SARS-CoV-2 exposure in wild white-tailed deer (Odocoileus virginianus).

Widespread human SARS-CoV-2 infections combined with human-wildlife interactions create the potential for reverse zoonosis from humans to wildlife. We targeted white-tailed deer (Odocoileus virginianus) for serosurveillance based on evidence these deer have angiotensin-converting enzyme 2 receptors with high affinity for SARS-CoV-2, are permissive to infection, exhibit sustained viral shedding, can transmit to conspecifics, exhibit social behavior, and can be abundant near urban centers. We evaluated 624 prepandemic and postpandemic serum samples from wild deer from four US states for SARS-CoV-2 exposure. Antibodies were detected in 152 samples (40%) from 2021 using a surrogate virus neutralization test. A subset of samples tested with a SARS-CoV-2 virus neutralization test showed high concordance between tests. These data suggest white-tailed deer in the populations assessed have been exposed to SARS-CoV-2.

Bourbon Virus in Wild and Domestic Animals, Missouri, USA, 2012-2013.

Since its recent discovery, Bourbon virus has been isolated from a human and ticks. To assess exposure of potential vertebrate reservoirs, we assayed banked serum and plasma samples from wildlife and domestic animals in Missouri, USA, for Bourbon virus-neutralizing antibodies. We detected high seroprevalence in raccoons (50%) and white-tailed deer (86%).

A Bead-Based Flow Cytometric Assay for Monitoring Yersinia pestis Exposure in Wildlife.

Yersinia pestis is the causative agent of plague and is considered a category A priority pathogen due to its potential for high transmissibility and the significant morbidity and mortality it causes in humans. Y. pestis is endemic to the western United States and much of the world, necessitating programs to monitor for this pathogen on the landscape. Elevated human risk of plague infection has been spatially correlated with spikes in seropositive wildlife numbers, particularly rodent-eating carnivores, which are frequently in contact with the enzootic hosts and the associated arthropod vectors of Y. pestis In this study, we describe a semiautomated bead-based flow cytometric assay developed for plague monitoring in wildlife called the F1 Luminex plague assay (F1-LPA). Based upon Luminex/Bio-Plex technology, the F1-LPA targets serological responses to the F1 capsular antigen of Y. pestis and was optimized to analyze antibodies eluted from wildlife blood samples preserved on Nobuto filter paper strips. In comparative evaluations with passive hemagglutination, the gold standard tool for wildlife plague serodiagnosis, the F1-LPA demonstrated as much as 64× improvement in analytical sensitivity for F1-specific IgG detection and allowed for unambiguous classification of IgG status. The functionality of the F1-LPA was demonstrated for coyotes and other canids, which are the primary sentinels in wildlife plague monitoring, as well as felids and raccoons. Additionally, assay formats that do not require species-specific immunological reagents, which are not routinely available for several wildlife species used in plague monitoring, were determined to be functional in the F1-LPA.

Pseudorabies detected in hunting dogs in Alabama and Arkansas after close contact with feral swine (Sus scrofa).

BACKGROUND: Pigs (Sus scrofa) are the natural hosts of pseudorabies virus (PRV), also known as Aujeszky's disease. Infection in mammals, with the exception of humans, typically causes extreme itching, facial swelling, and excessive salivation, followed by death in non-suid species. The risk to susceptible mammals was assumed to decrease when PRV was eliminated from U.S. commercial swine in 2004, though the virus remains endemic in feral swine. Infected feral swine pose a threat to the disease-free status of the commercial swine industry, and to other animals, including dogs, that come in direct or indirect contact with them. Since dogs are commonly used for hunting feral swine, they are at high risk of exposure. CASE PRESENTATION: The following report describes the progression of pseudorabies infection in dogs in two states after exposure to feral swine. The first case occurred in a dog in Alabama after participation in a competitive wild hog rodeo. The second case occurred in multiple dogs in Arkansas after hunting feral swine, and subsequent consumption of the offal. The antibody prevalence of feral swine in the two states where the dogs were exposed is also examined. CONCLUSIONS: Dogs that are used for hunting feral swine are at high risk of exposure to pseudorabies because the disease is considered endemic in feral swine in the U.S.

Feral swine virome is dominated by single-stranded DNA viruses and contains a novel Orthopneumovirus which circulates both in feral and domestic swine.

Feral swine are known reservoirs for various pathogens that can adversely affect domestic animals. To assess the viral ecology of feral swine in the USA, metagenomic sequencing was performed on 100 pooled nasal swabs. The virome was dominated by small, ssDNA viruses belonging to the families Circoviridae, Anelloviridae and Parvovirinae. Only four RNA viruses were identified: porcine kobuvirus, porcine sapelovirus, atypical porcine pestivirus and a novel Orthopneumovirus, provisionally named swine orthopneumovirus (SOV). SOV shared ~90 % nucleotide identity to murine pneumonia virus (MPV) and canine pneumovirus. A modified, commercially available ELISA for MPV found that approximately 30 % of both feral and domestic swine sera were positive for antibodies cross-reactive with MPV. Quantitative reverse transcription-PCR identified two (2 %) and four (5.0 %) positive nasal swab pools from feral and domestic swine, respectively, confirming that SOV circulates in both herds.

Novel Eurasian highly pathogenic avian influenza A H5 viruses in wild birds, Washington, USA, 2014.

Novel Eurasian lineage avian influenza A(H5N8) virus has spread rapidly and globally since January 2014. In December 2014, H5N8 and reassortant H5N2 viruses were detected in wild birds in Washington, USA, and subsequently in backyard birds. When they infect commercial poultry, these highly pathogenic viruses pose substantial trade issues.

Disease Progression and Serological Assay Performance in Heritage Breed Pigs following Brucella suis Experimental Challenge as a Model for Naturally Infected Feral Swine.

Invasive feral swine (Sus scrofa) are one of the most important wildlife species for disease surveillance in the United States, serving as a reservoir for various diseases of concern for the health of humans and domestic animals. Brucella suis, the causative agent of swine brucellosis, is one such pathogen carried and transmitted by feral swine. Serology assays are the preferred field diagnostic for B. suis infection, as whole blood can be readily collected and antibodies are highly stable. However, serological assays frequently have lower sensitivity and specificity, and few studies have validated serological assays for B. suis in feral swine. We conducted an experimental infection of Ossabaw Island Hogs (a breed re-domesticated from feral animals) as a disease-free proxy for feral swine to (1) improve understanding of bacterial dissemination and antibody response following B. suis infection and (2) evaluate potential changes in the performance of serological diagnostic assays over the course of infection. Animals were inoculated with B. suis and serially euthanized across a 16-week period, with samples collected at the time of euthanasia. The 8% card agglutination test performed best, whereas the fluorescence polarization assay demonstrated no capacity to differentiate true positive from true negative animals. From a disease surveillance perspective, using the 8% card agglutination test in parallel with either the buffered acidified plate antigen test or the Brucella abortus/suis complement fixation test provided the best performance with the highest probability of a positive assay result. Application of these combinations of diagnostic assays for B. suis surveillance among feral swine would improve understanding of spillover risks at the national level.

Adaptive risk-based targeted surveillance for foreign animal diseases at the wildlife-livestock interface.

Animal disease surveillance is an important component of the national veterinary infrastructure to protect animal agriculture and facilitates identification of foreign animal disease (FAD) introduction. Once introduced, pathogens shared among domestic and wild animals are especially challenging to manage due to the complex ecology of spillover and spillback. Thus, early identification of FAD in wildlife is critical to minimize outbreak severity and potential impacts on animal agriculture as well as potential impacts on wildlife and biodiversity. As a result, national surveillance and monitoring programs that include wildlife are becoming increasingly common. Designing surveillance systems in wildlife or, more importantly, at the interface of wildlife and domestic animals, is especially challenging because of the frequent lack of ecological and epidemiological data for wildlife species and technical challenges associated with a lack of non-invasive methodologies. To meet the increasing need for targeted FAD surveillance and to address gaps in existing wildlife surveillance systems, we developed an adaptive risk-based targeted surveillance approach that accounts for risks in source and recipient host populations. The approach is flexible, accounts for changing disease risks through time, can be scaled from local to national extents and permits the inclusion of quantitative data or when information is limited to expert opinion. We apply this adaptive risk-based surveillance framework to prioritize areas for surveillance in wild pigs in the United States with the objective of early detection of three diseases: classical swine fever, African swine fever and foot-and-mouth disease. We discuss our surveillance framework, its application to wild pigs and discuss the utility of this framework for surveillance of other host species and diseases.

Influenza A virus surveillance, infection and antibody persistence in snow geese (Anser caerulescens).

Some snow geese (Anser caerulescens) migrate between Eurasia and North America and exhibit high seroprevalence for influenza A viruses (IAVs). Hence, these birds might be expected to play a role in intercontinental dispersal of IAVs. Our objective in this manuscript was to characterize basic incidence and infection characteristics for snow geese to assess whether these birds are likely to significantly contribute to circulation of IAVs. Thus, we 1) estimated snow goose infection prevalence by summarizing > 5,000 snow goose surveillance records, 2) experimentally infected snow geese with a low pathogenic IAV (H4N6) to assess susceptibility and infection dynamics and 3) characterized long-term antibody kinetics. Infection prevalence based on surveillance data for snow geese was 7.88%, higher than the infection rates found in other common North American goose species. In the experimental infection study, only 4 of 7 snow geese shed viral RNA. Shedding in infected birds peaked at moderate levels (mean peak 102.62 EID50 equivalents/mL) and was exclusively associated with the oral cavity. Serological testing across a year post-exposure showed all inoculated birds seroconverted regardless of detectable shedding. Antibody levels peaked at 10 days post-exposure and then waned to undetectable levels by 6 months. In sum, while broad-scale surveillance results showed comparatively high infection prevalence, the experimental infection study showed only moderate susceptibility and shedding. Consequently, additional work is needed to assess whether snow geese might exhibit higher levels of susceptibility and shedding rates when exposed to other IAV strains.

Comparative susceptibility of eastern cottontails and New Zealand white rabbits to classical rabbit haemorrhagic disease virus (RHDV) and RHDV2.

Rabbit haemorrhagic disease virus (RHDV) is associated with high morbidity and mortality in the European rabbit (Oryctolagus cuniculus). In 2010, a genetically distinct RHDV named RHDV2 emerged in Europe and spread to many other regions, including North America in 2016. Prior to this study it was unknown if eastern cottontails (ECT(s); Sylvilagus floridanus), one of the most common wild lagomorphs in the United States, were susceptible to RHDV2. In this study, 10 wild-caught ECTs and 10 New Zealand white rabbits (NZWR(s); O. cuniculus) were each inoculated orally with either RHDV (RHDVa/GI.1a; n = 5 per species) or RHDV2 (a recombinant GI.1bP-GI.2; n = 5 per species) and monitored for the development of disease. Three of the five ECTs that were infected with RHDV2 developed disease consistent with RHD and died at 4 and 6 days post-inoculation (DPI). The RHDV major capsid protein/antigen (VP60) was detected in the livers of three ECTs infected with RHDV2, but none was detected in the ECTs infected with RHDV. Additionally, RHD viral RNA was detected in the liver, spleen, intestine and blood of ECTs infected with RHDV2, but not in the ECTs infected with RHDV. RHD viral RNA was detected in urine, oral swabs and rectal swabs in at least two of five ECTs infected with RHDV2. One ECT inoculated with RHDV2 seroconverted and developed a high antibody titre by the end of the experimental period (21 DPI). ECTs inoculated with the classic RHDV did not seroconvert. In comparison, NZWRs inoculated with RHDV2 exhibited high mortality (five of five) at 2 DPI and four of five NZWRs inoculated with RHDV either died or were euthanized at 2 DPI indicating both of these viruses were highly pathogenic to this species. This experiment indicates that ECTs are susceptible to RHDV2 and can shed viral RNA, thereby suggesting this species could be involved in the epidemiology of this virus.

Epizootic hemorrhagic disease outbreak in a captive facility housing white-tailed deer (Odocoileus virginianus), bison (Bison bison), elk (Cervus elaphus), cattle (Bos taurus), and goats (Capra hircus) in Colorado, U.S.A.

An ungulate research facility in Fort Collins, Colorado, U.S.A., experienced mortality in white-tailed deer (Odocoileus virginianus) because of epizootic hemorrhagic disease virus (EHDV) infection from 20 August 2007 through 26 September 2007. Epizootic hemorrhagic disease virus (EHDV) was detected by reverse transcriptase polymerase chain reaction and virus isolation from the spleen and lung tissues of two white-tailed deer. Virus neutralization tests were performed on pre- and postoutbreak sera from other species maintained in the same facility, including bison (Bison bison), elk (Cervus elaphus), domestic cattle (Bos taurus), and domestic goats (Capra hircus), as well as postoutbreak sera from the surviving white-tailed deer. Serum samples that represented all species in the facility neutralized EHDV-1 and EHDV-2 either before or after the outbreak. The animals that neutralized EHDV-1 did not neutralize EHDV-2. No clinical signs attributable to EHDV infection were noted in any of the species other than the deer during the outbreak. Although experimental EHDV infections have been reported in bison and elk, natural exposures have not been previously documented in these species in North America. The roles that elk, bison, cattle, and goats might play in the epidemiology of EHDV in a close-contact multispecies situation remain unknown.

FOOT-AND-MOUTH DISEASE IN EXPERIMENTALLY INFECTED MULE DEER (ODOCOILEUS HEMIONUS).

The only known outbreak of foot-and-mouth disease (FMD) in wildlife in the US occurred in mule deer (Odocoileus hemionus) in California in 1924-25. There is little recorded information on the pathogenesis and epidemiology of the disease in deer in that outbreak. In this experimental study, we compared the susceptibility of mule deer to FMD virus (FMDV) serotype O to that of cattle (Bos taurus). We also determined the potential for intra- and interspecies transmission of FMDV serotype O in mule deer and cattle, and assessed conventional laboratory tests in their ability to detect FMDV in mule deer. Two mule deer and one steer were each infected by intraepithelial tongue inoculation with 10,000 bovine tongue infective doses of FMDV, strain O1 Manisa. The inoculated steer and deer were kept in the same room with contact animals of both species. Exposed contact animals were moved to rooms with unexposed animals after becoming febrile. All mule deer (n=14) and cattle (n=6) developed clinical signs and lesions consistent with FMDV infection. Deer had a high prevalence of myocarditis and high mortality. Virus was transmitted between mule deer, from cattle to mule deer, and from mule deer to cattle. Virus and antibodies against nonstructural FMDV proteins in mule deer and cattle were detected by conventional laboratory tests. Virus shedding was detected by PCR and virus isolation up to 9 d postexposure in deer.

Antemortem detection of PrPCWD in preclinical, ranch-raised Rocky Mountain elk (Cervus elaphus nelsoni) by biopsy of the rectal mucosa.

Antemortem biopsy of the rectal mucosa was evaluated as a method for the preclinical diagnosis of chronic wasting disease (CWD) in a herd of ranch-raised Rocky Mountain elk (Cervus elaphus nelsoni) quarantined because of exposure to CWD. Biopsy samples were obtained from 41 elk during the winter of 2005-2006 and from 26 elk from that herd still alive and available for testing during the winter of 2006-2007. Samples were examined for PrP(CWD), the protein marker for CWD infection, by immunohistochemistry. PrP(CWD) was detected in follicles of the rectoanal mucosa-associated lymphoid tissue in biopsy samples from 1 elk with clinical signs of chronic wasting disease and 5 clinically normal elk. The diagnosis was confirmed in all 6 animals by postmortem analysis of brain and peripheral lymph nodes. PrP(CWD) was also observed in the submucosal plexus and myenteric plexus of the enteric nervous system, and in close association with nonmyelinated mucosal and submucosal nerve fibers. In antemortem rectal biopsy samples from positive animals, immunostaining was consistently observed in approximately 60% of the mucosa-associated lymphoid tissue follicles if 10 or more total follicles per biopsy were present for evaluation. Most antemortem biopsy samples obtained from elk younger than 6.5 years contained at least 10 follicles per rectal mucosal biopsy. These findings support the analysis of antemortem biopsy of the rectal mucosa samples as part of an integrated strategy to manage chronic wasting disease in Rocky Mountain elk.

Current status and future recommendations for feral swine disease surveillance in the United States.

USDA APHIS Wildlife Services (WS) responded to the threat of feral swine as a pathogen reservoir as early as 2004. To increase awareness and knowledge on that risk, WS began opportunistic sampling of animals harvested by its operational component to curtail swine damage to agriculture and property. Initially, pseudorabies and swine brucellosis were of most concern, as both serve as a potential threat to the domestic swine industry and the latter also possesses zoonotic implications. In 2006, classical swine fever, a foreign animal disease, became the main driver for feral swine pathogen surveillance. Subsequent years of surveillance identified numerous other disease risks inherent within populations of feral swine. Presently, feral swine surveillance falls under the purview of the APHIS National Feral Swine Damage Management Program, which began in 2014. In January 2018, a panel of animal disease experts representing industry, government, and academia were invited to Fort Collins, Colorado to discuss successes of this surveillance, identify any shortcomings or needs, and propose future feral swine surveillance. This manuscript serves to synthesize WS' surveillance and the future direction of these efforts.

Surveillance of coyotes to detect bovine tuberculosis, Michigan.

Bovine tuberculosis (TB) is endemic in white-tailed deer (Odocoileus virginianus) in the northeastern portion of Michigan's Lower Peninsula. Bovine TB in deer and cattle has created immense financial consequences for the livestock industry and hunting public. Surveillance identified coyotes (Canis latrans) as potential bio-accumulators of Mycobacterium bovis, a finding that generated interest in their potential to serve as sentinels for monitoring disease risk. We sampled 175 coyotes in the bovine TB-endemic area. Fifty-eight tested positive, and infection prevalence by county ranged from 19% to 52% (statistical mean 33%, SE 0.07). By contrast, prevalence in deer (n = 3,817) was lower (i.e., 1.49%; Mann-Whitney U4,4 = 14, p<0.001). By focusing on coyotes rather than deer, we sampled 97% fewer individuals and increased the likelihood of detecting M. bovis by 40%. As a result of reduced sampling intensity, sentinel coyote surveys have the potential to be practical indicators of M. bovis presence in wildlife and livestock.

Chronic wasting disease in a Wisconsin white-tailed deer farm.

In September 2002, chronic wasting disease (CWD), a prion disorder of captive and wild cervids, was diagnosed in a white-tailed deer (Odocoileus virginianus) from a captive farm in Wisconsin. The facility was subsequently quarantined, and in January 2006 the remaining 76 deer were depopulated. Sixty animals (79%) were found to be positive by immunohistochemical staining for the abnormal prion protein (PrP(CWD)) in at least one tissue; the prevalence of positive staining was high even in young deer. Although none of the deer displayed clinical signs suggestive of CWD at depopulation, 49 deer had considerable accumulation of the abnormal prion in the medulla at the level of the obex. Extraneural accumulation of the abnormal protein was observed in 59 deer, with accumulation in the retropharyngeal lymph node in 58 of 59 (98%), in the tonsil in 56 of 59 (95%), and in the rectal mucosal lymphoid tissue in 48 of 58 (83%). The retina was positive in 4 deer, all with marked accumulation of prion in the obex. One deer was considered positive for PrP(CWD) in the brain but not in the extraneural tissue, a novel observation in white-tailed deer. The infection rate in captive deer was 20-fold higher than in wild deer. Although weakly related to infection rates in extraneural tissues, prion genotype was strongly linked to progression of prion accumulation in the obex. Antemortem testing by biopsy of recto-anal mucosal-associated lymphoid tissue (or other peripheral lymphoid tissue) may be a useful adjunct to tonsil biopsy for surveillance in captive herds at risk for CWD infection.

Foot-and-mouth disease in North American bison (Bison bison) and elk (Cervus elaphus nelsoni): susceptibility, intra- and interspecies transmission, clinical signs, and lesions.

There is limited information about the pathogenesis and epidemiology of foot-and-mouth disease (FMD) in North American bison (Bison bison) or elk (Cervus elaphus nelsoni). In these two experimental infection studies, we compared the susceptibilities of bison and elk to FMD virus (FMDV), respectively, with that of cattle; determined whether intra- and interspecies transmission could occur in bison and cattle, and elk and cattle; determined suitability of conventional available laboratory tests to detect FMDV infection in bison and elk; and investigated whether bison or elk are efficient long-term carriers of FMDV. In both studies, after a period of acclimation to the containment at Plum Island Animal Disease Center, animals were infected by intraepithelial tongue inoculation with 10,000 bovine tongue infective doses of FMDV, strain O1 Manisa. Inoculated animals were kept with contact animals; subsequently, inoculated and/or exposed contact animals were placed in rooms with unexposed animals. All bison developed oral mucosal and foot lesions similar to those of cattle. Bison developed fever, lameness, inappetence, and ptyalism. Physical examinations on bison revealed numerous small vesicles and erosions affecting tongue, gingiva, muzzle, hard and soft palates, coronary bands, and interdigital skin. Inoculated elk developed transient fever and mild focal tongue and foot lesions. Contact elk developed neither clinical signs nor gross pathologic lesions of FMD. At necropsy, lesions in bison included numerous extensive vesicles, erosions, and/or ulcers in the oral cavities, feet, and rumen pillars depending on the stage of disease. Less extensive oral, foot, and rumen lesions were present in the inoculated elk. All bison and inoculated elk developed antibodies to FMDV and were positive for FMDV by reverse transcription-polymerase chain reaction (RT-PCR). Transmission occurred between cattle and bison, and bison and bison. It did not occur between elk and cattle. Elk-to-elk transmission studies resulted in only one contact elk developing serologic evidence of a subclinical infection. Other exposed elk developed neither clinical, pathologic, virologic, nor serologic evidence of disease. FMDV was not isolated from animals past 28 days postinfection.

LEPTOSPIRA ANTIBODIES DETECTED IN WILDLIFE IN THE USA AND THE US VIRGIN ISLANDS.

From 2011 to 2017, 4,534 serum samples from 13 wildlife species collected across the US and in one territory (US Virgin Islands) were tested for exposure to Leptospira serovars Bratislava, Canicola, Grippotyphosa, Hardjo, Icterohaemorrhagiae, and Pomona. Of 1,759 canids, 1,043 cervids, 23 small Indian mongooses ( Herpestes auropunctatus), 1,704 raccoons ( Procyon lotor), and five striped skunks ( Mephitis mephitis), 27.0, 44.4, 30.4, 40.8, and 60%, respectively, were antibody positive for any of the six serovars. The most commonly detected serovars across all species were Bratislava and Grippotyphosa. Our results indicate that Leptospira titers are very common in a wide variety of wildlife species. These species may act as important reservoirs in the epidemiological cycle of the pathogen. Additional studies to determine the relationship between serologic evidence and shedding of the pathogen by wildlife are necessary to better understand the risk.

Elk with a long incubation prion disease phenotype have a unique PrPd profile.

The transmissible spongiform encephalopathies (TSEs) invariably result in fatal neurodegeneration and accumulation of PrP, an abnormal form of the host prion protein PrP, encoded by the PRNP gene. A naturally occurring polymorphism (methionine/valine) at PRNP codon 129 is associated with variation in relative disease susceptibility, incubation time, clinical presentation, neuropathology, and/or PrP biochemical characteristics in a range of human TSEs. A methionine/leucine polymorphism at the corresponding site in the Rocky Mountain elk PRNP gene is associated with variation in relative susceptibility and incubation time in the cervid TSE chronic wasting disease. We now report that elk lacking the predisposing 132-methionine allele develop chronic wasting disease after a long incubation period and display a novel PrP folding pattern.

Antibodies to Various Zoonotic Pathogens Detected in Feral Swine (Sus scrofa) at Abattoirs in Texas, USA.

The zoonotic risk posed to employees by slaughtering feral swine (Sus scrofa) at two abattoirs in Texas was assessed by testing feral swine serum samples for exposure to influenza A virus, Leptospira, Trichinella spiralis, and Toxoplasma gondii. Blood was collected from a total of 376 feral swine between the two facilities during six separate collection periods in 2015. Antibodies to one or more serovars of Leptospira were identified in 48.9% of feral swine tested, with Bratislava and Pomona as the most commonly detected serovars, and antibodies to influenza A virus were detected in 14.1% of feral swine. Antibodies to T. gondii and T. spiralis were identified in 9.0 and 3.5%, respectively, of feral swine tested. Our results suggest that abattoir employees should be aware of the potential for exposure to various zoonotic pathogens when slaughtering feral swine, wear appropriate personal protective equipment, and participate in medical monitoring programs to ensure detection and prompt treatment. In addition, consumers of feral swine should cook the meat to the appropriate temperature and wash hands and kitchen surfaces thoroughly after preparing meat.