Establishment of a European-type strain of Echinococcus multilocularis in Canadian wildlife.

In 2009, a haplotype closely related to European strains of Echinococcus multilocularis was detected in a dog from the Quesnel region of British Columbia, Canada. We now report the establishment of this same haplotype in 7 coyotes (Canis latrans) trapped within 40 km of Quesnel, BC. In addition, 3 coyotes and 1 red fox (Vulpes vulpes) harboured adult cestodes morphologically compatible with that of E. multilocularis (overall prevalence 33% in 33 carnivores). None of 156 potential intermediate hosts, including 131 representatives of two highly suitable rodent species, Peromyscus maniculatus and Microtus pennsylvanicus, trapped from a region 120-210 km south of Quesnel were infected. This report confirms the establishment and local transmission of a European-type strain of E. multilocularis (the causative agent of human alveolar hydatid disease), in wildlife in a forested region of North America where this cestode had not been previously detected, with significance for public and animal health.

Cryptosporidium and Giardia in marine-foraging river otters (Lontra canadensis) from the Puget Sound Georgia Basin ecosystem.

Species of Cryptosporidium and Giardia can infect humans and wildlife and have the potential to be transmitted between these 2 groups; yet, very little is known about these protozoans in marine wildlife. Feces of river otters (Lontra canadensis), a common marine wildlife species in the Puget Sound Georgia Basin, were examined for species of Cryptosporidium and Giardia to determine their role in the epidemiology of these pathogens. Using ZnSO4 flotation and immunomagnetic separation, followed by direct immunofluorescent antibody detection (IMS/DFA), we identified Cryptosporidium sp. oocysts in 9 fecal samples from 6 locations and Giardia sp. cysts in 11 fecal samples from 7 locations. The putative risk factors of proximate human population and degree of anthropogenic shoreline modification were not associated with the detection of Cryptosporidium or Giardia spp. in river otter feces. Amplification of DNA from the IMS/DFA slide scrapings was successful for 1 sample containing > 500 Cryptosporidium sp. oocysts. Sequences from the Cryptosporidium 18S rRNA and the COWP loci were most similar to the ferret Cryptosporidium sp. genotype. River otters could serve as reservoirs for Cryptosporidium and Giardia species in marine ecosystems. More work is needed to better understand the zoonotic potential of the genotypes they carry as well as their implications for river otter health.

Beyond Rabies: Are Free-Ranging Skunks (Mephitis mephitis) in British Columbia Reservoirs of Emerging Infection?

Wild animal reservoirs are an important source of emerging and zoonotic infection. Skunks (Mephitis mephitis) are a reservoir of skunk strain rabies virus in Canada, with the exception of some areas including the province of British Columbia (BC). Beyond rabies, the reservoir status of skunks for emerging and zoonotic pathogens in BC is unknown. From March 2011 to February 2015, 50 free-ranging skunks were necropsied and tested for 4 pathogens: influenza A, Aleutian disease virus (ADV), Leptospira spp. and Salmonella spp. Two skunks (4%) with respiratory disease caused by influenza A (H1N1) pdm09 were detected during the human flu season suggesting that skunks may represent a target population for reverse zoonosis of this strain of influenza A virus. High prevalence of ADV infection was detected (43/50, 86%). Two of the infected skunks exhibited Aleutian disease (AD) suggesting that skunks act as both a reservoir and a target population for the virus. Most studies of ADV have focused on the potential for infection of free-ranging species living near mink farms. Our study suggests that urban skunks may be a primary host for the virus independent of domestic mink. Whether skunks act as a reservoir of ADV infection for other peridomestic species will depend on host specificity of the viral strains. Leptospira interrogans was detected in 18% (9/49) of the skunks. Identification of the serovar(s) detected is needed to determine any public health risk of leptospirosis following exposure to infected skunks. Salmonella spp. was isolated from three of 43 skunks (7%), specifically S. Typhimurium, S. Muenchen and S. Enteritidis. These serotypes cause disease in humans, but the low prevalence of infection suggests there is a low risk for zoonotic transmission.

Cryptococcus gattii in wildlife of Vancouver Island, British Columbia, Canada.

Although Cryptococcus gattii has emerged as an important pathogen of humans and domestic animals on Vancouver Island, Canada since 1999; its distribution in regional wildlife species is largely unknown. Opportunistic sampling methods were employed to obtain nasal swabs for fungal culture from wild mammal species residing within the coastal Douglas fir biogeoclimatic zone on the southeast coast of the island. Samples were collected from 91 animals representing 14 species. Cryptococcus gattii was isolated from the nasal swabs of two eastern gray squirrels (Sciurus carolinensis) trapped in Duncan, British Columbia. The relative proportion of nasal colonization in wild mammal species is consistent with findings in domestic animals, suggesting that animals may be good indicators of environmental organisms.

Antibodies to selected pathogens in free-ranging terrestrial carnivores and marine mammals in Canada.

Antibody titres to selected pathogens (canine adenovirus [CAV-2], feline herpesvirus [FHV], phocine herpesvirus [PHV-1], canine distemper virus, dolphin morbillivirus [DMV], phocine distemper virus [PDV], parainfluenza virus type 3 [PI3], rabies virus, dolphin rhabdovirus [DRV], canine coronavirus, feline coronavirus, feline leukaemia virus, Borrelia burgdorferi and Toxoplasma gondii) were determined in whole blood or serum samples from selected free-ranging terrestrial carnivores and marine mammals, including cougars (Fellis concolor), lynxes (Fellis lynx), American badgers (Taxidea taxus), fishers (Martes pennanti), wolverines (Gulo gulo), wolves (Canis lupus), black bears (Ursus americanus), grizzly bears (Ursus arctos), polar bears (Ursus maritimus), walruses (Odobenus rosmarus) and belugas (Delphinapterus leucas), which had been collected at several locations in Canada between 1984 and 2001. Antibodies to a number of viruses were detected in species in which these infections have not been reported before, for example, antibodies to CAV-2 in walruses, to PDV in black bears, grizzly bears, polar bears, lynxes and wolves, to DMV in grizzly bears, polar bears, walruses and wolves, to PI3 in black bears and fishers, and to DRV in belugas and walruses.

Molecular study of free-ranging mule deer and white-tailed deer from British Columbia, Canada, for evidence of Anaplasma spp. and Ehrlichia spp.

Twenty-three free-ranging white-tailed deer (WTD; Odocoileus virginianus) and six mule deer (MD; Odocoileus hemionus) from south-central British Columbia, Canada, were tested for Anaplasma marginale by msp5 gene-specific PCR and Ehrlichia spp. by 16S rRNA or citrate synthase (gltA) gene-specific PCR, as well as by PCR with universal 16S rRNA primers detecting a wide range of bacteria. No deer tested positive for A. marginale. Amplification with universal 16S rRNA primers followed by sequencing of cloned fragments detected an Anaplasma sp. in one of 23 (4.3%) WTD and six of six (100%) MD and Bartonella sp. in four of 23 (17.4%) WTD. The Anaplasma sp. was genetically distinct from A. marginale and all other recognized members of the genus. Four of six (66.7%) MD and 0 of 23 (0%) WTD were Ehrlichia positive by PCR with primers for 16S rRNA and gltA genes. The sequences of gltA PCR fragments were identical to each other and to the respective region of the gltA gene of an Ehrlichia sp. which we detected previously in naturally infected cattle from the same area, suggesting the possibility of biological transmission of this rickettsia between cattle and wild cervids. Antibodies reactive with the MSP5 protein of A. marginale were detected using a competitive enzyme-linked immunosorbent assay in two of six (33.3%) MD, but not in WTD. The two seropositive MD were PCR positive for both the Anaplasma sp. and Ehrlichia sp. detected in this study, suggesting a reaction of antibodies against one or both of these rickettsias with the MSP5 antigen.

Population genetic structure of North American thinhorn sheep (Ovis dalli).

The thinhorn sheep (Ovis dalli ssp.) provides a rare example of a North American large mammal that occupies most of its native range and maintains close to ancestral population size. There are currently two recognized subspecies, Dall's sheep (O. d. dalli) and Stone's sheep (O. d. stonei), the validity of which remains uncertain. We investigated the spatial genetic structure of thinhorn sheep populations representing both subspecies by genotyping individuals (n = 919) from across the species range at 12 variable microsatellite loci. We found high levels of genetic diversity within (HE = 0.722) and significant genetic structure among the 24 sampled areas (FST = 0.160). Genetic distance measures and Bayesian clustering analyses revealed the presence of at least eight subpopulations that are delineated by mountain range topology. A strong overall pattern of isolation-by-distance is evident across the sampling range (r = 0.75, P < 0.001) suggesting limited dispersal and extensive philopatry. Partial Mantel tests of this relationship showed mountain range distinctions represent significant barriers to gene flow (P = 0.0001), supporting the Bayesian analyses. Genetic structure was more strongly pronounced in southern Yukon and Alaska than elsewhere. We also show evidence for genetic differences between the two currently recognized thinhorn subspecies.

Potential contamination of drinking water with Toxoplasma gondii oocysts.

The world's first documented toxoplasmosis outbreak associated with a municipal water supply was recognized in 1995 in Victoria, British Columbia, Canada. It was hypothesized that domestic cat (Felis catus) or cougar (Felis concolor) faeces contaminated a surface water reservoir with Toxoplasma gondii oocysts. An extensive investigation of the Victoria watershed 1 year following the outbreak documented the presence of an endemic T. gondii cycle involving the animals inhabiting the area. Cats and cougars were observed throughout the watershed. Serological evidence of T. gondii infection was demonstrated among domestic cats living in the Victoria area. Cougars were found to shed T. gondii oocysts. Serological evidence of T. gondii infection in deer mice living in the riparian environments of the watershed suggested that T. gondii oocysts were being shed near the water edge. Contamination of Victoria's water supply with T. gondii oocysts potentially occurred during the study period and future waterborne toxoplasmosis outbreaks in this and other communities are possible.