In Situ Enhancement and Isotopic Labeling of Biogenic Coalbed Methane.

Subsurface microbial (biogenic) methane production is an important part of the global carbon cycle that has resulted in natural gas accumulations in many coal beds worldwide. Laboratory studies suggest that complex carbon-containing nutrients (e.g., yeast or algae extract) can stimulate methane production, yet the effectiveness of these nutrients within coal beds is unknown. Here, we use downhole monitoring methods in combination with deuterated water (D2O) and a 200-liter injection of 0.1% yeast extract (YE) to stimulate and isotopically label newly generated methane. A total dissolved gas pressure sensor enabled real-time gas measurements (641 days preinjection and for 478 days postinjection). Downhole samples, collected with subsurface environmental samplers, indicate that methane increased 132% above preinjection levels based on isotopic labeling from D2O, 108% based on pressure readings, and 183% based on methane measurements 266 days postinjection. Demonstrating that YE enhances biogenic coalbed methane production in situ using multiple novel measurement methods has immediate implications for other field-scale biogenic methane investigations, including in situ methods to detect and track microbial activities related to the methanogenic turnover of recalcitrant carbon in the subsurface.

Geochemical and Geophysical Monitoring of Hydrocarbon Seepage in the Adriatic Sea.

Hydrocarbon seepage is overlooked in the marine environment, mostly due to the lack of high-resolution exploration data. This contribution is about the set-up of a relocatable and cost-effective monitoring system, which was tested on two seepages in the Central Adriatic Sea. The two case studies are an oil spill at a water depth of 10 m and scattered biogenic methane seeps at a water depth of 84 m. Gas plumes in the water column were detected with a multibeam system, tightened to sub-seafloor seismic reflection data. Dissolved benthic fluxes of nutrients, metals and Dissolved Inorganic Carbon (DIC) were measured by in situ deployment of a benthic chamber, which was used also for the first time to collect water samples for hydrocarbons characterization. In addition, the concentration of polycyclic aromatic hydrocarbons, as well as major and trace elements were analyzed to provide an estimate of hydrocarbon contamination in the surrounding sediment and to make further inferences on the petroleum system.

Potential of biogenic methane for pilot-scale fermentation ex situ with lump anthracite and the changes of methanogenic consortia.

Pilot-scale fermentation is one of the important processes for achieving industrialization of biogenic coalbed methane (CBM), although the mechanism of biogenic CBM remains unknown. In this study, 16 samples of formation water from CBM production wells were collected and enriched for methane production, and the methane content was between 3.1 and 21.4%. The formation water of maximum methane production was used as inoculum source for pilot-scale fermentation. The maximum methane yield of the pilot-scale fermentation with lump anthracite amendment reached 13.66 µmol CH4/mL, suggesting that indigenous microorganisms from formation water degraded coal to produce methane. Illumina high-throughput sequencing analysis revealed that the bacterial and archaeal communities in the formation water sample differed greatly from the methanogic water enrichment culture. The hydrogenotrophic methanogen Methanocalculus dominated the formation water. Acetoclastic methanogens, from the order Methanosarcinales, dominated coal bioconversion. Thus, the biogenic methanogenic pathway ex situ cannot be simply identified according to methanogenic archaea in the original inoculum. Importantly, this study was the first time to successfully simulate methanogenesis in large-capacity fermentors (160 L) with lump anthracite amendment, and the result was also a realistic case for methane generation in pilot-scale ex situ.

The characteristics of coalbed water and coal in a coal seam situated in the Red River Basin, Vietnam.

An in-depth understanding of the hydrogeochemical characteristics of coal mines is helpful in establishing an effective and successful exploration program of coalbed methane (CBM). This study provides a comprehensive analysis of hydrogeological characteristics, characteristics of coalbed water, and characteristics of the coal sample from a coal seam located in the Red River Basin (RRB). These physicochemical characteristics along with the microbial composition of coalbed water were critically analyzed. A high concentration of chloride and sodium was found in the coalbed water, presumably due to the coal mine's stratigraphic association with marine or marine-transitional beds. A correlation between the occurrence of microbes and the chemical components in the coalbed water was established. The characteristics of the coal were systematically analyzed, including proximate, ultimate, and petrographic analyses. Based on the coal macerals, coal rank is classified as low-rank (sub-bituminous) with a vitrinite reflectance (Ro, max) of 0.36%, suggesting that this type of low-rank coal is favorable for biogenic methane generation. Pore structures and pore types were characterized using different methods, including low-temperature nitrogen adsorption/desorption (LTNA), mercury intrusion porosimetry (MIP), and scanning electron microscopy (SEM). Coal from the study area has microporous and macroporous features. Pore types of the coal were also characterized using SEM. The primary genetic pore types of the Red River coal include plant tissue holes and blowholes.

Accelerated Bioconversion of Chemically Solubilized Lignite Solution to Methane by Methanogenic Consortium: Experimental Results and Their Application to the Subsurface Cultivation and Gasification Method.

Lignite is an obsolete and less commercially circulated natural resource due to its low calorific value worldwide. The effective conversion of lignite into methane is important considering the global energy crunch. This study reported the effective bioconversion of organic matter released from chemically solubilized lignite to methane using two methanogenic consortia types: mixed methanogenic enrichment culture (mMEC) and SAL25-2. We demonstrated in a microcosm study that the start of methane generation was observed within seven days. Furthermore, the methane yield increased as the total organic carbon concentration of the chemically solubilized lignite solution increased. Surprisingly, methane production using mMEC was drastically enhanced by approximately 50-fold when pulverized lignite was added as conductive material (CM) to the microcosms. To the best of our knowledge, this is the highest number of times methane production increased relative to the control. Our results demonstrated that bioaugmentation using a methanogenic consortium and adding pulverized lignite as CM could facilitate the bioconversion of chemically solubilized lignite solution to methane and lead to effective utilization of subterranean lignite, regarded as a neglected natural resource, without any further excavation processes.

Bacteria and Archaea Synergistically Convert Glycine Betaine to Biogenic Methane in the Formosa Cold Seep of the South China Sea.

Cold seeps are globally widespread seafloor ecosystems that feature abundant methane production and flourishing chemotrophic benthic communities. Chemical evidence indicates that cold seep methane is largely biogenic; however, the primary methane-producing organisms and associated pathways involved in methanogenesis remain elusive. This work detected methane production when glycine betaine (GBT) or trimethylamine (TMA) was added to the sediment microcosms of the Formosa cold seep, South China Sea. The methane production was suppressed by antibiotic inhibition of bacteria, while GBT was accumulated. This suggests that the widely used osmoprotectant GBT could be converted to cold seep biogenic methane via the synergistic activity of bacteria and methanogenic archaea because archaea are not sensitive to antibiotics and no bacteria are known to produce ample methane (mM). 16S rRNA gene diversity analyses revealed that the predominant bacterial and archaeal genera in the GBT-amended methanogenic microcosms included Oceanirhabdus and Methanococcoides. Moreover, metagenomic analyses detected the presence of grdH and mtgB genes that are involved in GBT reduction and demethylation, respectively. Two novel species were obtained, including bacterium Oceanirhabdus seepicola, which reduces GBT to TMA, and a methanogenic archaeon, Methanococcoides seepicolus, which produces methane from TMA and GBT. The two strains reconstituted coculture efficiently converted GBT to methane at 18°C; however, at 4°C addition of dimethylglycine (DMG), the GBT demethylation product, was necessary. Therefore, this work demonstrated that GBT is the precursor not only of the biogenic methane but also of the cryoprotectant DMG to the microorganisms at the Formosa cold seep. IMPORTANCE Numerous cold seeps have been found in global continental margins where methane is enriched in pore waters that are forced upward from sediments. Therefore, high concerns have been focused on the methane-producing organisms and the metabolic pathways in these environments because methane is a potent greenhouse gas. In this study, GBT was identified as the main precursor for methane in the Formosa cold seep of the South China Sea. Further, synergism of bacteria and methanogenic archaea was identified in GBT conversion to methane via the GBT reduction pathway, while methanogen-mediated GBT demethylation to methane was also observed. In addition, GBT-demethylated product dimethyl glycine acted as a cryoprotectant that promoted the cold seep microorganisms at cold temperatures. GBT is an osmoprotectant that is widely used by marine organisms, and therefore, the GBT-derived methanogenic pathway reported here could be widely distributed among global cold seep environments.

Alteration of Methanogenic Archaeon by Ethanol Contribute to the Enhancement of Biogenic Methane Production of Lignite.

Bioconverting coal to methane is a green and environmental friendly method to reuse waste coal. In this study, heterologous bacteria were used for the gas-producing fermentation of lignite under laboratory conditions, simultaneously, different concentrations of ethanol added into the culture to investigate the effect of ethanol on gas production and microbial flora structure. Results show that when the ethanol concentration was 1%, the best methanogenesis was achieved at 44.86 mL/g, which was twice the gas production of 0% ethanol. Before and after gas fermentation, the composition and structure of the coal changed, the volatile matter and fixed carbon increased, and the ash decreased. The absorbance value at characteristic peaks of all functional groups decreased, new peaks were generated at 2,300/cm, and the peak value disappeared at 3,375/cm. Thus, microorganisms interacted with coal, consumed it, and produced new materials. The microbial flora changes during gas production were tracked in real time. 0.5 and 1% ethanol did not obviously change the bacterial communities but strongly influenced the archaeon communities, thereby changed the methane production pathway. In the absence of ethanol, Methanosarcina was continuously increasing with the extension of fermentation time, this pathway was the nutrient type of acetic acid. When ethanol was added, Methanobacterium gradually increased, the pathway was mainly hydrotropic type. In summary, adding ethanol can increase the coalbed methane production, change the structure and composition of coal, and facilitate the interaction of microbe with coal. Therefore, the methanogenic archaeon changes could help improve the methane-producing ability of lignite in the presence of ethanol.

Microbial methanogenesis in subsurface oil and coal.

It is now clear that active methanogens are present in the deep-subsurface. This paper reviews microbial population structures and the biodegradation of organic compounds to methane in situ within oil reservoirs and coal deposits. It summarizes our current knowledge of methanogenes and methanogenesis, fermenters, synthrophs and microbial metabolism of complex organic compounds in these two widely occurring organic-rich subsurface environments. This review is not intended to be an exhaustive report of microbial diversity. Rather, it illustrates the similarities and differences between the two environments with specific examples, from the nature of the organic molecules to the methanogenic metabolic pathways and the structure of the microbial populations to demonstrate that widely diverging microbial populations show surprisingly similar metabolic capabilities.