Kyung-Eun Moon

Earthworm cast as a promising filter bed material and its methanotrophic contribution to methane removal

The use of biocovers is a promising strategy toward mitigating CH(4) emission from smaller and/or older landfills. In this study, a filter bed material consisting of a mixture of earthworm cast and rice paddy soil in a biocover was evaluated. Although the CH(4) oxidation rate of the enriched paddy soil was 4.9 microg g-dry soil(-1) h(-1), it was enhanced to 25.1 microg g-dry soil(-1) h(-1) by adding an earthworm cast with a 3:7 ratio of earthworm cast:soil (wet weight). CO(2) was found as the final oxidation product of CH(4), and the mole ratio of CO(2) production to CH(4) consumption was 0.27. At a moisture content range of 15-40% and a temperature range of 20-40 degrees C, the CH(4) oxidation rates of the enriched mixture were more than 57% of the maximum rate obtained at 25% moisture content and 25 degrees C. By denaturing gradient gel electrophoresis analysis employing primers for the universal bacterial 16S rRNA gene, and terminal-restriction fragment length polymorphism analysis using primers for the pmoA gene, the bacterial and methanotrophic communities in the enriched mixture were mainly originate from paddy soil and earthworm cast, respectively. Both type I (mainly Methylocaldum) and type II methanotrophs (mainly Methylocystis) played important roles in CH(4) oxidation in the enriched mixture.

Depth profiles of methane oxidation potentials and methanotrophic community in a lab-scale biocover

The depth profiles of the CH4 oxidation potentials and the methanotrophic community were characterized in a lab-scale soil mixture biocover. The soil mixture samples were collected from the top (0-10cm), middle (10-40cm), and bottom (40-50cm) layers of the biocover where most of methane was oxidized at the top layer due to consumption of O2. Batch tests using serum bottles showed that the middle and bottom samples displayed CH4 oxidation activity under aerobic conditions, and their CH4 oxidation rates were 85 and 71% of the rate of top sample (8.40μmolgdry sample(-1)h(-1)), respectively. The numbers of methanotrophs in the middle and bottom were not significantly different from those in the top sample. There was no statistical difference in the community stability indices (diversity and evenness) among the methanotrophic communities of the three layer samples, even though the community structures were distinguished from each other. Based on microarray analysis, type I and type II methanotrophs were equally present in the top sample, while type I was more dominant than type II in the middle and bottom samples. We suggested that the qualitative difference in the community structures was probably caused by the difference in the depth profiles of the CH4 and O2 concentrations. The results for the CH4 oxidation potential, methanotrophic biomass, and community stability indices in the middle and bottom layer samples indicated that the deeper layer in the methanotrophic biocover serves as a bioresource reservoir for sustainable CH4 mitigation.

Long-term performance and bacterial community dynamics in biocovers for mitigating methane and malodorous gases

The long-term performance of lab-scale biocovers for the simulation of engineered landfill cover soils was evaluated. Methane (CH4), trimethylamine (TMA), and dimethyl sulfide (DMS) were introduced into the biocovers as landfill gases for 134 days and the removal performance was evaluated. The biocover systems were capable of simultaneously removing methane, TMA, and DMS. Methane was mostly eliminated in the top layer of the systems, while TMA and DMS were removed in the bottom layer. Overall, the methane removal capacity and efficiency were 224.8±55.6g-CH4m-2d-1 and 66.6±12.8%, respectively, whereas 100% removal efficiencies of both TMA and DMS were achieved. Using quantitative PCR and pyrosequencing assay, the bacterial and methanotrophic communities in the top and bottom layers were analyzed along with the removal performance of landfill gases in the biocovers. The top and bottom soil layers possessed distinct communities from the original inoculum, but their structure dynamics were different from each other. While the structures of the bacterial and methanotrophic communities showed little change in the top layer, both communities in the bottom layer were considerably shifted by adding TMA and DMA. These findings provide information that can extend the understanding of full-scale biocover performance in landfills.

Inhibitory effects of sulfur compounds on methane oxidation by a methane-oxidizing consortium

Kinetic and enzymatic inhibition experiments were performed to investigate the effects of methanethiol (MT) and hydrogen sulfide (H2S) on methane oxidation by a methane-oxidizing consortium. In the coexistence of MT and H2S, the oxidation of methane was delayed until MT and H2S were completely degraded. MT and H2S could be degraded, both with and without methane. The kinetic analysis revealed that the methane-oxidizing consortium showed a maximum methane oxidation rate (Vmax) of 3.7 mmol g-dry cell weight (DCW)(-1) h(-1) and a saturation constant (Km) of 184.1 μM. MT and H2S show competitive inhibition on methane oxidation, with inhibition values (Ki) of 1504.8 and 359.8 μM, respectively. MT was primary removed by particulate methane monooxygenases (pMMO) of the consortium, while H2S was degraded by the other microorganisms or enzymes in the consortium. DNA and mRNA transcript levels of the pmoA gene expressions were decreased to ∼10(6) and 10(3)pmoA gene copy number g-DCW(-1) after MT and H2S degradation, respectively; however, both the amount of the DNA and mRNA transcript recovered their initial levels of ∼10(7) and 10(5)pmoA gene copy number g-DCW(-1) after methane oxidation, respectively. The gene expression results indicate that the pmoA gene could be rapidly reproducible after methane oxidation. This study provides comprehensive information of kinetic interactions between methane and sulfur compounds.