Haemangioma (HA) and haemangiosarcoma (HSA) are among the most common splenic neoplasms in dogs. The survival time in splenic HSA is short, probably due to the lack of proper biological markers allowing early detection. We investigated the serum angiopoietin-2 (Ang-2) concentrations in 9 healthy dogs and 40 dogs with abnormal splenic masses. The Ang-2 concentration differences were further compared in healthy dogs, dogs with splenitis, splenic HA and HSA. The results showed that the Ang-2 level in healthy dogs was significantly lower than in the splenitis and splenic HA cases. Moreover, the Ang-2 level was significantly higher in splenic HA than in splenic HSA. Conversely, no significant differences in Ang-2 level were recorded between healthy and splenic HSA dogs, and between splenitis and splenic neoplasms (HA and HSA). No significant correlations were observed between the Ang-2 level and (i) the clinical stage, (ii) histological growth pattern, and (iii) median survival time of splenic HSA dogs. In conclusion, serum Ang-2 concentration is a potentially useful biological marker for the discrimination of dogs with splenitis and splenic HA, as well as for the differentiation of splenic HA from its malignant form, HSA.