A synthetic wastewater containing phenol as sole substrate was treated in a 2.8 L upflow anaerobic sludge blanket (UASB) reactor at ambient temperature. The operation conditions and phenol removal efficiency were discussed, microbial population in the UASB sludge was identified based on DNA cloning, and pathway of anaerobic phenol degradation was proposed. Phenol in wastewater was degraded in an UASB reactor at loading rate up to 18 gCOD/(L x d), with a 1:1 recycle ratio, at 26 +/- 1 degrees C, pH 7.0-7.5. An UASB reactor was able to remove 99% of phenol up to 1226 mg/L in wastewater with 24 h of hydraulic retention time (HRT). For HRT below 24 h, phenol degradation efficiency decreased with HRT, from 95.4% at 16 h to 93.8% at 12 h. It further deteriorated to 88.5% when HRT reached 8 h. When the concentration of influent phenol of the reactor was 1260 mg/L (corresponding COD 3000 mg/L), with the HRT decreasing (from 40 h to 4 h, corresponding COD loading increasing), the biomass yields tended to increase from 0.265 to 3.08 g/(L x d). While at 12 h of HRT, the biomass yield was lower. When HRT was 12 h, the methane yield was 0.308 L/(gCOD removed), which was the highest. Throughout the study, phenol was the sole organic substrate. The effluent contained only residual phenol without any detectable intermediates, such as benzoate, 4-hydrobenzoate or volatile fatty acids (VFAs). Based on DNA cloning analysis, the sludge was composed of five groups of microorganisms. Desulfotomaculum and Clostridium were likely responsible for the conversion of phenol to benzoate, which was further degraded by Syntrophus to acetate and H2/CO2. Methanogens lastly converted acetate and H2/CO2 to methane. The role of epsilon-Proteobacteria was, however, unsure.