Disseminated peripheral neuroblastoma in a Rhodesian Ridgeback dog.

CASE REPORT: A 4-year-old neutered male Rhodesian Ridgeback dog with right-sided Horner's syndrome, bilateral laryngeal paralysis, neck pain and bilateral hindlimb ataxia was euthanased following deterioration of its neurological status. Necropsy examination revealed an off-white retropharyngeal neoplastic mass (100 × 30 × 30 mm) attached to the base of the skull on the right side and macroscopic nodular metastases in the spleen and three vertebral bodies (C6, C7 and T6), including a nodule attached to the dura at C7. Histological evidence of neuroblastic tumour was detected in these macroscopic lesions, a regional lymph node, bone marrow of a femur and all 15 vertebral bodies (C1-T8) examined, including the three with macroscopic metastases, and in the lumens of small blood vessels in the lungs and liver. Ganglion cell differentiation was detected only in the primary retropharyngeal mass, one splenic nodule and the C7 dural nodule. Neoplastic cells were immunoreactive to neurofilament protein (ganglion cells only), vimentin and synaptophysin, and were negative for S100 protein, GFAP, CD3 and Pax5. CONCLUSION: The diagnosis was disseminated peripheral neuroblastoma, differentiating subtype (International Neuroblastoma Pathology Classification), with likely primary involvement of the right cranial cervical ganglion. This appears to be the first report of neuroblastoma in a dog with widespread occult haematogenous metastasis to bone marrow.

Peripheral blood abnormalities and bone marrow infiltration in canine large B-cell lymphoma: is there a link?

Official guidelines do not consider bone marrow (BM) assessment mandatory in staging canine lymphoma unless blood cytopenias are present. The aim of this study was to find out if blood abnormalities can predict marrow involvement in canine large B-cell lymphoma. BM infiltration was assessed via flow cytometry. No difference was found between dogs without haematological abnormalities and dogs with at least one. However, the degree of infiltration was significantly higher in dogs with thrombocytopenia, leucocytosis or lymphocytosis and was negatively correlated to platelet count and positively to blood infiltration. Our results suggest that blood abnormalities are not always predictive of marrow involvement, even if thrombocytopenia, leucocytosis or lymphocytosis could suggest a higher infiltration. BM evaluation should therefore be included in routine staging in order not to miss infiltrated samples and to improve classification. However, its clinical relevance and prognostic value are still not defined and further studies are needed.

Malignant mesenchymoma with widespread metastasis including bone marrow involvement in a dog.

A male castrated Golden Retriever was presented for evaluation of a large mass over the left shoulder extending to the lower part of the neck that had been present for an extended period of time, but had a recent history of rapid growth. Previous aspirates of the mass were consistent with a lipoma. The mass was surgically excised and was diagnosed as an extraskeletal osteosarcoma based on histopathology. After surgery, the dog was initiated on a chemotherapy protocol with carboplatin and metronomic cyclophosphamide. He became neutropenic, anemic, and thrombocytopenic 14 days after the carboplatin treatment was administered. The neutropenia resolved, but the anemia and thrombocytopenia progressed. A bone marrow aspirate revealed erythroid hypoplasia, myeloid hyperplasia with a predominance of early precursors, and a subset of cells that made up 20% of the total population that were reported as bizarre and unclassifiable. These cells were discrete in nature and were thought to be hematopoietic in origin. The dog was euthanized due to deterioration of the clinical condition. On postmortem examination, widespread metastasis involving the lungs, liver, kidney, heart, and bone marrow was found. Histopathology of the tumor lesions determined 2 distinct malignant populations of liposarcoma and osteosarcoma, consistent with malignant mesenchymoma. However, the possibility of 2 separate neoplastic processes cannot be definitively excluded. This is the first report of bone marrow metastasis of a malignant mesenchymoma in a dog.

Detection of carcinoma micrometastases in bone marrow of dogs and cats using conventional and cell block cytology.

BACKGROUND: The presence of human breast carcinoma micrometastases in bone marrow is associated with poor overall survival, poor breast-cancer-specific survival, poor disease-free survival, and poor distant disease-free survival. In veterinary practice, the detection of micrometastases as a component of clinical staging is a routine practice for lymphomas and mast cell tumors, but not for carcinomas. OBJECTIVES: This prospective study evaluated whether the identification of micrometastases from various carcinomas in dogs and cats in bone marrow using cell block cytology is technically feasible and whether it correlates with routine cytologic examination. METHODS: Thirteen dogs and 4 cats with various types of carcinomas were available for analysis. Routine and cell block cytologic evaluation combined with immunocytochemical staining with antibodies to CKAE1/AE3 and CK7 were performed on all bone marrow samples. RESULTS: Bone marrow micrometastasis was demonstrated by both methods in 2 dogs with advanced disease. In one case cells were immunoreactive for both CKAE1/AE3 and CK7. CONCLUSIONS: This study demonstrates that cell block cytology is a practical and useful method for bone marrow evaluation and is suitable for cytokeratin immunocytochemical analysis.

Mammary gland carcinoma in a dog with peripheral blood and bone marrow involvement associated with disseminated intravascular coagulation.

A 7-year-old female Leonberger dog was referred to the National Veterinary School of Lyon Teaching Hospital with a 2-day history of anorexia and bleeding. A mammary mass had been removed 7 months earlier, but histologic examination was not performed. On physical examination, the dog was depressed and had pale mucous membranes and numerous petechiae and hematomas. Significant laboratory findings were moderate thrombocytopenia, prolonged prothrombin, activated partial thromboplastin, and thrombin times, hypofibrinogenemia, and increased concentration of fibrin(ogen) degradation products. A peripheral blood smear, buffy coat preparation, and bone marrow aspirate contained low numbers of large atypical cells that had moderate nuclear:cytoplasmic ratios, oval nuclei with multiple prominent nuclei, and basophilic cytoplasm with villous projections. A small nodule was found in the left inguinal mammary gland, and a fine-needle aspirate contained cells similar to those in blood and bone marrow. In samples of blood, bone marrow, and the mammary mass, the neoplastic cells were immunoreactive for cytokeratin. The diagnosis was mammary carcinoma with secondary disseminated intravascular coagulation (DIC) and disseminated tumor cells in bone marrow and circulating tumor cells in blood; this diagnosis was not confirmed by histopathologic examination. Owing to clinical deterioration and the poor prognosis, the dog was euthanized and a necropsy was not performed. This is the first report of a canine mammary carcinoma with circulating tumor cells and secondary DIC.

Eosinophilic granulocytic sarcoma in a pig.

A case of granulocytic sarcoma originating from an eosinophilic lineage is described in a 5-year-old, mixed-breed, female pig. The pig had been originally sent to slaughter in a good plane of nutrition and without displaying clinical disease. At gross examination, green masses were observed in several bones, especially vertebrae, sternum, pelvis, and long bones such as femur and humerus. Similar masses were seen in skeletal muscles, lymph nodes, and kidneys. Cytology revealed large numbers of round cells with round nuclei and scant cytoplasm (myelocytes); some of these cells had a fine eosinophilic granularity to their cytoplasm (eosinophil myelocytes). Histologically, the neoplastic cells formed sheets that completely obliterated the normal architecture of subperiosteal bone marrow. The cytoplasm of the neoplastic cells stained strongly by Sirius red stain of eosinophil and was positively marked by immunohistochemistry using an anti-myeloperoxidase antibody. The association of gross examination, cytology, histology, histochemistry, and immunohistochemistry findings is consistent with a diagnosis of eosinophilic granulocytic sarcoma.

Primary nasal histiocytic sarcoma of macrophage-myeloid cell type in a cat.

A 16-year-old neutered male Burmese cat was presented with a locally invasive nasal mass. The cytological and histological findings on incisional biopsy of this mass were suggestive of histiocytic sarcoma. Tumour cells expressed CD18, major histocompatibility complex class II, lysozyme and alpha-naphthyl acetate esterase; and lacked expression of CD3, CD79a, CD1a, CD1b, calprotectin, CD11c and E-cadherin. These findings are consistent with a myeloid-macrophage lineage. Metastasis to the bone marrow was present on necropsy examination. Histiocytic sarcoma should be considered in cats presented with primary round cell neoplasia of the nasal cavity.

Metastatic melanoma appearing to originate from the beak of a racing pigeon (Columba livia).

A 12-year-old male pigeon (Columba livia) was reported to have a small lesion on the lower beak, which was diagnosed as malignant melanoma. After surgery to remove it, the general condition of the bird worsened and the bird died. Post mortem examination revealed gross, black-colored lesions in the liver, kidney, spleen, and femur bone marrow. Lesions were not found in the adrenals. Histopathologically, the neoplastic cells had a large amount of melanin pigment and showed severe anisokaryosis and prominent nucleoli. Mitotic activity was infrequent but varied considerably where it was observed.

Acute megakaryoblastic leukemia in a German Shepherd dog.

An 11-year-old spayed-female German Shepherd dog was presented to the Veterinary Medical Teaching Hospital at Kansas State University with a history of weight loss, anorexia, depression, and lethargy for 2-3 weeks. Radiographic examination revealed a mass in the spleen and several round radiodense foci in the liver. CBC results included normocytic normochromic anemia, marked thrombocytopenia, and low numbers of neoplastic cells that frequently had cytoplasmic projections or blebs. A bone marrow aspirate contained about 80% neoplastic megakaryoblasts with the same microscopic features as those observed in peripheral blood. Using flow cytometry, cells of large size were identified in peripheral blood that expressed CD41/61, CD45, CD61, and CD62P (P-selectin) and were negative for markers of T cells, B cells, monocyte/macrophages, and dendritic cells. Because of the poor prognosis, euthanasia and subsequently necropsy were performed. On histopathologic examination, neoplastic megakaryoblasts were identified in spleen, liver, mesenteric lymph node, and the pulmonary vasculature. Using immunohistochemistry, the neoplastic megakaryoblasts weakly expressed von Willebrand factor. Based on microscopic and immunophenotypic findings, a diagnosis of acute megakaryoblastic leukemia (AMegL) was made. To our knowledge, this is the first report of AMegL in a domestic animal in which immunophenotyping by flow cytometry and a panel of antibodies against CD41/61, CD61, and CD62P were used to support the diagnosis.

Disseminated melanoma in a dog with involvement of leptomeninges and bone marrow.

An 11-year-old, Black and Tan Coonhound dog was presented with a history of lameness of the right hind leg for 2 months, osteolysis in the right distal femur, a pulmonary mass, and a presumptive diagnosis of osteosarcoma. By cytologic examination, neoplastic melanocytes were noted from fine needle aspirates of the femoral and pulmonary masses. Postmortem examination revealed a disseminated melanoma involving the right femoral bone marrow, lung, multiple lymph nodes, and adrenal gland, with diffuse infiltration of the leptomeninges of the brain and spinal cord. This case report describes a unique presentation of canine melanoma, which in some ways resembles leptomeningeal melanomatosis, a rare human melanoma variant.

Clinicopathological features and outcome for dogs with mast cell tumors and bone marrow involvement.

BACKGROUND: Mast cell tumors (MCTs) with bone marrow (BM) involvement are poorly documented in dogs and are associated with a poor prognosis. Successful treatment strategies have not been described. HYPOTHESIS: Clinicopathologic findings of affected dogs are not specific. Administration of lomustine or imatinib is beneficial. ANIMALS: Fourteen dogs with MCT and BM involvement. METHODS: Clinical and laboratory evaluations were performed in each dog on admission and during follow-up. All dogs received prednisone. Additionally, 8 dogs received lomustine and 3 dogs received imatinib. Imatinib was administered if tumor-associated tyrosine kinase KIT was aberrant. RESULTS: On admission, 11 dogs had a single cutaneous nodule and 3 dogs had multiple nodules. Involvement of regional lymph nodes, liver, or spleen was observed in each dog. BM infiltration with mast cells (MCs) was observed in all dogs. On CBC, nonregenerative anemia, leukopenia, or thrombocytopenia was common. Four dogs had circulating MCs. Increased alkaline phosphatase or alanine transferase activity was observed in 12 and 10 dogs, respectively. Treatment with lomustine induced partial remission in 1 of 8 dogs. Median survival time was 43 days (range, 14-57). Dogs on imatinib experienced complete remission. Two dogs survived for 117 and 159 days, and the third was alive after 75 days. Dogs treated symptomatically did not improve and were euthanized after 1, 14, and 32 days. CONCLUSIONS AND CLINICAL IMPORTANCE: A combination of clinical and laboratory evaluation helps in identifying dogs with MCT and BM infiltration. Administration of lomustine is not helpful in affected dogs. The beneficial effect of imatinib warrants further investigation.

Cytosine arabinoside in addition to VCAA-based protocols for the treatment of canine lymphoma with bone marrow involvement: does it make the difference?

Cytosine arabinoside (ara-C) is a component of many protocols for the treatment of acute leukaemia and non-Hodgkin lymphomas in humans. The aim of the study was to prospectively evaluate the efficacy of ara-C in a myeloablative regimen in a cohort of canine lymphomas with bone marrow involvement. Seventeen dogs were enrolled. Eight were treated with a VCAA-based protocol (Group 1) and nine with the same regimen added with ara-C (Group 2). Ara-C was administered on a 5-day schedule as an i.v. continuous infusion at the dose of 150 mg m(-2) per day for five consecutive days. During treatment complete remission (CR) was achieved in two dogs in Group 1 and in eight dogs in Group 2. CR rate was significantly higher in Group 2 (P < 0.01). Median survival was 72.5 days (range 6-174) in Group 1 and 243 days (range 73-635) in Group 2. Survival was significantly longer in Group 2 (P < 0.001). Both protocols were well tolerated, with a low incidence of adverse events. Ara-C added to a VCAA-based protocol appears to be safe and beneficial in dogs with stage V lymphoma. Incorporation of the nucleoside analogue might be crucial for the development of future therapeutic strategies in dogs.

A peripheral primitive neuroectodermal tumor with generalized bone metastases in a puppy.

A peripheral primitive neuroectodermal tumor (pPNET), most consistent with a human Ewing's sarcoma, is described in a 5-month-old male Australian Shepherd puppy. The first tumor site detected was in the left frontal bone of the skull with apparent subsequent rapid metastases to multiple sites in the axial and appendicular skeleton and bone marrow, kidneys, and perihyphophyseal meninges. Radiographically, all bone lesions were lytic and there was also a humeral bone fracture. Histologically, the tumor was diagnosed as a small round blue cell tumor. At this stage, the differential diagnosis included a lymphoma, rhabdomyosarcoma, and a PNET of the peripheral nervous system. However, the cells had positive expression of triple neurofilament antigens as detected immunocytochemically. The cells were negative for a broad panel of canine-specific leucocyte cell marker antigens for desmin, smooth muscle actin, synaptophysin, and CD99. Ultrastructurally, the cells contained occasional dense core neurosecretory granules and intermediate filaments with intercellular desmosomal-like junctions and abundant glycogen clusters. Based on the age of the dog, the clinical history, the distribution of gross lesions, histologic characteristics of a small round blue cell tumor, and immunocytochemical and ultrastructural evidence of neuroectodermal differentiation, a diagnosis of a pPNET similar to a human Ewing's sarcoma was made.

Immunocytological confirmation of bone marrow metastases in a dog with cholangiocarcinoma.

The case history of a four-year-old, male Bernese mountain dog is presented. Carcinoma cells were detected in the liver by ultrasound-guided fine-needle aspiration. Bone marrow aspirated from the iliac crest and the left femur showed a distinct infiltration by carcinoma cells. Immunocytological examination of the liver and bone marrow metastases showed a negative staining result for large spectrum cytokeratin (CK) KL1, a strong positive result for CK7 and a focal weak positive result for CK20. The dog was euthanased due to the grave prognosis. Histopathological examination revealed metastatic cholangiocarcinoma. The authors conclude that cytological and immunocytological examination of bone marrow aspirates should be used more frequently for the detection of distant metastases of carcinomas in small animal medicine.

Extradural spinal, bone marrow, and renal nephroblastoma.

A 1-year-old, female intact Shetland sheepdog presented with acute onset of neurological signs. Physical examination revealed a large abdominal mass. Neurological examination revealed multifocal disease with neck pain, short-strided forelimbs, and hind-limb paresis with loss of tail and anal tone. Blood work, imaging techniques, cytopathology, and histopathology led to a diagnosis of renal, bone-marrow, and extradural spinal nephroblastoma. This report documents potential clinical and pathological manifestations of canine nephroblastoma that have not been previously reported.

Cytology of bone marrow.

Cytologic examination of bone marrow aspirates can provide a wealth of diagnostic information. Practitioners should not hesitate to perform bone marrow aspirates when indicated. This article is designed to assist the practitioner in the evaluation of bone marrow aspiration biopsies. The indications for marrow evaluation, methods of sample collection, sample preparation, and cytologic examination of bone marrow are discussed. Cases are provided to demonstrate accurate interpretation of bone marrow aspirates.