No method exists which enables easy, frequent, and, at the same time, reliable cardiac output (CO) measurements in mice. To validate a simple indicator-dilution method suitable for frequent measurements of CO in small laboratory animals, a 5% glucose solution was injected as a bolus into femoral veins of mice and rats. The corresponding blood conductivity was measured continuously between an intra-aortic and a rectal electrode. The resulting conductivity-dilution curves were used to calculate CO in mice during hypervolemia and hypovolemia and in conscious as well as halothane-anesthetized mice and rats. In rats, conductivity-dilution curves and time courses of plasma glucose concentration were recorded simultaneously. Compared with CO in awake animals, CO in both species was slightly, but not significantly, reduced during halothane anesthesia. CO was significantly and gradually reduced in hypovolemic mice (up to 58 ml blood/kg body wt), whereas hypervolemia (23 ml saline/kg body wt) had no significant effect. Simultaneous recordings of conductivity-dilution curves and time courses of plasma glucose concentration yielded corresponding values of CO (P < 0.001). Measurement of blood conductivity appears to be a reliable, simple, and convenient method for quantification of CO in small animals.