Combined anaerobic ammonium and methane oxidation for nitrogen and methane removal

et al. 2015

Appl Environ Microbiol

Anaerobic oxidation of methane (AOM) coupled to nitrite reduction is a novel AOM process that is mediated by denitrifying methanotrophs. To date, enrichments of these denitrifying methanotrophs have been confined to freshwater systems; however, the recent findings of 16S rRNA and pmoA gene sequences in marine sediments suggest a possible occurrence of AOM coupled to nitrite reduction in marine systems. In this research, a marine denitrifying methanotrophic culture was obtained after 20 months of enrichment. Activity testing and quantitative PCR (qPCR) analysis were then conducted and showed that the methane oxidation activity and the number of NC10 bacteria increased correlatively during the enrichment period. 16S rRNA gene sequencing indicated that only bacteria in group A of the NC10 phylum were enriched and responsible for the resulting methane oxidation activity, although a diverse community of NC10 bacteria was harbored in the inoculum. Fluorescence in situ hybridization showed that NC10 bacteria were dominant in the enrichment culture after 20 months. The effect of salinity on the marine denitrifying methanotrophic culture was investigated, and the apparent optimal salinity was 20.5‰, which suggested that halophilic bacterial AOM coupled to nitrite reduction was obtained. Moreover, the apparent substrate affinity coefficients of the halophilic denitrifying methanotrophs were determined to be 9.8 ؎ 2.2 M for methane and 8.7 ؎ 1.5 M for nitrite.A naerobic oxidation of methane (AOM) occurs extensively in natural ecosystems and is a crucial biological sink in the global methane cycle that maintains the balance of greenhouse gas content in the atmosphere (1). To date, five electron acceptors that support AOM, including AOM coupled to sulfate reduction (2), nitrite reduction (3), nitrate reduction (4), iron reduction (5), and manganese reduction (5), have been discovered in natural settings. Moreover, on the basis of bioenergetic calculations, researchers have speculated that several other types of AOM (e.g., AOM coupled to perchlorate reduction, arsenate reduction, and selenate reduction) may exist in nature (6); however, these possible types have not yet been confirmed. In particular, AOM coupled to nitrite reduction has been a predominant research focus in the past several years. AOM coupled to nitrite reduction was also called nitrite-dependent anaerobic methane oxidation (ndamo) in previous reports. It has been demonstrated that AOM coupled to nitrite reduction is mediated by the bacterium "Candidatus Methylomirabilis oxyfera" (denitrifying methanotroph) (7), which is affiliated with the candidate NC10 phylum (8). This candidate division (NC10 phylum) was first defined by classification of environmental sequences retrieved from aquatic microbial formations in flooded caves (9). To date, our knowledge regarding the NC10 phylum has stemmed largely from research into "Ca. Methylomirabilis oxyfera" and "Ca. Methylomirabilis oxyfera"-like bacteria. According to the phylogenetic affiliations of 16S rRNA gene sequ...

Section: Figsupporting

confidence: 55%

Anaerobic Oxidation of Methane Coupled to Nitrite Reduction by Halophilic Marine NC10 Bacteria

Geng

et al. 2015

Appl Environ Microbiol

“…S2), which was from the inoculum (Table S1) and the decomposition of organic matters. Since anaerobic ammonia oxidation (anammox) bacteria have a higher affinity for nitrite than n-damo bacteria (Luesken et al 2011a;Zhu et al 2011), anammox reaction might be significant in phase I, especially in the first 50 days. In short, n-damo bacteria had intense competitions from denitrifying and anammox bacteria when methanogenic sludge was selected as inoculum.…”

Section: Discussionmentioning

confidence: 99%

Effect of inoculum sources on the enrichment of nitrite-dependent anaerobic methane-oxidizing bacteria

Appl Microbiol Biotechnol

Shen

et al. 2014

Nitrite-dependent anaerobic methane oxidation (n-damo) is a newly discovered biological process that couples anaerobic oxidation of methane (AOM) to nitrite reduction. In this study, three different inocula, methanogenic sludge, paddy soil, and freshwater sediment were used to enrich n-damo bacteria in three sequencing batch reactors (SBRs), and three n-damo enrichment cultures, C1, C2 and C3, were obtained, respectively. After 500 days of incubation, Methylomirabilis oxyfera-like bacteria and n-damo activities were observed in cultures C1, C2, and C3, and the specific activities were 0.8 ± 0.1, 1.4 ± 0.1, and 1.0 ± 0.1 μmol CH4 h(-1) g(-1) VSS, respectively. The copy numbers of 16S rRNA genes from cultures C1, C2, and C3 were 5.0 ± 0.4 × 10(8), 6.1 ± 0.1 × 10(9), and 1.0 ± 0.2 × 10(9) copies g(-1) dry weight, respectively. The results indicated that paddy soil is an excellent inoculum for n-damo bacterial enrichment. This work expanded the alternative source of n-damo inoculum and benefited the further research of n-damo process.

“…1), and these NC10 bacteria were physiologically active as denitrifying methanotrophs. Several enrichment cultures have indicated that group A members of the NC10 phylum perform AOM coupled to nitrite reduction45678910. Remarkably, the typical bacterium of the NC10 phylum, ‘ Candidatus Methylomirabilis oxyfera’ ( M. oxyfera ), utilizes oxygen produced from nitric oxide to intra-aerobically oxidize methane under anoxic conditions11.…”

mentioning

confidence: 99%

A novel denitrifying methanotroph of the NC10 phylum and its microcolony

Wang

et al. 2016

Sci Rep

The NC10 phylum is a candidate phylum of prokaryotes and is considered important in biogeochemical cycles and evolutionary history. NC10 members are as-yet-uncultured and are difficult to enrich, and our knowledge regarding this phylum is largely limited to the first species ‘Candidatus Methylomirabilis oxyfera’ (M. oxyfera). Here, we enriched NC10 members from paddy soil and obtained a novel species of the NC10 phylum that mediates the anaerobic oxidation of methane (AOM) coupled to nitrite reduction. By comparing the new 16S rRNA gene sequences with those already in the database, this new species was found to be widely distributed in various habitats in China. Therefore, we tentatively named it ‘Candidatus Methylomirabilis sinica’ (M. sinica). Cells of M. sinica are roughly coccus-shaped (0.7–1.2 μm), distinct from M. oxyfera (rod-shaped; 0.25–0.5 × 0.8–1.1 μm). Notably, microscopic inspections revealed that M. sinica grew in honeycomb-shaped microcolonies, which was the first discovery of microcolony of the NC10 phylum. This finding opens the possibility to isolate NC10 members using microcolony-dependent isolation strategies.