Canine Central Nervous System Neoplasm Phenotyping Using Tissue Microarray Technique.

Tissue microarrays (TMAs) represent a useful technique for the simultaneous phenotyping of large sample numbers and are particularly suitable for histopathologic tumor research. In this study, TMAs were used to evaluate semiquantitatively the expression of multiple antigens in various canine central nervous system (CNS) neoplasms and to identify markers with potential discriminative diagnostic relevance. Ninety-seven canine CNS neoplasms, previously diagnosed on hematoxylin and eosin sections according to the World Health Organization classification, were investigated on TMAs, with each tumor consisting of 2 cylindrical samples from the center and the periphery of the neoplasm. Tumor cells were phenotyped using a panel of 28 monoclonal and polyclonal antibodies, and hierarchical clustering analysis was applied to group neoplasms according to similarities in their expression profiles. Hierarchical clustering generally grouped cases with similar histologic diagnoses; however, gliomas especially exhibited a considerable heterogeneity in their positivity scores. Multiple tumor groups, such as astrocytomas and oligodendrogliomas, significantly differed in the proportion of positive immunoreaction for certain markers such as p75NTR, AQP4, GFAP, and S100 protein. The study highlights AQP4 and p75NTR as novel markers, helping to discriminate between canine astrocytoma and oligodendroglioma. Furthermore, the results suggest that p75NTR and proteolipid protein may represent useful markers, whose expression inversely correlates with malignant transformation in canine astrocytomas and oligodendrogliomas, respectively. Tissue microarray was demonstrated to be a useful and time-saving tool for the simultaneous immunohistochemical characterization of multiple canine CNS neoplasms. The present study provides a detailed overview of the expression patterns of different types of canine CNS neoplasms.

Polycystic kidneys and GM2 gangliosidosis-like disease in neonatal springboks (Antidorcas marsupialis).

Clinical, gross, histopathologic, electron microscopic findings and enzymatic analysis of 4 captive, juvenile springboks (Antidorcas marsupialis) showing both polycystic kidneys and a storage disease are described. Springbok offspring (4 of 34; 12%) were affected by either one or both disorders in a German zoo within a period of 5 years (2008-2013). Macroscopic findings included bilaterally severely enlarged kidneys displaying numerous cysts in 4 animals and superior brachygnathism in 2 animals. Histopathologically, kidneys of 4 animals displayed cystic dilation of the renal tubules. In addition, abundant cytoplasmic vacuoles with a diameter ranging from 2 to 10 µm in neurons of the central and peripheral nervous system, hepatocytes, thyroid follicular epithelial cells, pancreatic islets of Langerhans and renal tubular cells were found in 2 springbok neonates indicative of an additional storage disease. Ultrastructurally, round electron-lucent vacuoles, up to 4 µm in diameter, were present in neurons. Enzymatic analysis of liver and kidney tissue of 1 affected springbok revealed a reduced activity of total hexosaminidase (Hex) with relatively increased HexA activity at the same level of total Hex, suggesting a hexosaminidase defect. Pedigree analysis suggested a monogenic autosomal recessive inheritance for both diseases. In summary, related springboks showed 2 different changes resembling both polycystic kidney and a GM2 gangliosidosis similar to the human Sandhoff disease. Whether the simultaneous occurrence of these 2 entities represents an incidental finding or has a genetic link needs to be investigated in future studies.

Salient lesions in domestic ruminants infected with the emerging so-called Schmallenberg virus in Germany.

The so-called Schmallenberg virus (SBV), first detected in a German town of the same name in October 2011, is a novel emerging orthobunyavirus in Europe causing malformations and severe economic loss in ruminants. This report describes lesions in 40 sheep, 2 goats, and 16 cattle naturally infected with SBV as determined by real-time quantitative reverse transcription polymerase chain reaction. The most common macroscopic changes were arthrogryposis, vertebral malformations, brachygnathia inferior, and malformations of the central nervous system, including hydranencephaly, porencephaly, hydrocephalus, cerebellar hypoplasia, and micromyelia. Histologic lesions included lymphohistiocytic meningoencephalomyelitis in some cases, glial nodules mainly in the mesencephalon and hippocampus of lambs and goats, and neuronal degeneration and necrosis mainly in the brain stem of calves. Micromyelia was characterized by a loss of gray and white matter, with few neurons remaining in the ventral horn in calves. The skeletal muscles had myofibrillar hypoplasia in lambs and calves. The lesions of SBV-associated abortion and perinatal death are similar to those attributed to Akabane virus and other viruses in the Simbu group of bunyaviruses.

Neurotoxic potential of reactive astrocytes in canine distemper demyelinating leukoencephalitis.

Canine distemper virus (CDV) causes a fatal demyelinating leukoencephalitis in young dogs resembling human multiple sclerosis. Astrocytes are the main cellular target of CDV and undergo reactive changes already in pre-demyelinating brain lesions. Based on their broad range of beneficial and detrimental effects in the injured brain reactive astrogliosis is in need of intensive investigation. The aim of the study was to characterize astrocyte plasticity during the course of CDV-induced demyelinating leukoencephalitis by the aid of immunohistochemistry, immunofluorescence and gene expression analysis. Immunohistochemistry revealed the presence of reactive glial fibrillary acidic protein (GFAP)+ astrocytes with increased survivin and reduced aquaporin 4, and glutamine synthetase protein levels, indicating disturbed blood brain barrier function, glutamate homeostasis and astrocyte maladaptation, respectively. Gene expression analysis revealed 81 differentially expressed astrocyte-related genes with a dominance of genes associated with neurotoxic A1-polarized astrocytes. Accordingly, acyl-coA synthetase long-chain family member 5+/GFAP+, and serglycin+/GFAP+ cells, characteristic of A1-astrocytes, were found in demyelinating lesions by immunofluorescence. In addition, gene expression revealed a dysregulation of astrocytic function including disturbed glutamate homeostasis and altered immune function. Observed findings indicate an astrocyte polarization towards a neurotoxic phenotype likely contributing to lesion initiation and progression in canine distemper leukoencephalitis.

Fatal Disseminated Toxoplasma gondii Infection in a Captive Harbour Porpoise (Phocoena phocoena).

A 7-year-old female harbour porpoise (Phocoena phocoena), born and held in captivity, suffered from reduced consciousness, imprecise and circling swimming movements and long phases of immobility over a period of 3 weeks. The animal died during treatment in a Danish open sea facility. Pathological examination revealed multifocal pyogranulomatous to necrotizing meningoencephalomyelitis, ganglioneuritis, plexus chorioiditis, myocarditis, hepatitis and adrenalitis with few intralesional protozoal tachyzoites and bradyzoites within cysts. Immunohistochemistry was positive for Toxoplasma gondii antigen within the lesions. Using polymerase chain reaction (PCR), the presence of T. gondii-specific genome fragments was confirmed. A multilocus PCR-restriction fragment length polymorphism analysis using nine unlinked marker regions (nSAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1 and Apico) resulted in the identification of T. gondii type II (variant Apico Type I), which is the T. gondii genotype dominating in Germany. This is the first description of disseminated fatal toxoplasmosis in a captive harbour porpoise that lived in an open sea basin. Surface water contaminated with toxoplasma oocysts is regarded as the most likely source of infection.

Lack of schmallenberg virus in ruminant brain tissues archived from 1961 to 2010 in Germany.

Schmallenberg virus (SBV) is an orthobunyavirus of the family Bunyaviridae that is associated with stillbirth and malformations in ruminants. The infection has been identified in many European countries since August 2011. The present study investigated retrospectively the occurrence of SBV infection in ruminants using immunohistochemistry and in-situ hybridization in brain tissues archived between 1961 and 2010 (112 cattle, 57 sheep, 16 goats and 27 wild ruminants). Eighty-five animals with inflammatory brain lesions and 47 animals with malformations were included. Due to the lack of SBV protein and RNA detection, SBV appears to have been introduced recently into Northern parts of Europe from tropical or subtropical regions.

Skeletal muscle hypoplasia represents the only significant lesion in peripheral organs of ruminants infected with Schmallenberg virus during gestation.

Schmallenberg virus (SBV), an arbovirus within the family Bunyaviridae, represents a ruminant pathogen that has caused epidemic abortion and birth of malformed or stillborn animals in many European countries since August 2011. Histological and immunohistochemical analysis of peripheral tissues of SBV-infected animals, including lymphoid tissues, endocrine organs and tissues of the gastrointestinal, urogenital and respiratory system, were analyzed in order to elucidate the occurrence of SBV-associated changes and the presence of viral antigens and RNA. Twenty calves and 12 lambs as well as age-matched controls were included in this study. Significant muscular hypoplasia with fatty replacement was noted in affected calves and lambs. In addition, hepatocellular degeneration with lymphohistiocytic inflammation, interstitial fibrosis and biliary hyperplasia was detected in calves. All animals lacked SBV-positive cells in the peripheral organs. These observations resemble those found in Akabane virus- and Cache Valley virus-infected animals and support the occurrence of few residual lesions in peripheral organs following SBV infection.

Equid herpesvirus 5-associated dermatitis in a horse--Resembling herpes-associated erythema multiforme.

An equid herpesvirus 5 (EHV-5) infection was detected in lesioned skin from a nine-year-old Holsteiner stallion in the south of Germany. Macroscopically, the animal displayed a non-pruritic, multifocal, pustular dermatitis around both eyes, nostrils and the muzzle, which had been ongoing for one year. Histopathologically, skin lesions were characterized by orthokeratotic to parakeratotic hyperkeratosis, pustular dermatitis, epidermal hyperplasia, apoptotic keratinocytes, a lympho-plasmahistiocytic interface dermatitis with hydropic degeneration of keratinocytes, and perivascular to diffuse, lympho-histiocytic infiltrations. The stratum granulosum and the upper part of the stratum spinosum contained multiple amphophilic, intranuclear inclusion bodies. By in situ hybridization and immunohistochemistry herpesvirus DNA and protein, respectively, were detected within keratinocytes containing inclusion bodies. Sequencing of the PCR-product revealed the presence of EHV-5 DNA. This is the first description of a dermatitis associated with EHV-5 in a horse, indicating that EHV-5 should be considered as an etiology of lymphohistiocytic interface dermatitis with intranuclear inclusion bodies in horses and is similar to herpes-associated erythema multiforme in humans.

Spinal epidermoid cyst in a SJL mouse: case report and literature review.

This report is the first description of a spinal epidermoid cyst (EC) in a SJL mouse and gives an overview on the occurrence of ECs in animals including dogs, horses, mice and rats. The EC was not detected grossly and the mouse did not display clinical signs or an altered rotarod performance. Microscopically, there was an oval cyst lined by stratified squamous epithelium that was attached to the dorsolateral meninges and caused moderate compression of the adjacent lumbar spinal cord. ECs in mice and rats are mainly located in the caudal part of the spinal cord with a variable, strain-dependent occurrence. ECs in mice and rats are not associated with clinical signs and can be interpreted as incidental findings.