AbstractModern, molecular microbiological methods were applied to urine samples from control subjects and athletes for characterization of the microbial community. High abundance of lactobacilli, enterococci, and enterobacteria was detected in urine samples, suggesting that gastrointestinal and urogenital tracts act as contamination sources. Athlete samples, but not control samples, showed an abundance of pseudomonads, a bacterial group reported to metabolize steroids. Overall, the bacteria detected are known to be capable of altering steroid profiles, emphasizing the importance of good hygiene at sampling in reliable doping control. Copyright © 2010 John Wiley & Sons, Ltd.