Anticoagulant Rodenticide Toxicity in Terrestrial Raptors: Tools to Estimate the Impact on Populations in North America and Globally.

Anticoagulant rodenticides (ARs) have caused widespread contamination and poisoning of predators and scavengers. The diagnosis of toxicity proceeds from evidence of hemorrhage, and subsequent detection of residues in liver. Many factors confound the assessment of AR poisoning, particularly exposure dose, timing and frequency of exposure, and individual and taxon-specific variables. There is a need, therefore, for better AR toxicity criteria. To respond, we compiled a database of second-generation anticoagulant rodenticide (SGAR) residues in liver and postmortem evaluations of 951 terrestrial raptor carcasses from Canada and the United States, 1989 to 2021. We developed mixed-effects logistic regression models to produce specific probability curves of the toxicity of ∑SGARs at the taxonomic level of the family, and separately for three SGARs registered in North America, brodifacoum, bromadiolone, and difethialone. The ∑SGAR threshold concentrations for diagnosis of coagulopathy at 0.20 probability of risk were highest for strigid owls (15 ng g-1) lower and relatively similar for accipitrid hawks and eagles (8.2 ng g-1) and falcons (7.9 ng g-1), and much lower for tytonid barn owls (0.32 ng g-1). These values are lower than those we found previously, due to compilation and use of a larger database with a mix of species and source locations, and also to refinements in the statistical methods. Our presentation of results on the family taxonomic level should aid in the global applicability of the numbers. We also collated a subset of 440 single-compound exposure events and determined the probability of SGAR-poisoning symptoms as a function of SGAR concentration, which we then used to estimate relative SGAR toxicity and toxic equivalence factors: difethialone, 1, brodifacoum, 0.8, and bromadiolone, 0.5. Environ Toxicol Chem 2024;43:988-998. © 2024 His Majesty the King in Right of Canada and The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Assessing plastic ingestion in birds of prey from British Columbia, Canada.

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.

A novel calicivirus discovered in trumpeter swans (Cygnus buccinator) expands the richness of known avian caliciviruses.

Caliciviruses are ssRNA viruses that can infect a wide range of hosts, including birds. While several avian caliciviruses have been discovered, their taxonomy and host distribution are largely unknown. We molecularly characterized a novel calicivirus (trumpeter swan calicivirus: TruSCV) in trumpeter swans over-wintering in south-west British Columbia, Canada. The positivity rate was 20.3% (14/69) and there were no significant differences in infection rates between males (5/34, 14.7%) and females (9/35, 25.7%) or among considered age groups (juveniles: 4/14, 28.6%; sub-adults: 1/9, 11.1%; adults: 9/46, 19.6%). Twelve infected swans died of lead poisoning, one because of starvation, and one from physical injuries. TruSCV complete genome possessed the typical organization and protein motifs of caliciviruses and a type 2 IRES and its closest relative was a virus circulating in Australian ducks. Phylogenetic analyses showed the existence of 34 different but monophyletic avian caliciviruses. These viruses, while having conserved genomic organization and protein motifs, possess different IRES types and group in several divergent clades, with only two of them corresponding to currently defined genera, highlighting the need for epidemiological investigations and systematic analyses to better define their taxonomy. Follow-up studies are needed to elucidate the diversity, distribution, and pathogenic potential of TruSCV.

A threat from both sides: Multiple introductions of genetically distinct H5 HPAI viruses into Canada via both East Asia-Australasia/Pacific and Atlantic flyways.

From 2016 to 2020, high pathogenicity avian influenza (HPAI) H5 viruses circulated in Asia, Europe, and Africa, causing waves of infections and the deaths of millions of wild and domestic birds and presenting a zoonotic risk. In late 2021, H5N1 HPAI viruses were isolated from poultry in Canada and also retrospectively from a great black-backed gull (Larus marinus), raising concerns that the spread of these viruses to North America was mediated by migratory wild bird populations. In February and April 2022, H5N1 HPAI viruses were isolated from a bald eagle (Haliaeetus leucocephalus) and broiler chickens in British Columbia, Canada. Phylogenetic analysis showed that the virus from bald eagle was genetically related to H5N1 HPAI virus isolated in Hokkaido, Japan, in January 2022. The virus identified from broiler chickens was a reassortant H5N1 HPAI virus with unique constellation genome segments containing PB2 and NP from North American lineage LPAI viruses, and the remaining gene segments were genetically related to the original Newfoundland-like H5N1 HPAI viruses detected in November and December 2021 in Canada. This is the first report of H5 HPAI viruses' introduction to North America from the Pacific and the North Atlantic-linked flyways and highlights the expanding risk of genetically distinct virus introductions from different geographical locations and the potential for local reassortment with both the American lineage LPAI viruses in wild birds and with both Asian-like and European-like H5 HPAI viruses. We also report the presence of some amino acid substitutions across each segment that might contribute to the replicative efficiency of these viruses in mammalian host, evade adaptive immunity, and pose a potential zoonotic risk.

Anticoagulant Rodenticide Contamination of Terrestrial Birds of Prey from Western Canada: Patterns and Trends, 1988-2018.

As the dominant means for control of pest rodent populations globally, anticoagulant rodenticides (ARs), particularly the second-generation compounds (SGARs), have widely contaminated nontarget organisms. We present data on hepatic residues of ARs in 741 raptorial birds found dead or brought into rehabilitation centers in British Columbia, Canada, over a 30-year period from 1988 to 2018. Exposure varied by species, by proximity to residential areas, and over time, with at least one SGAR residue detected in 74% of individuals and multiple residues in 50% of individuals. By comparison, we detected first-generation compounds in <5% of the raptors. Highest rates of exposure were in barred owls (Strix varia), 96%, and great horned owls (Bubo virginianus), 81%, species with diverse diets, including rats (Rattus norvegicus and Rattus rattus), and inhabiting suburban and intensive agricultural habitats. Barn owls (Tyto alba), mainly a vole (Microtus) eater, had a lower incidence of exposure of 65%. Putatively, bird-eating raptors also had a relatively high incidence of exposure, with 75% of Cooper's hawks (Accipiter cooperii) and 60% of sharp-shinned hawks (Accipiter striatus) exposed. Concentrations of SGARs varied greatly, for example, in barred owls, the geometric mean ∑SGAR = 0.13, ranging from <0.005 to 1.81 µg/g wet weight (n = 208). Barred owls had significantly higher ∑SGAR concentrations than all other species, driven by significantly higher bromadiolone concentrations, which was predicted by the proportion of residential land within their home ranges. Preliminary indications that risk mitigation measures implemented in 2013 are having an influence on exposure include a decrease in mean concentrations of brodifacoum and difethialone in barred and great horned owls and an increase in bromodialone around that inflection point. Environ Toxicol Chem 2022;41:1903-1917. © 2022 Her Majesty the Queen in Right of Canada. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. Reproduced with the permission of the Minister of Environment and Climate Change Canada.

Seasonal variability in vulnerability for Cassin's auklets (Ptychoramphus aleuticus) exposed to microplastic pollution in the Canadian Pacific region.

Marine plastic pollution is an emerging global conservation challenge, potentially impacting organisms at all trophic levels. However, currently it is unclear to what extent plastic pollution is impacting marine organisms at the population, species or multispecies level. In this study, we explore seasonal exposure (i.e., vulnerability) of Cassin's Auklet (Ptychoramphus aleuticus) to plastic pollution with exposure models during boreal summer and winter seasons. Based on these models, we infer exposure at the population level for this species, in the Canadian Pacific region where approximately 75% of the global population of this species breeds. The models quantify plastic exposure by determining seasonal core foraging areas and plastic concentrations found in those same areas. Core foraging areas were determined using a Generalized Additive Model based on at-sea observation data (collected year round: 1990-2010) and 50% Home Range Kernels based on aerial telemetry data (May and June 1999-2001). Plastic concentrations within these core areas were interpolated based on seawater microplastic concentrations from the summer of 2012. We found that during the boreal summer, Cassin's Auklets were exposed to relatively low concentrations of plastics. During the winter, auklet distribution shifted towards the coast where plastic concentrations are considerably higher. Model derived seasonal variability in exposure was consistent with necropsy results from bird carcasses recovered during the winter of 2014, and from a multiyear study on chick provisioning during the summer. Local oceanography likely plays a role in determining seasonal shifts in both marine bird as well microplastic concentrations, and hence exposure. As well, individual sensitivity (i.e., dose-dependent effect) may vary with annual cycles. Currently, research is focusing on determining how sensitive individual birds are to microplastic concentrations, and our models will help translate sensitivity found at the individual level to potential impacts at population or species level.

Are ocean conditions and plastic debris resulting in a 'double whammy' for marine birds?

We report a mortality event of Red Phalaropes (Phalaropus fulicarius) that occurred from October to November 2016 on the north coast of British Columbia, Canada. All individuals were severely underweight and showing signs of physiological stress. The guts of all carcasses contained ingested plastics (100%, n = 9). Distribution modelling from pelagic bird surveys (1990-2010) indicated that Red Phalaropes are not typically found in the study area during fall months. Ocean conditions during fall 2016 were unusually warm, coinciding with reduced upwelling in the study area. eBird records since 1980 indicated Red Phalaropes are observed closer to shore during periods associated with reduced upwelling. These results suggest that distribution shifts of Red Phalaropes closer to shore, where plastic debris occurs in higher concentrations, may lead phalaropes to feed on plastic debris while in a weakened state, resulting in a combination of two adverse circumstances.

Increased rodenticide exposure rate and risk of toxicosis in barn owls (Tyto alba) from southwestern Canada and linkage with demographic but not genetic factors.

Among many anthropogenic drivers of population decline, continual rapid urbanization and industrialization pose major challenges for the survival of wildlife species. Barn owls (Tyto alba) in southwestern British Columbia (BC) face a multitude of threats ranging from habitat fragmentation to vehicle strikes. They are also at risk from secondary poisoning of second-generation anticoagulant rodenticides (SGARs), a suite of toxic compounds which at high doses results in a depletion of blood clotting factors leading to internal bleeding and death. Here, using long-term data (N = 119) for the hepatic residue levels of SGAR, we assessed the risk of toxicosis from SGAR for the BC barn owl population over the past two decades. We also investigated whether sensitivity to SGAR is associated with genetic factors, namely Single Nucleotide Polymorphisms (SNPs) found in the CYP2C45 gene of barn owls. We found that residue concentration for total SGAR was significantly higher in 2006-2013 (141 ng/g) relative to 1992-2003 (57 ng/g). The proportion of owls exposed to multiple SGAR types was also significantly higher in 2006-2013. Those measures accordingly translate directly into an increase in toxicosis risk level. We also detected demographic differences, where adult females showed on average lower concentration of total SGAR (64 ng/g) when compared to adult males (106 ng/g). Juveniles were overall more likely to show signs of toxicosis than adults (33.3 and 6.9 %, respectively), and those symptoms were positively predicted by SGAR concentrations. We found no evidence that SNPs in the CYP2C45 gene of barn owls were associated with intraspecific variation in SGAR sensitivity. We recommend several preventative measures be taken to minimize wildlife exposure to SGAR.

Pathobiological Characterization of a Novel Reassortant Highly Pathogenic H5N1 Virus Isolated in British Columbia, Canada, 2015.

In the current study, we describe the pathobiologic characteristics of a novel reassortant virus - A/chicken/BC/FAV-002/2015 (H5N1) belonging to clade 2.3.4.4 that was isolated from backyard chickens in British Columbia, Canada. Sequence analyses demonstrate PB1, PA, NA and NS gene segments were of North American lineage while PB2, HA, NP and M were derived from a Eurasian lineage H5N8 virus. This novel virus had a 19 amino acid deletion in the neuraminidase stalk. We evaluated the pathogenic potential of this isolate in various animal models. The virus was highly pathogenic to mice with a LD50 of 10 plaque forming units (PFU), but had limited tissue tropism. It caused only subclinical infection in pigs which did result in seroconversion. This virus was highly pathogenic to chickens, turkeys, juvenile Muscovy ducks (Cairnia moschata foma domestica) and adult Chinese geese (Anser cynoides domesticus) causing a systemic infection in all species. The virus was also efficiently transmitted and resulted in mortality in naïve contact ducks, geese and chickens. Our findings indicate that this novel H5N1 virus has a wide host range and enhanced surveillance of migratory waterfowl may be necessary in order to determine its potential to establish itself in the wild bird reservoir.

Reassortant highly pathogenic influenza A H5N2 virus containing gene segments related to Eurasian H5N8 in British Columbia, Canada, 2014.

In late November 2014 higher than normal death losses in a meat turkey and chicken broiler breeder farm in the Fraser Valley of British Columbia initiated a diagnostic investigation that led to the discovery of a novel reassortant highly pathogenic avian influenza (HPAI) H5N2 virus. This virus, composed of 5 gene segments (PB2, PA, HA, M and NS) related to Eurasian HPAI H5N8 and the remaining gene segments (PB1, NP and NA) related to North American lineage waterfowl viruses, represents the first HPAI outbreak in North American poultry due to a virus with Eurasian lineage genes. Since its first appearance in Korea in January 2014, HPAI H5N8 spread to Western Europe in November 2014. These European outbreaks happened to temporally coincide with migratory waterfowl movements. The fact that the British Columbia outbreaks also occurred at a time associated with increased migratory waterfowl activity along with reports by the USA of a wholly Eurasian H5N8 virus detected in wild birds in Washington State, strongly suggest that migratory waterfowl were responsible for bringing Eurasian H5N8 to North America where it subsequently reassorted with indigenous viruses.

Northern fulmars as biological monitors of trends of plastic pollution in the eastern North Pacific.

Marine plastic debris is a global issue, which highlights the need for internationally standardized methods of monitoring plastic pollution. The stomach contents of beached northern fulmar (Fulmarus glacialis) have proven a cost-effective biomonitor in Europe. However, recent information on northern fulmar plastic ingestion is lacking in the North Pacific. We quantified the stomach contents of 67 fulmars from beaches in the eastern North Pacific in 2009-2010 and found that 92.5% of fulmars had ingested an average of 36.8 pieces, or 0.385 g of plastic. Plastic ingestion in these fulmars is among the highest recorded globally. Compared to earlier studies in the North Pacific, our findings indicate an increase in plastic ingestion over the past 40 years. This study substantiates the use of northern fulmar as biomonitors of plastic pollution in the North Pacific and suggests that the high levels of plastic pollution in this region warrant further monitoring.

Investigation of the concurrent emergence of Salmonella enteritidis in humans and poultry in British Columbia, Canada, 2008-2010.

An increase in the rate of human infections with Salmonella enteritidis (SE) occurred between 2007 and 2010 in British Columbia (BC). During the same time period, an increase in SE from poultry-sourced isolates and increased clinical severity in poultry were also observed in BC. This article describes a multi-sectoral collaboration during a 3-year investigation, and the actions taken by public health and animal health professionals. Human cases were interviewed, clusters were investigated, and a case-control study was conducted. Environmental investigations were conducted in food service establishments (FSE). Suspect foods were tested. Laboratory data from poultry-sourced isolates were analysed. Five hundred and eighty-four human cases of SE with the same pulsed-field gel electrophoresis pattern were identified between May 2008 and August 2010. Seventy-three percentage of cases reported consumption of eggs. The odds of egg consumption were 2.4 times higher for cases than controls. Implicated FSE were found to use ungraded eggs, which had been distributed illegally. Investigation suggested that there were multiple suppliers of these eggs. Collaboration between public health and animal health professionals led to data sharing, improved understanding of SE, engagement with the poultry industry and public communication. Multi-disciplinary, multi-sectoral and multi-pronged investigations are recommended to identify the likely source of illness in large, protracted foodborne outbreaks caused by commonly consumed foods.

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.

After the outbreak: how the British Columbia commercial poultry industry recovered after H7N3 HPAI.

In spring 2004, an outbreak of highly pathogenic avian influenza (HPAI), subtype H7N3, occurred in the Fraser Valley of British Columbia, Canada. The active outbreak lasted more than 90 days; 42 commercial poultry farms were identified as infected premises, and more than 17 million birds were culled. Through the depopulation of HPAI-positive farms and the strategic depopulation of adjacent test-negative farms, a total of 410 commercial poultry farms were emptied. The goals for the commercial poultry industry were to expedite restocking, reduce nonproductive downtime, negotiate equitable financial compensation, review and restructure emergency disease response plans, and identify and implement mitigation strategies. After the outbreak, multijurisdictional reviews identified the strengths and weaknesses of the outbreak control strategy. Lessons learned were incorporated into current emergency disease response protocols for both industry and government. The industry-led challenge to initial compensation values, especially for specialty poultry and breeder birds, resulted in a review of the federal Health of Animals Act. The British Columbia poultry industry, in collaboration with the British Columbia Ministry of Agriculture and Lands, developed an Enhanced Biosecurity Initiative that included the identification of mandatory on-farm biosecurity standards for commercial producers, an educational biosecurity self-assessment guide, and provisions for a producer self-quarantine to be enacted upon the first suspicion of disease.

Relationship between H5N2 avian influenza viruses isolated from wild and domestic ducks in British Columbia, Canada.

In the summer of 2005 a Canadian national surveillance program for influenza A viruses in wild aquatic birds was initiated. The program involved collaboration between federal and provincial levels of government and was coordinated by the Canadian Cooperative Wildlife Health Centre. The surveillance plan targeted young-of-the-year Mallards along with other duck species at six sampling locations along the major migratory flyways across Canada. Beginning in early August, cloacal swabs were taken from 704 ducks on two lakes adjacent to one another near Kamloops, British Columbia. The swabs were screened for the presence of influenza A RNA using a real-time reverse transcription-polymerase chain reaction (RRT-PCR) assay that targets the M1 gene. Swab samples that gave positive results underwent further testing using H5- and H7-specific RRT-PCR assays. One hundred and seventy-four cloacal swab specimens gave positive or suspicious results for the presence of an H5 virus. A portion of these (28/35) were confirmed using an H5-specific conventional reverse transcription-polymerase chain reaction assay and an H5 virus was eventually isolated from 24/127 swab specimens. Neuraminidase typing revealed the presence of H5N2 and H5N9 viruses. In mid-November of 2005 an H5N2 virus was detected in a commercial duck operation in the lower mainland of British Columbia, approximately 120 km from where the H5N2-positive wild ducks were sampled. Molecular genetic analysis of the H5N2 viruses isolated from wild and domestic ducks was carried out to determine their kinship.

Novel avian influenza H7N3 strain outbreak, British Columbia.

Genome sequences of chicken (low pathogenic avian influenza [LPAI] and highly pathogenic avian influenza [HPAI]) and human isolates from a 2004 outbreak of H7N3 avian influenza in Canada showed a novel insertion in the HA0 cleavage site of the human and HPAI isolate. This insertion likely occurred by recombination between the hemagglutination and matrix genes in the LPAI virus.