BACKGROUND: Hypocalcaemia as one of the most important ovine metabolic diseases occurs during late gestation and early lactation. OBJECTIVES: The aim of this study was to investigate the physiological effects of experimentally induced hypocalcemia on serum biochemical, enzymatic and electrolyte changes. Methods: Ethylene diamine tetra-acetic acid (EDTA) solution (4.6 %) was intravenously infused to 5 healthy sheep (experimental group). Meanwhile, 5 healthy sheep received 0.9% saline solution (IV) and kept at similar condition as control group. In both groups serum was collected before and after EDTA infusion at different time points. Aspartate transaminase (AST), alanine transminase (ALT), glucose, blood urea nitrogen (BUN), total protein, calcium (Ca) and inorganic phosphorus (Pi) levels were measured. RESULTS: Clinical signs of hypocalcemia and/or changes in heart rate was caused by EDTA infusion. Serum calcium levels showed significant decrease (39.2% at its maximum state) following EDTA infusion (p<0.05), compared to the pre-infusion state. Pi levels showed significant decrease (57.8% at its maximum state) during 1 to 4 hours following EDTA infusion. Meanwhile, it was accompaning with an increase (9.9% at its maximum state) at 5.5 to 22 hours after EDTA infusion. Increase in serum glucose levels during 1 to 3 hours following EDTA infusion was determined as much as 60% (38.44 mg/dl) at its maximum state. An increase (51.1% at its maximum state) was shown in blood urea nitrogen levels after EDTA infusion in experimental group. Changes in total protein, ALT and AST were not significant before and after EDTA infusion. CONCLUSIONS: While, experimental hypocalcemia can be accompanied with a decrease in serum Ca level, time dependent alterations in Pi and an increase in glucose and BUN levels can be observed. However, AST, ALT and total protein values can be without any changes.