A H2-based, denitrifying membrane-biofilm reactor (MBfR) was effective for removing trichloroethene (TCE) by reductive dechlorination. When TCE was first added to the MBfR, reductive dechlorination took place immediately and then increased over 18 weeks, and TCE was completely dechlorinated to ethene by about 120 days. These results indicate that TCE-dechlorinating bacteria were present naturally in the H2-based biofilm, and that enrichment for TCE-dechlorinating bacteria occurred. Dehalococcoides were documented in the MBfR biofilm before and after TCE feeding. Their proportion, quantified using the 16S rRNA gene, increased from 2.9 to 12% after TCE addition. This is the first report in which Dehalococcoides are proven to be present as part of an autotrophic biofilm community active in reductive dechlorination of TCE to ethene in a laboratory controlled experiment. Based on the complete reduction of TCE to ethene, the 16S rRNA clone libraries results, and the amount of tceA and bvcA, it appears that at least two Dehalococcoides strains were present in the enriched biofilm. One of them seems to be a new strain that is unique for having tceA and bvcA reductive dehalogenases.