Effect of Modified Fenton’s Reaction on Microbial Activity and Removal of PAHs in Creosote Oil Contaminated Soil

This study describes the removal of polycyclic aromatic hydrocarbons (PAHs) from creosote oil contaminated soil by modified Fenton’s reaction in laboratory-scale column experiments and subsequent aerobic biodegradation of PAHs by indigenous bacteria during incubation of the soil. The effect of hydrogen peroxide addition for 4 and 10 days and saturation of soil with H₂O₂ on was studied. In both experiments the H₂O₂ dosage was 0.4 g H₂O₂/g soil. In completely H₂O₂−saturated soil the removal of PAHs (44% within 4 days) by modified Fenton reaction was uniform over the entire soil column. In non-uniformly saturated soil, PAH removal was higher in completely saturated soil (52% in 10 days) compared to partially saturated soil, with only 25% in 10 days. The effect of the modified Fenton’s reaction on the microbial activity in the soil was assessed based on toxicity tests towards Vibrio fischeri, enumeration of viable and dead cells, microbial extracellular enzyme activity, and oxygen consumption and carbon dioxide production during soil incubation. During the laboratory-scale column experiments, the toxicity of column leachate towards Vibrio fischeri increased as a result of the modified Fenton’s reaction. The activities of the microbial extracellular enzymes acetate- and acidic phosphomono-esterase were lower in the incubated modified Fenton’s treated soil compared to extracellular enzyme activities in untreated soil. Abundance of viable cells was lower in incubated modified Fenton treated soil than in untreated soil. Incubation of soil in serum bottles at 20 °C resulted in consumption of oxygen and formation of carbon dioxide, indicating aerobic biodegradation of organic compounds. In untreated soil 20–30% of the PAHs were biodegraded during 2 months of incubation. Incubation of chemically treated soil slightly increased PAH-removal compared to PAH-removal in untreated soil.     

Free hydroxyl radical is not involved in an important reaction of lignin degradation by Phanerochaete chrysosporium Burds.

Hydroxyl radical (HO.) has been implicated in the degradation of lignin by Phanerochaete chrysosporium. This study assessed the possible involvement of HO. in degradation of lignin substructural models by intact cultures and by an extracellular ligninase isolated from the cultures. Two non-phenolic lignin model compounds [aryl-C(alpha)HOH-C(beta)HR-C(gamma)H2OH, in which R = aryl (beta-1) or R = O-aryl (beta-O-4)] were degraded by cultures, by the purified ligninase, and by Fenton's reagent (H2O2 + Fe2+), which generates HO.. The ligninase and the cultures formed similar products, derived via an initial cleavage between C(alpha) and C(beta) (known to be an important biodegradative reaction), indicating that the ligninase is responsible for model degradation in cultures. Products from the Fenton degradation were mainly polar phenolics that exhibited little similarity to those from the biological systems. Mass-spectral analysis, however, revealed traces of the same products in the Fenton reaction as seen in the biological reactions; even so, an 18O2-incorporation study showed that the mechanism of formation differed. E.s.r. spectroscopy with a spin-trapping agent readily detected HO. in the Fenton system, but indicated that no HO. is formed during ligninase catalysis. We conclude, therefore that HO. is not involved in fungal C(alpha)-C(beta) cleavage in the beta-1 and beta-O-4 models and, by extension, in the same reaction in lignin.     

Enhanced transformation of polycyclic aromatic hydrocarbons using a combined Fenton's reagent,microbial treatment and surfactants

The potential for using Fenton's reagent (H₂O₂ + Fe²⁺) as an advanced oxidation pretreatment process to enhance microbial transformation of two model polycyclic aromatic hydrocarbons, anthracene and benzo[a]pyrene, in an aqueous system was evaluated. Fenton's reagent at a concentration of 0.5% H₂O₂ and 10 mM Fe²⁺ (molar ratio, 15:1) was most effective in transforming anthracene at pH 4. Application of non-ionic surfactants during Fenton's pre-treatment was found to be more effective in the transformation of both anthracene and benzo[a]pyrene. The extent of removal of substrates by a combined Fenton's–biotreatment was 2–4 times higher than with Fenton's treatment or biotreatment alone. In a chemical–biological treatment train, 48 h of Fenton's pre-treatment in the presence of a non-ionic surfactant, followed by 7 days of biological treatment resulted in 80–85% removal of PAHs (100 ppm). Electronic Publication     

酚顿试剂对竹林土壤中酚类化合物的降解作用

酚类化合物是农业和生态系统中主要的化感物质之一,大量积累在土壤中,抑制作物和林下植物生长,导致农作物减产、连作障碍和自然生态环境破坏。用过氧化氢(H2O2)与硫酸亚铁(Fe2 )所组成的酚顿试剂研究了化学氧化法对化感物质(对香豆酸、对羟基苯甲酸和胡桃醌)、竹林土壤提取物及竹林土壤中对香豆酸的降解作用。过氧化氢与硫酸亚铁的摩尔比为15:1的酚顿试剂,过氧化氢与酚类物质浓度比为8:1时,对酚类物质的降解效率最高。以8×10-3mol/L(0.028%)的H2O2,5.4×10-4mol/L(0.075%)的FeSO4和10-3mol/L的酚类物质组成的反应体系中,反应10min和30min后,对香豆酸的降解率分别为55%和74%;对羟基苯甲酸的降解率在10min时达90%以上;而胡桃醌在10min时已经完全被降解。酚顿试剂处理土壤酚类提取物时,可使其中主要的化感物质对香豆酸降解75%。用含0.1%和1%H2O2的酚顿试剂处理竹(Bambusa chungii)林土壤,土壤对香豆酸的降解率分别为32%和37%。竹林土壤中存在过氧化氢酶和过氧化物酶活性,但没有检测到超氧化物歧化酶活性。土壤中的过氧化氢酶和过氧化物酶可能迅速分解外加的过氧化氢,一方面缩短过氧化氢处理的作用时间和降低降解效率,另一方面可分解过剩的过氧化氢。这说明酚顿试剂是降解土壤和培养液中有害化感物质的有效化学氧化剂。     

Long-term simulation of in situ biostimulation of polycyclic aromatic hydrocarbon-contaminated soil

A continuous-flow column study was conducted to evaluate the long-term effects of in situ biostimulation on the biodegradation of polycyclic aromatic hydrocarbons (PAHs) in soil from a manufactured gas plant site. Simulated groundwater amended with oxygen and inorganic nutrients was introduced into one column, while a second column receiving unamended groundwater served as a control. PAH and dissolved oxygen (DO) concentrations, as well as microbial community profiles, were monitored along the column length immediately before and at selected intervals up to 534?days after biostimulation commenced. Biostimulation resulted in significantly greater PAH removal than in the control condition (73% of total measured PAHs vs. 34%, respectively), with dissolution accounting for a minor amount of the total mass loss (~6%) in both columns. Dissolution was most significant for naphthalene, acenaphthene, and fluorene, accounting for >20% of the total mass removed for each. A known group of PAH-degrading bacteria, 'Pyrene Group 2' (PG2), was identified as a dominant member of the microbial community and responded favorably to biostimulation. Spatial and temporal variations in soil PAH concentration and PG2 abundance were strongly correlated to DO advancement, although there appeared to be transport of PG2 organisms ahead of the oxygen front. At an estimated oxygen demand of 6.2?mg O(2)/g dry soil and a porewater velocity of 0.8?m/day, it took between 374 and 466?days for oxygen breakthrough from the 1-m soil bed in the biostimulated column. This study demonstrated that the presence of oxygen was the limiting factor in PAH removal, as opposed to the abundance and/or activity of PAH-degrading bacteria once oxygen reached a previously anoxic zone.     

Characterization of growing microorganisms in soil by stable isotope probing with H218O

A new approach to characterize growing microorganisms in environmental samples based on labeling microbial DNA with H(2)(18)O is described. To test if sufficient amounts of (18)O could be incorporated into DNA to use water as a labeling substrate for stable isotope probing, Escherichia coli DNA was labeled by cultivating bacteria in Luria broth with H(2)(18)O and labeled DNA was separated from [(16)O]DNA on a cesium chloride gradient. Soil samples were incubated with H(2)(18)O for 6, 14, or 21 days, and isopycnic centrifugation of the soil DNA showed the formation of two bands after 6 days and three bands after 14 or 21 days, indicating that (18)O can be used in the stable isotope probing of soil samples. DNA extracted from soil incubated for 21 days with H(2)(18)O was fractionated after isopycnic centrifugation and DNA from 17 subsamples was used in terminal restriction fragment length polymorphism (TRFLP) analysis of bacterial 16S rRNA genes. The TRFLP patterns clustered into three groups that corresponded to the three DNA bands. The fraction of total fluorescence contributed by individual terminal restriction fragments (TRF) to a TRFLP pattern varied across the 17 subsamples so that a TRF was more prominent in only one of the three bands. Labeling soil DNA with H(2)(18)O allows the identification of newly grown cells. In addition, cells that survive but do not divide during an incubation period can also be characterized with this new technique because their DNA remains without the label.     

Iron and dioxygen chemistry is an important route to initiation of biological free radical oxidations: an electron paramagnetic resonance spin trapping study

Iron can be a detrimental catalyst in biological free radical oxidations. Because of the high physiological ratio of [O2]/[H2O2] (> or = 10(3)), we hypothesize that the Fenton reaction with pre-existing H2O2 is only a minor initiator of free radical oxidations and that the major initiators of biological free radical oxidations are the oxidizing species formed by the reaction of Fe2+ with dioxygen. We have employed electron paramagnetic resonance spin trapping to examine this hypothesis. Free radical oxidation of: 1) chemical (ethanol, dimethyl sulfoxide); 2) biochemical (glucose, glyceraldehyde); and 3) cellular (L1210 murine leukemia cells) targets were examined when subjected to an aerobic Fenton (Fe2+ + H2O2 + O2) or an aerobic (Fe2+ + O2) system. As anticipated, the Fenton reaction initiates radical formation in all the above targets. Without pre-existing H2O2, however, Fe2+ and O2 also induce substantial target radical formation. Under various experimental ratios of [O2]/[H2O2] (1-100 with [O2] approximately 250 microM), we compared the radical yield from the Fenton reaction vs. the radical yield from Fe2+ + O2 reactions. When [O2]/[H2O2] < 10, the Fenton reaction dominates target molecule radical formation; however, production of target-molecule radicals via the Fenton reaction is minor when [O2]/[H2O2] > or = 100. Interestingly, when L1210 cells are the oxidation targets, Fe2+ + O2 is observed to be responsible for formation of nearly all of the cell-derived radicals detected, no matter the ratio of [O2]/[H2O2]. Our data demonstrate that when [O2]/[H2O2] > or = 100, Fe2+ + O2 chemistry is an important route to initiation of detrimental biological free radical oxidations.     

An electrochemical biosensor for the detection of tyrosine oxidation induced by Fenton reaction

A simple and sensitive electrochemical biosensor was used to detect tyrosine oxidation induced by hydroxyl radicals generated by Fenton reaction (Fe(2+)/H(2)O(2)). Poly(glu, tyr) (4:1) peptides were immobilized on indium tin oxide (ITO) electrode surface via layer-by-layer assembly technique, and Os(bpy)(3)(2+)-mediated tyrosine oxidation current was employed as the signal reporter of the biosensor. It was found that the electrochemical signal of the peptide decreased markedly after incubation with Fenton reagents. Interestingly, L-dopa, the oxidation product of tyrosine, was likely to form complexes with Fe(III), which could suppress the electro-oxidation of L-dopa and resulted in decrease of current response. Our results indicate that the peptide damage involved two steps and was a second-order reaction. X-ray photoelectron spectroscopy was used to quantitatively determine nitrogen elemental percentage on peptide-coated electrode surface, which eliminated the possibility that signal decrease was caused by peptide backbone cleavage. Moreover, the lowest concentration of Fenton reagents that could be detected was 10 μM Fe(2+) or H(2)O(2), similar to the level in vivo. We suggest that the biosensor can be used to detect protein damage induced by Fenton reaction.     

秸秆预处理对土壤微生物量及呼吸活性的影响

冬小麦秸秆经8．0g·L^-1H2O2(pH11．0)溶液、12．5g·L^-1 NaOH溶液或H2SO4溶液浸泡8h并80℃烘干后,与无机N一起加入土壤,进行室内25℃恒温培养试验,在不同时间测定土壤微生物量C、N和CO2释放速率。结果表明,培养前期,秸秆预处理使土壤微生物量C数量增加了1．0～1．4倍,但降低了土壤微生物的呼吸活性;培养后期,NaOH和H2SO4处理使土壤微生物量C分别下降了28％和42％,但增加了土壤微生物的呼吸活性;H2O2处理则使土壤微生物量N增加90％;土壤微生物区系中的真菌比例在不同时刻有所增加,表明将秸秆预处理后施入土壤,将对土壤中微生物数量和呼吸活性产生一定影响。     

Is *OH the active Fenton intermediate in the oxidation of ethanol?

A re-examination of the data of Rush and Koppenol (J. Inorg. Biochem. 29 (1987) 199) on the competitive oxidation of C2H5OH and Fe2+ by Fenton's reagent shows that the ratio of the rate constants of the two reactions is 3.2 and not 6.3. The significance of this finding is that it is not possible to identify the active intermediate in the Fenton reaction with the *OH radical.     

Pathways for extracellular Fenton chemistry in the brown rot basidiomycete Gloeophyllum trabeum

The brown rot fungus Gloeophyllum trabeum uses an extracellular hydroquinone-quinone redox cycle to reduce Fe(3+) and produce H(2)O(2). These reactions generate extracellular Fenton reagent, which enables G. trabeum to degrade a wide variety of organic compounds. We found that G. trabeum secreted two quinones, 2,5-dimethoxy-1,4-benzoquinone (2,5-DMBQ) and 4,5-dimethoxy-1,2-benzoquinone (4,5-DMBQ), that underwent iron-dependent redox cycling. Experiments that monitored the iron- and quinone-dependent cleavage of polyethylene glycol by G. trabeum showed that 2,5-DMBQ was more effective than 4,5-DMBQ in supporting extracellular Fenton chemistry. Two factors contributed to this result. First, G. trabeum reduced 2,5-DMBQ to 2,5-dimethoxyhydroquinone (2,5-DMHQ) much more rapidly than it reduced 4,5-DMBQ to 4,5-dimethoxycatechol (4,5-DMC). Second, although both hydroquinones reduced ferric oxalate complexes, the predominant form of Fe(3+) in G. trabeum cultures, the 2,5-DMHQ-dependent reaction reduced O(2) more rapidly than the 4,5-DMC-dependent reaction. Nevertheless, both hydroquinones probably contribute to the extracellular Fenton chemistry of G. trabeum, because 2,5-DMHQ by itself is an efficient reductant of 4,5-DMBQ.     

Bioremediation of soil contaminated with native polycyclic aromatic hydrocarbons from unburnt coal using an in-vessel composting method

The present study was undertaken to investigate the removal of 16 polycyclic aromatic hydrocarbons (PAHs) listed as priority pollutants by the US Environmental Protection Agency (USEPA-PAHs) that are found in coal-contaminated soil during simulated in-vessel composting. Contaminated soil (S) was mixed with green waste (W) in the ratios of 3:1, 1:1, and 1:3 under neutral and acidic soil conditions in laboratory-scale composting reactors. The highest removal efficiency of total 16 USEPA-PAHs (71.88%) was observed in S:W ratio of 1:1 and neutral soil treatment with the removal rate constant of 0.0106 day^?1. Results found that the S:W ratio significantly influenced the removal of PAHs during composting but not the initial soil pH. The results of this research suggest that composting is a feasible and appropriate technology to remediate soil contaminated by coal-native PAH.     

Interaction between the Microbial Community and Invading Escherichia coli O157:H7 in Soils from Vegetable Fields

The survival of Escherichia coli O157:H7 in soils can contaminate vegetables, fruits, drinking water, etc. However, data on the impact of E. coli O157:H7 on soil microbial communities are limited. In this study, we monitored the changes in the indigenous microbial community by using the phospholipid fatty acid (PLFA) method to investigate the interaction of the soil microbial community with E. coli O157:H7 in soils. Simple correlation analysis showed that the survival of E. coli O157:H7 in the test soils was negatively correlated with the ratio of Gram-negative (G⁻) to Gram-positive (G⁺) bacterial PLFAs (G⁻/G⁺ ratio). In particular, levels of 14 PLFAs were negatively correlated with the survival time of E. coli O157:H7. The contents of actinomycetous and fungal PLFAs in the test soils declined significantly (P, <0.05) after 25 days of incubation with E. coli O157:H7. The G⁻/G⁺ ratio declined slightly, while the ratio of bacterial to fungal PLFAs (B/F ratio) and the ratio of normal saturated PLFAs to monounsaturated PLFAs (S/M ratio) increased, after E. coli O157:H7 inoculation. Principal component analysis results further indicated that invasion by E. coli O157:H7 had some effects on the soil microbial community. Our data revealed that the toxicity of E. coli O157:H7 presents not only in its pathogenicity but also in its effect on soil microecology. Hence, close attention should be paid to the survival of E. coli O157:H7 and its potential for contaminating soils.     

Functional diversity changes of microbial communities along a soil aquifer for reclaimed water recharge

The physiochemical and functional diversity of soil-attached microorganisms was investigated using a stabilized laboratory-scale soil aquifer treatment (SAT) system. In this system, reclaimed water after ozonation was used as the feed water, and 60% dissolved organic carbon was removed by the unsaturated vadose layer in 0.8?days. Soil biomass (volatile solids, phospholipid extraction) and functional diversity significantly decreased from the unsaturated vadose layer to the saturated aquifer, where they maintained the same level. Using principal components analysis based on substrate utilization pattern, the vadose layer soil sample was clearly separated from the saturated layer samples. Exceptionally, the oxidation rates of esters remained stable during SAT, indicating the purification potential on certain recalcitrant organic compounds in the saturated aquifer given an adequate retention time. Correlation analysis revealed that organic carbon was the key limiting factor for microbial biomass and activity, especially for tyrosine-like aromatic proteins and soluble microbial byproduct-like materials.     

Exposure of nuclear medicine patients to ionizing radiation is associated with rises in HPRT- mutant frequency in peripheral T-lymphocytes

When Chinese hamster fibroblasts were exposed to hydrogen peroxide or to a system consisting of xanthine oxidase and hypoxanthine, which generates superoxide anion plus hydrogen peroxide, sister-chromatid exchanges (SCEs) were formed in a dose-dependent manner. When the iron-complexing agent o-phenanthroline was present in the medium, however, the production of these SCEs was completely inhibited. This fact indicates that the Fenton reaction: Fe2+ + H2O2----OH0 + OH- + Fe3+ is responsible for the production of SCEs. When O2- and H2O2 were generated inside the cell by incubation with menadione, the production of SCE was prevented by co-incubation with copper diisopropylsalicylate, a superoxide dismutase mimetic agent. The most likely role of O2- is as a reducing agent of Fe3+: O2- + Fe3+----Fe2+ + O2, so that the sum of this and the Fenton reaction, i.e., the iron-catalyzed Haber-Weiss reaction, provides an explanation for the active oxygen species-induced SCE: H2O2 + O2(-)----OH- + OH0 + O2. According to this view, the OH radical thus produced is the agent which ultimately causes SCE. These results are discussed in comparison with other mechanisms previously proposed for induction of SCE by active oxygen species.     

Pulse radiolysis and steady-state analyses of the reaction between hydroethidine and superoxide and other oxidants

Hydroethidine (HE) is a cell-permeable probe used for the intracellular detection of superoxide. Here, we report the direct measurement of the rate constant between hydroethidine and superoxide radical anion using the pulse radiolysis technique. This reaction rate constant was calculated to be ca. 2 x 10(6) M(-1) s(-1) in water:ethanol (1:1) mixture. The spectral characteristics of the intermediates indicated that the one-electron oxidation product of HE was different from the one-electron reduction product of ethidium (E+). The HPLC-electrochemical measurements of incubation mixtures containing HE and the oxygenated Fenton's reagent (Fe2+/DTPA/H2O2) in the presence of aliphatic alcohols or formate as a superoxide generating system revealed 2-OH-E+ as a major product. Formation of 2-OH-E+ by the Fenton's reagent without additives was shown to be superoxide dismutase-sensitive and we attribute the formation of superoxide radical anion to the one-electron reduction of oxygen by the DTPA-derived radical. Addition of tert-butanol, DMSO, and potassium bromide to the Fenton's system caused inhibition of 2-OH-E+ formation. Results indicate that reducing and oxidizing radicals have differential effects on the formation of 2-OH-E+.     

石油添加剂甲基叔丁基醚的污染治理技术研究进展

甲基叔丁基醚是一种石油添加剂,广泛应用于中高档汽油中.其对环境造成的污染和对人体健康造成的危害已日益引起人们的高度重视.本文综述了近年来国外有关甲基叔丁基醚的污染治理技术研究进展,主要是高级氧化技术和微生物降解.已用于处理甲基叔丁基醚的高级氧化技术包括;多相光催化氧化法、紫外光加强的过氧化氢氧化法、臭氧法与臭氧-过氧化氢联合氧化法、超声法与超声-臭氧联合氧化法、芬顿法与光芬顿氧化法、氧气的还原性活化和水的γ射线辐射.微生物降解主要涉及有氧代谢和无氧代谢两大途径.     

An electrochemical biosensor for analysis of Fenton-mediated oxidative damage to BSA using poly-o-phenylenediamine as electroactive probe

A sensitive electrochemical procedure based on bovine serum albumin (BSA)/poly-o-phenylenediamine (PoPD)/carbon-coated nickel (C-Ni) nanobiocomposite film modified glassy carbon electrode (BSA/PoPD/C-Ni/GCE) has been developed to explore the electrochemical detection of BSA damage induced by hydroxyl radical. It is the first time that the electrochemical method has been applied for the analysis of Fenton-mediated oxidative damage to proteins. The hydroxyl radical was generated by Fenton reaction (Fe(2+)/H(2)O(2)), which was also validated by ultraviolet-visible (UV-vis) spectroscopy. The decrease in intensity of the PoPD oxidation signals was used as an indicator for the detection of BSA damage. Damage to BSA was also validated by horizontal Attenuation Total Reflection Fourier Transform Infra-red (ATR-FTIR) spectroscopy and the change of protein carbonyl group content achieved by UV-vis spectroscopy. Effects of H(2)O(2) concentration, the ratio of Fe(2+) and H(2)O(2) and incubation time on BSA damage were examined. Protections of BSA from damage by antioxidants were also investigated. These conclusions demonstrated that the proposed electrochemical method is expected to the further application for protein damage studies.     

ROS-challenged keratinocytes as a new model for oxidative stress-mediated skin diseases

In the current study, the effects of the reactive oxygen species (ROS) generator 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH) on extracellular and intracellular ROS production in human keratinocytes (HACAT) were studied. AAPH is a water-soluble compound able to generate ROS at known and constant rates at 37°C. The short treatment (2 h) with AAPH brought a significant dose-dependent increase in NADPH oxidase activity in intact keratinocytes. The long-term treatment (24 h) with AAPH led to a persistent increase in NADPH oxidase activity for up to 48 hour following the AAPH removal from cell incubation medium. ROS and nitric oxide levels, lipoperoxidation, intracellular calcium, mitochondrial superoxide production, and membrane potential were significantly modified in AAPH-treated HACAT. Superoxide dismutase (SOD) and/or catalase addition to HACAT revealed that untreated keratinocytes produce mostly superoxide anion (O ₂ ⁻), while AAPH-treated keratinocytes overproduce hydrogen peroxide (H ₂O ₂) in extracellular medium. H ₂O ₂ is particularly stable and plays important roles in several cell signaling pathways. Taken together, our findings suggest a cost-effective and easily reproducible in vitro model of stressed human keratinocytes releasing significantly elevated ROS amounts in extracellular medium with respect to control keratinocytes. The possible application of the proposed model for keratinocytes-melanocytes cross-talk studies is also suggested. The model of AAPH-stressed human keratinocytes described here can represent a useful tool for redox cross-talk studies between keratinocytes and other skin cell types, and applied for researches regarding skin pathologies associated with oxidative stress.