This study aimed to determine the levels of homocysteine and cardiac markers in cows with hypocalcemia. This research was conducted with a total of 20 cows, including 10 with hypocalcemia and 10 healthy controls. Hypocalcemia was diagnosed by determining low blood calcium (Ca) levels, and a standard hypocalcemia treatment was initiated in those cows with the disease. Blood samples were taken before the treatment and again 12 h after the treatment, and there was no significant difference in the homocysteine levels between the groups. Ca, ionized calcium (ICa), magnesium (Mg), and phosphorus (P) levels were lower, while cardiac troponin I (cTnI), parathormone (PTH), creatine kinase (CK), and creatine kinase-MB (CK-MB) levels were higher before treatment as compared with the control group. Increases in the Ca, Mg, ICa, and P levels, and a decrease in the PTH levels were detected after treatment when compared with those levels before treatment. The CK, CK-MB, and Mg levels also increased, while the ICa level decreased in the aftertreatment group as compared with the control group. Therefore, the cTnI, CK, and CK-MB in cows with hypocalcemia were determined to be important diagnostically, but insignificant for prognosis. Homocysteine was not important for either diagnosis or prognosis.