石油烃的厌氧生物降解对油藏残余油气化开采的启示

利用微生物将油藏中难以动用的原油就地转化为甲烷,以天然气的形式开采、或作为战略资源就地储备,从而大幅度提高油气资源的利用率,是当前国际上研究的前沿课题。本文综述了石油烃厌氧生物降解转化为甲烷的菌群结构、反应热力学和反应动力学等基础科学问题的最新研究进展,讨论了油藏残余油气化开采技术的可行性及开发潜力,提出了该技术进一步研究的方向。     

Methanogens and sulfate-reducing bacteria in oil sands fine tailings waste

In the past decade, the large tailings pond (Mildred Lake Settling Basin) on the Syncrude Canada Ltd. lease near Fort McMurray, Alta., has gone methanogenic. Currently, about 60%-80% of the flux of gas across the surface of the tailings pond is methane. As well as adding to greenhouse gas emissions, the production of methane in the fine tailings zone of this and other settling basins may affect the performance of these settling basins and impact reclamation options. Enumeration studies found methanogens (10(5)-10(6) MPN/g) within the fine tailings zone of various oil sands waste settling basins. SRB were also present (10(4)-10(5) MPN/g) with elevated numbers when sulfate was available. The methanogenic population was robust, and sample storage up to 9 months at 4 degrees C did not cause the MPN values to change. Nor was the ability of the consortium to produce methane delayed or less efficient after storage. Under laboratory conditions, fine tailings samples released 0.10-0.25 mL CH4 (at STP)/mL fine tailings. The addition of sulfate inhibited methanogenesis by stimulating bacterial competition.     

Metagenomic analysis for the microbial consortium of anaerobic CO oxidizers

Metagenomics analysis has been applied to identify the dominant anaerobic microbial consortium of the carbon monoxide (CO) oxidizers in anaerobic sludge. Reads from the hypervariable V6 region in the bacterial 16s rDNA were aligned and finally clustered into operational taxonomic units (OTUs). The OTUs from different stages in anaerobic CO condition were classified. Alphaproteobacteria, clostridia, betaproteobacteria and actinobacteria were the most abundant groups, while alphaproteobacteria, betaproteobacteria and actinobacteria were variable groups. CO consumption and production efficiency of the microbial consortium were studied. Semi-continuous trials showed that these anaerobic CO oxidizers formed a stable microbial community, and the CO conversion rate was at over 84%, with the highest CO consumption activity of 28.9 mmol CO/g VSS●day and methane production activity at 7.6 mmol CH₄/g VSS●day during six cycles.     

Activity, distribution, and abundance of methane-oxidizing bacteria in the near surface soils of onshore oil and gas fields

Methane-oxidizing bacteria (MOB) have long been used as an important biological indicator for oil and gas prospecting, but the ecological characteristics of MOB in hydrocarbon microseep systems are still poorly understood. In this study, the activity, distribution, and abundance of aerobic methanotrophic communities in the surface soils underlying an oil and gas field were investigated using biogeochemical and molecular ecological techniques. Measurements of potential methane oxidation rates and pmoA gene copy numbers showed that soils inside an oil and gas field are hot spots of methane oxidation and MOB abundance. Correspondingly, terminal restriction fragment length polymorphism analyses in combination with cloning and sequencing of pmoA genes also revealed considerable differences in the methanotrophic community composition between oil and gas fields and the surrounding soils. Principal component analysis ordination furthermore indicated a coincidence between elevated CH₄ oxidation activity and the methanotrophic community structure with type I methanotrophic Methylococcus and Methylobacter, in particular, as indicator species of oil and gas fields. Collectively, our results show that trace methane migrated from oil and gas reservoirs can considerably influence not only the quantity but also the structure of the methanotrophic community.     

Nitrogen removal from purified swine wastewater using biogas by semi-partitioned reactor

Nitrate and ammonium removal from purified swine wastewater using biogas and air was investigated in continuous reactor operation. A novel type of reactor, a semi-partitioned reactor (SPR), which enables a biological reaction using methane and oxygen in the water phase and discharges these unused gases separately, was operated with a varying gas supply rate. Successful removal of NO(3)(-) and NH(4)(+) was observed when biogas and air of 1L/min was supplied to an SPR of 9L water phase with a NO(2,3)(-)-N and NH(4)(+)-N removal rate of 0.10 g/L/day and 0.060 g/L/day, respectively. The original biogas contained an average of 77.2% methane, and the discharged biogas from the SPR contained an average of 76.9% of unused methane that was useable for energy like heat or electricity production. Methane was contained in the discharged air from the SPR at an average of 2.1%. When gas supply rates were raised to 2L/min and the nitrogen load was increased, NO(3)(-) concentration was decreased, but NO(2)(-) accumulated in the reactor and the NO(2,3)(-)-N and NH(4)(+)-N removal activity declined. To recover the activity, lowering of the nitrogen load and the gas supply rate was needed. This study shows that the SPR enables nitrogen removal from purified swine wastewater using biogas under limited gas supply condition.     

Energy Return on Investment (EROI) for Forty Global Oilfields Using a Detailed Engineering-Based Model of Oil Production

Studies of the energy return on investment (EROI) for oil production generally rely on aggregated statistics for large regions or countries. In order to better understand the drivers of the energy productivity of oil production, we use a novel approach that applies a detailed field-level engineering model of oil and gas production to estimate energy requirements of drilling, producing, processing, and transporting crude oil. We examine 40 global oilfields, utilizing detailed data for each field from hundreds of technical and scientific data sources. Resulting net energy return (NER) ratios for studied oil fields range from ≈2 to ≈100 MJ crude oil produced per MJ of total fuels consumed. External energy return (EER) ratios, which compare energy produced to energy consumed from external sources, exceed 1000:1 for fields that are largely self-sufficient. The lowest energy returns are found to come from thermally-enhanced oil recovery technologies. Results are generally insensitive to reasonable ranges of assumptions explored in sensitivity analysis. Fields with very large associated gas production are sensitive to assumptions about surface fluids processing due to the shifts in energy consumed under different gas treatment configurations. This model does not currently include energy invested in building oilfield capital equipment (e.g., drilling rigs), nor does it include other indirect energy uses such as labor or services.     

Bioconversion of natural gas to liquid fuel: Opportunities and challenges

Natural gas is a mixture of low molecular weight hydrocarbon gases that can be generated from either fossil or anthropogenic resources. Although natural gas is used as a transportation fuel, constraints in storage, relatively low energy content (MJ/L), and delivery have limited widespread adoption. Advanced utilization of natural gas has been explored for biofuel production by microorganisms. In recent years, the aerobic bioconversion of natural gas (or primarily the methane content of natural gas) into liquid fuels (Bio-GTL) by biocatalysts (methanotrophs) has gained increasing attention as a promising alternative for drop-in biofuel production. Methanotrophic bacteria are capable of converting methane into microbial lipids, which can in turn be converted into renewable diesel via a hydrotreating process. In this paper, biodiversity, catalytic properties and key enzymes and pathways of these microbes are summarized. Bioprocess technologies are discussed based upon existing literature, including cultivation conditions, fermentation modes, bioreactor design, and lipid extraction and upgrading. This review also outlines the potential of Bio-GTL using methane as an alternative carbon source as well as the major challenges and future research needs of microbial lipid accumulation derived from methane, key performance index, and techno-economic analysis. An analysis of raw material costs suggests that methane-derived diesel fuel has the potential to be competitive with petroleum-derived diesel.     

Performance of anaerobic baffled reactor (ABR) treating synthetic wastewater containing p-nitrophenol

In this study, the effect of increasing p-nitrophenol (PNP) concentrations on the performance of anaerobic baffled reactor (ABR) (chemical oxygen demand (COD), removals, volatile fatty acid (VFA), p-aminophenol (PAP) and methane gas productions) was investigated through 240 days. The PNP concentrations were raised to 700 from 10 mg/L corresponding to PNP loading rates of 0.97 and 67.9 g/m³ day. The PNP and COD removal efficiencies were 99 and 90% at PNP loading rates as high as 33.9 g/m³ day, respectively, through the acclimation of anaerobic granular sludge. After this loading rate, the removal efficiencies decreased to 79%. The COD removal efficiencies were high in compartment 1 (E = 78–93%) while a small amount of COD removal was achieved in compartments 2 and 3. The PNP removal efficiencies were approximately 90% in all PNP loading rates except for loading rate of 0.97 g/m³ day. The maximum PNP removal efficiency was measured as 99% at a loading rate of 8.32 g/m³ day. The optimum PNP loading rate for maximum COD, PNP removals and methane yield was 8.32 g/m³ day. The total, methane gas productions and methane percentages were approximately 2160–2400 mL/day and 950–1250 mL/day and 44–52% for the PNP loading rates varying between 4.36 and 33.9 g/m³ day, respectively. For PNP loading rates varying between 33.9 and 67.9 g/m³ day, the total, methane gas productions and methane percentages were approximately 2160 and 960 mL/day and 44%, respectively. The highest total volatile fatty acid (TVFA) concentrations were found in the first compartment with fluctuated values varied between 50 and 200 mg/L indicating the acidogenesis. p-Aminophenol was found as the main intermediate through anaerobic degradation of PNP which later was broken down to phenol and ammonia.     

Methane fermentation of brewery by-products

A laboratory-scale research program was undertaken to investigate the kinetics of the mesophilic (37°C) anaerobic digestion of brewery industry by-product. The purpose was to develop data for the design and operation of full-scale units which could be used to generate methane fuel gas from these materials. This is important because the brewery industry has been susceptible to shortages of natural gas in recent years. The minimum SRT is 2.3 days, although for design purposes as much as ten days is recommended. The biomass yield is 0.512 g volatile suspended solids (VSS)/g volatile solids (VS) or 0.421 g VSS/g chemical oxygen demand (COD). The maintenance requirement is 0.052 g VS/g VSS per day or 0.061 g COD/g VSS per day. The specific methane yield is 2.51 liter/g VSS, and the methane productivity is 0.32–0.41 liter/g dry substrate added or 0.69–0.91 liter/g destroyed. The maximum loading rate for which substrate inhibition is not observed is 6 g dry substrate added per liter per day. The results of the entire program indicate that processing brewery by-product in this manner is both technically feasible and economically attractive.     

Performance characteristics of a two-stage dark fermentative system producing hydrogen and methane continuously

The performance of a mesophilic two-stage system generating hydrogen and methane continuously from sucrose (10-30 g/L) was investigated. A hydrogen-generating CSTR followed by an upflow anaerobic filter were both inoculated with anaerobically digested sewage sludge, and ORP, pH, gas output, %H(2), %CH(4) and %CO(2) monitored. pH was controlled with NaOH, KOH or Ca(OH)(2). Using NaOH as alkali with 10 g/L sucrose, yields of 1.62 +/- 0.2 mol H(2)/mol hexose added and 323 mL CH(4)/gCOD added to the hydrogen and methane reactors respectively were achieved. The overall chemical oxygen demand (COD) reduction was 92.6% with 0.90 +/- 0.1 g/L sodium and 316 +/- 40 mg/L residual acetate in the methane reactor. Operation at 20 g/L sucrose and NaOH as alkali led to impaired volatile fatty acid (VFA) degradation in the methane reactor with 2.23 +/- 0.2 g/L sodium, 1,885 mg/L residual acetate, a hydrogen yield of 1.47 +/- 0.1 mol/mol hexose added, a methane yield of 294 mL/gCOD added and an overall COD reduction of 83%. Using Ca(OH)(2) as alkali with 20 g/L sucrose gave a hydrogen yield of 1.29 +/- 0.3 mol/mol hexose added, a methane yield of 337 mL/gCOD added and improved the overall COD reduction to 91% with residual acetate concentrations of 522 +/- 87 mg/L. Operation at 30 g/L sucrose with Ca(OH)(2) gave poorer overall COD reduction (68%), a hydrogen yield of 1.47 +/- 0.2 mol/mol hexose added, a methane yield of 138 mL/gCOD added and residual acetate 7,343 +/- 715 mg/L. It was shown that sodium toxicity and overloading are important issues for successful anaerobic digestion of effluent from biohydrogen reactors in high rate systems.     

Anaerobic degradation of sulphite evaporator condensate in a fixed-bed loop reactor by a defined bacterial consortium

Summary After elucidating the composition of an anaerobic bacterial enrichment culture treating sulphite evaporator condensate (SEC), an effluent in the pulp and paper industry, we built up stepwise a defined mixed culture to convert the organic constituents of SEC (acetate, methanol, furfural) to methane and CO₂. In batch cultures Desulfovibrio furfuralis and Methanobacterium bryantii degraded furfural in the absence of sulphate via inter-species H₂ transfer yielding 0.42 mol methane and 1.87 mol acetate/mol furfural degraded. When Methanosarcina barkeri was added to this diculture, acetate was also transformed to methane yielding 0.93 mol methane/mol acetate converted. This consortium (D. furfuralis, Methanobacterium bryantii and Methanosarcina barkeri) degraded furfural in continuous culture (fixed-bed loop reactor) to 92%, but the conversion of acetate was only 67%. The conversion of acetate could be further improved to 86% by adding 10 mm sulphate to the medium. This resulted in a space time yield of 10.9 g chemical oxygen demand (COD)/1 per day for the overall conversion. With a consortium consisting of M. barkeri, Methanobrevibacter arboriphilus, Methanosaeta concilii and D. furfuralis, a synthetic SEC could be degraded at a space time yield of 13.35 g COD/1 per day. This defined culture degraded all the constituents of SEC at an efficiency of almost 90% compared to an enrichment culture under identical conditions.Offprint requests to: U. Ney     

Metabolism of chlorinated methanes,ethanes, and ethylenes by a mixed bacterial culture growing on methane

Summary Soil was taken from the top 10 cm of a soil column that removed halogenated aliphatic hydrocarbons in the presence of natural gas. This soil was used as an enrichment inoculum to determine that the removals seen in the soil column were in fact of a microbiological nature. Methane served as the source of carbon and energy and was consumed immediately by the enrichments. After several transfers of the enrichments, a stable consortium of at least three bacterial types was obtained. The predominant bacterium was a non-motile, gram-negative coccus. This stable consortium was able to remove chlorinated methanes, ethanes, and ethylenes when grown with methane and oxygen in the headspace. Methane was required for the removals to be observed. Acetylene inhibited the removals, which further suggests the involvement of methanotrophs. Benzene and toluene were removed by the mixed culture with or without methane in the headspace. Fatty acid analysis of the mixed culture resulted in a profile that indicated that the predominant organism was a type II methanotroph. This study provides further evidence that methanotrophic bacteria are capable of cometabolizing a wide range of chlorinated methanes, ethanes, and ethylenes.     

Assimilation of methane and inorganic carbon by microbial communities mediating the anaerobic oxidation of methane

The anaerobic oxidation of methane (AOM) is a major sink for methane on Earth and is performed by consortia of methanotrophic archaea (ANME) and sulfate-reducing bacteria (SRB). Here we present a comparative study using in vitro stable isotope probing to examine methane and carbon dioxide assimilation into microbial biomass. Three sediment types comprising different methane-oxidizing communities (ANME-1 and -2 mixture from the Black Sea, ANME-2a from Hydrate Ridge and ANME-2c from the Gullfaks oil field) were incubated in replicate flow-through systems with methane-enriched anaerobic seawater medium for 5–6 months amended with either ¹³CH₄ or H¹³CO₃^-. In all three sediment types methane was anaerobically oxidized in a 1:1 stoichiometric ratio compared with sulfate reduction. Similar amounts of ¹³CH₄ or ¹³CO₂ were assimilated into characteristic archaeal lipids, indicating a direct assimilation of both carbon sources into ANME biomass. Specific bacterial fatty acids assigned to the partner SRB were almost exclusively labelled by ¹³CO₂, but only in the presence of methane as energy source and not during control incubations without methane. This indicates an autotrophic growth of the ANME-associated SRB and supports previous hypotheses of an electron shuttle between the consortium partners. Carbon assimilation efficiencies of the methanotrophic consortia were low, with only 0.25–1.3 mol% of the methane oxidized.     

Impact of nitrate-mediated microbial control of souring in oil reservoirs on the extent of corrosion

The effect of microbial control of souring on the extent of corrosion was studied in a model system consisting of pure cultures of the nitrate-reducing, sulfide-oxidizing bacterium (NR-SOB) Thiomicrospira sp. strain CVO and the sulfate-reducing bacterium (SRB) Desulfovibrio sp. strain Lac6, as well as in an SRB consortium enriched from produced water from a Canadian oil reservoir. The average corrosion rate induced by the SRB consortium (1.4 g x m(-2) x day(-1)) was faster than that observed in the presence of strain Lac6 (0.2 g x m(-2) x day(-1)). Examination of the metallic coupons at the end of the tests indicated a uniform corrosion in both cases. Addition of CVO and 10 mM nitrate to a fully grown culture of Lac6 or the SRB consortium led to complete removal of sulfide from the system and a significant increase in the population of CVO, as determined by reverse sample genome probing. In the case of the SRB consortium addition of just nitrate (10 mM) had a similar effect. When grown in the absence of nitrate, the consortium was dominated by Desulfovibrio sp. strains Lac15 and Lac29, while growth in the presence of nitrate led to dominance of Desulfovibrio sp. strain Lac3. The addition of CVO and nitrate to the Lac6 culture or nitrate to the SRB consortium accelerated the average corrosion rate to 1.5 and 2.9 g x m(-2) x day(-1), respectively. Localized corrosion and the occurrence of pitting were apparent in both cases. Although the sulfide concentration (0.5-7 mM) had little effect on corrosion rates, a clear increase of the corrosion rate with increasing nitrate concentration was observed in experiments conducted with consortia enriched from produced water.     

Microbiological and biogeochemical processes in a pockmark of the Gdansk depression,Baltic Sea

Comprehensive microbiological and biogeochemical investigation of a pockmark within one of the sites of gas-saturated sediments in the Gdansk depression, Baltic Sea was carried out during the 87th voyage of the Professor Shtokman research vessel. Methane content in the near-bottom water and in the underlying sediments indicates stable methane flow from the sediment into the water. In the 10-m water layer above the pockmark, apart from methane anomalies, elevated numbers of microorganisms and enhanced rates of dark CO₂ fixation (up to 1.15 µmol C/(l day)) and methane oxidation (up to 2.14 nmol CH₄/(l day)) were revealed. Lightened isotopic composition of suspended organic matter also indicates high activity of the near-bottom microbial community. Compared to the background stations, methane content in pockmark sediments increased sharply from the surface to 40–60 ml/dm³ in the 20–30 cm horizon. High rates of bacterial sulfate reduction (SR) were detected throughout the core (0–40 cm); the maximum of 74 µmol S/(dm³ day) was located in subsurface horizons (15–20 cm). The highest rates of anaerobic methane oxidation (AMO), up to 80 µmol/dm³ day), were detected in the same horizon. Good coincidence of the AMO and SR profiles with stoichiometry close to 1: 1 is evidence in favor of a close relation between these processes performed by a consortium of methanotrophic archaea and sulfate-reducing bacteria. Methane isotopic composition in subsurface sediments of the pockmark (from ?53.0 to ?56.5‰) does not rule out the presence of methane other than the biogenic methane from the deep horizons of the sedimentary cover.     

含有甲烷氧化菌的混合菌群特性研究

为获得高效甲烷氧化微生物体系,从农业土壤中采样,以甲烷作为唯一碳源进行好氧选择性传代培养,得到生长性能稳定、生长优于Methylosinus trichosporium OB3b纯培养的具有甲烷单加氧酶(Methane Monooxygenase,MMO)活性甲烷氧化混合菌。利用MMO的共代谢特性,分别以苯酚和环氧丙烷作为目标对象,考察该混合菌对有机污染物的降解及用于生产有用化学物质的催化特性。结果表明,所得混合菌具有高效降解苯酚能力,对初始浓度为600mg/L的苯酚,经过11h培养,苯酚降解率可达99%。另外,以该混合菌为催化剂可以实现丙烯氧化生产环氧丙烷。通过降低磷酸盐浓度可以有效提高环氧丙烷的积累浓度,最大可至5mmol/L。此外,采用纯种分离方法结合PCR扩增、16SrRNA和MMO功能基因分析技术对混合菌群结构进行解析。结果表明,该混合菌群由Ⅱ型甲烷氧化菌及其它至少4种非甲烷氧化菌组成,它们分别属于Methylosinus trichosporium和Acinetobacter junii、Cupriavidusme tallidurans、Comamonas testosteroni和Stenotrophomonas maltophilia。采用PCR方法从混合菌及纯化菌株M.trichosporiums Y9总DNA中都能扩增得到mmoB、mmoX和pmoA基因片段,表明该甲烷氧化菌同时具有sMMO和pMMO两种形式的MMO。通过对从甲烷氧化混合菌中分离纯化得到的甲烷氧化菌进行PCR产物测序,结果发现其与Methylosinus trichosporium的同源性为99.9%。     

Zoonotic bacterial populations, gut fermentation characteristics and methane production in feedlot steers during oral nitroethane treatment and after the feeding of an experimental chlorate product

Nitroethane inhibits the growth of certain zoonotic pathogens such as Campylobacter and Salmonella spp., foodborne pathogens estimated to cause millions of human infections each year, and enhances the Salmonella- and Escherichia coli-killing effect of an experimental chlorate product being developed as a feed additive to kill these bacteria immediately pre-harvest. Limited studies have shown that nitroethane inhibits ruminal methane production, which represents a loss of 2-12% of the host's gross energy intake and contributes to global warming and destruction of the ozone layer. The present study was conducted to assess the effects of 14-day oral nitroethane administration, 0 (0X), 80 (1X) or 160 (2X)mg nitroethane/kg body weight per day on ruminal and fecal E. coli and Campylobacter, ruminal and fecal methane-producing and nitroethane-reducing activity, whole animal methane emissions, and ruminal and fecal fermentation balance in Holstein steers (n=6 per treatment) averaging 403+/-26 (SD) kg BW. An experimental chlorate product was fed the day following the last nitroethane administration to determine effects on E. coli and Campylobacter. The experimental chlorate product decreased (P<0.001) fecal, but not ruminal (P>0.05) E. coli concentrations by 1000- and 10-fold by 24 and 48 h, respectively, after chlorate feeding when compared to pre-treatment concentrations (>5.7 log(10) colony forming units/g). No effects (P>0.05) of nitroethane or the experimental chlorate product were observed on fecal Campylobacter concentrations; Campylobacter were not recovered from ruminal contents. Nitroethane treatment decreased (P<0.01) ruminal (8.46, 7.91 and 4.74+/-0.78 micromol/g/h) and fecal (3.90, 1.36 and 1.38+/-0.50 micromol/g/h) methane-producing activity for treatments 0X, 1X and 2X, respectively. Administration of nitroethane increased (P<0.001) nitroethane-reducing activity in ruminal, but not fecal samples. Day of study affected ruminal (P<0.0001) but not fecal (P>0.05) methane-producing and nitroethane-reducing activities (P<0.01); treatment by day interactions were not observed (P>0.05). Ruminal accumulations of acetate decreased (P<0.05) in 2X-treated steers when compared with 0X- and 1X-treated steers, but no effect (P>0.05) of nitroethane was observed on propionate, butyrate or the acetate to propionate ratio. Whole animal methane emissions, expressed as L/day or as a proportion of gross energy intake (%GEI), were unaffected by nitroethane treatment (P>0.05), and were not correlated (P>0.05) with ruminal methane-producing activity. These results demonstrate that oral nitroethane administration reduces ruminal methane-producing activity but suggest that a microbial adaptation, likely due to an in situ enrichment of ruminal nitroethane-reducing bacteria, may cause depletion of nitroethane, at least at the 1X administration dose, to concentrations too low to be effective. Further research is warranted to determine if the optimization of dosage of nitroethane or related nitrocompouds can maintain the enteropathogen control and anti-methanogen effect in fed steers.     

Mass balance and life cycle assessment of biodiesel from microalgae incorporated with nutrient recycling options and technology uncertainties

This article presents mass balances and a detailed life cycle assessment (LCA) for energy and greenhouse gases (GHGs) of a simulated microalgae biodiesel production system. Key parameters of the system include biomass productivity of 16 and 25 g m^?2 day^?1 and lipid content of algae of 40% and 25% for low and normal nitrogen conditions respectively. Based on an oil extraction efficiency from wet biomass of 73.6% and methane yields from anaerobically digested lipid‐extracted biomass of 0.31 to 0.34 l per gram of volatile solids, the mass balance shows that recycling growth media and recovering nutrients from residual biomass through anaerobic digestion can reduce the total demand for nitrogen by 66% and phosphorus by 90%. Freshwater requirements can be reduced by 89% by recirculating growth media, and carbon requirements reduced by 40% by recycling CO₂ from biogas combustion, for normal nitrogen conditions. A variety of technology options for each step of the production process and allocation methods for coproducts used outside the production system are evaluated using LCA. Extensive sensitivity and scenario analysis is also performed to provide better understanding of uncertainty associated with results. The best performing scenario consists of normal nitrogen cultivation conditions, bioflocculation and dissolved air flotation for harvesting, centrifugation for dewatering, wet extraction with hexane, transesterification for biodiesel production, and anaerobic digestion of biomass residual, which generates biogas used in a combined heat and power unit for energy recovery. This combination of technologies and operating conditions results in life cycle energy requirements and GHG emissions of 1.02 MJ and 71 g CO₂‐equivalent per MJ of biodiesel, with cultivation and oil extraction dominating energy use and emissions. Thus, even under optimistic conditions, the near‐term performance of this biofuel pathway does not achieve the significant reductions in life cycle GHG emissions hoped for from second‐generation biofuel feedstocks.     

Microbiological Processes in a High-Temperature Oil Field

Thermophilic sulfate-reducing bacteria (SRB) oxidizing lactate, butyrate, and C₁₂–C₁₆ n-alkanes of oil at a temperature of 90°C were isolated from samples of water and oil originating from oil reservoirs of the White Tiger high-temperature oil field (Vietnam). At the same time, no thermophiles were detected in the injected seawater, which contained mesophilic microorganisms and was the site of low-temperature processes of sulfate reduction and methanogenesis. Thermophilic SRB were also found in samples of liquid taken from various engineering reservoirs used for oil storage, treatment, and transportation. These samples also contained mesophilic SRB, methanogens, aerobic oil-oxidizing bacteria, and heterotrophs. Rates of bacterial production of hydrogen sulfide varied from 0.11 to 2069.63 at 30°C and from 1.18 to 173.86 at 70°C g S/(l day); and those of methane production, varied from 58.4 to 100 629.8 nl CH₄/(l day) (at 30°C). The sulfur isotopic compositions of sulfates contained in reservoir waters and of hydrogen sulfide of the accompanying gas indicate that bacterial sulfate reduction might be effective in the depth of the oil field.     

Characterization and optimization of biosurfactants produced by Acinetobacter baylyi ZJ2 isolated from crude oil-contaminated soil sample toward microbial enhanced oil recovery applications

The present work aims to investigate the surface activity of the biosurfactant produced by Acinetobacter baylyi ZJ2 isolated from crude oil-contaminated soil sample in China and evaluate its potential application in microbial enhanced oil recovery. The biosurfactant produced by A. baylyi ZJ2 was identified as lipopeptide based on thin-layer chromatography, Fourier transform infrared spectroscopy and nuclear magnetic resonance techniques. This biosurfactant could reduce the surface tension of water from 65 mN/m to 35 mN/m, and interfacial tension against oil from 45 mN/m to 15 mN/m. Moreover, surface activity stability results showed that this biosurfactant was effective when the salinity was lower than 8% and the pH value was 4–9, and it was especially stable when the salinity was lower than 4% and pH was 6–7. Based on the result of gas chromatography, there was a decrease in heavy components and an increase in light components, which indicated that A. baylyi ZJ2 exhibited the biodegradability on the heavy components of crude oil. Furthermore, the ability of recovering oil from oil-saturated core showed that nearly 28% additional residual oil was displaced after water flooding. The lipopeptide biosurfactant produced by A. baylyi ZJ2 presented a great potential application in microbial enhanced oil recovery process, owing its good surface activity and satisfying degradation ability to crude oil.