The metabolic and physiologic responses to 7-hour endotoxin infusion (5.0 mg/kg h) were evaluated in guinea pigs following 6 weeks of dietary enrichment with diets containing either chemically structured lipid (SL) composed of medium-chain triglycerides (MCT) and long-chain triglycerides (LCT) in the form of N-3 polyunsaturated fatty acids (PUFAs), or safflower oil (SO), which is high in N-6 fatty acids. Plasma phospholipid fatty acid profiles, arterial blood pH, PCO2, PO2, HCO3, lactate, blood pressure, oxygen consumption, and energy expenditure were examined. Plasma phospholipid fatty acids profiles reflected dietary intake with SL-fed animals demonstrating a significantly higher N-3 to N-6 fatty acid ratio compared with SO-fed animals. SL-fed animals responded to endotoxemia with a mild metabolic acidosis with respiratory compensation, which was associated with moderate lactatemia (3 mmol/L). SO-fed animals developed a severe metabolic acidosis with acidemia and respiratory compensation, which was associated with hyperlactatemia (8 mmol/L, P less than .05 v SL). No differences were observed in blood pressure, oxygen consumption, energy expenditure, or respiratory quotient during endotoxemia between dietary groups compared with controls. We conclude that diets enriched with structured lipid composed of medium-chain and N-3 fatty acids can attenuate the sequelae of endotoxemia.