The effects of surgical bowel manipulation and anesthesia on intestinal glucose absorption were determined in chronically catheterized rats. Total and passive rates of glucose absorption were measured using 3-O-methyl-glucose (3OMG) and L-glucose, metabolically inert analogues of D-glucose. The rates of 3OMG absorption immediately postoperative and 4 h later were 86 and 62% less than the absorption rate 6 d postoperative. The absorption rates of 3OMG 1 and 2 d postoperative were not different from 6 d postoperative. Absorption of L-glucose was not altered by bowel manipulation and anesthesia. Even after correction for the increased resistance of the unstirred water layer (UWL) after bowel manipulation, the rates of total and active intestinal glucose absorption immediately postoperative were only 11 and 15% of predicted rates of absorption. In chronically catheterized rats, > 75% of luminal 3OMG at a concentration of 400 mM was absorbed by active transport. The Km and Vmax of 3OMG active transport corrected for the resistance of the UWL were 11.3 mM and 15.6 mumoles/min, respectively. We conclude that measurements of intestinal glucose absorption performed within 24 h of surgical bowel manipulation greatly underestimate active absorption even if corrections are made to account for the increased resistance of the UWL.