In the past, hepatic blood flow in cows was invasively characterized to investigate different pathologies and physiological conditions. However, hepatic blood flow can be easily evaluated with transabdominal Doppler ultrasound. Sixteen healthy adult non-lactating, non-pregnant Holstein-Friesian cows were examined using B-mode and Doppler ultrasound between the right flank and 9th intercostal space to establish the best approach to the different parts of the portal and hepatic vein systems, and determine normal blood flow characteristics. The main portal vein was characterized by a turbulent, high-velocity flow due to the opposing confluence of the splenic and cranial mesenteric veins, while hepatic and caudal vena cava veins have laminar blood flow, in which the phasicity is considered mainly respiratory in origin. Reference values were determined in relation to the anatomical point of observation. In conclusion, transabdominal Doppler ultrasound of the portal system is a simple technique that allows non-invasive characterization of portal and hepatic blood haemodynamics in cows.