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Applied and Environmental Microbiology, February 2005, p. 1058-1065, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.1058-1065.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

Desulfitobacterium hafniense Is Present in a High Proportion within the Biofilms of a High-Performance Pentachlorophenol-Degrading, Methanogenic Fixed-Film Reactor

M. Lanthier,^1, P. Juteau,¹ F. Lépine,¹ R. Beaudet,¹ and R. Villemur^1*

INRS—Institut Armand-Frappier, Laval, Québec, Canada¹

Received 2 July 2004/ Accepted 21 September 2004

We developed a pentachlorophenol (PCP)-degrading, methanogenic fixed-film reactor by using broken granular sludge from an upflow anaerobic sludge blanket reactor. This methanogenic consortium was acclimated with increasing concentrations of PCP. After 225 days of acclimation, the reactor was performing at a high level, with a PCP removal rate of 1,173 µM day^–1, a PCP removal efficiency of up to 99%, a degradation efficiency of approximately 60%, and 3-chlorophenol as the main chlorophenol residual intermediate. Analyses by PCR-denaturing gradient gel electrophoresis (DGGE) showed that Bacteria and Archaea in the reactor stabilized in the biofilms after 56 days of operation. Important modifications in the profiles of Bacteria between the original granular sludge and the reactor occurred, as less than one-third of the sludge DGGE bands were still present in the reactor. Fluorescence in situ hybridization experiments with probes for Archaea or Bacteria revealed that the biofilms were composed mostly of Bacteria, which accounted for 70% of the cells. With PCR species-specific primers, the presence of the halorespiring bacterium Desulfitobacterium hafniense in the biofilm was detected very early during the reactor acclimation period. D. hafniense cells were scattered in the biofilm and accounted for 19% of the community. These results suggest that the presence of PCP-dehalogenating D. hafniense in the biofilm was crucial for the performance of the reactor.

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* Corresponding author. Mailing address: INRS—Institut Armand-Frappier, 531 Boul. des Prairies, Laval, Québec, Canada H7V 1B7. Phone: (450) 687-5010, ext. 4611. Fax: (450) 686-5501. E-mail: richard.villemur{at}inrs-iaf.uquebec.ca.

Present address: Department of Microbiology, Morrill Science Center IV North, University of Massachusetts, Amherst, MA 01003-9298.

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Applied and Environmental Microbiology, February 2005, p. 1058-1065, Vol. 71, No. 2
0099-2240/05/$08.00+0     doi:10.1128/AEM.71.2.1058-1065.2005
Copyright © 2005, American Society for Microbiology. All Rights Reserved.

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