Distribution of the indigenous microorganisms and mechanisms of their orientational activation in Daqing Oilfield

The distribution of indigenous microorganisms was surveyed in Block 1 of Daqing Oilfield. Based on this survey, the indigenous microorganisms in the formation water were activated with different activator systems at the simulated stratum ecological environment. The changes of the number of bacteria of various physiological groups were determined during the process of activation. Also changes of pH value and composition of gas productions were analyzed at the end of culturing. The results showed that the selected block formation water contained a great number of saprophytic bacteria, hydrocarbon-oxidizing bacteria, fermentative bacteria, methane-producing bacteria and sulfate-reducing bacteria. Under the conditions that the growth of sulfate-reducing bacteria was controlled the block had the potential to enhance oil recovery by activating beneficial bacteria. The growth of sulfate-reducing bacteria can be inhibited through the activation of nitrate-reducing bacteria. The number of nitrate-reducing bacteria reached 10⁶–10⁷ cells/mL, but sulfate-reducing bacteria reached only 0–45 cells/mL in A system. Methane-producing bacteria can be activated by C, D activators. The relative content of biological methane in the light hydrocarbon gas reached 80% in C, D systems. B activator was conducive to the propagation of acid-producing bacteria, so that the pH value of the culture medium decreased from 7.5 to around 5.0. Hydrocarbon-oxidizing bacteria can be activated by various activator systems. There was low molecular light hydrocarbon in gas production according to the analysis of gas chromatograph. According to the content of methane and the number of methane-producing bacteria, methane only can be generated through activating methane-producing bacteria. By choosing different activator systems, various populations of indigenous microorganisms can be activated accordingly.

     

Microbiological Processes in the Gor'kovskoe Reservoir

The abundance of various groups of bacteria and the rate of microbiological processes of organic decay and methane cycle in the water and soils of the Gor'kovskoe Reservoir are determined, and the ecological characterization of its river reach is presented. The autochthonous and allochthonous impact on the bacterial community of the reservoir was found to be tolerable; however, some restricted areas were revealed, where the ecosystem is strongly affected by domestic and industrial wastewater, and where the abundance and activity of anaerobic methane-producing and sulfate-reducing soil bacteria increase.     

The origin mechanism of coalbed methane in the eastern edge of Ordos Basin

In the eastern edge of the Ordos Basin, the coalbed methane (CBM) development has not made substantial progress in the past 20 years, and the origin of gas can be used to guide the CBM block-selecting and development. Based on the 37 sets of carbon isotope data, the origin of the gas was determined and the origin mechanism was studied in this work. The δ ¹³C_PDB of methane ranges from ?70.5‰ to ?36.19‰ in the eastern edge in the Ordos Basin and the value becomes heavier from the north to the south. The secondary biogenic gas and the thermogenic gas are mixed in the shallow area and the thermogenic gas occurs in the medium and deep levels. The phenomenon is controlled mainly by the distribution of coal rank and hydrodynamics. Firstly, based on the relationship between China coal rank and methane δ ¹³C_PDB, the medium rank coal is dominant in the eastern edge of the Ordos Basin, and the mixture of the secondary biogenic gas and the thermogenic gas is formed in the coal of vitrinite reflectant ratio (R _max) between 0.5% and 2.0% if there is appropriate hydrodynamics; at the same time, because of the shallow burial depth, and the well-developed coal outcrop, meteoric water and other surface water carrying bacteria recharge the coal reservoir, metabolize the organic compounds at a relatively low temperature, and generate methane and carbon dioxide. Wherever the trapping mechanisms occur in the coal, such as Liulin and Hancheng, modern gas content should be high.     

Clayey materials in river basin enhancing microbial contamination of river water

Mineral constituents of clay materials may promote interaction, adsorption and attachment of microorganisms, often resulting in biofilms’ formation. In this study investigation is made to determine how littoral clayey materials on the shores of a river promote accumulation of bacteria and increase contamination of river water. Clayey samples were collected at various points along the shore of a river around Mondeor in Johannesburg and the mineralogical composition was determined using XRD and XRF. Microorganisms in clay-biofilm and river water were identified by DNA sequencing and plate count. Results showed that total coliforms, Escherichia coli, Pseudomonas sp. and presumptive indigenous microorganisms attached to littoral clayey materials containing the mineral muscovite (characterising argillaceous soils). Bacteria number on clayey materials was significantly higher than on overlying water especially before rainy season. However a decrease of the number of bacteria in clayey materials concurrent with an increase in the number of suspended bacteria after rain events, was the result of the action of high and fast flows in the basin, eroding the biofilms. Attachment of microorganisms in clayey material as observed in this study could be ascribed to the glue-like aspect of soil (due to muscovite) that facilitates adhesion. It therefore demonstrates the potential of clayey materials to encourage biofilm formation and enhance microbial contamination of river water as shown here.     

The Effect of Fatty Acid Diffusion in Leachate on the Propagation of Concentration Waves in the Process of Municipal Solid Waste Decomposition

A distributed model of municipal solid waste decomposition is proposed. The main model parameters are concentrations of municipal solid waste, volatile fatty acids, and the biomass of methane-producing microorganisms. Numerical analysis of the model is made. Diffusion of fatty acids in leachate, which facilitates the formation of anaerobic conditions, is shown to cause the formation of concentration chemical waves propagating in the space. The area of initiation methane production is found to expand.     

城市河流沉积物中氮素与好氧甲烷氧化菌群落特征响应关系

由于人类活动的影响大量未经处理的废污水汇入城市河流,高浓度的污染物影响了河流中微生物对生物地球化学物质迁移和转化的介导作用.本文选取典型的城市河流——北运河作为研究区域,分析了北运河沉积物中氮素形态以及含量的空间和季节差异性,并结合克隆文库分子生物学的方法,探讨了氮素形态和含量的差异对好氧甲烷氧化菌(aerobic methane-oxidizing microorganisms,MOB)群落特征的影响.结果表明:北运河沉积物中铵态氮(NH_4~+-N)为氮素的主要存在形态,存在显著的空间差异,其含量在下游显著高于上游,但季节差异不显著.NH_4~+-N含量的空间差异对MOB的群落结构和群落分布有显著影响,对群落多样性影响不显著.NH_4~+-N含量的空间分布特征与MOB的群落聚类特征一致,NH_4~+-N对MOB群落分布的影响显著高于其他形态的氮素,其含量越高,则与MOB群落分布的响应关系越紧密.北运河中NH_4~+-N的来源影响了沉积物中MOB的主要来源,MOB高同源性菌群的来源与NH_4~+-N等主要污染物的来源一致.沉积物中MOB物种之间联系的紧密程度依赖于氮素的主要存在形态及其含量水平.NH_4~+-N含量较高的下游沉积物中微生物彼此之间关系及集聚程度更强,受外界环境变化的干扰程度更强,受人类活动引起环境变化的敏感程度更高.城市河流中氮素的形态和含量差异对甲烷的氧化过程有显著影响.探究城市河流沉积物中高含量的NH_4~+-N对甲烷产生及消耗的影响过程是控制河流温室气体排放的关键.     

Inspirations from the scientific discovery of the anammox bacteria: A classic example of how scientific principles can guide discovery and development

Anaerobic ammonium oxidation(anammox) is a relatively new pathway within the N cycle discovered in the late 1990 s. This eminent discovery not only modified the classical theory of biological metabolism and matter cycling, but also profoundly influenced our understanding of the energy sources for life. A new member of chemolithoautotrophic microorganisms capable of carbon fixation was found in the vast deep dark ocean. If the discovery of the chemosynthetic ecosystems in the deep-sea hydrothermal vent environments once challenged the old dogma "all living things depend on the sun for growth," the discovery of anammox bacteria that are widespread in anoxic environments fortifies the victory over this dogma. Anammox bacteria catalyze the oxidization of NH_4~+ by using NO_2~- as the terminal electron acceptor to produce N_2. Similar to the denitrifying microorganisms, anammox bacteria play a biogeochemical role of inorganic N removal from the environment. However, unlike heterotrophic denitrifying bacteria, anammox bacteria are chemolithoautotrophs that can generate transmembrane proton motive force, synthesize ATP molecules and further carry out CO_2 fixation through metabolic energy harvested from the anammox process. Although anammox bacteria and the subsequently found ammonia-oxidizing archaea(AOA), another very important group of N cycling microorganisms are both chemolithoautotrophs, AOA use ammonia rather than ammonium as the electron donor and O_2 as the terminal electron acceptor in their energy metabolism. Therefore, the ecological process of AOA mainly takes place in oxic seawater and sediments, while anammox bacteria are widely distributed in anoxic water and sediments, and even in some typical extreme marine environments such as the deep-sea hydrothermal vents and methane seeps. Studies have shown that the anammox process may be responsible for 30%–70% N_2 production in the ocean. In environmental engineering related to nitrogenous wastewater treatment, anammox provides a new technology with low energy consumption, low cost, and high efficiency that can achieve energy saving and emission reduction. However, the discovery of anammox bacteria is actually a hard-won achievement. Early in the 1960 s, the possibility of the anammox biogeochemical process was predicted to exist according to some marine geochemical data. Then in the 1970 s, the existence of anammox bacteria was further predicted via chemical reaction thermodynamic calculations. However, these microorganisms were not found in subsequent decades. What hindered the discovery of anammox bacteria, an important N cycling microbial group widespread in hypoxic and anoxic environments? What are the factors that finally led to their discovery? What are the inspirations that the analyses of these questions can bring to scientific research? This review article will analyze and elucidate the above questions by presenting the fundamental physiological and ecological characteristics of the marine anammox bacteria and the principles of scientific research.     

Hydrocarbon status in Florida real estate canals

The level of hydrocarbons present in the sediments of real estate canals on the east and west coast of Florida was determined using gas chromatography. Although the amount of alkanes present was found to be comparatively low, the ratio of the amount of odd carbon-numbered alkanes to even carbon-numbered alkanes suggested that canals on the Gulf of Mexico are receiving an influx of petroleum products. In addition, the most probable number technique was used to enumerate total aerobic heterotrophic bacteria, aerobic hydrocarbon-degrading bacteria, sulfate reducing bacteria, and sulfate-reducing hydrocarbon-degrading bacteria present in the canal sediments. Results showed significantly higher numbers of hydrocarbon degrading bacteria in the sediments of the west coast canals.     

Quantification of seep-related methane gas emissions at Tommeliten,North Sea

Tommeliten is a prominent methane seep area in the Central North Sea. Previous surveys revealed shallow gas-bearing sediments and methane gas ebullition into the water column. In this study, the in situ methane flux at Tommeliten is re-assessed and the potential methane transport to the atmosphere is discussed, with regards to the hydrographic setting and gas bubble modeling. We have compiled previous data, acquired new video and acoustic evidence of gas bubble release, and have measured the methane concentration, and its C-isotopic composition in the water column. Parametric subbottom sonar data reveal the three-dimensional extent of shallow gas and morphologic features relevant for gas migration. Five methane ebullition areas are identified and the main seepage area appears to be 21 times larger than previously estimated. Our video, hydroacoustic, subbottom, and chemical data suggest that ～1.5×10⁶ mol CH₄/yr (～26 tons CH₄/yr) of methane gas is being released from the seepage area of Tommeliten. Methane concentration profiles in the vicinity of the gas seeps show values of up to 268 nM (～100 times background) close to the seafloor. A decrease in δ¹³C-CH₄ values at 40 m water depth indicates an unknown additional biogenic methane source within the well oxygenated thermocline between 30 and 40 m water depth. Numerical modeling of the methane bubbles due to their migration and dissolution was performed to estimate the bubble-derived vertical methane transport, the fate of this methane in the water column, and finally the flux to the atmosphere. Modeling indicates that less than ～4% of the gas initially released at the seafloor is transported via bubbles into the mixed layer and, ultimately, to the atmosphere. However, because of the strong seasonality of mixing in the North Sea, this flux is expected to increase as mixing increases, and almost all of the methane released at the seafloor could be transferred into the atmosphere in the stormy fall and winter time.     

Greenhouse gas emissions (CO2, CH4, and N2O) from several perialpine and alpine hydropower reservoirs by diffusion and loss in turbines

We investigated greenhouse gas emissions (CO₂, CH₄, and N₂O) from reservoirs located across an altitude gradient in Switzerland. These are the first results of greenhouse gas emissions from reservoirs at high elevations in the Alps. Depth profiles were taken in 11 reservoirs located at different altitudes between the years 2003 and 2006. Diffusive trace gas emissions were calculated using surface gas concentrations, wind speeds and transfer velocities. Additionally, methane entering with the inflowing water and methane loss at the turbine was assessed for a subset of the reservoirs. All reservoirs were emitters of carbon dioxide and methane with an average of 970?±?340?mg?m^?2?day^?1 (results only from four lowland and one subalpine reservoir) and 0.20?±?0.15?mg?m^?2?day^?1, respectively. One reservoir (Lake Wohlen) emitted methane at a much higher rate (1.8?±?0.9?mg?m^?2?day^?1) than the other investigated reservoirs. There was no significant difference in methane emissions across the altitude gradient, but average dissolved methane concentrations decreased with increasing elevation. Only lowland reservoirs were sources for N₂O (72?±?22???g?m^?2?day^?1), while the subalpine and alpine reservoirs were in equilibrium with atmospheric concentrations. These results indicate reservoirs from subalpine/alpine regions to be only minor contributors of greenhouse gases to the atmosphere compared to other reservoirs.     

Evidence of methylmercury production and modification of the microbial community structure in estuary sediments contaminated with wastewater treatment plant effluents

The Seine’s estuary (France) waters are the receptacle of effluents originating from wastewater treatment plants (WWTP). In this estuary, mudflats are deposition zones for sediments and their associated contaminants, and play an essential role in the mercury (Hg) biogeochemical cycle mainly due to indigenous microorganisms. Microcosms were used to assess the impact of WWTP-effluents on mercury methylation by monitoring Hg species (total dissolved Hg in porewater, methylmercury and total mercury) and on microbial communities in sediments. After effluent amendment, methylmercury (MeHg) concentrations increased in relation with the total Hg and organic matter content of the WWTP-effluents. A correlation was observed between MeHg and acid-volatile-sulfides concentrations. Quantification of sulfate-reducing microorganisms involved in Hg methylation showed no increase of their abundance but their activity was probably enhanced by the organic matter supplied with the effluents. WWTP-effluent spiking modified the bacterial community fingerprint, mainly influenced by Hg contamination and the organic matter amendment.     

内陆水体好氧甲烷氧化过程研究进展

内陆水体是全球碳循环的关键组成部分,是大气中甲烷(CH4)的重要来源,每年从内陆淡水与自然湿地排放进入大气的CH4约为185～357 Tg/a.通常,内陆水体中CH4主要由分布于水层底部的厌氧区或沉积层内的产甲烷菌介导产生,其向水层表面传输的过程中易被甲烷氧化菌所氧化.甲烷氧化菌可分为好氧甲烷氧化菌和厌氧甲烷氧化菌,有...     

Assessing Methane in Shallow Groundwater in Unconventional Oil and Gas Play Areas,Eastern Kentucky

The expanding use of horizontal drilling and hydraulic fracturing technology to produce oil and gas from tight rock formations has increased public concern about potential impacts on the environment, especially on shallow drinking water aquifers. In eastern Kentucky, horizontal drilling and hydraulic fracturing have been used to develop the Berea Sandstone and the Rogersville Shale. To assess baseline groundwater chemistry and evaluate methane detected in groundwater overlying the Berea and Rogersville plays, we sampled 51 water wells and analyzed the samples for concentrations of major cations and anions, metals, dissolved methane, and other light hydrocarbon gases. In addition, the stable carbon and hydrogen isotopic composition of methane (δ¹³C‐CH₄ and δ²H‐CH₄) was analyzed for samples with methane concentration exceeding 1 mg/L. Our study indicates that methane is a relatively common constituent in shallow groundwater in eastern Kentucky, where methane was detected in 78% of the sampled wells (40 of 51 wells) with 51% of wells (26 of 51 wells) exhibiting methane concentrations above 1 mg/L. The δ¹³C‐CH₄ and δ²H‐CH₄ ranged from ?84.0‰ to ?58.3‰ and from ?246.5‰ to ?146.0‰, respectively. Isotopic analysis indicated that dissolved methane was primarily microbial in origin formed through CO₂ reduction pathway. Results from this study provide a first assessment of methane in the shallow aquifers in the Berea and Rogersville play areas and can be used as a reference to evaluate potential impacts of future horizontal drilling and hydraulic fracturing activities on groundwater quality in the region.     

Higher effectiveness of photoinactivation of bacterial spores,UV resistant vegetative bacteria and mold spores with 222 nm compared to 254 nm wavelength

Eleven selected species of vegetative bacteria, bacteria spores and mold spores were irradiated with different doses of UV radiation of a 222 nm krypton‐chloride excimer lamp and a 254 nm mercury lamp under laboratory conditions. Then the inactivation curves were determined. The necessary UV fluences for a respective reduction were higher for the excimer lamp for the tested vegetative bacteria of Bacillus cereus, Arthrobacter nicotinovorans, Staphylococcus aureus and Pseudomonas aeruginosa and slightly higher for the spores of Streptomyces griseus and Clostridium pasteurianum. However, less than 250 J/m² UV fluence with 222 nm was sufficient for a 4‐log reduction, depending on the species. On the other hand, the UV fluences for the 254 nm mercury lamp were much higher for the bacterial spores of Bacillus cereus, Thermoactinomyces griseus and the bacteria of Deinococcus radiodurans and slightly higher for the mold spores of Aspergillus niger and Penicillium expansum. The results show that especially the germs with a higher UV resistance and those with more effective repair mechanisms can be inactivated more efficiently by the 222 nm excimer lamp. This may be due to the fact that low UV fluence mainly affects radiation sensitive microorganisms by DNA damage whereas at higher UV fluence (various) mechanisms of protein damage can presumably be held responsible for inactivation.     

Using DNA‐Stable Isotope Probing to Identify MTBE‐ and TBA‐Degrading Microorganisms in Contaminated Groundwater

Although the anaerobic biodegradation of methyl tert‐butyl ether (MTBE) and tert‐butyl alcohol (TBA) has been documented in the laboratory and the field, knowledge of the microorganisms and mechanisms involved is still lacking. In this study, DNA‐stable isotope probing (SIP) was used to identify microorganisms involved in anaerobic fuel oxygenate biodegradation in a sulfate‐reducing MTBE and TBA plume. Microorganisms were collected in the field using Bio‐Sep® beads amended with ¹³C₅‐MTBE, ¹³C₁‐MTBE (only methoxy carbon labeled), or ¹³C₄‐TBA. ¹³C‐DNA and ¹²C‐DNA extracted from the Bio‐Sep beads were cloned and 16S rRNA gene sequences were used to identify the indigenous microorganisms involved in degrading the methoxy group of MTBE and the tert‐butyl group of MTBE and TBA. Results indicated that microorganisms were actively degrading ¹³C‐labeled MTBE and TBA in situ and the ¹³C was incorporated into their DNA. Several sequences related to known MTBE‐ and TBA‐degraders in the Burkholderiales and the Sphingomonadales orders were detected in all three ¹³C clone libraries and were likely to be primary degraders at the site. Sequences related to sulfate‐reducing bacteria and iron‐reducers, such as Geobacter and Geothrix, were only detected in the clone libraries where MTBE and TBA were fully labeled with ¹³C, suggesting that they were involved in processing carbon from the tert‐butyl group. Sequences similar to the Pseudomonas genus predominated in the clone library where only the methoxy carbon of MTBE was labeled with ¹³C. It is likely that members of this genus were secondary degraders cross‐feeding on ¹³C‐labeled metabolites such as acetate.     

Hydrocarbon‐Rich Groundwater above Shale‐Gas Formations: A Karoo Basin Case Study

Horizontal drilling and hydraulic fracturing have enhanced unconventional hydrocarbon recovery but raised environmental concerns related to water quality. Because most basins targeted for shale‐gas development in the USA have histories of both active and legacy petroleum extraction, confusion about the hydrogeological context of naturally occurring methane in shallow aquifers overlying shales remains. The Karoo Basin, located in South Africa, provides a near‐pristine setting to evaluate these processes, without a history of conventional or unconventional energy extraction. We conducted a comprehensive pre‐industrial evaluation of water quality and gas geochemistry in 22 groundwater samples across the Karoo Basin, including dissolved ions, water isotopes, hydrocarbon molecular and isotopic composition, and noble gases. Methane‐rich samples were associated with high‐salinity, NaCl‐type groundwater and elevated levels of ethane, ⁴He, and other noble gases produced by radioactive decay. This endmember displayed less negative δ¹³C‐CH₄ and evidence of mixing between thermogenic natural gases and hydrogenotrophic methane. Atmospheric noble gases in the methane‐rich samples record a history of fractionation during gas‐phase migration from source rocks to shallow aquifers. Conversely, methane‐poor samples have a paucity of ethane and ⁴He, near saturation levels of atmospheric noble gases, and more negative δ¹³C‐CH₄; methane in these samples is biogenic and produced by a mixture of hydrogenotrophic and acetoclastic sources. These geochemical observations are consistent with other basins targeted for unconventional energy extraction in the USA and contribute to a growing data base of naturally occurring methane in shallow aquifers globally, which provide a framework for evaluating environmental concerns related to unconventional energy development (e.g., stray gas).     

人工湿地植物根区微生物与净化效果的季节变化

人工湿地污水处理系统具有净化效果显著、建设和运行费用低廉、管理简便等优点.近年来越来越受到人们的重视.人工湿地生态系统受环境影响较大,研究人工湿地在不同季节的净化功能对于充分发挥其作用具有重要意义.对不同季节的芦苇(Phragmite saus-tralis)、茭白(Zizania caduciflora)和香蒲(Typha latifolia)等湿地系统植物根区、无植物对照基质微生物分布状况以及净化效果进行了研究,结果显示:(1)种植不同植物的湿地系统根区微生物数量不同,其湿地净化效果也不同;(2)季节变化显示湿地系统秋季的细菌总数量比夏季高;但对照系统夏季的真菌和放线菌数量明显高于秋季;(3)湿地系统总磷(TP)的去除率夏季高于秋季;(4)相关性分析发现湿地植物根区的细菌总数与BOD5的去除率之间存在显著相关性;而根区微生物数量与TP、CODCr和凯氏氮(KN)的去除率之间不存在显著相关性,具体机理还需进一步研究.     

Simulation experiments on gas production from hydrate-bearing sediments

Experiments were made on 58 sediment samples from four sites(1244,1245,1250 and 1251) of ODP204 at five temperature points(25,35,45,55 and 65℃) to simulate methane production from hydrate-bearing sediments.Simulation results from site 1244 show that the gas components consist mainly of methane and carbon dioxide,and heavy hydrocarbons more than C2+ cannot be detected.This site also gives results,similar to those from the other three,that the methane production is controlled by experimental temperatures,gene...     

Factors controlling the noble gas abundance patterns of deep-sea basalts

A number of processes may modify the noble gas composition of silicate liquids so that the composition of noble gases observed in glassy margins of deep-sea basalts is not that of the upper mantle. Differential solubility enhances the light noble gases relative to the heavier gases; however, we demonstrate that the observed abundance pattern cannot be attributed to solubility of noble gases with atmospheric proportions. Partial melting and fractional crystallization increase the noble gas content of all species relative to mantle concentrations, but do not fractionate their relative abundances. Noble gases may be lost from an ascending magma in various ways, the most important, however, may be exclusion of gas from crystals forming at the time of solidification, which is shown to result in marked loss of gas from the basalt. Small amounts of low-temperature alteration of solidified basalt can produce dramatic changes in the noble gas abundance pattern, since the adsorption coefficients for the different noble gas favor uptake of heavy species relative to the light species. Atmospheric contamination can account for observed variations in the ⁴⁰Ar/³⁶Ar ratio of oceanic basalts. The degree of crystallinity of glassy margins of deep-sea basalts may control the helium abundance of these samples; however, the uniform ³He/⁴He values reported apparently reflect a relatively constant proportion of radiogenic and primordial helium in the mantle.     

Light carbon isotope events of foraminifera attributed to methane release from gas hydrates on the continental slope,northeastern South China Sea

In 2013, the China Geological Survey and Guangzhou Marine Geological Survey conducted the second Chinese gas hydrate expedition in the northern South China Sea(SCS) and successfully obtained visible gas hydrate samples. Five of the thirteen drilling sites were cored for further research. In this work, Site GMGS2-08 is selected for the stable isotopic analysis of foraminifera present in the boreholes in order to reveal the carbon isotopic characteristics of the foraminifera and their response to methane release in the gas hydrate geological system. Our results show that the methane content at Site GMGS2-08 is extremely high, with headspace methane concentrations up to 39300 μmol L~(-1). The hydrocarbon δ~(13)C values, ranging from-69.4‰ to-72.3‰ PDB, distinctly indicate biogenic generation. Based on the δD analytical results(~(-1)83‰ to~(-1)85‰ SMOW), headspace methane is further discriminated to be microbial gas, derived from CO_2 reduction. By isotopic measurement, five light δ~(13)C events are found in the boreholes from Site GMGS2-08, with foraminiferal δ~(13)C values being apparently lower than the normal variation range found in the glacial-interglacial cycles of the SCS. The δ~(13)C values of benthic Uvigerina peregrina are extremely depleted(as low as~(-1)5.85‰ PDB), while those of planktonic Globigerinoides ruber reach-5.68‰ PDB. Scanning electron micrograph(SEM) studies show that foraminiferal tests have experienced post-depositional alteration, infilled with authigenic carbonate, and the diagenetic mineralization is unlikely to be related to the burial depths. The correlation calculation suggests that the anaerobic oxidation of organic matter has only weak influences on the δ~(13)C composition of benthic foraminifera. This means that the anomalous δ~(13)C depletions are predominantly attributed to the overprinting of secondary carbonates derived from the anaerobic oxidation of methane(AOM). Furthermore, the negative δ~(13)C anomalies, coupled with the positive δ18O anomalies observed at Site GMGS2-08, are most likely the critical pieces of evidence for gas hydrate dissociation in the geological history of the study area.