American black bears (Ursus americanus) can be considered vertebrate pests when they come in conflict with humans and create potential public safety situations that require intervention by local wildlife or animal control departments. Black bears are both omnivorous and highly intelligent, and this combination of characteristics make these animals prone to human habituation and developing an association between humans and easy sources of food. Foraging behaviors can put black bears in undesired or unsafe contact with humans when bears root through garbage on private and public lands, consume crops, prey upon small livestock or pets, cause significant property damage, and wander onto roadways leading to traffic accidents. An increasingly recognized cause of bear-human conflict is neurologic disease, such as that caused by infectious pathogens or toxicity, leading to cognitive or behavioral changes. A unique neurologic disease affecting black bears was first observed in 2014 near Lake Tahoe on the border between California and Nevada. Since then, it has been reported in eight counties in California and two counties in Nevada. Affected bears exhibit a range of neurologic symptoms including tremors, seizures, head tilts, and loss of fear of humans (i.e., “dog-like” behavior). Affected animals are typically young (<3 years old), underweight, and most often present in the spring after early emergence from hibernation. Histopathologic examination of brain tissue from these bears revealed varying degrees of inflammation in the brain (encephalitis), but the cause of this encephalitis was not apparent. Testing for common infectious causes of encephalitis (e.g., rabies, canine distemper virus, canine adenovirus, West Nile virus) and common neurotoxins did not identify a cause of the inflammation. A novel gammaherpesvirus, circovirus, parvovirus, and anellovirus were isolated from subsets of affected bears; however, a direct correlation between viral infection and encephalitis has not been established to date. The goal of this study is to investigate the potential role of protozoal parasites in this encephalitis of unknown origin, as members of this group of pathogens are known causes of encephalitis in humans and animals. Brain tissue from bears with encephalitis (n = 21) and unaffected bears (n = 15) from California and Nevada was screened via PCR targeting two loci commonly used to identify protozoal parasites (ITS1 and 18S). Protozoal parasite DNA was detected in 47.6% of bears with encephalitis (10/21) and 26.7% of unaffected bears (4/15). Parasites detected in bears with encephalitis included Sarcocystis neurona (4/21), S. canis (1/21), S. felis (1/21), an uncharacterized Sarcocystis sp. (1/21), Toxoplasma gondii (1/21), and an uncharacterized Cystoisospora-like species (2/21). Sarcocystis neurona, S. canis, and T. gondii are known to cause encephalitis in humans and animals. Sarcocystis canis is an increasingly recognized pathogen of bears and has been reported to cause inflammation in the brain, liver, and muscle in other geographical regions. The clinical significance of the poorly characterized protozoal species is uncertain. Parasites detected in unaffected bears included T. gondii (1/15), an uncharacterized Sarcocystis species (1/15) and an uncharacterized Cystoisospora-like species (2/15). These results suggest that protozoal parasites, particularly Sarcocystis species, may contribute to encephalitis in juvenile black bears, creating a public safety hazard at the human-wildlife interface.

From 2014-2019, 8 juvenile black bears (Ursus americanus) from different geographic regions were presented to the California Department of Fish and Wildlife because of emaciation, alopecia, and exfoliative dermatitis that resulted in death or euthanasia. Autopsy and histopathology revealed that all 8 bears had generalized hyperkeratotic dermatitis, folliculitis, and furunculosis. Skin structures were heavily colonized by fungal hyphae and arthrospores; fungal cultures of skin from 7 bears yielded Trichophyton equinum, a zoophilic dermatophyte reported only rarely in non-equid species. Additional skin conditions included mites (5), ticks (2), and coagulase-negative Staphylococcus sp. infections (2). No other causes of morbidity or mortality were identified. Molecular comparisons performed at the University of Texas Fungal Reference Laboratory determined that all isolates produced identical banding patterns, potentially representing a clonal population. Dermatophytosis is commonly localized and limited to the stratum corneum of the epidermis and hair follicles. Generalized disease with dermal involvement is rare in immunocompetent individuals; illness, malnutrition, age, or immunosuppression may increase susceptibility. Underlying causes for the severe disease impact in these bears were not evident after physical or postmortem examination. The mechanism by which bears from different geographic locations had severe, T. equinum-associated dermatophytosis from a potentially clonal dermatophyte could not be explained and warrants further investigation.

Since 2002, reports of deer with swollen muzzles from throughout the United States have resulted in significant interest by wildlife biologists and wildlife enthusiasts. The condition was identified in 25 white-tailed deer (Odocoileus virginianus) and 2 mule deer (O. hemionus). Microscopic lesions consisted of severe, granulomatous or pyogranulomatous inflammation of the muzzle, nasal planum, and upper lip, as well as similar but less severe inflammation of the hard palate. Lymphadenitis of regional lymph nodes was common and granulomatous pneumonia was present in one individual. Splendore-Hoeppli material was typical in the center of inflammatory foci. Other than the single instance of pneumonia, systemic disease was not evident. Various bacterial species were isolated in culture, most of which were not morphologically consistent with the colonies of small, gram-negative bacteria observed in the center of the granulomas. Amplification and sequencing of the bacterial 16S rRNA gene from tissues of affected deer resulted in the identification of Mannheimia granulomatis. Laser capture microdissection was used to confirm that the colonies in the inflammatory foci were M. granulomatis. The cases described here are reminiscent of a bovine disease in Brazil and Argentina, locally called lechiguana. Although the inflammation of lechiguana is mostly truncal, the microscopic lesions are very similar and are also attributed to M. granulomatis. It is unclear if this is an emerging infectious disease of deer, or if it is a sporadic, uncommon condition that has only recently been recognized.

Listeria monocytogenes is a bacterium that can cause disease in many species, including humans, livestock, and wildlife. Increased interactions via shared habitats may promote pathogen transmission among these groups. Our objectives were to evaluate the Southeastern Cooperative Wildlife Disease Study diagnostic data to characterize and compare L. monocytogenes-induced lesions and comorbidities in gray foxes and wild turkeys, and to describe cases of listeriosis in 2 cervids. From 1991-2020, 8 gray foxes, 8 wild turkeys, a neonatal elk, and a white-tailed deer fawn from several eastern states in the United States were diagnosed with listeriosis. All 8 foxes had hepatitis and/or hepatic necrosis with intralesional gram-positive bacilli, and concurrent canine distemper virus (CDV) infection; 2 of the foxes had been vaccinated recently for CDV. L. monocytogenes was cultured from the liver (6 of 8) or lung (2 of 8) of foxes. Lesions in wild turkeys included hepatocellular necrosis (3 of 8), heterophilic hepatitis (1 of 8), heterophilic granulomas (1 of 8), intrasinusoidal gram-positive bacilli without hepatic lesions (1 of 8), granulomatous dermatitis (1 of 8), and/or granulomatous myocarditis (2 of 8). Lymphoproliferative disease viral DNA was detected in 5 of 6 turkeys tested; reticuloendotheliosis viral DNA was detected in 2 of 3 turkeys tested. Both cervids had systemic listeriosis, with L. monocytogenes isolated from liver. Immunohistochemistry for Listeria spp. on select cases revealed immunolabeling in affected organs. Listeriosis was thus established as a cause of morbidity and mortality in 3 wildlife species, which often suffered from concurrent infections and likely immunosuppression.

Babesiosis is a potentially fatal tick-borne zoonotic disease caused by a species complex of blood parasites that can infect a variety of vertebrates, particularly dogs, cattle, and humans. In the United States, human babesiosis is caused by two distinct parasites, Babesia microti and Babesia duncani. The enzootic cycle of B. microti, endemic in the northeastern and upper midwestern regions, has been well characterised. In the western United States, however, the natural reservoir host and tick vector have not been identified for B. duncani, greatly impeding efforts to understand and manage this zoonotic disease. Two and a half decades after B. duncani was first described in a human patient in Washington State, USA, we provide evidence that the enzootic tick vector is the winter tick, Dermacentor albipictus, and the reservoir host is likely the mule deer, Odocoileus hemionus. The broad, overlapping ranges of these two species covers a large portion of far-western North America, and is consistent with confirmed cases of B. duncani in the far-western United States.

Viral infections were investigated in American black bears (Ursus americanus) from Nevada and northern California with and without idiopathic encephalitis. Metagenomics analyses of tissue pools revealed novel viruses in the genera Circoviridae, Parvoviridae, Anelloviridae, Polyomaviridae, and Papillomaviridae. The circovirus and parvovirus were of particular interest due to their potential importance as pathogens. We characterized the genomes of these viruses and subsequently screened bears by PCR to determine their prevalence. The circovirus (Ursus americanus circovirus, UaCV) was detected at a high prevalence (10/16, 67%), and the chaphamaparvovirus (Ursus americanus parvovirus, UaPV) was found in a single bear. We showed that UaCV is present in liver, spleen/lymph node, and brain tissue of selected cases by in situ hybridization (ISH) and PCR. Infections were detected in cases of idiopathic encephalitis and in cases without inflammatory brain lesions. Infection status was not clearly correlated with disease, and the significance of these infections remains unclear. Given the known pathogenicity of a closely related mammalian circovirus, and the complex manifestations of circovirus-associated diseases, we suggest that UaCV warrants further study as a possible cause or contributor to disease in American black bears.

Lymphoproliferative disease virus (LPDV) is an exogenous oncogenic retrovirus that induces lymphoid tumors in some galliform species of birds. Historically, outbreaks of LPDV have been reported from Europe and Israel. Although the virus has previously never been detected in North America, herein we describe the widespread distribution, genetic diversity, pathogenesis, and evolution of LPDV in the United States. Characterization of the provirus genome of the index LPDV case from North America demonstrated an 88% nucleotide identity to the Israeli prototype strain. Although phylogenetic analysis indicated that the majority of viruses fell into a single North American lineage, a small subset of viruses from South Carolina were most closely related to the Israeli prototype. These results suggest that LPDV was transferred between continents to initiate outbreaks of disease. However, the direction (New World to Old World or vice versa), mechanism, and time frame of the transcontinental spread currently remain unknown.

Lymphoproliferative disease virus (LPDV) is an exogenous oncogenic retrovirus that induces lymphoid tumors in some galliform species of birds. Historically, outbreaks of LPDV have been reported from Europe and Israel. Although the virus has previously never been detected in North America, herein we describe the widespread distribution, genetic diversity, pathogenesis, and evolution of LPDV in the United States. Characterization of the provirus genome of the index LPDV case from North America demonstrated an 88% nucleotide identity to the Israeli prototype strain. Although phylogenetic analysis indicated that the majority of viruses fell into a single North American lineage, a small subset of viruses from South Carolina were most closely related to the Israeli prototype. These results suggest that LPDV was transferred between continents to initiate outbreaks of disease. However, the direction (New World to Old World or vice versa), mechanism, and time frame of the transcontinental spread currently remain unknown.

Peninsular bighorn sheep (Ovis canadensis nelsoni) are found exclusively in Southern California and Baja Mexico. They are federally endangered due to multiple threats, including introduced infectious disease. From 1981 - 2017, we conducted surveillance for 16 pathogens and estimated population sizes, adult survival, and lamb survival. We used mixed effects regression models to assess disease patterns at the individual and population levels. Pathogen infection/exposure prevalence varied both spatially and temporally. Our findings indicate that the primary predictor of individual pathogen infection/exposure was the region in which an animal was captured, implying that transmission is driven by local ecological or behavioral factors. Higher Mycoplasma ovipneumoniae seropositivity was associated with lower lamb survival, consistent with lambs having high rates of pneumonia-associated mortality, which may be slowing population recovery. There was no association between M. ovipneumoniae and adult survival. Adult survival was positively associated with population size and parainfluenza-3 virus seroprevalence in the same year, and orf virus seroprevalence in the previous year. Peninsular bighorn sheep are recovering from small population sizes in a habitat of environmental extremes, compounded by infectious disease. Our research can help inform future pathogen surveillance and population monitoring for the long-term conservation of this population.

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Since 1998, cyclic mortality events in common eiders (Somateria mollissima), numbering in the hundreds to thousands of dead birds, have been documented along the coast of Cape Cod, MA, USA. Although longitudinal disease investigations have uncovered potential contributing factors responsible for these outbreaks, detecting a primary etiological agent has proven enigmatic. Here, we identify a novel orthomyxovirus, tentatively named Wellfleet Bay virus (WFBV), as a potential causative agent of these outbreaks. Genomic analysis of WFBV revealed that it is most closely related to members of the Quaranjavirus genus within the family Orthomyxoviridae. Similar to other members of the genus, WFBV contains an alphabaculovirus gp64-like glycoprotein that was demonstrated to have fusion activity; this also tentatively suggests that ticks (and/or insects) may vector the virus in nature. However, in addition to the six RNA segments encoding the prototypical structural proteins identified in other quaranjaviruses, a previously unknown RNA segment (segment 7) encoding a novel protein designated VP7 was discovered in WFBV. Although WFBV shows low to moderate levels of sequence similarity to Quaranfil virus and Johnston Atoll virus, the original members of the Quaranjavirus genus, additional antigenic and genetic analyses demonstrated that it is closely related to the recently identified Cygnet River virus (CyRV) from South Australia, suggesting that WFBV and CyRV may be geographic variants of the same virus. Although the identification of WFBV in part may resolve the enigma of these mass mortality events, the details of the ecology and epidemiology of the virus remain to be determined.

Importance

The emergence or reemergence of viral pathogens resulting in large-scale outbreaks of disease in humans and/or animals is one of the most important challenges facing biomedicine. For example, understanding how orthomyxoviruses such as novel influenza A virus reassortants and/or mutants emerge to cause epidemic or pandemic disease is at the forefront of current global health concerns. Here, we describe the emergence of a novel orthomyxovirus, Wellfleet Bay virus (WFBV), which has been associated with cyclic large-scale bird die-offs in the northeastern United States. This initial characterization study provides a foundation for further research into the evolution, epidemiology, and ecology of newly emerging orthomyxoviruses, such as WFBV, and their potential impacts on animal and/or human health.