Use of tracer tests to evaluate the impact of enhanced-solubilization flushing on in-situ biodegradation

Tracer tests were conducted to evaluate the effect of a complexing sugar flush (CSF) on in-situ biodegradation potential at a site contaminated by jet fuel, solvents, and other organic compounds. Technical-grade hydroxypropyl-beta-cyclodextrin was used during the CSF study, which was conducted in a hydraulically isolated cell emplaced in a surficial aquifer. In-situ biodegradation potential was assessed with the use of tracer tests, which were conducted prior to and immediately following the CSF study. Ethanol, hexanol, and benzoate were used as the biodegradable tracers, while bromide was used as a nonreactive tracer. The results indicate that the biodegradation of benzoate was similar for both tracer tests. Conversely, the biodegradation of ethanol (23% increase) and hexanol (41% increase) was greater for the post-CSF tracer test. In addition, analysis of core samples collected from within the test cell indicates that the population density of aerobic jet-fuel degraders increased in the vicinity of the injection wells during the CSF. These results indicate that the cyclodextrin flush did not deleteriously affect the indigenous microbial community. This study illustrates that tracer tests can be used to evaluate the impact of remediation activities on in-situ biodegradation potential.     

Biodegradation of Crude Oil Contaminating Marine Shorelines and Freshwater Wetlands

This paper is a summary of the various factors influencing weathering of oil after it has been released into the environment from a spill incident. Special emphasis has been placed on biodegradation processes. Results from two field studies conducted in 1994 and 1999 involving bioremediation of an experimental oil spill on a marine sandy shoreline in Delaware and a freshwater wetland on the St. Lawrence River in Quebec, Canada have been presented in the paper.     

In-situ Treatment of Oiled Sediment Shorelines

Experimental oil spill studies were conducted to quantify the effectiveness of selected in-situ shoreline treatment options to accelerate natural oil removal processes on mixed-sediment (sand and pebble) shorelines. At each of three distinct shoreline sites, treatment test plots and control plots were established within a 40-, 80- and 143-m continuous stretch of oiled shoreline. A total of 5500 l of oil was deposited along a 3-m wide swath in the upper intertidal zone at each site. Approximately one week after oiling, a different treatment technique was applied to each plot. The treatment techniques were: sediment relocation (surf washing), mixing (tilling), bioremediation (fertilizer application), and bioremediation combined with mixing. One plot at each site was monitored for natural attenuation. The quantity of oil removed from the plots was measured six times up to 60 days post-treatment and then again one year later. Changes in the physical character of the beach, oil penetration, movement of oil to the subtidal environment, toxicity, and biodegradation were monitored over the 400-day period.The results verified quantitatively that relocation of oiled sediments significantly accelerated the rate of oil removal from the shoreline by more than one year. Microscopic observations and image analyses confirmed that the oil-mineral aggregate formation process was active and was increased by sediment relocation. Oil biodegradation occurred in this arctic environment, both in the oiled sediments and on the fine mineral particles removed from the sediment by natural physical processes. The biodegradation of oil in sediment was significantly stimulated by simple bioremediation protocols. Mixing (by tilling) did not clearly stimulate oil loss and natural recovery in the context of this experimental design. None of the treatment techniques elevated toxicity in the nearshore environment to unacceptable levels, nor did they result in consequential alongshore or nearshore oiling.     

Restoring biochemical activity and bacterial diversity in a trichloroethylene-contaminated soil: the reclamation effect of vermicomposted olive wastes

Background, aim, and scope  In this work, the potential for using olive-mill solid waste as an organic amendment for biochemical and biological restoration of a trichloroethylene-contaminated soil, which has previously been stabilized through vermicomposting processes, has been explored. Materials and methods  Trichloroethylene-contaminated water was pumped into soil columns with a layer of vermicompost at 10-cm depth (biobarrier system). The impacts of the trichloroethylene on the microbial community were evaluated by determining: (1) the overall microbial activity (estimated as dehydrogenase activity) and enzyme activities related to the main nutrient cycles (β-glucosidase, o-diphenoloxidase, phosphatase, urease, and arylsulphatase activities). In addition, isoelectric focusing of the soil extracellular humic-β-glucosidase complexes was performed to study the enzymatically active humic matter related to the soil carbon cycle. (2) The soil bacterial diversity and the molecular mechanisms for the bacterial resistance to organic solvents were also determined. For this, polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis (DGGE) was used to detect changes in bacterial community structure and PCR-single-strand conformational polymorphism (SSCP) was developed and optimised for detection and discrimination of the resistance-nodulation-division (RND) genes amplified from the contaminated soils. Results  Vermicompost reduced, with respect to the unamended soil, about 30% of the trichloroethylene leaching during the first month of the experiment. Trichloroethylene had a marked negative effect on soil dehydrogenase, β-glucosidase, urease, phosphatase, and arylsulphatase activities. Nevertheless, the vermicompost tended to avoid this toxic effect. Vermicompost also displays stable humic-β-glucosidase complexes that increased the extracellular activity related to C-cycle in the contaminated soils. The isoelectric focusing technique showed a more biochemically active humic matter in the soil sampled under the vermicompost. The behaviour of the three main phyla of bacteria isolated from the DGGE bands was quite different. Bands corresponding to Actinobacteria disappeared, whereas those affiliated with Proteobacteria remained after the trichloroethylene contamination. The disappeared Actinobacteria became visible in the soil amended with the vermicompost. Bands corresponding to Bacteriodetes appeared only in columns of contaminated soils. In this study, six types of RND proteins were detected by PCR-SSCP in the natural soil, three in the trichloroethylene-contaminated soil and 7/5 in trichloroethylene-contaminated soil above/below the vermicompost in the biobarrier columns. Trichloroethylene tended to reduce or eliminate all the clones detected in the uncontaminated soil, whereas new efflux pumps appeared in the biobarrier columns. Discussion  Although enzymes incorporated into the humic substances of vermicomposted olive wastes are quite stable, trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity in the amended soils. The decrease was less severe in the biobarrier system, but in any case, no relation was found between the levels of trichloroethylene in soil and extracellular β-glucosidase activity, or between the latter and the quantity of humic carbon in soils. The isoelectric focusing technique was carried out in the humic fraction to determine whether the loss of activity occurred in overall extracellular β-glucosidase or in that linked to stable humic substances (humic–enzyme complexes). The contaminated soils showed the lower enzyme activities, whereas contaminated and amended soils presented greater quantity of focalised (and therefore stable) humic carbon and spectra heterogeneity: very different bands with higher enzyme activities. No clear relationship between trichloroethylene concentration in soil and diversity of the bacterial population was noted. Similar patterns could be found when the community structures of bacteria and microbial activity were considered. Since the use of the dehydrogenase assay has been recognised as a useful indicator of the overall measure of the intensity of microbial metabolism, these results could be attributed to PCR-DGGE methodology, since the method reveals the presence of dominant populations regardless of their metabolic state. Trichloroethylene maintained or even increased the number of clones with the DNA encoding for RND proteins, except for the contaminated soil located above the vermicompost. However, the main effect of trichloroethylene was to modify the structure of the community in contaminated soils, considering the type of efflux pumps encoded by the DNA extracted from soil bacteria. Conclusions  Trichloroethylene inhibited specific functions in soil and had a clear influence on the structure of the autochthonous bacterial community. The organic matter released by the vermicomposted olive waste tended to avoid the toxic effect of the contaminant. Trichloroethylene also inhibited the background levels of the soil extracellular β-glucosidase activity, even when vermicompost was present. In this case, the effect of the vermicompost was to provide and/or to stimulate the humic-β-glucosidase complexes located in the soil humic fraction >10⁴, increasing the resistance of the enzyme to the inhibition. The bacterial community from the soil presented significantly different mechanisms to resistance to solvents (RND proteins) under trichloroethylene conditions. The effect of the vermicompost was to induce these mechanisms in the autochthonous bacterial community and/or incorporated new bacterial species, able to grow in a trichloroethylene-contaminated ambient. Coupled biochemical and molecular methodologies are therefore helpful approaches in assessing the effect of an organic amendment on the biochemical and biological restoration of a trichloroethylene-contaminated soil. Recommendations and perspectives  Since the main biochemical and biological effects of the organic amendment on the contaminated soil seem to be the incorporation of biochemically active humic matter, as well as new bacterial species able to grow in a trichloroethylene-contaminated ambient, isoelectric focusing and PCR-SSCP methodologies should be considered as parts of an integrated approach to determine the success of a restoration scheme.     

Improved VOC bioremediation using a fluidized bed peat bioreactor

A gas–solid fluidized bed bioreactor was successfully used to treat air contaminated with a volatile organic compound (VOC). A bioreactor containing both a fluidized and packed bed of moist peat granules removed ethanol, a representative VOC, from an air stream. The fluidized bed operation mode of the bioreactor outperformed the packed bed mode. The maximum elimination capacity (EC) of ethanol in the fluidized mode was 1520 g m⁻³ h⁻¹, with removal efficiencies ranging between 45 and 100%, at loadings up to 3400 g m⁻³ h⁻¹. Maximum EC was 530 g m⁻³ h⁻¹ in the packed bed mode. Removal efficiency in the fluidized bioreactor was best at the lowest velocity, where the bubbling bed fluidization regime predominated. As gas velocity increased, the size and amount of large bubbles (slugs) increased and removal efficiency decreased while elimination capacity increased.     

Uranium removal from groundwater via in situ biostimulation: Field-scale modeling of transport and biological processes

During 2002 and 2003, bioremediation experiments in the unconfined aquifer of the Old Rifle UMTRA field site in western Colorado provided evidence for the immobilization of hexavalent uranium in groundwater by iron-reducing Geobacter sp. stimulated by acetate amendment. As the bioavailable Fe(III) terminal electron acceptor was depleted in the zone just downgradient of the acetate injection gallery, sulfate-reducing organisms came to dominate the microbial community. In the present study, we use multicomponent reactive transport modeling to analyze data from the 2002 field experiment to identify the dominant transport and biological processes controlling uranium mobility during biostimulation, and determine field-scale parameters for these modeled processes. The coupled process simulation approach was able to establish a quantitative characterization of the principal flow, transport, and reaction processes based on the 2002 field experiment, that could be applied without modification to describe the 2003 field experiment. Insights gained from this analysis include field-scale estimates of the bioavailable Fe(III) mineral threshold for the onset of sulfate reduction, and rates for the Fe(III), U(VI), and sulfate terminal electron accepting processes.     

生物修复技术治理农村面源水污染问题的调查研究

调查得知，随着农村化肥、农药、除草剂的大量使用以及禽畜养殖规模化的扩大，造成我国农村面源污染日趋严重。农村面源污染源分散，没有明确位置，其治理一直是个难题。本文结合成都市政府解决府南河水污染问题时采用的针对面源污染的生物修复技术，对此技术作了较深入的研究，并取得了一定的效果，对国内城市近郊农村面源污染有较好的参考价值。在对所需植物进行逐级实验后，可推广至农村以解决成都地区的面源污染问题。     

Earthworm assisted bioremediation of organic contaminants

Due to their biological, chemical and physical actions, earthworms can be directly employed within bioremediation strategies to promote biodegradation of organic contaminants. Earthworms have been shown to aerate and bioturbate soils and improve their nutritional status and fertility, which are variables known to limit bioremediation. Earthworms have also been shown to retard the binding of organic contaminants to soils, release previously soil-bound contaminants for subsequent degradation, and promote and disperse organic contaminant degrading microorganisms. This review discusses these earthworm actions upon the soil environment and how they might influence the fate and behaviour of soil associated organic contaminants, subsequently improving bioremediation potential. The latter part of this review considers organic compounds in the following order: agrochemicals, petroleum and crude oil hydrocarbons, PAHs and PCBs.     

黑麦草-菌根-蚯蚓对多氯联苯污染土壤的联合修复效应

采用水稻土盆栽试验,研究了黑麦草-蚯蚓-菌根对多氯联苯(PCB)污染土壤的联合修复效应.试验土壤中PCB含量为307.2 ng·g-1,并设添加蚯蚓(E)、种植黑麦草(R)、黑麦草-蚯蚓(RE)、黑麦草-接种菌根(RM)及黑麦草-菌根-蚯蚓三者联合修复(RME)5个处理,以不加黑麦草、蚯蚓、菌根的土壤为对照(CK),并...     

Effects of pH and cyclodextrins on pentachlorophenol degradation (mineralization) by white-rot fungi

White-rot fungi (WRF) such as Trametes hirsuta completely degrade (mineralize) pentachlorophenol (PCP) and many other organopollutants. This has led to them being used to decontaminate various substrates (e.g. soil) through biorememediation. However, because PCP is a biocide, it can inhibit fungal growth and thereby its own degradation. It was hypothesized that substrate pH might affect PCP degradation, because when the pH is lower than 4.7 (the pKa for PCP) the phenol predominates, while at higher pH the phenate does. These two PCP species differ markedly in physical and biological properties. The effect of cyclodextrins was also investigated since it is known that the inclusion complexes these form with PCP differ in bioavailability and toxicity from non-complexed PCP. Tests were first made in liquid and agar media (where conditions are relatively easy to control), and then in a sawdust, because it is a common target matrix for WRF bioremediation. Results with the liquid and agar media showed that growth in the presence of inhibitory PCP concentrations decreased as the pH decreased, consistent with the phenol being more toxic. Growth in sawdust was less affected by PCP regardless of the pH, presumably because the PCP sorbed to the wood which decreased its bioavailability. Some cyclodextrins markedly decreased the PCP's toxicity in liquid, agar and sawdust media. Rates of PCP mineralization (measured from production of (14)CO(2) from (14)C PCP) in liquid cultures containing 0.5 mgkg(-1) PCP (a sub-inhibitory concentration) were similar from pH 2.5-7.5, indicating that the phenol and the phenate were equally degradable. Degradation of a growth inhibiting concentration on sawdust (1,000 mgkg(-1)) could be increased slightly by lowering the pH below the pKa, this increasing sorption to the wood. Degradation increased more when the pH was raised well above the pKa, presumably due to the phenate being less toxic and more soluble, making it more available to the degradative system. Although some cyclodextrins decreased growth inhibition, they also interfered with degradation. If this interference could be overcome, cyclodextrins could be used to increase the maximal PCP concentration that could be treated by WRF bioremediation.