Arno Wünschmann

Pathologic Findings in Red-Tailed Hawks (Buteo jamaicensis) and Cooper's Hawks (Accipiter cooperi) Naturally Infected with West Nile Virus

Abstract Carcasses of 13 red-tailed hawks (RTHAs) and 11 Coopers hawks (COHAs) were tested for West Nile virus (WNV) using WNV-specific reverse transcriptase–polymerase chain reaction (RT-PCR) on fresh brain tissue and WNV-specific immunohistochemistry (IHC) on various organs. Ten COHAs (91%) and 11 RTHAs (85%) were positive for WNV RNA by RT-PCR. All 11 COHAs (100%) and 10 RTHAs (77%) were positive for WNV antigen by IHC. A triad of inflammatory lesions, including chronic lymphoplasmacytic and histiocytic encephalitis, endophthalmitis, and myocarditis, was common in both species. In COHAs, the heart (54%), cerebrum (50%), and eye (45%) were the organs that most commonly contained WNV antigen. The amount of WNV antigen was usually small. In RTHAs, the kidney (38%), cerebrum (38%), cerebellum (38%), and eye (36%) were the organs most commonly containing WNV antigen. Unlike COHAs, larger amounts of WNV antigen were present in the cerebrum of RTHAs. WNV antigen was detected in similar cell populations in both species, including neurons of brain, spinal cord, and retina, pigmented epithelial cells of the retina, epithelial cells of renal medullary tubules, cardiomyocytes, endothelial cells and smooth muscle cells of arteries, dendritic cells of splenic lymph follicles, exocrine pancreatic cells, adrenal cells, and keratinocytes of the skin. The study presents strong evidence that WNV can cause a chronic fatal disease in RTHAs and COHAs. The lesion distribution of WNV infection in both species is variable, but inflammatory lesions are common, and a triad of lesions including encephalitis, myocarditis, and endophthalmitis is indicative of WNV infection in both species.

Pathologic and Immunohistochemical Findings in Goshawks (Accipiter gentilis) and Great Horned Owls (Bubo virginianus) Naturally Infected with West Nile Virus

Abstract The carcasses of 25 great horned owls and 12 goshawks were investigated for West Nile virus (WNV) infection by immunohistochemistry (IHC) performed on various organs, including brain, spinal cord, heart, kidney, eye, bone marrow, spleen, liver, lungs, pancreas, intestine, and proventriculus, using a WNV-antigen–specific monoclonal antibody and by WNV-specific reverse transcriptase-polymerase chain reaction (RT-PCR), performed on fresh brain tissue only. WNV infection was diagnosed by IHC in all owls and all goshawks. WNV-specific RT-PCR amplified WNV-RNA in the brain of all goshawks but only 12 owls (48%). Cachexia was a common macroscopic finding associated with WNV infection in owls (76%). Myocarditis was occasionally macroscopically evident in goshawks (33%). Microscopically, inflammatory lesions, including lymphoplasmacytic and histiocytic encephalitis, myocarditis, endophthalmitis, and pancreatitis were present in both species but were more common and more severe in goshawks than in owls. The most characteristic brain lesion in owls was the formation of glial nodules, in particular in the molecular layer of the cerebellum, while encephalitis affecting the periventricular parenchyma of the cerebral cortex was common in the goshawks. In owls, WNV-antigen–positive cells were present usually only in very small numbers per organ. Kidney (80%), heart (39%), and cerebellum (37%) were the organs that most commonly contained WNV antigen in owls. WNV antigen was frequently widely distributed in the organs of infected goshawks, with increased amounts of WNV antigen in the heart and the cerebrum. Spleen (75%), cerebellum (66%), heart (58%), cerebrum (58%), and eye (50%) were often WNV-antigen positive in goshawks. In contrast with the goshawks, WNV antigen was not present in cerebral and retinal neurons of owls. WNV infection appears to be capable of causing fatal disease in great horned owls and goshawks. However, the distribution and severity of histologic lesions, the antigen distribution in the various organs, and the amount of antigen varied among both species. Therefore, the diagnostician may choose organs for histology and immunohistochemistry as well as RT-PCR depending on the investigated species in order to avoid false-negative results.

Caprine Herpesvirus-2–Associated Malignant Catarrhal Fever in White-Tailed Deer (Odocoileus Virginianus)

A subacute disease presenting primarily as alopecia and weight loss occurred in 2 white-tailed deer (Odocoileus virginianus) on farms in Minnesota and in Texas. A presumptive diagnosis of malignant catarrhal fever (MCF) was made on the basis of histological lesions. Antibody against an epitope conserved among the MCF group viruses was detected in the serum of both deer. DNA samples from the deer were subjected to a variety of PCR amplifications. Alignment of the amplified sequences from the diseased animals revealed that they were 100% identical to each other and to the same DNA fragment from the newly recognized member of the MCF virus group endemic in domestic goats (Capra hircus), provisionally named caprine herpesvirus 2 (CpHV-2). A seroprevalence survey from one of the deer farms showed a high rate of subclincal infection in the deer population. This study provides further confirmation that CpHV-2 is a pathogen, at least for deer, and emphasizes the risk of loss from MCF when mixing cervids with goats.

Pathological and Immunohistochemical Findings in American Crows (Corvus Brachyrhynchos) Naturally Infected with West Nile Virus

Twenty-one American crows were identified as being West Nile virus (WNV) infected by WNV-specific reverse transcriptase-polymerase chain reaction (RT-PCR) performed on fresh brain tissue (cerebrum and cerebellum of 16 crows) or by WNV-specific immunohistochemistry of various organs (21 crows). Consistent gross lesions attributable to WNV infection were not detected. Common histological lesions included necrosis of spleen and bone marrow. West Nile virus antigen was consistently detected in heart and kidney (100%). In addition, bone marrow (92%), duodenum (89%), proventriculus (87%), liver (86%), lung (85%), spleen (80%), pancreas (61%), and brain (45%) contained WNV antigen-positive cells. Infected cells included cardiomyocytes; neurons; endothelial cells and vascular smooth muscle cells; hematopoietic cells of bone marrow; and macrophages of spleen, liver (Kupffer cells), and lungs. Epithelial cells of renal tubules, duodenum, pancreas, and proventriculus were also infected. The diagnostic histopathologist should consider WNV infection in crows in the absence of any inflammatory lesions. Immunohistochemistry of heart and kidney is as reliable in detecting WNV infection in American crows as RT-PCR of fresh brain tissue.

Cerebral cysticercosis by Taenia crassiceps in a domestic cat

Cerebral cysticercosis by Taenia crassiceps was diagnosed in an adult female domestic shorthair cat. The animal was euthanized 6 weeks after the initial presentation with signs of vomiting, lethargy, and ataxia. The disease took an intermittent relapsing course with the neurological signs progressing eventually to recumbancy and coma. At necropsy, numerous cysticerci were found in the dilated left lateral ventricle and the adjacent brain parenchyma. The cysticerci were identified as metacestodes of T. crassiceps larvae based on size and morphology of the cysts; shape, number, and size of the rostellar hooks; and mode of proliferation, including endogenous and exogenous budding. Cerebral cysticercosis by T. crassiceps is rare in atypical intermediate hosts and has not been described in cats.

Sarcocystis sp.-Associated Meningoencephalitis in a Bald Eagle (Haliaeetus Leucocephalus)

Protozoal meningoencephalitis is uncommon in raptors. An adult female bald eagle (Haliaeetus leucocephalus) was euthanized after several months of treatment for progressive neurologic signs. The predominant histologic lesion was lymphoplasmacytic and histiocytic meningoencephalitis involving the cerebrum and cerebellum. There was a marked segmental loss of granular cells and Purkinje cells, as well as segmental atrophy of the molecular layer in the cerebellum. Protozoal merozoites and schizonts were observed in the gray matter of the cerebellum. Ultrastructurally, the merozoites were classified as a species of Sarcocystis due to the lack of rhoptries. Immunohistochemistry of the agent revealed a positive reaction for Sarcocystis neurona, while sections were negative for Toxoplasma gondii and Neospora caninum. Sarcocystis sp. infection should be considered as a differential diagnosis in bald eagles with chronic neurologic disease.

Sarcocystis calchasi-Associated Neurologic Disease in a Domestic Pigeon in North America

Tissue cysts of a protozoan parasite were present in the skeletal muscle of a domestic pigeon (Columba livia f. domestica) with neurologic disease in Minnesota, USA. The animal had a severe granulomatous meningoencephalitis. The cysts were slender, up to 1 mm long and up to 0.03 mm in diameter. The cysts had a smooth wall without projections. Size and wall morphology were compatible with Sarcocystis calchasi. Polymerase chain reaction using S. calchasi-specific primers resulted in a specific amplicon from the skeletal muscle but not from the brain. Sequencing of the highly variable genomic regions ITS1 and D2 revealed 100% nucleic acid identity with the German strain of S. calchasi. Sarcocystis calchasi is the cause of an emerging lethal disease in pigeons in Germany. This is the first description of the parasite outside of Germany.

Sarcocystis falcatula-associated encephalitis in a free-ranging great horned owl (Bubo virginianus).

A great horned owl (Bubo virginianus) was admitted to a rehabilitation clinic with severe neurologic signs that were unresponsive to supportive care. The animal was euthanatized because of a poor prognosis. Marked granulomatous encephalitis with focal brainstem malacia was detected microscopically. The brainstem was the most severely affected brain location and the only place in which schizonts and merozoites, morphologically compatible with Sarcocystis spp., were detected. Immunohistochemistry with the use of polyclonal antisera indicated the presence of Sarcocystis falcatula. The species identification of the protozoa as S. falcatula was confirmed by polymerase chain reaction. To the authors knowledge, this is the first report of spontaneous S. falcatula–associated encephalitis in a great horned owl.

Clinical Findings, Lesions, and Viral Antigen Distribution in Great Gray Owls (Strix nebulosa) and Barred Owls (Strix varia) with Spontaneous West Nile Virus Infection

Abstract West Nile Virus (WNV) infection manifests itself clinically and pathologically differently in various species of birds. The clinicopathologic findings and WNV antigen tissue distribution of six great gray owls (Strix nebulosa) and two barred owls (Strix varia) with WNV infection are described in this report. Great gray owls usually live in northern Canada, whereas the phylogenetically related barred owls are native to the midwestern and eastern United States and southern Canada. Naturally acquired WNV infection caused death essentially without previous signs of disease in the six great gray owls during a mortality event. Lesions of WNV infection were dominated by hepatic and splenic necrosis, with evidence of disseminated intravascular coagulation in the great gray owls. WNV antigen was widely distributed in the organs of the great gray owls and appeared to target endothelial cells, macrophages, and hepatocytes. The barred owls represented two sporadic cases. They had neurologic disease with mental dullness that led to euthanasia. These birds had mild to moderate lymphoplasmacytic encephalitis with glial nodules and lymphoplasmacytic pectenitis. WNV antigen was sparse in barred owls and only present in a few brain neurons and renal tubular epithelial cells. The cause of the different manifestations of WNV disease in these fairly closely related owl species is uncertain.

Enteric Coronavirus Infection in a Juvenile Dromedary (Camelus Dromedarius)

A case of an enteric coronavirus infection in a 6-week-old dromedary calf is described. The animal had diarrhea for 5 days and died despite symptomatic treatment. Numerous viral particles, approximately 140 nm in diameter, with club-like projections were detected in the feces by electron microscopy. These characteristics were consistent with a coronavirus. Immunohistochemical reactivity with 2 antigenic group II coronavirus-specific antibodies confirmed the presence of viral antigen in colonic epithelial cells. The death of the animal was attributed to a neutrophilic and emphysematous colitis that likely was caused by an infection with a Clostridium sp.