Isolation and Purification of the Oxalate-Degrading Bacterium Oxalobacter formigenes in Human Feces.
- Author:
Cheol KWAK
1
;
Ji Eun OH
;
Hee Kyung KIM
;
Myung Sik CHOI
;
Eui Chong KIM
;
Hyeon Hoe KIM
Author Information
1. Department of Urology, Seoul National University College of Medicine and Clinical Research Institute, Seoul National University Hospital, Seoul, Korea. hhkim@snu.ac.kr
- Publication Type:Original Article
- Keywords:
Kidney calculi;
Oxalobacter formigenes;
Isolation & Purification
- MeSH:
Bacteria;
Calcium Oxalate;
DNA;
Feces*;
Gastrointestinal Tract;
Humans*;
Kidney Calculi;
Oxalobacter formigenes*;
Polymerase Chain Reaction;
Rumen;
Sheep;
Urolithiasis;
Volunteers
- From:Korean Journal of Urology
2003;44(8):785-790
- CountryRepublic of Korea
- Language:Korean
-
Abstract:
PURPOSE: The absence of the oxalate-degrading bacteria, Oxalobacter formigenes, in the gastrointestinal tract correlates with the formation of calcium-oxalate urolithiasis. The aim of this study was to isolate Oxalobacter from human feces. MATERIALS AND METHODS: The OxB strain, isolated from sheep rumen, was incubated in selective growth media(medium B) in an anaerobic chamber, and its microbiological properties evaluated. Feces from volunteers, who were presumed to have O. formigenes from a polymerase chain reaction-based detection system, was incubated in medium B. The colonies isolated, primarily 7 days after incubation, were successively subcultured, and colony-PCR performed to isolate colonies from the successive subcultures. RESULTS: The colonies of OxB were Gram-negative, non-motile, non-spore forming, rod-shaped cells. The cells were often in pairs or chains. OxB DNA gave rise to an amplicon of the correct molecular size(416 bp) of O. formigenes. The morphology of the colonies from human feces, of which DNA was confirmed to have the same size amplicon of O. formigenes by PCR, was identical to the OxB, both grossly and by Gram stain. Although the morphology of the colonies isolated from the successive subcultures was no different from that of OxB, the PCR positivity of the isolated colonies decreased on successive subculturing, with no PCR-positive colonies from the fifth subculture. CONCLUSIONS: Our results suggest that the microbiological isolation and purification of Oxalobacter formigenes from human feces is a difficult procedure. Special culture conditions will be required to culture Oxalobacter species to reveal the link between O. formigenes and calcium oxalate urolithiasis in humans.
