Biodegradation of petroleum hydrocarbons in soil inoculated with yeasts

Yeast species belonging to the Candida genus were added to the greyish-brown soil of the Apsheron Peninsula under laboratory conditions. The rate of CO2 production was used to estimate the degradation of crude oil, paraffin, cycloparaffin and aromatic hydrocarbons as well as their oxidized products. The rate of hydrocarbon degradation in the soil inoculated with yeast cells was shown to drop down gradually. The effective action on the process of hydrocarbon degradation depended on the special properties of an inoculated population and on the structure of a hydrocarbon. Some yeast species stimulated the degradation of various aromatic hydrocarbons and their oxidized products. Aromatic hydrocarbons were decomposed at a lower rate comparing to their oxidized products. When the soil was inoculated with C. guilliermondii populations, n-hexadecane added to the soil at a concentration of 1% was decomposed within 250-300 days. Field experiments confirmed that crude oil biodegradation was more intensive in the soil inoculated with yeast cells.     

Bacteria--degraders of polycyclic aromatic hydrocarbons, isolated from soil and bottom sediments in salt-mining areas

Fifteen bacterial strains capable of utilizing naphthalene, phenanthrene, and biphenyl as the sole sources of carbon and energy were isolated from soils and bottom sediments contaminated with waste products generated by chemical and salt producing plants. Based on cultural, morphological, and chemotaxonomic characteristics, ten of these strains were identified as belonging to the genera Rhodococcus, Arthrobacter, Bacillus, and Pseudomonas. All ten strains were found to be halotolerant bacteria capable of growing in nutrient-rich media at NaCl concentrations of 1-1.5 M. With naphthalene as the sole source of carbon and energy, the strains could grow in a mineral medium with 1 M NaCl. Apart from being able to grow on naphthalene, six of the ten strains were able to grow on phenanthrene; three strains, on biphenyl; three strains, on octane; and one strain, on phenol. All of the strains were plasmid-bearing. The plasmids of the Pseudomonas sp. strains SN11, SN101, and G51 are conjugative, contain genes responsible for the degradation of naphthalene and salicylate, and are characterized by the same restriction fragment maps. The transconjugants that gained the plasmid from strain SN11 acquired the ability to grow at elevated NaCl concentrations. Microbial associations isolated from the same samples were able to grow at a NaCl concentration of 2.5 M.     

石油烃和酚类物质在土中的生物降解与土壤酶活性

本文通过模拟实验,研究了不同条件下石油烃和酚类物质在土中的降解进程及其与土壤酶活性的关系,并在此基础上,对所述污染物的土地处理提出了若干建议。     

Effect of salinity on the bioremediation of petroleum hydrocarbons in a saline-alkaline soil

Aims: The aim of this paper is to check the effect of salinity on the bioremediation process of petroleum hydrocarbons in the saline‐alkaline soil. Methods and Results: In this study, soil salinity was adjusted to different levels by water leaching method and the bioremediation process was conducted for 28 days. Soil pH increased after leaching and decreased during bioremediation process. At initial time, moderate salinity enhanced the biodegradation and addition of microbial consortium was not effective in enhancing degradation rate of petroleum hydrocarbons. At day of 28 days, higher degradation rate was found in treatments with more leaching times with a maximum value of 42·36%. Dehydrogenase activity increased with the progress of bioremediation and positive correlation was found between dehydrogenase activity and degradation rate of petroleum hydrocarbons. Denaturing gradient gel electrophoresis analysis result showed decreased microbial community diversity with increased salt content. Conclusions: The result suggested that salinity had great impact on bioremediation, and leaching and addition of inoculated consortium were effective in enhancing biodegradation of petroleum hydrocarbons in the saline‐alkaline soil. Significance and Impact of the Study: The result of this study is important for understanding the bioremediation process of petroleum in contaminated soil. New remediation method of petroleum contaminated soil can be developed based on this study.     

石油烃生物降解过程的研究进展

石油烃污染物属于难降解混合物,生物修复已经成为石油烃污染环境的主要修复方法.文中简述了微生物对石油烃的间期适应过程和转运过程,并通过对部分典型石油烃成分的微生物降解机理和代谢路径的梳理和综述,阐释了石油烃生物降解过程中的菌株、基因、代谢路径等研究进展.此外,利用基因工程和代谢工程等手段,可对野生型石油烃降解菌进行改造,...     

Potential of preliminary test methods to predict biodegradation performance of petroleum hydrocarbons in soil

Preliminary tests at different scales such as degradation experiments (laboratory) in shaking flasks, soil columns and lysimeters as well as in situ respiration tests (field) were performed with soil from two hydrocarbon contaminated sites. Tests have been evaluated in terms of their potential to provide information on feasibility, degradation rates and residual concentration of bioremediation in the vadose zone. Sample size, costs and duration increased with experimental scale in the order shaking flasks – soil columns – lysimeter – in situ respiration tests, only time demand of respiration tests was relatively low. First-order rate constants observed in degradation experiments exhibited significant differences between both, different experimental sizes and different soils. Rates were in line with type and history of contamination at the sites, but somewhat overestimated field rates particularly in small scale experiments. All laboratory experiments allowed an estimation of residual concentrations after remediation. In situ respiration tests were found to be an appropriate pre-testing and monitoring tool for bioventing although residual concentrations cannot be predicted from in situ respiration tests. Moreover, this method does not account for potential limitations that might hamper biodegradation in the longer term but only reflects the actual degradation potential when the test is performed.     

Effects of co-occurring aromatic hydrocarbons on degradation of individual polycyclic aromatic hydrocarbons in marine sediment slurries

Rates of polycyclic aromatic hydrocarbon (PAH) degradation and mineralization were influenced by preexposure to alternate PAHs and a monoaromatic hydrocarbon at relatively high (100 ppm) concentrations in organic-rich aerobic marine sediments. Prior exposure to three PAHs and benzene resulted in enhanced [14C]naphthalene mineralization, while [14C]anthracene mineralization was stimulated only by benzene and anthracene preexposure. Preexposure of sediment slurries to phenanthrene stimulated the initial degradation of anthracene. Prior exposure to naphthalene stimulated the initial degradation of phenanthrene but had no effect on either the initial degradation or mineralization of anthracene. For those compounds which stimulated [14C]anthracene or [14C]naphthalene mineralization, longer preexposures (2 weeks) to alternative aromatic hydrocarbons resulted in an even greater stimulation response. Enrichment with individual PAHs followed by subsequent incubation with one or two PAHs showed no alteration in degradation patterns due to the simultaneous presence of PAHs. The evidence suggests that exposure of marine sediments to a particular PAH or benzene results in the enhanced ability of these sediments to subsequently degrade that PAH as well as certain other PAHs. The enhanced degradation of a particular PAH after sediments have been exposed to it may result from the selection and proliferation of specific microbial populations capable of degrading it. The enhanced degradation of other PAHs after exposure to a single PAH suggests that the populations selected have either broad specificity for PAHs, common pathways of PAH degradation, or both.     

Breakdown of low-level total petroleum hydrocarbons (TPH) in contaminated soil using grasses and willows

A phytoremediation study targeting low-level total petroleum hydrocarbons (TPH) was conducted using cool- and warm-season grasses and willows (Salix species) grown in pots filled with contaminated sandy soil from the New Haven Rail Yard, CT. Efficiencies of the TPH degradation were assessed in a 90-day experiment using 20–8.7–16.6 N-P-K water-soluble fertilizer and fertilizer with molasses amendments to enhance phytoremediation. Plant biomass, TPH concentrations, and indigenous microbes quantified with colony-forming units (CFU), were assessed at the end of the study. Switchgrass grown with soil amendments produced the highest aboveground biomass. Bacterial CFU's were in orders of magnitude significantly higher in willows with soil amendments compared to vegetated treatments with no amendments. The greatest reduction in TPH occurred in all vegetated treatments with fertilizer (66–75%) and fertilizer/molasses (65–74%), followed sequentially by vegetated treatments without amendments, unvegetated treatments with amendments, and unvegetated treatments with no amendment. Phytoremediation of low-level TPH contamination was most efficient where fertilization was in combination with plant species. The same level of remediation was achievable through the addition of grasses and/or willow combinations without amendment, or by fertilization of sandy soil.     

Morphological,biochemical and molecular identification of petroleum hydrocarbons biodegradation bacteria isolated from oil polluted soil in Dhahran,Saud Arabia

Accumulation of petroleum hydrocarbon residual considered a major environmental problem in the kingdom of Saudi Arabia cause of intensive efforts for oil detecting. Until now, In situ biodegradation considered the most effective method for petroleum hydrocarbon residual biodegradation. The aim of this study is isolation and identification biodegradable capability bacteria from contaminated sites in Khurais oil field, Dhahran, Saud Arabia via Different morphological and biochemical and molecular methods. Furthermore, degradation level in contaminated liquid medium and soil were evaluated. Three bacterial strains were selected from petroleum-contaminated soils of Khurais oil field depending on their capacity to grow in the existence of hydrocarbon components and identified according to morphological, biochemical. Interestingly, 16S rDNA sequencing fingerprinting results confirmed our bacterial identification as Bacillus subtilis, Pseudomonas aeruginosa and Bacillus cereu. Phyllogenetic tree was constructed and genetic similarity was calculated according to alignments results. Biodegradation patterns for different three isolates were reflected varied degradation ability for three isolates regarding incubation time. Different features were studied for three biodegrading bacterial strains and identified as Bacillus subtilis, Pseudomonas aeruginosa and Bacillus cereus. Remarkable degradation rate % patterns for hydrocarbons residual were recorded for all three isolates with varied.     

Effects of co-occurring aromatic hydrocarbons on degradation of individual polycyclic aromatic hydrocarbons in marine sediment slurries.

Rates of polycyclic aromatic hydrocarbon (PAH) degradation and mineralization were influenced by preexposure to alternate PAHs and a monoaromatic hydrocarbon at relatively high (100 ppm) concentrations in organic-rich aerobic marine sediments. Prior exposure to three PAHs and benzene resulted in enhanced [14C]naphthalene mineralization, while [14C]anthracene mineralization was stimulated only by benzene and anthracene preexposure. Preexposure of sediment slurries to phenanthrene stimulated the initial degradation of anthracene. Prior exposure to naphthalene stimulated the initial degradation of phenanthrene but had no effect on either the initial degradation or mineralization of anthracene. For those compounds which stimulated [14C]anthracene or [14C]naphthalene mineralization, longer preexposures (2 weeks) to alternative aromatic hydrocarbons resulted in an even greater stimulation response. Enrichment with individual PAHs followed by subsequent incubation with one or two PAHs showed no alteration in degradation patterns due to the simultaneous presence of PAHs. The evidence suggests that exposure of marine sediments to a particular PAH or benzene results in the enhanced ability of these sediments to subsequently degrade that PAH as well as certain other PAHs. The enhanced degradation of a particular PAH after sediments have been exposed to it may result from the selection and proliferation of specific microbial populations capable of degrading it. The enhanced degradation of other PAHs after exposure to a single PAH suggests that the populations selected have either broad specificity for PAHs, common pathways of PAH degradation, or both.     

Effects on microbial activity by extraction of indigenous cells from soil slurries

Abstract: Possible effects on the physiological activity and culturability of soil microorganisms by different soil dispersion procedures, and effects on activity caused by extracting bacteria from soil, were investigated. There was no apparent difference in cfu's with dispersion of a silty loam soil and a loamy sand soil with pyrophosphate as compared to dispersion in NaCl. Substrate-induced respiration was reduced in the silty loam soil, and methanol oxidation was reduced in the loamy sand soil with dispersion in pyrophosphate, and the soil pH was irreversibly increased by the treatment. Extracted bacterial fractions had lower numbers of culturable cells as percentage of the total number of bacteria in each fraction, lower respiration rates and no methanol oxidation activity as compared to the soil slurry both before and after extraction. The physiological activity was apparently not affected by the number of cells extracted. This indicates that the increased extraction rate of indigenous soil bacteria obtained by effective disruption of aggregates and detachment of cells from surfaces, only results in increased extraction of cells that have been physiologically changed as a result of the extraction process.     

Biodegradation of petroleum hydrocarbons by filamentous fungi (Aspergillus ustus and Purpureocillium lilacinum) isolated from used engine oil contaminated soil

The use of microorganisms for remediation and restoration of hydrocarbons contaminated soils is an effective and economic solution. The current study aims to find out efficient telluric filamentous fungi to degrade petroleum hydrocarbons pollutants. Six fungal strains were isolated from used engine (UE) oil contaminated soil. Fungi were screened for their ability to degrade crude oil, diesel and UE oil using 2.6-dichlorophenol indophenol (DCPIP). Two isolates were selected, identified and registered at NCBI as Aspergillus ustus HM3.aaa and Purpureocillium lilacinum HM4.aaa. Fungi were tested for their tolerance to different concentration of petroleum oils using radial growth diameter assay. Hydrocarbons removal percentage was evaluated gravimetrically. The degradation kinetic of crude oil was studied at a time interval of 10 days. A.ustus was the most tolerant fungi to high concentration of petroleum oils in solid medium. Quantitative analysis showed that crude oil was the most degraded oil by both isolate; P. lilacinium and A. ustus removed 44.55% and 30.43% of crude oil, respectively. The two fungi were able to degrade, respectively, 27.66 and 21.27% of diesel and 14.39 and 16.00% of UE oil. As compared to the controls, these fungi accumulated high biomass in liquid medium with all petroleum oils. Likewise, crude oil removal rate constant (K) and half-lives (t_1/2) were 0.02 day⁻¹, 34.66 day and 0.015 day⁻¹, 46.21 day for P. lilacinium and A. ustus, respectively. The selected fungi appear interesting for petroleum oils biodegradation and their application for soil bioremediation require scale-up studies.     

Biodegradation of petroleum hydrocarbons under tropical estuarine conditions

The physico-chemical parameters of water samples collected from three points in the Lagos lagoon were studied for 12 months. Safinity varied seasonally but the temperature pH, dissolved O₂, conductivity, NO₃ ^– and HPO₄ ^2– concentrations were relatively constant. There was a direct proportionality between the population density of hydro-carbon-utilizing bacteria and the oil content of water samples. Twelve hydrocarbon-utilizing bacteria were isolated by selective enrichment and characterized as species ofPseudomonas, Alcaligenes, Acinetobacter andBacilius. The organisms grew mainly on long-chain aliphatic hydrocarbons. Laboratory and field biodegradation studies showed both quantitative and qualitative changes in the hydrocarbon content of crude oil due to microbial degradative activities and a faster rate of oil depletion from the Lagos lagoon during the rainy season. The results obtained could offer a predictive model for estimating the rate of disappearance of petroleum hydrocarbons from the tropical estuarine environment.

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Résumé On a étudié les paramètres physicochimiques d'échantillons d'eau récoltés en trols endroits du lagon de Lagos pendant 12 mols. La salinite a varié avec les saisons mais tant la température que le pH, l'oxygène dissous, la conductivité et les teneurs en nitrales et en phosphates sont restés relativement constants. II y avait une proportionnalité directe entre la densité de population des bactéries utilisant les hydrocarbures et le contenu en pétrole des échantilions d'eau. Douze souches de bactéries utilisant les hydrocarbures ont été isolées par enrichissement sélectif et caractérisées au niveau de l'espèce comme desPseudomonas, Alcaligenes, Acinetobacter etBacillus. Ces organismes croissent principalement sur des hydrocarbures aliphatiques à longue chaîne. Des études de biodégradation au laboratoire et sur le terrain ont montré des changements tant quantitatifs que qualitatifs dans le contenu en hydrocarbures du pétroie brut, dus aux activités de dégradation microbienne ainsi qu'un appauvrissement plus rapide en pétrole du lagon de Lagos pendant la saison des pluies. Les résultats obtenus pourraient offrir un modèle prédictif pour l'estimation de la vitesse de disparition des hydrocarbures du pétrole dans l'environnement d'estuaires tropicaux.

     

铁还原菌降解石油烃的研究进展

铁还原菌是指能够利用细胞外Fe(III)作为末端电子受体,通过氧化有机物将Fe(III)还原为Fe(II)微生物的总称。铁还原作用广泛存在于土壤、河流、海洋、地表含水层以及高温高压的地下深部油藏。在厌氧或兼性厌氧条件下,Fe(III)还原耦合有机物的降解,对铁、碳元素的生物地球化学循环具有重要意义。本文介绍了铁还原菌的多样性和铁还原作用机理,综述了铁还原菌在石油烃降解方面的研究进展。此外,还总结了铁还原菌在生物修复中的潜在作用,并对未来的研究方向进行了展望。     

Bacteria which attack petroleum hydrocarbons in a saline medium

Bacterial strains were isolated from California coastal areas which showed the ability to oxidize normal paraffins, iso-paraffins, and aromatic hydrocarbons in a synthetic seawater medium. The ability to utilize a particular hydrocarbon was established not only on the basis of visible bacterial growth but also through a chromatographic technique which was standardized and which could define the amount of each hydrocarbon consumed by the bacteria in a mixture. Some of the strains exhibited vigorous hydrocarbon oxidation when exposed to synthetic mixtures of hydrocarbons as well as crude oil. Under conditions of aeration and agitation, mixed cultures could destroy approximately 50% of a South Louisiana crude oil in a period of 48 hr.     

Removal of polycyclic aromatic hydrocarbons from oil-contaminated Kuwaiti soil

In the uncontaminated farm soil, more than 80% of the supplemented acenaphthene, fluoranthene, and pyrene (100 mg/100 g soil) decreased in 90 days, while ratio of removal was about 20%, 30%, and 0%, respectively, in the Kuwaiti oil-contaminated soil. Simultaneous addition of naphthalene, phenanthrene, and anthrathene (100 mg of each compound/100 g soil) led the acenaphthene to a decrease of about 20% to 45% but not of fluoranthene and pyrene. Addition of the farm soil to the Kuwaiti soil did not enhance the decrease of these three PAHs.     

Methodology for using contaminated soil leachability testing to determine soil cleanup levels at contaminated petroleum underground storage tank (UST) sites

The costs of environmental remediation at leaking petroleum underground storage tank (UST) sites are influenced significantly by soil cleanup levels. The use of conservative generic soil cleanup levels may be inappropriate at some sites contaminated by leaking petroleum USTs. At many contaminated sites, a primary objective of site remediation is long‐term protection of water resources (e.g., groundwater) from pollution. Leaching of pollutants from residual soil contamination to groundwater is a primary consideration in establishing site‐specific soil cleanup levels at fuel‐contaminated sites. The use of laboratory soil leachability testing methods may be useful in objectively evaluating the leaching potential of contaminants from residual soil contamination and estimating potential groundwater impacts. Developing soil cleanup levels that are protective of water resources must include a technically sound integration of site‐specific soil leachability data and contaminant attenuation factors. Evaluation of the leaching potentials of soil contaminants may also provide essential supplementary information for other site characterization methods that may be used to evaluate risks to human health. Contaminant leachability testing of soils may provide a cost‐effective and technically based method for determining soil cleanup levels that are protective of groundwater resources at contaminated petroleum UST sites.     

Potential nitrification in alum-treated soil slurries amended with poultry manure

Alum is used to reduce environmental pollutants in poultry production. Alum decreases NH3 volatilization and increases total N and NH4+-N compared to untreated poultry manure. Nitrification in poultry wastes could therefore be stimulated due to higher NH4+ concentrations or could be inhibited because the soil environment is acidified. A 10-day laboratory study was conducted to study potential nitrification rates in soil slurries (20 g soil in 150 ml water) amended with 2.0 g alum-treated poultry manure. Fecal bacteria, NH4+, NO2-, NO3-, orthophosphate, pH, and NH3 were measured at 2-day intervals. Alum significantly reduced fecal bacteria concentrations through day 6. Water-soluble P was reduced 82% by day 10. Alum-treated manure had significantly increased NH4+ concentrations by day 8 and 10, and significantly decreased NO2- and NO3- concentrations by days 6-10. Alum's effect on potential nitrification was inhibitory in the soil environment. Slurries with alum-treated poultry manure had reduced nitrification rates, fecal bacteria, and soluble P. Therefore, in addition to reducing P loss, alum could temporarily reduce the risk for environmental pollution from land-applied manures in terms of both NO3- and fecal bacteria loss.