SURVEILLANCE FOR AN EMERGENT HOOF DISEASE IN ELK (CERVUS ELAPHUS) IN THE US PACIFIC WEST SUPPLEMENTED BY 16S RRNA GENE AMPLICON SEQUENCING.

A novel hoof disease of elk (Cervus elaphus) was described in southwestern Washington, US, in 2008 and was subsequently diagnosed in an adjacent area in northwestern Oregon in 2014. The disease, currently referred to as treponeme-associated hoof disease (TAHD), is characterized by lesions ranging from mild erosions, to severe ulcers with underrunning of the hoof capsule and heel-sole junction, to overgrown and avulsed hoof capsules. Histologically, lesions exhibit epithelial erosion or ulceration, suppurative inflammation, and the presence of argyrophilic spirochetes. We used data collected by the Washington Department of Fish and Wildlife and Oregon Department of Fish and Wildlife from 2008 to 2017 as reference for disease distribution. We then conducted enhanced surveillance in 2018-20 by obtaining 164 submissions from four US Pacific West states. We detected TAHD for the first time in Idaho and northern California, as well as in multiple counties in Washington and Oregon where it had not been previously reported. Given the unexpectedly broad disease distribution, continued surveillance is warranted to determine the full geographic extent of TAHD. From samples of 22 elk, we investigated 16S rRNA gene amplicon sequencing as a technique that could be used to supplement TAHD surveillance. Operational taxonomic units of the family Spirochaetaceae were identified in 10 of 12 histologically diagnosed TAHD-positive cases and two of 10 TAHD-negative cases. Phyla Spirochaetae (P<0.008), Fusobacteria (P<0.006), and Tenericutes (P<0.01) were overrepresented in samples from TAHD-positive feet when compared with TAHD-negative elk. A unique spirochete, PT19, was detected in hooves of 11 elk and from at least one elk in each state. Results support the use of 16S rRNA gene amplicon sequencing as a reliable and informative tool to supplement investigations into distribution and etiology of this presumed polybacterial disease.

Computed tomographic diagnosis of necroulcerative reticulorumenitis with portal venous gas in a lamb.

A 2-month-old, intact male Merino sheep was presented for acute inappetence and pyrexia. Contrast-enhanced abdominal computed tomography identified reticulorumenal intramural gas with suspected ruminal ulceration, arborizing portal venous gas, and mild peritoneal fluid. The lamb was treated medically for presumptive ulcerative reticulorumenitis and non-septic peritonitis. Over 10 days, the lamb initially showed mild improvement before clinical deterioration and was ultimately euthanized. Definitive diagnosis of necrotizing, ulcerative reticulorumenitis was made via necropsy with histopathology. An underlying cause was not determined. Key clinical message: Contrast-enhanced computed tomography was effective in identifying gastric emphysema and portal venous gas associated with reticuloruminal ulceration.

Diagnostic par tomodensitométrie d'une réticulo-ruménite nécro-ulcérative avec gaz veineux porte chez un agneau. Un mouton mérinos mâle intact, âgé de 2 mois, a été présenté pour une inappétence aiguë et une pyrexie. La tomodensitométrie abdominale de contraste augmentée a identifié du gaz intramural réticuloruménal avec une ulcération ruminale suspectée, du gaz veineux porte arborescent et un léger liquide péritonéal. L'agneau a été traité médicalement pour une réticulo-ruménite ulcéreuse présumée et une péritonite non-septique. Sur une période de 10 jours, l'agneau a d'abord montré une légère amélioration avant une détérioration clinique et a finalement été euthanasié. Le diagnostic définitif de réticulo-ruménite ulcéreuse nécrosante a été posé suite à la nécropsie et l'histopathologie. Une cause sous-jacente n'a pas été déterminée.Message clinique clé:La tomodensitométrie assistée par contraste s'est avérée efficace pour identifier l'emphysème gastrique et le gaz veineux porte associés à l'ulcération réticulo-ruminale.(Traduit par Dr Serge Messier).

Viruses in unexplained encephalitis cases in American black bears (Ursus americanus).

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.

Comparison of multilocus sequence types found among North American isolates of Mycoplasma bovis from cattle, bison, and deer, 2007-2017.

A prior multilocus sequence typing (MLST) study reported that Mycoplasma bovis isolates from North American bison possess sequence types (STs) different from those found among cattle. The 42 bison isolates evaluated were obtained in 2007 or later, whereas only 19 of 94 (~20%) of the available cattle isolates, with only 1 from North America, were from that same time. We compared STs of additional, contemporary, North American cattle isolates with those from bison, as well as isolates from 2 North American deer, all originating during the same timeframe, to more definitively assess potential strain-related host specificity and expand our understanding of the genetic diversity of M. bovis. From 307 isolates obtained between 2007 and 2017 (209 from cattle, 96 from bison, 2 from deer), we identified 49 STs, with 39 found exclusively in cattle and 5 exclusively in bison. Four STs were shared between bison and cattle isolates; one ST was found in cattle and in a deer. There was no clear association between ST and the health status of the animal of origin. An MLST-based phylogeny including 41 novel STs identified in our study reveals that STs found in bison fall within several divergent lineages that include STs found exclusively in cattle.

Anabaena/Dolichospermum as the source of lethal microcystin levels responsible for a large cattle toxicosis event.

Thirty-two 14-month old steers died during a period of four days (19-23 June 2017) after drinking from Junipers Reservoir (southeastern Oregon, USA) during a cyanobacterial bloom. Clinical and histopathological findings were consistent with acute liver disease, and microcystin-LR was present at 3000 µg/L in a reservoir water sample and at 7100 µg/L in the rumen contents of one of the mortalities. Serum biochemistry and histological examination indicated severe liver damage consistent with microcystin toxicosis. Microscopic observation of reservoir water samples, limited to frozen or poorly stored and partially degraded samples, indicated the presence of abundant Anabaena/Dolichospermum, but the presence of other toxic cyanobacteria such as Microcystis could not be excluded. Metagenomic analysis showed the presence in these samples of a single cyanobacterium whose cpcBA, rpoB and rbcL genes indicated membership in the Anabaena/Dolichospermum genus. The sequence of a complete mcy gene cluster with homology to previously identified Anabaena mcy genes was recovered. These results emphasize the capacity for Anabaena/Dolichospermum blooms to produce lethal levels of microcystin, posing a danger to public health and livestock. Further, our findings indicate that such occurrences can occur outside the far-northern latitudes in which microcystin-producing Anabaena have typically been found.

Viral testing of 10 cases of Theiler's disease and 37 in-contact horses in the absence of equine biologic product administration: A prospective study (2014-2018).

BACKGROUND: A novel equine parvovirus (EqPV-H) was recently discovered in the equine liver with Theiler's disease. OBJECTIVES: To determine the prevalence of EqPV-H infection in naturally occurring Theiler's disease cases and in-contact horses in the absence of historical equine biologic product administration. ANIMALS: Ten cases of Theiler's disease from 6 separate properties were included in the study, based on the criteria of acute onset of clinical signs of liver failure with laboratory or histopathologic findings characteristic of Theiler's disease and no history of receiving an equine biologic product within the preceding 4 months. In addition, 37 in-contact horses from 4 of the 6 properties were screened for EqPV-H infection and hepatitis. METHODS: In prospective case series, cases were diagnosed with Theiler's disease by the attending veterinarian and were tested for EqPV-H by PCR of liver or serum. In-contact horses were assessed via serum chemistry and PCR at the attending veterinarian's discretion. Hepatitis was defined as serum gamma-glutamyltransferase activity above reference interval. The association of EqPV-H with hepatitis was determined by Fisher's exact test. RESULTS: Nine of 10 (90%) Theiler's disease cases and 54% of tested in-contact horses were EqPV-H positive. Hepatitis was significantly associated with EqPV-H infection (P = .036). CONCLUSIONS AND CLINICAL IMPORTANCE: Although further study is required to identify EqPV-H as the causative agent of Theiler's disease, EqPV-H appears strongly associated with cases of fatal Theiler's disease and subclinical hepatitis in horses in contact with those cases. The prevalence of EqPV-H infection on affected properties can be high.

Aspergillus Section Nigri-Associated Calcium Oxalate Crystals in an Eurasian Eagle Owl (Bubo bubo).

An adult male Eurasian eagle owl (Bubo bubo) housed at a wildlife rehabilitation facility in southern Oregon died after a short period of progressive ill-thrift. Radiographs taken prior to death demonstrated abnormal radiopaque material in the coelom and the owl was submitted for postmortem examination. Black pigmented fungus was noted grossly, particularly in the respiratory tissues, with abundant oxalate crystal deposition associated with and without hyphal elements subsequently observed histologically. Aspergillus section Nigri was cultured from the lesions. Although there have been a few reports of aspergillosis caused by Aspergillus niger in avian species, the severity and wide tissue distribution of oxalates in this case are highly unusual.

High copy number of ovine gammaherpesvirus 2 DNA associated with malignant catarrhal fever-like syndrome in a lamb.

Domestic and wild sheep are the natural reservoirs for ovine gammaherpesvirus 2 (OvHV-2), the causative agent of sheep-associated malignant catarrhal fever (SA-MCF). Virtually all adult sheep are infected with OvHV-2 under natural flock conditions, and infection is normally subclinical. MCF-like clinical signs and typical histologic lesions in sheep have been linked during case investigations at veterinary diagnostic laboratories; however, the confirmation of naturally occurring MCF in sheep is problematic. To date, the assays for detection of OvHV-2-specific antibodies or DNA are usually positive in sheep, regardless of health status, so mere detection of antibodies or the agent is of minimal diagnostic significance in this species. We document herein a naturally occurring MCF case in a 4-mo-old domestic lamb and demonstrate that the affected animal had 100-1,000 times more OvHV-2 copy numbers in tissues than in healthy adult and age-matched sheep. These results indicate that high copy numbers of viral DNA in tissues associated with characteristic lesions can be used to confirm the diagnosis of MCF in sheep.

Alcelaphine gammaherpesvirus 1-induced malignant catarrhal fever in a Watusi ( Bos taurus africanus) steer in a North American game park.

A 10-y-old Watusi ( Bos taurus africanus) steer housed at a drive-through game park in Winston, Oregon developed severe clinical illness including fever, marked nasal discharge, injected scleral and conjunctival membranes, plus oral hemorrhages and erosions. The animal responded poorly to supportive treatment and was euthanized. Additional gross findings at postmortem examination included papules and erosive lesions on the tongue, hemorrhagic large intestine, and multifocal cardiac hemorrhages. Histopathologic findings included multifocal lymphoplasmacytic vasculitis plus fibrin exudation in heart and tongue. Total DNA obtained from the splenic samples was positive for alcelaphine gammaherpesvirus 1 (AlHV-1) as tested by a multiplex PCR for malignant catarrhal fever (MCF) viruses. The AlHV-1 detection was further confirmed by amplification and sequencing of a viral DNA polymerase gene fragment, which was identical to AlHV-1 sequences in GenBank. This was the first diagnosis of clinical wildebeest-associated MCF on these premises, although wildebeest have been held at the park for over 25 y. This disease is sporadic in North America and should be considered as a differential diagnosis for febrile illness with ulcerative oral lesions in ruminants.

Disease Surveillance of California Ground Squirrels ( Spermophilus beecheyi ) in a Drive-through Zoo in Oregon, USA.

Rodents and other small wild mammals are often considered to be pests and vectors for disease in zoos that house small populations of valuable threatened and endangered animals. In 2005, three nonhuman primates at a drive-through zoo in Oregon, US, acquired tularemia from an unknown source. Due to an abundance of California ground squirrels ( Spermophilus beecheyi ) on zoo grounds, we instituted serosurveillance of this species from July through September 2008 to determine the prevalence of antibodies against pathogens considered to be potentially transmissible to collection animals. Serologic testing was performed for Francisella tularensis ; Leptospira interrogans serovars Canicola, Grippotyphosa, Hardjo, Icterohemorrhagiae, and Pomona; Toxoplasma gondii ; and Yersinia pestis . All squirrels were seronegative for Yersinia pestis (0%; 0/45) and Toxoplasma gondii (0%; 0/20); there was a prevalence of 2% (1/45) for Francisella tularensis antibodies and 57% (24/42) were positive for various Leptospira serovars. Although it remains unclear whether ground squirrels present a significant risk for transmission of disease to zoo animals, vaccination of high-risk zoo animals against leptospirosis warrants consideration. Beyond this, continued vigilance and persistence with various forms of pest control may reduce the likelihood of disease transmission from wildlife hosts to animals in human care.

Rhabdochlamydia spp. in an Oregon raptor.

PCR-based approach was used to examine the rate of Chlamydia positivity in raptors from wild bird rehabilitation centers in Oregon. Three of 82 birds were identified as positive for Chlamydia with this PCR. Sequence analysis of 16S ribosomal DNA from 2 of these birds confirmed the presence of DNA from phylum Chlamydiae. One bird was positive for Chlamydia psittaci in both choanal and cloacal swabs. The second bird, a louse-infested red-tailed hawk, had evidence of choanal colonization by "Candidatus Rhabdochlamydia" spp. Our study describes evidence of this Chlamydia-like organism in the United States. This survey also suggests that the carriage rate of C. psittaci is low in raptors in Oregon wild bird rehabilitation centers, and that care must be taken in the design of PCR primers for phylum Chlamydiae such that colonization by insect endosymbionts is not mistaken for an infection by known chlamydial pathogens.

A comparison of microscopic ink characteristics of 35 commercially available surgical margin inks.

OBJECTIVE: To compare microscopic characteristics of commercially available surgical margin inks used for surgical pathology specimens. STUDY DESIGN: Prospective in vitro study. SAMPLE POPULATION: Thirty-five different surgical margin inks (black, blue, green, orange, red, violet, and yellow from 5 different manufacturers). METHODS: Inks were applied to uniform, single-source, canine cadaveric full-thickness ventral abdominal tissue blocks. Tissue blocks and ink manufacturers were randomly paired and each color was applied to a length of the cut tissue margin. After drying, tissues were fixed in formalin, and 3 radial slices were obtained from each color section and processed for routine histologic evaluation, yielding 105 randomly numbered slides with each manufacturer's color represented in triplicate. Slides were evaluated by 5 blinded, board-certified veterinary anatomic pathologists using a standardized scoring scheme. Statistical analyses were performed to evaluate for ink manufacturer effects on scores, correlation among different subjective variables, and pathologist agreement. RESULTS: Black and blue had the most consistently high scores whereas red and violet had the most consistently low overall scores, across all manufacturers. All colors tested, except yellow, had statistically significant differences in overall scores among individual manufacturers. Overall score was significantly correlated to all other subjective microscopic scores evaluated. The average Spearman correlation coefficient among the 10 pairwise pathologists overall ink scores was 0.60. CONCLUSIONS: There are statistically significant differences in microscopic ink characteristics among manufacturers, with a notable degree of inter-pathologist agreement.

Interaction of Mycobacterium leprae with human airway epithelial cells: adherence, entry, survival, and identification of potential adhesins by surface proteome analysis.

This study examined the in vitro interaction between Mycobacterium leprae, the causative agent of leprosy, and human alveolar and nasal epithelial cells, demonstrating that M. leprae can enter both cell types and that both are capable of sustaining bacterial survival. Moreover, delivery of M. leprae to the nasal septum of mice resulted in macrophage and epithelial cell infection in the lung tissue, sustaining the idea that the airways constitute an important M. leprae entry route into the human body. Since critical aspects in understanding the mechanisms of infection are the identification and characterization of the adhesins involved in pathogen-host cell interaction, the nude mouse-derived M. leprae cell surface-exposed proteome was studied to uncover potentially relevant adhesin candidates. A total of 279 cell surface-exposed proteins were identified based on selective biotinylation, streptavidin-affinity purification, and shotgun mass spectrometry; 11 of those proteins have been previously described as potential adhesins. In vitro assays with the recombinant forms of the histone-like protein (Hlp) and the heparin-binding hemagglutinin (HBHA), considered to be major mycobacterial adhesins, confirmed their capacity to promote bacterial attachment to epithelial cells. Taking our data together, they suggest that the airway epithelium may act as a reservoir and/or portal of entry for M. leprae in humans. Moreover, our report sheds light on the potentially critical adhesins involved in M. leprae-epithelial cell interaction that may be useful in designing more effective tools for leprosy control.

Diagnostic sampling and gross pathology of New World camelids.

This article provides an overview of tests and appropriate samples to send to a Veterinary Diagnostic Laboratory for the diagnosis of common diseases of New World Camelids (NWC) such as abortions, congenital anomalies, anemia, enteritis, endoparasitism, gastric ulcer, hepatic lipidosis, encephalitis, pneumonia, dermatosis, neoplasia and cryptococcosis. Unique anatomic features of NWC and common findings encountered during gross necropsy examination are briefly reviewed.

Fatal Caprine arthritis encephalitis virus-like infection in 4 Rocky Mountain goats (Oreamnos americanus).

Over a 3.5-year period, 4 Rocky Mountain goats (Oreamnos americanus), housed at a single facility, developed clinical disease attributed to infection by Caprine arthritis encephalitis virus (CAEV). Ages ranged from 1 to 10 years. Three of the goats, a 1-year-old female, a 2-year-old male, and a 5-year-old male, had been fed raw domestic goat milk from a single source that was later found to have CAEV on the premises. The fourth animal, a 10-year-old male, had not ingested domestic goat milk but had been housed with the other 3 Rocky Mountain goats. All 4 animals had clinical signs of pneumonia prior to death. At necropsy, findings in lungs included marked diffuse interstitial pneumonia characterized histologically by severe lymphoplasmacytic infiltrates with massive alveolar proteinosis, interstitial fibrosis, and type II pneumocyte hyperplasia. One animal also developed left-sided hemiparesis, and locally extensive lymphoplasmacytic myeloencephalitis was present in the cranial cervical spinal cord. Two animals had joint effusions, as well as severe lymphoplasmacytic and ulcerative synovitis. Immunohistochemical staining of fixed sections of lung tissue from all 4 goats, as well as spinal cord in 1 affected animal, and synovium from 2 affected animals were positive for CAEV antigen. Serology testing for anti-CAEV antibodies was positive in the 2 goats tested. The cases suggest that Rocky Mountain goats are susceptible to naturally occurring CAEV infection, that CAEV from domestic goats can be transmitted to this species through infected milk and by horizontal transmission, and that viral infection can result in clinically severe multisystemic disease.

Alopecia areata in two black Angus cows.

Adult onset progressive alopecia and leukotrichia developed in 2 registered black Angus cows, aged 6 and 7 years. Histopathologic findings in skin were prominent melanin clumping and degeneration of matrix cells with formation of giant multinucleate cells within hair bulbs, accompanied by peribulbar melanin incontinence and fibrosis and dystrophic hair shafts. Intrabulbar and peribulbar lymphocytes were cluster of differentiation (CD)3-positive T cells. Findings were characteristic of alopecia areata.

Thiafentanil-dexmedetomidine-telazol anesthesia in greater rheas (Rhea americana).

Ratite anesthetic events are often dangerous because these birds use their powerful legs and clawed feet as a defense, and physical restraint can result in self-trauma or injury to handlers. Although various combinations of opioids, alpha-2 adrenergic agonists, and dissociative agents have been employed in ratites, few effective chemical immobilization protocols have been documented for rheas (Rhea spp.). An intramuscular, remote-delivered combination of thiafentanil (0.30 +/- 0.08 mg/kg), dexmedetomidine (7.31 +/- 2.72 microg/kg), and tiletamine-zolazepam (5.09 +/- 2.31 mg/kg) was utilized in eight adult (four male, four female) greater rheas (Rhea americana). Smooth inductions were observed. During clinical procedures, birds were intubated and maintained on isoflurane gas, and atipamezole was administered to antagonize the dexmedetomidine. At recovery, naltrexone was administered to antagonize the thiafentanil, and midazolam was administered to smooth crate recoveries until release. This low-volume, high-potency, reversible drug combination demonstrated safe inductions and smooth recoveries and proved to be a reliable anesthetic regimen for greater rheas.

Cylicospirura species (Nematoda: Spirocercidae) and stomach nodules in cougars (Puma concolor) and bobcats (Lynx rufus) in Oregon.

The stomachs and proximal duodena of 160 cougars (Puma concolor) and 17 bobcats (Lynx rufus), obtained throughout Oregon during 7 yr, were examined for Cylicospirura spp. and associated lesions. Prevalence in cougars was 73%, with a range in intensity of 1-562 worms. The mean diameter of nodules was 1.2 cm (SD=0.5), and many extended through the submucosa to the muscularis. About 83% of cougars had nodules; most nodules contained worms, but 14% of the smaller nodules (<0.2 cm) contained porcupine (Erethizon dorsatum) quills. A mean of 12.4 worms/nodule (SD=34.1) was observed, with a maximum of 340 worms/nodule. Prevalence in bobcats was 53%, with an intensity of 1-25 worms. About 65% of bobcats had nodules, which were slightly smaller than those in cougars but appeared to involve similar layers of gastrointestinal tissue. One to 25 Cylicospirura sp. were found in all but two small nodules in bobcats. Cougars killed for livestock damage or safety concerns had a significantly higher median worm intensity than did those that died of other causes. Also, the median worm intensity of older cougars was higher than that of younger lions. There were more males than females killed for livestock damage or safety concerns. The cylicospirurid from cougars was Cylicospirura subaequalis, and that of bobcats was Cylicospirura felineus. These two similar species were separated morphologically by differences in tooth and sex organ morphology. They were also differentiated by DNA sequence analysis of the mitochondrial cytochrome c oxidase subunit 1 gene (cox1). Worm sequences from cougars differed from those from bobcats by 11%, whereas essentially no difference was found among worms from the same host. Phylogenetic analysis showed that within the order Spirurida, both cylicospirurids were most closely related to Spirocerca lupi, based on this gene sequence.