Association of Environmental Factors with Seasonal Intensity of Erysipelothrix rhusiopathiae Seropositivity among Arctic Caribou.

Several caribou (Rangifer tarandus) populations have been declining concurrently with increases in infectious diseases in the Arctic. Erysipelothrix rhusiopathiae, a zoonotic bacterium, was first described in 2015 as a notable cause of illness and death among several Arctic wildlife species. We investigated epidemiologic and environmental factors associated with the seroprevalence of E. rhusiopathiae in the Arctic and found that seropositivity was highest during warmer months, peaking in September, and was highest among adult males. Summer seroprevalence increases tracked with the oestrid index from the previous year, icing and snowing events, and precipitation from the same year but decreased with growing degree days in the same year. Seroprevalence of E. rhusiopathiae varied more during the later years of the study. Our findings provide key insights into the influence of environmental factors on disease prevalence that can be instrumental for anticipating and mitigating diseases associated with climate change among Arctic wildlife and human populations.

A PRELIMINARY ASSESSMENT OF THE EFFECTS OF REPRODUCTIVE SEASONALITY ON SEMINAL PLASMA CHARACTERISTICS IN NORTH AMERICAN BISON (BISON BISON).

The two subspecies of the North American bison, plains ( Bison bison bison) and wood ( Bison bison athabascae) bison, are seasonal breeders. The objective of this study was to conduct a preliminary investigation into the effects of season on semen. To test the hypothesis that there are seasonal effects on seminal plasma, protein profiles of seminal plasma from plains and wood bison (n = 2 of each subspecies) were compared between breeding and nonbreeding seasons. Using two-dimensional gel electrophoresis of seminal plasma proteins, 54 of 170 spots (plains bison) and 19 of 153 spots (wood bison) had differential expression (≥2-fold; P < 0.01) between seasons. Based on immunoblotting, BSP5 and TIMP-2 (two fertility-associated proteins in cattle) were higher during the breeding vs. nonbreeding season. Furthermore, epididymal sperm incubated with seminal plasma from the nonbreeding season had lower postthaw progressive motility (17.33 ± 7.47 vs. 22.09 ± 6.67%; mean ± SD) and an increased ability to undergo a lysophosphatidylcholine-induced acrosome reaction (77.83 ± 8.47 vs. 52.67 ± 7.76%; mean ± SD) as compared to epididymal sperm incubated with seminal plasma from the breeding season. In a heterologous in vitro fertilization system (using bovine oocytes), cleavage rate was higher for sperm exposed to seminal plasma from the breeding vs. the nonbreeding season (75.35 ± 16.55 vs. 33.63 ± 12.44%; mean ± SD). This study suggested that differential expression of seminal plasma characteristics modulating sperm function is one of the mechanisms by which reproductive seasonality affects sperm function in the North American bison.

Echinococcus multilocularis and Echinococcus canadensis in wolves from western Canada.

Echinococcus species are important parasites of wildlife, domestic animals and people worldwide; however, little is known about the prevalence, intensity and genetic diversity of Echinococcus tapeworms in Canadian wildlife. Echinococcus tapeworms were harvested from the intestines of 42% of 93 wolves (Canis lupus) from five sampling regions in the Northwest Territories, Manitoba and Saskatchewan, and visually identified to genus level by microscopic examination. Genetic characterization was successful for tapeworms from 30 wolves, and identified both Echinococcus canadensis and Echinococcus multilocularis in all sampling locations. Mixed infections of E. canadensis/E. multilocularis, as well as the G8/G10 genotypes of E. canadensis were observed. These findings suggest that wolves may be an important definitive host for both parasite species in western Canada. This represents the first report of wolves naturally infected with E. multilocularis in North America, and of wolves harbouring mixed infections with multiple species and genotypes of Echinococcus. These observations provide important information regarding the distribution and diversity of zoonotic species of Echinococcus in western North America, and may be of interest from public health and wildlife conservation perspectives.

Direct and indirect costs of parasitism preceding a population decline of an Arctic ungulate.

Parasites negatively affect the fitness of ungulate hosts directly, and in wild ungulates, these effects may be synzootic with other stressors, such as limited nutritional resources. In the Arctic, muskoxen (Ovibos moschatus) occur in a highly seasonal environment and must rely on finite energetic resources for survival and productivity. We investigated the costs of gastrointestinal nematodes on the body condition and reproductive status of 141 muskoxen, on Banks Island, Canada, when the population was at a peak in numbers and density. Using a Partial Least Squares Path Modelling approach, we found that high adult nematode abundance was associated with lower body condition, and high parasite abundance was associated with female reproduction including the indirect effect through on body condition (n = 87). These findings suggest that individuals prioritize energetic reserves for reproduction over parasite defence. In fall 2003, a severe icing event that restricted access to forage was associated with high overwinter mortality of muskoxen and a population crash. Through direct and indirect costs of parasite infection on body condition and reproduction, the high abundance of parasites may have contributed to the effects of this extreme weather event. Understanding the mechanisms in which parasites impact fitness can help explain the ecological drivers of ungulate populations and predict the interactions between the environment and populations.

Contrasting results of culture-dependent and molecular analyses of Mycobacterium avium subsp. paratuberculosis from wood bison.

Reduced to near extinction in the late 1800s, a number of wood bison populations (Bison bison athabascae) have been re-established through reintroduction initiatives. Although an invaluable tool for conservation, translocation of animals can spread infectious agents to new areas or expose animals to pathogens in their new environment. Mycobacterium avium subsp. paratuberculosis, a bacterium that causes chronic enteritis in ruminants, is among the pathogens of potential concern for wood bison management and conservation. In order to inform translocation decisions, our objectives were to determine the M. avium subsp. paratuberculosis infection status of wood bison herds in Canada and to culture and genetically characterize the infective strain(s). We tested fecal samples from bison (n = 267) in nine herds using direct PCR for three M. avium subsp. paratuberculosis-specific genetic targets with different copy numbers within the M. avium subsp. paratuberculosis genome. Restriction enzyme analysis (REA) and sequencing of IS1311 were performed on seven samples from five different herds. We also evaluated a panel of different culture conditions for their ability to support M. avium subsp. paratuberculosis growth from feces and tissues of direct-PCR-positive animals. Eighty-one fecal samples (30%) tested positive using direct IS900 PCR, with positive samples from all nine herds; of these, 75% and 21% were also positive using ISMAP02 and F57, respectively. None of the culture conditions supported the growth of M. avium subsp. paratuberculosis from PCR-positive samples. IS1311 REA and sequencing indicate that at least two different M. avium subsp. paratuberculosis strain types exist in Canadian wood bison. The presence of different M. avium subsp. paratuberculosis strains among wood bison herds should be considered in the planning of translocations.

An improved method for the extraction and quantification of adult Echinococcus from wildlife definitive hosts.

The scraping and counting technique (SCT), with sensitivity values close to 100 %, has been the protocol recommended by global regulatory bodies for the extraction of Echinococcus cestodes from the intestines of wild carnivores. The proposed scraping, filtration and counting technique (SFCT) maintained the sensitivity (p = 0.801, α = 0.05) and increased the efficiency of sample processing. SCT had sensitivity and negative predictive value of 91 and 97 %, respectively, when compared to SFCT. The SFCT significantly decreased processing time (p = 0.0001, α = 0.05) for each sample. The SFCT took an average of 68.5 min less to quantify than SCT, as the SFCT samples consistently contained less debris. The SFCT is therefore appealing for general post-mortem surveillance, to determine if prevalence and intensity of infection are changing in an established region, or if these important parasitic zoonoses are newly established in a region or host species.

An outbreak of bovine tuberculosis in an intensively managed conservation herd of wild bison in the Northwest Territories.

An outbreak of bovine tuberculosis was detected in the Hook Lake Wood Bison Recovery Project captive-breeding herd in March 2005. This study investigates the most likely source of Mycobacterium bovis and identifies difficulties associated with salvaging tuberculosis-free animals from an endemically infected herd.

Evaluation and delivery of domestic animal health services in remote communities in the Northwest Territories: A case study of status and needs.

Domestic animal health services are supplied to communities in Canada's Northwest Territories (NT) in diverse ways, including private veterinary practices in 2 of 33 communities, and by mail-order, fly-in, free clinics, and a government-coordinated lay vaccinator program in some of the other 31 communities. We evaluated delivery, needs, and potential uptake of domestic animal health services in the Sahtu Settlement Area, NT by offering free clinics for 225 dogs in 2008 and 2009; and administered questionnaires to 42 dog owners and 67 students in 2008. Owners indicated that 20% of dogs were neutered, 37% had had rabies vaccinations, and 29% had been dewormed. Physical examination of dogs demonstrated that 54% were "thin" and 4% were "emaciated." Owners and youth showed a range of attitudes toward dogs and supported improved domestic animal health services. Future services need to build on existing programs and collaborate with communities to ensure relevance, ownership, and sustainability.

Novel insights into serodiagnosis and epidemiology of Erysipelothrix rhusiopathiae, a newly recognized pathogen in muskoxen (Ovibos moschatus).

BACKGROUND: Muskoxen are a key species of Arctic ecosystems and are important for food security and socio-economic well-being of many Indigenous communities in the Arctic and Subarctic. Between 2009 and 2014, the bacterium Erysipelothrix rhusiopathiae was isolated for the first time in this species in association with multiple mortality events in Canada and Alaska, raising questions regarding the spatiotemporal occurrence of the pathogen and its potential impact on muskox populations. MATERIALS AND METHODS: We adapted a commercial porcine E. rhusiopathiae enzyme-linked immunosorbent assay to test 958 blood samples that were collected from muskoxen from seven regions in Alaska and the Canadian Arctic between 1976 and 2017. The cut-off between negative and positive results was established using mixture-distribution analysis, a data-driven approach. Based on 818 samples for which a serological status could be determined and with complete information, we calculated trends in sample seroprevalences in population time-series and compared them with population trends in the investigated regions. RESULTS: Overall, 219/818 (27.8%, 95% Confidence Interval: 24.7-31.0) samples were classified as positive for exposure to E. rhusiopathiae. There were large variations between years and regions. Seropositive animals were found among the earliest serum samples tested; 1976 in Alaska and 1991 in Canada. In Alaskan muskoxen, sample seroprevalence increased after 2000 and, in two regions, peak seroprevalences occurred simultaneously with population declines. In one of these regions, concurrent unusual mortalities were observed and E. rhusiopathiae was isolated from muskox carcasses. In Canada, there was an increase in sample seroprevalence in two muskox populations following known mortality events that had been attributed to E. rhusiopathiae. CONCLUSION: Our results indicate widespread exposure of muskoxen to E. rhusiopathiae in western Canada and Alaska. Although not new to the Arctic, we documented an increased exposure to the pathogen in several regions concurrent with population declines. Understanding causes for the apparent increased occurrence of this pathogen and its association with large scale mortality events for muskoxen is critical to evaluate the implications for wildlife and wildlife-dependent human populations in the Arctic.

Correction: Novel insights into serodiagnosis and epidemiology of Erysipelothrix rhusiopathiae, a newly recognized pathogen in muskoxen (Ovibos moschatus).

[This corrects the article DOI: 10.1371/journal.pone.0231724.].

Hiding in plain sight: discovery and phylogeography of a cryptic species of Trichinella (Nematoda: Trichinellidae) in wolverine (Gulo gulo).

Understanding parasite diversity and distribution is essential in managing the potential impact of parasitic diseases in animals and people. Imperfect diagnostic methods, however, may conceal cryptic species. Here, we report the discovery and phylogeography of a previously unrecognized species of Trichinella in wolverine (Gulo gulo) from northwestern Canada that was indistinguishable from T. nativa using the standard multiplex PCR assay based on the expansion segment 5 (ESV) of ribosomal DNA. The novel genotype, designated as T13, was discovered when sequencing the mitochondrial genome. Phylogenetic analyses of the mitochondrial genome and of 15 concatenated single copy orthologs of nuclear DNA indicated a common ancestor for the encapsulated clade is shared by a subclade containing Trichinella spiralis and Trichinella nelsoni, and a subclade containing T13 and remaining taxa: T12 + (T2 + T6) + [(T5 + T9) + (T3 + T8)]. Of 95 individual hosts from 12 species of mammalian carnivores from northwestern Canada from which larvae were identified as T. nativa on multiplex PCR, only wolverines were infected with T13 (14 of 42 individuals). These infections were single or mixed with T. nativa and/or T6. Visual examination and motility testing confirmed that T13 is encapsulated and likely freeze-tolerant. We developed a new Polymerase Chain Reaction-Restriction Fragment Length Polymorphism which unequivocally distinguishes between T13 and T. nativa. We propose Trichinella chanchalensis n. sp. for T13, based on significant genetic divergence from other species of Trichinella and broad-based sampling of the Trichinella genome. Exploration of Alaskan and Siberian isolates may contribute to further resolution of a phylogeographically complex history for species of Trichinella across Beringia, including Trichinella chanchalensis n. sp. (T13).

Risk factors and prevalence of antibodies for Toxoplasma gondii in diaphragmatic fluid in wolverines (Gulo gulo) from the Northwest Territories, Canada.

Toxoplasma gondii, a zoonotic food borne parasite that can infect almost all warm-blooded animals including people, and ranks 4th among 24 most significant global foodborne parasites listed by the World Health Organization/United Nations Food and Agriculture Organization (FAO/WHO, 2014). Exposure to T. gondii has been reported in wildlife and people in the Canadian North, despite low densities of feline definitive hosts. The ecology of this host-parasite system could be affected by changing climate and landscape in boreal and sub-Arctic regions, and surveillance data are critically needed. Wolverines are an economically and culturally important species in northern Canada due to their valuable fur. Fluid obtained from diaphragmatic muscle of 127 wolverines (Gulo gulo) were tested for antibodies to T. gondii using an enzyme linked immunosorbent assay (ELISA). A seroprevalence of 62% (Confidence Interval (CI): 53-71%) was observed. This result indicates high levels of exposure, likely either through environmental contamination with T. gondii oocysts shed by infected wild felids, or consumption of carcasses/offal of other intermediate hosts containing tissue cysts with bradyzoites in tissues. We examined factors associated with seropositivity, including age, sex, harvest location, harvest location with respect to treeline, and body condition index. Adult (≥2 years) wolverines had 5.2 times higher odds of being sero-positive than juvenile (<1 years) wolverines. The highest seroprevalence was observed in wolverines from Sahtu and South Slave regions. Proportion of sero-positive wolverines harvested above and below the tree line was not significantly different (60% vs 65%). Age was the only significant predictor of T. gondii exposure in wolverines (using logistic regression analysis); further studies should target larger sample sizes. This study is an example of how fluid from diaphragmatic muscle can be used for screening for T. gondii antibodies in wolverines. The diaphragm, commonly collected for screening for another food borne parasite, Trichinella, in wildlife harvested for human consumption, can be used for screening of T. gondii exposure in wildlife. Due to their predatory and scavenging lifestyle and high trophic level, wolverines could serve as a sentinel species for T. gondii.

A Transdisciplinary Approach to Brucella in Muskoxen of the Western Canadian Arctic 1989-2016.

Brucella serostatus was evaluated in 3189 muskoxen sampled between 1989 and 2016 from various locations of the Canadian Arctic archipelago and mainland, near the communities of Sachs Harbour and Ulukhaktok, Northwest Territories, and Cambridge Bay and Kugluktuk, Nunavut. Brucella antibodies were found only in muskoxen sampled around Cambridge Bay, both on southern Victoria Island and on the adjacent mainland (Kent Peninsula). Consistent with participatory epidemiology data documented from local harvesters describing increased Brucella-like syndromes (swollen joints and lameness) and a decreased proportion of juveniles, the apparent Brucella seroprevalence in the sampled muskoxen of the Cambridge Bay area increased from 0.9% (95% CI 0.3-2.1) in the period of 1989-2001 to 5.6% (95% CI 3.3-8.9) in 2010-2016. The zoonotic bacteria Brucella suis biovar 4 was also cultured from tissues of muskoxen sampled on Victoria Island near Ulukhaktok in 1996 (n = 1) and Cambridge Bay in 1998, 2014, and 2016 (n = 3). Overall, our data demonstrate that B. suis biovar 4 is found in muskoxen that are harvested for food and by guided hunts on Victoria Island and Kent Peninsula, adding an important public health dimension to this study. Robust participatory epidemiology data on muskox health and diseases greatly enhanced the interpretation of our Cambridge Bay data and, combined with the serological and microbiological data, provide compelling evidence that the prevalence of B. suis biovar 4 has increased in this area since the late 1990s. This study enhances the available knowledge on Brucella exposure and infection in muskoxen and provides an example of how scientific knowledge and local knowledge can work together to better understand disease status in wildlife.

Trichinella pseudospiralis in a wolverine (Gulo gulo) from the Canadian North.

Species of Trichinella are a globally distributed assemblage of nematodes, often with distinct host ranges, which include people, domestic, and wild animals. Trichinella spp. are important in northern Canada, where dietary habits of people and methods of meat preparation (drying, smoking, fermenting as well as raw) increase the risk posed by these foodborne zoonotic parasites. Outbreaks in the arctic and subarctic regions of Canada and the United States are generally attributed to T. nativa (T2) or the T6 genotype, when genetic characterization is performed. We report the discovery of Trichinella pseudospiralis (T4), a non-encapsulated species, in a wolverine (Gulo gulo) from the Northwest Territories of Canada. This parasite has been previously reported elsewhere from both mammals and carnivorous birds, but our findings represent new host and geographic records for T. pseudospiralis. Multiplex PCR and sequencing of fragments of Cytochrome Oxidase Subunit I (COI) and D3 rDNA confirmed the identification. Phylogenetically, Canadian isolates linked with each other and others derived from Palearctic or Neotropical regions, but not elsewhere in the Nearctic (continental USA). We suggest that migratory birds might have played a role in the dispersal of this pathogen 1000's of km to northwestern Canada. Wolverines are not typically consumed by humans, and thus should not pose a direct food safety risk for trichinellosis. However, the current finding suggests that they may serve as an indicator of a broader distribution for T. pseudospiralis. Along with infection risk already recognized for T. nativa and Trichinella T6, our observations emphasize the need for further studies using molecular diagnostics and alternative methods to clarify if this is a solitary case, or if T. pseudospiralis and other freeze susceptible species of Trichinella (such as T. spiralis) circulate more broadly in wildlife in Canada, and elsewhere.

The global distribution of Bacillus anthracis and associated anthrax risk to humans, livestock and wildlife.

Bacillus anthracis is a spore-forming, Gram-positive bacterium responsible for anthrax, an acute infection that most significantly affects grazing livestock and wild ungulates, but also poses a threat to human health. The geographic extent of B. anthracis is poorly understood, despite multi-decade research on anthrax epizootic and epidemic dynamics; many countries have limited or inadequate surveillance systems, even within known endemic regions. Here, we compile a global occurrence dataset of human, livestock and wildlife anthrax outbreaks. With these records, we use boosted regression trees to produce a map of the global distribution of B. anthracis as a proxy for anthrax risk. We estimate that 1.83 billion people (95% credible interval (CI): 0.59-4.16 billion) live within regions of anthrax risk, but most of that population faces little occupational exposure. More informatively, a global total of 63.8 million poor livestock keepers (95% CI: 17.5-168.6 million) and 1.1 billion livestock (95% CI: 0.4-2.3 billion) live within vulnerable regions. Human and livestock vulnerability are both concentrated in rural rainfed systems throughout arid and temperate land across Eurasia, Africa and North America. We conclude by mapping where anthrax risk could disrupt sensitive conservation efforts for wild ungulates that coincide with anthrax-prone landscapes.

Mycobacterium avium subspecies paratuberculosis in Bison (Bison bison) from Northern Canada.

Nested polymerase chain reaction (PCR) using the Mycobacterium avium subspecies paratuberculosis (Map)-specific region, locus 251, was used as a screening tool for the detection of Map DNA in fecal samples from northern Canadian bison herds. Further characterization of positive samples (26/835) was performed because Map DNA was found without signs of disease. Strain typing, using PCR-Restriction endonucleas assay (REA), was limited to two samples but revealed that the samples corresponded to a cattle-related strain and a sheep-related strain. Sequencing of part of the IS1311 region from the two samples revealed a unique three base-pair region, which is only found within the northern Canadian bison isolates.

Radiocesium in caribou and reindeer in northern Canada, Alaska and Greenland from 1958 to 2000.

This study summarizes the spatial and temporal trends of fallout (137)Cs concentrations in caribou and reindeer (Rangifer tarandus ssp.), reported in various programs in Canada, Alaska and Greenland, over a 40-y period. During the 1960s, the highest (137)Cs concentrations (2000-3000Bqkg(-1) wet weight in muscle) were found in the large caribou herds of central northern Canada, with levels about 50% lower in Alaska and Greenland. Concentrations varied by up to a factor of 6 between spring and fall. Concentrations in reindeer were comparable to those in caribou from the same regions. The highest (137)Cs concentrations (750Bqkg(-1)) in the late 1980s were found in the Caniapiscau herd of central Québec. The contribution from the Chernobyl accident in 1986 was estimated from (134)Cs measurements to be 10-40% of the total (137)Cs. Present concentrations have declined to 200-300Bqkg(-1). The effective half-life (T(eff)) of (137)Cs in Canadian caribou herds was estimated to be about 6 y. The potential impact on the health of northern residents is discussed.

ANTHRAX IN THE MACKENZIE WOOD BISON (BISON BISON ATHABASCAE) POPULATION: 2012 ANTHRAX OUTBREAK AND HISTORICAL EXPOSURE IN NONOUTBREAK YEARS.

Anthrax, caused by the spore-forming bacterium Bacillus anthracis, poses a threat to wood bison (Bison bison athabascae) conservation. We used descriptive epidemiology to characterize a large outbreak of anthrax in the Mackenzie bison population in the Northwest Territories, Canada, in 2012 and investigated historical serologic exposure of the bison to the bacterium in nonoutbreak years. Between late June and early August 2012, 451 bison carcasses were detected; mortality peaked from 13-19 July. A substantial number of calves, yearlings, and adult females died in the 2012 outbreak, unlike in two previous anthrax outbreaks in this population that killed mostly mature males. On the basis of the difference in estimates of population size prior to the outbreak (2012) and after the outbreak (2013), it is possible that not all dead bison were found during the outbreak. We assessed serologic history of exposure to B. anthracis by using samples from the Mackenzie wood bison population collected between 1986 and 2009. Overall, 87 of 278 samples were positive (31%). Seroprevalence was lower in females (18%, 10/55) than males (36%, 72/203). The highest proportion of positive submissions (90%) was from 1994, the year following the only anthrax outbreak within the historical data set. Both adult males and females had a higher likelihood of being seropositive than the younger age categories. There was a trend toward declining antibody levels between the 1993 and 2012 outbreak years.

Trichinella Surveillance in Black Bears ( Ursus americanus ) from the Dehcho Region, Northwest Territories, Canada, 2002-15.

We used muscle digestion to test black bears ( Ursus americanus ) from the southwestern Northwest Territories, Canada, for Trichinella. Results showed a prevalence of 4.1%. Some bears had infection intensities of more than one larva per gram of muscle tissue; this level in meat is considered to pose a human consumption safety risk.