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BACKGROUND: Canine leptospirosis is a zoonotic disease of global importance. The identification of social and environmental risk factors remains limited and is needed for planning interventions. Our objectives were to summarise the global trends in social and environmental risk factors for canine leptospirosis and to identify knowledge gaps. METHODS: We searched peer-reviewed and grey literature for observational studies on canine leptospirosis written in English. Variables assessed via formal statistical tests were recorded and categorised into thematic risk factor groups. The risk factors were summarised by the number of unique studies and their direction of association. RESULTS: Of the 3184 articles identified, 66 met the inclusion criteria. Eight environmental and three social risk factor groups were identified. Exposure to animals and water was commonly studied, but social risk factors were infrequently examined. Most studies were conducted on populations in higher-income countries and countries in the Americas. LIMITATIONS: Several limitations were encountered, including inconsistent study designs and measurement and control of variables as well as disproportionate representation of studies from the Americas and higher-income countries. CONCLUSION: The weight and quality of evidence for social and environmental risk factors were highly variable. Additional research following standard reporting guidelines to improve transparency and increase generalisability to different contexts is needed.

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One Health is an integrated approach that aims to balance and optimize the interconnectedness of the health of humans, animals, and ecosystems. Using this transdisciplinary approach, experts from across Canada led the formation of the Wildlife Emerging Pathogens Initiative (Wild EPI) to undertake research and surveillance programs evaluating the potential risks of emerging pathogens at the human-animal interface. Wild EPI is dedicated to implementing the One Health approach to enhance our understanding of the epidemiology and burden of zoonotic infections among humans and other animal hosts.

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The objectives of this research were to assess ingested plastics and accumulated heavy metals in four urban gull species. Additionally, the relationships between ingested plastics and selected demographic and health metrics were assessed. Between 2020-2021 during the non-breeding seasons, 105 gulls (46 American herring gulls (HERG, Larus argentatus smithsonianus), 39 great black-backed gulls (GBBG, Larus marinus), 16 Iceland gulls (Larus glaucoides), 4 glaucous gulls (Larus hyperboreus)) were killed at a landfill in coastal Newfoundland and Labrador, Canada, as part of separate, permitted kill-to-scare operations related to aircraft safety. Birds were necropsied, the upper gastrointestinal tract contents were processed using standard techniques, and livers were analyzed for accumulated As, Cd, Hg, and Pb. The relationships between ingested plastics, demographics, and health metrics were assessed in HERG and GBBG. Across all four species, 85 % of birds had ingested at least one piece of anthropogenic debris, with 79 % ingesting at least one piece of plastic. We detected interspecific differences in plastic ingestion and hepatic trace metals, with increased ingested plastics detected in GBBG compared with HERG. For GBBG, levels of ingested plastic were relatively greater for birds with higher scaled mass index, while HERG with more ingested plastic had higher liver lead concentrations.

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Following the detection of novel highly pathogenic avian influenza virus (HPAIV) H5N1 clade 2.3.4.4b in Newfoundland, Canada, in late 2021, avian influenza virus (AIV) surveillance in wild birds was scaled up across Canada. Herein, we present the results of Canada's Interagency Surveillance Program for Avian Influenza in Wild Birds during the first year (November 2021-November 2022) following the incursions of HPAIV from Eurasia. The key objectives of the surveillance program were to (i) identify the presence, distribution, and spread of HPAIV and other AIVs; (ii) identify wild bird morbidity and mortality associated with HPAIV; (iii) identify the range of wild bird species infected by HPAIV; and (iv) genetically characterize detected AIV. A total of 6,246 sick and dead wild birds were tested, of which 27.4% were HPAIV positive across 12 taxonomic orders and 80 species. Geographically, HPAIV detections occurred in all Canadian provinces and territories, with the highest numbers in the Atlantic and Central Flyways. Temporally, peak detections differed across flyways, though the national peak occurred in April 2022. In an additional 11,295 asymptomatic harvested or live-captured wild birds, 5.2% were HPAIV positive across 3 taxonomic orders and 19 species. Whole-genome sequencing identified HPAIV of Eurasian origin as most prevalent in the Atlantic Flyway, along with multiple reassortants of mixed Eurasian and North American origins distributed across Canada, with moderate structuring at the flyway scale. Wild birds were victims and reservoirs of HPAIV H5N1 2.3.4.4b, underscoring the importance of surveillance encompassing samples from sick and dead, as well as live and harvested birds, to provide insights into the dynamics and potential impacts of the HPAIV H5N1 outbreak. This dramatic shift in the presence and distribution of HPAIV in wild birds in Canada highlights a need for sustained investment in wild bird surveillance and collaboration across interagency partners. IMPORTANCE: We present the results of Canada's Interagency Surveillance Program for Avian Influenza in Wild Birds in the year following the first detection of highly pathogenic avian influenza virus (HPAIV) H5N1 on the continent. The surveillance program tested over 17,000 wild birds, both sick and apparently healthy, which revealed spatiotemporal and taxonomic patterns in HPAIV prevalence and mortality across Canada. The significant shift in the presence and distribution of HPAIV in Canada's wild birds underscores the need for sustained investment in wild bird surveillance and collaboration across One Health partners.

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[This corrects the article DOI: 10.1371/journal.pone.0260234.].

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OBJECTIVE: Bartonella are emerging bacterial zoonotic pathogens. Utilization of clotted blood samples for surveillance of these bacteria in wildlife has begun to supersede the use of tissues; however, the efficacy of these samples has not been fully investigated. Our objective was to compare the efficacy of spleen and blood samples for DNA extraction and direct detection of Bartonella spp. via qPCR. In addition, we present a protocol for improved DNA extraction from clotted, pelleted (i.e., centrifuged) blood samples obtained from wild small mammals. RESULTS: DNA concentrations from kit-extracted blood clot samples were low and A260/A280 absorbance ratios indicated high impurity. Kit-based DNA extraction of spleen samples was efficient and produced ample DNA concentrations of good quality. We developed an in-house extraction method for the blood clots which resulted in apposite DNA quality when compared to spleen samples extracted via MagMAX DNA Ultra 2.0 kit. We detected Bartonella in 9/30 (30.0%) kit-extracted spleen DNA samples and 11/30 (36.7%) in-house-extracted blood clot samples using PCR. Our results suggest that kit-based methods may be less suitable for DNA extraction from blood clots, and that blood clot samples may be superior to tissues for Bartonella detection.

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Wildlife disease surveillance, particularly for pathogens with zoonotic potential such as Highly Pathogenic Avian Influenza Virus (HPAIV), is critical to facilitate situational awareness, inform risk, and guide communication and response efforts within a One Health framework. This study evaluates the intensity of avian influenza virus (AIV) surveillance in Ontario's wild bird population following the 2021 H5N1 incursion into Canada. Analyzing 2562 samples collected between November 1, 2021, and October 31, 2022, in Ontario, Canada, we identify spatial variations in surveillance intensity relative to human population density, poultry facility density, and wild mallard abundance. Using the spatial scan statistic, we pinpoint areas where public engagement, collaborations with Indigenous and non-Indigenous hunter/harvesters, and working with poultry producers, could augment Ontario's AIV wild bird surveillance program. Enhanced surveillance at these human-domestic animal-wildlife interfaces is a crucial element of a One Health approach to AIV surveillance. Ongoing assessment of our wild bird surveillance programs is essential for strategic planning and will allow us to refine approaches and generate results that continue to support the program's overarching objective of safeguarding the health of people, animals, and ecosystems.

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Plastic ingestion presents many potential avenues of risk for wildlife. Understanding which species and environments are most exposed to plastic pollution is a critical first step in investigating the One Health implications of plastic exposure. The objectives of this study were the following: 1) Utilize necropsy as part of ongoing passive disease surveillance to investigate ingested mesoplastics in birds collected in Ontario and Nunavut, and examine the relationships between bird-level factors and ingested debris; 2) evaluate microplastic ingestion compared to ingested mesodebris in raptors; and 3) identify potential sentinel species for plastic pollution monitoring in understudied freshwater and terrestrial (inland) environments. Between 2020 and 2022, 457 free-ranging birds across 52 species were received for postmortem examination. The upper gastrointestinal tracts were examined for mesoplastics and other debris (>2 mm) using standard techniques. Twenty-four individuals (5.3%) retained mesodebris and prevalence varied across species, with foraging technique, food type, and foraging substrate all associated with different metrics of debris ingestion. The odds of ingesting any type of anthropogenic mesodebris was nine times higher for non-raptorial species than for raptors. For a subset of raptors (N = 54) across 14 species, the terminal portion of the distal intestinal tract was digested with potassium hydroxide and microparticles were assessed using stereo-microscopy. Although only one of 54 (1.9%) raptors included in both analyses retained mesodebris in the upper gastrointestinal tract, 24 (44.4%) contained microparticles in the distal intestine. This study demonstrates that a variety of Canadian bird species ingest anthropogenic debris in inland systems, and suggests that life history and behaviour are associated with ingestion risk. For raptors, the mechanisms governing exposure and ingestion of mesoplastics appear to be different than those that govern microplastics. Herring gulls (Larus argentatus) and ring-billed gulls (Larus delawarensis) are proposed as ideal sentinels for plastic pollution monitoring in inland systems.

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Echinococcus multilocularis, a cestode with zoonotic potential, is now known to have a high prevalence in wild canid definitive hosts of southern Ontario. The distribution of E. multilocularis across this region in red foxes (Vulpes vulpes) and coyotes (Canis latrans) is widespread yet heterogenous. In contrast, confirmed diagnoses of E. multilocularis in wild free-ranging intermediate hosts within Ontario are currently limited to a single eastern chipmunk (Tamias striatus). These findings prompted ongoing surveillance efforts in intermediate host species, primarily rodents. Our report describes the results of passive surveillance through wildlife carcass submissions to the Canadian Wildlife Health Cooperative (CWHC) and targeted active sampling of small mammal species from 2018 to 2023; a second and third eastern chipmunk were found to be infected with E. multilocularis. However, these were the only occurrences from surveillance efforts which collectively totaled 510 rodents and other small mammals. Continued surveillance for E. multilocularis in intermediate hosts is of high importance in light of the recent emergence of this parasite in Ontario.

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AIMS: Rat-associated zoonotic pathogen transmission at the human-wildlife interface is a public health concern in urban environments where Norway rats (Rattus norvegicus) thrive on abundant anthropogenic resources and live in close contact with humans and other animal species. To identify potential factors influencing zoonotic pathogen occurrence in rats, we investigated associations between environmental and sociodemographic factors and Leptospira interrogans and Bartonella spp. infections in rats from Windsor, Ontario, Canada, while controlling for the potential confounding effects of animal characteristics (i.e., sexual maturity and body condition). METHODS AND RESULTS: Between November 2018 and June 2021, 252 rats were submitted by collaborating pest control professionals. Kidney and spleen samples were collected for L. interrogans and Bartonella spp. PCR and sequencing, respectively. Of the rats tested by PCR, 12.7% (32/252) were positive for L. interrogans and 16.3% (37/227) were positive for Bartonella species. Associations between infection status and environmental and sociodemographic variables of interest were assessed via mixed multivariable logistic regression models with a random intercept for social group and fixed effects to control for sexual maturity and body condition in each model. The odds of L. interrogans infection were significantly higher in rats from areas with high building density (odds ratio [OR]: 3.76; 95% CI: 1.31-10.79; p = 0.014), high human population density (OR: 3.31; 95% CI: 1.20-9.11; p = 0.021), high proportion of buildings built in 1960 or before (OR: 11.21; 95% CI: 2.06-60.89; p = 0.005), and a moderate number of reports of uncollected garbage compared to a low number of reports (OR: 4.88; 95% CI: 1.01-23.63; p = 0.049). A negative association was observed between median household income and Bartonella spp. infection in rats (OR: 0.26; 95% CI: 0.08-0.89; p = 0.031). CONCLUSIONS: Due to the complexity of the ecology of rat-associated zoonoses, consideration of environmental and sociodemographic factors is of critical importance to better understand the nuances of host-pathogen systems and inform how urban rat surveillance and intervention efforts should be distributed within cities.

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Aquatic bird bornavirus 1 (ABBV-1) has a high prevalence of infection in certain North American populations of Canada geese (Branta canadensis), suggesting a possible role of these birds as an ABBV-1 reservoir. The goal of this study was to evaluate the ability of Canada geese to become experimentally infected with ABBV-1, develop lesions, and transmit the virus to conspecifics. One-week-old Canada geese (n, 65) were inoculated with ABBV-1 through the intramuscular (IM) or cloacal (CL) routes, with the control group receiving carrier only. An additional 6 geese were added to each group to test horizontal transmission (sentinel birds). Geese were monitored daily, and selected birds were euthanized at 1, 8, and 15-weeks post infection (wpi) to assess virus replication in tissues and lesion development. At 15 wpi, over 70% of IM birds were infected, while the CL route yielded only 1 infected goose. Of the infected IM geese, 26% developed encephalitis and/or myelitis after 8 wpi. No clinical signs were observed, and no sentinel birds became infected in any group. Only 1 oropharyngeal swab (IM group) tested positive for ABBV-1 RNA, while the water from the enclosures was consistently negative for virus RNA. This study documents successful experimental infection of Canada geese with ABBV-1, with findings comparable to what is described in infection trials with other waterfowl species. However, minimal shedding and lack of environmental dispersal indicate that Canada geese have little potential to disseminate the virus among wild waterfowl, and that other species could be better suited to act as chronic ABBV-1 shedders in the wild.

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Aquatic bird bornavirus 1 (ABBV-1) is a neurotropic virus that causes persistent infection in the nervous system of wild waterfowl. This study evaluated whether Pekin ducks, the most common waterfowl raised worldwide, are susceptible to ABBV-1 infection and associated disease. Groups of Pekin ducks were inoculated with ABBV-1 through the intracranial (IC; n, 32), intramuscular (IM; n, 30), and choanal (CH; n, 30) routes. Controls (CO; n, 29) received carrier only. At 1, 12, and 21 weeks postinfection (wpi), 7-14 birds were euthanized to assess virus distribution and lesions. Infection rates in the IC and IM groups were over 70%, while only 4 ducks in the CH group became infected. Neurological signs were observed in 8 ducks only, while over 25% of IC and IM birds had encephalitis and/or myelitis. Seroconversion was highest in the IC and IM groups, and mucosal ABBV-1 RNA shedding was most frequent in the IC group (53%). None of the fertile eggs laid during the experiment tested positive for ABBV-1 RNA. This study shows that Pekin ducks are permissive to ABBV-1 infection and partly susceptible to associated disease. While mucosal shedding may be an important route of transmission, congenital infection appears unlikely.

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We tested liver samples from 372 Norway rats (Rattus norvegicus) from southern Ontario, Canada, during 2018-2021 to investigate presence of hepatitis E virus infection. Overall, 21 (5.6%) rats tested positive for the virus. Sequence analysis demonstrated all infections to be rat hepatitis E virus (Rocahepevirus ratti genotype C1).

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Increasing reports of canine distemper virus (CDV) in a variety of hosts, and changing CDV dynamics, have led to renewed interest in the ecology of CDV infections in wildlife. Longitudinal serologic studies provide insights into intrapopulation and intraindividual pathogen dynamics, but few studies in wildlife have been conducted. We used data from 235 raccoons (Procyon lotor) captured on more than one occasion between May 2011 and November 2013 to investigate CDV dynamics in Ontario, Canada. Using mixed multivariable logistic regression, we found that juvenile raccoons were more likely to be seronegative from August to November than from May to July. Using paired titers from CDV-exposed individual raccoons, we determined that the winter breeding season, when there is high intraspecific contact and an increase in susceptible juveniles, may be a period of high risk for CDV exposure. Interestingly, CDV seropositive adult raccoons had nondetectable titers ranging from 1 mo to 1 yr later. Based on our preliminary investigation using two different statistical approaches, CDV exposure was associated with a decrease in parvovirus titer. This result raises important questions about whether virus-induced immune amnesia occurs after CDV exposure, which has been described for measles virus, a closely related pathogen. Overall, our results provide significant insights into CDV dynamics. Further research is needed to investigate whether CDV-induced immune amnesia occurs in raccoons and to determine the potential impacts of a reduced population immunity that may occur secondary to CDV exposure, particularly as it relates to rabies control efforts.

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BACKGROUND: Culicoides Latreille (Diptera: Ceratopogonidae) is a genus of hematophagous midges feeding on various vertebrate hosts and serving as a vector for numerous pathogens important to livestock and wildlife health. North American pathogens include bluetongue (BT) and epizootic hemorrhagic disease (EHD) viruses. Little is known about Culicoides spp. distribution and abundance and species composition in Ontario, Canada, despite bordering numerous U.S. states with documented Culicoides spp. and BT and EHD virus activity. We sought to characterize Culicoides spp. distribution and abundance and to investigate whether select meteorological and ecological risk factors influenced the abundance of Culicoides biguttatus, C. stellifer, and the subgenus Avaritia trapped throughout southern Ontario. METHODS: From June to October of 2017 to 2018, CDC-type LED light suction traps were placed on twelve livestock-associated sites across southern Ontario. Culicoides spp. collected were morphologically identified to the species level when possible. Associations were examined using negative binomial regression among C. biguttatus, C. stellifer, and subgenus Avaritia abundance, and select factors: ambient temperature, rainfall, primary livestock species, latitude, and habitat type. RESULTS: In total, 33,905 Culicoides spp. midges were collected, encompassing 14 species from seven subgenera and one species group. Culicoides sonorensis was collected from three sites during both years. Within Ontario, the northern trapping locations had a pattern of seasonal peak abundance in August (2017) and July (2018), and the southern locations had abundance peaks in June for both years. Culicoides biguttatus, C. stellifer, and subgenus Avaritia were significantly more abundant if ovine was the primary livestock species at trapping sites (compared to bovine). Culicoides stellifer and subgenus Avaritia were significantly more abundant at mid- to high-temperature ranges on trap days (i.e., 17.3-20.2 and 20.3-31.0 °C compared to 9.5-17.2 °C). Additionally, subgenus Avaritia were significantly more abundant if rainfall 4 weeks prior was between 2.7 and 20.1 mm compared to 0.0 mm and if rainfall 8 weeks prior was between 0.1 and 2.1 mm compared to 0.0 mm. CONCLUSIONS: Results from our study describe Culicoides spp. distribution in southern Ontario, the potential for spread and maintenance of EHD and BT viruses, and concurrent health risks to livestock and wildlife in southern Ontario in reference to certain meteorological and ecological risk factors. We identified that Culicoides spp. are diverse in this province, and appear to be distinctly distributed spatially and temporally. The livestock species present, temperature, and rainfall appear to have an impact on the abundance of C. biguttatus, C. stellifer, and subgenus Avaritia trapped. These findings could help inform targeted surveillance, control measures, and the development of management guides for Culicoides spp. and EHD and BT viruses in southern Ontario, Canada.

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Since first being introduced for public use in the 1960s, plastic has become one of the most pervasive and ubiquitous forms of pollution globally. The potential fate and effects of plastic pollution on birds is a rapidly growing area of research, but knowledge of terrestrial and freshwater species is limited. Birds of prey have been particularly understudied, with no published data on plastic ingestion in raptors in Canada to date, and very few studies globally. To assess the ingestion of plastic in raptors, we analysed the contents of the upper gastrointestinal tracts from a total of 234 individuals across 15 raptor species, collected between 2013 and 2021. Upper gastrointestinal tracts were assessed for plastics and anthropogenic particles > 2 mm in size. Of the 234 specimens examined, only five individuals across two species had evidence of retained anthropogenic particles in the upper gastrointestinal tract. Two of 33 bald eagles (Haliaeetus leucocephalus, 6.1%) had retained plastics in the gizzard, while three of 108 barred owls (Strix varia, 2.8%) had retained plastic and non-plastic anthropogenic litter. The remaining 13 species were negative for particles > 2 mm in size (N = 1-25). These results suggest that most hunting raptor species do not appear to ingest and retain larger anthropogenic particles, though foraging guild and habitat may influence risk. We recommend that future research investigate microplastic accumulation in raptors, in order to gain a more holistic understanding of plastic ingestion in these species. Future work should also focus on increasing sample sizes across all species to improve the ability to assess landscape- and species-level factors that influence vulnerability and susceptibility of plastic pollution ingestion.

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Information concerning risk factors associated with Giardia infection in dogs in southern Ontario, Canada, is currently lacking. This study therefore aimed to identify risk factors for Giardia infection in dogs that visit off-leash dog parks in southern Ontario. From May-November 2018, fecal samples were collected from 466 dogs in 12 off-leash dog parks in the Niagara and Hamilton regions of Ontario. A survey that asked questions pertaining to travel history (i.e., area of residence, locations and regions visited in the previous 6 months), basic medical history (i.e., spay/neuter status, veterinary visits, use of deworming medication), consumption of a raw diet, and the physical (i.e., age, sex, breed) and behavioral characteristics (i.e., off-leash activities, hunting activities) of each dog sampled was administered to the respective owner. All fecal samples were examined with the Giardia plate ELISA (IDEXX Laboratories) for parasite antigen. Multivariable logistic regression analyses were conducted on the survey data to investigate putative risk factors for Giardia infection. Overall, 11.8% (95% CI: 9.2-15.1%) of samples tested positive for Giardia antigen. Results from the multivariable logistic regression analyses identified an interaction between dog age and spay/neuter status that was significantly associated with Giardia infection. The odds of infection were greater in intact as compared to neutered adult dogs (OR: 3.6, 95% CI: 1.7-7.9, p = 0.001), and in neutered juvenile dogs as compared to neutered adults (OR: 5.2, 95% CI: 2.2-12.2, p < 0.001). The results provide veterinarians with evidence-based information for identifying dogs at greatest risk of Giardia infection in southern Ontario.

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Lyme disease risk areas have increased across Canada in recent decades with the ongoing range expansion of Ixodes scapularis and Borrelia burgdorferi. Different methodologies are used by federal and provincial governments to determine local Lyme disease risk, which can make comparisons between regions challenging. In this study, seven Canadian Lyme disease risk classification methodologies were compared with each other to highlight the strengths and limitations of how each definition measured I. scapularis and B. burgdorferi risk. Each methodology was applied to active surveillance data from Ontario, and per cent agreement and kappa statistics were calculated. The methodologies varied in their measurements of the risk of exposure to I. scapularis and B. burgdorferi based on their use of active surveillance techniques, multiple types of collected surveillance data and laboratory confirmation of B. burgdorferi. Most initial Lyme disease risk site classifications were maintained over time. Kappa and per cent agreement statistics highlighted large differences between 8 of the 15 methodology pairings, indicating the presence of inconsistencies between most methodologies. Accurate, consistent surveillance and assessment of the spread of I. scapularis and its pathogens will aid with communicating Lyme disease risk to the public and preventing tick-borne pathogen transmission.

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The health and welfare of wild animals are of increasing concern, yet there are very few large-scale data syntheses examining how causes of wildlife morbidity and mortality vary across time, space, and taxa. Records for 18,540 animals submitted to the Canadian Wildlife Health Cooperative (CWHC) (2009-19) and 144,846 animals admitted to 19 wildlife rehabilitation centers (WRCs) (2015-19) were evaluated to 1) identify the main causes of morbidity and mortality for Canadian wildlife and 2) assess the utility and complementarity of these two data sources to further our understanding of wildlife health. The CWHC cases (mortality) were examined by pathologists and grouped by the presence or absence of five diagnostic categories: trauma, emaciation, infection or inflammation, toxicity, and other. These CWHC animals were also classified as "killed due to real or perceived human-wildlife conflict" based on finder history. The WRC admissions were categorized by health issue (according to intake records) and based on reported or observed situational reasons for admission: parental loss, unsafe or unsuitable location, nest or habitat disturbance, illegal possession, and abnormal behavior. For both datasets, the main reason for submission or admission was trauma (44 and 48%, respectively), especially vehicle collisions (7 and 11%) and window or building strikes (5 and 7%). Many other WRC admissions were due to parental loss (28%), cat attacks (6%), and immature animals being found in unsafe or unsuitable locations (6%). Most other CWHC mortalities were caused by infections (27%) and emaciation (23%). Relatively few birds, amphibians, and reptiles submitted to CWHC were killed due to human-wildlife conflict, but 22% of mammals were killed for this reason, highlighting the taxonomic differences in the perceived threat of wildlife to finders, and therefore their response. Together, these data sources highlight key issues impacting the health and welfare of wild animals in Canada.

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