Metagenomic analysis reveals the microbial response to petroleum contamination in oilfield soils.

The response of the microbes to total petroleum hydrocarbons (TPHs) in three types of oilfield soils was researched using metagenomic analysis. The ranges of TPH concentrations in the grassland, abandoned well, working well soils were 1.16 × 102-3.50 × 102 mg/kg, 1.14 × 103-1.62 × 104 mg/kg, and 5.57 × 103-3.33 × 104 mg/kg, respectively. The highest concentration of n-alkanes and 16 PAHs were found in the working well soil of Shengli (SL) oilfield compared with those in Nanyang (NY) and Yanchang (YC) oilfields. The abandoned well soils showed a greater extent of petroleum biodegradation than the grassland and working well soils. Α-diversity indexes based on metagenomic taxonomy showed higher microbial diversity in grassland soils, whereas petroleum-degrading microbes Actinobacteria and Proteobacteria were more abundant in working and abandoned well soils. RDA demonstrated that low moisture content (MOI) in YC oilfield inhibited the accumulation of the petroleum-degrading microbes. Synergistic networks of functional genes and Spearman's correlation analysis showed that heavy petroleum contamination (over 2.10 × 104 mg/kg) negatively correlated with the abundance of the nitrogen fixation genes nifHK, however, in grassland soils, low petroleum content facilitated the accumulation of nitrogen fixation genes. A positive correlation was observed between the abundance of petroleum-degrading genes and denitrification genes (bphAa vs. nirD, todC vs. nirS, and nahB vs. nosZ), whereas a negative correlation was observed between alkB (alkane- degrading genes) and amo (ammonia oxidation), hao (nitrification). The ecotoxicity of petroleum contamination, coupled with petroleum hydrocarbons (PH) degradation competing with nitrifiers for ammonia inhibited ammonia oxidation and nitrification, whereas PH metabolism promoted the denitrification process. Moreover, positive correlations were observed between the abundance of amo gene and MOI, as well as between the abundance of the dissimilatory nitrate reduction gene nirA and clay content. Thus, improving the soil physicochemical properties is a promising approach for decreasing nitrogen loss and alleviating petroleum contamination in oilfield soils.

Preferential role of distinct phytochemicals in biosynthesis and antibacterial activity of silver nanoparticles.

Biosynthesis of silver nanoparticles (AgNPs) by plant extracts and its antibacterial utilization has attracted great attention due to the spontaneous reducing and capping capacities of phytochemicals. However, the preferential role and mechanisms of the functional phytochemicals from different plants on AgNPs synthesis, and its catalytic and antibacterial performance remain largely unknown. This study used three widespread arbor species, including Eriobotrya japonica (EJ), Cupressus funebris (CF) and Populus (PL), as the precursors and their leaf extracts as reducing and stabilizing agents for the biosynthesis of AgNPs. A total of 18 phytochemicals in leaf extracts were identified by ultra-high liquid-phase mass spectrometer. For EJ extracts, most kinds of flavonoids participated in the generation of AgNPs by a reduced content of 5∼10%, while for CF extracts, about 15∼40% of the polyphenols were consumed to reduce Ag+ to Ag0. Notably, the more stable and homogeneous spherical AgNPs with smaller size (≈38 nm) and high catalytic capacity on Methylene blue were obtained from EJ extracts rather than CF extracts, and no AgNPs were synthesized from PL extracts, indicating that flavonoids are superior than polyphenols to act as reducer and stabilizer in AgNPs biosynthesis. The antibacterial activities against Gram-positive (Staphylococcus aureus and Bacillus mycoides) and Gram-negative bacteria (Pseudomonas putida and Escherichia coli) were higher in EJ-AgNPs than that in CF-AgNPs, which confirmed the synergistic antibacterial effects of flavonoids combined with AgNPs in EJ-AgNPs. This study provides a significant reference on the biosynthesis of AgNPs with efficient antibacterial utilization underlying effect of abundant flavonoids in plant extracts.

New insight into the mechanisms of preferential encapsulation of metal(loid)s by wheat phytoliths under silicon nanoparticle amendment.

Silicon nanoparticles (SiNPs) have been widely used to immobilize toxic trace metal(loid)s (TTMs) in contaminated croplands. However, the effect and mechanisms of SiNP application on TTM transportation in response to phytolith formation and phytolith-encapsulated-TTM (PhytTTM) production in plants are unclear. This study demonstrates the promotion effect of SiNP amendment on phytolith development and explores the associated mechanisms of TTM encapsulation in wheat phytoliths grown on multi-TTM contaminated soil. The bioconcentration factors between organic tissues and phytoliths of As and Cr (> 1) were significantly higher than those of Cd, Pb, Zn and Cu, and about 10 % and 40 % of the total As and Cr that bioaccumulated in wheat organic tissues were encapsulated into the corresponding phytoliths under high-level SiNP treatment. These observations demonstrate that the potential interaction of plant silica with TTMs is highly variable among elements, with As and Cr being the two most strongly concentrated TTMs in the phytoliths of wheat treated with SiNPs. The qualitative and semi-quantitative analyses of the phytoliths extracted from wheat tissues suggest that the high pore space and surface area (≈ 200 m2 g-1) of phytolith particles could have contributed to the embedding of TTMs during silica gel polymerization and concentration to form PhytTTMs. The abundant SiO functional groups and high silicate-minerals in phytoliths are dominant chemical mechanisms for the preferential encapsulation of TTMs (i.e., As and Cr) by wheat phytoliths. Notably, the organic carbon and bioavailable Si of soils and the translocation of minerals from soil to plant aerial parts can impact TTM sequestration by phytoliths. Thus, this study has implications for the distribution or detoxification of TTMs in plants via preferential PhytTTM production and biogeochemical cycling of PhytTTMs in contaminated cropland following exogenous Si supplementation.

Nitrogen-doped biochar (N-doped BC) and iron/nitrogen co-doped biochar (Fe/N co-doped BC) for removal of refractory organic pollutants.

The presence of refractory organic pollutants (ROPs) in the ecosystem is a serious concern because of their impact on environmental constituents as well as their known or suspected ecotoxicity and adverse health effects. According to previous studies, carbonaceous materials, such as biochar (BC), have been widely used to remove pollutants from ecosystems owing to their desirable features, such as relative stability, tunable porosity, and abundant functionalities. Nitrogen (N)-doping and iron/nitrogen (Fe/N) co-doping can tailor BC properties and provide supplementary functional groups as well as extensive active sites on the N-doped and Fe/N co-doped BC surface, which is advantageous for interaction with and removal of ROPs. This review investigates the impact of N-doped and Fe/N co-doped BC on the removal of ROPs through adsorption, activation oxidation, and catalytic reduction due to the synergistic Fe, N, and BC features that modify the physicochemical properties, surface functional groups, and persistent free radicals of BC to aid in the degradation of ROPs. Owing to the attractive properties of N-doped and Fe/N co-doped BCs for the removal of ROPs, this review focuses and evaluates previous experimental investigations on the manufacturing (including precursors and influencing parameters during manufacturing) and characterizations of N-doped and Fe/N co-doped BCs. Additionally, the effective applications and mechanisms of N-doped and Fe/N co-doped BCs in adsorption, activation oxidation, and reductive remediation of ROPs are investigated herein. Moreover, the application of N-doped and Fe/N co-doped BC for progressive environmental remediation based on their effectiveness against co-pollutants, regeneration, stability, affordability, and future research prospects are discussed.

Fingerprint analysis reveals sources of petroleum hydrocarbons in soils of different geographical oilfields of China and its ecological assessment.

The distribution and characteristics of petroleum in three different geographic oilfields in China: Shengli Oilfield (SL), Nanyang Oilfield (NY), and Yanchang Oilfield (YC) were investigated. The average concentration of the total petroleum hydrocarbons (TPHs) conformed to be in the following law: SL Oilfield > NY Oilfield > YC Oilfield. Fingerprint analysis on the petroleum contamination level and source was conducted by the geochemical indices of n-alkanes and PAHs, such as low to high molecular weight (LMW/HMW) hydrocarbons, n-alkanes/pristine or phytane (C17/ Pr, C18/Ph), and ratio of anthracene/ (anthracene + phenanthrene) [Ant/(Ant + Phe)]. Soils adjacent to working well oils indicated new petroleum input with higher ratio of low to high molecular weight (LMW/HMW) hydrocarbons. The oil contamination occurred in the grassland soils might result of rainfall runoff. Petroleum source, petroleum combustion source, and biomass combustion were dominant PAHs origination of soils collected from oil exploitation area, petrochemical-related sites, farmland and grassland, respectively. The suggestive petroleum control strategies were proposed in each oilfield soils. Ecological potential risk of PAHs was assessed according to the toxic equivalent quantity (TEQ) of seven carcinogenic PAHs. The results showed that high, medium, and low ecological risk presented in petro-related area, grassland soils, and farmland soils, respectively. High ecological risk was persistent in abandoned oil well areas over abandoned time of 15 years, and basically stable after 5 years. This study can provide a critical insight to ecological risk management and source control of the petroleum contamination.

Enhanced degradation of petroleum hydrocarbons by immobilizing multiple bacteria on wheat bran biochar and its effect on greenhouse gas emission in saline-alkali soil.

In this study, an immobilization method for forming and keeping dominant petroleum degradation bacteria was successfully developed by immobilizing Pseudomonas, Acinetobacter, and Sphingobacterium genus bacteria on wheat bran biochar pyrolyzed at 300, 500, and 700 °C. The removal efficiency indicated that the highest TPHs (total petroleum hydrocarbons) removal rate of BC500-4 B (biochar pyrolyzed at 500 °C with four kinds of petroleum bacteria) was 58.31%, which was higher than that of BC500 (36.91%) and 4 B (43.98%) used alone. The soil properties revealed that the application of biochar increased the content of organic matter, available phosphorus, and available potassium, but decreased pH and ammonium nitrogen content in soil. Bacterial community analysis suggested that the formation of dominant degrading community represented by Acinetobacter played key roles in TPHs removal. The removal rate of alkanes was similar to that of TPHs. Besides, biochar and immobilized material can also mediate greenhouse gas emission while removing petroleum, biochar used alone and immobilized all could improve CO2 emission, but decrease N2O emission and had no significant impact on CH4 emission. Furthermore, it was the first time to found the addition of Acinetobacter genus bacteria can accelerate the process of forming a dominant degrading community in wheat bran biochar consortium. This study focused on controlling greenhouse gas emission which provides a wider application of combining biochar and bacteria in petroleum soil remediation.

Toxic mechanism on phenanthrene-induced cytotoxicity, oxidative stress and activity changes of superoxide dismutase and catalase in earthworm (Eisenia foetida): A combined molecular and cellular study.

Phenanthrene (PHE) is an important organic compound, which is widespread in the soil environment and exhibits potential threats to soil organisms. Toxic effects of PHE to earthworms have been extensively studied, but toxic mechanisms on PHE-induced cytotoxicity and oxidative stress at the molecular and cellular levels have not been reported yet. Therefore, we explored the cytotoxicity and oxidative stress caused by PHE in earthworm coelomocytes and the interaction mechanism between PHE and the major antioxidant enzymes SOD/CAT. It was shown that high-dose PHE exposure induced the intracellular reactive oxygen species (ROS) generation, mediated lipid peroxidation, reduced total antioxidant capacity (T-AOC) in coelomocytes, and triggered oxidative stress, thus resulted in a strong cytotoxicity at higher concentrations (0.6-1.0 mg/L). The intracellular SOD/CAT activity in cells after PHE exposure were congruent with that in molecular levels, which the activity of SOD enhanced and CAT inhibited. Spectroscopic studies showed the SOD/CAT protein skeleton and secondary structure, as well as the micro-environment of aromatic amino acids were changed after PHE binding. Molecular docking indicated PHE preferentially docked to the surface of SOD. However, the key residues Tyr 357, His 74, and Asn 147 for activity were in the binding pocket, indicating PHE more likely to dock to the active center of CAT. In addition, H-bonding and hydrophobic force were the primary driving force in the binding interaction between PHE and SOD/CAT. This study indicates that PHE can induce cytotoxicity and oxidative damage to coelomocytes and unearthes the potential effects of PHE on earthworms.

Effects of phosphorus modified nZVI-biochar composite on emission of greenhouse gases and changes of microbial community in soil.

The effect of modified biochar on the greenhouse gas emission in soil is not clear until now. In this study, biochar (BC) was modified by phosphoric acid (P) and further combined with nano-zero-valent iron (nZVI) to form nZVI-P-BC composite. The P modified biochar could significantly increase the available phosphorus in soil. The release of CO2 and N2O in soil was inhibited during the initial stage of the experiment, with inhibition becoming more obvious over time. On the contrary, CH4 and N2O emission in soil was enhanced by nZVI-P-BC composite. The proportion of Sphingomonas and Gemmatimonas were the most abundant bacterial species, which were related to the metabolism and transformation of nitrogen. The community structure of the fungus was also affected by nZVI-P-BC composite with Fusarium as the main species. PCoA analysis result suggested that bacterial community was more affected by the incubation time while fungal community was more related to the addition of different biochar and modified biochars.

High pyrolysis temperature biochar reduced the transport of petroleum degradation bacteria Corynebacterium variabile HRJ4 in porous media.

Biochar has been widely applied for the remediation of petroleum-contaminated soil. However, the effect of biochar on the transport of petroleum degradation bacteria has not been studied. A typical Gram-positive petroleum degradation bacteria-Corynebacterium variabile HRJ4 was used to study the effect of different biochars on bacterial transport and retention. Results indicated that the addition of biochar in sand was effective for reducing the transport of bacteria and poplar sawdust biochar (PSBC) had a stronger hinder effect than corn straw biochar (CSBC). The hindrance was more evident with pyrolysis temperature of biochar raised from 300°C to 600°C, which was attributed to the increase of specific surface area (309 times). The hindrance effect also enhanced with higher application rate of biochar. Furthermore, the reduction of HRJ4 transport was more obvious in higher (25 mmol/L) concentration of NaCl solution owing to electrostatic attraction enhancement. The adsorption of biochar to HRJ4 was defined to contribute to the hindrance of HRJ4 transport mainly. Combining the influence of feedstocks and pyrolysis temperature on HRJ4 transport, it suggested that specific surface area had the greatest effect on HRJ4 transport, and pore-filling, electrostatic force also contributed to HRJ4 retained in quartz sand column. At last, phenol transportation experiment indicated that the restriction of biochar on HRJ4 enhanced the phenol removal rate in the column. This study provides a theoretical basis for the interaction of biochar and bacteria, which is vital for the remediation of oil-contaminated soil and groundwater in the field.

Combination of rhamnolipid and biochar in assisting phytoremediation of petroleum hydrocarbon contaminated soil using Spartina anglica.

Biochar (BC) and rhamnolipid (RL) is used in bioremediation of petroleum hydrocarbons, however, the combined effect of BC and RL in phytoremediation has not been studied until now. In this paper, the phytoremediation of petroleum hydrocarbon-contaminated soil using novel plant Spartina anglica was enhanced by the combination of biochar (BC) and rhamnolipid (RL). Samples of petroleum-contaminated soil (10, 30 and 50 g/kg) were amended by BC, BC+ RL and rhamnolipid modified biochar (RMB), respectively. After 60 day's cultivation, the removal rate of total petroleum hydrocarbons (TPHs) for unplanted soil (UP), planted soil (P), planted soil with BC addition (P-BC), planted soil with BC and RL addition (P-BC + RL) and planted soil with addition of RMB (P-RMB) were 8.6%, 19.1%, 27.7%, 32.4% and 35.1% in soil with TPHs concentration of 30 g/kg, respectively. Compared with UP, the plantation of Spartina anglica significantly decreased the concentration of C8-14 and tricyclic PAHs. Furthermore, the application of BC and RMB alleviated the toxicity of petroleum hydrocarbons to Spartina anglica via improving plant growth with increasing plant height, root vitality and total chlorophyll content. High-throughput sequencing result indicated that rhizosphere microbial community of Spartina anglica was regulated by the application of BC and RMB, with increase of bacteria and plant mycorrhizal symbiotic fungus in biochar and RMB amended soil.

Exposure to Al2O3 nanoparticles facilitates conjugative transfer of antibiotic resistance genes from Escherichia coli to Streptomyces.

The spread of antibiotic resistance genes (ARGs) has become a global environmental issue; it has been found that nanoparticles (NPs) can promote the transfer of ARGs between bacteria. However, it remains unclear whether NPs can affect this kind of conjugation in Streptomyces, which mainly conjugate with other bacteria via spores. In the present study, we demonstrated that Al2O3 NPs significantly promote the conjugative transfer of ARGs from Escherichia coli (E. coli) ET12567 to Streptomyces coelicolor (S. coelicolor) M145 without the use of heat shock method. The number of transconjugants induced by Al2O3 particles was associated with the size and concentration of Al2O3 particles, exposure time, and the ratio of E. coli and spores. When nanoparticle size was 30 nm at a concentration of 10 mg/L, the conjugation efficiency reached a peak value of 182 cfu/108 spores, which was more than 60-fold higher than that of the control. Compared with nanomaterials, bulk particles exhibited no significant effect on conjugation efficiency. We also explored the mechanisms by which NPs promote conjugative transfer. After the addition of NPs, the intracellular ROS content increased and the expression of the classical porin gene ompC was stimulated. In addition, ROS enhanced the mRNA expression levels of conjugative genes by inhibiting global regulation genes. Meanwhile, expression of the conjugation-related gene intA was also stimulated, ultimately increasing the number of transconjugants. Our results indicated that Al2O3 NPs significantly promoted the conjugative transfer of ARGs from bacteria to spores and aggravated the diffusion of resistance genes in the environment.

Effect of rhamnolipids on enhanced anaerobic degradation of petroleum hydrocarbons in nitrate and sulfate sediments.

Anaerobic degradation of petroleum hydrocarbons (PH) is an important process in contaminated environment. The application of rhamnolipids in anaerobic degradation of PH was not extensively studied and inconclusive. This study explored the combined effect of rhamnolipids and electron acceptors on the anaerobic degradation process of total petroleum hydrocarbons (TPH) in sediment from an oil field. The results indicated that rhamnolipids decreased the surface tension of the medium and increased the desorption of TPH from the sediment. After 10-wk culture, the maximum degradation rate of TPH in nitrate and sulfate condition was found to be 32.2% and 24.0%, respectively, with rhamnolipids concentration of 150 mg/L. The addition of 45 and 150 mg/L rhamnolipids increased the degradation rate of TPH but the promotion effect was weakened in the treatment with 450 mg/L rhamnolipids. The copy number of two degradation genes (1-methylalkyl) succinate synthase gene (masD) and 6-oxocyclohex-1-ene-1-carbonyl-CoA hydrolase gene (bamA) increased with incubation time and showed higher copy numbers in treatments with 45 and 150 mg/L rhamnolipids. In the first week, with the increase of rhamnolipids concentration, the copy number of 16S rDNA increased rapidly and the concentration of electron receptors decreased correspondingly. Moreover, no nitrate was detected in treatments of nitrate with 450 mg/L rhamnolipids after the first week. Microbial community structure analysis result showed that Thiobacillus was the dominant bacteria in all treatments with nitrate as electron acceptor and its proportion gradually decreased with the increase of rhamnolipids concentration. The addition of rhamnolipids changed the subdominant bacteria in the treatments with nitrate as electron acceptor. Methanothrix was the dominant archaea in all treatments with rhamnolipids content of lower than 45 mg/L. When the rhamnolipids concentration increased, the dominant archaea changed to Methanogenium or Methanobacterium. In conclusion, suitable concentrations of rhamnolipids could promote the anaerobic degradation of PH in the sediment.

Interactions between electrokinetics and rhizoremediation on the remediation of crude oil-contaminated soil.

An electrokinetics (EK)-enhanced phytoremediation system with ryegrass was constructed to remediate crude oil-polluted soil. The four treatments employed in this study included (1) without EK or ryegrass (CK-NR), (2) EK only (EK-NR), (3) ryegrass only (CK-R), and (4) EK and ryegrass (EK-R). After 30d of ryegrass growth, EK at 1.0 V·cm-1 with polarity reversal (PR-EK) was supplied for another 30 d. The electric current was recorded during remediation. The pH, electrical conductivity, total petroleum hydrocarbon content (TPH), 16S rDNA, functional genes of AlkB, Nah, and Phe, DGGE, and dehydrogenase activity in soil were measured. The physical-chemical indexes of the plant included the length, dry mass, and chlorophyll contents of the ryegrass. Results showed that EK-R removed 18.53 ±â€¯0.53% of TPH, which was higher than that of other treatments (13.34-14.31%). Meanwhile, the values of 16S rDNA, AlkB, Nah, Phe, and dehydrogenase activity in the bulk soil of EK-R all increased. Further clustering analysis with numbers of genes and DGGE demonstrated that EK-R was similar to the ryegrass rhizosphere soils in both EK-R and CK-R, while the EK treatment of EK-NR was similar to that of CK-NR without EK and ryegrass. These results indicate that the PR-EK treatment used in this experiment successfully enlarged the existing scale of the rhizosphere microorganisms, improved microbial activity and enhanced degradation of TPH.

Vertical response of microbial community and degrading genes to petroleum hydrocarbon contamination in saline alkaline soil.

A column microcosm was conducted by amending crude oil into Dagang Oilfield soil to simulate the bioremediation process. The dynamic change of microbial communities and metabolic genes in vertical depth soil from 0 to 80 cm were characterized to evaluate the petroleum degradation potential of indigenous microorganism. The influence of environmental variables on the microbial responds to petroleum contamination were analyzed. Degradation extent of 42.45% of n-alkanes (C8-C40) and 34.61% of 16ΣPAH were reached after 22 weeks. Relative abundance of alkB, nah, and phe gene showed about 10-fold increment in different depth of soil layers. Result of HTS profiles demonstrated that Pseudomonas, Marinobacter and Lactococcus were the major petroleum-degrading bacteria in 0-30 and 30-60 cm depth of soils. Fusarium and Aspergillus were the dominant oil-degrading fungi in the 0-60 cm depth of soils. In 60-80 cm deep soil, anaerobic bacteria such as Bacteroidetes, Lactococcus, and Alcanivorax played important roles in petroleum degradation. Redundancy analysis (RDA) and correlation analysis demonstrated that petroleum hydrocarbons (PHs) as well as soil salinity, clay content, and anaerobic conditions were the dominant effect factors on microbial community compositions in 0-30, 30-60, and 60-80 cm depth of soils, respectively.

Influence of graphene oxide and biochar on anaerobic degradation of petroleum hydrocarbons.

The anaerobic degradation of petroleum is an important process in natural environments. So far, few studies have considered the response of the microbial community to nanomaterials during this process. This study explored the potential effects of graphene oxide and biochar on the anaerobic degradation of petroleum hydrocarbons in long-term experiments. Cyclic voltammetry and electrochemical impedance spectroscopy indicated that the addition of carbon-based materials promoted the electrochemical activity of anaerobic cultures that degrade petroleum hydrocarbons. The maximum degradation rates for benzene, toluene, ethylbenzene, and xylene (BTEXs) in the cultures incubated for 10 weeks with graphene oxide (0.02 mg/L) and biochar (20 mg/L) were 76.5% and 77.6%, respectively. The maximum degradation rates of n-alkanes in the cultures incubated for 10 weeks with graphene oxide (2 mg/L) and biochar (100 mg/L) were 70.0% and 77.8%, respectively. The 16S rDNA copy numbers in the treatments with 0.02 mg/L graphene oxide and 20 mg/L biochar were significantly higher than others during the process (P < 0.05). In the 2nd week, the maximum copy numbers of the masD and bamA genes in the treatments with biochar were 349 copies/mL (20 mg/L) and 422 copies/mL (20 mg/L), respectively, and in the treatments with graphene oxide were 289 copies/mL (0 mg/L) and 366 copies/mL (0.02 mg/L). The contents of carbon-based materials had slight effects on the microbial community structure, whereas the culture time had obvious effects. Paracoccus denitrificans, Pseudomonas aeruginosa, and Hydrogenophaga caeni were the dominant microorganisms in the culture systems under all treatments.

Aerobic degradation of crude oil by microorganisms in soils from four geographic regions of China.

A microcosm experiment was conducted for 112 d by spiking petroleum hydrocarbons into soils from four regions of China. Molecular analyses of soils from microcosms revealed changes in taxonomic diversity and oil catabolic genes of microbial communities. Degradation of total petroleum hydrocarbons (TPHs) in Sand from the Bohai Sea (SS) and Northeast China (NE) exhibited greater microbial mineralization than those of the Dagang Oilfield (DG) and Xiamen (XM). High-throughput sequencing and denaturing gradient gel electrophoresis (DGGE) profiles demonstrated an obvious reconstruction of the bacterial community in all soils. The dominant phylum of the XM with clay soil texture was Firmicutes instead of Proteobacteria in others (DG, SS, and NE) with silty or sandy soil texture. Abundances of alkane monooxygenase gene AlkB increased by 10- to 1000-fold, relative to initial values, and were positively correlated with rates of degradation of TPHs and n-alkanes C13-C30. Abundances of naphthalene dioxygenase gene Nah were positively correlated with degradation of naphthalene and total tricyclic PAHs. Redundancy analysis (RDA) showed that abiotic process derived from geographical heterogeneity was the primary effect on bioremediation of soils contaminated with oil. The optimization of abiotic and biotic factors should be the focus of future bioremediation of oil contaminated soil.

Degradation of n-alkanes and PAHs from the heavy crude oil using salt-tolerant bacterial consortia and analysis of their catabolic genes.

In the present study, salt-tolerant strains, Dietzia sp. HRJ2, Corynebacterium variabile HRJ4, Dietzia cinnamea HRJ5 and Bacillus tequilensis HRJ6 were isolated from the Dagang oil field, China. These strains degraded n-alkanes and polycyclic aromatic hydrocarbons (PAHs) aerobically from heavy crude oil (HCO) in an experiment at 37 °C and 140 rpm. The GC/MS investigation for degradation of different chain lengths of n-alkanes (C8-C40) by individual strains showed the highest degradation of C8-C19 (HRJ5), C20-C30 (HRJ4) and C31-C40 (HRJ5), respectively. Moreover, degradation of 16 PAHs with individual strains demonstrated that the bicyclic and pentacyclic aromatic hydrocarbons (AHs) were mostly degraded by HRJ5, tricyclic and tetracyclic AHs by HRJ6 and hexacyclic AHs by HRJ2. However, the highest degradation of total petroleum hydrocarbons (TPHs), total saturated hydrocarbons (TSH), total aromatic hydrocarbons (TAH), n-alkanes (C8-C40) and 16 PAHs was achieved by a four-membered consortium (HRJ2 + 4 + 5 + 6) within 12 days, with the predominance of HRJ4 and HRJ6 strains which was confirmed by denaturing gradient gel electrophoresis. The abundance of alkB and nah genes responsible for catabolism of n-alkanes and PAHs was quantified using the qPCR. Maximum copy numbers of genes were observed in HRJ2 + 4 + 5 + 6 consortium (gene copies l-1) 2.53 × 104 (alkB) and 3.47 × 103 (nah) at 12 days, which corresponded to higher degradation rates of petroleum hydrocarbons. The superoxide dismutase (SOD) (total SOD (T-SOD), Cu2+Zn2+-SOD), catalase (CAT) and ascorbate peroxidase (APX) activities in Allium sativum and Triticum aestivum were lower in the HRJ2 + 4 + 5 + 6-treated HCO as compared to the plantlets exposed directly to HCO. The present results revealed the effective degradation of HCO-contaminated saline medium using the microbial consortium having greater metabolic diversity.

A novel bioremediation strategy for petroleum hydrocarbon pollutants using salt tolerant Corynebacterium variabile HRJ4 and biochar.

The present work aimed to develop a novel strategy to bioremediate the petroleum hydrocarbon contaminants in the environment. Salt tolerant bacterium was isolated from Dagang oilfield, China and identified as Corynebacterium variabile HRJ4 based on 16S rRNA gene sequence analysis. The bacterium had a high salt tolerant capability and biochar was developed as carrier for the bacterium. The bacteria with biochar were most effective in degradation of n-alkanes (C16, C18, C19, C26, C28) and polycyclic aromatic hydrocarbons (NAP, PYR) mixture. The result demonstrated that immobilization of C. variabile HRJ4 with biochar showed higher degradation of total petroleum hydrocarbons (THPs) up to 78.9% after 7-day of incubation as compared to the free leaving bacteria. The approach of this study will be helpful in clean-up of petroleum-contamination in the environments through bioremediation process using eco-friendly and cost effective materials like biochar.

Distribution of petroleum degrading genes and factor analysis of petroleum contaminated soil from the Dagang Oilfield, China.

Genes that encode for enzymes that can degrade petroleum hydrocarbons (PHs) are critical for the ability of microorganisms to bioremediate soils contaminated with PHs. Distributions of two petroleum-degrading genes AlkB and Nah in soils collected from three zones of the Dagang Oilfield, Tianjin, China were investigated. Numbers of copies of AlkB ranged between 9.1 × 10(5) and 1.9 × 10(7) copies/g dry mass (dm) soil, and were positively correlated with total concentrations of PHs (TPH) (R(2) = 0.573, p = 0.032) and alkanes (C33 ~ C40) (R(2) = 0.914, p < 0.01). The Nah gene was distributed relatively evenly among sampling zones, ranging between 1.9 × 10(7) and 1.1 × 10(8) copies/g dm soil, and was negatively correlated with concentrations of total aromatic hydrocarbons (TAH) (R(2) = -0.567, p = 0.035) and ∑16 PAHs (R(2) = -0.599, p = 0.023). Results of a factor analysis showed that individual samples of soils were not ordinated as a function of the zones.

Eco-toxicity of petroleum hydrocarbon contaminated soil.

Total petroleum hydrocarbons (TPH) contaminated soil samples were collected from Shengli Oilfield of China. Toxicity analysis was carried out based on earthworm acute toxicity, plant growth experiment and luminescent bacteria test. The soil was contaminated by-petroleum hydrogcarbons with TPH concentration of 10.57%. With lethal and sub-lethal rate as endpoint, earthworm test showed that the LD50 (lethal dose 50%) values in 4 and 7 days were 1.45% and 1.37% respectively, and the inhibition rate of earthworm body weight increased with higher oil concentration. TPH pollution in the soil inhibited seed germination in both wheat and maize experiment when the concentration of petroleum was higher than 0.1%. The EC50 (effective concentration 50%) for germination is 3.04% and 2.86% in maize and wheat, respectively. While lower value of EC50 for root elongation was to be 1.11% and 1.64% in maize and wheat, respectively, suggesting higher sensitivity of root elongation on petroleum contamination in the soil. The EC50 value in luminescent bacteria test was 0.47% for petroleum in the contaminated soil. From the experiment result, it was concluded that TPH content of 1.5% is considered to be a critical value for plant growth and living of earthworm and 0.5% will affect the activity of luminescent bacteria.