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Molecular characterisation of bacteria in a wetland used to remove ammoniacal-N from landfill leachate

School of Health and Bioscience, University of East London, Romford Road, London, E15 4LZ

Tom C.J. Moffett

School of Health and Bioscience, University of East London, Romford Road, London, E15 4LZ.

Bruce F. Moffett

School of Health and Bioscience, University of East London, Romford Road, London, E15 4LZ

James A. Harris

Institute of Water and Environment, Cranfield University, Silsoe, Bedfordshire. MK45 4DT

Phil J. Shaw

Institute of Water and Environment, Cranfield University, Silsoe, Bedfordshire. MK45 4DT

Joanne S. Wallace

DEFRA, Cromwell House, London. SW1P 3JH

Bacteria associated with leachate from a constructed, surface-flow wetland effective at removing ammonium-N, were characterised using molecular methods which bypass the need for cultivation. DNA was extracted from the sample and bacterial 16S rDNA sequences amplified and cloned. Ninety-six individual clones were re-amplified and analysed by restriction digestion and targeted sequencing. A collector's curve was used to estimate the total number of bacterial groups (operational taxonomic units, OTUs) in the sample at 47. Thirty-six clones representing 28 OTUs were sequenced and characterised by aligning them against published sequences. Of these, 10 were confidently identified, whilst for the remaining 18, the closest match was obtained. The sequences in 64% of the OTUs were similar to those of the Proteobacteria but no classical nitrifying bacteria were identified. However, alkaliphilic Methylomicrobium sp. that oxidise ammonium and bacteria from the genus Alcaligenes, at least one species of which is capable of heterotrophic nitrification, were detected.

Key Words: 16S rDNA • wetland • ARDRA • sequencing • passive treatment • landfill leachate • wmr 503-6.

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