Abstract. The hematopoietic cells in blood and/or bone marrow from 20 leukemic dogs and 22 control dogs were characterized using a battery of cytochemical stains. The results of cytochemical staining led to modification of the diagnoses based on clinical, hematologic and histologic findings in seven (35%) of the leukemias. Sudan black B and chloroacetate esterase served as granulocytic markers in both the control and leukemic groups. Peroxidase activity was present in the granulocytes and monocytes of control animals but not the blasts of leukemic dogs. Alkaline phosphatase-positive staining of granulocytic precursors was a consistent finding in granulocytic and myelomonocytic leukemia, and alkaline phosphatasepositive lymphoblasts were seen in 38% of lymphocytic leukemias. Diffuse alpha naphthyl butyrate esterase-positive staining marked monocytes in both control and leukemic dogs. Cytochemical staining was found to be a valuable diagnostic aid in the classification of leukemias in the dog. Materials and MethodsBone marrow specimens were aspirated from the proximal humerus or femur from 20 leukemic dogs presented to the OSU Veterinary Teaching Hospital. Samples also were aspirated from 22 control dogs, including dogs with non-hematologic diseases. In some cases, blood samples collected on the same day as the marrow aspirate also were stained. A few samples were obtained from a private veterinary laboratory. Blood films and bone marrow coverslip smears were prepared and air-dried-generally within one hour of sample collection. Films were stained with Wright-Giemsa, alpha naphthyl butyrate estera~e,'~ and periodic acid-Schiff." Cytochemical staining kits (Sigma Chemical Co., St. Louis, Mo.) were used 36 for Sudan black B, acid phosphatase with and without tartrate, alkaline phosphatase, and chloroacetate esterase procedures. Positive staining was defined as distinct staining of a minimum of 10% of the abnormal cells. A positive control was included in each batch of stains. If staining could not be done within eight hours, the films were stored unfixed at room temperature in the dark with the exception of the slides for acid phosphatase and alkaline phosphatase which were fixed in accordance with the manufacturer's instructions (Sigma Chemical Co., St. Louis, Mo.) and stored at 4°C. The corresponding histologic sections of representative tissues from dogs that were necropsied along with the Wright-Giemsa blood and bone marrow films were reviewed by one of the authors (GK). Diagnosis was based on cell morphology, anatomic distribution of lesions, and histologic findings. To simplify data analyses, the lymphocytic leukemias were subdivided into lymphoblastic leukemia," lymphoblastic leukemia associated with lymphoma,22 and well-differentiated lymphocytic leukemia' on morphologic criteria only. These results were compared with the corresponding cytochemical findings.
Abstract. Hematopoietic cells in blood and/or bone marrow from 23 leukemic cats and ten control cats were characterized using a battery of cytochemical enzyme stains. The results of cytochemical staining led to modification of diagnosis based on clinical, hematologic and histopathologic findings in four (1 7%) of the leukemias. Sudan black-B and chloroacetate esterase served as granulocytic markers in both the control and leukemic groups. Peroxidase activity was seen in the granulocytes and monocytes of control animals but not in the blasts of leukemic cats. Diffuse alpha naphthyl butyrate esterase staining marked monocytes in both control and leukemic animals. Cytochemical staining was found to be a valuable aid in the classification of leukemias in the cat. Materials and MethodsBone marrow specimens were aspirated from the proximal humerus or femur from leukemic cats presented to the Ohio University Veterinary Teaching Hospital. Samples were also collected from ten control cats including patients with nonhematologic diseases. In some cases, blood samples collected on the same day as the marrow aspirate also were stained. A few samples were obtained from a private veterinary laboratory. Blood films and bone marrow coverslip smears were prepared and air-dried generally within one hour of sample collection. Films were stained with Wright-Giemsa, alpha naphthyl butyrate esterase," and periodic a~id-Schiff.~~ Cytochemical staining kits (Sigma Chemical Company, St. Louis, MO) were used for the Sudan black-B, acid phosphatase with and without tartrate, alkaline phosphatase, and chloroacetate esterase procedures. The chloroacetate esterase procedure was modified by increasing the incubation temperature to 37 C.A Dositive control was included in each batch of stains. Posphatase and alkaline phosphatase which were fixed in accordance with the manufacturer's instructions (Sigma Chemical Company, St. Louis, MO) and stored at 4 C. The corresponding histologic sections of representative tissues from animals that were necropsied along with the blood and bone marrow films were reviewed by one of the authors. Diagnosis was based on cell morphology, anatomic distribution of lesions, and histologic findings. '3,'5,L8,19,28 Due to the paucity of specific diagnostic criteria for feline leukemias, the lymphocytic leukemias were subdivided only into lymphoblastic and well-differentiated lymphocytic categories.28 These results were compared with the corresponding cytochemical findings. ResultsThe cytochemical staining characteristics of normal cells (Table 1)
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