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Following the detection of novel highly pathogenic avian influenza virus (HPAIV) H5N1 clade 2.3.4.4b in Newfoundland, Canada, in late 2021, avian influenza virus (AIV) surveillance in wild birds was scaled up across Canada. Herein, we present the results of Canada's Interagency Surveillance Program for Avian Influenza in Wild Birds during the first year (November 2021-November 2022) following the incursions of HPAIV from Eurasia. The key objectives of the surveillance program were to (i) identify the presence, distribution, and spread of HPAIV and other AIVs; (ii) identify wild bird morbidity and mortality associated with HPAIV; (iii) identify the range of wild bird species infected by HPAIV; and (iv) genetically characterize detected AIV. A total of 6,246 sick and dead wild birds were tested, of which 27.4% were HPAIV positive across 12 taxonomic orders and 80 species. Geographically, HPAIV detections occurred in all Canadian provinces and territories, with the highest numbers in the Atlantic and Central Flyways. Temporally, peak detections differed across flyways, though the national peak occurred in April 2022. In an additional 11,295 asymptomatic harvested or live-captured wild birds, 5.2% were HPAIV positive across 3 taxonomic orders and 19 species. Whole-genome sequencing identified HPAIV of Eurasian origin as most prevalent in the Atlantic Flyway, along with multiple reassortants of mixed Eurasian and North American origins distributed across Canada, with moderate structuring at the flyway scale. Wild birds were victims and reservoirs of HPAIV H5N1 2.3.4.4b, underscoring the importance of surveillance encompassing samples from sick and dead, as well as live and harvested birds, to provide insights into the dynamics and potential impacts of the HPAIV H5N1 outbreak. This dramatic shift in the presence and distribution of HPAIV in wild birds in Canada highlights a need for sustained investment in wild bird surveillance and collaboration across interagency partners., Importance: We present the results of Canada's Interagency Surveillance Program for Avian Influenza in Wild Birds in the year following the first detection of highly pathogenic avian influenza virus (HPAIV) H5N1 on the continent. The surveillance program tested over 17,000 wild birds, both sick and apparently healthy, which revealed spatiotemporal and taxonomic patterns in HPAIV prevalence and mortality across Canada. The significant shift in the presence and distribution of HPAIV in Canada's wild birds underscores the need for sustained investment in wild bird surveillance and collaboration across One Health partners., Competing Interests: The authors declare no conflict of interest.

Highly pathogenic avian influenza (HPAI) viruses have spread at an unprecedented scale, leading to mass mortalities in birds and mammals. In 2023, a transatlantic incursion of HPAI A(H5N5) viruses into North America was detected, followed shortly thereafter by a mammalian detection. As these A(H5N5) viruses were similar to contemporary viruses described in Eurasia, the transatlantic spread of A(H5N5) viruses was most likely facilitated by pelagic seabirds. Some of the Canadian A(H5N5) viruses from birds and mammals possessed the PB2-E627K substitution known to facilitate adaptation to mammals. Ferrets inoculated with A(H5N5) viruses showed rapid, severe disease onset, with some evidence of direct contact transmission. However, these viruses have maintained receptor binding traits of avian influenza viruses and were susceptible to oseltamivir and zanamivir. Understanding the factors influencing the virulence and transmission of A(H5N5) in migratory birds and mammals is critical to minimize impacts on wildlife and public health., Competing Interests: Declaration of interests The authors declare no competing interests., (Crown Copyright © 2024. Published by Elsevier Inc. All rights reserved.)

The GsGd lineage (A/goose/Guangdong/1/1996) H5N1 virus was introduced to Canada in 2021/2022 through the Atlantic and East Asia-Australasia/Pacific flyways by migratory birds. This was followed by unprecedented outbreaks affecting domestic and wild birds, with spillover into other animals. Here, we report sporadic cases of H5N1 in 40 free-living mesocarnivore species such as red foxes, striped skunks, and mink in Canada. The clinical presentations of the disease in mesocarnivores were consistent with central nervous system infection. This was supported by the presence of microscopic lesions and the presence of abundant IAV antigen by immunohistochemistry. Some red foxes that survived clinical infection developed anti-H5N1 antibodies. Phylogenetically, the H5N1 viruses from the mesocarnivore species belonged to clade 2.3.4.4b and had four different genome constellation patterns. The first group of viruses had wholly Eurasian (EA) genome segments. The other three groups were reassortant viruses containing genome segments derived from both North American (NAm) and EA influenza A viruses. Almost 17 percent of the H5N1 viruses had mammalian adaptive mutations (E627 K, E627V and D701N) in the polymerase basic protein 2 (PB2) subunit of the RNA polymerase complex. Other mutations that may favour adaptation to mammalian hosts were also present in other internal gene segments. The detection of these critical mutations in a large number of mammals within short duration after virus introduction inevitably highlights the need for continually monitoring and assessing mammalian-origin H5N1 clade 2.3.4.4b viruses for adaptive mutations, which potentially can facilitate virus replication, horizontal transmission and posing pandemic risks for humans.

We identified by light microscopy micro- and macrogametes and oocysts of renal coccidia in 78 of 220 (35.5%) Northern Gannets (Morus bassanus) from the western North Atlantic population. This infection was not considered clinically significant in any of the affected birds, although the potential effect of this parasite in breeding colonies, particularly among nestlings, is unknown. Analysis of the 18S rRNA gene from frozen renal tissue by PCR and subsequent sequencing revealed 95.6% identity with Eimeria auritusi from Double-crested Cormorants (Phalacrocorax auritus), suggesting a novel Eimeria sp. in the Northern Gannets., (© Wildlife Disease Association 2023.)

In summer 2022, highly pathogenic influenza A(H5N1) virus reached the herring gull (Larus argentatus subspecies smithsonianus) breeding colony on Kent Island, New Brunswick, Canada. Real-time monitoring revealed a self-limiting outbreak with low mortality. Proactive seabird surveillance is crucial for monitoring such limited outbreaks, protecting seabirds, and tracing zoonotic transmission routes.

In December 2022 and January 2023, we isolated clade 2.3.4.4b H5N1 high-pathogenicity avian influenza (HPAI) viruses from six American crows ( Corvus brachyrhynchos ) from Prince Edward Island and a red fox ( Vulpes vulpes ) from Newfoundland, Canada. Using full-genome sequencing and phylogenetic analysis, these viruses were found to fall into two distinct phylogenetic clusters: one group containing H5N1 viruses that had been circulating in North and South America since late 2021, and the other one containing European H5N1 viruses reported in late 2022. The transatlantic re-introduction for the second time by pelagic/Icelandic bird migration via the same route used during the 2021 incursion of Eurasian origin H5N1 viruses into North America demonstrates that migratory birds continue to be the driving force for transcontinental dissemination of the virus. This new detection further demonstrates the continual long-term threat of H5N1 viruses for poultry and mammals and the subsequent impact on various wild bird populations wherever these viruses emerge. The continual emergence of clade 2.3.4.4b H5Nx viruses requires vigilant surveillance in wild birds, particularly in areas of the Americas, which lie within the migratory corridors for long-distance migratory birds originating from Europe and Asia. Although H5Nx viruses have been detected at higher rates in North America since 2021, a bidirectional flow of H5Nx genes of American origin viruses to Europe has never been reported. In the future, coordinated and systematic surveillance programs for HPAI viruses need to be launched between European and North American agencies.

Objective: Molecular identification of small cestodes, morphologically consistent with Echinococcus multilocularis , recovered at necropsy from the gastrointestinal tract contents of a red fox, was accomplished by PCR using published species-specific n ad1 primers and methods., Animal: Red fox (Vulpes vulpes) ., Procedure: Small cestodes recovered from intestinal contents of a red fox trapped on Prince Edward Island in December 2020 (frozen at -20°C before being processed for parasite recovery in June 2021) were morphologically identified. Species identity confirmation and haplotyping of the cestodes were done via PCR and DNA sequencing of the n ad1, nad2 , and cob genes., Results: Small cestodes morphologically consistent with E. multilocularis were detected in the gastrointestinal tract contents of a red fox trapped near Montague, PEI. The species identity was confirmed via PCR. Haplotyping revealed that they were of the European E1 haplotype., Conclusion: In Canada, E. multilocularis has been reported as far east as Québec, with most reports being in central and western provinces and territories. This is the first report of E. multilocularis infection in a canid host east of Ontario, Canada and illustrates the need for regular wildlife disease surveillance to enhance our understanding of emerging pathogens of veterinary and medical importance., Clinical Relevance: Echinococcus multilocularis is a highly pathogenic zoonotic cestode from the family Taeniidae that can cause alveolar echinococcosis (AE) when rodents, dogs, horses, pigs, non-human primates, or humans ingest its eggs. Alveolar echinococcosis is challenging to treat, and survival rates for untreated individuals are low., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

Objective: To identify first-stage nematode larvae (L1) recovered from a red fox scat sample and adult female worms recovered from 2 red fox lungs at necropsy, using published molecular methods to confirm a morphological diagnosis of Angiostrongylus vasorum (French heartworm)., Animal: Red fox (Vulpes vulpes) ., Procedure: Nematode larvae recovered from a Baermann examination survey of wild canid scats ( n = 101) conducted from January 2017 to August 2020, were identified by size and morphology and subjected to PCR and DNA sequencing of the small subunit (SSU) rRNA gene, the large subunit (LSU) rRNA gene, or the second internal transcribed spacer (ITS2). In addition, these techniques were applied to adult female worms recovered from the heart/lungs of 2 red foxes (obtained from PEI trappers and stored frozen at -20°C since December of 2018 and 2020)., Results: Size and morphology of L1 recovered by Baermann examination from a wild canid scat sample (presumed to be red fox) collected near Montague, PEI and adult female worms recovered at necropsy from 2 red fox carcasses were identified as A. vasorum . Molecular analysis confirmed the larvae and adult worms were A. vasorum ., Conclusion: These findings indicated that A. vasorum has become endemic in the red fox population on PEI., Clinical Relevance: Angiostrongylus vasorum infection is potentially fatal in dogs. Veterinarians and regional diagnostic laboratories in the Maritime provinces should consider the possibility of A. vasorum infection in dogs with clinical signs of cardiopulmonary and/or central nervous system disease or bleeding disorders., (Copyright and/or publishing rights held by the Canadian Veterinary Medical Association.)

Skunk adenovirus-1 (SkAdV-1) has been reported infecting several North American wildlife species; however, lesions associated with disease have not yet been completely characterized, particularly in porcupines. We describe and characterize the tissue distribution and lesions associated with SkAdV-1 infection in 24 wildlife diagnostic cases submitted between 2015 and 2020, including 16 North American porcupines (Erethizon dorsatum), three striped skunks (Mephitis mephitis), and five raccoons (Procyon lotor), which constitute a new host species. The most common lesion in all species was severe necrotizing bronchopneumonia with (n=12) or without (n=10) interstitial involvement. Intranuclear inclusion bodies were common in respiratory epithelium (n=21) and less often in renal tubular (n=6) and biliary epithelium (n=1). Several cases (n=4) had secondary bacterial infections, including Bordetella bronchiseptica, Pasteurella multocida, and Streptococcus zooepidemicus. In situ hybridization in porcupine (n=6), raccoon (n=1), and skunk (n=1) revealed SkAdV-1 DNA in multiple tissue types, including lung, trachea, turbinates, liver, kidney, lymph node, and brain, and multiple cell types including epithelial, endothelial, and mesothelial cells. These findings were consistent across species. Comparison of viral genomes from a porcupine and a raccoon with that originally isolated from a skunk demonstrated DNA point mutations affecting several viral genes, including the fiber protein gene. Our findings show the spectrum of disease associated with SkAdV-1 infection in a broad host range of wildlife species., (© Wildlife Disease Association 2022.)

The Northern Gannet (Morus bassanus) is a large marine bird whose whole North American population breeds in waters of eastern Canada. Opportunities to identify causes of morbidity and mortality in recently hatched birds of this species are therefore limited to this region of North America. During the three decades since 1990 of wildlife health surveillance at the Atlantic regional center of the Canadian Wildlife Health Cooperative, what appears to be a previously undescribed syndrome of ischemic leg necrosis affecting mainly hatch-year Northern Gannets has emerged, which may relate to some unique aspects of the life history of these birds. This syndrome, observed in 14 birds, is characterized by severe necrosis and fibrinopurulent inflammation of soft tissues of the feet extending along the whole tarsometatarsus. An infectious cause is proposed to explain the pattern of lesions observed in these birds, possibly favored by a specialized and rich vascular system in their legs and feet. An acute or subacute cardiomyopathy, thought to be secondary to the severe leg lesions, was also observed microscopically in six of these birds., (© Wildlife Disease Association 2021.)

Purpose: To assess the comparative accuracy of commonly utilised index radiological measurements in the prognosis of infantile idiopathic scoliosis (IIS) and build a parsimonious prognostic model utilising these measurements., Methods: This was a retrospective analysis of a UK population of patients with IIS. Index radiological parameters were analysed, and outcome of their condition was determined over long-term follow-up. Comparative accuracy of each radiological measurement category was determined by logistic regression analyses and the corresponding receiver operating characteristic (ROC) curve. A predictive model of IIS progression using these measurements was then created., Results: All three radiological measurement categories were predictive of IIS progression. However, on pairwise comparison of ROC curves and multivariate analysis, the index Cobb angle proved the most significant predictor of curve progression. Using the index Cobb angle only, a predictive model of curve progression achieved an accuracy of 81.18% with a cut-off Cobb angle of 34.5° found to be the optimal threshold to discriminate a progressive from resolving curve., Conclusion: Of the three analysed index radiological parameters commonly used by surgeons in the prognosis of IIS, we found that the Cobb angle is the most accurate predictive measure. Further, neither addition of the RVAD nor convex RVA provided significant further prognostic value in a multivariate model of progression. However, not even the Cobb angle model was accurate in all cases; emphasising caution should be applied when relying on index radiological measurements to predict IIS outcomes., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.)

Purpose: Since its original description by Mehta, the rib vertebra angle difference (RVAD) and, in particular, a threshold of 20° have become an accepted and widely utilised prognostic indicator in the assessment of patients presenting with an infantile idiopathic scoliosis (IIS). However, uncertainty in the utility of the RVAD in the prognosis of IIS remains. The aims of this study were to investigate the prognostic significance of the RVAD and to describe the changes in RVAD over long-term follow-up of patients with progressive and resolving IIS., Methods: This was a retrospective analysis of patients presenting with IIS at a tertiary spinal deformity unit in the UK. Serial patient radiographs were reviewed and a logistic regression model using the patients index RVAD was created to predict the likelihood of curve progression., Results: At both index presentation and over long-term follow-up, patients with a progressive curve had significantly greater mean Cobb angle and RVAD measurements than those with resolving curves. The RVAD and Cobb were found to correlate positively in both groups, reflecting the underlying costovertebral pathoanatomy. The logistic regression model demonstrated that the optimal RVAD threshold in predicting IIS progression was lower at 17.1° than the 20° cut-off previously advocated., Conclusion: This study describes the utility of the RVAD in predicting IIS evolution. From this analysis, we would advise caution in predicting outcomes based on the index RVAD at presentation., Level of Evidence: II.

Egyptian rousette bats ( Rousettus aegyptiacus ) are natural reservoir hosts of Marburg virus (MARV), and Ravn virus (RAVV; collectively called marburgviruses) and have been linked to human cases of Marburg virus disease (MVD). We investigated the clinical and pathologic effects of experimental MARV infection in Egyptian rousettes through a serial euthanasia study and found clear evidence of mild but transient disease. Three groups of nine, captive-born, juvenile male bats were inoculated subcutaneously with 10,000 TCID 50 of Marburg virus strain Uganda 371Bat2007, a minimally passaged virus originally isolated from a wild Egyptian rousette. Control bats ( n = 3) were mock-inoculated. Three animals per day were euthanized at 3, 5⁻10, 12 and 28 days post-inoculation (DPI); controls were euthanized at 28 DPI. Blood chemistry analyses showed a mild, statistically significant elevation in alanine aminotransferase (ALT) at 3, 6 and 7 DPI. Lymphocyte and monocyte counts were mildly elevated in inoculated bats after 9 DPI. Liver histology revealed small foci of inflammatory infiltrate in infected bats, similar to lesions previously described in wild, naturally-infected bats. Liver lesion severity scores peaked at 7 DPI, and were correlated with both ALT and hepatic viral RNA levels. Immunohistochemical staining detected infrequent viral antigen in liver (3⁻8 DPI, n = 8), spleen (3⁻7 DPI, n = 8), skin (inoculation site; 3⁻12 DPI, n = 20), lymph nodes (3⁻10 DPI, n = 6), and oral submucosa (8⁻9 DPI, n = 2). Viral antigen was present in histiocytes, hepatocytes and mesenchymal cells, and in the liver, antigen staining co-localized with inflammatory foci. These results show the first clear evidence of very mild disease caused by a filovirus in a reservoir bat host and provide support for our experimental model of this virus-reservoir host system.