The Variation of Microbial(Methanotroph)Communities in Marine Sediments Due to Aerobic Oxidation of Hydrocarbons

Methanotrophs in marine sediments and overlying water attenuate the emissions of methane into the atmosphere and thus play an important role for the global cycle of this greenhouse gas.However,gas released from natural hydrocarbon seeps are not pure methane but commonly mixed hydrocarbons.Currently,how methanotrophic bacteria behave in the co-presence of methane and heavier hydrocarbons remains unknown.In this paper,the bacteria were cultured aerobically in fresh sediment samples(collected from Bohai Bay in eastern China)at 28℃under the atmospheres of pure methane and methane+ethane+propane mixed gas,respec-tively.The prevailing terrigenous n-alkanes and fatty acids in the original sediment samples varied consistently after incubations,confirming the proceeding of aerobic bacterial activities.The real-time quantitative PCR assay and sequencing of the 16S rRNA and particulate methane monooxygenase(pmoA)genes revealed the changes of microbe communities to a methanotroph-dominating structure after incubations.Particularly,after incubations the family Methylococcaceae(typeⅠmethanotrophs)became dominant with proportions higher than 40%,whereas Methylocystaceae(typeⅡmethanotrophs)nearly disappeared in all incubated samples.More-over,the species of methanotrophs from the samples treated with pure methane were dominated by Methylobacter luteus,whereas Methylobacter whittenburyi took the predominant proportion in the samples treated with mixed gas.The phenomenon suggests that some methanotrophs may also utilize ethane and propane.Collectively,this study may help to gain a better understanding of the ef-fects and contributions of microbial activities in marine hydrocarbon seep ecosystems.

Evaluation Methods and Indexes for Ultra-deep Petroleum Reservoir Preservation with Case Studies

Following consideration of the characteristics of high temperature,high pressure and high in-situ stress in ultradeep sedimentary basins,together with the existence of hydrocarbon phase state transformation,hydrocarbon-water-rock interaction and rock mechanical property transition at those depths,the evaluation index system for hydrocarbon preservation was established.The physical leakage evaluation indexes can be divided into three categories:the dynamic efficiency indexes of micro-sealing,caprock integrity and natural gas diffusion.The chemical loss evaluation indexes can be divided into two categories:the thermochemical sulfate reduction(TSR)index in marine gypsum-bearing carbonate strata and the thermochemical oxidation of hydrocarbons(TOH)index in clastic strata.The slippage angle and overconsolidation ratio(OCR)are the key evaluation indexes in the evaluation of the integrity of shale caprocks.TSR intensity can be quantitatively calculated by use of the Zn PVT state parameter method.The TOH strength can be used to estimate the degree of hydrocarbon chemical loss,based on the TOH-related authigenic calcite cement content or the degree of negativeδ^(13)C of authigenic calcite.For the evaluation of ultra-deep preservation in specific areas,key indexes can be selected according to the local geological conditions,instead of all indexes needing to be evaluated for every scenario.