We used in situ hybridization with fluorescently labeled rRNA-targeted oligonucleotide probes concurrently with microscopic examinations and methane measurements to characterize the microbial community of an anaerobic hybrid reactor treating pentachlorophenol (PCP) with a mixture of fatty acids (propionic, butyric, acetic and lactic) and methanol. Archaeal cells detected with probe ARC915 prevailed in anaerobic granular sludge without and with the addition of PCP in a range of 2.0 to 21.0 mg/L to the reactor. This group accounted for 81 and 90% of the DAPI-stained cells before and after the addition of 21 mg/L of PCP, respectively. In these conditions, cells detected with the Methanosarcinales specific probe (MSMX860) were the only methanogenic Archaea found and accounted for 59 to 87.6% of the DAPI-stained cells. No cells were detected by the Methanomicrobiales (MG1200), Methanobacteriaceae (MB1174) and Methanococcaceae (MC1109) specific probes. Bacterial cells detected with probe EUB338 were found in very low numbers, which ranged from 5.7 to 1.0% of the DAPI-stained cells. This finding agrees with the scanning electron microscope examinations, in which cells morphologically resembling Methanosaeta and Methanosarcina were predominantly observed in the granular sludge. Results contributed to the investigation of the importance of the methanogens during PCP degradation.

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