High Seroprevalence of Jamestown Canyon Virus among Deer and Humans, Nova Scotia, Canada.

Using residual serum samples from Nova Scotia, Canada, we found that 87.8% of tested deer and an estimated 20.6% of the human population were infected with Jamestown Canyon virus. Human seropositivity reached 48.2% in 1 region. This virus may be an underrecognized cause of disease in Nova Scotia.

Apoptosis, autophagy and unfolded protein response pathways in Arbovirus replication and pathogenesis.

Arboviruses are pathogens that widely affect the health of people in different communities around the world. Recently, a few successful approaches toward production of effective vaccines against some of these pathogens have been developed, but treatment and prevention of the resulting diseases remain a major health and research concern. The arbovirus infection and replication processes are complex, and many factors are involved in their regulation. Apoptosis, autophagy and the unfolded protein response (UPR) are three mechanisms that are involved in pathogenesis of many viruses. In this review, we focus on the importance of these pathways in the arbovirus replication and infection processes. We provide a brief introduction on how apoptosis, autophagy and the UPR are initiated and regulated, and then discuss the involvement of these pathways in regulation of arbovirus pathogenesis.

Seroprevalence of Babesia microti infection in Canadian blood donors.

BACKGROUND: Human babesiosis, caused by the intraerythrocytic protozoan parasite Babesia microti, is primarily transmitted by tick bites and is also transmitted by transfusion. Infections have been identified in U.S. blood donors close to Canadian borders. We aimed to assess the risk of transfusion-transmitted babesiosis in Canada by examining infections in ticks and seroprevalence in blood donors. STUDY DESIGN AND METHODS: Passive surveillance (receipt of ticks submitted by the public) was used to identify regions for tick drag sampling (active surveillance, 2009-2014). All ticks were tested for B. microti using an indirect immunofluorescent antibody assay (Imugen, Inc.). Between July and December 2013, blood donations from selected sites (southern Manitoba, Ontario, Québec, New Brunswick, and Nova Scotia) near endemic U.S. regions were tested for antibody to B. microti. Donors completed a questionnaire about risk travel and possible tick exposure. RESULTS: Of approximately 12,000 ticks submitted, 14 were B. microti positive (10 in Manitoba, one in Ontario, one in Québec, two in New Brunswick). From active tick surveillance, six of 361 ticks in Manitoba were positive (1.7%), three of 641 (0.5%) in Québec, and none elsewhere. There were 26,260 donors at the selected sites of whom 13,993 (53%) were tested. None were positive for antibody to B. microti. In 2013, 47% of donors visited forested areas in Canada, and 41% traveled to the United States. CONCLUSION: The data do not suggest that laboratory-based testing is warranted at this time. However, there are indicators that B. microti may be advancing into Canada and ongoing monitoring of tick populations and donor seroprevalence is indicated.

Epidemiology of Lyme Disease, Nova Scotia, Canada, 2002-2013.

Ixodes scapularis ticks, which transmit Borrelia burgdorferi, the causative agent of Lyme disease (LD), are endemic to at least 6 regions of Nova Scotia, Canada. To assess the epidemiology and prevalence of LD in Nova Scotia, we analyzed data from 329 persons with LD reported in Nova Scotia during 2002-2013. Most patients reported symptoms of early localized infection with rash (89.7%), influenza-like illness (69.6%), or both; clinician-diagnosed erythema migrans was documented for 53.2%. In a separate serosurvey, of 1,855 serum samples screened for antibodies to B. burgdorferi, 2 were borderline positive (both with an indeterminate IgG on Western blot), resulting in an estimated seroprevalence of 0.14% (95% CI 0.02%-0.51%). Although LD incidence in Nova Scotia has risen sharply since 2002 and is the highest in Canada (16/100,000 population in 2013), the estimated number of residents with evidence of infection is low, and risk is localized to currently identified LD-endemic regions.

A comparison of sampling and testing approaches for the surveillance of SARS-CoV-2 in farmed American mink.

Surveillance for SARS-CoV-2 in American mink (Neovison vison) is a global priority because outbreaks on mink farms have potential consequences for animal and public health. Surveillance programs often focus on screening natural mortalities; however, significant knowledge gaps remain regarding sampling and testing approaches. Using 76 mink from 3 naturally infected farms in British Columbia, Canada, we compared the performance of 2 reverse-transcription real-time PCR (RT-rtPCR) targets (the envelope [E] and RNA-dependent RNA polymerase [RdRp] genes) as well as serology. We also compared RT-rtPCR and sequencing results from nasopharyngeal, oropharyngeal, skin, and rectal swabs, as well as nasopharyngeal samples collected using swabs and interdental brushes. We found that infected mink were generally RT-rtPCR-positive on all samples; however, Ct values differed significantly among sample types (nasopharyngeal < oropharyngeal < skin < rectal). There was no difference in the results of nasopharyngeal samples collected using swabs or interdental brushes. For most mink (89.4%), qualitative (i.e., positive vs. negative) serology and RT-rtPCR results were concordant. However, mink were positive on RT-rtPCR and negative on serology and vice versa, and there was no significant correlation between Ct values on RT-rtPCR and percent inhibition on serology. Both the E and RdRp targets were detectable in all sample types, albeit with a small difference in Ct values. Although SARS-CoV-2 RNA can be detected in multiple sample types, passive surveillance programs in mink should focus on multiple target RT-rtPCR testing of nasopharyngeal samples in combination with serology.

Modified Two-Tiered Testing Enzyme Immunoassay Algorithm for Serologic Diagnosis of Lyme Disease.

The modified 2-tier testing algorithm (MTTT) for Lyme disease (LD) has been approved by the US Food and Drug Administration. In this study, we show that the MTTT detected 28% more cases of early infection compared with the standard 2-tier algorithm while retaining high specificity in a region with a high incidence of LD.

Serologic testing for Bartonella in Manitoba, Canada, 2010-2020: a retrospective case series.

BACKGROUND: Bartonella are gram-negative bacilli not identified by routine bacterial culture. The objectives of this study were to review the results of all serologic testing for Bartonella ordered in Manitoba, Canada, and to review cases with positive test results among adults to assess species identification, risk factors, clinical manifestations and outcomes. METHODS: This retrospective study included all Bartonella serologic tests ordered in Manitoba and performed at the National Microbiology Laboratory, Winnipeg, from Jan. 1, 2010, until Dec. 31, 2020. We analyzed the aggregate data for all serologic tests for Bartonella for patients of all ages. We reviewed the charts of adult (age ≥ 18 yr) patients with serologic positivity for Bartonella who had a medical chart at 1 of Winnipeg's 2 largest hospitals (Health Sciences Centre and St. Boniface Hospital) to extract clinical and demographic data and create a case series. Descriptive statistics were performed. RESULTS: During the study period, 1014 Bartonella serologic tests were ordered in adult and pediatric patients, of which 24 (2.4%) gave a positive result. Sixteen adults (12 men and 4 women; mean age 48 yr) seen at a participating hospital had a positive result. Molecular species-level identification occurred on explanted cardiac valves in 5 (31%) of the 16 cases; B. quintana was identified in all 5. Six patients (38%) were diagnosed with probable B. quintana infection, for a total of 11 B. quintana cases (69%); 8 (73%) of the 11 had endocarditis. Four cases of B. quintana infection (36%) were associated with rural residence. Four cases (25%) of probable B. henselae were identified; 2 patients had fever and lymphadenopathy, and 2 had endocarditis. The remaining patient was deemed to have a false-positive result as his B. henselae titre was at the threshold for positivity, his B. quintana serologic test gave a negative result, and his clinical syndrome was not suggestive of Bartonella infection. Two patients died; both had multivalvular B. quintana endocarditis with ruptured intracranial mycotic aneurysms. INTERPRETATION: Bartonella quintana was a common cause of Bartonella serologic positivity among adults in Manitoba in 2010-2020 and was associated with endocarditis and systemic embolization. As B. quintana is transmitted by body lice, active case finding for people who lack suitable housing, both in urban and rural settings, should prioritize those with elevated Bartonella titres to receive echocardiography and detect endocarditis before systemic embolization occurs.

Mosquitoes Know No Borders: Surveillance of Potential Introduction of Aedes Species in Southern Québec, Canada.

Current climatic conditions limit the distribution of Aedes (Stegomyia) albopictus (Skuse, Diptera: Culicidae) in the north, but predictive climate models suggest this species could establish itself in southern Canada by 2040. A vector of chikungunya, dengue, yellow fever, Zika and West Nile viruses, the Ae. Albopictus has been detected in Windsor, Ontario since 2016. Given the potential public health implications, and knowing that Aedes spp. can easily be introduced by ground transportation, this study aimed to determine if specimens could be detected, using an adequate methodology, in southern Québec. Mosquitoes were sampled in 2016 and 2017 along the main roads connecting Canada and the U.S., using Biogent traps (Sentinel-2, Gravide Aedes traps) and ovitraps. Overall, 24 mosquito spp. were captured, excluding Ae. Albopictus, but detecting one Aedes (Stegomyia) aegypti (Skuse) specimen (laid eggs). The most frequent species among captured adults were Ochlerotatus triseriatus, Culex pipiens complex, and Ochlerotatus japonicus (31.0%, 26.0%, and 17.3%, respectively). The present study adds to the increasing number of studies reporting on the range expansions of these mosquito species, and suggests that ongoing monitoring, using multiple capture techniques targeting a wide range of species, may provide useful information to public health with respect to the growing risk of emerging mosquito-borne diseases in southern Canada.

Bird Species Involved in West Nile Virus Epidemiological Cycle in Southern Québec.

Despite many studies on West Nile Virus (WNV) in the US, including the reservoir role of bird species and the summer shifts of the Culex mosquito, feeding from birds to mammals, there have been few equivalent studies in the neighboring regions of Canada where WNV is endemic. Here, a priority list of bird species likely involved in WNV transmission in the greater Montréal area is constructed by combining three sources of data: (i) from WNV surveillance in wild birds (2002-2015); (ii) blood meal analysis of Culex pipiens-restuans (CPR), the primary enzootic vectors of WNV in the region, collected from surveillance in 2008 and 2014; (iii) literature review on the sero-prevalence/host competence of resident birds. Each of these data sources yielded 18, 23 and 53 species, and overall, 67 different bird species were identified as potential WNV amplifiers/reservoirs. Of those identified from CPR blood meals, Common starlings, American robins, Song sparrows and House sparrows ranked the highest and blood meal analysis demonstrated a seasonal shift in feed preference from birds to mammals by CPR. Our study indicates that there are broad similarities in the ecology of WNV between our region and the northeastern US, although the relative importance of bird species varies somewhat between regions.

Paraparesis in a polar bear (Ursus maritimus) associated with West Nile virus infection.

A polar bear (Ursus maritimus) housed at the Toronto Zoo presented with acute-onset, nonambulatory paraparesis. Physical examination 24 hr after onset was otherwise unremarkable, spinal radiographs looked normal, and blood tests indicated mild dehydration. With continued deterioration in its general condition, euthanasia was elected a day later. Necropsy did not reveal a cause for the major presenting clinical signs. Serum collected at the time of initial examination was positive for West Nile virus (WNV) antibodies in a serum neutralization assay and at the time of euthanasia was positive in both a competitive enzyme-linked immunosorbent assay and in a plaque reduction neutralization assay. The major microscopic finding was a mild-to-moderate nonsuppurative meningoencephalomyelitis. WNV was not detected by immunohistochemistry in brain or spinal cord or by real-time reverse transcription-polymerase chain reaction (RT-PCR) and cell culture of brain and kidney, but it was isolated and identified by RT-PCR in second passage cell culture of spleen. Retrospective immunohistochemistry on spleen revealed rare antigen-positive cells, probably macrophages. Prevention of exposure to potentially WNV-infected mosquitoes or vaccination of captive bears against WNV should be considered.

Tick infestations of wildlife and companion animals in Ontario, Canada, with detection of human pathogens in Ixodes scapularis ticks.

The growing risk of transmission of tick-borne zoonotic pathogens to humans in Ontario, Canada, warrants investigations into regional tick distribution, tick burdens of local peridomestic animals, and prevalence of tick-borne pathogens. The objectives of this study were to investigate the geographic distribution and magnitude of tick infestations in opportunistically sampled mammalian wildlife and companion animals (i.e., dogs) in southern Ontario and to test these ticks for evidence of zoonotic tick-borne pathogens. Ticks collected from wildlife carcasses, live-trapped wildlife and companion animals (2015-2016), as well as wildlife diagnostic cases (2011-2013), were identified to species and life stage. Ixodes scapularis ticks were tested by real-time PCR for Anaplasma phagocytophilum, Babesia microti, Borrelia miyamotoi and Borrelia burgdorferi sensu stricto (s.s.). Amblyomma americanum ticks were tested for Ehrlichia chaffeensis. A total of 1687 ticks of six species were collected from 334 animals, including 224 raccoons (n = 1381 ticks) and 50 dogs (n = 67 ticks). The most common tick species collected from parasitized raccoons were Ixodes texanus (n = 666 ticks) and Dermacentor variabilis (n = 600 ticks), which were removed from 58.5% (median: 2 ticks; range: 1-36) and 49.1% (median: 2 ticks; range: 1-64) of raccoons, respectively. Of I. scapularis tested, 9.3% (4/43) were positive for Bo. burgdorferi s.s. and 2.3% (1/43) for A. phagocytophilum. These results reveal that numerous tick species parasitize common, peridomestic wildlife and that at least two zoonotic, tick-borne pathogens circulate in southern Ontario. Host-tick vector-pathogen dynamics should continue to be monitored in the face of global climate change, landscape alterations and expanding human populations.

Detection of municipalities at-risk of Lyme disease using passive surveillance of Ixodes scapularis as an early signal: A province-specific indicator in Canada.

Lyme disease, the most commonly reported vector-borne disease in North America, is caused by the spirochete Borrelia burgdorferi sensu stricto, which is transmitted by Ixodes scapularis in eastern Canada and Ixodes pacificus in western Canada. Recently, the northward range expansion of I. scapularis ticks, in south-eastern Canada, has resulted in a dramatic increase in the incidence of human Lyme disease. Detecting emerging areas of Lyme disease risk allows public health to target disease prevention efforts. We analysed passive tick surveillance data from Ontario and Manitoba to i) assess the relationship between the total numbers of I. scapularis submissions in passive surveillance from humans, and the number of human Lyme disease cases, and ii) develop province-specific acarological indicators of risk that can be used to generate surveillance-based risk maps. We also assessed associations between numbers of nymphal I. scapularis tick submissions only and Lyme disease case incidence. Using General Estimating Equation regression, the relationship between I. scapularis submissions (total numbers and numbers of nymphs only) in each census sub-division (CSD) and the number of reported Lyme disease cases was positively correlated and highly significant in the two provinces (P ≤ 0.001). The numbers of I. scapularis submissions over five years discriminated CSDs with ≥ 3 Lyme disease cases from those with < 3 cases with high accuracy when using total numbers of tick submission (Receiver Operating Characteristics area under the curve [AUC] = 0.89) and moderate accuracy (AUC = 0.78) when using nymphal tick submissions only. In Ontario the optimal cut-off point was a total 12 tick submissions from a CSD over five years (Sensitivity = 0.82, Specificity = 0.84), while in Manitoba the cut-off point was five ticks (Sensitivity = 0.71, Specificity = 0.79) suggesting regional variability of the risk of acquiring Lyme disease from an I. scapularis bite. The performances of the acarological indicators developed in this study for Ontario and Manitoba support the ability of passive tick surveillance to provide an early signal of the existence Lyme disease risk areas in regions where ticks and the pathogens they transmit are expanding their range.

RVFV Infection in Goats by Different Routes of Inoculation.

Rift Valley fever virus (RVFV) is a zoonotic arbovirus of the Phenuiviridae family. Infection causes abortions in pregnant animals, high mortality in neonate animals, and mild to severe symptoms in both people and animals. There is currently an ongoing effort to produce safe and efficacious veterinary vaccines against RVFV in livestock to protect against both primary infection in animals and zoonotic infections in people. To test the efficacy of these vaccines, it is essential to have a reliable challenge model in relevant target species, including ruminants. We evaluated two goat breeds (Nubian and LaMancha), three routes of inoculation (intranasal, mosquito-primed subcutaneous, and subcutaneous) using an infectious dose of 107 pfu/mL, a virus strain from the 2006⁻2007 Kenyan/Sudan outbreak and compared the effect of using virus stocks produced in either mammalian or mosquito cells. Our results demonstrated that the highest and longest viremia titers were achieved in Nubian goats. The Nubian breed was also efficient at producing clinical signs, consistent viremia (peak viremia: 1.2 × 10³â»1.0 × 105 pfu/mL serum), nasal and oral shedding of viral RNA (1.5 × 10¹â»8 × 106 genome copies/swab), a systemic infection of tissues, and robust antibody responses regardless of the inoculation route. The Nubian goat breed and a needle-free intranasal inoculation technique could both be utilized in future vaccine and challenge studies. These studies are important for preventing the spread and outbreak of zoonotic viruses like RVFV and are supported by the Canadian-led BSL4ZNet network.

Anaplasmosis: An emerging tick-borne disease of importance in Canada.

Human Granulocytic Anaplasmosis (HGA) is an infection caused by the intracellular bacterium Anaplasma phagocytophilum. As a tick-borne disease, the public health impact of HGA continues to increase with range expansion of the disease vector. The clinical presentation of HGA is often a non-specific febrile illness. The presence of leukopenia, thrombocytopenia, and mild hepatic injury are frequently noted on laboratory investigations, which can be important diagnostic clues in attaining an appropriate diagnosis. Herein we present three cases of HGA, highlighting the spectrum of disease by which HGA can manifest. Although each case has their unique features, we outline important shared clinical elements to facilitate an empiric diagnosis while definitive laboratory investigations are pending. Our case series further serves to highlight the critical importance of prompt antimicrobial treatment to reduce morbidity and potential mortality.

Evidence for increasing densities and geographic ranges of tick species of public health significance other than Ixodes scapularis in Québec, Canada.

Climate change is driving emergence and establishment of Ixodes scapularis, the main vector of Lyme disease in Québec, Canada. As for the black-legged tick, I. scapularis Say, global warming may also favor northward expansion of other species of medically important ticks. The aims of this study were to determine (1) current diversity and abundance of ticks of public health significance other than I. scapularis, (2) sex and age of the human population bitten by these ticks (3), and the seasonal and geographic pattern of their occurrence. From 2007 to 2015, twelve tick species other than I. scapularis were submitted in the Québec passive tick surveillance program. Of these 9243 ticks, 91.2% were Ixodes cookei, 4.1% were Dermacentor variabilis, 4.0% were Rhipicephalus sanguineus and 0.7% were Amblyomma americanum. The combined annual proportion of submitted I. cookei, D. variabilis, R. sanguineus and A. americanum ticks in passive surveillance rose from 6.1% in 2007 to 16.0% in 2015 and an annual growing trend was observed for each tick species. The number of municipalities where I. cookei ticks were acquired rose from 104 to 197 during the same period. Of the 862 people bitten by these ticks, 43.3% were I. cookei ticks removed from children aged < 10 years. These findings demonstrate the need for surveillance of all the tick species of medical importance in Québec, particularly because climate may increase their abundance and geographic ranges, increasing the risk to the public of the diseases they transmit.

Powassan Virus and Other Arthropod-Borne Viruses in Wildlife and Ticks in Ontario, Canada.

Powassan virus (POWV) is a tick-borne zoonosis maintained in natural enzootic cycles between ixodid ticks and wild mammals. Reported human cases have increased in recent years; these infections can be fatal or lead to long-term neurologic sequelae. However, both the geographic distribution and the role of common, potential mammalian hosts in POWV transmission are poorly understood, creating challenges to public health surveillance. We looked for evidence of POWV infection among candidate wildlife host species and ticks collected from mammals and birds in southern Ontario. Tissues (including blood) and ticks from trapped wild mammals were collected in the summers of 2015 and 2016. Ticks removed from dogs in 2015-2016 and wildlife diagnostic cases from 2011 to 2013 were also included. Tissue and tick (Ixodes spp.) homogenates were tested for POWV by reverse transcriptase-polymerase chain reaction (RT-PCR). In addition, sera from wild mammals were tested for antibodies to POWV, West Nile virus (WNV), and heartland virus (HRTV) by plaque reduction neutralization test. All 724 tissue samples were negative for POWV by RT-PCR. One of 53 pools of Ixodes cookei (among 98 total tick pools) was RT-PCR positive for deer tick virus (POWV) lineage. Antibodies to POWV and WNV were detected in 0.4% of 265 and 6.1% of 264 samples, respectively, and all of 219 serum samples tested negative for anti-HRTV antibodies. These results reveal low POWV detection rates in southern Ontario, while highlighting the challenges and need for continued efforts into understanding POWV epidemiology and targeted surveillance strategies.

Seroprevalence of Rift Valley Fever Virus Antibodies in Cattle in Mali, 2005-2014.

Rift Valley fever virus (RVFV) outbreaks have considerable impact on human and animal health. Here, we are reporting a serosurvey of cattle from all regions of Mali. These demonstrated that few had been exposed to RVFV from 2005 to 2014. Recent outbreaks of RVF in Niger and a single human case in Mali provide justification for further entomological and ecological studies of this virus.

DNA vaccination protects mice against Zika virus-induced damage to the testes.

Zika virus (ZIKV) is an emerging pathogen causally associated with serious sequelae in fetuses, inducing fetal microcephaly and other neurodevelopment defects. ZIKV is primarily transmitted by mosquitoes, but can persist in human semen and sperm, and sexual transmission has been documented. Moreover, exposure of type-I interferon knockout mice to ZIKV results in severe damage to the testes, epididymis and sperm. Candidate ZIKV vaccines have shown protective efficacy in preclinical studies carried out in animal models, and several vaccines have entered clinical trials. Here, we report that administration of a synthetic DNA vaccine encoding ZIKV pre-membrane and envelope (prME) completely protects mice against ZIKV-associated damage to the testes and sperm and prevents viral persistence in the testes following challenge with a contemporary strain of ZIKV. These data suggest that DNA vaccination merits further investigation as a potential means to reduce ZIKV persistence in the male reproductive tract.

Use of IgG avidity to indirectly monitor epizootic transmission of sin nombre virus in deer mice (Peromyscus maniculatus).

An IgG avidity assay was developed to differentiate deer mice that had recently acquired Sin Nombre virus (SNV) from those that were infected in the distant past. Using this procedure, low avidity antibodies were predominantly detected in experimentally infected deer mice (89.5%) within the first 30 days post-inoculation. The assay was then applied to sera from naturally infected deer mice collected during a field investigation associated with a cluster of hantavirus pulmonary syndrome cases. A higher proportion of seropositive mice collected during the outbreak had serum with low avidity antibodies (16.7%) when compared with mice trapped four months later (5.7%). Sin Nombre virus RNA was detectable in blood in a similar fraction of low- (45%) and high- (38.7%) avidity groups. Non-adult mice were more likely to contain low-avidity antibodies (44.4%) than were adults (9.6%). Our results indicate that the IgG avidity assay shows promise as a tool to better characterize epizootic intensity and to identify factors involved in SNV transmission.

Evaluation of commercial assays for detecting West Nile virus antigen.

Two commercially available West Nile virus (WNV) detection assays (RAMP WNV test, Response Biomedical Corp., Burnaby, British Columbia, Canada; and VecTest WNV antigen assay, Medical Analysis Systems, Inc., Camarillo, CA) were compared for sensitivity, specificity, and ability to detect WNV in field-collected mosquito pools. Serially diluted stock seed WNV and St. Louis encephalitis virus (SLEV) were used to determine sensitivity and specificity. The RAMP WNV test detected WNV at concentrations as low as 3.17 log10 plaque-forming units per milliliter (PFU/ml), whereas the VecTest assay detected WNV at concentrations as low as 5.17 log10 PFU/ml. Neither test cross-reacted with SLEV. A WNV-specific reverse transcriptase polymerase chain reaction was used to identify positives among field-collected mosquito pools. The RAMP WNV test detected 94% of positive pools and the VecTest assay detected 65% of the positive field-collected pools. Despite these differences, both assays have characteristics that make them useful in WNV surveillance programs.