Isolation and characterization of cholic acid 7alpha-dehydroxylating fecal bacteria from cholesterol gallstone patients

J Hepatol. 2000 Jan;32(1):4-10. doi: 10.1016/s0168-8278(00)80183-x.

Abstract

Background/aims: The development of cholesterol gallstones, in some patients, has been associated with increased proportions of deoxycholic acid in the bile acid pool. Deoxycholic acid is a microbial product of cholic acid 7alpha-dehydroxylation in the intestines. The levels and activities of bile acid 7alpha-dehydroxylating bacteria have been reported to be increased in gallstone patients. The aim of the current study was to isolate 7alpha-dehydroxylating bacteria from gallstone patients and determine if these individuals are colonized by similar bacterial species.

Methods: The levels of 7alpha-dehydroxylating bacteria in fecal samples were determined by fecal dilutions in 24 gallstone patients and 10 controls. 7alpha-Dehydroxylating bacteria were isolated by a non-selective streak plate technique and 7alpha-dehydroxylation activity was determined by measuring the conversion of [14C]-cholic acid to [14C]-deoxycholic acid using thin-layer chromatography.

Results: Gallstone patients had >42-fold (p<0.01) higher levels of 7alpha-dehydroxylating bacteria than patients who had not developed gallstones. Eighteen strains of 7alpha-dehydroxylating bacteria were isolated from eight gallstone patients. Attempts to isolate 7alpha-dehydroxylating bacteria from ten control patients were unsuccessful using identical isolation techniques. Surprisingly, all strains of bacteria isolated from gallstone patients appear to belong to the genus Clostridium.

Conclusion: Gallstone patients have higher levels of 7alpha-dehydroxylating fecal bacteria and appear to harbor only members of the genus Clostridium with this activity.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Adult
  • Aged
  • Bacterial Typing Techniques
  • Blotting, Southern
  • Cholelithiasis / microbiology*
  • Cholesterol*
  • Cholic Acid / metabolism
  • Cholic Acids / metabolism*
  • Chromatography, Thin Layer
  • Clostridium / genetics
  • Clostridium / isolation & purification
  • Clostridium / metabolism*
  • Colony Count, Microbial
  • DNA, Bacterial / analysis
  • Deoxycholic Acid / metabolism
  • Eubacterium / genetics
  • Eubacterium / isolation & purification
  • Eubacterium / metabolism*
  • Feces / microbiology*
  • Female
  • Humans
  • Male
  • Middle Aged
  • Steroid Hydroxylases / metabolism*

Substances

  • Cholic Acids
  • DNA, Bacterial
  • Deoxycholic Acid
  • Cholesterol
  • Steroid Hydroxylases
  • Cholic Acid