Assessing the Efficiency of Local Rabies Vaccination Strategies for Raccoons (Procyon lotor) in an Urban Setting.

Raccoon rabies virus (RRV) has been managed using multiple vaccination strategies, including oral rabies vaccination and trap-vaccinate-release (TVR). Identifying a rabies vaccination strategy for an area is a nontrivial task. Vaccination strategies differ in the amount of effort and monetary costs required to achieve a particular level of vaccine seroprevalence (efficiency). Simulating host movement relative to different vaccination strategies in silico can provide a useful tool for exploring the efficiency of different vaccination strategies. We refined a previously developed individual-based model of raccoon movement to evaluate vaccination strategies for urban Hamilton, Ontario, Canada. We combined different oral rabies vaccination baiting (hand baiting, helicopter, and bait stations) with TVR strategies and used GPS data to parameterize and simulate raccoon movement in Hamilton. We developed a total of 560 vaccination strategies, in consultation with the Ontario Ministry of Natural Resources and Forestry, for RRV control in Hamilton. We documented the monetary costs of each vaccination strategy and estimated the population seroprevalence. Intervention costs and seroprevalence estimates were used to calculate the efficiency of each strategy to meet targets set for the purpose of RRV control. Estimated seroprevalence across different strategies varied widely, ranging from less than 5% to more than 70%. Increasing bait densities (distributed using by hand or helicopter) led to negligible increase in seroprevalence. Helicopter baiting was the most efficient and TVR was the least efficient, but helicopter-based strategies led to lower levels of seroprevalence (6-12%) than did TVR-based strategies (17-70%). Our simulations indicated that a mixed strategy including at least some TVR may be the most efficient strategy for a local urban RRV control program when seroprevalence levels >30% may be required. Our simulations provide information regarding the efficiency of different vaccination strategies for raccoon populations, to guide local RRV control in urban settings.

Comparing Control Intervention Scenarios for Raccoon Rabies in Southern Ontario between 2015 and 2025.

The largest outbreak of raccoon rabies in Canada was first reported in Hamilton, Ontario, in 2015 following a probable translocation event from the United States. We used a spatially-explicit agent-based model to evaluate the effectiveness of provincial control programs in an urban-centric outbreak if control interventions were used until 2025, 2020, or never used. Calibration tests suggested that a seroprevalence of protective rabies antibodies 2.1 times higher than that inferred from seroprevalence in program assessments was required in simulations to replicate observed raccoon rabies cases. Our simulation results showed that if control interventions with an adjusted seroprevalence were used until 2025 or 2020, the probability of rabies elimination due to control intervention use was 49.2% and 42.1%, respectively. However, if controls were never used, the probability that initial rabies cases failed to establish a sustained outbreak was only 18.2%. In simulations where rabies was not successfully eliminated, using control interventions until 2025 resulted in 67% fewer new infections compared to only applying controls until 2020 and in 90% fewer new infections compared to no control intervention use. However, the model likely underestimated rabies elimination rates since we did not adjust for adaptive control strategies in response to changes in rabies distributions and case numbers, as well as extending control interventions past 2025. Our agent-based model offers a cost-effective strategy to evaluate approaches to rabies control applications.

Divergent SARS-CoV-2 variant emerges in white-tailed deer with deer-to-human transmission.

Wildlife reservoirs of broad-host-range viruses have the potential to enable evolution of viral variants that can emerge to infect humans. In North America, there is phylogenomic evidence of continual transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) from humans to white-tailed deer (Odocoileus virginianus) through unknown means, but no evidence of transmission from deer to humans. We carried out an observational surveillance study in Ontario, Canada during November and December 2021 (n = 300 deer) and identified a highly divergent lineage of SARS-CoV-2 in white-tailed deer (B.1.641). This lineage is one of the most divergent SARS-CoV-2 lineages identified so far, with 76 mutations (including 37 previously associated with non-human mammalian hosts). From a set of five complete and two partial deer-derived viral genomes we applied phylogenomic, recombination, selection and mutation spectrum analyses, which provided evidence for evolution and transmission in deer and a shared ancestry with mink-derived virus. Our analysis also revealed an epidemiologically linked human infection. Taken together, our findings provide evidence for sustained evolution of SARS-CoV-2 in white-tailed deer and of deer-to-human transmission.

Natural disease and evolution of an Amdoparvovirus endemic in striped skunks (Mephitis mephitis).

Striped skunks (Mephitis mephitis) densely populate the human-animal interface of suburbia throughout North America. Skunks share that habitat with numerous related mesocarnivores, where increased contact, competition for shared food and water sources and other stressors contribute to increased exposure and susceptibility to viral infection. The recently identified skunk amdoparvovirus (SKAV) has been detected at high prevalence in skunks and occasionally in mink, but its distribution in North America is unknown. To understand the impact of SKAV in striped skunks and the risk posed to related species, we investigated the geographic distribution of SKAV, analysed its genetic diversity and evolutionary dynamics and evaluated viral distribution in tissues of infected animals to identify possible mechanisms of transmission. SKAV was detected in 72.5% (37/51) skunks and was present at high rates at all locations tested across North America. Analysis of the complete genomic sequence of 29 strains showed a clear geographic segregation, frequent recombination and marked differences in the evolutionary dynamics of the major structural (VP2) and non-structural (NS1) proteins. NS1 was characterized by a higher variability and a higher percentage of positively selected codons. This could indicate that antibody-mediated enhancement of infection occurs in SKAV, an infection strategy that may be conserved across amdoparvoviruses. Finally, in situ hybridization revealed virus in epithelium of the gastrointestinal tract, urinary tract and skin, indicating that viral transmission could occur via oronasal, faecal and/or urinary secretions, as well as from skin and hair. The endemicity of SKAV over large geographic distances and its high genetic diversity suggest a long-term virus-host association. Persistent shedding and high environmental stability likely contribute to efficient viral spread, simultaneously offering opportunities for cross-species transmission with consequent risk to sympatric species, including domestic animals and wildlife.

SARS-CoV-2 wildlife surveillance in Ontario and Québec.

Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), the virus responsible for the coronavirus disease 2019 pandemic, is capable of infecting a variety of wildlife species. Wildlife living in close contact with humans are at an increased risk of SARS-CoV-2 exposure and, if infected, have the potential to become a reservoir for the pathogen, making control and management more difficult. The objective of this study is to conduct SARS-CoV-2 surveillance in urban wildlife from Ontario and Québec, increasing our knowledge of the epidemiology of the virus and our chances of detecting spillover from humans into wildlife. Methods: Using a One Health approach, we leveraged activities of existing research, surveillance and rehabilitation programs among multiple agencies to collect samples from 776 animals from 17 different wildlife species between June 2020 and May 2021. Samples from all animals were tested for the presence of SARS-CoV-2 viral ribonucleic acid, and a subset of samples from 219 animals across three species (raccoons, Procyon lotor; striped skunks, Mephitis mephitis; and mink, Neovison vison) were also tested for the presence of neutralizing antibodies. Results: No evidence of SARS-CoV-2 viral ribonucleic acid or neutralizing antibodies was detected in any of the tested samples. Conclusion: Although we were unable to identify positive SARS-CoV-2 cases in wildlife, continued research and surveillance activities are critical to better understand the rapidly changing landscape of susceptible animal species. Collaboration between academic, public and animal health sectors should include experts from relevant fields to build coordinated surveillance and response capacity.

Characterizing the demographic history and prion protein variation to infer susceptibility to chronic wasting disease in a naïve population of white-tailed deer (Odocoileus virginianus).

Assessments of the adaptive potential in natural populations are essential for understanding and predicting responses to environmental stressors like climate change and infectious disease. Species face a range of stressors in human-dominated landscapes, often with contrasting effects. White-tailed deer (Odocoileus virginianus; deer) are expanding in the northern part of their range following decreasing winter severity and increasing forage availability. Chronic wasting disease (CWD), a prion disease affecting deer, is likewise expanding and represents a major threat to deer and other cervids. We obtained tissue samples from free-ranging deer across their native range in Ontario, Canada, which has yet to detect CWD in wild populations. We used high-throughput sequencing to assess neutral genomic variation and variation in the prion protein gene (PRNP) that is partly responsible for the protein misfolding when deer contract CWD. Neutral variation revealed a high number of rare alleles and no population structure, and demographic models suggested a rapid historical population expansion. Allele frequencies of PRNP variants associated with CWD susceptibility and disease progression were evenly distributed across the landscape and consistent with deer populations not infected with CWD. We estimated the selection coefficient of CWD, with simulations showing an observable and rapid shift in PRNP allele frequencies that coincides with the start of a novel CWD outbreak. Sustained surveillance of genomic and PRNP variation can be a useful tool for guiding management practices, which is especially important for CWD-free regions where deer are managed for ecological and economic benefits.

Serologic Evidence of Arthropod-Borne Virus Infections in Wild and Captive Ruminants in Ontario, Canada.

Arthropod-borne viruses (arboviruses) are globally widespread, and their transmission cycles typically involve numerous vertebrate species. Serologic testing of animal hosts can provide a routine surveillance approach to monitoring animal disease systems, can provide a surveillance alternative to arthropod testing and human case reports, and may augment knowledge of epizootiology. Wild and captive ruminants represent good candidate sentinels to track geographic distribution and prevalence of select arboviruses. They often are geographically widespread and abundant, inhabit areas shared by humans and domestic animals, and are readily fed on by various hematophagous arthropod vectors. Ontario, Canada, is home to high densities of coexisting humans, livestock, and wild cervids, as well as growing numbers of arthropod vectors because of the effects of climate change. We collected blood samples from 349 livestock (cattle/sheep) and 217 cervids (wild/farmed/zoo) in Ontario (2016-2019) to assess for antibodies to zoonotic and agriculturally important arboviruses. Livestock sera were tested for antibodies to bluetongue virus (BTV) and epizootic hemorrhagic disease virus (EHDV). Sera from cervids were tested for antibodies to BTV, EHDV, West Nile virus (WNV), eastern equine encephalitis virus (EEEV), Powassan virus (POWV), and heartland virus (HRTV). Fifteen (9.0%) cattle were seropositive for EHDV-serotype 2. Nine (4.2%) cervids were seropositive for arboviruses; three confirmed as WNV, three as EEEV, and one as POWV. All animals were seronegative for BTV and HRTV. These results reveal low seroprevalence of important agricultural, wildlife, and zoonotic pathogens and underline the need for continued surveillance in this and other regions in the face of changing environmental conditions.

A long-distance translocation initiated an outbreak of raccoon rabies in Hamilton, Ontario, Canada.

Despite proactive measures to prevent raccoon rabies entering Canada from the United States, several incursions of this disease have occurred. The largest outbreak, first reported in December 2015 in the city of Hamilton, Ontario, has resulted in the reporting of 449 animal cases as of December 31, 2018. Initial phylogenetic studies on the index case suggested that this outbreak was not due to local cross-border spread from the Niagara region of the United States where raccoon rabies has persisted for several years. Phylogenetic analysis of whole genome sequences of a viral collection from the Hamilton area and several US states indicates that a long-distance translocation of a diseased animal from southeastern New York State was responsible for this incursion. The role of the skunk as a potential secondary host supporting persistence and / or spread of the virus is also examined.

Aleutian mink disease virus in striped skunks (Mephitis mephitis): evidence for cross-species spillover.

Aleutian mink disease virus (AMDV) causes a parvovirus infection, initially characterized in American mink (Neovison vison), that may have harmful effects on wild populations of susceptible animals. In North America, where American mink are native, the origin, host range, and prevalence of AMDV in wild species is not clear. We studied striped skunks (Mephitis mephitis) and raccoons (Procyon lotor) to determine whether species sympatric with mink are potential reservoirs in the transmission of AMDV to wild mink and mink farms. Antibodies to AMDV were detected in 41% of skunk serum samples (143/347) and AMDV nucleic acids were detected in 32% (14/40) of skunk spleen samples by PCR, indicating that AMDV exposure and infection were frequent in skunks. We detected no AMDV antibodies in 144 raccoon blood samples. Phylogenetic analysis revealed a newly identified AMDV haplogroup consisting of isolates from Ontario skunks and a free-ranging domestic mink from Ontario. Our findings of frequent AMDV infection in skunks, close genetic similarity between skunk and mink AMDV isolates, and evidence of AMDV transmission from skunks to mink support the hypothesis that skunks may be acting as alternative hosts and reservoirs of AMDV to wild mink through cross-species virus spillover.

Testing for Aleutian mink disease virus in the river otter (Lontra canadensis) in sympatry with infected American mink (Neovison vison).

Aleutian mink disease virus (AMDV) occurs in the American mink (Neovison vison) in wild populations and on mink farms and can cause illness and death. The North American river otter (Lontra canadensis) may be exposed to AMDV because of shared space and habitat with mink. Using serology and real-time PCR, we tested river otters across Ontario for AMDV infection. We found no evidence of infection in otters, a surprising finding given the sympatric distribution, niche overlap, and close phylogenetic relationship of the river otter and the American mink. Our results are consistent with the hypothesis that the major point of spillover of AMDV between mink farms and wildlife is manure and composting carcasses on mink farms. Mink farms in Ontario are generally in agricultural landscapes; it is unlikely that river otter use these habitats and thus are likely not exposed to AMDV. We found no evidence that AMD is an important disease for the river otters in Ontario.

Physiological adaptations to prolonged fasting in the overwintering striped skunk (Mephitis mephitis).

Wintertime physiology of captive striped skunks (Mephitis mephitis) in response to cold ambient temperature (Ta) and fasting was investigated with body temperature (Tb) and activity recordings and analyses of hematology, plasma biochemistry and tissue fatty acids (FA). After 105 days of food deprivation, the skunks were in phase II of fasting indicated by the elevated plasma nonesterified FA and glycerol but no accumulation of nitrogen end products. Shorter-chain saturated and monounsaturated FA together with C18-20 n-3 polyunsaturated FA were preferentially mobilized. Individual amino acids responded to fasting in a complex manner, while essential and nonessential amino acid sums remained stable. Increases in hemoglobin and hematocrit suggested dehydration. The activity levels were lower in mid-January-early March, and the activity bouts were mostly displayed between 17:00-23:00 h. Daily torpor was observed in two females with 29 and 46 bouts. The deepest torpor (Tb<31 °C) occurred between dawn and early afternoon and lasted for 3.3 ± 0.18 h. The average minimum Tb was 29.2 ± 0.15 °C and the lowest recorded Tb was 25.8 °C. There was significant relation between the average 24-h Tb and Ta. Increases in wintertime Ta, as predicted by climate change scenarios, could influence torpor patterns in the species.

Testing for bias in a sentinel species: contaminants in free-ranging domestic, wild, and hybrid mink.

Sentinel species are important tools for studies of biodiversity and environmental health. The American mink (Neovison vison) has long been considered a sentinel of environmental contamination, since the species is known to be sensitive to a number of common contaminants, including polychlorinated biphenyls (PCB) and mercury. Mink may not always satisfy an important criterion of sentinels however--that they are continuous residents of the environment being sampled. This is because domestic mink commonly escape from farms, and can be confused with wild mink in areas where mink ranching is prevalent, biasing estimates of environmental contamination taken from free-ranging mink samples. We tested for bias in a sample of free-ranging mink from Ontario, Canada, where both genetic ancestry (domestic, wild, and domestic-wild hybrid) and contaminant burdens (PCBs and mercury) were known. Of 133 mink sampled for both contaminants and genetic ancestry, 9% were determined to be domestic and 10.5% hybrid animals. We found that including domestic and hybrid mink in our analysis resulted in overestimating mean PCB burdens in wild mink by 27%, and underestimating mercury by 13%. We also investigated morphological methods to aid in excluding domestic mink from free-ranging mink samples and found that we had the highest classification success using skull size (condylobasal length), which was 15% and 12% greater in male and female domestic than wild mink, respectively. Given the potential use of mink as sentinels, and also the potential for bias, we recommend that researchers take steps to exclude domestic mink from free-ranging mink samples in studies of environmental health.

Molecular epidemiology of Aleutian disease virus in free-ranging domestic, hybrid, and wild mink.

Aleutian mink disease (AMD) is a prominent infectious disease in mink farms. The AMD virus (AMDV) has been well characterized in Europe where American mink (Neovison vison) are an introduced species; however, in North America, where American mink are native and the disease is thought to have originated, the virus' molecular epidemiology is unknown. As such, we characterized viral isolates from Ontario free-ranging mink of domestic, hybrid, and wild origin at two proteins: NS1, a nonstructural protein, and VP2, a capsid protein. AMDV DNA was detected in 25% of free-ranging mink (45 of 183), indicating prevalent active infection. Median-joining networks showed that Ontario AMDV isolates formed two subgroups in the NS1 region and three in the VP2 region, which were somewhat separate from, but closely related to, AMDVs circulating in domestic mink worldwide. Molecular analyses showed evidence of AMDV crossing from domestic to wild mink. Our results suggest that AMDV isolate grouping is linked to both wild endogenous reservoirs and the long-term global trade in domestic mink, and that AMD spills back and forth between domestic and wild mink. As such, biosecurity on mink farms is warranted to prevent transmission of the disease between mink farms and the wild.

Mink farms predict Aleutian disease exposure in wild American mink.

BACKGROUND: Infectious diseases can often be of conservation importance for wildlife. Spillover, when infectious disease is transmitted from a reservoir population to sympatric wildlife, is a particular threat. American mink (Neovison vison) populations across Canada appear to be declining, but factors thus far explored have not fully explained this population trend. Recent research has shown, however, that domestic mink are escaping from mink farms and hybridizing with wild mink. Domestic mink may also be spreading Aleutian disease (AD), a highly pathogenic parvovirus prevalent in mink farms, to wild mink populations. AD could reduce fitness in wild mink by reducing both the productivity of adult females and survivorship of juveniles and adults. METHODS: To assess the seroprevalence and geographic distribution of AD infection in free-ranging mink in relation to the presence of mink farms, we conducted both a large-scale serological survey, across the province of Ontario, and a smaller-scale survey, at the interface between a mink farm and wild mink. CONCLUSIONS/SIGNIFICANCE: Antibodies to AD were detected in 29% of mink (60 of 208 mink sampled); however, seroprevalence was significantly higher in areas closer to mink farms than in areas farther from farms, at both large and small spatial scales. Our results indicate that mink farms act as sources of AD transmission to the wild. As such, it is likely that wild mink across North America may be experiencing increased exposure to AD, via disease transmission from mink farms, which may be affecting wild mink demographics across their range. In light of declining mink populations, high AD seroprevalence within some mink farms, and the large number of mink farms situated across North America, improved biosecurity measures on farms are warranted to prevent continued disease transmission at the interface between mink farms and wild mink populations.