Twenty-four high producing, multiparous Holstein cows were utilized to evaluate effects of long-term administration of sustained-release rbST on blood pH, gases, buffer capacity, circulating metabolites and hormones, and on liver lipid content and functions during lactation. Treatment, commencing 98 to 112 d postpartum and continued until d 305, consisted of a subcutaneous injection of a placebo or 350 mg of rbST every 14 d. Milk and 3.5% FCM were increased significantly by rbST, but milk fat and protein content, feed intake, energy balance, corrected feed efficiency for milk production, and body condition scores were unaffected. Somatotropin decreased blood pH and buffer capacity by decreasing bicarbonate without affecting blood partial pressures of oxygen or carbon dioxide. Concentration of plasma albumin was decreased and nonesterified fatty acids, glucose, insulin, rbST and insulin-like growth factor-I were increased by treatment. Total plasma proteins and cortisol, hemoglobin, and hematocrit were unaffected. Liver DM, total lipid and triacylglycerol contents, and plasma 3-hydroxybutyrate and glutamic oxalacetic transaminase were not affected by rbST, but the percentage of triacylglycerol in total lipids was increased. Results of this experiment suggest that rbST exhibited lipolytic and diabetogenic activities in lactating dairy cows in vivo and that these effects might be important for the increase in nutrient partitioning toward the mammary gland elicited by rbST.