Recent Hybridization between a Polar Bear and Grizzly Bears in the Canadian Arctic

Grizzly bears have recently become more common on the Arctic Islands in the Inuvialuit Settlement Region, concurrently with a period of environmental change. Over the last decade, grizzly bear – polar bear hybrids have been confirmed within this region, triggering extensive discussion and speculation regarding the impact of hybridization on the parent species. Through harvests, sightings, and captures, we document an increase in the presence of grizzly bears and combine field observations of hybrids with genetic analysis and parentage analysis to identify four first-generation (F1) hybrids and four offspring of F1 hybrids and grizzly bears (backcross-to-grizzly-bear individuals). We trace these eight hybrid individuals to a single female polar bear who mated with two grizzly bears. We sampled one of her mates on the sea ice in the High Arctic and deduced the genotype of the other from his five offspring. The two male grizzly bears are sires of both the F1 generation and the backcross-to-grizzly-bear generation. So what initially appeared to be a sudden spate of hybridization in the western Canadian Arctic originated with the unusual mating between three non-hybrid parents. The breakdown of species barriers may start with atypical mating preferences of select individuals; however, the story we present can be traced to a single female polar bear who, along with three of her known F1 offspring, has been killed.La présence du grizzli se fait plus courante dans l’archipel Arctique de la région désignée des Inuvialuit depuis un certain temps, ce qui coïncide avec une période de changement environnemental. Ces dix dernières années, la présence d’ours polaires-grizzlis hybrides a été confirmée dans cette région, ce qui a déclenché d’importantes discussions et hypothèses relativement aux incidences de l’hybridation sur les espèces apparentées. Au moyen de récoltes, d’observations et de captures, nous avons documenté l’augmentation de la présence de grizzlis, alliées à des observations d’hybrides sur le terrain avec analyse génétique et analyse de parenté afin d’identifier quatre hybrides de première génération (F1) et quatre descendants d’hybrides F1 et de grizzlis (individus issus de rétrocroisements avec un grizzli). Nous faisons remonter ces huit hybrides à une seule ourse polaire qui s’est accouplée avec deux grizzlis. Nous avons prélevé un échantillon d’un de ses compagnons sur la glace de mer de l’Extrême-Arctique et avons déduit le génotype de l’autre à partir de ses cinq descendants. Les deux grizzlis mâles sont des géniteurs de la génération F1 et de la génération du rétrocroisement avec un grizzli. Donc, ce qui semblait être, au début, une montée soudaine d’hybridation dans l’ouest de l’Arctique canadien est attribuable à l’accouplement inhabituel de trois parents non hybrides. La rupture des obstacles auxquels les espèces font face pourrait commencer par les préférences d’accouplement atypiques de certains individus. Toutefois, la situation dont il est ici question remonte à une seule ourse polaire qui a été tuée, avec trois de ses descendants connus de la génération F1

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. pseu- dospiralis. 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 mi- gratory birds might have played a role in the dispersal of this pathogen 1000\u27s of km to northwestern Canada. Wolverines are not typically consumed by humans, and thus should not pose a direct food safety risk for tri- chinellosis. 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 observa- tions 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

Supporting Information: Influence of Carbon and Lipid Sources on Variation of Mercury and Other Trace Elements in Polar Bears (\u3ci\u3eUrsus maritimus\u3c/i\u3e)

MATERIALS AND METHODS Stable Carbon Isotope Analysis and Quality Control Fatty Acids Analysis and Quality Control Mercury and Other Trace Metal and Element Analysis and Quality Control REFERENCE SECTIO

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 para- sitic 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 stan- dard 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 com- mon 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) + (T 3 + 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 phylogeo- graphically complex history for species of Trichinella across Beringia, including Trichinella chanchalensis n. sp. (T13

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. pseu- dospiralis. 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 mi- gratory birds might have played a role in the dispersal of this pathogen 1000\u27s of km to northwestern Canada. Wolverines are not typically consumed by humans, and thus should not pose a direct food safety risk for tri- chinellosis. 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 observa- tions 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

Recent Hybridization between a Polar Bear and Grizzly Bears in the Canadian Arctic

Grizzly bears have recently become more common on the Arctic Islands in the Inuvialuit Settlement Region, concurrently with a period of environmental change. Over the last decade, grizzly bear – polar bear hybrids have been confirmed within this region, triggering extensive discussion and speculation regarding the impact of hybridization on the parent species. Through harvests, sightings, and captures, we document an increase in the presence of grizzly bears and combine field observations of hybrids with genetic analysis and parentage analysis to identify four first-generation (F1) hybrids and four offspring of F1 hybrids and grizzly bears (backcross-to-grizzly-bear individuals). We trace these eight hybrid individuals to a single female polar bear who mated with two grizzly bears. We sampled one of her mates on the sea ice in the High Arctic and deduced the genotype of the other from his five offspring. The two male grizzly bears are sires of both the F1 generation and the backcross-to-grizzly-bear generation. So what initially appeared to be a sudden spate of hybridization in the western Canadian Arctic originated with the unusual mating between three non-hybrid parents. The breakdown of species barriers may start with atypical mating preferences of select individuals; however, the story we present can be traced to a single female polar bear who, along with three of her known F1 offspring, has been killed.La présence du grizzli se fait plus courante dans l’archipel Arctique de la région désignée des Inuvialuit depuis un certain temps, ce qui coïncide avec une période de changement environnemental. Ces dix dernières années, la présence d’ours polaires-grizzlis hybrides a été confirmée dans cette région, ce qui a déclenché d’importantes discussions et hypothèses relativement aux incidences de l’hybridation sur les espèces apparentées. Au moyen de récoltes, d’observations et de captures, nous avons documenté l’augmentation de la présence de grizzlis, alliées à des observations d’hybrides sur le terrain avec analyse génétique et analyse de parenté afin d’identifier quatre hybrides de première génération (F1) et quatre descendants d’hybrides F1 et de grizzlis (individus issus de rétrocroisements avec un grizzli). Nous faisons remonter ces huit hybrides à une seule ourse polaire qui s’est accouplée avec deux grizzlis. Nous avons prélevé un échantillon d’un de ses compagnons sur la glace de mer de l’Extrême-Arctique et avons déduit le génotype de l’autre à partir de ses cinq descendants. Les deux grizzlis mâles sont des géniteurs de la génération F1 et de la génération du rétrocroisement avec un grizzli. Donc, ce qui semblait être, au début, une montée soudaine d’hybridation dans l’ouest de l’Arctique canadien est attribuable à l’accouplement inhabituel de trois parents non hybrides. La rupture des obstacles auxquels les espèces font face pourrait commencer par les préférences d’accouplement atypiques de certains individus. Toutefois, la situation dont il est ici question remonte à une seule ourse polaire qui a été tuée, avec trois de ses descendants connus de la génération F1

\u3ci\u3eProtostrongylus stilesi\u3c/i\u3e (Nematoda: Protostrongylidae): Ecological Isolation and Putative Host-Switching Between Dall’s Sheep and Muskoxen in a Contact Zone

The occurrence of Protostrongylus stilesi in a population of introduced muskoxen, Ovibos moschatus wardi, on the Arctic Coastal Plain, Yukon Territory (YT) and Northwest Territories (NT), Canada, is consistent with a contemporary colonization event from Dall’s sheep, Ovis dalli dalli, which indicates that host specificity may be ecologically based and contextual for this parasite. Colonization of muskoxen by P. stilesi may be a predictable event in zones of sympatry with Dall’s sheep; exposure to infection may coincide with occupation of winter ranges of Dall’s sheep by muskoxen during the summer season. Disruption of contemporary ecological isolating barriers can result from anthropogenically or climatologically driven habitat perturbation, and result from management practices that influence the distribution of ungulate hosts. Thus, if zones of contact become more extensive or the temporal limits on allopatry are relaxed, we may observe increasing instances of host switching by parasites or pathogens at the interface of newly emerging ecotones. Impacts to northern systems linked to climatologically and anthropogenically driven global change and the effects of management must be tracked within the context of biodiversity survey and inventory and archival collections, as foundations for monitoring ecosystem-level perturbations. A developing interface for muskoxen, wild sheep, and parasites along the Mackenzie River ecotone represents a natural model or field laboratory to examine these processes. Additonally, lungworms, Protostrongylus spp., had not been reported in muskoxen, and a new geographic record for this nematode was established in Dall’s sheep from the northern Richardson Mountains, NT

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 para- sitic 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 stan- dard 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 com- mon 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) + (T 3 + 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 phylogeo- graphically complex history for species of Trichinella across Beringia, including Trichinella chanchalensis n. sp. (T13

A new multiplexed magnetic capture—Droplet digital PCR tool for monitoring wildlife population health and pathogen surveillance

Abstract Anthropogenic stressors are exacerbating the emergence and spread of pathogens worldwide. In regions like the Arctic, where ecosystems are particularly susceptible, marked changes are predicted in regional diversity, intensity, and patterns of infectious diseases. To understand such rapidly changing host‐pathogen dynamics and mitigate the impacts of novel pathogens, we need sensitive disease surveillance tools. We developed and validated a novel multiplexed, magnetic capture, and ddPCR tool for the surveillance of multiple pathogens in polar bears, a sentinel species that is considered susceptible to climate change and other stressors with a pan‐Arctic distribution. Through sequence‐specific magnetic capture, we concentrated five target template sequences from three zoonotic bacteria (Erysipelothrix rhusiopathiae, Francisella tularensis, and Mycobacterium tuberculosis complex) and two parasitic (Toxoplasma gondii and Trichinella spp.) pathogens from large quantities (<100 g) of host tissue. We then designed and validated two multiplexed probe‐based ddPCR assays for the amplification and detection of the low‐concentration target DNA. Validations used 48 polar bear tissues (muscle and liver). We detected 14, 1, 3, 4, and 22 tissue positives for E. rhusiopathiae, F. tularensis, M. tuberculosis complex, T. gondii, and Trichinella spp., respectively. These multiplexed assays offer a rapid, specific tool for quantifying and monitoring the changing geographical and host distributions of pathogens relevant to human and animal health