The goal of this study was to determine the optimal location for ultrasound-guided centesis of the bovine abomasum and to assess the safety of the procedure. In the first part of this study, the technique was applied to 50 clinically healthy cows which were slaughtered within two hours of the procedure. The abomasum and peritoneum were then examined for lesions. In all but one cow, the location for abomasocentesis was 10 to 27 cm caudal to the xiphoid and on the ventral midline or up to 10 cm to the right of it. No peritoneal lesions were observed in any of the cows. In all cases, the site of centesis was visible as a localised haemorrhage on the serosal surface of the abomasum. In 41 of the cows, a haematoma was visible on the mucosal surface of the abomasum. In the second part of the study, 10 cows were monitored clinically for 10 days after abomasocentesis, to assess the safety of the procedure. The appetite, general behaviour, attitude and rectal temperature of the cows remained normal. The haematocrit, total and differential leucocyte counts, and the concentrations of total solids and fibrinogen were determined daily and remained within their normal ranges. At slaughter minimal changes, such as localised reddening and adhesions between the site of the puncture in the abomasum and the abdominal wall, were visible in three of the cows.