Inhibition of Acetoclastic Methanogenesis in Crude Oil- and Creosote-Contaminated Groundwater

Results from a series of studies of methanogenic processes in crude oil- and creosote-contaminated aquifers indicate that acetoclastic methanogenesis is inhibited near non-aqueous sources. At a crude oil-contaminated site, numbers of acetoclastic methanogens found close to crude oil were one hundred times fewer than those of hydrogen- and formate-utilizing methanogens. In laboratory toxicity assays, crude oil collected from the site inhibited methane production from acetate but not from formate or hydrogen. Toxicity assays with aqueous creosote extract completely inhibited acetate utilization over the range of tested dilutions but only mildly affected formate and hydrogen utilization. The combined results from the laboratory and field studies suggest that in methanogenic contaminated aquifers, inhibition of acetoclastic methanogenesis may lead to a buildup of acetate relative to dissolved organic carbon.     

Inhibition of Acetoclastic Methanogenesis in Crude Oil- and Creosote-Contaminated Groundwater

Results from a series of studies of methanogenic processes in crude oil- and creosote-contaminated aquifers indicate that acetoclastic methanogenesis is inhibited near non-aqueous sources. At a crude oil-contaminated site, numbers of acetoclastic methanogens found close to crude oil were one hundred times fewer than those of hydrogen- and formate-utilizing methanogens. In laboratory toxicity assays, crude oil collected from the site inhibited methane production from acetate but not from formate or hydrogen. Toxicity assays with aqueous creosote extract completely inhibited acetate utilization over the range of tested dilutions but only mildly affected formate and hydrogen utilization. The combined results from the laboratory and field studies suggest that in methanogenic contaminated aquifers, inhibition of acetoclastic methanogenesis may lead to a buildup of acetate relative to dissolved organic carbon.     

Inhibition of Acetoclastic Methanogenesis in Crude Oil- and Creosote-Contaminated Groundwater

Results from a series of studies of methanogenic processes in crude oil- and creosote-contaminated aquifers indicate that acetoclastic methanogenesis is inhibited near non-aqueous sources. At a crude oil-contaminated site, numbers of acetoclastic methanogens found close to crude oil were one hundred times fewer than those of hydrogen- and formate-utilizing methanogens. In laboratory toxicity assays, crude oil collected from the site inhibited methane production from acetate but not from formate or hydrogen. Toxicity assays with aqueous creosote extract completely inhibited acetate utilization over the range of tested dilutions but only mildly affected formate and hydrogen utilization. The combined results from the laboratory and field studies suggest that in methanogenic contaminated aquifers, inhibition of acetoclastic methanogenesis may lead to a buildup of acetate relative to dissolved organic carbon.     

Isolation and characterization of Methanoculleus receptaculi sp. nov. from Shengli oil field, China

Three strictly anaerobic, thermophilic methanogens (ZC-2T, ZC-3 and ZC-6) were isolated from Shengli oil field, China. The 16S rRNA gene sequences of the three strains were nearly identical, possessing > 99.8% sequence similarity. They also possessed high sequence similarity, 97.4%, to Methanoculleus palmolei strain INSLUZ(T) (97.4% and 97.5%, respectively), indicating that they represented a novel species within the genus Methanoculleus. Cells of strain ZC-2T were nonmotile cocci, 0.8-1.7 microm in diameter, and always occurred singly or in pairs. The three strains used H2/CO2 or sodium formate as substrates for methanogenesis but not sodium acetate, trimethylamine, monomethylamine, ethanol, dimethyl sulfide, isopropanol, isobutanol, butan-2-ol or H2/CO. Optimum growth of strain ZC-2T occurred in the presence of 0.2 M NaCl, pH 7.5-7.8 and temperature 50-55 degrees C with a specific growth rate of 0.084 h(-1). The mol% G+C content of the genomic DNA was 55.2 mol%. Based on these phenotypic and phylogenetic characteristics, strains ZC-2T, ZC-3 and ZC-6 are proposed to represent a novel species in the genus Methanoculleus and named Methanoculleus receptaculi sp. nov. The type strain is ZC-2T (CGMCC 1.5087T=DSM 18860T).     

芦岭煤田微生物群落结构和生物成因气的产甲烷类型研究

【目的】揭示芦岭煤田微生物群落组成,并分析其潜在的产甲烷类型及产甲烷途径。【方法】采集芦岭煤田的煤层气样品和产出水样品,分别分析样品的地球化学性质特征;利用Illumina HiSeq高通量测序技术分析产出水中的微生物群落结构;采用添加不同底物的厌氧培养实验进一步证实芦岭煤田生物成因气的产甲烷类型。【结果】该地区煤层气为生物成因和热成因的混合成因气;古菌16S rRNA基因分析表明在产出水中含有乙酸营养型、氢营养型和甲基营养型的产甲烷菌。丰度较高的细菌具有降解煤中芳香族和纤维素衍生化合物的潜力。厌氧富集培养结果表明,添加乙酸盐、甲酸盐、H2+CO2为底物的矿井水样均有明显的甲烷产生。【结论】芦岭煤田具有丰富的生物多样性,该地区同时存在三种产甲烷类型。本研究为利用微生物技术提高煤层气的采收率,实现煤层气的可持续开采提供科学依据。     

产甲烷菌研究进展

产甲烷菌是重要的环境微生物,在自然界的碳素循环中起重要作用。迄今已有5种产甲烷菌基因组测序完成。基因组信息使人们对产甲烷茵的细胞结构、进化、代谢及环境适应性有了更深的理解。目前已知的甲烷生物合成途径有3种,它们以乙酸、甲基化合物、氢/二氧化碳为起始,通过不同的反应途径都形成了甲基辅酶M,在甲基辅酶M还原酶的催化下最终形成甲烷。     

Prepared Bed Land Treatment of Soils Containing Diesel and Crude Oil Hydrocarbons

An ex situ, field-scale, prepared bed land treatment unit (LTU) was used to bio-remediate soils containing petroleum hydrocarbons. Two soils were treated in side-by-side units to compare performance: (1) a clayey silt containing crude oil hydrocarbons from releases 30 to 40 years ago and (2) a silty sand containing diesel fuel hydrocarbons from a leak about three years prior to the bioremediation. The effectiveness of the bioremediation in the LTU was evaluated over a period of 18 months. The results indicated that: (1) prepared bed bioremediation reduced the hydrocarbon concentration, mobility, and relative toxicity in the soil with the diesel fuel, and (2) chemical bioavailability appeared to limit bioremediation of the soil containing the crude oil hydrocarbons. Although the soils containing the crude oil hydrocarbons contained an average of 10,000?mg TPH/kg dry soil, these soils had limited hydrocarbon availability, nontoxic conditions, and low potential for chemical migration. For the soils containing the diesel fuel, active prepared bed bioremediation of about 15 weeks was adequate to reach an environmentally acceptable endpoint. At that time, there was little further TPH loss, no Microtox^TM toxicity, and limited hydrocarbon mobility.     

Conversion and toxicity characteristics of formaldehyde in acetoclastic methanogenic sludge

An unadapted mixed methanogenic sludge transformed formaldehyde into methanol and formate. The methanol to formate ratio obtained was 1:1. Formaldehyde conversion proceeded without any lag phase, suggesting the constitutive character of the formaldehyde conversion enzymes involved. Because the rate of formaldehyde conversion declined at increased formaldehyde additions, we hypothesized that some enzymes and/or cofactors might become denatured as a result of the excess of formaldehyde. Furthermore, formaldehyde was found to be toxic to acetoclastic methanogenesis in a dual character. Formaldehyde toxicity was partly reversible because once the formaldehyde concentration was extremely low or virtually removed from the system, the methane production rate was partially recovered. Because the degree of this recovery was not complete, we conclude that formaldehyde toxicity was partly irreversible as well. The irreversible toxicity likely can be attributed to biomass formaldehyde-related decay. Independent of the mode of formaldehyde addition (i.e., slug or continuous), the irreversible toxicity was dependent on the total amount of formaldehyde added to the system. This finding suggests that to treat formaldehyde-containing waste streams, a balance between formaldehyde-related decay and biomass growth should be attained.     

Enhancing the Removal of Sorbed Crude Oil from Soil Through Multiple Oxidation Steps in Stepwise Fenton Processes

The stepwise Fenton oxidation process, in which hydrogen peroxide (H₂O₂) is added in a step-by-step manner instead of at the beginning, can achieve better sorbed crude oil removal effects. The results showed that if a high ratio of sorbed total petroleum hydrocarbon (TPH) was present in soil samples S1 (100%, initial TPH: 10,009 mg/kg) and S2 (94.2%, initial TPH: 4850 mg/kg), the TPH was oxidized in each step. In addition, the total TPH removal efficiency was 49.6% compared with the 27.9% achieved in conventional Fenton oxidation in which all H₂O₂ was added at the beginning. Nevertheless, when the ratio of sorbed TPH in the soil sample S3 was low (45.3%, initial TPH: 2850 mg/kg), the TPH removal efficiency was 18.9%, which was slightly higher than 18.2% achieved in the conventional Fenton process because if the sorbed TPH concentration was low, the sorbed TPH was mainly removed in the first step. The second and the third step resulted in long-chain alkanes entering the aqueous phase rather than removing them from the soil, which posed environmental risk. Therefore, it is clear that stepwise Fenton oxidation could improve sorbed TPH removal efficiency when the sorbed TPH concentration in the soil is high.     

海绵中石油降解微生物的分离筛选及降解特性研究

从大连湾原油污染海域生长的海绵中分离筛选到一株原油降解菌OA58,根据其生长形态、培养特征、16S rRNA序列相似性比对分析和生化指标检测,初步鉴定为脱叶链霉菌（Streptomyces exfoliatus）。同时,考察了该链霉菌对原油的降解效果,OA58能以原油为唯一碳源生长,在人工海水培养基中,14 d内对原油（初始浓度为1 g/L）的平均降解率为83%,是一株具有开发潜力的原油降解放线菌。     

石油集输系统中微生物群落结构研究

采用16SrRNA基因克隆一变性梯度凝胶电泳分析方法研究了石油集输系统原油和油田产水中的微生物群落结构。变性梯度凝胶电泳图谱显示：油田产水中微生物群落远比原油中的菌群丰富。所有的油田水样和原油样本中都存在与Ochrobactrum sp．和Stenotrophomonas sp．相关的细菌;原油样本中检测出与Burkholderia sp．、Brevundimonas sp．和Propionibacterium sp．相关的细菌,而这些细菌在油田水样中未检出;在油田水样中检出与Hippea sp．、Acidovorax sp．、Arcobacter sp．、Pseudomonas sp．、Thiomicrospira sp．、Brevibacterium sp．、Tissierella sp．和Peptostreptococcus sp．相关的细菌,而这些细菌在原油样本中未检出。用古细菌特异性引物进行检测发现在油田水样中存在与Methanomicrobials和Methanosarcinales相关的产甲烷菌,而这些细菌在原油样本中未检出。在石油集输过程中,油田水样和原油中微生物群落的相似性分别为83．3％和88．2％,说明微生物群落结构较为稳定。     

Feasibility Study of Performing an Life Cycle Assessment on Crude Palm Oil Production in Malaysia (9 pp)

Background, Goal and Scope The palm oil industry is one of the leading industries in Malaysia. With a yearly production of more than 13 million tons of crude palm oil (CPO) and plantations covering 11% of the Malaysian land area it is an industry to be reckoned with, also when it comes to environmental impacts. One way to describe and present the environmental impacts is through a life cycle assessment, LCA. This assessment aims to introduce the concept of LCA and perform a screening LCA on crude palm oil production in Malaysia including the stages of plantation, transport and milling. The assessment is largely based on general data and is thus meant to function as an indication of the environmental threads posed by CPO production and as a guideline to CPO producers and local universities on how to perform an LCA on a palm oil scenario. Due to the general data background the results of this report should not be quoted directly for decision making. The Functional Unit, to which all masses and emissions in this assessment have been adjusted, is the production of 1000 kg of CPO in Malaysia. Method Initially an overview of palm oil production was obtained and the outlines and borders of the assessment were determined along with the specific goal and scope of the assessment. The data for the assessment was collected from three different sources: - 1. Earlier studies and statistics on palm oil production in Malaysia - 2. Studies on similar processes, when palm oil related processes were not available - 3. General data from the SimaPro 5 database - The European Eco-Indicator 99 method and European databases included in the LCA software SimaPro 5 have been used for the impact calculations. Results and Discussion The impact processes related to the plantation are the on-site energy use (mainly diesel) and the production of artificial fertilizer. Pesticide use contributes a minor impact due to widely used integrated biological poet management. For transportation the only impact is from combustion of diesel and at the mill the boiler is the sole significant contributor – positively through electricity production and negatively by emissions from the boiler. Impacts from POME (Palm Oil Mill Effluent) are not dealt with in the main assessment, but touched upon in alternative scenarios. The results clearly show that fertilizer production is the most polluting process in the system followed by transportation and the boiler emissions at a tie. The most significant impacts from the system are respiratory inorganics and depletion of fossil fuels, of which the boiler emission is the main responsible for the prior and fertilizer production and transportation are responsible for the latter. It is also evident from the results that crude palm oil production is a significant environmental impact generator in Malaysia due to the vast production quantities. Alternative scenarios revealed that there are significant impact savings to be made by introduction of environmental investments, both regarding the overall impacts and in particularly regarding CO2 emissions. Conclusion A screening LCA was successfully conducted on the Malaysian crude palm oil production thus promising potentials for the palm oil industry to conduct their own inventories and assessments using specific company data. Crude palm oil production in Malaysia is responsible for app. 3.5% of the total environmental impacts in the country and must thus be given attention to reduce impacts. Alternatives such as optimized use of organic fertilizer, environmentally friendlier artificial fertilizer production, rail transport, approved filters at the mill boiler stack and biogas harvest from POME digestion must thus be promoted in the industry. Recommendation . The Malaysian palm oil industry should take steps towards introducing LCA. Exhaustive inventories are likely to open the eyes of many companies towards implementing environmental investments and improve the international competitiveness. In order to retrieve results with a greater accuracy in the future, databases must be created containing life cycle data from Malaysian scenarios and normalization and weighting factors must be designed to reflect Malaysian conditions. The Malaysian authorities must create incentives through increased tariffs on electricity and diesel and/or financial support for cleaner technology investments.     

原油和消油剂对鱼类毒性的研究进展

原油对鱼类的毒性主要来自其水溶性成分,可导致鱼卵死亡或发育畸形,并造成鱼类麻痹、发炎、粘膜受损和死亡。消油剂的使用在分散了油膜的同时也带来二次污染,其活性剂对鱼类产生新的毒性影响。消油剂将原油分散为乳化颗粒并能进入鱼体内,加大了原油的毒性并延长其作用时间。综述了原油和消油剂对鱼类毒性的研究进展,展望了鱼类在相关毒性试验领域的应用及发展方向。     

Variation in Petroleum Hydrocarbon Chain Lengths with Depth at a Former Crude Oil and Natural Gas ProductionFacility

An investigation was performed at a former crude oil and natural gas production facility to evaluate whether releases from the product flowlines, gathering lines or water injection lines had impacted soil beneath the site. Thirty-six trenches were initially excavated and sampled beneath the former piping runs to a maximum depth of 6?m. After the trenching investigation, nine soil boreholes were advanced and sampled to a depth of approximately 18?m to further delineate the lateral and vertical extent of impacted soil. Soil samples collected from the trenches and boreholes were analyzed for total petroleum hydrocarbons (TPH) in accordance with ASTM Method 2887. The results of the investigation indicated that TPH impacted soil was present within several areas of the 40-ha site. The petroleum hydrocarbons generally had chain lengths ranging from C₆ to C₃₅, characteristic of light crude oil. The impacted soil also contained condensate, the volatile portion of crude oil. Condensate consists of short-chain hydrocarbons (C₁ to C₁₂) and is characterized by low levels of aromatic volatile organic compounds (VOCs). The condensate typically was more prevalent at depths below 4.5?m than the less volatile, longer chain length hydrocarbons. Statistical analysis of TPH data collected during subsequent excavation activities showed that the mean percentage of condensate was significantly greater at depths below 4.5?m than in shallower samples. In contrast, the mean percentage of TPH compounds in the diesel range (C₁₄ to C₂₃) was significantly greater in samples collected at depths above 4.5?m. The difference in the mean percentage of heavier hydrocarbons (C₂₄ to C₄₄+) with depth was not statistically significant.     

Microbial Community Composition in Crude Oils and Asphalts from the Kurdistan Region of Iraq

Abstract

To identify hydrocarbon-degrading microorganisms contributing to the formation of heavy oil we investigated the microbial community composition in different types of crude oils from oil-production facilities and in crude oil and asphalt from different natural seeps from the Kurdistan Region of Iraq (KRI). Crude oils from five out of six production facilities did not contain microorganisms detectable by 16S rRNA gene PCR amplicon sequencing likely reflecting a low microbial abundance in these samples. Crude oil and asphalt from the natural seeps hosted diverse microbial communities. The same phylotypes of uncultivated Deferribacteres and Thermodesulfobacteraceae were predominant community members across crude oils and asphalts from separate geographical locations. Soils surrounding seeps did not contain these phylotypes suggesting that they originate from the subsurface and although they seem commonly detected in hydrocarbon-rich environments their role in hydrocarbon-degradation is unknown. GC-MS analyses showed that mainly aromatic hydrocarbons were present in the crude oil and asphalt and that they were undergoing biodegradation - likely with sulfate and nitrate as terminal oxidants. In agreement, only bssA gene, but not assA gene-carrying microorganisms were detectable in the analyzed sampled. Overall our study identified several abundant uncultivated taxa with likely roles in transformation of nitrate, sulfate and hydrocarbons.     

Biodegradation of Aliphatic and Aromatic Fractions of Heavy Crude Oil-Contaminated Soil: A Pilot Study

Crude oil is one of the major contaminants of soil and water. Biodegradation is a good option for reducing oil contamination in soil. Soil fertilizers can help the microorganisms biodegrade oil with higher efficiency. In this study, the effect of chemical fertilizers and animal manure on crude oil reduction was compared and the reduction of the aliphatic and aromatic fractions was studied during 8 months. The higher reduction of crude oil (54%) was observed in tests containing chemical fertilizers and wood chips, followed by those containing animal manure (46%). The reduction of aliphatic and aromatic fractions was also higher in fertilized tests. Our results showed that in the experiments with mixed chemical fertilizers and wood chips, the reduction of crude oil and its aliphatic and aromatic fractions was higher than the experiments with only chemical fertilizers and those with animal manure. The reductions in the experiments with animal manure, however, were higher than those with only chemical fertilizers.     

Inoculants and Biodegradation of Crude Oil Floating on Marsh Sediments

The efficacy of ten commercial bioremediation products in enhancing the biodegradation of crude oil was investigated in the laboratory at 10 or 30a°C for 90 d with and without supplemental nitrogen and phosphorus. Oil was added to a 1-cm layer of water covering sediments from a salt marsh. The products did not increase the numbers of hydrocarbon-degrading microorganisms in water and sediments but did increase heterotrophic populations at 21 d. Some bioremediation products more than doubled the quantity of hydrocarbons degraded in 45 d at 10°C. At 30°C, no product increased degradation compared to the fertilized control in which 70% of the added hydrocarbons were degraded. Two products increased the percentage of hydrocarbons degraded from 42% to approximately 65% in 45 d at 30°C when supplemental fertilizer was not provided. The hydrocarbon concentration was not significantly reduced between 45 and 90 d for most product treatments at either temperature. At 10°C, products seemed to have the greatest potential for enhancing oil bioremediation compared to the control.     

Effects of Light Crude Oil Contamination on the Physical and Mechanical Properties of Fine Sand

The use of oil-contaminated sand in building and construction is now being considered as an alternative and cost-effective way to minimize its adverse effect on the environment. To achieve this, the effect of oil contamination on the important mechanical properties of sand should be investigated first. This study investigated the effect of petroleum-derived contaminants on the water absorption, permeability, cohesion, friction angle, and shear strength of fine sand. Contaminated samples were prepared by mixing fine sand with different percentages of light crude oil (0 to 20%). The results indicated that the water absorption of fine sand decreases with an increase in crude oil. An increase in the cohesion was observed for sand with up to 1% of oil contamination, after which the cohesion began to decrease, which also results in the reduction in the permeability. A slight reduction in the friction angle was found for oil-contaminated fine sand. At a low normal stress of 50 kPa, as the percentage of light crude oil increased, the shear strength increased up to 1% of oil contamination and then it decreased. These results provided useful information on how oil-contaminated sand can be used safely and effectively in building and construction.     

Isolation and Characterization of Novel Hydrocarbon-Degrading Euryhaline Consortia from Crude Oil and Mangrove Sediments

Two novel and versatile bacterial consortia were developed for the biodegradation of hydrocarbons. They were isolated from crude oil from the Cormorant Field in the North Sea (MPD-7) and from sediment associated with mangrove roots (MPD-M). The bacterial consortia were able to degrade both aliphatic and aromatic hydrocarbons in crude oils very effectively in seawater (35 g/L NaCl) and synthetic media containing 0 to 100 g/L NaCl (1.7 M). Salinities over twice that of normal seawater decreased the biodegradation rates. However, even at the highest salinity biodegradation was significant. Ratios of nC17 to pristane and nC18 to phytane were significantly lowered across the range of salinity. The lowest values were at 0 and 20 g/L (0.34 M). Phytane was degraded in preference to pristane. The degradation of these compounds was constant over the salinity range, with evidence of a slight increase for consortium MPD-M with increasing salinity. In general, the consortium isolated from mangrove root sediments was more efficient in metabolizing North Sea crude oil than the consortium isolated from Cormorant crude oil. The 5 strains that comprise MPD-M have been tentatively identified as species of the genera Marinobacter, Bacillus, and Erwinia. This is the first report of hydrocarbon-degrading consortia isolated from crude oil and mangrove sediments that are capable of treating oily wastes over such a wide range of salinity. Received June 30, 1999; accepted May 29, 2000.     

Oil Palm Empty Fruit Bunch and Sugarcane Bagasse Enhance the Bioremediation of Soil Artificially Polluted by Crude Oil

Contamination of soil by petroleum hydrocarbons is becoming prevalent in Malaysia. Infiltration of soil contamination into groundwater poses a great threat to the ecosystem and human health. Bioremediation can occur naturally or can be enhanced with supplementation of microorganisms and fertilizers. However, fertilizers are expensive and therefore alternative nutrient-rich biomaterials are required. In this study, two organic wastes from agricultural industry (i.e., sugarcane bagasse and oil palm empty fruit bunch) were investigated for possible enhanced bioremediation of soil contaminated with Tapis crude oil. Two bacterial strains isolated and characterized previously (i.e., Pseudomonas aeruginosa UKMP-14T and Acinetobacter baumannii UKMP-12T) were used in this study. Sugarcane bagasse (5% and 15%, w/w) and oil palm empty fruit bunch (20%, w/w) were mixed with soil (500 g) spiked with Tapis crude oil (3%, v/w). The treated soils as well as controls were incubated for 20 days under controlled conditions. Sampling was carried out every four days to measure the number of bacterial colonies (CFU/g) and to determine the percentage of oil degradation by gas chromatography. The two biostimulating agents were able to maintain the soil moisture holding capacity, pH, and temperature at 38-40% volumetric moisture content (VMC), 7.0, and 29–30°C; respectively. The growth of bacteria consortium after 20 days in the treatment with sugarcane bagasse and oil palm empty fruit bunch had increased to 10.3 CFU/g and 9.5 CFU/g, respectively. The percentage of hydrocarbon degradation was higher in the soil amended with sugarcane bagasse (100%) when compared to that of oil palm empty fruit bunch (97%) after 20 days. Our results demonstrated the potential of sugarcane bagasse and oil palm empty fruit bunch as good substrates for enhanced bioremediation of soil contaminated with petroleum crude oil.