Bovine leukemia virus-associated B cell lymphoma with severe pleomorphism in a steer.

We examined a 26-month-old steer with neoplastic lesions in the spleen, lymph nodes, heart and kidneys, characterized by pleomorphic lymphoid cells that were immunohistochemically positive for CD20. The presence of bovine leukemia virus (BLV) at >200,000 copies per 100,000 cells by quantitative RT-PCR was considered to be due to random integration of the provirus into the neoplastic cells´ genomes. Inverse PCR identified the presence of one, two, two and three different malignant clones in the heart, spleen, mesenteric node and blood, respectively. Because BLV can rapidly induce lymphoma and a high proviral load facilitates B-cell carcinogenesis, multiclonal tumor development was suspected in the present case.

A porcine lymphoma-derived cell line co-expressing IgM, IgG and IgA.

A cell line (PL38PB) was established from blood samples of a 6-month-old pig that was diagnosed with lymphoma with CD5 expression. Histopathological examination revealed neoplastic lesions in the spleen, liver and lymph nodes. Tumor cells were immunohistochemically positive for CD20 and immunoglobulin heavy chains (µ, γ and α). Membranous CD5 and cytoplasmic Immunoglobulin M (IgM), ​Immunoglobulin G (IgG) and ​Immunoglobulin A (IgA) were detected in PL38PB cells by flow cytometry. In addition, the cytoplasm of PL38PB cells were positive for IgM, IgG and IgA by immunofluorescent. However, no Ig secretion was detected in culture supernatant by Ouchterlony gel diffusion method. Results suggest that PL38PB cells express three Ig isotypes that are produced but not secreted.

Histological comparison of malignant epithelioid mesothelioma in young and adult cattle.

The histology and immunohistochemistry of pleomorphic and conventional epithelioid mesotheliomas were examined. The former was detected in two young calves aged 2 and 4 months and was characterized by pleomorphic and atypical cells with decreased expression of cytokeratin 7 (CK7). In contrast, the latter was found in a 31-month-old heifer, consisting of tumor cells uniform in size and shape with CK7 expression in nearly all cells. Production of collagen by tumor cells was demonstrated in both histological types, and was considered to be characteristic of bovine epithelioid mesothelioma. Pleomorphic mesothelioma is far more pleomorphic and mitotically active than conventional mesothelioma, and its normal counterpart may be immature mesothelial cells with high proliferation potential, which exist in fetal life and early calfhood.

Combined hepatocellular-cholangiocarcinoma in a cow.

We examined a 10-year-old cow in which about half of the liver was displaced by malignant tissue consisting of hepatocellular carcinoma (HCC) and cholangiocarcinoma (CC). Cytokeratin (CK) 18 and 7 were expressed in the latter. Metastasis was present in the hepatic, pancreaticoduodenal and mediastinal lymph nodes, where malignant cells had hepatocellular features, but more pleomorphic and atypical than in the primary lesion. Areas composed solely of CC cells or less-differentiated HCC cells were observed. In contrast, well-differentiated HCC cells were almost always admixed with the other two types, and may have had the ability to transform into CC cells and to dedifferentiate into less-differentiated cells. This report suggests that CK18 is an excellent marker for biliary differentiation in cattle.

Laryngeal squamous cell carcinoma and papilloma associated with Equus caballus papillomavirus 2 in a horse.

A case of laryngeal squamous cell carcinoma (SCC) and squamous papilloma in a 19-year-old Thoroughbred stallion is described. The animal exhibited severe wheezing caused by laryngopharyngeal stenosis. Histological examination identified laryngeal, laryngotracheal, and guttural pouch tumor masses consisting of areas of SCC. In the epiglottic lesion, the overlying epithelium was replaced by papilloma cells, and superficial cells frequently had nuclear inclusion bodies that expressed oncoprotein E6, which is characteristic of high risk human papillomaviruses. The papillomatous epithelium was continuous with epithelium composed of SCC cells. Equus caballus papillomavirus 2 (EcPV2) DNA was detected in the guttural pouch tumor. These findings suggest that laryngeal SCC and papilloma are a continuum of EcPV2-induced neoplastic lesions in horses.

Erythroblastic sarcoma in the thoracic cavity of a cow.

Erythroblastic sarcoma in a 10-year-old Japanese Black cow with anemia is described. Tumor masses or nodules were located mainly in the thoracic cavity, and some lymph nodes were slightly enlarged. Although neoplastic involvement of the bone marrow was detected, the cow was not leukemic. The diagnosis was made based on the localized distribution of neoplastic lesions, no increase of intravascular nucleated cell number, deeply eosinophilic cytoplasm in some tumor cells, and frequent immunoreactivity of the tumor cells for hemoglobin. The tumor cells were characterized by marked pleomorphism and atypia; such morphological deviation from their normal counterparts may be connected with functional deviation resulting in the sarcomatous growth of these erythroid cells.

Detection of bovine papillomavirus type 2 DNA in calf conjunctival myofibroblastoma.

An 8-month-old male Japanese Black calf was referred for the evaluation of a slow-growing conjunctival mass in the right eye. A superficial keratectomy was performed followed by recurrence on two occasions. No metastases were found in surrounding tissues. Histological, immunohistochemical, and ultrastructural investigation revealed that both the primary and the recurrent lesions were benign, conjunctival, myofibroblastomas. Interestingly, bovine papillomavirus type 2 (BPV-2) DNA was detected in both myofibroblastoma lesions. Archival bovine myofibroblastomas from the vulva and neck were also analyzed for papillomaviral genomes. BPV-2 DNA was also amplified from these lesions. To the best of our knowledge, this is the first report describing a potential causal relationship between BPV-2 infection and conjunctival myofibroblastoma.

Survival times with and without tube feeding in patients with dementia or psychiatric diseases in Japan.

BACKGROUND: It is widely supposed that there has been no evidence of increased survival in patients with advanced dementia receiving enteral tube feeding. However, more than a few studies have reported no harmful outcome from tube feeding in dementia patients compared to in patients without dementia. METHODS: This was a retrospective study. Nine psychiatric hospitals in Okayama Prefecture participated in this survey. All inpatients fulfilling the entry criteria were evaluated. All subjects suffered from difficulty with oral intake. Attending physicians thought that the patients could not live without long-term artificial nutrition. The physicians decided whether to make use of long-term artificial nutrition between January 2012 and December 2014. RESULTS: We evaluated 185 patients. Their mean age was 76.6 ± 11.4 years. Of all subjects, patients with probable Alzheimer's disease (n = 78) formed the biggest group, schizophrenia patients (n = 44) the second, and those with vascular dementia (n = 30) the third. The median survival times were 711 days for patients with tube feeding and 61 days for patients without tube feeding. In a comparison different types of tube feeding, median survival times were 611 days for patients with a nasogastric tube and more than 1000 days for those with a percutaneous endoscopic gastrostomy tube. CONCLUSION: Patients with tube feeding survived longer than those without tube feeding, even among dementia patients. This study suggests that enteral nutrition for patients with dementia prolongs survival. Additionally, percutaneous endoscopic gastrostomy tube feeding may be safer than nasogastric tube feeding among patients in psychiatric hospitals.

Long-Term Survival of Patients Receiving Artificial Nutrition in Japanese Psychiatric Hospitals.

BACKGROUND/AIMS: Most patients with dementia suffer from dysphagia in the terminal stage of the disease. In Japan, most elderly patients with dysphagia receive either tube feeding or total parenteral nutrition. METHODS: In this study, we investigated the factors determining longer survival with artificial nutrition. Various clinical characteristics of 168 inpatients receiving artificial nutrition without oral intake in psychiatric hospitals in Okayama Prefecture, Japan, were evaluated. RESULTS: Multiple logistic regression analysis showed that the duration of artificial nutrition was associated with a percutaneous endoscopic gastrostomy (PEG) tube, diagnosis of mental disorder, low MMSE score, and absence of decubitus. CONCLUSION: Patients with mental disorders survived longer than those with dementia diseases on artificial nutrition. A PEG tube and good nutrition seem to be important for long-term survival.

Eosinophilic granuloma with Splendore-Hoeppli material caused by Mannheimia granulomatis in a calf.

A large subcutaneous mass, formed on the left lower jaw of a 4-month-old Japanese Black male calf, was partially excised for histological and bacteriological examinations. Antibiotic treatment resulted in a good prognosis. Bacteria isolated from the excised material were characterized by weak hemolysis and positive reactions for catalase and oxidase, and were 99% identical to Mannheimia granulomatis strains. The presence of the leukotoxin gene product was demonstrated by polymerase chain reaction amplification. Histological examination showed that the excised material was composed of dense fibrous connective tissue with sparsely distributed eosinophilic granulomas or abscesses. These foci frequently contained Splendore-Hoeppli material with rod-shaped Gram-negative bacteria. Except for the absence of lymphangitis and the presence of basophils and mast cells, the histology of this lesion resembled that of lechiguana associated with coinfection of M. granulomatis and Dermatobia hominis. Leukotoxin was demonstrated by immunohistochemistry within Splendore-Hoeppli material and was judged to be responsible for its formation.

Immature T cell neoplasms in three young cattle.

Immature T cell neoplasms in three young Holstein cattle with neoplastic involvement of the thymus are described. Case 1, with a precursor T lymphoblastic leukemia (calf form of leukosis), was an 86-day-old female calf. The leukemia was characterized by replacement of the bone marrow and spleen by leukemia cells, but preservation of epithelial frameworks throughout the thymus. The other two neoplasms were thymic γδ T cell lymphomas, which were observed in a 246-day-old steer (case 2) and a 16-month-old heifer (case 3). Histological examination revealed obliteration of the normal thymic architecture and stromal fibrosis, with the spleen and liver far less severely affected than in case 1. There were cytological differences bewteen the tumors in case 1 and cases 2 and 3. Additionally, WC1 and CD8 were expressed only in the latter. Thus, the leukemia and these lymphomas should be regarded as independent disease entities on the basis of histological and immunohistochemical characteristics.

Intranuclear coccidiosis caused by Cyclospora spp. in calves.

Intranuclear coccidiosis in 3 calves is described. Two calves with no genetic relationship were kept in the same pen, and the third calf was on another farm. The animals suffered from watery diarrhea or severe emaciation. Histologic examination showed epithelial desquamation (cases 1 and 2) or atrophy (case 3) of the jejunal villi. Coccidial meronts or merozoites were present in the nuclei of the majority of villus epithelial cells. There were rare intranuclear macrogametocytes, macrogametes, microgametocytes, microgametes, and oocysts in cases 1 and 2, but these were more easily observed in case 3. Parasite 18S ribosomal RNA sequences from case 1 showed 99.5% sequence identity with Cyclospora sp. Guangzhou 1, which has been found in fecal samples of cattle from China. In addition to the molecular results in one of the cases, the fact that the microgametocytes and oocysts were noticeably smaller in size than those of Eimeria alabamensis in all cases indicates the close association between Cyclospora and the 3 cases described herein.

Eosinophilic granuloma with Splendore-Hoeppli material caused by toxigenic Corynebacterium ulcerans in a heifer.

Raised lesions were present on the left nasal vestibule of a 20-month-old Japanese Brown heifer. The largest mass which caused partial nasal obstruction was removed surgically. Corynebacterium ulcerans was identified in the mass. 16S ribosomal RNA and RNA polymerase beta subunit genes were 100% and 98% identical to other C. ulcerans strains. Histologically, multiple foci of eosinophilic granuloma with Splendore-Hoeppli material were seen. Rod-shaped Gram-positive organisms were detected with metachromatic granules, producing diphtheria toxin with 5, 30 and 48 amino acid differences to another C. ulcerans strain, C. diphtheriae or C. pseudotuberculosis, respectively. The toxin is highly cytotoxic and may be responsible for the formation of abundant Splendore-Hoeppli material. The lesion was therefore judged to be an allergic reaction to bacterial antigens or diphtheria toxin.

Signet ring cell lymphoma with plasmacytic differentiation in a pig.

A case of signet ring cell lymphoma in a 3-year-old mixed-breed sow is described. Macroscopical examination revealed enlargement of superficial, thoracic and abdominal lymph nodes and multiple tumor masses in the liver. The neoplastic tissue was composed of follicle center-like structures, in which neoplastic cells with Russell bodies were conspicuous. The bodies were immunostained for IgM (κ), and corresponded to moderately dense amorphous material within markedly distended cisternae of rough endoplasmic reticulum (RER) at the ultrastructural level. In contrast to typical signet ring cell lymphoma, the component cells of which resemble follicular center B lymphocytes with poorly developed RER, most neoplastic cells had features of plasma cells characterized by a cartwheel arrangement of heterochromatin and development of RER. Signet ring cells frequently had one or a few large Russell bodies occupying the entire cytoplasm, which may have been caused by abundant synthesis and defective secretion of immunoglobulin.

Pneumocystis carinii Infection in a domestic goat (Capra hircus domesticus) with multibacillary paratuberculosis.

Pneumocystis carinii (P. carinii) is an opportunistic fungal pathogen commonly found in many mammalian host species, but rarely in goats. A 3-year-old, female, Tokara-native-goat (Capra hircus domesticus) died of apparent malnutrition caused by multibacillary paratuberculosis. While inflammatory response was slightly observed in the respiratory organs, P. carinii trophozoites and cysts were immunohistochemically observed in the pulmonary alveoli of the infected animal. P. carinii specific DNA was amplified from the formalin fixed and paraffin embedded lung samples. Molecular phylogenetic analyses of the mitochondrial large subunit ribosomal RNA region of P. carinii revealed genetic divergence from previously described P. carinii isolates from other mammalian host species. This is the first description of concurrent infection with P. carinii and the Mycobacterium avium subspecies paratuberculosis in a domestic goat.

Mast cell sarcoma with megakaryocytic differentiation in a calf.

A case of mast cell sarcoma in a 5-month-old Holstein female calf is described. Macroscopically, enlargement of the spleen, lymph nodes, tonsils and kidneys was noted, and there were tumor masses in the neck region and on the pleura and peritoneum. The pericardium and uterine and ureter walls were also involved by tumor. Most neoplastic cells had eosinophilic granules, which were metachromatic and positive for naphthol AS-D chloroacetate esterase and tryptase, whereas smaller numbers of cells were positive for factor VIII-related antigen, a marker of megakaryocytes. Some of the predominant type of these tumor cells were found within the epithelia of the lungs, tonsils, gastrointestinal tract, liver, ureters, urinary bladder and uterus. Their normal counterparts were considered to be globule leukocytes.

Lymphoid neoplasms in swine.

Seventeen cases of lymphoid neoplasms in swine were investigated and divided into eight histological types. Cases 1-3 were precursor B lymphoblastic leukemias, which occurred in three piglets from the same dam. Cases 4 and 5 were diagnosed, respectively, as a precursor B lymphoblastic lymphoma and a thymic B cell lymphoma, because there were cytological differences between the lymphomas. These five cases of immature B cell malignancies expressed CD79a and terminal deoxynucleotidyl transferase (TdT). Mature B cell lymphomas were divisible into follicular (case 6), diffuse centroblastic (case 7) and intestinal large B cell (cases 8-11) lymphomas. Unlike in case 7, the neoplastic cells in cases 8-11 showed cytological features intermediate between centroblasts and immunoblasts. The mature lymphomas were characterized by positive immunolabeling for CD79a and cytoplasmic immunoglobulins. A case of thymic γδ T cell lymphoma (case 12) were positive for CD3, CD5, WC1 and TdT. Instead of TdT, perforin was expressed in γδ T cell lymphomas (cases 13-17), whose histological characteristics were epitheliotropism, homing into T cell zones of lymphatic tissues, and cytological atypia and pleomorphism. In the present study, lymphoid neoplasms could be classified into discrete histological types, some of which were considered to be specific for swine.

Characterization of novel bovine papillomavirus type 12 (BPV-12) causing epithelial papilloma.

Bovine papillomavirus type 12 (BPV-12, putative type BAA1) was detected in epithelial papilloma located on the tongue of an infected cow. Then, the whole genome was sequenced, and phylogenetic analysis illustrated that it should be classified as a member of the genus Xipapillomavirus. The viral genome is 7197 base pairs in length and contains five early ORFs (E1, E2, E4, E7 and E8), three late ORFs (L1, L2 and L3), and a long control region that possesses replication regulatory elements. Meanwhile, mRNA of each gene was detected in the papilloma sample. The papilloma was identified as epithelial papilloma by histological and immunohistochemical examination. Based on the genome information and pathological properties, BAA1 was designated as BPV-12 in this study.

Acute myeloid leukemia with mastocytic and megakaryocytic differentiation in a calf.

A diagnosis of acute myeloid leukemia was made in a 10-month-old Holstein female calf. The leukemia was macroscopically characterized by great enlargement of the spleen and moderate enlargement of some lymph nodes. Histochemical and immunohistochemical examination disclosed the presence of neoplastic cells either containing metachromatic and tryptase-positive granules or expressing factor VIII-related antigen. The granules, which were positive for naphthol AS-D chloroacetate esterase and did not have particulate contents, were distinct from those of basophilic leukemia cells. This leukemia was thought to be derived from a common myeloid progenitor capable of giving rise to megakaryocyte-erythrocyte progenitors and granulocyte-monocyte progenitors with the ability to differentiate into mast cells.

Cytologic variants of γδ T cell lymphoma in cattle.

Two cytologic variants of γδ T cell lymphoma are described. Case 1 represented a giant cell variant found in a 5-year-old Holstein cow, which had large tumor masses in the pelvic cavity. This variant consisted of very large lymphoid cells with round to oval nuclei, medium-sized nucleoli and abundant cytoplasm. Case 2 was an aborted 7-month-old female Holstein fetus, which represented an immature cell variant. Most of the neoplastic lesions were located in the skin and pleural and peritoneal submesothelial tissues. The neoplastic tissues were composed of homogeneous growth of lymphoma cells characterized by inconspicuous nucleoli and finely dispersed chromatin. Both cases demonstrated CD3, CD8 and WC1 immunoreactivity. The current study revealed that there are 4 cytologic variants (common, giant cell, hypergranular and immature cell) in bovine γδ T cell lymphomas.