Molecular characterization of phenanthrene-degrading methanogenic communities in leachate-contaminated aquifer sediment

The existence of polycyclic aromatic hydrocarbons in the various environments has aroused great environmental concerns due to their potential hazards to human health. The presence of polycyclic aromatic hydrocarbons in aquifer is particularly sensitive where groundwater is used as a source of potable water. Anaerobic biodegradation of polycyclic aromatic hydrocarbons is an attractive option for remediation of contaminated aquifer sediment. Bacterial and archaeal community structures of phenanthrene-degrading aquifer sediment under methanogenic condition were investigated using clone library analysis in combination with microcosm study. The bacterial members were all affiliated with ??-Proteobacteria. Phylum Euryarchaeota was the predominant archaeal group, represented by genera Methanosarcina, Methanobacterium and Thermogymnomonas. Both bacteria (genera Citrobacter and Pseudomonas) and archaea (genus Methanosarcina) might have links to phenanthrene degradation process. This work might provide some new insights into developing strategies for the isolation of the putative polycyclic aromatic hydrocarbons degraders under methanogenic condition and bioremediating polycyclic aromatic hydrocarbons in leachate-contaminated aquifer sediment.     

Solvent–water extraction method for the evaluation of polycyclic aromatic hydrocarbons bioavailability in coal–tar-contaminated soils

A solvent–water extraction method was proposed as an assessment tool to estimate the bioavailability of polycyclic aromatic hydrocarbons in coal–tar-contaminated soils. The approach taken was to measure the percent of polycyclic aromatic hydrocarbons extracted by a solvent–water mixture and comparing the results with the percent of polycyclic aromatic hydrocarbons degraded in a soil slurry reactor. Five soil samples from three former manufactured gas plant sites and a coal–tar disposal site which were operated between 1880 and 1947, and 1945 and 1950, respectively, in Iowa, USA were used in this study. Extraction experiments were conducted using acetone–water or ethanol–water mixtures with solvent volume fractions ranging from 1.0 to 0.4 (v/v). The percent of polycyclic aromatic hydrocarbons extracted from the various soils decreased as the volume fraction of the solvent in the solvent–water mixture was reduced. An acetone–water mixture of 0.6 was found to be appropriate in correlating the percent of polycyclic aromatic hydrocarbons degraded to the percent of polycyclic aromatic hydrocarbons extracted. For the first correlation, the percent extracted and the percent biodegraded were modified by using the molecular weights and log K _ow of polycyclic aromatic hydrocarbons, respectively. For the second correlation, the equation relating the percent extracted and the percent biodegraded was modified using soil properties such as organic carbon content and percent of clay and silt. Although the experiments were conducted for a limited number of soils, the extraction method appeared to be a good starting point in estimating the bioavailability of polycyclic aromatic hydrocarbons in coal–tar-contaminated soils.     

Biodegradation of polycyclic aromatic hydrocarbon by a halotolerant bacterial consortium isolated from marine environment

The biodegradability of polycyclic aromatic hydrocarbons such as naphthalene, fluorene, anthracene and phenanthrene by a halotolerant bacterial consortium isolated from marine environment was investigated. The polycyclic aromatic hydrocarbons degrading bacterial consortium was enriched from mixture saline water samples collected from Chennai (Port of Chennai, salt pan), India. The consortium potently degraded polycyclic aromatic hydrocarbons (> 95%) at 30g/L of sodium chloride concentration in 4 days. The consortium was able to degrade 39 to 45% of different polycyclic hydrocarbons at 60 g/L NaCl concentration. Due to increase in salinity, the percent degradation decreased. To enhance polycyclic aromatic hydrocarbons degradation, yeast extract was added as an additional substrate at 60g/L NaCl concentration. After the addition of yeast extract, the consortium degraded > 74 % of polycyclic aromatic hydrocarbons at 60 g/L NaCl concentration in 4 days. The consortium was also able to degrade PAHs at different concentrations (5, 10, 20, 50 and 100 ppm) with 30 g/L of NaCl concentration. The polycyclic aromatic hydrocarbons degrading halotolerant bacterial consortium consists of three bacterial strains, namely Ochrobactrum sp., Enterobacter cloacae and Stenotrophomonas maltophilia.     

Hydrocarbon biodegradation and soil microbial community response to repeated oil exposure

Bioremediation strategies continue to be developed to mitigate the environmental impact of petroleum hydrocarbon contamination. This study investigated the ability of soil microbiota, adapted by prior exposure, to biodegrade petroleum. Soils from Barrow Is. (W. Australia), a class A nature reserve and home to Australia’s largest onshore oil field, were exposed to Barrow production oil (50 ml/kg soil) and incubated (25 °C) for successive phases of 61 and 100 days. Controls in which oil was not added at Phase I or II were concurrently studied and all treatments were amended with the same levels of additional nutrient and water to promote microbial activity. Prior exposure resulted in accelerated biodegradation of most, but not all, hydrocarbon constituents in the production oil. Molecular biodegradation parameters measured using gas chromatography–mass spectrometry (GC–MS) showed that several aromatic constituents were degraded more slowly with increased oil history. The unique structural response of the soil microbial community was reflected by the response of different phospholipid fatty acid (PLFA) sub-classes (e.g. branched saturated fatty acids of odd or even carbon number) measured using a ratio termed Barrow PLFA ratio (B-PLFAr). The corresponding values of a previously proposed hydrocarbon degrading alteration index showed a negative correlation with hydrocarbon exposure, highlighting the site specificity of PLFA-based ratios and microbial community dynamics. B-PLFAr values increased with each Phase I and II addition of production oil. The different hydrocarbon biodegradation rates and responses of PLFA subclasses to the Barrow production oil probably relate to the relative bioavailability of production oil hydrocarbons. These different effects suggest preferred structural and functional microbial responses to anticipated contaminants may potentially be engineered by controlled pre-exposure to the same or closely related substrates. The bioremediation of soils freshly contaminated with petroleum could benefit from the addition of exhaustively bioremediated soils rich in biota primed for the impacting hydrocarbons.     

Promoted dissipation of phenanthrene and pyrene in soils by amaranth (<Emphasis Type="Italic">Amaranthus tricolor</Emphasis> L.)

A study was conducted to investigate the performance of amaranth, a known hyperaccumulator of cesium, on the promotion of the dissipation of soil phenanthrene and pyrene, which are PAHs (polycyclic aromatic hydrocarbons). Amaranthus tricolor L. een choi was the cultivar used. The presence of Amaranthus tricolor L. evidently enhanced the dissipation of these PAHs in soils with initial phenanthrene concentrations of 7.450–456.5 mg/kg dw (dw, dry weight) and pyrene of 8.010–488.7 mg/kg dw. At the end of the experiment (45 days), the residual concentrations of phenanthrene and pyrene in spiked soils with plants were generally higher than those with no plants. The loss of phenanthrene and pyrene in vegetated soils was 87.85–94.03% and 46.89–76.57% of the soil with these chemicals, which was 2.55–13.66% and 11.12–56.55% larger than the loss in non-vegetated soils, respectively. The accumulation of phenanthrene and pyrene by the plant was evident. Root and shoot concentrations of these chemicals monotonically increased with increasing soil PAH concentrations. Bioconcentration factors (BCFs), defined as the ratio of chemical concentrations in plants and in the soils (on a dry weight basis), of phenanthrene and pyrene by roots were 0.136–0.776 and 0.603–1.425, while by shoots were 0.116–0.951 and 0.082–0.517, respectively. BCFs of phenanthrene and pyrene tended to decrease with the increasing concentrations of soil phenanthrene and pyrene. Plant accumulation only accounted for less than 0.32% (for phenanthrene) and 0.33% (for pyrene) of the total amount enhancement of the dissipated PAHs in vegetated vs. non-vegetated soils. In contrast, plant-promoted microbial biodegradation was the predominant contribution to the plant-enhanced dissipation of soil phenanthrene and pyrene. These results suggested the feasibility of the radionuclide hyperaccumulator in phytoremediating the soil PAH contaminants.     

Enhanced biodegradation of polycyclic aromatic hydrocarbons using nonionic surfactants in soil slurry

The effect of nonionic surfactants on the solubility and biodegradation of polycyclic aromatic hydrocarbons (PAHs) in the aqueous phase and in the soil slurry phase, as well as the fate of these surfactants, were investigated. The PAH solubility was linearly proportional to the surfactant concentration when above the critical micelle concentration (CMC), and increased as the hydrophile–lipophile balance (HLB) value decreased. Substantial amounts of the sorbed phenanthrene in the soil particles were desorbed by non-ionic surfactants into the liquid phase when the ratio of soil to water was 1:10 (g/ml). Brij 30 was the most biodegradable surfactant tested, showed no substrate inhibition up to a concentration of 1.5 g/l, and was definitely used as a C source by the bacteria. Naphthalene and phenanthrene were completely degraded by phenanthrene-acclimatised cultures within 60 h, but a substantial amount of naphthalene was lost due to volatilization. The limiting step in the soil slurry bioremediation was bioavailability by the micro-organisms for the sand slurry and mass transfer from a solid to aqueous phase in the clay slurry.     

Pollution characteristics of polycyclic aromatic hydrocarbons （PAHs） in oily sludge from the Zhongyuan Oilfield and its peripheral soils

The purpose of this study is to determine the degree of contamination caused by polycyclic aromatic hydrocarbons (PAHs) in oily sludge and soils around it in the Zhongyuan Oilfield. The contents of polycyclic aromatic hydrocarbons in oily sludge samples were determined with HPLC. The contents of PAHs of oily sludge from three different oil production plants vary from high to low in the order of the Wenming oily sludge dumping site of No. 3 Oil Production Plant (3W)>the Mazhai oily sludge dumping site of No. 3 Oil Production Plant (3M)>the Wen’er oily sludge dumping site of No. 4 Oil Production Plant (4W). Naphthalene, acenaphthylene, acenaphthene, fluorine and phenanthrene are the major pollutants of PAHs in oily sludge. The contents of PAHs in soil samples around the oily sludge dumping sites vary widely from 434.49 to 2408.8 ng/g. Naphthalene, acenaphthene, fluorine, phenanthrene and pyrene are the characteristic factors of PAHs in soil samples of 3M and 3W, and naphthalene, acenaphthene, fluorine and phenanthrene are the characteristic factors of PAHs in soil samples of 4W. According to these data and the ratios of Fl/Py, PAHs in oily sludge samples come mainly from petrogenic sources, and soil samples are divided into petrogenic soil samples and mixed-source soil samples, and both petrogenic and pyrogenic soil samples in terms of the sources of PAHs. The classification by Nemero index P indicates that soils around the oily sludge dumping sites have been seriously polluted.     

Bioremediation of heavily hydrocarbon-contaminated drilling wastes by composting

Oil-based drilling cuttings comprise a large and hazardous waste stream generated by oil and gas wells drilling operations. Oil-based cuttings are muddy materials with high contents of salts and hydrocarbons. Composting strategies have shown to be effective in the biodegradation of petroleum hydrocarbons, and it offers numerous advantages in comparison with other bioremediation methods. In order to assess the effectiveness of drilling cuttings bioremediation by composting with food and garden wastes, an experiment was conducted in 60-L reactors for 151 days. Four treatments were carried out: only oil-based cuttings, two proportions (in a volume basis) of organic wastes to drilling cuttings (33 and 75 %) and only organic wastes (as a traditional composting reference), with pine-tree woodchips as bulking agent. High degradation percentages of total hydrocarbons (≈82 %), n-alkanes (≈96 %) and the 16 USEPA-listed polycyclic aromatic hydrocarbons (≈93 %) were reached in the treatment with 75 % of organic wastes, and applying 33 % of organic wastes was not more effective than not applying organic wastes for the drilling cuttings hydrocarbons biodegradation. Furthermore, in the treatment with 75 % of organic wastes, alkanes half-life and polycyclic aromatic hydrocarbons half-life were about 10 times and four times lower, respectively, than those in the treatment with 33 % of organic wastes. Possibly, lower hydrocarbons and salts initial concentrations (i.e., lower toxicity), higher microbial counts, adequate nutrient proportions and water content supported a high biological activity with a consequent elevated biodegradation rate in the treatment with 75 % of organic wastes.     

Effect of soil structure on the bioavailability of polycyclic aromatic hydrocarbons within aggregates of a contaminated soil

Biodegradation of polycyclic aromatic hydrocarbon (PAH) was investigated in the whole matrix and in the different aggregate size fractions of a sandy soil contaminated by a mixture of 8 PAHs and incubated at water holding capacity. The distribution of PAHs and of phenanthrene-degrading bacteria were determined in the bulk soil and in 4 size aggregate fractions corresponding to sand, coarse silt, fine silt and clay. The microbial communities able to degrade phenanthrene were detected at a similar level in the different aggregate fractions of the soil before contamination. After soil contamination and incubation, a significant growth of bacteria was observed and their distribution within aggregates was modified. Bacterial communities of phenanthrene-degraders were present in a higher density in the aggregates corresponding to sand (2000–50 μm) and clay (<2 μm). Chemical analysis show that remaining PAHs (low and high molecular weight) were much more concentrated in the fine soil fractions (fine silt and clay) and were present at a very low content in the larger aggregate size fractions. The interactions of well defined aggregates with PAHs and bacteria were also studied using phenanthrene as PAH model substrate and individual aggregates corresponding to sand and clay size fractions. Incubation of sand and clay aggregate fractions enriched with phenanthrene in the presence of a bacterial isolate NAH1 led to the simultaneous solubilization and biodegradation of phenanthrene. Differences in amounts of solubilized phenanthrene between sand and clay aggregate size fractions would be related to difference in adsorption capacities of phenanthrene by clay and sand aggregates.     

Polycyclic aromatic hydrocarbons in the soils of technogenic landscapes

An integrated study of qualitative and quantitative composition of polycyclic aromatic hydrocarbons (PAH) in the atmospheric precipitation-soil-lysimetric water system of aerotechnogenic polluted landscapes was conducted using high-performance liquid chromatography in a gradient mode. Only low-molecular weight polyarenes (phenanthrene, anthracene, fluoranthene, pyrene, benz(a)anthracene, and chrysene) were found in the atmospheric precipitation and lysimetric waters. The growth of PAHs in soils is provided by the input of phenanthrene, fluoranthene, and pyrene with atmospheric precipitation. The absence of heavy PAHs (benzfluoranthenes, benz(a)pyrene, dibenz(a,h)anthracene, benz(ghi) perylene, and indeno[1,2,3-cd]pyrene) in the atmospheric precipitation and their identification in soil give grounds to state that their accumulation was caused mainly by transformation of organic matter during pedogenesis. The technogenic impact was estimated and criterion of the degree of soil pollution by PAH was proposed.     

Specific detection of bioavailable phenanthrene and mercury by bacterium reporters in the red soil

Genetically engineered Pseudomonas putida reporters (BMB-PL and BMB-ME), which, respectively, carried phnS-luxCDABE and merR-egfp cassette, were used to determine bioavailable phenanthrene and mercury. Over a spiked range of concentrations and aged for 6 days in red soil samples, the reporters were tested to determine the optimal assay conditions and the bioavailable phenanthrene (0–60 mg kg^?1) and Hg²⁺ (0–240 μg kg^?1) were evaluated by the signal of the relative fluorescent units and relative luminescence units. Single contamination was carried out and good correlations were obtained between signal strength and pollutant concentrations, whereas interference and bioavailability repression were observed in dual-contamination experiments. Other heavy metal ions at nanomolar level did not interfere with BMB-ME measurement while BMB-PL showed some response to other polycyclic aromatic hydrocarbons or their intermediate products during degradation. Comparing high-performance liquid chromatography methods with the bacterial reporters, both BMB-ME and BMB-PL appeared to have a detection limit (mercury <40 μg kg^?1; phenanthrene <24 mg kg ^?1) similar to the instrumental analysis. Although physical parameters may affect the interaction of pollutants with bioreporter cells, advantages include the inherent biological relevance of the response, rapid response time, and potential for field deployment. Our results strongly suggest that the BMB-ME and BMB-PL bioreporters constitute an adaptable system for easily detecting the bioavailability of mercury and phenanthrene in the red soils.     

Evidence for in situ methanogenic oil degradation in the Dagang oil field

In situ biotransformation of oil to methane was investigated in a reservoir in Dagang, China using chemical fingerprinting, isotopic analyzes and molecular and biological methods. The reservoir is highly methanogenic despite chemical indications of advanced oil degradation, such as depletion of n-alkanes, alkylbenzenes and light polycyclic aromatic hydrocarbon (PAH) fractions or changes in the distribution of several alkylated polycyclic aromatic hydrocarbons. The degree of degradation strongly varied between different parts of the reservoir, ranging from severely degraded to nearly undegraded oil compositions. Geochemical data from oil, water and gas samples taken from the reservoir are consistent with in situ biogenic methane production linked to aliphatic and aromatic hydrocarbon degradation. Microcosms were inoculated with production and injection waters in order to characterize these processes in vitro. Subsequent degradation experiments revealed that autochthonous microbiota are capable of producing methane from ¹³C labelled n-hexadecane or 2-methylnaphthalene and suggest that further methanogenesis may occur from the aromatic and polyaromatic fractions of Dagang reservoir fluids. The microbial communities from produced oil–water samples were composed of high numbers of microorganisms (on the order to 10⁷), including methane producing Archaea within the same order of magnitude. In summary, the investigated sections of the Dagang reservoir may have significant potential for testing the viability of in situ conversion of oil to methane as an enhanced recovery method and biodegradation of the aromatic fractions of the oil may be an important methane source.     

拉萨市拉鲁湿地多环芳烃污染及其来源

通过分析采集的土壤、沉积物和大气样品,对拉萨郊区的拉鲁湿地多环芳烃污染及其来源进行了初步研究.湿地表层土壤中16种多环芳烃的平均含量的总含量为82.45×10^-9,既存在低环数的多环芳烃,又含有高环数的多环芳烃;湿地土壤中的多环芳烃主要来自于大气.     

Application of indigenous microbial consortia in bioremediation of oil-contaminated soils

Bioremediation of oil spillage in soils using consortia of microbes beckons much exploration. The present study involves bioremediation of oil-contaminated soils from north Chennai, India, using indigenous microbial consortia. Totally, 32 positive oil degrading isolates were obtained from 3 different locations, i.e., petrol filling stations, automobile workshops and oil refineries. Substrate utilization patterns of individual isolates and the consortial sets were observed. Mixture of three common hydrocarbons (petrol, diesel and engine oil) was used for studies. The substrate oil utilized by consortia was taken for thin-layer and column chromatography which perfectly resulted in varied fractions of oil compared to the unused oil as control. The best consortia were used directly for bioremediation experiment. Three different oil-contaminated soils were used and bioremediation patterns were observed. The rate of bioremediation differed within soils, nevertheless all soils were almost 100 % reclaimed within 30 days. Bioremediation kinetics showed that the process corresponds to first-order kinetics and kinetic constants for the different soils ranged from 0.051 to 0.077/day. Assessment of detoxification of acute phytotoxicity owing to the pollutant oil was done, and results observed were significant. An increase of 25, 300 and 212 % in germination index, average growth index and sustenance index, respectively, of Trigonella foenum-graecum Linn. in treated soils was observed, compared to untreated soils. Thus, this study confirmed that microbes in ‘Consortial Union’ serve as better treating agents in bioremediation of oil-contaminated soils than individual microorganisms.     

Development of a two-stage washing and biodegradation system to remediate octachlorinated dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin-contaminated soils

A two-stage system for octachlorinated dibenzo-p-dioxin (OCDD)-contaminated soil remediation was developed. Soil washing using emulsified oil (EO) was applied in the first stage for OCDD extraction followed by the second stage of bioremediation using P. mendocina NSYSU for remaining OCDD biodegradation. The major tasks included (1) determination of optimal soil washing conditions for OCDD extraction by EO, (2) evaluation of feasibility of OCDD biodegradation by P. mendocina NSYSU under aerobic cometabolic conditions using EO as the primary substrate, and (3) assessment of the effectiveness of OCDD removal using the two-stage system. During the soil washing stage, EO with two different oil-to-water ratios (1:50 and 1:200) and pore volumes were tested with initial soil OCDD concentration of 21,000 µg/kg. Results indicate that EO could effectively improve the solubility and desorption of OCDD in soils. Up to 74% of OCDD removal could be obtained after washing with 60 PVs of EO and dilution factor of 50. After the soil washing process, enriched P. mendocina NSYSU solution was added into the reactor to enhance the aerobic biodegradation of remaining OCDD in soils. P. mendocina NSYSU could use adsorbed EO globules as substrates and caused significant OCDD degradation via the aerobic cometabolic mechanism. Approximately 82% of the remaining OCDD could be removed after 50 days of operation, and P. mendocina NSYSU played important roles in OCDD biodegradation. Up to 87% of OCDD was removed through the EO washing and biodegradation process. The two-stage system is a potential technology to remediate dioxin-contaminated soils.     

Sorption kinetics of naphthalene and phenanthrene in loess soils

A laboratory study was executed to investigate the effect of surfactants to enhance sorption of polycyclic aromatic hydrocarbon (PAH) contaminants in loess soil. Phenanthrene and naphthalene were chosen as organic contaminant indicators in loess soil modified by the cation surfactant hexadecyltrimethylammonium (HDTMA) bromide. The kinetic behavior of sorption during transport in natural and modified loess soil was studied. The results indicated that sorption rate in the cation surfactant modified loess soils was at least 3 times faster than that of the natural soil. A first-order kinetics model fitted the sorption data well for both soils. The sorption rates of the two organic compounds were related to their primary residual quantity on the soils. The experiments showed that sorption amounts approached constant values approximately within 30 and 90 min for naphthalene and phenanthrene at 298–318 K, respectively. The rate constants, however, displayed negative correlation with increasing temperature. With changing temperature, the activation energy was calculated at –6.196–1.172 kJ/mol for naphthalene and –28.86–15.70 kJ/mol for phenanthrene at 298–318 K. The results can be used to predict the sorption kinetics of phenanthrene and naphthalene in loess soils, and in a wider perspective, be used to better understand the transport of petroleum contaminants in the soil environment.     

Development of a thermal desorption method for the analysis of particle associated polycyclic aromatic hydrocarbons in ambient air

An investigation for the analysis of polycyclic aromatic hydrocarbons in airborne particulates using thermal desorption and gas chromatography-mass spectrometry is described. Samples are obtained from ambient air using fibreglass filters and the volatile material from the filter is thermally desorbed to gas chromatograph. A 30 meter capillary column is used to separate the hydrocarbons and eight polyaromatic hydrocarbons are used to test the method and recovery is >95%. The eight polycyclic aromatic hydrocarbons anthracene, phenanthrene, fluoranthrene, pyrene, benzo (a) anthracene, chrysene, benzo (a) pyrene and benzo (e) pyrene were the most abundant PAHs found in the samples of ambient air with current method at Uxbridge-London. Application of the measurement of polycyclic aromatic hydrocarbons in ambient air samples shows that the hydrocarbons trapped in the particle phase to a lesser degree at higher ambient temperature. In conclusion a method has been developed to transfer the PAHs in particle phase from a filter to GC-MS by thermal desorption. A standard mixture of PAHs, when absorbed onto the filter, did lead to strong analyte absorbent interactions by the high percent recovery of the sample.     

Evaluation of the diagnostic ratios for the identification of spilled oils after biodegradation

Biodegradation, one of the most important weathering processes, alters the composition of spilled oil, making it difficult to identify the source of the release and to monitor its fate in the environment. A laboratory experiment was conducted to simulate oil spill weathering process of microbial degradation to investigate compositional changes in a range of source- and weathering-dependent molecular parameters in oil residues, and the conventional diagnostic ratios for oil spill identification were also evaluated. The conventional diagnostic ratios of n-alkane displayed obvious changes after biodegradation, especially for Pr/n-C₁₇ and Ph/n-C₁₈ with relative standard deviation more than 118.84 %, which suggests they are invalid for oil source identification of the middle-serious spill. Many polycyclic aromatic hydrocarbons (PAHs) are more resistant to biodegradation process than their saturated hydrocarbon counterparts, thus making PAHs to be one of the most valuable fingerprinting classes of hydrocarbons for oil identification. Biomarker ratios of hopanes and steranes were also useful for source identification even after moderate biodegradation, and the diagnostic ratios from them could be used in tracking origin and sources of hydrocarbon pollution. Finally, the carbon isotopic type curve may provide another diagnostic means for correlation and differentiation of spilled oils, and be particularly valuable for lighter refined products or severely biodegraded oils, the source of which may be difficult to identify by routine biomarker techniques.     

Estimation of bioavailability of polycyclic aromatic hydrocarbons in river sediments

This study aimed to evaluate the total concentration of polycyclic aromatic hydrocarbons in sediments of Iguassu River in Southern Brazil. Alongside the concentration, the amount of such compounds bioavailable was also evaluated. This is accomplished by comparing its total amount present in sediments and the amount extracted by n-butanol. The results showed that the total concentration of polycyclic aromatic hydrocarbons presented in sediment ranged from 4.49 to 58.75???g/g. The total amount of polycyclic aromatic hydrocarbons extracted by n-butanol ranged from 1.22 to 17.07???g/g. The use of n-butanol represents the mimetic conditions that hydrocarbons, derived from oil, could be taken up by organisms. Most of the hydrocarbons extracted by n-butanol were those with lower octanol?Cwater partition constant, usually those with three and four rings. Compounds with more than four rings were extracted in lower or insignificant amounts. Even the hydrocarbons with lower molecular weight available may be degraded or eliminated by organisms, when accumulated. Estimating bioavailability of hydrocarbons represents what specific hydrocarbons could be available to be taken up by organisms.     

Combined effects of heavy metal and polycyclic aromatic hydrocarbon on soil microorganism communities

Soil contaminated sites contain a variety of pollutants, especially heavy metals and polycyclic aromatic hydrocarbons (PAHs). Interactions between heavy metals have been relatively well studied, but little is known about interactions between heavy metals and PAHs. The combined effect of heavy metals and PAHs on soil microorganism was studied in laboratory conditions and evaluated by random denaturing gradient gel electrophoresis. We extracted DNA directly from contaminated soils and then amplified the V3 sequences of the 16S rDNA. The results showed that with different culture time, the gene diversity of the single and combined contaminated soil differed as well. After 15 days of culture, the microorganisms were stimulated and accommodated. After 45 days of cultivation, the quantities of the soil microorganisms were affected. It is concluded that some of the microorganisms utilize phenanthrene as important carbon resources. Microorganisms directly isolated from soil could reflect the diversity of soil microorganism and population distribution conditions.