Glycine-β-cyclodextrin-assisted cometabolism of phenanthrene and pyrene by Pseudomonas stutzeri DJP 1 from marine sediment

Cometabolic degradation is currently an effective and extensively way to remove high molecular weight polycyclic aromatic hydrocarbons(HMW-PAHs).Unfortunately,due to low bio-accessibility and high biotoxicity,the cometabolic degradation rate of HMW-PAHs is limited.Glycine-β-cyclodextrin(GCD)was obtained through amino modification ofβ-cyclodextrin(BCD)and added to cometabolic system of phenanthrene(PHE)and pyrene(PYR)to assist PYR biodegradation.Results show that the addition of GCD(100 mg/L)effectively improved the removal rate of PYR(20 mg/L)by 42.3%.GCD appeared to increase the bio-accessibility and reduce the biotoxicity of PHE and PYR,and then promoted the growth of Pseudomonas stutzeri DJP1 and stimulated the elevation of dehydrogenase(DHA)and catechol 12 dioxygenase(C12O)activities.The phthalate metabolic pathway was accelerated,which improved the cometabolic degradation.This study provided a new reference for the cometabolic degradation of HMW-PAHs.

Effects of Fe3+ and Ca2+ on sorption of phenanthrene by Humin in karst soil,Southwest China

Humin(HM) is the main organic matter component to af fect the migration and transformation of polycyclic aromatic hydrocarbons(PAHs) in soil. The study on infl uence of the morphology change of inorganic ions on the adsorption of PAHs in soil and its organic matter is still rare at microscopic scale. In this paper, yellow soil humin(YS-HM) and lime soil humin(LS-HM) were chosen as samples, then Fe^(3+) and Ca^(2+) were added into samples to facilitate the precipitation by changing the existing conditions of ions, and the mechanism by which inorganic precipitation changed adsorption capacity of karst soil was analyzed from the microscopic scale. The results showed that the adsorption capacity of HM reduced with the inorganic precipitation increasing. The precipitation of Ca^(2+)and Fe^(3+) both reduced the adsorption capacity of YS-HM and LS-HM by 61.71% and 71.83% on average, respectively. The results of scanning electron microscope-energy dispersive spectrometry(SEM-EDS) and pore analysis showed that the HM porosity decreased after formation of Ca^(2+) and Fe^(3+) precipitation. According to the value of Freundlich parameter n, it may be because the precipitation or colloid of Ca^(2+) and Fe^(3+) fi lled micropores and covers high-energy adsorption sites of the HM. This research provides theoretical support for studying the PAHs migration and bioavailability of Calcium-rich in karst soil.

Antioxidant phenanthrenes and lignans from Dendrobium nobile

To study the antioxidant constituents from the stems of Dendrobium nobile, and to discuss theft structure-activity relationship. Compounds were isolated from a 60% ethanolic extract by various chromatographic techniques and were identified by spectral analysis. The antioxidant activities of compounds were evaluated by DPPH free radical scavenging assay. Five phenanthrenes and four lignans were obtained from the active fractions ofD. nobile. Their structures were identified as fimbriatone （1）, confusarin （2）, flavanthrinin （3）, 2,5-dihydroxy-4,9-dimethoxyphenanthrene （4）, 3,7-dihydroxy-2,4-dimethoxyphenanthrene （5）, syringaresinol （6）, pinoresinol （7）, medioresinol （8） and lirioresinol-A （9）, respectively. Compounds 2 and 6 exhibited more potent DPPH scavenging activities than vitamin C. All the above compounds were reported from this plant for the first time, and compounds 3, 4 and 9 were reported for the first time from the genus of Dendrobiurn. For all phenanthrenes and lignans, an electron-donating methoxyl group in the ortho position to the phenolic hydroxyl group exhibits enhanced antioxidant activities.

Evaluation of genotoxicity of combined soil pollution by cadmium and phenanthrene on earthworm

The DNA-damaging effects of the combined pollution of heavy metal and organic contamination on earthworms were evaluated by single cell gel electrophoresis （SCGE） assay. Earthworms Eisenia andrei were exposed to single or combined test compounds in different doses of cadmium （Cd） 5, 10, 50 mg/kg and phenanthrene （Phe） 0.5, 2.5, 12.5 mg/kg with a treatment of 14 d. In SCGE assay, isolated coelomcytes and electrophoresis were employed to determine DNA damage degree after a 14-d treatment by test compounds. The results showed that there was a significant statistical difference between earthworms treated with Cd combined Phe with them treated alone with Cd or Phe. The Olive tail moment （OTM） of SCGE assay using earthworm coelomcytes appears to be a sensitive biomarker for evaluating exposure to genotoxic compounds. These tests also revealed that the interaction between Cd and Phe to DNA damaging effects was negative, and was strongly dependent on the concentration of pollution. This study corresponds to exploratory phase in order to reveal interaction effects of heavy metal and organic contamination on earthworms and then to set up more in-depth analysis to increase progressively the understanding of the genotoxicity mechanisms involved.

Effects of temperature and surfactants on naphthalene and phenanthrene sorption by soil

Adsorption experiments were carried out to investigate the sorption behaviors of naphthalene and phenanthrene in six different soils and to determine the effects of temperature, linear alkylbenzene sulfonate （LAS） and cetylrimethyl ammonium bromide （CTAB） on sorption. The results show that for a given sorbent phenanthrene exhibited greater nonlinear and stronger sorption than naphthalene. There was a strong negative correlation for the Koc values with organic carbon content （foc）. The increase of temperature was not favorable to sorption. Sorption decreased along with the increasing aqueous LAS concentration from 0 to 1000 mg/L. At low CTAB concentration （〈 100 mg/L）, the adsorption increased as CTAB hemimicelles formed on the soil surface. At high concentration, CTAB decreased the adsorption by occupying active hydrophobic adsorption sites and solubilization of naphthalene and phenanthrene.

Phytoremediation for phenanthrene and pyrene contaminated soils

Phytoremediation of soil contaminated with phenanthrene and pyrene was investigated using twelve plant species. Plant uptake and accumulation of these chemicals were evaluated. At the end of the experiment(45 d), the remaining respective concentrations of soil phenanthrene and pyrene in spiked vegetated soils, with initial phenanthrene of 133 3 mg/kg and pyrene of 171 5 mg/kg, were 8 71—16 4 and 44 9—65 0 mg/kg, generally 4 7%—49 4% and 7 1%—35 9% lower than their concentrations in the non vegetated soils. The loss of phenanthrene and pyrene in vegetated spiked soils were 88 2%—93 0% and 62 3%—73 8% of the added amounts of these contaminants, respectively. Although plant uptake and accumulation of these compounds were evident, and root concentrations and RCFs(root concentration factors; defined as the ratio of PAH concentrations in roots and in the soils on a dry weight basis) of these compounds significantly positively correlated to root lipid contents, plant uptake and accumulation only accounted for less than 0 01% and 0 23% of the enhanced loss of these chemicals in vegetated versus non vegetated soils. In contrast, plant promoted microbial biodegradation was the dominant mechanism of the phytoremediation for soil phenanthrene and pyrene contamination. Results from this study suggested a feasibility of the establishment of phytoremediation for soil PAH contamination.

Effects of maize root exudates and organic acids on the desorption of phenanthrene from soils

The effects of maize root exudates and low-molecular-weight-organic anions （LMWOAs） on the desorption of phenanthrene from eight artificially contaminated soils were evaluated. A significant negative correlation was observed between the amounts of phenanthrene desorbed and the soil organic carbon （SOC） contents （P 〈 0.01）, and the influences of soil pH and clay content on phenanthrene desorption were insignificant （P 〉 0.1）. Neither maize root exudates nor oxalate and citrate anions influenced desorption of phenanthrene with the addition of NaN3. A faster phenanthrene desorption occurred without the addition of NaN3 in the presence of maize root exudates than oxalate or citrate due to the enhanced degradation by root exudates. Without the addition of NAN3, oxalate or citrate at different concentrations could inhibit phenanthrene desorption to different extents and the inhibiting effect by citrate was more significant than by oxalate. This study leads to the conclusion that maize root exudates can not enhance the desorption under abiotic condition with the addition of NaN3 and can promote the desorption of phenanthrene in soils without the addition of NaN3.

Influence of nonionic surfactant on the solubilization and biodegradation of phenanthrene

Phenanthrene was solubilized in two different nonionic surfactants, Tween80 and Triton X-100. The bioavailability of phenanthrene to the bacteria isolated from the petroleum contaminated soils was studied based on the rotary flasks experiments. The results showed that the concentration of nonionic surfactants above the critical micelle concentration(CMC) can increase the solubility of phenanthrene in water and were innoxious to the phenanthrene-degrading bacteria; phenanthrene solubilized in the micelles of Tween80 was bioavailable and biodegradable. The research demonstrated the potential of surfactant-enhanced bioremediation of soils contaminated by hydrophobic organic compounds(HOCs).

Interactive effect of dissolved organic matter and phenanthrene on soil enzymatic activities

The investigation of dissolved organic matter （DOM） on urease, catalase and polyphenol oxidase activity in a phenanthrene （Phe）- contaminated soil was conducted under laboratory incubation conditions. Values of soil enzymatic activity depended mainly on incubation time. In the initial 16 days, urease activity increased, and was followed by a decrease. In the initial 8 days, catalase activity decreased and then increased. Variation of polyphenol oxidase activity was just the reverse of catalase activity. After 30 days of incubation, no pronounced difference among treatments with Phe, Phe and DOM, and control were detected in urease, catalase and polyphenol oxidase activity. Phe might inhibit urease and catalase, and stimulate polyphenol oxidase. DOM could improve inhibition of Phe in soil urease and catalase activity during the initial period of applying DOM. Nevertheless, DOM had no significant effect on polyphenol oxidase activity in the Phe contaminated soil. There was a negative correlation between catalase and polyphenol oxidase （r = -0.761＊＊＊）, and catalase and urease （r = -0.554＊＊）. Additionally, a positive correlation between polyphenol oxidase and urease was also detected （r = 0.701＊＊＊）. It is implied that the formed DOM after application of organic wastes into soils may counteract the inhibition of polycyclic aromatic hydrocarbons in soil enzyme activities.

Characterization of phenanthrene degradation by strain Polyporus sp. S133

Polyporus sp. S133, a fungus collected from contaminated soil, was used to degrade phenanthrene, a polycyclic aromatic hydrocarbon, in a mineral salt broth liquid culture. A maximal degradation rate （92%） was obtained when Polyporus sp. S133 was cultured for 30 days with agitation at 120 r/min, as compared to 44% degradation in non-agitated cultures. Furthermore, the degradation was affected by the addition of surfactants. Tween 80 was the most suitable surfactant for the degradation of phenanthrene by Polyporus sp. S 133. The degradation rate increased as the amount of Tween 80 added increased. The rate in agitated cultures was about 2 times that in non-agitated cultures. The mechanism of degradation was determined through the identification of metabolites; 9,10-phenanthrenequinone, 2,2＇-diphenic acid, phthalic acid, and protocatechuic acid. Several enzymes （manganese peroxidase, lignin peroxidase, laccase, 1,2-dioxygenase and 2,3-dioxygenase） produced by Polyporus sp. S133 were detected during the incubation. The highest level of activity was shown by 1,2-dioxygenase （187.4 U/L） after 20 days of culture.

Uptake and accumulation of phenanthrene and pyrene in spiked soils by Ryegrass(Lolium perenne L.)

Phytoremediation has long been recognized as a cost-effective method for the removal of polycyclic aromatic hydrocarbons （PAHs） from soil. A study was conducted to investigate the uptake and accumulation of PAHs in root and shoot of Lolium perenne L. Pot experiments were conducted with series of concentrations of 3.31-378.37 mg/kg for phenanthrene and those of 4.22-365.38 mg/kg for pyrene in a greenhouse. The results showed that both ryegrass roots and shoots did take up PAHs from spiked soils, and generally increased with increasing concentrations of PAH in soil. Bioconcentration factors（BCFs） of phenanthrene by shoots and roots were 0.24- 4.25 and 0.17-2.12 for the same treatment. BCFs of pyrene by shoots were 0.20-1.5, except for 4.06 in 4.32 mg/kg treatment, much lower than BCFs of pyrene by roots （0.58-2.28）. BCFs of phenanthrene and pyrene tended to decrease with increasing concentrations of phenanthrene and pyrene in soil. Direct uptake and accumulation of these compounds by Lolium perenne L. was very low compared with the other loss pathways, which meant that plant-promoted microbial biodegradation might be the main contribution to plant-enhanced removal of phenanthrene and pyrene in soil. However, the presence of Lolium perenne L. significantly enhanced the removal of phenanthrene and pyrene in spiked soil. At the end of 60 d experiment, the extractable concentrations of phenanthrene and pyrene were lower in planted soil than in non-planted soil, about 83.24%-91.98% of phenanthrene and 68.53%-84.10% of pyrene were removed from soils, respectively. The results indicated that the removal of PAHs in contaminated soils was a feasible approach by using Lolium perenne L.

Phytotoxicity assessment of phenanthrene, pyrene and their mixtures by a soil-based seedling emergence test

Seedling emergence tests were conducted in a meadow brown soil using five plant species(i.e., Chinese cabbage, green onion, tomato, turnip and wheat) to determine the phytotoxicity of phenanthrene, pyrene and their mixtures. The soil was amended with up to 1000 mg/kg soil of phenanthrene or 600 mg/kg soil of pyrene. Seedling emergence and root growth were measured. The results indicated that root elongation was more sensitive than seedling emergence. Root length decreased with increasing phenanthrene or pyrene concentrations(p≤0 05). Phenanthrene was more phytotoxic than pyrene. The lowest observable adverse effect concentration(LOAEC) was 10 mg/kg for phenanthrene when tested with green onion, and 50 mg/kg for pyrene when tested with wheat. Among the five species, wheat was found to be the most sensitive. When amended jointly at or below their respective LOAEC, phenanthrene and pyrene produced a synergistic toxic effect.

Characterization and phylogenetic analysis of a phenanthrene-degrading strain isolated from oil-contaminated soil

Bacterium strain EVA17 was isolated from an oil-contaminated soil, and identified as Sphingomonas sp. based on analysis of 16S rDNA sequence,cellular fatty acid composition and physiological-chemical tests. The salicylate hydroxylase and catechol 2,3-dioxygenase(C23O) were detected in cell-free lysates, suggesting a pathway for phenanthrene catabolism via salicylate and catechol. Alignment showed that both of the C23O and GST genes of the strain EVA17 had high similarity with homologues of strains from genus Sphingomonas. The phylogenetic analysis based on 16S rDNA and C23O gene sequence indicated that EVA17 should be classified into genus Sphingomonas, although the two phylogenetic trees were slightly different from each other. The results of co-amplification and sequence determination indicated that GST gene should be located upstream of the C23O gene.

Kinetics study of aqueous sorption of phenanthrene to humic acids and sediments

The sorption behavior was determined for a model polycyclic aromatic hydrocarbon(PAH), i.e., phenanthrene(PHN), from water to three humic acids(HAs) and three sediments in different reacting time. The chemical compositions of HA samples were measured using cross polarization magic angle spinning carbon 13(CPMAS 13 C NMR along with elemental analysis. The dissolved humic substances dissociating from solid HAs and sediments were characterized by 1 H NMR. The experiments indicated that the sorption modes and mechanisms of natural sorbents for PHN varied significantly between short(<7 d) and long contact time and the reaction time should be taken into consideration in studying the overall sorption process. The sorption capacity( K ′ f) and exponent( n ) might be relative to the properties of dissolved humic materials in initial stage but the solid aromatic organic matter after long time reaction. According to the experiments performed in this investigation and the previous researches, a conceptive sorption model was established.

Isolation and Characteristics of a Microbial Consortium for Effectively Degrading Phenanthrene

A microbial consortium （named W4） capable of aerobic biodegradation of solid phenanthrene as the sole source of carbon and energy was isolated by selective enrichment from petroleum-contaminated soil in the Henan oilfield, China. The strains of the consortium were identified as Sphingomonas cloacae, Rhizobium sp., Pseudomonas aeruginosa and Achromobacter xylosoxidans respectively by means of genetic methods. The major metabolites of phenanthrene were analyzed by gas chromatography-mass spectrometry （GC-MS）. The biodegradation percentage of solid phenanthrene at 200 mg/L in liquid medium after 7 days of growth was greater than 99%. The degradation of phenanthrene was compared between individual predominant strains and the microbial consortium in different treatment processes. The microbial consortium showed a significant improvement of phenanthrene degradation rates in either static or shaking culture. The degradation percentage of phenanthrene by the consortium W4 decreased to some degree when C 16 coexisted, however it was hardly affected by C30. Furthermore, the ability of consortium W4 to remediate oil sludge from the Dagang oil refinery was studied by composting, and it was found that the consortium W4 could obviously remove polycyclic aromatic hydrocarbons （PAils） and paraffinic hydrocarbons. All the results indicated that the microbial consortium W4 had a promising application in bioremediation of oil-contaminated environments and could be potentially used in microbial enhanced oil recovery （MEOR）.

Kinetics and characteristics of phenanthrene degradation by a microbial consortium

The kinetics and characteristics of phenanthrene degradation by a microbial consortium W4 isolated from Henan Oilfield were investigated. The degradation percentage of solid phenanthrene at 200 mg/L in liquid medium after 6 days of incubation was higher than 95% under the condition of 37 ℃ and 120 r/min by this microbial consortium. The degradation of phenanthrene could be fitted to a first-order kinetic model with the half-life of 1.25 days. The optimum conditions for degradation ofphenanthrene by consortium W4 were as follows： temperature about 37℃, pH from 6.0 to 7.0 and salinity about 8.0 g/L. It was concluded that microbial consortium W4 might degrade phenanthrene via both salicylic acid and o-phthalic acid pathways by analyzing products with GC-MS.

Thermodynamics of phenanthrene partition into solid organic matter from water

The thermodynamic behavior of organic contaminants in soils is essential to develop remediation technologies and assess risk from alternative technologies. Thermodynamics of phenanthrene partition into four solids(three soils and a bentonite) from water were investigated. The thermodynamics parameters(ΔH, ΔG o, ΔS o,) were calculated according to experimental data. The total sorption heats of phenanthrene to solids from water ranged from -7.93 to -17.1 kJ/mol, which were less exothermic than the condensation heat of phenanthrene-solid(i.e., -18.6 kJ/mol). The partition heats of phenanthrene dissolved into solid organic matter ranged from 23.1 to 32.2 kJ/mol, which were less endothermic than the aqueous dissolved heat of phenanthrene(i.e., 40.2 kJ/mol), and were more endothermic than the fusion heat of phenanthrene-solid(i.e., 18.6 kJ/mol). The standard free energy changes, ΔG o, are all negative which suggested that phenanthrene sorption into solid was a spontaneous process. The positive values of standard entropy changes, ΔS o, show a gain in entropy for the transfer of phenanthrene at the stated standard state. Due to solubility-enhancement of phenanthrene, the partition coefficients normalized by organic carbon contents decrease with increasing system temperature(i.e., lnK oc =-0.284lnS+9.82(n=4, r 2=0.992)). The solubility of phenanthrene in solid organic matter increased with increasing temperatures. Transports of phenanthrene in different latitude locations and seasons would be predicted according to its sorption thermodynamics behavior.

Alkyl Naphthalenes and Phenanthrenes:Molecular Markers for Tracing Filling Pathways of Light Oil and Condensate Reservoirs

Condensates and light oils are generally characterized by high maturity, low concentration of sterane and terpane biomarkers and low content of non-hydrocarbon fraction. As a result, some commonly-used sterane, terpane and carbazole migration parameters in conventional oil reservoirs may have a certain limitation in condensate and light oil reservoirs for their poor signal-noise ratios in the gas chromatography-mass spectrometry （GC-MS）. Naphthalene, phenanthrene and their methylated substituents, however, are present in significant concentrations in condensates and light oils. Taking the Fushan depression （in the Beibuwan Basin, Northern South China Sea） as an example, this paper attempts for the first time to use polycyclic aromatic hydrocarbon （PAH）-related parameters to trace migration directions and filling pathways for condensate and light oil reservoirs. The result shows that TMNr （i.e. 1, 3, 7-TMN/（1, 3, 7-TMN ＋ 1, 2, 5-TMN）, TMN： trimethylnaphthalene））, MPI-1 （i.e. 1.5×（2-MP ＋ 3-MP）/（P ＋ 1-MP ＋ 9-MP）, P： phenanthrene MP： methyiphenanthrene）, MN/DMN （∑methylnaphthalene/∑dimethylnaphthalene, %） and MN/TMN （∑nethylnaphthalene/∑trimethylna- phthalene, %） can be used to trace the filling pathways of condensate and light oil reservoirs. These parameters, together with geological consideration and other bulk oil properties （e.g. the gas to oil ratio and density）, suggest that the condensates and light oils in the Huachang oil and gas field are mainly sourced from the Bailian sag that is located to the northeast of the Huachang uplift in the Fushan depression.

Effect of biosurfactant on the sorption of phenanthrene onto original and H_2O_2-treated soils

The objective of this study was to examine the effect of biosurfactant on the sorption of phenanthrene （PHE） onto the original or HzO2-treated black loamy soil （typic isohumisols） and red sandy soil （typic ferralisols）. The sorption isotherms were performed with the original and ＂soft＂ carbon-removed soils in the presence and absence of biosurfactant （200 mg/L）. The sorption and degradation of biosurfactant were investigated. The result showed that organic matter played an important role in PHE sorption onto the black loamy and red sandy soils, and the PHE sorption isotherms on the ＂soft＂ carbon-removed soils exhibited more nonlinearity than those on the original soils. The values of partition coefficient （Kd） on the original black loamy soil with or without 200 mg/L biosurfactant were 181.6 and 494.5 mL/g, respectively. Correspondingly, in the red sandy soil, Kd was 246.4 and 212.8 mL/g in the presence or absence of biosurfactant, respectively. The changes of Kd suggested that biosurfactant inhibited PHE sorption onto the black loamy soil, but facilitated PHE sorption onto the red sandy soil. The nonlinearity of PHE sorption isotherm was decreased in the presence of biosurfactant. Site specific sorption might occur during PHE sorption onto both the original and the ＂soft＂ carbon-removed soils in the presence of biosurfactant. It was noted that biosurfactant could also be sorbed onto soils. The maximal sorption capacity of the red sandy soil for biosurfactant was （76.9 ± 0.007） μg/g, which was 1.31 times that of black loamy soil. Biosurfactant was degraded quickly in the two selected soils, and 92% of biosurfactant were mineralized throughout the incubation experiment for 7 d. It implied that biosurfactant should be added frequently when the remediation of polycyclic aromatic hydrocarbon （PAH）-contaminated soils was conducted through PAH desorption approach facilitated by biosurfactant.

Effects of humic acid coatings on phenanthrene sorption to black carbon

Black carbon （BC） can strongly adsorb hydrophobic organic compounds （HOCs）. The HOC sorption to coated BC could be attenuated in soil and sediment compared with that of the parent BC. To study the potential causes of the sorption attenuation, humic acid （HA） and BC were isolated. Phenanthrene （PHE） was selected as the representative of HOCs. BC was coated with the precipitated HA. The PHE sorption to the HA-coated BC was determined. The HA coatings on BC could result in the significant sorption attenuation of PHE to BC. The attenuation varied in different HA origin and was positively correlated to the aromaticity of HA. The attenuation could be explained by the direct competition between HA and PHE for the available sorption sites on BC and the reduction of the available sorption sites as a result of the pore blockage of BC caused by the HA coatings. Therefore, the HA coatings on BC was one potential cause of the attenuation of HOC sorption to BC in soil and sediment.