Bioremediation of petroleum hydrocarbon-contaminated soil by composting in biopiles

Composting of contaminated soil in biopiles is an ex situ technology, where organic matter such as bark chips are added to contaminated soil as a bulking agent. Composting of lubricating oil-contaminated soil was performed in field scale ( [Formula: see text] m(3)) using bark chips as the bulking agent, and two commercially available mixed microbial inocula as well as the effect of the level of added nutrients (N,P,K) were tested. Composting of diesel oil-contaminated soil was also performed at one level of nutrient addition and with no inoculum. The mineral oil degradation rate was most rapid during the first months, and it followed a typical first order degradation curve. During 5 months, composting of the mineral oil decreased in all piles with lubrication oil from approximately 2400 to 700 mg (kg dry w)(-1), which was about 70% of the mineral oil content. Correspondingly, the mineral oil content in the pile with diesel oil-contaminated soil decreased with 71% from 700 to 200 mg (kg dry w)(-1). In this type of treatment with addition of a large amount of organic matter, the general microbial activity as measured by soil respiration was enhanced and no particular effect of added inocula was observed.     

Short-term effects of diesel fuel on rhizosphere microbial community structure of native plants in Yangtze estuarine wetland

Purpose

In this work, short-term effects of diesel fuel on Huangpu?CYangtze estuarine wetland soil microbial community structure were studied under simulated conditions through phospholipid fatty acids (PLFAs) analysis. Four native plant species, bulrush (Scirpus tripueter), galingale (Cyperus rotundus), wildrice (Zizania latifolia), and reed (Phragmites australis) were tested in the experiments.

Method

In the pot experiment, 20?g rhizosphere soils were mixed with 20?g diesel-blended soils. The concentration of total petroleum hydrocarbon was 16,000?mg/kg. All pots were incubated for 14?days in dark at 28°C and watered with 12?mL sterile distilled water to keep a liquid level. Microbial activity of the samples was assessed by hydrolysis of fluorescein diacetate. Measurements of soil PLFAs and analysis on gas chromatography were performed.

Results

The microbial activity in the samples of reed was highest after the exposure. In all samples, the common PLFA was straight-chain saturated fatty acid (SFA) and monounsaturated fatty acid (MUFA). After the exposure the relative abundance of MUFA and polyunsaturated fatty acid decreased by 20%, and the relative abundance of straight-chain SFA increased by 20%. The results of diversity and PCA indicated that the effect of diesel pollutant on the microbial community was far stronger than the root effect and the reed roots enhanced the tolerance of soil microorganisms to diesel significantly.

Conclusions

All results showed that the soil microbial community structure differed significantly with the exposure to diesel. In reed rhizosphere, the soil microorganisms exhibited a strong resistance to diesel fuel. It confirmed that the root of reed improved the biodegradation ability of soil microorganisms for diesel pollutants and they could be reasonably matched to cure and restore the ecological environment of oil-contaminated wetlands.     

Effect of hydrocarbon pollution on the microbial properties of a sandy and a clay soil

The aim of this work was to ascertain the effects of different types of hydrocarbon pollution on soil microbial properties and the influence of a soil's characteristics on these effects. For this, toxicity bioassays and microbiological and biochemical parameters were studied in two soils (one sandy and one clayey) contaminated at a loading rate of 5% and 10% with three types of hydrocarbon (diesel oil, gasoline and crude petroleum) differing in their volatilisation potential and toxic substance content. Soils were maintained under controlled conditions (50-70% water holding capacity, and room temperature) for six months and several microbiological and toxicity parameters were monitored 1, 60, 120 and 180 days after contamination. The toxic effects of hydrocarbon contamination were greater in the sandy soil. Hydrocarbons inhibited microbial biomass, the greatest negative effect being observed in the gasoline-polluted sandy soil. In both soils crude petroleum and diesel oil contamination increased microbial respiration, while gasoline had little effect on this parameter, especially in the sandy soil. In general, gasoline had the highest inhibitory effect on the hydrolase activities involved in N, P or C cycles in both soils. All contaminants inhibited hydrolase activities in the sandy soil, while in the clayey soil diesel oil stimulated enzyme activity, particularly at the higher concentration. In both soils, a phytotoxic effect on barley and ryegrass seed germination was observed in the contaminated soils, particularly in those contaminated with diesel or petroleum.     

用于石油污染土壤降解的高效降解菌的筛选及其降解条件优化

经过富集、分离优选出高效石油降解菌L-1,根据形态观察和生理生化特征初步鉴定为琼氏不动杆菌;采用单因素花盆实验模拟微生物原位修复并对其降解条件进行优化。结果表明,将高效石油降解菌应用于修复石油污染土壤,适宜接种量、表面活性剂浓度、CNP比、翻耕频率分别为15%、0.1%、100∶10∶1和1 d 1次;在该降解条件下修复28 d,可达到16.80%的石油降解率,远远高于土著微生物6.92%的降解率。     

Fenton氧化技术处理稠油污染土壤

利用Fenton氧化技术对稠油污染土壤进行氧化处理,分析对后续微生物修复的促进作用。向1 000 g石油类含量为8%的稠油污染土壤中加入10.0 mL 18 mmol/L Fe2+溶液与10.0 mL 30%H2O2,反应时间为2 h。氧化处理后土壤中石油烃的总去除率可达到31.38%,胶质去除率为45.22%,沥青质去除率为51.26%,胶质的分子量由1 841下降到1 472,沥青质的分子量由5 831下降到5 073。Fenton氧化可使土壤酶活、各类微生物的数量及呼吸强度有不同程度的下降,但在氧化后30 d内,土壤各类微生物数量都超过了原有水平,其中细菌数量最高达到9.84×105CFU/g,是氧化前的数量的1.57倍。以上实验结果表明,Fenton氧化可以有效去除土壤中胶质和沥青质,并且使土壤中微生物的生长速率加快。因此,Fenton氧化能够促进后续的微生物修复。     

石油污染土壤的生物修复室内模拟实验研究

在实验室模拟的条件下,利用从克拉玛依的石油污染土壤中筛选出的4株高效降解菌,以石油烃降解率、脱氢酶活性、呼吸强度、微生物量碳氮和土壤毒性作为评价指标,研究不加生物菌剂不翻耕、不加生物菌剂翻耕、加生物菌剂不翻耕、加生物菌剂翻耕、加固定化菌剂不翻耕和加固定化菌剂翻耕6种不同实验条件对石油污染土壤修复的效果。结果表明,在63 d的修复过程中,加固定化菌剂翻耕实验F组的石油去除率达到了78.7%,比不加生物菌剂不翻耕实验A组的石油去除率提高了49.5%。随着土壤毒性逐渐降低,玉米(Zea mays L.)和赤子爱胜蚓(Eisenia fetida)可以在F组土壤中良好的生长,达到了修复的效果。     

Insight into the dynamic microbial community and core bacteria in composting from different sources by advanced bioinformatics methods

Microbial communities are important for high composting efficiency and good quality composts. This study was conducted to compare the changes of physicochemical and bacterial characteristics in composting from different raw materials, including chicken manure (CM), duck manure (DM), sheep manure (SM), food waste (FW), and vegetable waste (VW). The role and interactions of core bacteria and their contribution to maturity in diverse composts were analyzed by advanced bioinformatics methods combined sequencing with co-occurrence network and structural equation modeling (SEM). Results indicated that there were obviously different bacterial composition and diversity in composting from diverse sources. FW had a low pH and different physiochemical characteristics compared to other composts but they all achieved similar maturity products. Redundancy analysis suggested total organic carbon, phosphorus, and temperature governed the composition of microbial species but key factors were different in diverse composts. Network analysis showed completely different interactions of core bacterial community from diverse composts but Thermobifida was the ubiquitous core bacteria in composting bacterial network. Sphaerobacter and Lactobacillus as core genus were presented in the starting mesophilic and thermophilic phases of composting from manure (CM, DM, SM) and municipal solid waste (FW, VW), respectively. SEM indicated core bacteria had the positive, direct, and the biggest (>?80%) effects on composting maturity. Therefore, this study presents theoretical basis to identify and enhance the core bacteria for improving full-scale composting efficiency facing more and more organic wastes.

     

典型禾本科植物对石油污染土壤的修复作用

石油类污染物对土壤生态系统的结构与功能造成了较为严重的破坏,影响植物的生长,甚至直接影响到人类健康。选择典型禾本科植物-高粱和玉米,通过盆栽实验,种植于模拟石油污染的土壤中,植物成熟收割后,通过测定土壤中总石油烃的含量,植物体中多环芳烃和直链烷烃的含量,研究高粱和玉米对石油污染土壤的修复作用。结果显示:在种植高粱、玉米后,土壤中总石油烃含量明显降低,并且在收获的高粱、玉米植物体中直链烷烃和多环芳烃含量明显高于空白对照组(未检出)。说明高粱、玉米对石油烃具有一定的去除作用,且高粱对土壤中石油烃的去除作用高于玉米;高粱、玉米对土壤中的多环芳烃和直链烷烃具有一定的积累与富集作用。     

Hydrocarbon degradation, plant colonization and gene expression of alkane degradation genes by endophytic Enterobacter ludwigii strains

The genus Enterobacter comprises a range of beneficial plant-associated bacteria showing plant growth promotion. Enterobacter ludwigii belongs to the Enterobacter cloacae complex and has been reported to include human pathogens but also plant-associated strains with plant beneficial capacities. To assess the role of Enterobacter endophytes in hydrocarbon degradation, plant colonization, abundance and expression of CYP153 genes in different plant compartments, three plant species (Italian ryegrass, birdsfoot trefoil and alfalfa) were grown in sterile soil spiked with 1% diesel and inoculated with three endophytic E. ludwigii strains. Results showed that all strains were capable of hydrocarbon degradation and efficiently colonized the rhizosphere and plant interior. Two strains, ISI10-3 and BRI10-9, showed highest degradation rates of diesel fuel up to 68% and performed best in combination with Italian ryegrass and alfalfa. All strains expressed the CYP153 gene in all plant compartments, indicating an active role in degradation of diesel in association with plants.     

Microbial activity during composting of anthracene-contaminated soil

Microbial activity of an anthracene-spiked soil mixed with kitchen waste during laboratory composting at 56-59 degrees C was studied using an in-vessel technology. The effect of old compost containing acclimated microorganisms on the composting efficiency was also investigated. Microbial succession, microbial enzyme activity, microbial diversity and anthracene removal rate were analyzed during 42 days of composting. The results demonstrated that inoculating with old compost increased the amounts of thermophilic microorganisms, but did not significantly increase anthracene removal. A microbial succession from mesophilic bacteria to thermophilic bacteria and thermophilic actinomycetes was observed during composting. Polyphenol oxidase activity decreased while catalase activity varied irregularly. Microbial diversity increased drastically when temperature elevated from 35 to 56 degrees C, but decreased when temperature maintained at 56-59 degrees C.     

Bioremediation and reclamation of soil contaminated with petroleum oil hydrocarbons by exogenously seeded bacterial consortium: a pilot-scale study

Purpose  

Spillage of petroleum hydrocarbons causes significant environmental pollution. Bioremediation is an effective process to remediate petroleum oil contaminant from the ecosystem. The aim of the present study was to reclaim a petroleum oil-contaminated soil which was unsuitable for the cultivation of crop plants by using petroleum oil hydrocarbon-degrading microbial consortium.     

菌源对多环芳烃降解菌的筛选及降解性能的影响

高效降解菌的筛选对利用生物修复技术有效去除环境中的多环芳烃具有重要意义。分别以石油污染土壤和焦化废水活性污泥为菌源,分离出芘降解菌和混合PAHs(菲、荧蒽和芘)降解菌共14株并对其降解性能进行对比研究。结果表明,筛选得到的菌株分别属于9个菌属,其中2种菌源共有的菌属为Mycobacterium sp.、Ralstonia sp.和Shinella sp.。芘和PAHs的高效降解菌(CP16和CM32)均属于分支杆菌属(Mycobacterium),来源于焦化废水活性污泥;菌株CP16对芘(50mg/L)的7 d降解率为74.99%,CM32对PAHs(菲50 mg/L、荧蒽和芘各10 mg/L)的7 d降解率为100%。因此,以焦化废水活性污泥为菌源更有利于获得高效的多环芳烃降解菌。     

土壤石油烃污染的植物毒性及植物-微生物联合降解

通过盆栽实验研究了土壤石油烃污染对玉米和水稻根伸长的影响，并在土壤中接种经过筛选得到的石油烃降解菌，研究石油烃降解菌对石油烃毒性的影响以及对土壤中石油烃的降解。研究结果表明，石油烃浓度低于1 000 mg/kg时对玉米的根系生长有一定的刺激生长作用，随着石油烃浓度的增加，刺激根长生长的作用逐渐降低，研究结果表明，水稻根长受石油烃影响较小。通过对不同处理土壤中石油烃降解的研究结果表明，土壤中种植水稻对石油烃有一定的降解作用，但是不同处理下土壤中的石油烃降解率不同，其中水稻微生物联合处理下土壤中石油烃的降解速率最快，培养期内的降解效率达到53.3%。     

Comparative bioremediation potential of four rhizospheric microbial species against lindane

Four microbial species (Kocuria rhizophila, Microbacterium resistens, Staphylococcus equorum and Staphylococcus cohnii subspecies urealyticus) were isolated from the rhizospheric zone of selected plants growing in a lindane contaminated environment and acclimatized in lindane spiked media (5-100 μg mL⁻¹). The isolated species were inoculated with soil containing 5, 50 and 100 mg kg⁻¹ of lindane and incubated at room temperature. Soil samples were collected periodically to evaluate the microbial dissipation kinetics, dissipation rate, residual lindane concentration and microbial biomass carbon (MBC). There was a marked difference (p < 0.05) in the MBC content and lindane dissipation rate of microbial isolates cultured in three different lindane concentrations. Further, the dissipation rate tended to decrease with increasing lindane concentrations. After 45 d, the residual lindane concentrations in three different spiked soils were reduced to 0%, 41% and 33%, respectively. Among the four species, S. cohnii subspecies urealyticus exhibited maximum dissipation (41.65 mg kg⁻¹) and can be exploited for the in situ remediation of low to medium level lindane contaminated soils.     

Effect of biochar on the fate of volatile petroleum hydrocarbons in an aerobic sandy soil

Biochar addition to soil is currently being investigated as a novel technology to remediate polluted sites. A critical consideration is the impact of biochar on the intrinsic microbial pollutant degradation, in particular at sites polluted with a mixture of readily biodegradable and more persistent organic pollutants. We therefore studied the impact of biochar (2% on dry weight basis) on the fate of volatile petroleum hydrocarbons in an aerobic sandy soil with batch and column studies. The soil-water partitioning coefficient, K(d), was enhanced in the biochar-amended soil up to a factor 36, and petroleum hydrocarbon vapor migration was retarded accordingly. Despite increased sorption, in particular of monoaromatic hydrocarbons, the overall microbial respiration was comparable in the biochar-amended and unamended soil. This was due to more rapid biodegradation of linear, cyclic and branched alkanes in the biochar amended soil. We concluded that the total petroleum hydrocarbon degradation rate was controlled by a factor other than substrate availability and the reduced availability of monoaromatic hydrocarbons in the biochar amended soil led to greater biodegradation of the other petroleum compounds.     

A field trial for an ex-situ bioremediation of a drilling mud-polluted site

The remediation of drilling mud-polluted sites in the Southeast of Mexico is a top priority for Mexican oil industry. The objective of this work was to find a technology to remediate these sites. A field trial was performed by composting in biopiles, where four 1ton soil-biopiles were established, one treatment in triplicate and one unamended biopile. Amended biopiles were added with nutrients to get a C/N/P ratio of 100/3/0.5 plus a bulking agent (straw) at a soil/straw ratio of 97/3. Moisture content was maintained around 30-35%. Results showed that, after 180 d, total petroleum hydrocarbon (TPH) concentrations decreased from 99300+/-23000mgTPHkg(-1) soil to 5500+/-770mgTPHkg(-1) for amended biopiles and to 22900+/-7800mgTPHkg(-1) for unamended biopile. An undisturbed soil control showed no change in TPH concentrations. Gas chromatographic analysis showed residual alkyl dibenzothiophene type compounds. Highest bacterial counts were observed during the first 30 d which correlated with highest TPH removal, whereas fungal count increased at the end of the experimentation period. Results suggested an important role of the straw, nutrient addition and water content in stimulating aerobic microbial activity and thus hydrocarbon removal. This finding opens an opportunity to remediate old polluted sites with recalcitrant and high TPH concentration.     

Use of selected autochthonous soil bacteria to enhanced degradation of hydrocarbons in soil

A mixed population of soil hydrocarbon degrading bacteria was used to accelerate the biodegradation of a petrochemical waste. An aromatic hydrocarbon storage tank bottom was mixed with soil (10% w/w). After a month 43% of the hydrocarbons were degraded in uninoculated and in fertilized soil, while 65% were degraded in inoculated soil. Nutrient supplemented vermiculite seems to be a good possibility to produce effective hydrocarbon degrading inoculants.     

Bioremediation of diesel-contaminated soil with composting

The major objective of this research was to find the appropriate mix ratio of organic amendments for enhancing diesel oil degradation during contaminated soil composting. Sewage sludge or compost was added as an amendment for supplementing organic matter for composting of contaminated soil. The ratios of contaminated soil to organic amendments were 1:0.1, 1:0.3, 1:0.5, and 1:1 as wet weight basis. Target contaminant of this research was diesel oil, which was spiked at 10,000 mg/kg sample on a dry weight basis. The degradation of diesel oil was significantly enhanced by the addition of these organic amendments relative to straight soil. Degradation rates of total petroleum hydrocarbons (TPH) and n-alkanes were the greatest at the ratio of 1:0.5 of contaminated soil to organic amendments on wet weight basis. Preferential degradation of n-alkanes over TPH was observed regardless of the kind and the amount of organic amendments. The first order degradation constant of n-alkanes was about twice TPH degradation constant. Normal alkanes could be divided in two groups (C10-C15 versus C16-C20) based on the first order kinetic constant. Volatilization loss of TPH was only about 2% of initial TPH. Normal alkanes lost by volatilization were mainly by the compounds of C10 to C16. High correlations (r=0.80-0.86) were found among TPH degradation rate, amount of CO2 evolved, and dehydrogenase activity.     

复合微生物菌剂强化堆肥技术研究

采用复合微生物菌剂对生活垃圾的接种堆肥技术进行了实验研究。通过测定堆肥过程中反应器出口O2、CO2与H2S气体浓度及对堆肥样品扫描电镜照片分析，比较了3个接种组与1个对照组中堆料中微生物总数变化、种群结构演替及堆肥腐熟速度。试验结果表明，在原料成分为：生活垃圾／成熟堆肥=80／20，有机物约为60％，初始含水率为55％，初始C／N-30时，对于不同接种量的复合微生物接种系统堆料中分别接种0．2％、0．3％、0．5％(质量百分含量)，与加入0．3％灭活菌的对照组进行对比实验，接种复合微生物菌剂堆肥系统不仅微生物总数高于对照组，而且其种群结构合理，能明显提高堆肥效率，有效控制臭气的产生，提高堆肥腐熟度。     

Sorption and Biodegradation of Vapor-Phase Organic Compounds with Wastewater Sludge and Food Waste Compost

ABSTRACT

To test the possible use of composted food waste and wastewater sludge as biofilters to treat gas-phase volatile organic compounds (VOCs), batch experiments were conducted with an isolated strain that could degrade aromatic compounds under aerobic conditions. A benzene and trichloroethylene (TCE) mixture was used as the gas-phase pollutant in experiments with composted food waste, sludge, and soil. Under aerobic conditions, benzene was degraded as a primary substrate and TCE was degraded cometabolically, with water contents varying from 6 to 60% (volume of water added/volume of solid). Optimal water content for VOC removal was 12% for the soil, 36% for the composted food waste, and 48% for the sludge.

The extent of VOC sorption and biodegradation at the optimal water content was different for each material. With the same initial VOC concentration, more VOCs were removed by sorption onto the composted food waste and the sludge, while less VOCs were biodegraded in comparison with the results using soil. The reason the biodegradation in the soil was greater may be partly attributed to the fact that, due to less sorption, the aqueous-phase concentration of VOCs, which microorganisms could utilize as a carbon source or cometabolize, was higher. We also speculate that the distribution of microorganisms in each medium affects the rate of biodegradation. A large number of microorganisms were attached to the composted food waste and sludge. Mass transfer of VOCs and oxygen to these microorganisms, which appear to have been heterogeneously distributed in clusters, may have been limited, resulting in hindered biodegradation.