Blood samples were obtained from the dorsal coccygeal vein and the brachial vein of five adult (four females and one male) and two subadult males of marginated tortoise (Testudo marginata) and hematologic and biochemical parameters were compared. Significant differences were found for red blood cell count, hematocrit, hemoglobin concentration, total proteins, uric acid, aspartate aminotransferase, alanine aminotransferase, lactate dehydrogenase, alkaline phosphatase, calcium, and phosphorus, which were greater in the brachial vein samples. Hemodilution due to lymph was observed when collecting blood from the dorsal coccygeal vein, and it is thought to be the cause of the differences found. This research documented that the brachial vein is a more reliable and consistent venipuncture site than dorsal coccygeal vein in marginated tortoise.