Microbial Factors Rather Than Bioavailability Limit the Rate and Extent of PAH Biodegradation in Aged Crude Oil Contaminated Model Soils

The rate and extent of polynuclear aromatic hydrocarbons (PAH) biodegradation in a set of aged model soils that had been contaminated with crude oil at the high concentrations (i.e.,>20,000?mg/kg) normally found in the environment were measured in 90-week slurry bioremediation experiments. Soil properties such as organic matter content, mineral type, particle diameter, surface area, and porosity did not significantly influence the PAH biodegradation kinetics among the 10 different model soils. A comparison of aged and freshly spiked soils indicates that aging affects the biodegradation rate and extent only for higher-molecular-weight PAHs, while the effects of aging are insignificant for 4-ring PAHs and total PAHs. In all model soils with the exception of kaolinite clay, the rate of abiotic desorption was faster than the rate of biodegradation during the initial phase of bioremediation treatment, indicating that PAH biodegradation was limited by microbial factors. Similarly, any of the higher-molecular-weight PAHs that were still present after 90 weeks of treatment were released rapidly during abiotic desorption tests, which demonstrates that bioavailability limitations were not responsible for the recalcitrance of these hydrocarbons. Indeed, an analysis of microbial counts indicates that a severe reduction in hydrocarbon degrader populations may be responsible for the observed incomplete PAH biodegradation. Therefore, it can be concluded that the recalcitrance of PAHs during bioremediation is not necessarily due to bioavailability limitations and that these residual contaminants therefore might pose a greater risk to environmental receptors than previously thought.     

Biodegradation of a PAH Mixture by Native Subsurface Microbiota

Laboratory microcosm studies were conducted to estimate biodegradation rates for a mixture of five polycyclic aromatic hydrocarbon compounds (PAHs). Static microcosms were assembled using soil samples from two locations collected at a No. 2 fuel oil-contaminated site in the Atlantic Coastal Plain of Virginia. In microcosms from one location, five PAHs (acenaphthene, fluorene, phenanthrene, pyrene, and benzo(b)fluoranthene) biodegraded at net first-order rates of 1.08, 1.45, 1.13, 1.11, and 1.12 yr^?1, respectively. No observable lag period was noted and degradation in live microcosms ceased with the depletion of oxygen and sulfate after 125 days. In microcosms from a second location, net first-order biodegradation rates after an approximately 2-month lag period were 2.41, 3.28, and 2.98 yr^?1 for fluorene, phenanthrene, and pyrene, respectively. Acenaphthene and benzo(b)fluoranthene mass loss rates in the live microcosms were not statistically different from mass loss rates in control microcosms. Stoichiometric mass balance calculations indicate that the dominant PAH mass loss mechanism was aerobic biodegradation, while abiotic losses (attributed to micropore diffusion and oxidative coupling) ranged from 15 to 33% and biotic losses from sulfate-reduction accounted for 7 to 10% of PAH mass loss. Stoichiometric equations that include biomass yield are presented for PAH oxidation under aerobic and sulfate-reducing conditions.     

Surfactant-Enhanced Biodegradation of a PAH in Soil Slurry Reactors

This study focuses on finding operational regimes for surfactant-enhanced biodegradation. Biodegradation of phenanthrene as a model poly cyclic aromatic hydrocarbon (PAH) was studied in soil slurry reactors in the presence and absence of a Triton N-101 surfactant solution. Results showed that the presence of surfactant slowed the initial biodegradation rate of phenanthrene, but increased the total mass of phenanthrene degraded over a four day period by 30%. A mathematical model was developed which simulates the biodegradation of low solubility hydrocarbons in the presence of soils and surfactants by accounting for the hydrocarbon bioavailability in different phases of the system. The model was able to simulate the experimental results using parameters and rate coefficients that were obtained through independent experiments.

The model was used to investigate the effect of different operating conditions on the overall biodegradation of phenanthrene. Simulation results showed that there is a system-specific optimum surfactant concentration range, beyond which bioremediation is hindered. The results also indicate that for a given system, the optimal surfactant concentration can be determined from simple sorption and solubility equilibrium experiments. Finally, a metric is presented for determining the potential effectiveness of surfactant-enhanced bioremediation based on the Monod and bioavailability parameters for a given system.     

Biodegradation of Hopane Prevents Use as Conservative Biomarker During Bioremediation of PAHs in Petroleum Contaminated Soils

The pentacyclic triterpane C₃₀ 17α (H), 21β (H)-hopane, a biomarker commonly used in hydrocarbon bioremediation laboratory experiments and field studies, was found to be completely removed without the formation of the demethylated intermediate nor-hopane in a crude oil-contaminated soil undergoing slurry biotreatment, while PAHs such as benzo(e)pyrene were recalcitrant. The partial or complete biodegradation of hopane has also been previously reported in a few bioremediation studies and has been explored by petroleum geochemists in an effort to characterize crude oil deposits. It is currently not clear what conditions induce hopane biodegradation or biotransformation, although the use of microbial enrichment cultures appears to speed up the process. Considering that hopane is not necessarily conserved during a bioremediation study, the uncritical normalization of hydrocarbon concentrations using this biomarker can lead to incorrect estimates of biodegradation rates and extents. If hopane is found to be unstable in a particular case, other potential biomarkers such as pentahopane, oleanane, or vanadium may be used instead.     

Differences in Sediment Organic Matter Composition and PAH Weathering between Non-Vegetated and Recently Vegetated Fuel Oiled Sediments

We examined polycyclic aromatic hydrocarbon (PAH) attenuation in contaminated field sediments after only 2 years of plant growth. We collected sediments from vegetated and non-vegetated areas at the Indiana Harbor Canal (IHC), an industrialized area with historic petroleum contamination of soils and sediments. PAH concentrations, PAH weathering indices, and organic matter composition in sediments colonized by Phragmites, cattails, or willow trees were compared to the same indices for non-vegetated sediments. We hypothesized that bulk sediment and humin fractions with measurable increases in plant organic matter content would show measurable changes to PAH attenuation as indicated by more weathered PAH diagnostic ratios or reduced PAH concentrations. Carbon-normalized PAH concentrations were lower in vegetated bulk sediments but higher in vegetated humin fractions relative to non-vegetated sediment fractions. Total organic carbon content was not indicative of more weathered N₃/P₂ ratios or reduced PAH concentrations in vegetated sediment fractions. More weathered N₃/P₂ ratios were observed with increased modern carbon (plant carbon) content of vegetated sediment fractions. Phragmites sediments contained more modern carbon (plant carbon) and more weathered PAH ratios [C₃-naphthalenes and C₂-phenanthrenes (N₃/P₂)] than willow, cattail, and non-vegetated sediments.     

Effect of Exposure Time,Contamination Level,and Type of PAH Compound on Biodegradation Capacity of Mangrove Sediment

Mangrove sediment had high natural attenuation potential with more than 50% of total PAHs being removed within 15 days. The efficiency in degrading PAHs varied with the declining order of phenanthrene (Phe), fluoranthene (Fla), and pyrene (Pyr). The Most Probable Number (MPN) of PAH-degrading bacteria in the PAH-contaminated slurries was 2 to 4 orders of magnitude higher than that in the non-contaminated mangrove slurries. The biodegradation ability of the indigenous microbial community in mangrove sediment slurry was significantly increased after exposure to polycyclic aromatic hydrocarbons. Such enhancement effect was dependent on the level and time of exposure, as well as the types of PAH compounds. The lowest contamination level of 3 mg kg^?1 was effective in promoting the degradation of Phe and Fla after seven days, but the enhancement effect for Pyr degradation was only found in the slurries exposed to contamination levels of 9 mg kg^?1 for 30 days, suggesting a threshold concentration of PAHs to stimulate growth and activity of pyrene-degrading bacteria. The contamination level higher than the threshold concentration did not lead to more degradation. The present study provides insights into the natural attenuation of PAH-contaminated mangrove sediments.     

The effect of inorganic and organic supplements on the microbial degradation of phenanthrene and pyrene in soils

The effects of several bioremediation stimulants, including potentialmetabolism pathway inducers, inorganic/organic nutrients, and surfactants onthe metabolism of phenanthrene and pyrene, as well as the populationdynamics of PAH degrading microorganisms was examined in five soils withdiffering background PAH concentrations, exposure histories and physicalproperties. Most of the supplements either had no significant effect ordecreased the mineralization of [¹⁴C]-phenanthrene and[¹⁴C]-pyrene in soil slurry microcosms. The effect of aparticular supplement, however, was often not uniform within or acrosssoils. Decreased mineralization of [¹⁴C]-phenanthrene and[¹⁴C]-pyrene was usually due to either preferential use of thesupplement as carbon source and/or stimulation of non-PAH degradingmicroorganisms. Many of the supplements increased populations ofheterotrophic microorganisms, as measured by plate counts, but did notincrease populations of phenanthrene degrading microorganisms, as measuredby the [¹⁴C]-PAH mineralization MPN analysis or cellularincorporation of [¹⁴C]-PAH. These results suggest that the PAHdegrading community at each site may be unique in their response tomaterials added in an attempt to stimulate PAH degradation. Thecharacteristics of the site, including exposure history, soil type, andtemporal variation may all influence their response.     

土壤有效硅对大豆生长发育和生理功能的影响

人工调节土壤有效硅含量及盆栽试验,研究土壤有效硅对大豆生长发育和生理功能的影响.结果表明,土壤有效硅含量在55.1～202.8mg·kg^-1范围内,随着土壤有效硅含量的提高,大豆种子萌发过程中蛋白酶和脂肪酶活性升高,淀粉酶活性无显著变化,呼吸速率加快,种子活力升高,萌发速度加快,种子萌发率无显著变化;幼苗生长过程中叶片叶绿素含量无显著变化,光合速率加快,根系活力、硝酸还原酶活力升高,蒸腾强度减弱,水分利用效率和叶含水量升高,抗旱保水能力提高.大豆幼苗含硅量与土壤有效硅含量呈线性正相关趋势(r=0.994).土壤有效硅含量大于202.8mg·kg^-1,生理功能不再显著变化,说明土壤中的硅被大豆吸收后,改善了大豆萌发种子和幼苗的生理功能,使种子萌发和幼苗生长加快.     

Bioavailability to Fish of Sediment PAH as an Indicator of the Success of In Situ Remediation Treatments at an Experimental Oil Spill

A weathered medium crude oil was applied to experimental plots of Scirpus pungens (Three-square Bulrush) in a freshwater wetland to determine the efficacy of strategies for shoreline oil spill bioremediation based on nutrient enrichment (bioremediation) and plant growth (phytoremediation). Plots were unoiled, oiled with no added nutrients, or oiled with repeated applications of phosphate and nitrate fertilizers. Following initial treatments, the experimental plots were raked to simulate the activity of wave action on oil penetration, and plants in one fertilized plot were cut repeatedly. The sediments were sampled at regular intervals for 15 months after oiling, and the loss of oil was assessed by 4-day laboratory tests of polynuclear aromatic hydrocarbon (PAH) bioaccumulation by trout, as demonstrated by increases in activity of liver cytochrome P450 (CYP1A) enzymes. Oil alone, oil mixed with sediments in the lab, and oiled sediments from treated plots all induced CYP1A activity relative to untreated controls, indicating the presence and bioavailability of PAH. Induction did not vary with nutrient treatments, but declined by 80% within 15 months of oiling, and chemical analyses indicated equivalent losses of hydrocarbons in sediment. These results demonstrate that bioavailable PAHs persisted in measurable quantities for at least 1.25 years following oiling, and that stimulation of plant growth did not affect the rate of oil disappearance. The controlling factors were likely weathering and sediment movement.     

海洋石油污染物的微生物降解与生物修复

石油是海洋环境的主要污染物 ,已经对海洋及近岸环境造成了严重的危害。微生物降解是海洋石油污染去除的主要途径。海洋石油污染物的微生物降解受石油组分与理化性质、环境条件以及微生物群落组成等多方面因素的制约 ,N和P营养的缺乏是海洋石油污染物生物降解的主要限制因子。在生物降解研究基础上发展起来的生物修复技术在海洋石油污染治理中发展潜力巨大 ,并且取得了一系列成果。介绍了海洋中石油污染物的来源、转化过程、降解机理、影响生物降解因素及生物修复技术等方面内容 ,强调了生物修复技术在治理海洋石油污染环境中的优势和重要性 ,指出目前生物修复技术存在的问题。     

二噁(口英)的生物降解及其机理

利用微生物降解二口恶口英是一种具有广阔前景的治理二口恶口英污染的方法。近年来利用微生物降解二口恶口英已经引起众多研究者的重视,国外在这方面的研究已取得了一定成果。介绍了能降解二口恶口英的微生物种类及其所能降解的二口恶口英类型,对近年来提出的微生物降解(包括氧化降解和还原降解)的可能途径,中间产物及其降解效果进行了综述。并对土壤在被污染后如何进行生物修复等方面进行了探讨。     

Improving Mathematical Model in Biodegradation of PAHs Contaminated Soil Using Gram-Positive Bacteria

In published literature there are limited studies on the estimation of kinetic parameters of polycyclic aromatic hydrocarbons (PAHs) in soil. In addition, neither the kinetic studies were performed with Gram-positive bacteria nor conducted under non-indigenous condition in order to understand their removal performance. Thus, a mathematical model describing biodegradation of phenanthrene-contaminated soil by Corynebacterium urealyticum, bacterium isolated from municipal sludge, was developed in this study. The model includes three kinetic parameters that were determined using TableCurve 2D software, namely qmax (maximum substrate utilization rate per unit mass of bacteria), X (biomass concentration) and Ks (substrate concentration at one half the maximum substrate utilization rate). These parameters were evaluated and verified in five different initial phenanthrene concentrations. Highest degradation rate was determined to be 79.24 mg kg^?1 day^?1 at 500 mg kg^?1 initial phenanthrene concentrations. This high concentration shows that bacteria perform better in contaminated sand compared to liquid media. High r² values, ranging from 0.92 to 0.99, were obtained excluding 1000 mg/kg phenanthrene. The kinetic parameters, i.e., qmax and Ks, increased with the phenanthrene concentration and thus suggest that bacteria degrade at a higher degradation rate. This model successfully described the biodegradation profiles observed at different initial phenanthrene concentrations. The established model can be used to simulate the duration of phenanthrene degradation using only the value of the initial PAHs concentration.     

Dissolved Oxygen Saturation Controls PAH Biodegradation in Freshwater Estuary Sediments

Polycyclic aromatic hydrocarbons (PAHs) are common contaminants in terrestrial and aquatic environments and can represent a significant constituent of the carbon pool in coastal sediments. We report here the results of an 18-month seasonal study of PAH biodegradation and heterotrophic bacterial production and their controlling biogeochemical factors from 186 sediment samples taken in a tidally influenced freshwater estuary. For each sampling event, measurements were averaged from 25–45 stations covering ∼250 km². There was a clear relationship between bacterial production and ambient temperature, but none between production and bottom water dissolved oxygen (DO) % saturation or PAH concentrations. In contrast with other studies, we found no effect of temperature on the biodegradation of naphthalene, phenanthrene, or fluoranthene. PAH mineralization correlated with bottom water DO saturation above 70% (r² > 0.99). These results suggest that the proportional utilization of PAH carbon to natural organic carbon is as much as three orders of magnitude higher during cooler months, when water temperatures are lower and DO % saturation is higher. Infusion of cooler, well-oxygenated water to the water column overlying contaminated sediments during the summer months may stimulate PAH metabolism preferentially over non-PAH organic matter.     

Biodegradation of Pyrene in Marine Beach Sediments

The potential of chitosan (0.1% dry weight equivalent) as a bioremediation additive for removal of the recalcitrant polycyclic aromatic hydrocarbon (PAH) pyrene in marine beach sediments was investigated using an open irrigation system over a 63-day period. Osmocote, a slow release fertilizer, was used as the key nutrient supplement at a concentration of 1% in sediment (dry weight equivalent). Osmocote significantly (p < .05) enhanced nutrient levels, and the metabolic activity of the indigenous microbial biomass. Both additives were comparable in stimulating pyrene biodegradation rates; with chitosan (0.062 day^?1) being slightly more effective as an amendment than Osmocote (0.051 day^?1). Loss of pyrene in a control sediment (i.e., pyrene, without additives) was 66.6% over a 63-day period. The concurrent application of additives yielded the greatest biodegradation rates (0.072day^?1), resulting in a 98.2% loss of pyrene over 63 days. The treatment of oil contaminated beach sediments with both osmocote (1%) and chitosan (0.1%) is therefore recommended as an effective treatment for the intrinsic biodegradation of recalcitrant PAHs in oil-contaminated beach sediments.     

Batch Leaching Tests: Colloid Release and PAH Leachability

The Toxicity Characteristic Leaching Procedure (TCLP) was developed by the U.S. Environmental Protection Agency to assess leaching potential of contaminants from waste, and to provide a test to classify hazardous waste. It is a batch leaching test where a waste (such as contaminated soil) and an extraction fluid are agitated for a predetermined time. Since TCLP employs an aggressive mixing technique, it is possible that hydrophobic contaminant-laden colloidal fractions may appear as “dissolved” constituents. In this study, TCLP was employed to determine the leachability of PAH contamination from a coal tar contaminated site. Generated colloids and the apparent aqueous concentrations of naphthalene and phenanthrene were measured at various mixing times in the extraction fluid.

A mathematical model was developed that predicted the apparent aqueous contaminant concentration in the filtrate. This model accounted for the presence of colloids in the filtrate, and quantified contaminant desorption from colloids. The fraction of colloid-bound contaminant was predicted to be negligible for naphthalene. However, phenanthrene was predicted to have a significant fraction of the total contaminant in the colloidal phase, while naphthalene was primarily dissolved. The desorption model and PAH desorption data are presented here to determine the extent of colloid-facilitated desorption during leaching tests.     

TGF-beta1 通过PI3K及ERK信号通路调节肺动脉高压过程中IGFBPs蛋白表达

目的：探讨大鼠肺动脉高压(PAH)过程中TGF-beta1对胰岛素样生长因子结合蛋白（IGFBP）表达调节是否依赖于PI3K及ERK 信号通路。方法：取健康成年SD 大鼠26 只,随机分成2 组：PAH组,腹腔注射1%的野百合碱,剂量为60 mg/kg;对照(C)组腹腔 注射生理盐水。于4 周后超声检测肺动脉平均压力,取肺组织做HE 染色,应用NIS-Element 系统测量中膜厚度。原代培养肺动脉 平滑肌（PASMC）细胞,分别加入TGF-beta1 及TGF-beta1 中和抗体后,Western-blot 检测IGFBP3,IGFBP5,Smad2/Smad3 表达。加入 ERK特异性抑制剂PD98059 或PI3K 抑制剂LY294002,检测IGFBP3,IGFBP5 表达。结果：野百合碱处理4 周后,肺动脉高压组 的平均肺动脉压力及右室/(左室+室间隔)比值显著高于对照组。TGF-茁1 可显著升高IGFBP3,IGFBP5 及p-Smad3 的表达（P<0. 05）,而抑制TGF-beta1 则可显著降低三种蛋白的表达（P<0.05）。加入LY294002 抑制PI3K ERK 后,IGFBP3 和p-Smad2 两种蛋白的 表达量显著下调（P<0.05）。加入PD98059 抑制ERK 后可显著降低IGFBP3 及IGFBP5 的表达水平（P<0.05）。结论：PAH 中 TGF-茁1 升高可通过活化Smad2/Smad3 上调IGFBP3和IGFBP5 的表达。TGF-beta1 促进IGFBP3,IGFBP5表达的作用依赖于PI3K 及ERK 信号通路。     

Studies on Biodegradation of Kerosene in Soil under Different Bioremediation Strategies

The effectiveness of bioremediation is often a function of the microbial population and how they can be enriched and maintained in an environment. Strategies for inexpensive in situ bioremediation of soil contaminated with petroleum hydrocarbons include stimulation of the indigenous microorganisms by introduction of nutrients (biostimulation) and/or through inoculation of an enriched mixed microbial culture into soil (bioaugmentation). To demonstrate the potential use of bioremediation in soil contaminated with kerosene, a laboratory study with the objective of evaluating and comparing the effects of bioattenuation, biostimulation, bioaugmentation, and combined biostimulation and bioaugmentation was performed. The present study dealt with the biodegradation of kerosene in soil under different bioremediation treatment strategies: bioattenuation, biostimulation, bioaugmentation, and combined biostimulation and bioaugmentation, respectively. Each treatment strategy contained 10% (w/w) kerosene in soil as a sole source of carbon and energy. After 5 weeks of remediation, the results revealed that bioattenuation, bioaugmentation, biostimulation, and combined biostimulation and bioaugmentation exhibited 44.1%, 67.8%, 83.1%, and 87.3% kerosene degradation, respectively. Also, the total hydrocarbon-degrading bacteria (THDB) count in all the treatments increased with time up till the second week after which it decreased. The highest bacterial growth was observed for combined biostimulation and bioaugmentation treatment strategy. A first-order kinetic model equation was fitted to the biodegradation data to further evaluate the rate of biodegradation and the results showed that the specific degradation rate constant (k) value was comparatively higher for combined biostimulation and bioaugmentation treatment strategy than the values for other treatments. Therefore, value of the kinetic parameter showed that the degree of effectiveness of these bioremediation strategies in the clean up of soil contaminated with kerosene is in the following order: bioattenuation < bioaugmentation < biostimulation < combined biostimulation and bioaugmentation. Conclusively, the present work has defined combined biostimulation and bioaugmentation treatment strategy requirements for kerosene oil degradation and thus opened an avenue for its remediation from contaminated soil.     

NATIVE MICHIGAN PLANTS STIMULATE SOIL MICROBIAL SPECIES CHANGES AND PAH REMEDIATION AT A LEGACY STEEL MILL

A 1.3-acre phytoremediation site was constructed to mitigate polyaromatic hydrocarbon (PAH) contamination from a former steel mill in Michigan. Soil was amended with 10% (v/v) compost and 5% (v/v) poultry litter. The site was divided into twelve 11.89 m X 27.13 m plots, planted with approximately 35,000 native Michigan perennials, and soils sampled for three seasons. Soil microbial density generally increased in subplots of Eupatorium perfoliatum (boneset), Aster novae-angliae (New England aster), Andropogon gerardii (big bluestem), and Scirpus atrovirens (green bulrush) versus unplanted subplots. Using enumeration assays with root exudates, PAH degrading bacteria were greatest in soils beneath plants. Initially predominant, Arthrobacter were found capable of degrading a PAH cocktail in vitro, especially upon the addition of root exudate. Growth of some Arthrobacter isolates was stimulated by root exudate. The frequency of Arthrobacter declined in planted subplots with a concurrent increase in other species, including secondary PAH degraders Bacillus and Nocardioides. In subplots supporting only weeds, an increase in Pseudomonas density and little PAH removal were observed. This study supports the notion that a dynamic interplay between the soil, bacteria, and native plant root secretions likely contributes to in situ PAH phytoremediation.     

Toxicity of phenanthrene dissolved in nonionic surfactant solutions to Pseudomonas putida P2

The fraction in which direct contact occurs between micellar-phase phenanthrene and the bacterial cell surface was estimated by measuring the toxicity of nonionic surfactant (Tween 80 and Triton X-100) solutions to the phenanthrene-degrading bacterium, Pseudomonas putida P2. Cell viability of completely dissolved phenanthrene decreased by 30% at concentrations greater than 0.3 mg L(-1), which is equal to approximately one third of its solubility. Both nonionic surfactants had no effect on cell viability up to 5 g L(-1). Cell viability increased with increasing surfactant concentration at a fixed phenanthrene concentration, due to the decreased concentration of aqueous-pseudophase phenanthrene and the reduced fraction of direct contact. The fraction of direct contact was c. 20% or more below 3 g L(-1) of Triton X-100. The fraction of direct contact for Tween 80 was estimated to be lower than Triton X-100.