Eight mature female sheep were offered a ration which maintained body weight constant during a 20-week period. During the final 10 weeks a comparison was made in each animal of the pattern of equilibration and urinary losses of tritiated water during 8 h after dosing by four different routes. These were intravenous, intraperitoneal, intraruminal and a combination of the intraperitoneal and intraruminal routes. Tritiated water spaces were calculated from (a) the 8-h plasma specific activity and (b) by extrapolation to zero time of the plasma specific activities during the 7 days after injection. At the end of the experiment the fat and water contents of the bodies of the sheep were determined directly.

Complete equilibration of tritiated water between plasma and rumen water was not achieved in all animals 8 h after intravenous or intraperitoneal injection but was when the rumen was primed by the combination of intraperitoneal and intraruminal dosing. After intraruminal dosing equilibration was not achieved in any animal within 8 h of dosing.

Urinary losses of marker were lower after intraruminal dosing but otherwise averaged 4–5 % of the dose/1 urine. This was equivalent to 0·3–6·7% of the dose for individual sheep.

Errors resulting from incomplete equilibration and urinary loss of marker did not influence the efficiency of prediction of total body water from tritiated water space. The multiple correlation coefficient relating body fat with empty body weight and its water content was very high (r = 0·99). Errors introduced into this relationship by the inclusion of gut water in the prediction equations were apparently of a similar magnitude to those resulting from the errors in the estimation of tritiated water space.

The extrapolation method for the determination of tritiated water space was shown to have the same accuracy as equilibration techniques under these controlled dietary conditions.