Assessment of the Integral Toxicity of Aquatic Medium Contamination with Oil and Oil Products Using Bacterial Tests

A biotest kit was used to assess the integral toxicity level of aquatic environment contamination with oil and oil products. The integral toxicity dynamics were also monitored during biodegradation of oil and oil products by an association of oil-degrading strains, including Acinetobacter sp., Mycobacterium flavescens, and Rhodococcussp. The following bacterial tests were used: the bioluminescence (BL) test based on Photobacterium leiognathi; electroorientation (EO), optoosmotic (OO), and growth tests; and the reducing activity (RA) test based on an Agrobacterium radiobacter culture. No significant increase in the integral toxicity level of the aquatic medium was observed when diesel fuel and kerosene contamination had been subjected to biodegradation. Although rapid biotests (EO, OO, RA, and BL) detected a pronounced increase in the integral toxicity of the aquatic environment, long-term growth biotests revealed no statistically significant increase in the toxicity level.     

Plant biotests of soil and water, polluted with petroleum and petroleum products

Reactiona of higher plants (mustard, oat, rye, salad, dill and barley) and microalgae (Euglena gracilis) on the contamination of soil and water with petroleum and oil products was studied. The germination of seeds was analyzed. The length of sprouts, dry biomass and length of plant roots, as well as the optical density of micro-algal broth culture were determined. Negative effects of soil and water contamination with petroleum and oil products on plant and microalgal parameters examined was shown. After biological destruction of contaminants by an association of destructor strains (Acinetobacter sp., Mycobacterium flavescens and Rhodoccocus sp.), the toxicity of contaminated mediums decreased. The data suggest that the integral toxicity of soil and water contaminated with petroleum and oil products and toxicity change during biodestruction of these pollutants can be analyzed by using plant test organisms.     

Application of bioremediation technology in the environment contaminated with petroleum hydrocarbon

The demand for petroleum and petroleum products is increasing day by day, while oil spills and improper discharge of industrial wastes contributes to the rising contamination of the environment with petroleum hydrocarbon. Petroleum hydrocarbon spills cause various carcinogenic and neurotoxic effects, and thus effective treatment strategies are required. The various physical and chemical methods currently in use are costly and leave toxic residues in the environment. In contrast, bioremediation is a promising technology in the treatment of petroleum hydrocarbon contamination because of its high effectiveness, lower cost and environmental synergy. Here, we review the biodegradation of petroleum hydrocarbon and various factors influencing this process.     

Inhibition by fatty acids of the biodegradation of petroleum

The extent of biodegradation of petroleum by two marine bacterial isolates was found to increase when the organisms were grown in dialysis culture. This suggests that inhibitory products are formed during growth on petroleum. Fatty acids were produced by both organisms and were present in the dialyzate (dialyzable material). Fatty acids and crude oil were found to have a synergistic toxic effect. Short-chain acids were more toxic than longer-chain ones.     

Characterization of biodegradation intermediates of nonionic surfactants by MALDI-MS. 2. Oxidative biodegradation profiles of uniform octylphenol polyethoxylate in 18O-labeled water

This paper reports the characterization of the biodegradation intermediates of octylphenol octaethoxylate (OP(8)EO) by means of matrix-assisted laser desorption/ionization mass spectrometry (MALDI-MS). The biodegradation test study was carried out in a pure culture (Pseudomonas putida S-5) under aerobic conditions using OP(8)EO as the sole carbon source and (18)O-labeled water as an incubation medium. In the MALDI-MS spectra of biodegraded samples, a series of OP(n)EO molecules with n = 2-8 EO units and their corresponding carboxylic acid products (OP(n)EC) were observed. The use of purified OP(8)EO enabled one to distinguish the shortened OPEO molecules as biodegradation intermediates. Furthermore, the formation of OP(8)EC (the oxidized product of OP(8)EO) supported the notion that terminal oxidation is a step in the biodegradation process. When biodegradation study was carried out in (18)O-labeled water, incorporation of (18)O atoms into the carboxyl group was observed for OPEC, while no incorporation was observed for the shortened OPEO products. These results could provide some rationale to the biodegradation mechanism of alkylphenol polyethoxylates.     

生物处理石油污染的研究概况

自20世纪80年代以来,人们对微生物降解石油进行了深入的研究,并逐步将这一技术应用于实际环境的处理中。处理环境中菌群的数量及组成、生物降解的有效性、实际环境中出现的影响因子对降解的影响、石油的毒性和极限环境下的生物降解石油等,许多学者都进行了广泛的研究。此外,一些研究者也尝试着通过构建生物降解模型,可以使微生物降解石油的研究能从经验上升到理论。     

Effect of shale kerogen oxidation products on biodegradation of petroleum and petroleum products in soil and water

Effect of oxidation products of shale kerogen (high-molecular-weight acids with 6-22 carbon atoms) on biodegradation of oil and oil products in soil and water was studied. High-molecular-weight acids (HMWA) not only affected the layer of oil and/or oil products and dispersed it into small particles, but also stimulated growth of Rhodococcus erythropolis VKM AS-1339D, degraders of oil and oil products. Addition of 0.001-0.003% HMWA to a medium to be purified from oil products increased the extent of bacterial biodegradation by a factor of 1.1-5.0.     

Plant Biotests for Soil and Water Contaminated with Oil and Oil Products

Reactions of higher plants (mustard, oat, rye, lettuce, dill and barley) and microalgae (Euglena gracilis) to the contamination of soil and water with oil and oil products was studied. The germination of seeds was analyzed. The length of sprouts, dry biomass and length of plant roots, as well as the optical density of microalgal broth culture were determined. Negative effects of soil and water contamination with oil and oil products on plant and microalgal parameters examined was shown. After biological destruction of contaminants by an association of destructor strains (Acinetobactersp., Mycobacterium flavescens andRhodoccocussp.), the toxicity of contaminated mediums decreased. The data suggest that the integral toxicity of soil and water contaminated with oil and oil products and toxicity changes during biodestruction of these pollutants can be analyzed by using plant test organisms.     

Phytoremediation of petroleum hydrocarbons by using a freshwater fern species Azolla filiculoides Lam

In this study, the phytoremediation capacity of Azolla filiculoides Lam. for the water resources contaminated with petroleum hydrocarbons was investigated. The plants were grown in nitrogen-free Hoagland nutrient solution containing 0.005%, 0.01%, 0.05%, 0.1%, 0.2%, 0.3%, 0.4%, and 0.5% crude oil under greenhouse conditions for 15 days. Although the growth rate of the plants were not negatively influenced by the presence of crude oil in the media for the concentration of 0.005% and 0.01% v/v, a gradual impeding effect of crude oil in the growth media has been observed at concentrations 0.05–0.1%. More than 0.1% crude oil in the growth medium ostensibly retarded the growth. For example, 0.2% oil in the media reduced growth approximately 50% relative to the control, and the presence of crude oil at concentrations 0.3% or more were lethal. The data about the percentage of plant growth, fresh weight increase and root growth clearly indicated that the tolerance level of A. filiculoides plants to crude oil ranges between 0.1% and 0.2%. In comparison to control samples, the biodegradation rate of total aliphatic and aromatic (phenathrene) hydrocarbons at 0.05–0.2% oil concentrations, was 94–73% and 81–77%, respectively. On the other hand, in case of further increases in oil concentration in media, i.e.; 0.3–0.5%, the biodegradation rate was still higher in the experimental samples, respectively 71–63% and 75–71%. The high biodegradation rates of petroleum hydrocarbons in the experimental samples suggested that A. filiculoides plants could be a promising candidate to be used for the phytoremediation of low crude oil contaminated precious freshwater resources.     

Allocation of energy use in petroleum refineries to petroleum products

Aim, Scope, and Background  Studies to evaluate the energy and emission impacts of vehicle/fuel systems have to address allocation of the energy use and emissions associated with petroleum refineries to various petroleum products because refineries produce multiple products. The allocation is needed in evaluating energy and emission effects of individual transportation fuels. Allocation methods used so far for petroleum-based fuels (e.g., gasoline, diesel, and liquefied petroleum gas [LPG]) are based primarily on mass, energy content, or market value shares of individual fuels from a given refinery. The aggregate approach at the refinery level is unable to account for the energy use and emission differences associated with producing individual fuels at the next sub-level: individual refining processes within a refinery. The approach ignores the fact that different refinery products go through different processes within a refinery. Allocation at the subprocess level (i.e., the refining process level) instead of at the aggregate process level (i.e., the refinery level) is advocated by the International Standard Organization. In this study, we seek a means of allocating total refinery energy use among various refinery products at the level of individual refinery processes. Main Features  We present a petroleum refinery-process-based approach to allocating energy use in a petroleum refinery to petroleum refinery products according to mass, energy content, and market value share of final and intermediate petroleum products as they flow through refining processes within a refinery. The approach is based on energy and mass balance among refining processes within a petroleum refinery. By using published energy and mass balance data for a simplified U.S. refinery, we developed a methodology and used it to allocate total energy use within a refinery to various petroleum products. The approach accounts for energy use during individual refining processes by tracking product stream mass and energy use within a refinery. The energy use associated with an individual refining process is then distributed to product streams by using the mass, energy content, or market value share of each product stream as the weighting factors. Results  The results from this study reveal that product-specific energy use based on the refinery process-level allocation differs considerably from that based on the refinery-level allocation. We calculated well-to-pump total energy use and greenhouse gas (GHG) emissions for gasoline, diesel, LPG, and naphtha with the refinery process-based allocation approach. For gasoline, the efficiency estimated from the refinery-level allocation underestimates gasoline energy use, relative to the process-level based gasoline efficiency. For diesel fuel, the well-to-pump energy use for the process-level allocations with the mass- and energy-content-based weighting factors is smaller than that predicted with the refinery-level allocations. However, the process-level allocation with the market-value-based weighting factors has results very close to those obtained by using the refinery-level allocations. For LPG, the refinery-level allocation significantly overestimates LPG energy use. For naphtha, the refinery-level allocation overestimates naphtha energy use. The GHG emission patterns for each of the fuels are similar to those of energy use. Conclusions  We presented a refining-process-level-based method that can be used to allocate energy use of individual refining processes to refinery products. The process-level-based method captures process-dependent characteristics of fuel production within a petroleum refinery. The method starts with the mass and energy flow chart of a refinery, tracks energy use by individual refining processes, and distributes energy use of a given refining process to products from the process. In allocating energy use to refinery products, the allocation method could rely on product mass, product energy contents, or product market values as weighting factors. While the mass- and energy-content-based allocation methods provide an engineering perspective of energy allocation within a refinery, the market-value-based allocation method provides an economic perspective. The results from this study show that energy allocations at the aggregate refinery level and at the refining process level could make a difference in evaluating the energy use and emissions associated with individual petroleum products. Furthermore, for the refining-process-level allocation method, use of mass — energy content- or market value share-based weighting factors could lead to different results for diesel fuels, LPG, and naphtha. We suggest that, when possible, energy use allocations should be made at the lowest subprocess level — a confirmation of the recommendation by the International Standard Organization for life cycle analyses. Outlook  The allocation of energy use in petroleum refineries at the refining process level in this study follows the recommendation of ISO 14041 that allocations should be accomplished at the subprocess level when possible. We developed a method in this study that can be readily adapted for refineries in which process-level energy and mass balance data are available. The process-level allocation helps reveal some additional energy and emission burdens associated with certain refinery products that are otherwise overlooked with the refinery-level allocation. When possible, process-level allocation should be used in life-cycle analyses.     

The potential for hydrocarbon biodegradation and production of extracellular polymeric substances by aerobic bacteria isolated from a Brazilian petroleum reservoir

Extracellular polymeric substances (EPS) can contribute to the cellular degradation of hydrocarbons and have a huge potential for application in biotechnological processes, such as bioremediation and microbial enhanced oil recovery (MEOR). Four bacterial strains from a Brazilian petroleum reservoir were investigated for EPS production, emulsification ability and biodegradation activity when hydrocarbons were supplied as substrates for microbial growth. Two strains of Bacillus species had the highest EPS production when phenanthrene and n-octadecane were offered as carbon sources, either individually or in a mixture. While Pseudomonas sp. and Dietzia sp., the other two evaluated strains, had the highest hydrocarbon biodegradation indices, EPS production was not detected. Low EPS production may not necessarily be indicative of an absence of emulsifier activity, as indicated by the results of a surface tension reduction assay and emulsification indices for the strain of Dietzia sp. The combined results gathered in this work suggest that a microbial consortium consisting of bacteria with interdependent metabolisms could thrive in petroleum reservoirs, thus overcoming the limitations imposed on each individual species by the harsh conditions found in such environments.     

Potential of vetiver (vetiveria zizanioides (L.) Nash) for phytoremediation of petroleum hydrocarbon-contaminated soils in Venezuela

Venezuela is one of the largest oil producers in the world. For the rehabilitation of oil-contaminated sites, phytoremediation represents a promising technology whereby plants are used to enhance biodegradation processes in soil. A greenhouse study was conducted to determine the tolerance of vetiver (Vetiveria zizanioides (L.) Nash) to a Venezuelan heavy crude oil in soil. Additionally, the plant's potential for stimulating the biodegradation processes of petroleum hydrocarbons was tested under the application of two fertilizer levels. In the presence of contaminants, biomass and plant height were significantly reduced. As for fertilization, the lower fertilizer level led to higher biomass production. The specific root surface area was reduced under the effects of petroleum. However, vetiver was found to tolerate crude-oil contamination in a concentration of 5% (w/w). Concerning total oil and grease content in soil, no significant decrease under the influence of vetiver was detected when compared to the unplanted control. Thus, there was no evidence of vetiver enhancing the biodegradation of crude oil in soil under the conditions of this trial. However, uses of vetiver grass in relation to petroleum-contaminated soils are promising for amelioration of slightly polluted sites, to allow other species to get established and for erosion control.     

Investigation of the sublethal effects of some petroleum refinery effluents

In Canada, environmental regulations for protection of the biota from the adverse effects of effluents from petroleum refineries have tended to focus on acute toxicity. There is concern those effluents may have other subtle, but still deleterious, long-term effects on aquatic ecosystems. We have used a battery of toxicity tests to assess the acute toxicity, genotoxicity, and chronic toxicity of effluent samples from two Ontario refineries. The test organisms included representatives of the bacterial, algal, plant, cladoceran, and fish communities. The results of our preliminary study indicate that the effluent samples had little acute toxicity to the test organisms. There were indications of some sublethal toxicity to Ceriodaphnia dubia, Panagrellus redivivus, and Pimephales promelas. One of the effluents inhibited the growth of Selanastrum capricornutum (IC₅₀ of 59.9%) and Lemna gibba (IC₂₅ of 73.3%) and also caused a 15 percent reduction in the germination of Lactuca sativa seeds. The SOS-Chromotest, a commercially available test that measures the activity of a bacterial DNA repair system, detected genotoxic effects in a single effluent that had been concentrated ten fold. There was no apparent relationship between several chemical parameters and the observed sublethal effects. Further research is needed to establish whether or not the observed toxic effects are typical of effluents from Ontario refineries.     

Impact of surrogate selection on risk assessment for total petroleum hydrocarbons

Environmental contamination involving total petroleum hydrocarbons (TPH) is being investigated and remediated at underground storage tanks, tank farms, pipelines, and refineries across the country. Human health and environmental risk play a significant role in decision making at these sites. However, risk assessment for sites contaminated with petroleum products typically is complicated by inadequate information about the composition of TPH present at the site and the physical and chemical properties and toxicity of the components. To address these data gaps, risk assessors can select surrogate compounds to represent the movement of TPH in the environment at the site and toxicity of TPH present at the site. This article illustrates the potential impact of choice of surrogates on risk estimates, which in turn affect remediation costs.     

Controlled Release Fertilizer Increased Phytoremediation of Petroleum-Contaminated Sandy Soil

A greenhouse experiment was conducted to determine the effect of the application of controlled release fertilizer [(CRF) 0, 4, 6, or 8 kg m^–3] on Lolium multiflorum Lam. survival and potential biodegradation of petroleum hydrocarbons (0, 3000, 6000, or 15000 mg kg^–1) in sandy soil. Plant adaptation, growth, photosynthesis, total chlorophyll, and proline content as well as rhizosphere microbial population (culturable heterotrophic fungal and bacterial populations) and total petroleum hydrocarbon (TPH)-degradation were determined. Petroleum induced-toxicity resulted in reduced plant growth, photosynthesis, and nutrient status. Plant adaptation, growth, photosynthesis, and chlorophyll content were enhanced by the application of CRF in contaminated soil. Proline content showed limited use as a physiological indicator of petroleum induced-stress in plants. Bacterial and filamentous fungi populations were stimulated by the petroleum concentrations. Bacterial populations were stimulated by CRF application. At low petroleum contamination, CRF did not enhance TPH-degradation. However, petroleum degradation in the rhizosphere was enhanced by the application of medium rates of CRF, especially when plants were exposed to intermediate and high petroleum contamination. Application of CRF allowed plants to overcome the growth impairment induced by the presence of petroleum hydrocarbons in soils.     

陕北地区石油污染土壤中不动杆菌属的筛选、鉴定及降解性能

从陕北地区石油污染土壤中分离鉴定得到两株不动杆菌属(Acinetobacter sp.)的高效石油降解菌A.sp 1和A.sp 2,分别从盐浓度、pH值、氮源、磷源和接种量等因素进行研究以确定其最佳石油降解条件,并进一步通过GC-MS(Gas ChromatographyMass Spectrometer)方法分析其在最佳条件下对原油组分的不同降解性能。结果显示:A.sp 1在盐浓度为1%、pH值为6—7、磷源为KH2PO4和K2HPO4、氮源为尿素和接种量为4%的条件下,最高降解率可达到60%。A.sp 2在盐浓度为1%、pH值为7—9、磷源为KH2PO4和K2HPO4、氮源为硝酸铵和接种量为8%的条件下,最高降解率可达到67%。GC-MS分析结果表明,菌株A.sp 1对石油烃类C21—C25有明显的降解效果,菌株A.sp 2对石油烃类C20—C30的降解效果较好。     

Application of the OECD 301F respirometric test for the biodegradability assessment of various potential endocrine disrupting chemicals

The biodegradability of several potential endocrine disrupting compounds, namely 4-n-nonylphenol (4-n-NP), nonylphenol monoethoxylate (NP1EO), nonylphenol diethoxylate (NP2EO), bisphenol A (BPA), triclosan (TCS), di-(2-ethylhexyl)-phthalate (DEHP), perfluorooctanoate (PFOA) and perfluorononanoate (PFNA) was evaluated in this study, using OECD method 301F (manometric respirometry test) and activated sludge as inoculum. According to the results, 4-n-NP and BPA meet the strict definition of ready biodegradability and they are not expected to be persistent during the activated sludge process. Partial biodegradation was observed for DEHP (58.7+/-5.7%, n=3), TCS (52.1+/-8.5%, n=3) and NP1EO (25.9+/-8.1%, n=3), indicating their possible biodegradation in wastewater treatment systems, while no biodegradation was observed for NP2EO, PFOA and PFNA. Experiments in the co-presence of a readily biodegradable compound showed the absence of co-metabolic phenomena during 4-n-NP, BPA and TCS biodegradation. Using first order kinetics to describe biodegradation of the target compounds, half-lives of 4.3+/-0.6, 1.3+/-0.2, 1.8+/-0.5, 6.9+/-2.6 days were calculated for 4-n-NP, BPA, TCS and DEHP, respectively. Toxicity tests using marine bacterium Vibrio fischeri showed that biodegradation of 4-n-NP, NP1EO, BPA and TCS is a simultaneous detoxification process, while possible abiotic or biotic transformations of NP2EO, DEHP, PFOA and PFNA during respirometric test resulted to significant increase of their toxicities.     

Rhodococcus sp. F92 immobilized on polyurethane foam shows ability to degrade various petroleum products

This work reports on the immobilization and performance of a hydrocarbon-degrading microorganism on polyurethane foam (PUF) in the bioremediation of petroleum hydrocarbons. The ability of four different microorganisms to immobilize on PUF and to degrade various petroleum products (Arabian light crude (ALC), Al-Shaheen crude (ASC), diesel and oil slops) was assessed by measuring the n-alkane fraction remaining in the petroleum products over time. A Rhodococcus sp. (designated as F92) had the highest number of immobilized viable cells (10(9) cells per cm3 PUF) and a maximum attachment efficiency of 90% on PUF of a density of 14 kg/m3. Scanning electron microscopy showed the presence of extracellular structures that could play an important role in the immobilization of F92 on PUF. Analysis by GC-MS revealed that both free and immobilized F92 cells were able to degrade approximately 90% of the total n-alkanes in the petroleum products tested within 1 week at 30 degrees C. Rhodococcus sp. F92 was efficiently immobilized onto PUF and the immobilized cells were able to degrade a variety of petroleum products such as ALC, ASC, diesel and oil slops. The results suggest the potential of using PUF-immobilized Rhodococcus sp. F92 to bioremediate petroleum hydrocarbons in an open marine environment.     

微生物强化修复石油污染土壤的研究进展

微生物修复被认为是去除石油污染物和修复石油污染土壤的一种经济、高效且无二次污染的绿色清洁技术。受土壤环境条件和石油污染物性质等因素制约,土壤中土著石油降解微生物常存在数量不足、活性偏低、生长缓慢等问题,导致修复效果不佳、修复周期偏长。微生物强化修复技术可有效提高微生物降解效能,通过投加具有降解效能的功能菌株或菌剂、营养物质、表面活性剂、生长基质及固定化微生物等手段,可改善提升土著微生物对石油污染土壤的修复效果。文中梳理了已报道的石油降解微生物的种类,总结了微生物修复石油污染土壤的主要影响因素,阐述了微生物强化修复石油土壤的多种有效策略,提出了微生物强化修复石油污染的未来发展方向。     

Biodegradation of wood in crude oil-polluted soil

A field investigation (April–November) in Nigeria showed that biodegradation of obeche (Triplochiton scleroxylon) wood blocks was initially retarded in crude oil-contaminated soil but later became enhanced as indicated by loss of compression resistance. Further indication of this pattern was the detection of soft-rot cavities and basidiomycete fungi after 2–3 months exposure when compared to control blocks in uncontaminated soil. Laboratory tests with Pleurotus sp., Trametes sp., Gloeophyllum sp. (basidiomycetes) and Chaetomium sp. (soft-rot fungus) confirmed that degradation of crude oil-coated obeche blocks was markedly retarded without the presence of hydrocarbon-degrading bacteria. The filtrate of hydrocarbon-degrading Pseudomonas sp. grown in mineral salt/crude oil medium for 3–4 weeks supported growth of the test fungi better than in carboxymethyl cellulose medium but less than in potato dextrose broth. Similarly, wood blocks immersed in the filtrate became significantly more susceptible to fungal degradation. Pseudomonas sp. from stationary phase growth in crude oil medium depleted residual sugar in basidiomycete-degraded sawdust with a concomitant marked increase in its population. It may be concluded that readily metabolizable products of crude oil degradation by soil organisms and the removal of residual sugar which may have prevented catabolite repression of cellulases, culminated in increased attack on the wood by soil-borne wood-decomposing organisms.