SUMMARYA macrocytic hypochromic anemia with marked reticulocytosis was observed at each of 10 examinations during a 4 month period of a male Beagle dog that was 11 months old when first examined. The owner had noticed pale mucous membranes from the time the dog was purchased at 7 weeks of age. Based on the clinical history and negative LE, ANA and Coombs tests, an intrinsic erythrocyte defect was suspected.Erythrocyte osmotic fragility and hemoglobin electrophoresis were normal. Autohemolysis of incubated erythrocytes was marked. Hemolysis was uncorrected by glucose but partially corrected by ATP. With the exception of pyruvate kinase (PK), all of the erythrocyte enzymes measured had greater activities than those of control Beagle erythrocytes. In comparison to moderately reticulocyte‐rich controls. PK activity was low in the affected Beagle. Erythrocyte phosphoenolpyruvate content was increased significantly as expected in PK deficiency. The kinetic constant (Km) for phosphoenolpyruvate was lower than that of normal control dogs, a finding in agreement with studies of PK from deficient Basenjis. On these bases a diagnosis of PK deficiency was mad