After total hepatectomy in the dog, increased bilirubin concentrations were found in the circulating plasma or bound to tissues. Small amounts of the accumulated pigment were also excreted in the urine. Both unconjugated (UCB) and direct-reacting (DB) serum bile pigments increased significantly – especially after bilirubin loading. Analysis of the bile pigments in the serum and urine of hepatectomized dogs showed that approximately 10% of the conjugates were present as bilirubin xyloside and 25% as bilirubin glucoside. By performing an enterectomy and/or a splenectomy and also a nephrectomy, it was possible to demonstrate that the spleen, intestine, and kidney may be sites of bilirubin formation and conjugation in the hepatectomized dog.