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

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.