A STORAGE-LIKE DISEASE WITH SKELETAL INVOLVEMENT IN A DOG

A storage-like disease in a 14-month-old Rhodesian Ridgeback dog is described. Clinically the dog was lame on all legs, and appendicular joint capsules were distended. Radiographically and at necropsy, there was evidence of osteoarthritis, increased volume of synovial fluid, and marked hypertrophy of the synovial membranes. Large mononclear cells with vacuolated cytoplasm were evident in synovial membranes. Large mononuclear cells with vacuolated cytoplasm were evident in synovial membranes. Large mononuclear cells with vacuolated cytoplasm were evident in synovial membranes, bone marrow, liver, lymph nodes, and endocrine glands. Vacuolated cells in lymph nodes stained positively for fat. Transmission electron microscopy revealed empty membrane-bound cytoplasmic vacuoles. The similarity of this disease to storage diseases in man where arthropathy is a feature is discussed.     

Lipid-rich pleural mesothelioma in a dog

An 11-year-old, neutered, male Golden Retriever cross dog was euthanized following a history of recurrent pericardial effusions. At necropsy, blood-tinged pericardial and intrathoracic effusions were seen along with numerous firm to hard plaque-like masses that studded the epicardial, pericardial, mediastinal, and costal pleural surfaces. Within the right thorax, the lesions coalesced into a large mass that occupied most of the cavity. Histologically, the masses were composed of solid sheets and papillary aggregates of medium-sized polygonal cells that contained abundant vacuolated to clear cytoplasm. Some of the cytoplasmic vacuoles stained positive with oil red O. The stroma contained metaplastic trabeculae of woven and lamellar bone. Immunohistochemically, the neoplastic cells expressed vimentin, pancytokeratin, and S-100 protein. Transmission electron microscopy corroborated the presence of intracytoplasmic vacuoles and demonstrated prominent intercellular junctional complexes and apically located microvilli. These findings are consistent with a lipid-rich variant of mesothelioma. To the authors' knowledge, this is the first report of a lipid-rich mesothelioma in a dog.     

A Case of Spontaneous Malignant Hibernoma in a Crl:CD(SD)IGS Rat

A firm, tan, well-circumscribed mass that measured 25 × 30 × 35 mm was observed in the thoracic cavity of a 53-week-old male Crl:CD(SD) IGS rat. Histologically, the mass was encapsulated by fibrous tissue and contained fibrovascular septae. Tumor cells were compactly arranged, and most were oval to polygonal in shape with multivacuolated cytoplasm and a centrally located nucleus. In some parts of the tumor, marked cellular atypia and frequent mitoses were evident. Vacuoles in cytoplasm were positive for oil red O. The tumor cells were characterized ultrastructurally by abundant, round to oval mitochondria with transverse closely-packed cristae. Tumor cells were immunohistochemically positive for uncoupling protein 1 (UCP-1). Several thrombi and hemorrhagic or necrotic foci were also observed within the tumor mass. Vascular invasion of the tumor capsule was observed; however, invasion of surrounding tissues or metastases were not observed. Based on the pathology findings, this case was diagnosed as a malignant hibernoma.     

Epididymal interstitial (Leydig) cell tumors in B6C3F1 mice

Six primary interstitial cell tumors of the epididymis were identified from 46,752 male B6C3F1 mice used in chronic toxicity and carcinogenicity studies. Five of the tumors occurred at the end of 2-year studies; none were attributed to treatment. None of the mice with epididymal tumors had a primary testicular tumor. Histologically, tumors were characterized by a nodular or diffuse proliferation of tumor cells in the epididymal interstitium. Most cells were polygonal with highly vacuolated cytoplasm (vacuolated cells) or eosinophilic cytoplasm (eosinophilic cells). Smaller hyperchromatic cells with scant basophilic cytoplasm (basophilic cells) and cells with yellow-brown pigment characteristic of lipofuscin (pigmented cells) were less common. In each tumor two or more cell types were present. Extension of these tumors through the capsule, invasion of the testis, or metastasis did not occur. By electron microscopy both eosinophilic and vacuolated cell types had a large round or oval nucleus with sparse heterochromatin, abundant mitochondria with tubulovesicular cristae, and frequent desmosome structures between cell membranes. Vacuolated cells contained numerous lipid droplets. Morphological features of the epididymal tumors are similar to those of the testicular interstitial (Leydig) cell tumor in mice and rats.     

A spontaneous thymic carcinosarcoma in a young Sprague Dawley rat

We encountered a case of spontaneous thymic carcinosarcoma in a young Crl:CD (Sprague Dawley) rat. Grossly, a white multinodular mass replaced the thymus in the thoracic cavity. Histologically, multiple nodules were separated by fibrous stroma, and each nodule included isolated regions that were composed of epithelial or non-epithelial tumor cells. The epithelial tumor cells were relatively large and round to polygonal cells with large nuclei and weakly eosinophilic cytoplasm. These cells were cytokeratin-positive and vimentin-negative. These cells infiltrated the lungs. The non-epithelial tumor cells were poorly differentiated, small, round to spindle-shaped cells with small nuclei and basophilic cytoplasm. These cells were vimentin-positive and mostly cytokeratin-negative. Many islands of cartilage were observed near non-epithelial cells. Based on these findings, the tumor was diagnosed as a primary thymic carcinosarcoma consisting of a malignant thymoma composed of epithelial tumor cells and a mesenchymal chondrosarcoma composed of non-epithelial tumor cells.     

Mixed germ cell tumor in the eye of a dog.

A 3-year-old female neutered Staffordshire Bull Terrier presented with a mixed germ cell tumor involving the base of the iris and the ciliary body of the right eye. The tumor mass was composed primarily of packeted vacuolated, polygonal (hepatoid) cells and small round cells; epithelial cells lining tubuloacinar structures were a less prominent component. The hepatoid and round cells stained positively for alpha-fetoprotein and cytokeratin. The epithelial cells stained positively for cytokeratin only, and some contained cytoplasmic mucin droplets. The polygonal cells were interpreted as a hepatoid variant of yolk sac tumor, and the epithelial cells were considered a teratomatous component. Trabeculae of bone were observed within the mass and may have been metaplastic or a teratomatous element. Extragonadal germ cell tumors are rare in dogs and have previously been reported only in the suprasellar region. This is the first report of this tumor type in the eye of a nonhuman species.     

Coccygeal chordoma in a dog.

A 4-cm diameter spherical mass was removed from the region of the second coccygeal vertebra of a 6-year-old female Doberman Pinscher dog. The mass had been present for 7 months and was described as multilobular and smooth. Histologically, the neoplasm consisted of small spindle-shaped cells surrounding lobules of large vacuolated polygonal cells. Mucin was present within most lobules. The large polygonal cells stained positively with antibodies against vimentin, cytokeratin, and S-100 protein. Although the spindle-shaped cells stained positively with antibodies against vimentin, they only occasionally contained cytokeratin. The histology and immunohistochemistry are consistent with that reported for chordoma. Although 4 chordomas have previously been reported in dogs, this is the first case in which immunohistochemistry has been used to support the histological diagnosis. Neither recurrence nor metastasis was reported 10 months after surgical excision.     

Cytology of a mass on the meningeal surface of the left brain in a dog

An 11-year-old neutered male Labrador Retriever presented to Tufts University School of Veterinary Medicine for a 2-week history of seizures and altered behavior. Magnetic resonance imaging (MRI) revealed a homogeneously enhancing mass involving the surface of the left temporal, parietal, and occipital lobes of the brain, causing secondary hydrocephalus. Cytology samples obtained during rostrotentorial craniectomy contained abundant amorphous pink material suggestive of neuropil, scattered leukocytes, capillary fragments, large polyhedral nerve cell bodies, and other smaller cells with pale blue cytoplasm that occasionally were vacuolated and contained fine eosinophilic granules. The cytologic diagnosis was neoplasia, possibly meningioma. Ninety days after surgery the patient was euthanized after MRI results confirmed regrowth of the tumor. Histologic samples of the meningeal lesion collected at necropsy consisted of sheets and nests of loosely packed, large polygonal cells that compressed the brain parenchyma. The cytoplasm was eosinophilic and slightly granular, whereas nuclei were dense and eccentric. Neoplastic cells stained positive for S-100 protein, periodic acid-Schiff, and were partially diastase resistant. Vimentin staining was negative. Ubiquitin staining was light but diffusely positive. Ultrastructural features of the neoplastic cells included numerous secondary lysosomes and irregular pleomorphic nuclei. The final diagnosis was meningeal granular cell tumor. This case documents the cytologic and histologic features of an uncommon type of meningeal tumor.     

Sebaceous carcinoma of the salivary gland in a cat

Sebaceous carcinoma of the submandibular salivary gland is described in a cat, tumour cells were characterized histologically by moderate amounts of pale eosinophilic or vacuolated cytoplasm. Tumour cells were stained with antibody to cytokeratins (CKs 5. 6, 8, 17 and 19) and with lectins Con A and wheat germ agglutinin (WGA); this occurs in many other types of salivary gland tumour and is a feature of normal salivary gland acinar cells.     

Chordoma of the thoracic vertebrae in a Bengal tiger (Panthera tigris tigris)

A 19-year-old female Bengal tiger (Panthera tigris tigris) was presented with hind limb weakness, ataxia and respiratory distress. Computed tomography revealed a mass between the left side of the T7 vertebra and the base of the left 7th rib. The tiger then died, and necropsy was performed. Grossly, the vertebral mass was 6 × 5.7 × 3 cm, and invaded the adjacent vertebral bone and compressed the T7 spinal cord. Histologically, the mass was composed of large, clear, vacuolated and polygonal cells with osteochondral matrix. Cellular and nuclear atypia were moderate. The vacuolated cells stained positively for cytokeratin and vimentin and negatively for S-100. Based on these findings, the present case was diagnosed as a vertebral chordoma; the first report in a tiger.     

Invasive ductal carcinoma of the mammary gland in a mare

A 21-year-old thoroughbred mare had a 35 x 14 x 10 cm mass involving the mammary gland. Metastases were found in the kidneys, lungs, skeletal muscles, and regional lymph nodes. Histopathologic examination of the tumor revealed a ductal solid carcinoma with extensive intraductal and intralobular involvement and focal infiltration of the adjacent stroma. The intralobular neoplasms were divided into irregularly shaped islands and sheets of polygonal and spindle-shaped epithelial cells by thick or thin fibrous connective tissue bundles. The neoplastic cells had a small or moderate amount of cytoplasm that stained faintly with eosin and round or oval hyperchromatic nuclei. Immunohistochemically, the neoplastic cells were strongly positive for Lu-5, weakly positive for AE1/AE3, vimentin, and glial fibrillary acidic protein, and negative for cytokeratin 8, cytokeratin 14, alpha-smooth muscle actin, calponin, and S100. The neoplasm was diagnosed as an invasive ductal carcinoma of the mammary gland with multiple metastases.     

Morphology, cytochemical staining, and ultrastructural characteristics of the blood cells of the giant lizard of El Hierro (Gallotia simonyi)

The object of this study was to examine the erythrocytes, leukocytes and thrombocytes of the giant lizard of El Hierro (Gallotia simonyi) by light and electron (TEM) microscopy, and cytochemical staining. Smears were prepared from blood from the ventral coccygeal vein of 10 healthy adult lizards (five males and five females) from the Giant Lizard of El Hierro Reproduction and Research Centre, Canary Islands, Spain. The cytochemical stains used were: benzidine peroxidase (BP), chloroacetate esterase (CAE), alpha-naphthyl acetate esterase (ANAE), acid phosphatase (AP), periodic acid-Schiff (PAS), toluidine blue (TB) and May-Grünwald-Giemsa (MGG). Electron microscopy was also performed on all samples. Heterophils had granules that were heterogeneous in both size and electron density, and stained with BP, PAS and ANAE. Eosinophil granules were homogeneously electron-dense and stained for AP, CAE and ANAE. Basophils had both highly and moderately electron-dense granules, and stained with TB and ANAE. Azurophil granules were of low electron-density and stained for AP, CAE and ANAE. Azurophil cytoplasm was vacuolated on TEM. The cytoplasm of lymphocytes contained many ribosomes and was positive for AP. Monocytes had a large nucleus and a vacuolated cytoplasm but did not stain by any of the cytochemical methods used. Thrombocytes had a relatively large nucleus but little cytoplasm; they did not stain cytochemically. The blood cells of the giant lizards of El Hierro differ from those of other members of the Order Squamata both morphologically and cytochemically. The variation in cytochemical responses in the blood of reptiles makes it necessary to study species individually if meaningful clinical decisions are to be made.     

Histopathological and immunohistochemical characteristics of two canine lipid-rich mammary carcinomas

Clinicopathological and immunohistochemical findings of two uncommon canine lipid-rich mammary carcinomas are described. The predominant histological feature in both tumours was the presence of at least 80% of cells with intracytoplasmic vacuoles which stained positively with Sudan IV but not with alcian-blue periodic acid-schiff method. In both tumours, small groups of non-vacuolated cells were identified among the vacuolated cells. However, histological and immunohistochemical differences were also found between these tumours. Thus, one of them was composed of tumour cells with a large and single vacuole, which were arranged in lobular pattern, while the other neoplasm showed an intraductal growth of tumour cells with a fine vacuolated cytoplasm. Immunohistochemically, in the first tumour most vacuolated cells were positive for CK (cytokeratin)8-7, indicating a secretory epithelial immunophenotype while CK5 and CK8-7-expressing non-vacuolated cells were associated with luminal duct immunophenotype. However, in the second tumour the expression of CK14 in most of vacuolated cells and alpha-smooth muscle actin (alpha-SMA) in non-vacuolated cells, alone or in combination with CK5 suggested a myoepithelial immunophenotype for both cell types. These results suggest heterogeneity of the cell type and growth pattern for this type of canine tumour as has been described in women but not in dogs.     

Lesions of bone and bone marrow in myeloid leukosis occurring naturally in adult broiler breeders

Lesions of bone and bone marrow in myeloid leukosis (ML) occurring naturally in adult broiler breeders were investigated pathologically. During gross examination, nodules and protrusions were commonly observed on the surface of the sternum, ribs, vertebrae, and synsacrum. The bone marrow of all the bones of the body was pale in color. Histologically, granulated myelocytes proliferated in the bone marrow of various bones and in the periosteum of the sternum, ribs, vertebrae, and synsacrum. The first proliferation of tumor cells occurred in the bone marrow of epiphysis. The myelocytes invaded through haversian and Volkmann's canals from the bone marrow to periosteal areas. Hematopoiesis was suppressed by marked proliferation of tumor cells in the bone marrow of the whole bone. Atrophy was also seen in the bones, including medullary bones of the chickens suffering from ML. Proliferation of myelocytes was seen in the bone marrow and periosteum of ossified cartilaginous rings of the trachea and larynx. Marked proliferation of myelocytes was seen in the dura mater of spinal cords, and it subsequently depressed the spinal cords. Bone formation with cartilage was seen in the periosteum of the sternum having marked proliferation of myelocytes in the bone marrow and periosteum. Ultrastructurally, tumor cells showed large nuclei and cytoplasm with large round electron-dense lysosomes. The virus particles were rarely detected in the cytoplasm of tumor cells. The polymerase chain reaction test of tumor samples showed positive for subgroup J avian leukosis virus. This study indicates that the myelocytes can invade through the compact bones to the periosteum in the sternum, ribs, vertebrae, synsarcum, and ossified cartilage of trachea and larynx having thinner compact bones. In addition, the periosteal osteogenesis with cartilage in the sternum may be reactive change against the bone atrophy because of the marked proliferation of myelocytes.     

Cervical spinal chordoma with chondromatous component in a dog

A 7-year-old male Belgian Shepherd dog was presented with sudden onset of lateral recumbency and tetraparesis. At the level of the third cervical vertebra, magnetic resonance imaging demonstrated an intrameningeal and intramedullary mass lesion. The animal was subsequently euthanatized. A necropsy revealed a semitranslucent solid mass infiltrating dorsal and ventral dura mater and the spinal cord. Histologic examination revealed a lobulated pleomorphic mass, mainly resembling undifferentiated cartilage interspersed by spindle-shaped and polygonal cells with highly vacuolated cytoplasm (physaliphorous cells). Immunohistochemistry of the tumor cells demonstrated dual expression of vimentin and cytokeratin. Based on the histologic and immunohistochemical results, the diagnosis of a chordoma with chondromatous component was made.     

Immunohistochemical detection of the enzyme glutamic acid decarboxylase and hormones of the islets of Langerhans in spontaneous insulin-dependent diabetes mellitus in cattle

Immunohistochemical expression of glutamic acid decarboxylase (GAD) enzyme was detected in the pancreatic islets of 12 cattle with spontaneous insulin-dependent diabetes mellitus (IDDM). The most characteristic changes were atrophy and decreased number of pancreatic islets, enlarged islets with vacuolated beta cells, and lymphocytic islet adenitis. Atrophied islets were composed of small islet cells without cytoplasmic insulin-positive granules. Immunohistochemically, GAD was not found in the cytoplasm of atrophied islet cells. Furthermore, enlarged islets consisting of islet cells with vacuolated cytoplasm were frequently observed. The cytoplasm of vacuolated cells contained very few GAD- and insulin-positive granules, indicating beta cell destruction. Enlarged islets with mild lymphocytic infiltrates were frequently observed. These findings suggest that islet cells in cattle with IDDM lose their insulin synthesis function and their ability to regulate hormonal secretion of alpha and delta cells.     

Pathological, ultrastructural, and immunohistochemical changes caused by Lelystad virus in experimentally induced infections of mystery swine disease (synonym: porcine epidemic abortion and respiratory syndrome (PEARS))

The pathogenicity and pathogenesis of Lelystad virus was studied in six 6-day-old SPF piglets. A third passage of the agent was propagated on porcine alveolar macrophages and intranasally inoculated into pigs. Pigs were killed at hours 24, 48, 60, and 72, and on days 6 and 8 after inoculation. From day 2 on pigs developed diffuse interstitial pneumonia with focal areas of catarrhal pneumonia, and from this day on splenic red pulp macrophages were enlarged and vacuolated. Lelystad virus was re-isolated from the lungs of infected pigs from day 2 after inoculation. Lelystad virus antigens were detected by immunohistochemical techniques in bronchiolar epithelium and alveolar cells, and in spleen cells of infected pigs from day 2 after inoculation. Ultrastructural examination of tissues by electron microscopy revealed degenerating alveolar macrophages and epithelial cells in lungs and nasal mucosa, with excessive vacuolation of the endoplasmic reticulum. Although the respiratory tract seems to be the target organ for this virus, macrophages in other organs, such as the spleen, can also be infected. This preference for macrophages may impair immunological defences.     

Feline cystic thymoma: a clinicopathologic,immunohistologic, and electron microscopic study of 14 cases

Cystic thymoma was diagnosed in 14 cats in a period of 6 years. The most common clinical sign was laboured breathing. The tumours were characterized by various-sized cystic spaces with central vessels. The epithelial cells varied from oval to spindle to polygonal cells enclosing cystic spaces or in pure epithelial cell components. The nuclei of the neoplastic cells had scattered chromatin and small nucleoli. The cytoplasm was pale eosinophilic. The concentration of mature lymphocytes varied from area to area with rare germinal centres. Immunohistochemically, the epithelial cells stained only with AE(1)/AE(3). The central vessels were positive for vimentin, smooth muscle actin, and factor VIII antigen. Electron microscopy revealed that the cyst walls were lined by epithelial cells that were joined by desmosomes, and the walls were well separated from the cystic spaces by a well-defined basement membrane. The neoplastic epithelial cells contained scattered tonofilaments. Three of the cats had metastasis to the lymph nodes and lungs. Two novel cases of ectopic cystic thymoma have also been described. Results of this study reveal that cystic thymoma is uncommon in cats, and that the histomorphologic, immunohistochemical, and electron microscopic features are similar to those of cystic thymoma in humans.     

Time of appearance of esterase-positive alveolar macrophages in bovine fetal lung

Bovine fetal lung tissue was examined histochemically, using alpha-naphthyl butyrate (nonspecific method), to detect the presence of esterase-positive pulmonary alveolar macrophages (PAM). Positively stained PAM were first seen in the alveolar septa and lumina of the lungs of 4 of 8 fetuses 240 days old. Macrophages (n = 100) in 240-day-old fetuses were 6.74 +/- 0.07 microm, and in most cells, esterase-positive granules were sparsely distributed in the cytoplasm. In 10 fetuses 250 days old, the alveolar septa and lumina of lungs contained positively stained macrophages, as did those of all older fetal lungs examined. Compared with macrophages of the 240-day-old fetuses, those (n = 50) of 250-day-old fetuses were significantly larger, 7.32 +/- 0.10 micron (P less than 0.0001), and esterase-positive granules were heavily distributed throughout the cytoplasm. In all fetal lungs, T lymphocytes contained a single, small, esterase-positive granule, compared with the numerous diffuse esterase-positive granules in PAM. Type-II pneumonocytes with well-developed lamellar bodies had no evidence of positive nonspecific esterase reaction in lungs.     

Chronic granulocytic leukaemia/ eosinophilic leukaemia in a dog?

A suspected case of chronic granulocytic leukaemia (chronic myelogenous leukaemia) with eosinophil differentiation and hepatic involvement is described in a 3–5-year-old rottweiler dog. Neoplastic eosinophilic infiltrates were also present in the lungs, kidneys, mesenteric lymph nodes and peribronchial and prester-nal lymph nodes. Both pleural and abdominal effusions contained predominantly eosinophils and their precursors. The bone marrow showed an increase in the myeloid to erythroid ratio and an increase in the marrow granulocyte reserve. The majority of cells identified within the bone marrow were of the eosinophil series. Myelocytes and promyelocytes predominated.