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Phytoremediation--a novel and promising approach for environmental clean-up.

Phytoremediation is an eco friendly approach for remediation of contaminated soil and water using plants. Phytoremediation is comprised of two components, one by the root colonizing microbes and the other by plants themselves, which degrade the toxic compounds to further non-toxic metabolites. Various compounds, viz. organic compounds, xenobiotics, pesticides and heavy metals, are among the contaminants that can be effectively remediated by plants. Plant cell cultures, hairy roots and algae have been studied for their ability to degrade a number of contaminants. They exhibit various enzymatic activities for degradation of xenobiotics, viz. dehalogenation, denitrification leading to breakdown of complex compounds to simple and non-toxic products. Plants and algae also have the ability to hyper accumulate various heavy metals by the action of phytochelatins and metallothioneins forming complexes with heavy metals and translocate them into vacuoles. Molecular cloning and expression of heavy metal accumulator genes and xenobiotic degrading enzyme coding genes resulted in enhanced remediation rates, which will be helpful in making the process for large-scale application to remediate vast areas of contaminated soils. A few companies worldwide are also working on this aspect of bioremediation, mainly by transgenic plants to replace expensive physical or chemical remediation techniques. Selection and testing multiple hyperaccumulator plants, protein engineering ofphytochelatin and membrane transporter genes and their expression would enhance the rate of phytoremediation, making this process a successful one for bioremediation of environmental contamination. Recent years have seen major investments in the R&D, which have also resulted in competition of filing patents by several companies for economic gains. The details of science & technology related to phytoremediation have been discussed with a focus on future trends and prospects of global relevance.

Biodegradation, Environmental↗

Aspergilli and lignocellulosics: enzymology and biotechnological applications.

Aspergilli are versatile ascomycetes that are able to transform at a rapid rate a wide spectrum of lignin-related aromatic compounds. While it is clear that these fungi can degrade phenolic and polysaccharide components from lignocellulosic material, the status regarding degradation of high-molecular mass lignins is controversial. This review compiles data from the literature as well as that from the authors' laboratory with the aim of clarifying this point. The main body of evidence points towards the inability of aspergilli alone to degrade lignin free of low-molecular mass contaminants. Nevertheless, the ability of this genus to efficiently degrade hemicelluloses makes it an essential participant in the complex microbial system necessary for wood decay under natural conditions. Aspergilli are known to overproduce high levels of hemicellulolytic enzymes. Out of the large array of these enzymes that act in concert to degrade lignocellulosic material, only endoxylanases of aspergilli are described in so far as these are the main activities required for enzyme-aided bleaching. The biochemical features of the endoxylanases from Aspergillus niger are briefly described as these serve to illustrate how a complex family of isozymes is necessary to deal with the structural and chemical heterogeneity of xylans. Emphasis is placed on the biotechnological applications of lignocellulosic materials transformed by aspergilli. The key application areas are biopulping and biobleaching where a reduction in the use of environmentally harmful chemicals traditionally used in the pulp and paper industry is envisaged. Waste water treatment represents another vast application area where aspergilli have been shown to be effective not only in colour removal but also in the bioconversion of potentially noxious substances into useful bioproducts.

Aspergillus↗

A mathematical model for the transport and fate of organic chemicals in unsaturated/saturated soils.

A mathematical model, simulating the transport and fate of nonionizable organic compounds in unsaturated/saturated porous media (soils) in a terrestrial microcosm has been developed. Using the principles of water mass, momentum, heat energy and chemical mass balance, the three fields: moisture, temperature, and liquid phase chemical concentration are solved for simultaneously by coupling the soil slab to an environmentally realistic air-soil interface (a dynamic free boundary) conditions and a prescribed height water table. The environmental conditions at the soil surface-air chamber interface are easily changed, via geometric scaling factors, to simulate either an open agricultural field or a landfill type of situation. Illustrative simulation runs examine the effects of different soil-chemical characteristics on hydrological and chemical concentration profiles.

Biodegradation, Environmental↗

Phosphorus removal from Everglades agricultural area runoff by submerged aquatic vegetation/limerock treatment technology: an overview of research.

The 1994 Everglades Forever Act mandates the South Florida Water Management District and the Florida Department of Environmental Protection to evaluate a series of advanced treatment technologies to reduce total phosphorus (TP) in Everglades Agricultural Area runoff to a threshold target level. A submerged aquatic vegetation/limerock (SAV/LR) treatment system is one of the technologies selected for evaluation. The research program consists of two phases. Phase I examined the efficiency of SAV/LR treatment system for TP removal at the mesocosm scale. Preliminary results demonstrate that this technology is capable of reducing effluent TP to as low as 10 microg/L under constant flows. The SAV component removes the majority of the influent soluble reactive P, while the limerock component removes a portion of the particulate P. Phase II is a multi-scale project (i.e., microcosms, mesocosms, test cells and full-size wetlands). Experiments and field investigations using various environmental scenarios are designed to (1) identify key P removal processes; (2) provide management and operational criteria for basin-scale implementation; and (3) provide scientific data for a standardized comparison of performance among advanced treatment technologies.

Agriculture↗

Evaluation of manometric respiration tests to assess the effects of veterinary antibiotics in soil.

Extensive use of antimicrobials in veterinary medicine results in environmental exposure. Of major concern are microbial effects; including effects on nutrient soil cycles and antibiotic resistance. There is a need to assess the effects of these compounds in the environment. The application of standardized guidelines is relevant in studying many compounds. However there is a lack of special test methods designed for antibiotics. We validate manometric test flasks using glucose and a recalcitrant herbicide. The suitability of these tests for studying antibacterial agents is then investigated using two target functions (aerobic biodegradation and carbon transformation). Compound stability is quantified using HPLC techniques. Effects on total soil respiration in the biodegradation test are immediate and differ significantly from background. We show that compounds do not function as substrates, so effects are due to other soil processes, correlate well to sorption characteristics and are not dose dependent. This test provides details of relative antimicrobial potency towards soil microorganisms and can be used to rank compounds. However the test does not provide details on the nature or extent of specific microbial effects. In contrast, the carbon transformation test is more specific and provides a reproducible indication of dose effect relationships, which is more suitable in assessing the effects of these compounds in the environment. Presently, standard guidelines do not take into account the normal input of antibiotics into soils via contaminated sludge or manure. This should be corrected in future guidelines as these inputs alter microbial composition, organic matter, ionic strength and pH affect sorption and overall impact the test results.

Anti-Bacterial Agents↗

The influence of EDDS on the uptake of heavy metals in hydroponically grown sunflowers.

Phytoextraction is an environmentally friendly in situ technique for cleaning up metal contaminated land. Unfortunately, efficient metal uptake by remediation plants is often limited by low phytoavailability of the targeted metals. Chelant assisted phytoextraction has been proposed to improve the efficiency of phytoextraction. Phytoremediation involves several subsequent steps: transfer of metals from the bulk soil to the root surfaces, uptake into the roots and translocation to the shoots. Nutrient solution experiments address the latter two steps. In this context we investigated the influence of the biodegradable chelating agent SS-EDDS on uptake of essential (Cu and Zn) and non-essential (Pb) metals by sunflowers from nutrient solution. EDDS was detected in shoots and xylem sap for the first time, proving that it is taken up into the above ground biomass of plants. The essential metals Cu and Zn were decreased in shoots in the presence of EDDS whereas uptake of the non-essential Pb was enhanced. We suggest that in the presence of EDDS all three metals were taken up by the non-selective apoplastic pathway as the EDDS complexes, whereas in the absence of EDDS essential metal uptake was primarily selective along the symplastic pathway. This shows that synthetic chelating agents do not necessarily increase uptake of heavy metals, when soluble concentrations are equal in the presence and absence of chelates.

Biodegradation, Environmental↗

Bioremedial potential of microbial mechanisms of metal mobilization and immobilization.

Microorganisms play important roles in the environmental fate of toxic metals and radionuclides with a multiplicity of mechanisms effecting transformations between soluble and insoluble forms. These mechanisms are integral components of natural biogeochemical cycles and are of potential for both in situ and ex situ bioremedial treatment processes for solid and liquid wastes.

Alkylation↗

Determination of chemical warfare agent degradation products at low-part-per-billion levels in aqueous samples and sub-part-per-million levels in soils using capillary electrophoresis.

A significant enhancement in the method detection limits is observed in the analysis of chemical warfare agent (CWA) degradation products in environmental samples by capillary electrophoresis (CE) using electrokinetic injection. The CE method uses indirect UV detection of the nonderivitized acidic analyte and a cationic surfactant, didodecyldimethylammonium hydroxide, for reversal of the electroosmotic flow. Analytes studied include the dibasic acid methylphosphonic acid (MPA) and its monoacid/monoalkyl esters, RMPA, where R = ethyl, isopropyl, and pinacolyl (2-(3,3-dimethylbutyl)). The CE method uses an attractive buffer system which is highly stable and inexpensive, and, in addition to reversing the electroosmotic flow, provides excellent separation efficiencies within a 3-min run. This CE method is also free from interference caused by carbonate, humic acids, and fluoride. Compared to pressure injection, electrokinetic injection with this CE buffer system provided substantially lower detection limits, up to 100-fold lower for samples in reagent water. However, to best realize the benefits of the electrokinetic injection enhancement for environmental samples, a prior cleanup of the sample using standard ion-exchange cartridges is necessary. This cleanup step uses sequential cartridges to remove sulfate (barium cartridge), chloride (silver cartridge), and cations (H+ cartridge). Using this approach, detection limits for these four acids were as low as 1-2 micrograms/L for water samples and 25-50 micrograms/L for aqueous leachates of soil samples (10 mL of leachate/1.5 g of soil). The utility of this method for separation of CWA degradation products by CE is discussed in terms of pressure injection versus electokinetic injection. The effects of voltage and time of injection on the separation were investigated. Results from three types of soils and four types of water (groundwater, artificial seawater, tap water, bay water) indicated that the method has potential for environmental monitoring. Quantitative CE analysis with electrokinetic injection enhancement of detection limits of these types of environmental samples requires the use of an appropriate internal standard approach. The data presented here indicate that an internal standard-based approach could be expected to give analysis results in the sub-part-per-million concentration range of 90-110% of the true value.

Biodegradation, Environmental↗

Assessing the role of alkane hydroxylase genotypes in environmental samples by competitive PCR.

AIMS: A molecular tool for extensive detection of prokaryotic alkane hydroxylase genes (alkB) was developed. AlkB genotypes involved in the degradation of short-chain alkanes were quantified in environmental samples in order to assess their occurrence and ecological importance. METHODS AND RESULTS: Four primer pairs specific for distinct clusters of alkane hydroxylase genes were designed, allowing amplification of alkB-related genes from all tested alkane-degrading strains and from six of seven microcosms. For the primer pair detecting alkB genes related to the Pseudomonas putida GPo1 alkB gene and the one targeting alkB genes of Gram-positive strains, both involved in short-chain alkane degradation (<C10), a quantitative competitive PCR (cPCR) assay was developed and validated on alkB-containing strains. AlkB genes of the two groups were then quantified in hydrocarbon-contaminated and pristine freshwater and soil samples, and their respective frequency was compared to degradation rates of short-chain n-alkanes. Pseudomonas putida-related alkB genes were prevalent in freshwater samples, but Gram-positive alkB-containing strains were more consistently related to alkane degradation activities. The latter genotype was more abundant in soils, although both genotypes increased in the most contaminated soils studied. CONCLUSIONS: Predominance of alkB genotypes depends on the ecosystem and environmental conditions, but alkane exposure generally leads to an increase of both studied genotypes. SIGNIFICANCE AND IMPACT OF THE STUDY: The study illustrates the distribution of two different alkB genotypes in two types of ecosystems, and highlights their respective roles in the environment.

Alkanes↗

Uranium reduction.

The dramatic decrease in solubility accompanying the reduction of U(VI) to U(IV), producing the insoluble mineral uraninite, has been viewed as a potential mechanism for sequestration of environmental uranium contamination. In the past 15 years, it has been firmly established that a variety of bacteria exhibit this reductive capacity. To obtain an understanding of the microbial metal metabolism, to develop a practical approach for the acceleration of in situ bioreduction, and to predict the long-term fate of environmental uranium, several aspects of the microbial process have been experimentally explored. This review briefly addresses the research to identify specific uranium reductases and their cellular location, competition between uranium and other electron acceptors, attempts to stimulate in situ reduction, and mechanisms of reoxidation of reduced uranium minerals.

Bacteria↗

Heterologous expression of a mammalian ABC transporter in plant and its application to phytoremediation.

Mammalian ATP-binding cassette (ABC) transporters involved in the multidrug-resistance of cancer cells can efflux cytotoxic compounds that show a wide variety of chemical structures and biological activities. Human multidrug resistance-associated protein (hMRP1) is one of the most intensively studied ABC transporters and many substrates have been identified, including both organic and inorganic compounds. In an attempt at novel 'transport engineering' using hMRP1 as a molecular pump, we established transgenic tobacco plants that showed clear resistance to cadmium and daunorubicin, although they were not resistant to etoposide, another known substrate of hMRP1. When expressed in tobacco cells, hMRP1 protein was localized at vacuolar membrane, while members of the MRP family are localized at plasma membrane in mammalian cells to reduce the cellular accumulation of various drugs. Thus, the hMRP1-expressing tobacco cells were able to take up these substrates across the tonoplast and sequestrate them in the vacuolar matrix. These results suggest that it may be possible to use the transgenic tobacco in phytoremediation, where a single transformation with an ABC transporter with broad substrate specificity should be effective for extracting various environmental pollutants including both organic and inorganic compounds, and accumulate them in the plant body. This should be advantageous for the remediation of a complex polluted environment, which is commonly found in the real world.

Antibiotics, Antineoplastic↗

Environmental risk assessment of acrylamide and methylolacrylamide from a grouting agent used in the tunnel construction of Romeriksporten, Norway.

Increased focus on the possible evironmental risk associated with large-scale use of grouting agents has revealed that leakage of chemicals from grouting activities may cause harm to the environment. Chemical grouting agents are used to reduce water leakages in e.g. tunnel constructions. The present study focuses on monitoring results and environmental risk caused by releases of non-polymerised monomers during use of the acrylamide-based grouting agent Rhoca-Gil (Siprogel). Drainage water from the tunnel Romeriksporten was monitored with respect to acrylamide and methylolacrylamide, as leakage of these substances was observed earlier in connection with similar constructions where Rhoca-Gil was used. Concentrations of acrylamide and methylolacrylamide in the draining water showed that these substances leaked out both in connection with the injection of Rhoca-Gil and in connection with after-injection using other grouting agents. Gel formation studies with Rhoca-Gil showed that a low degree of polymerisation (and hence, large leakages of monomers) can be expected if the product is diluted with water. Results from investigation of the environmental fate of methylolacrylamide showed that this substance is chemically transformed at the high-pH conditions of the tunnel. Ecotoxicological testing of the substances and an environmental effects assessment for the receiving water (River Alna and the Oslofjord) showed that the discharge of acrylamide and methylolacrylamide may have caused effects on the aquatic life in the river and in a limited area of the fjord.

Acrylamide↗

Dissolving the problem of infectious medical waste.

Decisions in purchasing healthcare products are based on fiscal responsibility, employee safety, regulatory agency guidelines, and environmental conservation. One of today's major dilemmas in health care is that of disposing of medical waste. The Isolyser Company of Norcross, Georgia, is one of the first to develop a biocycle system to reduce the problem of contaminated waste. A description of the system is provided, along with its advantages and disadvantages.

Biodegradation, Environmental↗

Is bioaugmentation a feasible strategy for pollutant removal and site remediation?

Microorganisms can degrade numerous organic pollutants owing to their metabolic machinery and to their capacity to adapt to inhospitable environments. Thus, microorganisms are major players in site remediation. However, their efficiency depends on many factors, including the chemical nature and the concentration of pollutants, their availability to microorganisms, and the physicochemical characteristics of the environment. The capacity of a microbial population to degrade pollutants within an environmental matrix (e.g. soil, sediment, sludge or wastewater) can be enhanced either by stimulation of the indigenous microorganisms by addition of nutrients or electron acceptors (biostimulation) or by the introduction of specific microorganisms to the local population (bioaugmentation). Although it has been practiced in agriculture and in wastewater treatment for years, bioaugmentation is still experimental. Many factors (e.g. predation, competition or sorption) conspire against it. However, several strategies are currently being explored to make bioaugmentation a successful technology in sites that lack significant populations of biodegrading microorganisms. Under optimal local conditions, the rate of pollutant degradation might increase upon addition of an inoculant to remediate a chemical spill; however, the most successful cases of bioaugmentation occur in confined systems, such as bioreactors in which the conditions can be controlled to favour survival and prolonged activity of the exogenous microbial population.

Biodegradation, Environmental↗

Environmental health aspects of fungicides. I. Dithiocarbamtes.

A variety of dithiocarbamate fungicides, as well as their principal trace impurities, and metabolic or chemical degradation products are reviewed. Major focus is on their chemical and toxicological features (primarily carcinogenic, teratogenic, and mutagenic), use patterns, and residues. The compounds discussed include (1) ethylene bis(dithiocarbamated) (maneb, zineb, nabam), (2) dithiocarbamates (febram, ziram), and ("3) thiuram disulfate (thiram) and ethylene thiourea.

Animals↗

Synergistic effect of Glomus intraradices and Frankia spp. on the growth and stress recovery of Alnus glutinosa in an alkaline anthropogenic sediment.

The presence of actinorhizas and arbuscular mycorrhizas may reduce plant stresses caused by adverse soil conditions. A greenhouse experiment was conducted using a sediment with a high pH, resulting from the disposal of waste originated at an acetylene and polyvinylchloride factory, in which Black alder (Alnus glutinosa) seedlings were inoculated either with Glomus intraradices BEG163 (originally isolated from the same sediment), Frankia spp. or both symbionts. After a 6-month growth period, plants inoculated with both symbionts had significantly greater leaf area, shoot height and total biomass when compared with the uninoculated control, the Frankia spp. and the G. intraradices treatments alone. In dual inoculated plants the N and P leaf content was significantly increased. A defoliation experiment was performed to evaluate the stress recovery of A. glutinosa and plants inoculated with both symbionts had a faster leaf regrowth and produced greater numbers of leaves. The dual inoculation resulted in greater numbers of and larger root nodules than when inoculated with Frankia spp. alone. The length and NADH diaphorase activity of the extraradical mycelium of G. intraradices was also significantly greater when dual inoculation was performed. The inoculation with Frankia spp. alone was shown to improve A. glutinosa growth, whereas G. intraradices alone had no positive effect under these environmental conditions. However, when the two symbionts were inoculated together a synergistic effect was observed resulting in a greater benefit for the plants and for both symbionts. The relevance of these findings for the phytorestoration of anthropogenic stressed sediments with high pH is discussed.

Alnus↗

[Remediation of polycyclic aromatic hydrocarbons in non-fluid medium with immobilized microorganism technique].

The remediation of polycyclic aromatic hydrocarbons (PAHs) in non-fluid medium is a difficult and urgent task for environmental scientists. Due to the particularities of non-fluid medium, conventional techniques cannot work effectively, while bioremediation techniques using free microorganism also have many limitations that restricted the use of microorganism in removing PAHs from non-fluid medium. Immobilized microorganism (IM) technique is a proven approach in increasing the stability of microorganism under unfavorable conditions, which can not only provide a special microenvironment to protect dominant microorganisms from the malignant competition of aboriginal microorganism, but also improve the tolerance against toxic compounds in environment. Biodegradation using immobilized cells has been widely investigated for numerous toxic compounds in fluid medium such as in wastewater, or in half-fluid medium such as in slurry. In this paper, based on analyzing the characteristics of IM technique and its feasibility, the problem and development of future research were presented, and the new technique using IM to remediate PAHs in non-fluid medium was put forward.

Bacteria↗

The University of Minnesota Biocatalysis/Biodegradation Database: specialized metabolism for functional genomics.

The University of Minnesota Biocatalysis/Biodegradation Database (UM-BBD, http://www.labmed.umn.edu/umbbd/i nde x.html) first became available on the web in 1995 to provide information on microbial biocatalytic reactions of, and biodegradation pathways for, organic chemical compounds, especially those produced by man. Its goal is to become a representative database of biodegradation, spanning the diversity of known microbial metabolic routes, organic functional groups, and environmental conditions under which biodegradation occurs. The database can be used to enhance understanding of basic biochemistry, biocatalysis leading to speciality chemical manufacture, and biodegradation of environmental pollutants. It is also a resource for functional genomics, since it contains information on enzymes and genes involved in specialized metabolism not found in intermediary metabolism databases, and thus can assist in assigning functions to genes homologous to such less common genes. With information on >400 reactions and compounds, it is poised to become a resource for prediction of microbial biodegradation pathways for compounds it does not contain, a process complementary to predicting the functions of new classes of microbial genes.

Bacteria↗