Comparison of molecular and growth properties for two different canine distemper virus clusters, Asia 1 and 2, in Japan.

To compare the molecular and growth properties of two newly isolated canine distemper virus strains in the Asia 1 and 2 groups with clinico-pathological findings in dogs, nucleotide and predicted amino acid sequence comparisons of genes H and P were performed together with comparative growth profiling. The predicted amino acid sequences of the H gene contained 12 cysteine residues that were conserved among the examined Asia 1 and Asia 2 viruses. The hydrophobic region in the H gene of the Asia 2 isolates was one amino acid longer than that of the Asia 1 group. The H gene of the Asia 1 group had nine putative asparagine (N)-linked glycosylation sites, while there were eight sites in the Asia 2 group. The titers of the cell-associated viruses for the Asia 1 strains were higher than those of the released viruses and were opposite to those of the Asia 2 strains in a previous study. The molecular and growth properties of the Asia 1 and Asia 2 groups seem to vary, although no significant differences were observed in the clinical signs and pathological findings between the two groups.

Immunohistochemical studies on cytokeratin 8 and 18 expressions in canine cutaneous adnexus and their tumors.

The expressions of cytokeratin 8 and 18 (CK8 and CK18) in the normal canine skin (2 cases) and cutaneous adnexal tumors (127 cases) were investigated immunohistochemically. In the normal skin, co-expression of CK8/18 was found in the glandular epithelium of apocrine sweat glands, and single CK8-immunoreactivity was detected occasionally in the external root sheath at the isthmus and suprabulbar regions of the hair follicles. Neoplastic glandular epithelial cells in all apocrine gland tumors (21/21 cases, 100%) had co-expression of CK8/18. In trichoblastomas (27/28 cases, 96%), most neoplastic cells were diffusely positive for CK8, but those were negative for CK18. Single CK8-expression was also observed in basaloid neoplastic cells in several cases of trichoepitheliomas (7/19 cases, 37%) and pilomatricoma (1/7 cases, 14%). In several cases of trichoblastomas (4/28 cases, 14%) and trichoepitheliomas (2/19 cases, 11%), tumor cells forming glandular structures had co-expression of CK8/18. There were no positive reactions for both CK8 and 18 in infundibular keratinizing acanthomas, and sebaceous and hepatoid gland tumors. The present findings indicate that co-expression of CK8/18 is a specific feature of apocrine sweat glands and single CK8-expression represents the natures of external root sheath or pluripotential stem cells. Thus, the combination of CK8- and 18-immunostainings may have the utility to confirm the directions of differentiation in canine cutaneous adnexal tumors providing a reliable hallmark for histopathological diagnoses.

Establishment and characterization of a cell line, MCO-Y4, derived from canine mammary gland osteosarcoma.

A cell line, MCO-Y4, was established from a mammary gland osteosarcoma of a 16-year-old female mongrel dog. Histopathologically the tumor was composed of osteoblastic cells with an osteoid meshwork and chondroid matrix. The mean doubling time of the cells at the 93rd passage was 32.39+/-4.66 hr. Immunohistochemically, the osteoblastic and chondroblastic cells were positive for bone morphogenetic protein (BMP)-2/4 and BMP receptor (BMPR) II. The cultured cells were spindle in shape during the growth and the confluent phases. No tumor matrix was detected in the culture dish by alcian blue staining or von-Kossa silver impregnation. MCO-Y4 cells on the chamber slides showed intense immunoreactivity for BMP-2/4 and BMPR II. Noggin, an antagonist for BMP-2/4, showed the growth inhibition on MCO-Y4 cells. In addition, fibronectin might be potential for stimulating growth of MCO-Y4 cells. When transplanted into severe combined immunodeficiency mice, the cells formed tumors consisting of solid proliferation of osteoblastic and fibroblastic cells with woven-bone trabeculae. These tumor cells were intensely positive for BMP-2/4 and BMPR II. Our results suggested that the cell line might be useful for studying the role of BMPs in canine osteosarcoma and the mechanism of ossification.

A case of glomus tumor in a dog.

A subcutaneous mass at the digit of the left-hind limb of a 12-year-old, male mongrel dog was examined. A white firm mass, approximately 1 x 2 cm in diameter, was excided surgically. Histopathologically, the mass formed multiple nodules consisting of mixed proliferation of round epithelioid cells arranged in cord or sheet-like structures and small spindle cells forming loose irregular bundles. The epithelioid cells often showed proliferation around the blood vessels. A few giant cells scattering in the neoplastic foci were observed. The neoplastic cells were positive for alpha-smooth muscle actin and vimentin, and were negative for cytokeratin (AE1/AE3), desmin, factor-VIII related antigen, S-100 protein, and neuron specific enolase. On the basis of these findings, this tumor was diagnosed as glomus tumor. Since the present neoplasm had neither recurrence nor distal metastasis during the 12 month after surgical resection, the biological natures of the present neoplasm are supposed to be benign.

Co-localization of chondromodulin-I (ChM-I) and bone morphogenetic protein-6 (BMP-6) in myoepithelial cells of canine mammary tumors.

To compare the roles of chondromodulin-I (ChM-I) and bone morphogenetic protein-6 (BMP-6) in ectopic mesenchymal tissue formation in canine mammary gland tumors, 33 tumors and 2 normal mammary glands were examined. Immunohistochemical analysis revealed co-expression of ChM-I and BMP-6 in canine mammary tumors. In mixed tumors, newly formed woven bone with ossified cartilage matrix was observed in 4/9 cases. The osteoblasts lining the woven bone showed moderate immunoreactivity to ChM-I and BMP-6. Almost all chondrocytes and proliferative myoepithelial cells within the basement membrane showed intense immunoreactivity to both, and the myoepithelial cells adjacent to the mature cartilage showed the most intense immunoreactivity. The immunoreactivity to ChM-I and BMP-6 of the interstitial myoepithelial cells in the myxomatous stroma varied in each focus of mixed tumors. Similar findings were found in complex adenomas. In simple adenomas, hyperplasic myoepithelial cells within the basement membrane showed moderate immunoreactivity to both markers. Western blot analysis detected a 25 kDa band of ChM-I in fresh tissue samples from three mixed tumors. Our results support the hypothesis that proliferating myoepithelial cells with ChM-I and BMP-6 expression play important roles in mesenchymal metaplasia in canine mammary tumors.

Investigation of initial changes in the mouse olfactory epithelium following a single intravenous injection of vincristine sulphate.

To investigate initial changes in the olfactory epithelium, vincristine sulphate (VCR) was administered intravenously once to male BALB/c mice on day 1 in comparison with unilateral bulbectomy (UBT). The light and electron microscopy of the olfactory epithelium, nerve and/or bulb with BrdU-morphometry was performed sequentially. Further, whole-body radioluminography was conducted at 1 and 24 hours postdose. Apoptosis and an increased number of mitotic cells with a tendency toward decreasing BrdU-positive olfactory epithelial cell counts were observed in olfactory epithelial cells at 6 hours postdose of VCR and became more pronounced at 24 hours postdose. These changes disappeared on days 4 or 15, but minimal axonal degeneration was seen in the olfactory nerve from day 4 onward. Semiquantitative measurement of VCR levels in the ethmoturbinals elicited high drug retention even 24 hours after administration. In contrast, UBT showed no effect on mitosis and BrdU-positive cell counts at 6 hours postdose, but severe lesions in the olfactory epithelium and nerve were seen on days 2, 4, and/or 15. The above results suggest that the initial event of VCR-induced apoptosis in the mouse olfactory epithelium would be mitotic arrest with high drug retention, unlike that evoked by UBT.

A neurological disease with spongy degeneration in a newborn Japanese black calf.

A Japanese Black calf, 3 day-old male, showed severe ataxia, lateral recumbency, and opisthotonos at the birth. Histopathological examinations revealed severe status spongiosis throughout the central nervous system. Numerous vacuoles within the neuropile varying in size and shape were observed in both formalin-fixed paraffin and cryostat sections. In the lesions, a limited number of spheroids and macrophages were observed within the myelin sheaths with very mild astrogliosis. These vacuoles were negative for both periodic acid Schiff and Sudan black stains. The clinical and histopathological features were almost in conformity with those of bovine maple syrup urine disease (MSUD). Although we could not confirm completely the etiology, congenital hereditary neurological diseases including MSUD are considerable as the possible disease entry in the present case.

Ability of the Hershberger assay protocol to detect thyroid function modulators.

In vivo screening methods for detection of thyroid function modulators are now under development in many research laboratories. We assessed the applicability of the Hershberger assay protocol to screen for thyroid function modulators. In experiment 1, castrated male BrlHan WIST@Jcl (GALAS) rats were administered a potent thyroid peroxidase inhibitor, 3-amino-1,2,4-triazole (AT), in doses of 0, 40, 200, and 1,000 mg/kg/day with gravimetric endpoint, and in experiment 2, castrated and intact male rats were administered in doses of 0, 40, and 200 mg/kg/day, with quantification of the extent of hypertrophy of the thyroid epithelium, to assess the effects of castration, by gavage to 8-week-old for 10 consecutive days. At necropsy of both experiments, the thyroid glands and hypophysis were collected and fixed with 10% neutral-buffered formalin. To avoid crushing during weighing because of their fragility, the thyroid glands and hypophysis were weighed approximately 24 h after fixation with 10% neutral-buffered formalin. All animals were sacrificed approximately 24 h after the final dose. In experiment 2, the thyroid glands of all animals were stained with hematoxylin and eosin for histological examination and morphometry of follicular epithelial height. In experiment 1, absolute and relative thyroid weights in all of the AT groups were statistically increased in a dose-dependent manner, regardless of the testosterone propionate (TP)-injection. In experiment 2, the results showed a significant increase in thyroid weight in the 200 mg/kg groups of both castrated and intact rats. Hypophyseal weight was unaltered by AT, but comparison of vehicle-treated groups showed that the hypophyseal weight of the castrated rats was greater than that of the intact rats. Enlarged thyroid glands were observed in the AT-treated rats at necropsy. Histological examination of the thyroid glands of all the AT-treated animals showed hypertrophy and hyperplasia of the follicular epithelial cells, and the height of follicular epithelium of the thyroid glands increased in a dose-dependent manner in both the castrated and intact rats. In experiment 1, assessment of the (anti-) androgenic action of AT in seminal vesicle weight revealed a significant increase in the 200 and 1,000 mg/kg + TP groups in a dose-dependent manner. These results suggest that the effect of AT can be detected by the Hershberger assay 10-day administration protocol and may be useful for screening for thyroid function modulators regardless of whether the animals have been castrated.

The growth profiles of three types of canine distemper virus on Vero cells expressing canine signaling lymphocyte activation molecule.

To know growth profiles of canine distemper virus (CDV) on Vero cells stably expressing canine signaling lymphocyte activation molecule (Vero-DogSLAMtag; Vero-DST cells), the propagation of three strains of CDV was tested in Vero-DST cells in comparison with parental Vero cells. Strain MD77 could grow well in both cell lines, but demonstrated no syncytium formation or indistinguishable rounding cytopathic effects (CPE) in Vero cells. Strains Onderstepoort and KDK-1 also grew well in Vero-DST cells with apparent syncytium CPE, while they grew less or no efficiently, respectively, in Vero cells. All three CDV strains demonstrated the peak titers, in Vero-DST cells before reaching to an extensive CPE and drastic decrease of titers at/after full CPE. Immunohistochemistry revealed that viral antigens of all CDV strains were found exclusively in the syncytia in Vero-DST cells, while in Vero cells, viral antigen was identified in their single cells for strain MD77 but none for other strains. Thus, every strain of CDV could grow well in Vero-DST cells and behaved differently against Vero cells. These results would be of practical value for workers of CDV because 1) In Vero-DST cells, by observation of distinct syncytium CPE, the highest titer or the best growth of virus could be identified; 2) In Vero cells, various CDV strains could be readily classified after propagation in Vero-DST cells.

Preliminary evaluation of an in utero-lactation assay using 6-n-propyl-2-thiouracil.

In this preliminary study, the potential of an in utero-lactation assay to detect thyroid effectors was evaluated by treating three dams/group with 6-n-propyl-2-thiouracil (PTU), a known thyroid antagonist, by oral gavage at doses of 0, 0.0032, 0.016, 0.08 and 0.4 mg/kg/day during fetal organogenesis and lactation. Hearing disturbances and an elevated relative thyroid weight were observed in offspring of both sexes in the 0.4 mg/kg/day group. The Biel-type water T-maze test showed an increase in the number of errors made by females in the 0.4 mg/kg/day group. Histopathologically, flattening of follicular epithelium, a decrease in resorptive colloid droplets, degeneration of follicular epithelium, and hyperplasia of follicular epithelium were observed in males belonging to the 0.4 mg/kg/day group. Histopathological abnormalities were also observed in some offspring belonging to the 0.08 mg/kg/day group. In the dams, hypertrophy of the follicular epithelium of the thyroid was observed in the 0.4 mg/kg/day group. Although we could not explain the mechanism for the difference in the effects seen in the offspring and the dams, the effect of PTU in utero through lactational exposure is apparently different from that resulting from exposure in homeostatically mature rats. Most reports studying PTU have involved administration in water or in food, and reports on the oral gavage of PTU during the fetal organogenesis and lactation period are very rare. This assumes that dosages >0.4 mg/kg/day would also produce clear anti-thyroid effects by oral gavage and, possibly, emphasizes that dosages <0.4 mg/kg/day did not have a noticeable effect. Based on the present results, a study to determine the reproducibility of the data in a much larger number of dams will be performed to confirm the findings in the present study, and to evaluate other endpoints, such as hormonal evaluation of dams and their offspring, sexual developmental landmarks, and fertility of the offspring.

Growth characteristics of canine distemper virus in a new cell line CCT cells originated from canine malignant histiocytosis.

Canine distemper virus (CDV) growth and the morphological characterization were examined in a cell line established from a canine malignant histiocytosis (CCT cell line). The susceptibility of the CCT cells to 3 CDV strains, FXNO, YSA-TC and MD-77 was shown by detection of the antigen in the indirect fluorescent assay. After passaging 4 and 9 times through the CCT cells, only FXNO strain could produce the syncytia where demonstrated the antigens. Titers of 9 passaged viruses through the CCT cells showed slightly higher in the CCT cells than those in Vero cells. Morphological characterization of karyorrhexis and specific DNA ladder by extracted DNA electrophoresis indicated apoptosis in the CDV infected CCT cells.

Olfactory epithelial lesions induced by various cancer chemotherapeutic agents in mice.

In order to examine and compare the potential toxicity in the olfactory epithelium, the antitumor drug vincristine sulfate (VCR), vinblastine sulfate(VBL), vindesine sulfate (VDS), paclitaxel (PTX), mitomycin C (MMC), 5-fluorouracil, (5-FU) or cisplatin (CDDP) was intravenously injected once(designated as day 1) at an estimated 10% lethal dose (LD(10)) to male BALB/c mice. The animals were necropsied on days 2, 5 and 15, and nasal tissues were examined by light-microscopy, counting of epithelial cells positive for terminal deoxyribonucleotidyl transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL), immunohistochemical staining with keratin antibody, and electron microscopy. Further, to delineate the drug disposition in the target organ, whole-body radioluminography was performed 1 hour and 24 hours after treatment with the LD(10) of PTX or 5-FU. Of the antitumor drugs employed, only the antimicrotubule agents, VCR, VBL, VDS, and PTX, induced single cell death in the olfactory epithelium, especially sensory cells on day 2, atrophy of the olfactory epithelium on day 5, and myelin fragmentation in the trigeminal nerve on day 15. PTX induced the strongest changes among the 4 antimicrotubule agents. The cell death was confirmed to be apoptosis by TUNEL assay and electron microscopy, whereas the change in horizontal basal cells of the olfactory epithelium was shown not to be apoptosis by keratin staining. In quantitative radioluminography,radioactivity of PTX in the nasal tissues both 1 hour and 24 hours after administration was about 4- or 5-fold higher than those of 5-FU. These results suggest that tubulin-targeting antitumour drugs could induce apoptosis in the olfactory epithelial cells of mice and that high drug distribution may effect the onset of the olfactory lesions.

Lymphoid apoptosis in acute canine distemper.

The relationship between the canine distemper virus (CDV) infection and apoptosis in the canine lymphoid tissues was investigated using immunostaining for single stranded DNA (ssDNA), TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method, and electron microscopy. Twenty-six lymphoid tissues from 8 spontaneously CDV-infected dogs and 1 non-infected dog were used, and lesions were classified into 4 groups according to frequency of the CDV-antigen. Histologically, the degree of lymphoid depletion tended to depend on amount of CDV antigen. The numbers of ssDNA- and TUNEL-labeling cells were significantly high in the lymphoid tissues with abundant viral antigen. However, ssDNA- and TUNEL-positive lymphocytes were also frequently found even in the lymphoid tissues where there was only a small amount of CDV-antigen in sinus histiocytes. The incidence and distribution of apoptotic cells in the CDV-antigens-negative lymphoid tissues from infected dogs were equal to those from a non-infected dog. Double labeling immunostaining using a ssDNA and a CDV nucleocapsid protein (CDV-NP) antibody revealed that there were ssDNA positive but CDV-NP negative cells besides those stained doubly positive. Ultrastructurally, lymphocytes in the CDV-infected lymphoid tissues frequently had characteristic morphological features of apoptosis such as apoptotic bodies. All these results suggest that CDV leads to lymphocytic apoptosis directly or indirectly, resulting in severe lymphoid depletion and immunosuppression in acute or subacute phase of CDV infection.

A comparative pathological study on canine necrotizing meningoencephalitis and granulomatous meningoencephalomyelitis.

Canine necrotizing meningoencephalitis (NME) and granulomatous meningoencephalomyelitis (GME) were compared pathologically. Gross observation exhibited lateral ventricular dilation and discoloration, malacia and/or cavitation of the cerebrum in NME. On the contrary, gross changes were milder in GME, except for occasional visible granulomatous mass formation. Histopathologically, the lesions of NME were distributed predominantly in the cerebral cortex and various degrees of inflammatory and necrotic changes were observed according to clinical stages. Besides, microscopic lesions of GME were mainly distributed in the white matter of the cerebrum, cerebellum and brainstem, which are characterized by perivascular cuffing, multiple granulomas and leptomeningeal infiltrates. Although macrophages and lymphocytes were predominant in the inflammatory lesions of both disorders, macrophages in GME transformed into epithelioid cells and exhibited more massive infiltration. Although lectin RCA-1-reactive cells were numerous in both disorders, lysozyme immunoreactive cells in NME were fewer than that in GME. Parenchymal infiltration of MAC387-positive cells was common in GME and limited in NME. The number of CD3-positive lymphocytes in the GME lesions tended to be greater than in NME, though the difference was not statistically significant. Morphological and immunohistochemical differences of the lesions, in particular, the characteristics of infiltrative macrophages may reflect these different pathogeneses of the two disorders.

Three cases of canine gastrointestinal stromal tumors with multiple differentiations and c-kit-expression.

Three canine gastrointestinal stromal tumors (GISTs) were examined. Histopathologically, the tumor mass in the jejunum (Case 1) consisted of the proliferation of epithelioid cells with abundant eosinophilic or vacuolated cytoplasm. Gangliocyte-like or multinucleated giant cells were scattered. The tumor cells exhibited neural natures mimicking human gastrointestinal autonomic nerve tumors, which were immunopositive for several neuronal markers. Another jejunal mass (Case 2) was composed by a solid proliferation of spindle-shaped cells, arranging in interlacing fascicles and occasional storiform pattern. The tumor seemed to be classified undifferentiated GISTs, that showed no apparent neural or muscular features by ultrastructural and immunohistochemical examinations. In the pyloric mass (Case 3), the spindle cells having eosinophilic processes and elongated nuclei were arranged in sheets. Immunohistochemically, the tumor cells showed muscular natures as regards alpha smooth muscle actin and desmin expression.

Age-related histological changes in the canine substantia nigra.

Age-related changes in the canine substantia nigra, were examined using immunostaining for tyrosine hydroxylase (TH), glial fibrillary acidic protein (GFAP), neurofilament (NF), ubiquitin, single stranded DNA (ssDNA), and alpha-synuclein (alphaSN). Brain sections from 34 necropsied dogs, ranging from 2 months to 18 years old, were used for this study. On general histological examinations, several age-related changes, including lipofuscin deposition, polyglucosan bodies, amorphous basophilic inclusions and eosinophilic crystal inclusions, were found in the aged dogs. Immunohistochemically, TH-positive neurons were located only in the substantia nigra. The number of TH-positive neurons was well preserved in all dogs examined, however, the ratio of TH-positive neurons to GFAP-positive glial cells tended to show slight decrease in aged dogs. By ssDNA immunostaining for apoptotic cells, there were no significant results in the number of ssDNA-positive neurons. The number of ubiquitin- and NF-positive swollen neurites was increased markedly in aged dogs. Ubiquitin immunostaining revealed a small number of basophilic and eosinophilic inclusions, although both types of inclusions were negative for NF. By alphaSN immunostaining, no neurons were immunoreactive and no basophilic or eosinophilic intracytoplasmic inclusions were revealed. These results indicate that in the substantia nigra of aged dogs the dopaminergic neurons are well preserved, but intracytoplasmic inclusions and ubiquitin-positive degenerative neurites are commonly found.

Peripheral neuroblastoma in a young labrador retriever.

A 2-year-old Labrador Retriever developed atrophy of the right temporal muscle, subsequently showed generalized seizure and died 2 months after the clinical onset. Postmortem examination revealed the tumor masses in the right mandibulopharyngeal area, nasopharynx and intracranial space. Histopathologically, these tumor masses were composed of small round neoplastic cells and neuropil-like stroma separated by fibrovascular septa. In the neoplastic masses, small neoplastic cells with round to oval hyperchromatic nuclei and scanty cytoplasm predominated, and angulated neoplastic cells with larger nuclei and moderate cytoplasm were scattered. Immunohistochemically, neoplastic cells were positive for neuron specific enorase, neurofilament protein, chromogranin A, synaptophysin and tyrosine hydroxylase. Based on these findings, this case was diagnosed as peripheral neuroblastoma, presumably originated from the sympathetic ganglion, maybe right craninal cervical ganglion.

Ganglioglioma in the thalamus of a puppy.

A solitary brain mass of a 4-month-old miniature dachshund showing seizure-like neurological signs was examined histopathologically. At necropsy a white tumor mass, replacing the thalamus, approximately 1.5 cm in diameter, was found. There was cystic space filled with yellowish pale fluid in the central area of the tumor mass. Histopathological examination revealed that the mass consisted of irregularly arranged well-differentiated neuronal and glial cells, and multifocal mineral deposits. The neuronal cells had a large clear nucleus and various amount of Nissl substances in the cytoplasm. Some neural cells were bi-nucleated. Neither mitotic figures nor proliferating cell nuclear antigen (PCNA)-positive nuclei was found in the neuronal cells. Immunostaining for glial fibrillary acidic protein (GFAP) revealed diffuse proliferation of GFAP-positive glial cells and their processes, while these glial cells did not show apparent cellular atypism, mitotic activity, or PCNA-immunoreactivity. Accordingly, the present tumor was diagnosed as ganglioglioma, and hamartomatous histogenesis might be possible.

A comparative pathological study on granulomatous meningoencephalomyelitis and central malignant histiocytosis in dogs.

Histiocytic proliferative disorders in canine central nervous system (CNS) including granulomatous meningoencephalomyelitis (GME) and malignant histiocytosis were compared pathologically. Lesions of GME mainly existed in the white matter of the cerebrum, brainstem and cerebellum and consisted of characteristic perivascular cuffing, parenchymal granuloma and leptomeningeal infiltrates of mononuclear cells. In malignant histiocytosis, there were two histological patterns, diffuse proliferation of neoplastic histiocytes through the leptomeninges and neoplastic nodule formation in the parenchyma. Neoplastic histiocytes exhibited mild to severe cellular atypia and high ability of invasion into the brain parenchyma. Mitotic and phagocytic figures were also observed. Several histiocytic markers, including lysozyme, alpha1-antitrypsin and lectin RCA-1, revealed histiocytic origin of both inflammatory and neoplastic cells, however, those were not determinative for the discrimination between GME and malignant histiocytosis. CD3- and PCNA-positive cells existed in the lesions of both diseases. The number of CD3-positive cells in GME tended to be greater than in malignant histiocytosis, while the difference was not statistically significant.

Thymoma and multiple thymic cysts in a dog with acquired myasthenia gravis.

An anterior mediastinal cystic lesion in an 11-year-old mongrel dog was examined. The dog showed dysbasia and vomiting due to megaoesophagus, and anterior mediastinal round mass lesion, approximately 35 mm in diameter, was found by X ray. Based on clinical examinations, the dog was diagnosed as acquired myasthenia gravis and was successfully controlled by anticholinesterase treatment for approximately 4 months. The dog died of thermic stroke and was necropsied. Grossly, fatty tissues with cysts containing yellowish fluid and white nodules were found in the anterior mediastinal area. Histopathologically, multiple cysts, neoplastic tissues, and atrophic thymus were found within the examined tissues. The cysts were lined by thin wall consisting of ciliated long cuboidal and non-ciliated round cells and were filled with eosinophilic colloidal fluid. Some extended cysts contained neoplastic foci within their lumen and walls. The neoplastic tissues consisted of mixed population of large epithelial cells with abundant clear cytoplasm and large oval nuclei, and lymphocytes. Immunohistochemically, proliferating epithelial cells were intensely positive for keratin and cytokeratin, and more than half number of infiltrating lymphocytes were intensely positive for CD3 suggesting T cells. All these findings indicate the neoplastic lesion is thymoma and multiple cysts are considered as thymic or brachial cleft cysts.