The purpose of this study was to compare the hepatic enzyme activities of gluconeogenesis between control cows and experimental cows that had been overfed during the dry period to induce fatty liver postpartum Blood and liver samples were collected 1 wk before and 0.5, 1, 2, and 3 wk after parturition. Before parturition, neither the serum nonesterified fatty acid nor the liver triacylglycerol concentration differed between the two groups. After parturition, these variables were higher in experimental cows than in control cows. Liver glycogen was higher at 1 wk before parturition in experimental cows; sharply decreased after parturition in both groups; and, at 1 wk after parturition, was lower in experimental cows than in control cows. In the liver, activities of phosphoenolpyruvate carboxykinase were significantly lower at 1 wk before and at 0.5 and 2 wk after parturition in experimental cows; in addition, the activities tended to be lower at 1 wk after parturition. Activities of fructose 1,6-bisphosphatase tended to be lower, but activities of glucose B-phosphatase tended to be higher, at 0.5 wk after parturition in experimental cows than in control cows. Our results suggest that, in fatty infiltrated liver, the rate of gluconeogenesis is not optimal, which results in prolongation of lipolysis, particularly during the first weeks after parturition.