.s. in TPPP, because this pathogen was not detected five years previously during sampling of the three motile life stages of I. scapularis. Our study provides the initial report of the presence of B. burgdorferi s.s. in TPPP, which is now endemic for Lyme disease. Ultimately, people and domestic animals are at risk of contracting Lyme disease when they frequent this park.

Neurosyphilitic gumma in a homosexual man with HIV infection confirmed by polymerase chain reaction.

The brain gumma is a rare manifestation of the tertiary stage of syphilis. A case of neurosyphilitic gumma was confirmed by the Treponema pallidum polymerase chain reaction in a 46-year-old HIV-positive homosexual man. The patient presented with a severe headache and was hospitalized. A computed tomography scan was performed which revealed a left frontal lobe mass. Lymphoma was suspected. However, infectious disease diagnostics were performed on the cerebrospinal fluid that included investigations for syphilis and other microbiological agents such as Toxoplasma gondii. This revealed a reactive venereal disease research laboratory test, a reactive syphilis rapid plasma reagin and a reactive T. pallidum particle agglutination test. The patient was treated for syphilis till complete recovery.

Molecular methods used in clinical laboratory: prospects and pitfalls.

The role of molecular detection, identification and typing or fingerprinting of microorganisms has shifted gradually from the academic world to the routine diagnostic laboratory. Molecular methods have been used increasingly over the past decade to improve the sensitivity, specificity and turn-around time in the clinical laboratory. Molecular methods have also been used to identify new and nonculturable agents. Many high-throughput molecular tests are now available commercially, which impacts on the infrastructure in many of the diagnostic laboratories. In this paper, we take an overall look at the use of molecular methods (prospects vs. pitfalls) based on our clinical and public health experience, particularly as they related to Borrelia burgdorferi, a vector-borne pathogen, Treponema pallidum, a re-emerging sexually transmitted global pathogen, and West Nile virus, a newly recognized virus in North America.