Deconjugation of bilirubin conjugates and urobilin formation by conventionalized germ-free rats

The amounts of conjugated bilirubin and urobilins/urobilinogen were determined semiquantitatively in faeces of germ-free (GF) rats during GF condition and after conventionalization by oral administration of faeces suspension from conventional (CONV) rats. The amount of bilirubin conjugates, detected as their ethyl anthranilate azopigments, decreased rapidly 1 day after conventionalization. Thin-layer chromatography analysis of the corresponding faecal azopigment preparations showed that some azopigments started to disappear a few days after the conventionalization, indicating that their corresponding bilirubin conjugates were deconjugated by the bacteria in the intestine. On day 21 after conventionalization, only two azopigments were detected, namely the unconjugated and glucuronic acid conjugated dipyrroles of bilirubin, respectively, thus indicating the presence of only one bilirubin conjugate, the monoglucuronide. After 69 days no azopigments could be detected, indicating the total absence of conjugated bilirubin in these faeces samples. No urobilins were detected in faeces of the rats during their GF state, but these metabolites appeared in faeces one day after conventionalization and increased during a few days to a CONV level.