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Kinetic and phylogenetic characterization of an anaerobic dechlorinating microbial community

Philip Hugenholtz^2,

Jason J. Plumb^2,

¹ Water Research Institute, Via Reno 1, 00198, Rome, Italy
² Department of Microbiology and Parasitology, The University of Queensland, Brisbane, 4072, Australia
³ Department of Chemistry, University of Rome ‘La Sapienza’, P.le Aldo Moro 5, 00185 Rome, Italy

Correspondence
Simona Rossetti
rossetti{at}irsa.rm.cnr.it

The reductive dechlorination (RD) of tetrachloroethene (PCE) to vinyl chloride (VC) and, to a lesser extent, to ethene (ETH) by an anaerobic microbial community has been investigated by studying the processes and kinetics of the main physiological components of the consortium. Molecular hydrogen, produced by methanol-utilizing acetogens, was the electron donor for the PCE RD to VC and ETH without forming any appreciable amount of other chlorinated intermediates and in the near absence of methanogenic activity. The microbial community structure of the consortium was investigated by preparing a 16S rDNA clone library and by fluorescence in situ hybridization (FISH). The PCR primers used in the clone library allowed the harvest of 16S rDNA from both bacterial and archaeal members in the community. A total of 616 clones were screened by RFLP analysis of the clone inserts followed by the sequencing of RFLP group representatives and phylogenetic analysis. The clone library contained sequences mostly from hitherto undescribed bacteria. No sequences similar to those of the known RD bacteria like ‘Dehalococcoides ethenogenes’ or Dehalobacter restrictus were found in the clone library, and none of these bacteria was present in the RD consortium according to FISH. Almost all clones fell into six previously described phyla of the bacterial domain, with the majority (56·6 %) being deep-branching members of the Spirochaetes phylum. Other clones were in the Firmicutes phylum (18·5 %), the Chloroflexi phylum (16·4 %), the Bacteroidetes phylum (6·3 %), the Synergistes genus (1·1 %) and a lineage that could not be affiliated with existing phyla (1·1 %). No archaeal clones were found in the clone library. Owing to the phylogenetic novelty of the microbial community with regard to previously cultured micro-organisms, no specific microbial component(s) could be hypothetically affiliated with the RD phenotype. The predominance of Spirochaetes in the microbial consortium, the main group revealed by clone library analysis, was confirmed by FISH using a purposely developed probe.

The GenBank accession numbers for the sequences reported in this paper are AF408637–AF408652.

Present address: Department of Environmental Science, Policy and Management, Hilgard Hall, University of California Berkeley, Berkeley, CA 94720, USA.

Present address: CSIRO Land and Water, Underwood Avenue, Floreat WA 6014, Australia.

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