REVENGE OF THE TREES: ENVIRONMENTAL DETERMINANTS AND POPULATION EFFECTS OF INFECTIOUS DISEASE OUTBREAKS ON A BREEDING COLONY OF DOUBLE-CRESTED CORMORANTS (PHALACROCORAX AURITUS) OVER A PERIOD OF 21 YEARS.

During 16 of 21 consecutive annual breeding seasons, two diseases, Newcastle disease and avian cholera, killed approximately 50% of juvenile Double-crested Cormorants (Phalacrocorax auritus) in a large nesting colony in Canada. From 1994 to 2014, we recorded data annually on disease occurrence, causal pathogens, species and age classes affected, total number of breeding pairs of cormorants on the colony site, and other biological parameters. A mathematical model of pathogen transmission was constructed to assess the potential importance of transmission parameters and to test a hypothesis regarding the potential effect of the observed progressive loss of nest trees and the consequent shift from tree-nesting to ground-nesting behavior. The model indicated that juveniles from ground nests were 14 times more likely to die from epidemic disease (50.14% mortality) than were juveniles from nests in trees (3.57% mortality). Additive disease-related mortality of juvenile cormorants in the observed range of 40-60% would reduce a closed cormorant population over time. There was no directional change in the colony population during the study period, suggesting that immigration had compensated for disease-related mortality. Our results highlight the preeminent influence of environmental factors on pathogen transmission and the value of long-term data sets.

Pathology of wild Norway rats in Vancouver, Canada.

To achieve a contemporary understanding of the common and rare lesions that affect wild, urban Norway rats ( Rattus norvegicus), we conducted a detailed pathology analysis of 672 rats from Vancouver, British Columbia, Canada. Grossly evident lesions, such as wounds, abscesses, and neoplasms, were present in 71 of 672 rats (11%) and tended to be severe. The most common and significant lesions were infectious and inflammatory, most often affecting the respiratory tract and associated with bite wounds. We assessed a subset of rats (up to n = 406 per tissue) for the presence of microscopic lesions in a variety of organ systems. The most frequent lesions that could impact individual rat health included cardiomyopathy (128 of 406; 32%), chronic respiratory tract infections as indicated by pulmonary inducible bronchus-associated lymphoid tissue (270 of 403; 67%), tracheitis (192 of 372; 52%), and thyroid follicular hyperplasia (142 of 279; 51%). We isolated 21 bacterial species from purulent lesions in rats with bacterial infections, the most frequent of which were Escherichia coli, Enterococcus sp., and Staphylococcus aureus. Parasitic diseases in rats resulted from infection with several invasive nematodes: Capillaria hepatica in the liver (242 of 672; 36%), Eucoleus sp. in the upper gastrointestinal tract (164 of 399; 41%), and Trichosomoides crassicauda in the urinary bladder (59 of 194; 30%). Neoplastic, congenital, and degenerative lesions were rare, which likely reflects their adverse effect on survival in the urban environment. Our results establish a baseline of expected lesions in wild urban rats, which may have implications for urban rat and zoonotic pathogen ecology, as well as rat control in cities worldwide.

The ecology of avian influenza viruses in wild dabbling ducks (Anas spp.) in Canada.

Avian influenza virus (AIV) occurrence and transmission remain important wildlife and human health issues in much of the world, including in North America. Through Canada's Inter-Agency Wild Bird Influenza Survey, close to 20,000 apparently healthy, wild dabbling ducks (of seven species) were tested for AIV between 2005 and 2011. We used these data to identify and evaluate ecological and demographic correlates of infection with low pathogenic AIVs in wild dabbling ducks (Anas spp.) across Canada. Generalized linear mixed effects model analyses revealed that risk of AIV infection was higher in hatch-year birds compared to adults, and was positively associated with a high proportion of hatch-year birds in the population. Males were more likely to be infected than females in British Columbia and in Eastern Provinces of Canada, but more complex relationships among age and sex cohorts were found in the Prairie Provinces. A species effect was apparent in Eastern Canada and British Columbia, where teal (A. discors and/or A. carolinensis) were less likely to be infected than mallards (A. platyrhynchos). Risk of AIV infection increased with the density of the breeding population, in both Eastern Canada and the Prairie Provinces, and lower temperatures preceding sampling were associated with a higher probability of AIV infection in Eastern Canada. Our results provide new insights into the ecological and demographic factors associated with AIV infection in waterfowl.

ECOLOGICAL DETERMINANTS OF AVIAN INFLUENZA VIRUS, WEST NILE VIRUS, AND AVIAN PARAMYXOVIRUS INFECTION AND ANTIBODY STATUS IN BLUE-WINGED TEAL (ANAS DISCORS) IN THE CANADIAN PRAIRIES.

The Canadian prairies are one of the most important breeding and staging areas for migratory waterfowl in North America. Hundreds of thousands of waterfowl of numerous species from multiple flyways converge in and disperse from this region annually; therefore this region may be a key area for potential intra- and interspecific spread of infectious pathogens among migratory waterfowl in the Americas. Using Blue-winged Teal (Anas discors, BWTE), which have the most extensive migratory range among waterfowl species, we investigated ecologic risk factors for infection and antibody status to avian influenza virus (AIV), West Nile virus (WNV), and avian paramyxovirus-1 (APMV-1) in the three prairie provinces (Alberta, Saskatchewan, and Manitoba) prior to fall migration. We used generalized linear models to examine infection or evidence of exposure in relation to host (age, sex, body condition, exposure to other infections), spatiotemporal (year, province), population-level (local population densities of BWTE, total waterfowl densities), and environmental (local pond densities) factors. The probability of AIV infection in BWTE was associated with host factors (e.g., age and antibody status), population-level factors (e.g., local BWTE population density), and year. An interaction between age and AIV antibody status showed that hatch year birds with antibodies to AIV were more likely to be infected, suggesting an antibody response to an active infection. Infection with AIV was positively associated with local BWTE density, supporting the hypothesis of density-dependent transmission. The presence of antibodies to WNV and APMV-1 was positively associated with age and varied among years. Furthermore, the probability of being WNV antibody positive was positively associated with pond density rather than host population density, likely because ponds provide suitable breeding habitat for mosquitoes, the primary vectors for transmission. Our findings highlight the importance of spatiotemporal, environmental, and host factors at the individual and population levels, all of which may influence dynamics of these and other viruses in wild waterfowl populations.

Demographic and Spatiotemporal Patterns of Avian Influenza Infection at the Continental Scale, and in Relation to Annual Life Cycle of a Migratory Host.

Since the spread of highly pathogenic avian influenza (HPAI) H5N1 in the eastern hemisphere, numerous surveillance programs and studies have been undertaken to detect the occurrence, distribution, or spread of avian influenza viruses (AIV) in wild bird populations worldwide. To identify demographic determinants and spatiotemporal patterns of AIV infection in long distance migratory waterfowl in North America, we fitted generalized linear models with binominal distribution to analyze results from 13,574 blue-winged teal (Anas discors, BWTE) sampled in 2007 to 2010 year round during AIV surveillance programs in Canada and the United States. Our analyses revealed that during late summer staging (July-August) and fall migration (September-October), hatch year (HY) birds were more likely to be infected than after hatch year (AHY) birds, however there was no difference between age categories for the remainder of the year (winter, spring migration, and breeding period), likely due to maturing immune systems and newly acquired immunity of HY birds. Probability of infection increased non-linearly with latitude, and was highest in late summer prior to fall migration when densities of birds and the proportion of susceptible HY birds in the population are highest. Birds in the Central and Mississippi flyways were more likely to be infected compared to those in the Atlantic flyway. Seasonal cycles and spatial variation of AIV infection were largely driven by the dynamics of AIV infection in HY birds, which had more prominent cycles and spatial variation in infection compared to AHY birds. Our results demonstrate demographic as well as seasonal, latitudinal and flyway trends across Canada and the US, while illustrating the importance of migratory host life cycle and age in driving cyclical patterns of prevalence.

Lesions associated with Eucoleus sp. in the non-glandular stomach of wild urban rats (Rattus norvegicus).

Histological lesions associated with Eucoleus sp. infection of the non-glandular stomach were discovered in a wild, urban population of Norway rats (Rattus norvegicus) trapped over a 1-year period in Vancouver, Canada. Four distinct categories of histological lesions in the non-glandular stomach were identified in association with infection in a sample of 183 rats. The apparent prevalence of Eucoleus sp. in the upper gastrointestinal tract (ventral tongue, oropharynx, esophagus and non-glandular stomach) was 43.1% (79/183). Infection with Eucoleus sp. was significantly associated with hyperkeratosis, mucosal hyperplasia, keratin pustules and submucosal inflammation in the non-glandular stomach (P < 0.05). Eucoleus sp. infection and/or related stomach pathology was present in 135/183 (73.8%) of rats. Statistical analysis showed the odds of being affected by Eucoleus sp. or associated stomach pathology were greater in heavier (OR = 1.06, 95% CI = 1.00-1.12) and sexually mature rats (OR = 4.64, 95% CI = 1.23-17.10). Eucoleus sp. infection is common in wild rats in Vancouver and induces substantial host response. The impact of Eucoleus sp. and associated lesions on the health of individual rats and the population as a whole remains to be investigated.

Capillaria hepatica in wild Norway rats (Rattus norvegicus) from Vancouver, Canada.

Capillaria hepatica is a parasitic nematode that infects the liver of rats (Rattus spp.), and occasionally other mammalian species, including humans. Despite its broad geographic distribution and host range, the ecology of this parasite remains poorly understood. We characterized the ecology of C. hepatica in urban Norway rats (Rattus norvegicus) in Vancouver, Canada. The overall prevalence of C. hepatica among Norway rats was 36% (241/671); however, there was significant variation in prevalence among city blocks. Using a generalized linear mixed model to control for clustering by block (where OR is odds ratio and CI is confidence interval), we found C. hepatica infection was negatively associated with season (spring [OR=0.14, 95% CI=0.05-0.39]; summer [OR=0.14, 95% CI=0.03-0.61]; winter [OR=0.34, 95% CI=0.13-0.84], compared to fall) and positively associated with sexual maturity (OR: 7.29, 95% CI=3.98-13.36) and presence of cutaneous bite wounds (OR=1.87, 95% CI=1.11-3.16). Our understanding of the ecology of C. hepatica in rats is hindered by a paucity of data regarding the main mechanisms of transmission (e.g., environmental exposure vs. active cannibalism). However, associations among infection, season, maturity, and bite wounds could suggest that social interactions, possibly including cannibalism, may be important in transmission.

Bait trapping linked to higher avian influenza virus detection in wild ducks.

In 2007, we assessed whether trapping method influenced apparent prevalence of low pathogenic avian influenza viruses (AIV) in wild ducks sampled during Canada's Inter-agency Wild Bird Influenza Survey. Combined cloacal and oropharyngeal swabs were collected from 514 ducks captured by bait trapping (356) and netting from airboats (158), and tested by real-time reverse transcriptase polymerase chain reaction for influenza type A viruses. When controlling for species and capture site, ducks caught in bait traps were 2.6 times more likely to test positive for AIV compared with those netted from airboats (95% CI=1.2-6.0). If bait trapping increases AIV transmission among artificially aggregated ducks, this could have important implications for interpretation of disease surveillance results and waterfowl management programs.

Detection of low pathogenic avian influenza viruses in wild ducks from Canada: comparison of two sampling methods.

Surveillance for avian influenza viruses in wild birds was initiated in Canada in 2005. In 2006, in order to maximize detection of highly pathogenic avian influenza viruses, the sampling protocol used in Canada's Inter-agency Wild Bird Influenza Survey was changed. Instead of collecting a single cloacal swab, as previously done in 2005, cloacal and oropharyngeal swabs were combined in a single vial at collection. In order to compare the two sampling methods, duplicate samples were collected from 798 wild dabbling ducks (tribe Anatini) in Canada between 24 July and 7 September 2006. Low pathogenic avian influenza viruses were detected significantly more often (P<0.0001) in combined oropharyngeal and cloacal samples (261/798, 33%) than in cloacal swabs alone (205/798, 26%). Compared to traditional single cloacal samples, combined samples improved virus detection at minimal additional cost.

PrioNet Canada: a network of centres of excellence for research on prion diseases--ongoing and future research directions.

It is PrioNet's vision to build a network that shapes and sustains prion research in Canada, translating basic science into accessible socioeconomic benefits for global betterment. PrioNet's research is developing surveillance measures, diagnostic tools, vaccines, and potential therapies and determining the various impacts of prion diseases on people. PrioNet seeks to integrate scientifically informed risk management strategies and to use this knowledge to address ongoing problems posed by bovine spongiform encephalopathy (BSE), the gathering crisis of chronic wasting disease (CWD), emerging issues of human prion disease, and basic scientific understanding of the nature of prions. PrioNet is strategically responding to prion threats by focusing its network of highly accomplished researchers and trainees to implement integrated risk management strategies that could not be supported by other mechanisms.

Evaluating the use of house sparrow nestlings as sentinels for West Nile virus in Saskatchewan.

This study evaluated the use of house sparrow (Passer domesticus) nestlings as sentinels of West Nile virus (WNV) in the prairie grasslands of Saskatchewan. In the summer of 2006, 600 house sparrow nestlings were collected and pooled tissues tested by reverse transcriptase-polymerase chain reaction. All tested negative for WNV. During the same period, no WNV was detected by mosquito surveillance in the study area and 15 WNV-infected pools were collected from the nearby city of Estevan. Six percent of avian carcasses collected from Regina, a city 100 km from the study area in the same ecozone, were infected with WNV. In 2007, 200 house sparrow nestlings were collected and 4 tested positive for WNV, a prevalence of 2%. Ninety-seven house sparrow eggs were also collected and WNV antibodies were measured in the yolk. Seven eggs had measurable titers, a prevalence of 7.2%. Combined WNV surveillance showed high levels of WNV transmission in 2007; 112 WNV-infected mosquito pools were collected from nearby cities of Estevan and Weyburn, and the proportion of WNV infected avian carcasses from Regina was 78%. There were 1456 human cases of WNV in Saskatchewan in 2007, compared to 19 cases in 2006. The study concluded that house sparrow nestlings are not useful as an early warning of WNV circulation, or as a measure of the intensity of WNV activity in the prairie grasslands. Also, the study determined that maternally derived antibody did not have a significant limiting effect on WNV transmission to house sparrow nestlings in 2007, a year of epidemic WNV activity in the study area.

Multiple zoonotic pathogens identified in canine feces collected from a remote Canadian indigenous community.

Five genera of potentially zoonotic bacteria and parasites were detected in environmentally collected fecal samples from a remote indigenous community in Northern Saskatchewan, Canada. Organisms identified include Toxocara canis, Echniococcus granulosus, Giardia duodenalis, Cryptosporidium spp., and Campylobacter spp. The prevalence and intensity of Giardia spp. and Campylobacter spp. in fecal samples was particularly remarkable. Three-quarters of samples tested contained at least one zoonotic species of Campylobacter, and C. jejuni-containing feces had an average of 2.9 x 10(5) organisms/g. Over one-half of samples tested contained Giardia spp. with an average of 9,266 cysts/g. Zoonotic G. duodenalis Assemblage A was the only Giardia spp. genotype identified. These data suggest that canine feces have the potential to pose a significant health risk to Canadians in rural and remote indigenous communities.

Emergence of sylvatic Echinococcus granulosus as a parasitic zoonosis of public health concern in an indigenous community in Canada.

Within a remote Canadian Indigenous community, at least 11* of people had antibodies against Echinococcus granulosus and E. granulosus eggs were detected in 6* of environmentally collected canine fecal samples. Dog ownership, hunting, and trapping were not risk factors for seropositivity, suggesting that people are most likely exposed to E. granulosus through indirect contact with dog feces in the environment. In this situation, human exposure could be most effectively curtailed by preventing consumption of cervid viscera by free-roaming dogs.

Prionet Canada: a network of centres of excellence for research into prions and prion diseases.

PrioNet Canada's strength in basic, applied, and social research is helping to solve the food, health safety, and socioeconomic problems associated with prion diseases. Prion diseases are transmissible, fatal neurodegenerative diseases of humans and animals. Examples of prion diseases include bovine spongiform encephalopathy (BSE, commonly known as "mad cow" disease), Creutzfeldt-Jakob disease in humans, and chronic wasting disease (CWD) in deer and elk. As of March 31, 2008, PrioNet's interdisciplinary network included 62 scientific members, 5 international collaborators, and more than 150 students and young professionals working in partnership with 25 different government, nongovernment, and industry partners. PrioNet's activities are developing strategies based on a sustained, rational approach that will mitigate, and ultimately control, prion diseases in Canada.

Canada's inter-agency wild bird influenza survey.

Canada's Inter-agency Wild Bird Influenza Survey, which started in 2005, and is a collaborative effort among federal, provincial and territorial government agencies as well as non-governmental organizations and academic institutions, has two components: (i) a seasonal survey of live wild waterfowl species from selected geographic regions across Canada (to expand our understanding of the avian influenza viruses circulating in wild bird populations); and (ii) an ongoing survey of birds found dead that are submitted to a regional diagnostic laboratory (to enhance detection of highly pathogenic avian influenza strains). Combined swabs from the cloaca and oropharynx collected from each bird are screened using a real-time reverse transcriptase-polymerase chain reaction (RRT-PCR) that targets a unique segment of the influenza A M1 gene. If the M1 result is positive or inconclusive, RRT-PCR for gene segments of the H5 and H7 hemagglutinin subtypes are performed. All samples that are RRT-PCR positive for H5 or H7 are sent immediately for test confirmation and further characterization. All field and laboratory data are entered into a database developed and maintained by the Canadian Cooperative Wildlife Health Centre. Since the survey commenced in 2005, on average, 30% of all live ducks sampled, 5% of other species of live birds and 3% of birds found dead have tested positive for avian influenza, all of North American lineage and of low pathogenicity.

Wild bird influenza survey, Canada, 2005.

Of 4,268 wild ducks sampled in Canada in 2005, real-time reverse transcriptase-PCR detected influenza A matrix protein (M1) gene sequence in 37% and H5 gene sequence in 5%. Mallards accounted for 61% of samples, 73% of M1-positive ducks, and 90% of H5-positive ducks. Ducks hatched in 2005 accounted for 80% of the sample.

Diversity of Bartonella genotypes in Richardson's ground squirrel populations.

The diversity and dynamics of Bartonella genotypes found in wild Richardson's ground squirrels (RGS), Spermophilus richardsonii were monitored at multiple sites in Saskatchewan, Canada from 2002 to 2004. Based on sequence analysis of a portion of the Bartonella citrate synthase (gltA) gene, four different genotypes were detected in 233 isolates from 176 animals. The majority (87%) of sequences were identified as genotype H, with genotypes I, J, and K accounting for 8%, 4%, and 1% of sequences, respectively. Only one animal was concurrently infected with multiple Bartonella genotypes. Of 23 animals sampled four times or more, 26% were never infected with Bartonella. Of 32 RGS infected with Bartonella at first capture and then sampled again the following month, 50% were infected with the same Bartonella genotype, 41% were no longer infected, and 9% were infected with a different Bartonella genotype in the subsequent sample. The diversity of Bartonella genotypes varied among sites. At one site almost all RGS were infected with genotype H in September, and up to 60% of the same population was infected with genotype I the following spring. We compare our results with previous studies of Bartonella infections in rodents and discuss possible explanations for the observed differences.

Rodent-associated Bartonella in Saskatchewan, Canada.

Six species of wild rodents were sampled at 10 sites in 2002 and 2003 to determine the prevalence of Bartonella infections in rodent communities near Saskatoon, Saskatchewan, Canada. Isolates were characterized genotypically and compared with isolates found at other locations. Of 104 wild rodents examined, 57% were infected with Bartonella and prevalence within species varied from 49% for Richardson's ground squirrels (Spermophilus richardsonii) to 90% for Franklin's ground squirrels (S. franklinii). Infected rodents were found at all sites. Sequencing of a 379-bp portion of the citrate synthase gene was performed on 54 isolates and revealed 13 distinct genotypes, eight of which had not been described previously. The most common genotype detected in red-backed voles (Clethrionomys gapperi) was 99.1% similar to B. grahamii, a known human pathogen. Two of 10 Franklin's ground squirrels were concurrently infected with multiple Bartonella genotypes. All genotypes, with the exception of one detected in both Franklin's and thirteen-lined ground squirrels (S. tridecemlineatus), were found in only one host, and all genotypes from each species, with the exception of genotypes detected in red-backed voles, clustered together within the same relatedness group, suggesting that at least some Bartonella genotypes are specific to some rodent hosts.