The objective of this investigation was to validate a transit-time ultrasound blood flow metering system in vivo. Implanted chronically and acutely on the ascending aorta of the dog, the transit-time flow probe determined varying flow rates simultaneously with measurements made by the electromagnetic flow metering method. The transit-time technique was also compared to two methods in which blood was collected volumetrically by either graduated cylinder (ascending aorta/dog) or pump withdrawal (abdominal aorta/cat). Statistical analysis of the results provided evidence that the transit-time ultrasound method measured in vivo blood flow rate no differently than the electromagnetic or pump withdrawal techniques, however, transit-time determinations of blood volume were 10% below that indicated by graduated cylinder collection. With transit time represented on the y-axis, three linear regressions of all paired blood flow measurements were calculated yielding the following slopes (delta y/delta x) and regression coefficients (r), respectively: electromagnetic (1.00, 0.98), graduated cylinder (0.85, 0.93), and pump withdrawal (0.93, 1.00). The results validate the transit-time ultrasound system used in the present investigation as an accurate method capable of measuring blood flow in both acutely and chronically instrumented animal preparations.