In the present study, anaerobic biotransformation of fluorene and phenanthrene by sulfate-reducing bacteria (SRB) was investigated and biotransformation pathways were proposed. SRB was enriched from anaerobic swine wastewater sludge and its abundance was determined by the fluorescence in situ hybridization (FISH) technique. Batch anaerobic biotransformation studies were conducted with fluorene (5 mg L-1), phenanthrene (5 mg L-1) and a mixture of the two (10 mg L-1). After 21 d of incubation. 88% of fluorene and 65% of phenanthrene were biotransformed by SRB. In contrast to previous studies, a decrease in biotransformation efficiency was observed in the presence of both fluorene and phenanthrene. Throughout the study, sulfate reduction was coupled with biotransformation of fluorene and phenanthrene. However, no increase in bacterial cell density was observed in the presence of an inhibitor, i.e. molybdate. Identification of metabolites by gas chromatography-mass spectrometry (GC-MS) revealed that fluorene and phenanthrene were biotransformed through a sequence of hydration and hydrolysis reactions followed by decarboxylation with the formation of p-cresol (only in the phenanthrene system) and phenol. The metabolites identified suggest novel biotransformation pathways of fluorene and phenenthrene. (C) 2008 Elsevier B.V. All rights reserved.