Synthesis of iron-manganese bimetallic materials supported by activated carbon and application of activated persulfate in the degradation of soil contaminated by crude oil.

Heterogeneous catalytic processes based on zero-valent iron (ZVI) have been developed to treat soil and wastewater pollutants. However, the agglomeration of ZVI reduces its ability to activate persulfate (PS). In this study, a new Fe-Mn@AC activated material was prepared to activated PS to treat oil-contaminated soil, and using the microscopic characterization of Fe-Mn@AC materials, the electron transfer mode during the Fe-Mn@AC activation of PS was clarified. Firstly, the petroluem degradation rate was optimized. When the PS addition amount was 8%, Fe-Mn@AC addition amount was 3% and the water to soil ratio was 3:1, the petroluem degradation rate in the soil reached to the maximum of 85.69% after 96 h of reaction. Then it was illustrated that sulfate and hydroxyl radicals played major roles in crude oil degradation, while singlet oxygen contributed slightly. Finally, the indigenous microbial community structures remaining after restoring the Fe-Mn@AC/PS systems were analyzed. The proportion of petroleum degrading bacteria in soil increased by 23% after oxidation by Fe-Mn@AC/PS system. Similarly, the germination rate of wheat seeds revealed that soil toxicity was greatly reduced after applying the Fe-Mn@AC/PS system. After the treatment with Fe-Mn@AC/PS system, the germination rate, root length and bud length of wheat seed were increased by 54.05%, 7.98 mm and 6.84 mm, respectively, compared with the polluted soil group. These results showed that the advanced oxidation system of Fe-Mn@AC activates PS and can be used in crude oil-contaminated soil remediation.

Response of petroleum-contaminated soil to chemical oxidation combined with biostimulation.

In this study, a microcosm experiment was conducted to investigate the effects of Na2S2O8 preoxidation combined with biostimulation on petroleum-contaminated soil remediation. The response of microbial community during this process was explored using BIOLOG ECO microplate carbon utilization method and 16 s rDNA high-throughput sequencing. The results showed that use of 10 mg/g Na2S2O8 removed 19.8 % of the petroleum hydrocarbons, reduced soil biotoxicity and did not affect soil microbial activity compared to other concentrations. Therefore, sodium persulfate of ca. 10 mg/g was used to oxidize petroleum in soil before the biostimulation experiment with organic and inorganic fertilizers. Our finding showed that the content of total petroleum hydrocarbons (TPHs) in soil was reduced by 43.3 % in inorganic fertilizer treatment after 60 days. The results of BIOLOG ECO microplate carbon utilization analysis and 16 S rDNA high-throughput sequencing further confirmed that biostimulation quickly restored the microbial activities in oxidant treated soil. The main marker bacteria in chemical oxidation combined with biostimulation remediation were Arthrobacter and Paenarthrobacter, and their relative abundances were both significantly negatively correlated with the content of petroleum hydrocarbons in soil.

Surface Characteristics of Activated Carbon Sorbents Obtained from Biomass for Cleaning Oil-Contaminated Soils.

This study explores the sorption capacity and field application of activated carbons (ACs) derived from plant residues for the remediation of oil-contaminated soils. ACs were prepared from rice husks, reed stalks, pine sawdust and wheat straw using two-stage pyrolysis and chemical activation with potassium hydroxide. The structural and physicochemical properties of these ACs were analyzed using BET surface area measurements, SEM analysis, Raman spectroscopy and FTIR spectroscopy. Sorption experiments at room temperature demonstrated that AC from rice husks (OSL) exhibited the highest sorption capacities for gasoline, kerosene and diesel fuel, with values of 9.3 g/g, 9.0 g/g and 10.1 g/g, respectively. These results are attributed to the well-developed microporous and mesoporous structures of OSL, as confirmed by SEM images and a BET surface area of 2790 m2/g. Field tests conducted at the "Zhanatalap" oil deposit showed that the ACs effectively reduced the oil content in contaminated soils from 79.2 g/kg to as low as 2.6 g/kg, achieving a purification degree of up to 67% within 16 days. This study highlights the critical role of structural properties, such as porosity and graphitization degree, in enhancing the sorption efficiency of ACs.

Enhancing biodegradation of vegetable oil-contaminated soil with soybean texturized waste, spent mushroom substrate, and stabilized poultry litter in microcosm systems.

Industrial activities contribute to environmental pollution, particularly through unregulated effluent discharges, causing adverse effects on ecosystems. Vegetable oils, as insoluble substances, exacerbate this pollution, forming impermeable films and affecting the oxygen transfer, leading to serious habitat disruption. Organic wastes, such as soybean texturized waste, spent mushroom substrate, and stabilized poultry litter, were assessed for their efficacy in enhancing the degradation of vegetable oil in contaminated soil. For this purpose, contaminated soil was amended with each of the wastes (10% w/w) using microcosm systems, which were monitored physico-chemically, microbiologically and toxicologically. Results indicate that the wastes promoted significant oil degradation, achieving 83.1, 90.7, and 86.2% removal for soybean texturized waste, spent mushroom substrate, and stabilized poultry litter, respectively, within a 90-day period. Additionally, they positively influenced soil microbial activity, as evidenced by increased levels of culturable microorganisms and hydrolytic microbial activity. While bioassays indicated no phytotoxicity in most cases, soybean texturized waste exhibited inhibitory effects on seed germination and root elongation of Lactuca sativa. This study significantly enhances our comprehension of remediation techniques for sites tainted with vegetable oils, highlighting the critical role of organic waste as eco-friendly agents in soil restoration. Emphasizing the practical implications of these findings is imperative to underscore the relevance and urgency of addressing vegetable oil contamination in soil. Moving forward, tailored strategies considering both contaminant characteristics and soil ecosystem traits are vital for ensuring effective and sustainable soil remediation.

Enhanced crude oil degradation by remodeling of crude oil-contaminated soil microbial community structure using sodium alginate/graphene oxide/Bacillus C5 immobilized pellets.

Bioaugmentation (BA) of oil-contaminated soil by immobilized microorganisms is considered to be a promising technology. However, available high-efficiency microbial agents remain very limited. Therefore, we prepared a SA/GO/C5 immobilized gel pellets by embedding the highly efficient crude oil degrading bacteria Bacillus C5 in the SA/GO composite material. The optimum preparation conditions of SA/GO/C5 immobilized gel pellets were: SA 3.0%, GO 25.0 µg/mL, embedding amount of C5 6%, water bath temperature of 50°C, CaCl2 solution concentration 3% and cross-linking time 20 h. BA experiments were carried out on crude oil contaminated soil to explore the removal effect of SA/GO/C5 immobilized pellets. The results showed that the SA/GO/C5 pellets exhibited excellent mechanical strength and specific surface area, which facilitated the attachment and growth of the Bacillus C5. Compared with free bacteria C5, the addition of SA/GO/C5 significantly promoted the removal of crude oil in soil, reaching 64.92% after 30 d, which was 2.1 times the removal rate of C5. The addition of SA/GO/C5 promoted the abundance of soil exogenous Bacillus C5 and indigenous crude oil degrading bacteria Alcanivorax and Marinobacter. In addition, the enrichment of hydrocarbon degradation-related functional abundance was predicted by PICRUSt2 in the SA/GO/C5 treatment group. This study demonstrated that SA/GO/C5 is an effective method for remediating crude oil-contaminated soil, providing a basis and option for immobilized microorganisms bioaugmentation to remediate organic contaminated soil.

Concentrations and health risk appraisal of heavy metals and volatile organic compounds in soils of automobile mechanic villages in Ogun State, Nigeria.

This report presents the findings of the concentrations, distributions and health risks assessment of heavy metals (HMs) and volatile organic compounds (VOCs) in topsoils of two typical automobile mechanic villages (MVs) situated within Ogun State, Nigeria. One of the MVs is located in basement complex terrain (Abeokuta), while the second is in the sedimentary formation (Sagamu). Ten composite samples were collected at depth of 0-30 cm with the aid of soil auger from spent oil-contaminated spots within the two MVs. The chemical parameters of interest were Pb, Cd, benzene, ethylbenzene, toluene, total petroleum hydrocarbon (TPH) as well as oil and grease (O&G). In addition, soil pH, cation exchange capacity (CEC), electrical conductivity (EC) and particle size distribution were also evaluated in order to find out their impacts on assessed soil pollutants. Results revealed that the soils in both MVs are of sandy loam texture, slight acidic to neutral pH, mean CEC < 15 cmol/kg and mean EC > 100 µS/cm. The mean concentration of each of analyzed HMs and VOCs in soils from the two MVs was < 5 mg/kg, while the mean values of TPH and O&G content were > 50 mg/kg. The mean Cd values in soils of both MVs were higher than the national soil screening level of 0.8 mg/kg, but lower than the Canadian and Italian guidelines. There is no significant correlation between each of HMs/VOCs and any of assessed soil physicochemical variables. The non-cancer risk expressed in terms of hazard index (HI) was > 1 via oral ingestion route for adults and children at the two MVs, indicating adverse non-carcinogenic health risk. The HI > 1 value was obtained for adults only through the dermal absorption pathway in Abeokuta MV. However, HI values for the two age groups at the two MVs via inhalation route were < 1, indicating no likelihood of any non-carcinogenic effects via the breathing exposure. The potential of non-cancer risk via oral ingestion route in both MVs was derived from the contributive ratios of HMs and VOCs in the order: Cd > benzene > Pb > toluene. The carcinogenic risk (CR) values due to ingested Cd, benzene and Pb for both age groups at the two MVs exceed the safe limit range of 10-6 to 10-4. Cadmium, benzene and lead made considerable contributions to the estimation of CR through dermal exposure for adults only in Abeokuta MV. The CR values via inhalation pathway for adults and children in both MVs were within the threshold range. Artisans and children should circumvent accidental ingestion of contaminated soils in addition to wearing of protective clothes during routine vehicle maintenance activities.

Kaistella soli sp. nov., isolated from oil-contaminated experimental soil.

A light yellow-coloured, non-motile, aerobic, Gram-stain-negative, and rod-shaped bacterial strain DKR-2T was isolated from oil-contaminated experimental soil. The strain was catalase and oxidase positive, and grew at 0-1.5% (w/v) NaCl concentration, at temperature 10-35 °C, and at pH 6.0-9.5. The phylogenetic analysis suggested that the strain DKR-2T was affiliated to the genus Kaistella, with the closest species being Kaistella haifensis DSM 19056T (97.6% 16S rRNA gene sequence similarity). The principle fatty acids were iso-C15:0, summed feature 9 (iso-C17:1 ω9c and/or C16:0 10-methyl), and antiso-C15:0. The sole menaquinone was MK-6 and major polar lipid was phosphatidylethanolamin. The DNA G+C content was 39.5%. The dDDH (in silico DNA-DNA hybridization) and ANI (average nucleotide identity) values between strain DKR-2T and K. haifensis DSM 19056T were 22.4% and 79.3%, respectively. In addition, both dDDH and ANI values between strain DKR-2T and other phylogenetically related neighbours were < 25.0% and < 77.0%, respectively. In overall, the polyphasic taxonomic data presented in this study clearly indicated that strain DKR-2T represents a novel species in the genus Kaistella, for which the name Kaistella soli sp. nov. is proposed. The type strain is DKR-2T (=KACC 22070T=NBRC 114725T).

Cellulomonas fulva sp. nov., isolated from oil-contaminated soil.

A yellow-coloured, Gram-stain-positive, motile, aerobic and rod-shaped bacteria, designated DKR-3T, was isolated from oil-contaminated experimental soil. Strain DKR-3T could grow at pH 5.0-10.5 (optimum, pH 7.0-8.5), at 10-40 °C (optimum, 25-32 °C) and tolerated 3.5 % of NaCl. Phylogenetic analyses based on its 16S rRNA gene sequence indicated that strain DKR-3T formed a lineage within the family Cellulomonadaceae and was clustered with members of the genus Cellulomonas. Strain DKR-3T had highest 16S rRNA gene sequence similarities to Cellulomonas gelida DSM 20111T (98.3 %), Cellulomonas persica JCM 18111T (98.2 %) and Cellulomonas uda DSM 20107T (97.8 %). The predominant respiratory quinone was tetrahydrogenated menaquinone with nine isoprene units [MK-9(H4)]. The principal cellular fatty acids were anteiso-C15 : 0, C16 : 0 and anteiso-C17 : 0. The major polar lipids were diphosphatidylglycerol and phosphatidylglycerol. The cell-wall diamino acid was l-ornithine whereas rhamnose and glucose were the cell-wall sugars. The DNA G+C content was 74.2mol %. The genome of strain DKR-3T was 3.74 Mb and contained three putative biosynthetic gene clusters. The average nucleotide identity and digital DNA-DNA hybridization relatedness values between strain DKR-3T and its phylogenetically related members were below the species threshold values. Based on a polyphasic study, strain DKR-3T represents a novel species belonging to the genus Cellulomonas, for which the name Cellulomonas fulva sp. nov. is proposed. The type strain is DKR-3T (=KACC 22071T=NBRC 114730T).

In situ microbial and phytoremediation of crude oil contaminated soil by Cynodon sp.

Crude oil contamination of land and water leads to their abandonment after heavy oil recovery processes. Analogous to bioremediation, phytoremediation has provided an efficient solution towards land reclamation through enhancement of flora. The present work manifests significance of phytoremediation via reclamation of crude oil contaminated soil collected from Kalol, India. The collected soil was analyzed for pH, oxidation-reduction potential, electrical conductivity (EC), bulk density, particle size, moisture. The experimental work consists three batch units; pot A, pot B and pot C with crude oil contaminated soil, fresh soil and control respectively. While observing plant growth for 120 days, Total Petroleum Hydrocarbon (TPH) was measured at determined intervals for estimation of percentage degradation. After 90 days of pot observation, contaminated soil was inoculated with rhizospheric bacterial inoculum developed from pot A which forms new batch for microbial-remediation as an additional scope to this work. Gas chromatography mass spectroscopy (GC-MS-MS) was carried out for determination of naphthalene contamination. Crude oil degradation in pot A was estimated as 82.16% followed with the affirmation given by degradation kinetics whereas, 60.68% and 36.75% degradation was observed in pot C-control and new batch respectively. Cynodon sp. grown in pot A was confirmed by identification as reported.

Efficient removal of heavily oil-contaminated soil using a combination of fenton pre-oxidation with biostimulated iron and bioremediation.

Crude oil contamination severely deteriorates soils quality. Bioremediation utilizing soil indigenous organisms could be employed to decompose petroleum hydrocarbons thanks to its low cost and minor environmental disturbance. However, slow kinetics limit the successful application of this biotechnique. Pretreating oil-contaminated soils with Fenton pre-oxidation could accelerate the subsequent bioremediation process. This study was to explore the mechanisms behind the rapid propagation of indigenous petroleum-degrading bacteria (IPDB) and the efficient degradation of total petroleum hydrocarbons (TPH) in soil after Fenton pre-oxidation with biostimulated iron. Biostimulated iron and non-biostimulated iron were used in the experiments, where Fenton pre-oxidation was combined with the bioremediation of oil-contaminated soil (TPH = 13221 mg/kg). Although the amount of Fenton pre-oxidized TPH (3331-3775 mg/kg) was similar with biostimulated and non-biostimulated irons, the biodegradation of TPH after Fenton pre-oxidation with biostimulated iron (5840 mg/kg) was much higher than that with non-biostimulated iron (3034-4034 mg/kg). Moreover, abundant nutrients and a high population of residual IPDB were found after Fenton pre-oxidation with biostimulated iron, which benefited stable consumption of NH3-N and dissolved organic carbon (DOC) by IPDB during the subsequent bioremediation. However, Fenton pre-oxidation with non-biostimulated iron either resulted in greater damage to IPDB or produced fewer nutrients, thereby failing to ensure the continuous propagation of IPDB during the subsequent bioremediation. Therefore, we propose that Fenton pre-oxidation with biostimulated iron should be applied to heavily oil-contaminated soils prior to bioremediation.

Fast biodegradation of long-alkanes by enhancing bacteria performance rate by per-oxidation.

The long-alkanes biodegradation rate was generally found slow during widely used pre-oxidation combined with biodegradation for oil contamination treatment, resulting in long and unsustainable removal. In this study, different chitosan content was used to produce iron catalysts for pre-oxidation, and nutrients were added for the long-alkanes biodegradation experiment. Mechanism of Fenton pre-oxidation and improvement in the biodegradation rate of long-alkanes were studied by analyzing the change in organic matter and bacterial community structure, the amount and activity of bacteria in the biological stage, and the degradation amount long-alkanes hydrocarbon before and after pre-oxidation. Results showed that the destruction of bacteria greatly reduced when hydroxyl radical intensity decreased to 4.40 a.u.. Also, the proportion of humic acid-like was high (40.88%), and the community structure was slightly changed with the pre-oxidation for the fast biodegradation (FB) group. In the subsequent biodegradation, it was found that the degradation rate of each long-alkanes in the FB group increased significantly (C30: 4.18-8.32 mg/(kg·d)) with the increase of the degradation of long-alkanes (10-50%). Further studies showed that the high nutrient dynamics (6.05 mg/(kg·d)) of the FB group resulted in high bacteria performance rate (0.53 mol CO2 × log CFU/(104 g2 d)), which further accelerated the substrate transformation(41%). Therefore, the biodegradation rate of long-alkanes was increased (43.8 mg/(kg·d)) with the removal rate of long-alkanes of 76%. The half-life of long-alkanes for the FB group (64 d) was 33 d shorter than the slow biodegradation group (99 d). These results exhibited that pre-oxidation regulation can shorten the bioremediation cycle by improving the biodegradation rate of long-alkanes. This research has good engineering application value.

Noviherbaspirillum pedocola sp. nov., isolated from oil-contaminated experimental soil.

An orange-coloured, rod-shaped, and aerobic bacterial strain DKR-6 T was isolated from oil-contaminated experimental soil. The strain was Gram-stain-negative, catalase and oxidase positive, and grew at temperature 10-42 °C, at pH 5.5-9.5, and at 0-3.0% (w/v) NaCl concentration. The phylogenetic analysis and 16S rRNA gene sequence analysis suggested that the strain DKR-6 T was affiliated to the genus Noviherbaspirillum, with the closest species being Noviherbaspirillum massiliense JC206T (96.3% sequence similarity). The chemotaxonomic profiles revealed the presence of phosphatidylethanolamine, phosphatidylglycerol, diphosphatidylglycerol, and phosphatidylcholine as the principal polar lipids; C16:0, C17:0 cyclo, summed feature 3 (C16:1ω7c and/or C16: 1ω6c), and summed feature 8 (C18:1ω7c/or C18:1ω6c) as the main fatty acids; and Q-8 as a sole ubiquinone. The DNA G + C content was 61.6%. The polyphasic taxonomic features illustrated in this study clearly implied that strain DKR-6 T represents a novel species in the genus Noviherbaspirillum, for which the name Noviherbaspirillum pedocola sp. nov. is proposed with the type strain DKR-6 T (= KACC 22074 T = NBRC 114727 T).

Novosphingobium olei sp. nov., with the ability to degrade diesel oil, isolated from oil-contaminated soil and proposal to reclassify Novosphingobium stygium as a later heterotypic synonym of Novosphingobium aromaticivorans.

Two yellow-pigmented, non-motile, Gram-stain-negative, and rod-shaped bacteria, designated TW-4T and TNP-2 were obtained from oil-contaminated soil. Both strains degrade diesel oil, hydrolyse aesculin, DNA, Tween 40 and Tween 60. A phylogenetic analysis based on its 16S rRNA gene sequence revealed that strain TW-4T formed a lineage within the family Erythrobacteraceae and clustered as members of the genus Novosphingobium. The closest members of strain TW-4T were Novosphingobium subterraneum DSM 12447T (97.9 %, sequence similarity), Novosphingobium lubricantis KSS165-70T (97.8 %), Novosphingobium taihuense T3-B9T (97.8 %), Novosphingobium aromaticivorans DSM 12444T (97.7 %), Novosphingobium flavum UCT-28T (97.7 %), and Novosphingobium bradum STM-24T (97.6 %). The sequence similarity for other members was ≤97.6 %. The genome of strain TW-4T was 4 683 467 bp long with 44 scaffolds and 4280 protein-coding genes. The sole respiratory quinone was Q-10. The major cellular fatty acids were summed feature 8 (C18 : 1 ω7c and/or C18 : 1 ω6c), summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), C16 : 0 and C14 : 0 2-OH. The major polar lipids were phosphatidylethanolamine (PE), phosphatidylglycerol (PG), diphosphatidylglycerol (DPG), phosphatidylcholine (PC), phosphatidyl-n-methylethanolamine (PME) and sphingoglycolipid (SGL). The DNA G+C content of the type strain was 65.0 %. The average nucleotide identity (ANIu) and in silico DNA-DNA hybridization (dDDH) relatedness values between strain TW-4T and closest members were below the threshold value for species delineation. Based on polyphasic taxonomic analyses, strain TW-4T represents novel species in the genus Novosphingobium, for which the name Novosphingobium olei sp. nov. is proposed. The type strain is TW-4T (=KACC 21628T=NBRC 114364T) and strain TNP-2 (=KACC 21629=NBRC 114365) represents an additional strain. Based on new data obtained in this study, it is also proposed to reclassify Novosphingobium stygium as a later heterotypic synonym of Novosphingobium aromaticivorans.

Solirhodobacter olei gen. nov., sp. nov., a nonphotosynthetic bacterium isolated from oil-contaminated soil.

A nonphotosynthetic, Gram-stain-negative, rod-shaped and motile strain, designated Pet-1T, was isolated from oil-contaminated soil collected from Daqing oil field in China. Optimal growth occurred at 37 °C, pH 5.5 and in 1 % (w/v) NaCl. Q-10 was the sole respiratory quinone. The most abundant fatty acid was C18 : 1É·7c/C18 : 1É·6c (67.4 %). The major polar lipids were phosphatidylglycerol, aminolipid, phosphatidylethanolaine, phosphatidycholine, two unidentified lipids and two unidentified phospholipids. The genomic DNA G+C content was 69.3 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that Pet-1T shared the highest similarity (95.1 %) to Rhodobacter vinaykumarii DSM 18714T, followed by Sinorhodobacter populi sk2b1T (95.0 %) and Haematobacter massiliensis CCUG 47968T (95.0 %). In the phylogenetic tree, strain Pet-1T formed a separate branch from the closely related genera Rhodobacter, Pararhodobacter, Defluviimonas and Rhodovulum within the family Rhodobacteraceae. Based on the data from the current polyphasic study, it is proposed that the isolate is a novel species of a novel genus within the family Rhodobacteraceae, with the name Solirhodobacter olei gen. nov., sp. nov. The type strain of the type species is Pet-1T (=KCTC 72074T =CCTCC AB 2018368T).

Sphingobacterium olei sp. nov., isolated from oil-contaminated soil.

A Gram-stain-negative, rod-shaped, non-motile and non-spore-forming bacterium, designated HAL-9T, was isolated from oil-contaminated soil in Daqing oilfield, Heilongjiang Province, PR China. Strain HAL-9T was able to degrade quizalofop-p-ethyl and diclofop-methyl. Growth was observed at 10-35 °C (optimum, 30 °C), pH 6.0-10.0 (optimum, pH 7.0) and salinity of 0 %-5.0 % (w/v; optimum 1.0 %). The results of phylogenetic analysis based on the 16S rRNA gene indicated that strain HAL-9T belongs to the genus Sphingobacterium and showed the highest sequence similarity (98.3 %) to Sphingobacterium alkalisoli Y3L14T, followed by Sphingobacterium mizutaii DSM 11724T (95.1 %) and Sphingobacterium lactis DSM 22361T (95.1 %). Menaquinone-7 (MK-7) was the only isoprenoid quinone. The predominant cellular fatty acids were summed feature 3 (C16 : 1 ω7c and/or C16 : 1 ω6c), iso-C15: 0 and iso-C17 : 0 3-OH. The major polar lipids were phosphatidylethanolamine, three phosphoglycolipids and three unidentified lipids. The draft genome of strain HAL-9T was 5.41 Mb. The G+C content of strain HAL-9T was 40.6 mol%. Furthermore, the average nucleotide identity and in silico DNA-DNA hybridization values between strain HAL-9T and S. alkalisoli Y3L14T were 86.2 % and 32.8 %, respectively, which were below the standard thresholds for species differentiation. On the basis of phenotypic, genotypic and phylogenetic evidence, strain HAL-9T represents a novel species in the genus Sphingobacterium, for which the name Sphingobacterium olei sp. nov. is proposed. The type strain is HAL-9T (=ACCC 61581T=CCTCC AB 2019176T=KCTC 72287T).

Enhanced treatment of the oil-contaminated soil using biosurfactant-assisted washing operation combined with H2O2-stimulated biotreatment of the effluent.

A real crude oil-contaminated soil was treated using a two-step method: biosurfactant-assisted soil washing and the biostimulated biotreating of the effluent. The mixture of surfactin and rhamnolipid could enhance the TPH removal from an oil-contaminated soil (32 g/kg) in the soil washing operation. 86% of TPH was removed from the oil-contaminated soil in the soil washing operation under the mixed biosurfactant (surfactin + rhamnolipid) of 0.6 g/L, the soil/water ratio of 20 w/v%, the temperature of 30 °C, and the washing time of 24 h, leaving an effluent containing 5028 mg/L TPH. The effluent was efficiently biotreated in the bioprocess with 5 g/L acclimate biomass daily stimulated with 0.1 mM H2O2, and the concentrtion of TPH decreased to 26 mg/L within 17 d corresponding a TPH biodegradation over 99%. The biostimulation with H2O2 caused the production of a high amount of peroxidase that could accelerate the biodegradation of TPH. Accordingly, the findings suggest that the biosurfactant-assisted washing operation combined with the H2O2-stimulated biodegradation process could be an enhanced green method for efficient treatment of the heavy oil-contaminated soils.

Growth of zinnia, Italian ryegrass, and alfalfa and their remediation effects in diesel oil-contaminated soils.

Our objective in this study was to compare the growth of zinnia, Italian ryegrass, and alfalfa, and their remediation effects in oil-contaminated soils. The soils were prepared by mixing 2, 4, or 8% diesel oil by weight with soil. The plant height and dry weights of shoots and roots were highest for zinnia in the 2 and 4% oil treatments, and highest for Italian ryegrass in the 8% oil treatment. The reduction ratios in soil total petroleum hydrocarbons concentration (TPH) for 3 plants were lower in the 4 and 8% oil treatments than those in the 2% treatment. The reduction ratios for Italian ryegrass and zinnia contaminated with 2, 4, and 8% diesel oil treatments were significantly higher than those for alfalfa and the non-cultivation treatment at 45 days after sowing, and there were no significant differences in reduction ratios between Italian ryegrass and zinnia. The reduction ratio of soil TPH concentration brought about by zinnia was also comparable to that of Italian ryegrass. Therefore, we conclude that zinnia shows growth and remediation effects that are equivalent to those of Italian ryegrass, in soils contaminated with less than 8% oil.

Relationships between root growth of Zinnia hybrid "profusion orange" flowers and phytoremediation of oil-contaminated soil.

Relationships exist between plant root growth and the phytoremediation of oil-contaminated soils. In a previous study, we demonstrated that zinnia flowers are well suited for the remediation of oil-contaminated soil. In this study, our goal was to quantify the relationship between zinnia root growth and purification of oil-contaminated soils. Three treatments were used: (1) cultivation of zinnia in oil-contaminated soil (contaminated pots), (2) cultivation in non-contaminated soil (non-contaminated pots), and (3) contaminated soil with no cultivation and only irrigation (irrigated pots). Growth of the Zinnia plants, including their roots, was significantly reduced in the contaminated pots compared with the noncontaminated pots. The soil dehydrogenase activity increased between 45 and 90 days after planting in all parts of the contaminated pots, especially the upper parts. The soil total petroleum hydrocarbon (TPH) concentrations in the contaminated pots decreased throughout the study period. Interestingly, the soil dehydrogenase activity increased, and the soil TPH concentration decreased even in lower parts of the pots where there was very little root growth. Therefore, the cultivation of plants can have a remediative effect on oil-contaminated soil even below the depth reached by the plant roots.

Effects of bacterial-feeding nematodes on soil microbial activity and the microbial community in oil-contaminated soil.

Rapid economic development has caused an increase in serious pollution problems due to the ever-increasing use of oil and its products, thus making oil pollution control an urgent task. Studies have shown that large amounts of bacterial-feeding nematodes are present in oil-contaminated soil; their function is as yet unclear. In this experiment, different densities of Caenorhabditis elegans (C. elegans) were inoculated into artificially simulated oil-contaminated soil to examine their effects on microbial activity and the microbial community in oil-contaminated soil. Six treatments were investigated: sterilized oil-contaminated soil as control 1 (FSP), nematode-free soil as control 2 (S), oil-contaminated soil (SP), oil-contaminated soil + 5, 10 or 20 individual C. elegans per gram of dry soil (i.e., SPN5, SPN10, SPN20). Results showed that oil pollution significantly increased the soil basal respiration. However, C. elegans weakened the soil basal respiration to different degrees and soil microbial respiration entropy essentially changed in line with the soil basal respiration. Oil pollution and C. elegans boosted catalase activity in contaminated soil by approximately 64.2-145.1%. Soil urease activity of SPN5, SPN10 and SPN20 was 88.5%, 126.7% and 109.0% stronger, respectively, than that of SP. The inoculation of C. elegans changed the microbial phospholipid fatty acid content in the oil-contaminated soil, including soil bacteria, fungi, actinomycetes, Gram-positive bacteria (G+) and Gram-negative bacteria (G-). Therefore, this research demonstrates that C. elegans can stimulate microbial reproduction in oil-contaminated soil, enhance related soil enzyme activities and regulate soil microbial community structure and diversity, thereby improving the contaminated soil environment and promoting oil degradation.

Sphingomonas oleivorans sp. nov., isolated from oil-contaminated soil.

Strain FW-11T was isolated from oil-contaminated soil from Panjin in Liaoning, China. It was a Gram-stain-negative, aerobic and rod-shaped bacterium. The strain was confirmed to be a member of the genus Sphingomonas based on phylogenetic inference and phenotypic characteristics. The best growth of strain FW-11T occurred at 30 °C and pH 6.0-7.0. The strain was non-spore-forming, catalase-negative and oxidase-negative. The main polar lipids were sphingoglycolipid, phosphatidylcholine, phosphatidylglycerol, diphosphatidylglycerol, phosphatidylethanolamine and unidentified lipids. The cell-wall peptidoglycan of strain FW-11T included alanine, glycine, glutamic acid, aspartic acid and meso-diaminopimelate. The predominant isoprenoid quinones were ubiquinone Q-10 (93.2 %) and Q-9 (6.8 %). The fatty acid profile (>5 %) included C18 : 1ω6c (43.1 %), C16 : 0 (14.6 %), C17 : 1ω6c (14.0 %) and C14 : 0 2-OH (11.1 %). The most similar neighbours of FW-11T were Sphingomonas fennica K101T (97.4 %) and Sphingomonas haloaromaticamans A175T (97.0 %). The average nucleotide identity relatedness values between strain FW-11T and the two type strains (S. fennica K101T and S. haloaromaticamans A175T) were 73.2 and 75.3 %, respectively. The genome size of FW-11T was 3.8 Mbp, comprising 3735 predicted genes with a DNA G+C content of 64.0 mol%. Based on phenotypic, chemotaxonomic and phylogenetic data, strain FW-11T represents a novel species of the genus Sphingomonas, for which the name Sphingomonas oleivorans sp. nov. is proposed. The type strain is FW-11T (=LMG 29274T=HAMBI 3659T).