Abstract
We investigated methanotrophic bacteria in slightly alkaline surface water (pH 7.4–8.7) of oilsands tailings ponds in Fort McMurray, Canada. These large lakes (up to 10 km2) contain water, silt, clay and residual hydrocarbons that are not recovered in oilsands mining. They are primarily anoxic and produce methane but have an aerobic surface layer. Aerobic methane oxidation was measured in the surface water at rates up to 152 nmol CH4 ml−1 water d−1. Microbial diversity was investigated via pyrotag sequencing of amplified 16S rRNA genes, as well as by analysis of methanotroph-specific pmoA genes using both pyrosequencing and microarray analysis. The predominantly detected methanotroph in surface waters at all sampling times was an uncultured species related to the gammaproteobacterial genus Methylocaldum, although a few other methanotrophs were also detected, including Methylomonas spp. Active species were identified via 13CH4 stable isotope probing (SIP) of DNA, combined with pyrotag sequencing and shotgun metagenomic sequencing of heavy 13C-DNA. The SIP-PCR results demonstrated that the Methylocaldum and Methylomonas spp. actively consumed methane in fresh tailings pond water. Metagenomic analysis of DNA from the heavy SIP fraction verified the PCR-based results and identified additional pmoA genes not detected via PCR. The metagenome indicated that the overall methylotrophic community possessed known pathways for formaldehyde oxidation, carbon fixation and detoxification of nitrogenous compounds but appeared to possess only particulate methane monooxygenase not soluble methane monooxygenase.
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Acknowledgements
We thank the mining companies for the support of this research. Particular thanks go to Samantha Tavener and Joe Fournier for providing samples. This research was supported by Genome Canada and Genome Alberta (Hydrocarbon Metagenomics) and the Institute for Sustainable Energy, Environment and Economy (ISEEE). CES was supported by doctoral fellowships from NSERC and Alberta Innovates-Technology Futures. ALB was supported by a postdoctoral award from NSERC. DNA Sequence data sets related to this project have been submitted to the Sequence Read Archive under accession number SRP013946.
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Saidi-Mehrabad, A., He, Z., Tamas, I. et al. Methanotrophic bacteria in oilsands tailings ponds of northern Alberta. ISME J 7, 908–921 (2013). https://doi.org/10.1038/ismej.2012.163
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DOI: https://doi.org/10.1038/ismej.2012.163
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