To investigate amniotic fluid (AF) dynamics and volume regulatory mechanisms, we measured the concentration of radioiodinated (125I) serum albumin (RISA), 51Cr-labeled red blood cells (Cr-RBC), and 103Ru-labeled microspheres after injection into the amniotic cavity and determined AF volume and fetal swallowing rate in 22 singleton pregnant sheep. Under normal conditions 2-3 h were required for complete mixing of RISA and Cr-RBC within AF; however, when the fetus was dead only 3-5 h were required. AF volume of 17 sheep on the 5th postoperative day averaged 975 +/- 128 ml by RISA and 986 +/- 130 ml by Cr-RBC. AF volume determined with RISA and Cr-RBC correlated well. In contrast, AF volume measurement with microspheres produced erratic results. The disappearance rate of the labels in 17 ewes on the 5th postoperative day averaged 4.9 +/- 0.7%/h for RISA and 5.5 +/- 0.7 for Cr-RBC, and the calculated rates of fetal swallowing were 935 +/- 78 ml/day by RISA and 1,085 +/- 102 by Cr-RBC. In dead fetuses the disappearance rates were almost zero, suggesting that the labels disappear mainly by swallowing. Absolute volume swallowed and swallowed volume per fetal weight correlated with gestational age. AF volume correlated with fetal weight. Radiolabeled albumin or red blood cells may be used to simultaneously measure amniotic fluid volume and the rate of fetal swallowing. Furthermore it appears that fetal swallowing increases with gestational age.