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Effect of selected environmental factors on degradation and mineralization of biaryl compounds by the bacterium Ralstonia pickettii in soil and compost.

By varying selected environmental factors, the degradation and mineralization of biaryl compounds by the bacterium Ralstonia pickettii in soil and compost were investigated. An optimized soil moisture and enhanced bioavailability by using the nonionic surfactant Tween 80 were of great importance for the degradation rates of biaryl compounds like biphenyl and 4-chlorobiphenyl by cells of Ralstonia picketti SBUG 290 inoculated into soil. Additionally, degradation of these compounds by the investigated strain in soil was strongly dependent upon the medium of precultivation. Also the influence of temperature and soil pH-value was tested. In contrast to the used soil, the autochthonous flora of the compost seemed to have a higher physiological activity. All investigated compounds (biphenyl, 4-chlorobiphenyl and dibenzofuran) were degraded quickly in compost. Inoculation with the investigated bacterium did not enhance the degradation rates significantly.

Air Pollutants, Occupational↗

Degradation of explosives by nitrate ester reductases.

Explosive-contaminated land poses a hazard both to the environment and to human health. Microbial enzymes, either in their native or heterologous hosts, are a powerful and low-cost tool for eliminating this environmental hazard. As many explosives have only been present in the environment for 10 years, and with similar molecules not known in Nature, the origin of enzymes specialized for the breakdown of explosives is of particular interest. Screening of environmental isolates resulted in the discovery of flavoproteins capable of denitrating the explosives pentaerythritol tetranitrate (PETN) and glycerol trinitrate. These nitrate ester reductases are related in sequence and structure to Old Yellow Enzyme from Saccharomyces carlsbergenisis. All the members of this family have alpha/beta barrel structures and FMN as a prosthetic group, and reduce various electrophilic substrates. The nitrate ester reductases are, however, unusual in that they display activity towards the highly recalcitrant, aromatic explosive 2,4,6-trinitrotoluene, via a reductive pathway resulting in nitrogen liberation. We have embarked on a detailed study of the structure and mechanism of PETN reductase from a strain of Enterobacter cloacae. Work is focused currently on relating structure and function within this growing family of enzymes, with a view to engineering novel enzymes exhibiting useful characteristics.

Biodegradation, Environmental↗

Degradation of methylmercury by bacteria isolated from environmental samples.

A total of 207 bacterial cultures, isolated from environmental samples, was screened for ability to degrade methylmercury. Of these, 30 were found positive for aerobic demethylation. Twenty-two of these were shown to be facultative anaerobes and 21 of these degraded methylmercury anaerobically. All positive species volatilized methylmercury aerobically, and methane was produced as a degradation product. Although methylmercury degradation was complete in most cases, material balances indicated some of the inorganic mercury formed was not volatilized and is presumed bound to the cells. All positive isolates were tolerant to at least 0.5 mug of methylmercury per ml, and the extent of volatilization of mercury increased with concentration to the threshold value. The results indicate that demethylating species are prevalent in the environment and may be important in suppressing the methylmercury content of sediments.

Aerobiosis↗

An entropy based method to evaluate hazardous inorganic substance balances of waste treatment systems.

At present, methods to assess the environmental performance of waste treatment systems are based mainly on emission standards. Since waste treatment yields also products used for recycling and/or land filling, new additional instruments are needed to analyse and evaluate the partitioning of substances during waste treatment. It is shown that in order to fulfil the goals of European Waste Management, waste treatment systems have to concentrate hazardous inorganic substances. Based on substance flow analysis and the statistical entropy function, a method is presented to classify waste treatment systems in view of their power to concentrate or dilute substances. The method allows for the determination of the Substance Concentrating Efficiency (SCE) of waste treatment systems quantitatively. The SCE approach is applied in a case study comparing waste incineration and mechanical-biological waste processing. The results show that this new tool is useful to support decisions regarding the selection of waste treatment systems.

Biodegradation, Environmental↗

Applications of the anaerobic digestion process.

At the start of the new millennium waste management has become a political priority in many countries. One of the main problems today is to cope with an increasing amount of primary waste in an environmentally acceptable way. Biowastes, i.e., municipal, agricultural or industrial organic waste, as well as contaminated soils etc., have traditionally been deposited in landfills or even dumped into the sea or lakes without much environmental concern. In recent times, environmental standards of waste incineration and controlled land filling have gradually improved, and new methods of waste sorting and resource/energy recovery have been developed. Treatment of biowastes by anaerobic digestion processes is in many cases the optimal way to convert organic waste into useful products such as energy (in the form of biogas) and a fertilizer product. Other waste management options, such as land filling and incineration of organic waste has become less desirable, and legislation, both in Europe and elsewhere, tends to favor biological treatment as a way of recycling minerals and nutrients of organic wastes from society back to the food production and supply chain. Removing the relatively wet organic waste from the general waste streams also results in a better calorific value of the remainder for incineration, and a more stable fraction for land filling.

Anaerobiosis↗

The identification of thresholds of acceptability and danger: the chemical presence route.

European Union chemical legislation requires the calculation of local and regional Predicted Environmental Concentrations (PEC) for the assessment of the exposure of new and existing chemicals to aquatic and terrestrial ecosystems. Current methods use local models for air, water and soil to estimate chemical concentrations close to the source and a generic multimedia 'unit world' approach to estimate regional PECs. These models assume generic environmental scenarios representing typical situations in European countries and do not account for the spatial heterogeneity and temporal variability in ecosystem characteristics, soil properties, river flow rates, chemical emissions, etc. The environmental and ecological complexity can best be represented in a Geographic Information System (GIS). By coupling a GIS with a fate simulation model the concentrations of substances in a specific environment are predicted more realistically. The GREAT-ER project (Geography-referenced Regional Exposure Assessment Tool for European Rivers) was launched to refine regional and local exposure assessments for down-the-drain chemicals by applying real, spatial-referenced datasets instead of generic or average values. A modular approach was developed consisting of a hydrological model and a waste-flow, river quality and fate model which are linked to a regional GIS-database. For the calibration and validation in two European study areas representative detergent chemicals (LAS, boron) are used. In a parallel study, high-volume intermediates discharged into the river Rhine are simulated.

Alkanesulfonates↗

Quantification of in situ nutrient and heavy metal remediation by a small pearl oyster (Pinctada imbricata) farm at Port Stephens, Australia.

The use of pearl oysters has recently been proposed as an environmental remediation tool in coastal ecosystems. This study quantified the nitrogen, phosphorus and heavy metal content of the tissue and shell of pearl oysters harvested from a small pearl oyster farm at Port Stephens, Australia. Each tonne of pearl oyster material harvested resulted in approximately 703 g metals, 7452 g nitrogen, and 545 g phosphorus being removed from the waters of Port Stephens. Increasing current farm production of 9.8 tyr(-1) to 499 tyr(-1) would balance current nitrogen loads entering Port Stephens from a small Sewage Treatment Plant (STP) located on its southern shores. Furthermore, manipulation of harvest dates to coincide with oyster condition would likely remove substantially greater quantities of nutrients. This study demonstrates that pearl aquaculture may be used to assist in the removal of pollutants from coastal waters while producing a commercially profitable commodity.

Animals↗

Biosorption of organochlorine pesticides using fungal biomass.

Cladosporium strain AJR(3)18501 was tested for its ability to sorb the organochlorine pesticide (OCP) p,p'-DDT from aqueous media. When p,p'-DDT was added to distilled water, ethanol or 1-propanol solutions in excess of its solubility, p,p'-DDT was sorbed onto the fungal biomass. Increasing the amount of p,p'-DDT in solution by changing the medium composition increased sorbent uptake: p,p'-DDT uptake by the fungal biomass was 2.5 times greater in 25% 1-propanol (17 mg of p,p'-DDT g(-1) dry weight fungal biomass) than in distilled water. When p,p'-DDT was dissolved in 25% 1-propanol (12 mg x l(-1)), rapid p,p'-DDT sorption occurred during the first 60 min of incubation. p,p'-DDT in solution was reduced to 2.5 mg x l(-1) with the remaining p,p'-DDT recovered from the fungal biomass. A number of environmental parameters were tested to determine their effect on p,p'-DDT biosorption. As arsenic (As) is prevalent at DDT-contaminated cattle dip sites, its effect on p,p'-DDT uptake was determined. The presence of As [As(III) or As(V) up to 50 mg x l(-1)] did not inhibit p,p'-DDT uptake and neither As species could be sorbed by the fungal biomass. Changing the pH of the medium from pH 3 to 10 had a small effect on p,p'-DDT sorption at low pH indicating that an ion exchange process is not the major mechanism for p,p'-DDT sorption. Other mechanisms such as Van der Waals forces, chemical binding, hydrogen bonding or ligand exchange may be involved in p,p'-DDT uptake by Cladosporium strain AJR(3)18501.

1-Propanol↗

Biofiltration of volatile organic compounds.

The removal of volatile organic compounds (VOCs) from contaminated airstreams has become a major air pollution concern. Improvement of the biofiltration process commonly used for the removal of odorous compounds has led to a better control of key parameters, enabling the application of biofiltration to be extended also to the removal of VOCs. Moreover, biofiltration, which is based on the ability of micro-organisms to degrade a large variety of compounds, proves to be economical and environmentally viable. In a biofilter, the waste gas is forced to rise through a layer of packed porous material. Thus, pollutants contained in the gaseous effluent are oxidised or converted into biomass by the action of microorganisms previously fixed on the packing material. The biofiltration process is then based on two principal phenomena: (1) transfer of contaminants from the air to the water phase or support medium, (2) bioconversion of pollutants to biomass, metabolic end-products, or carbon dioxide and water. The diversity of biofiltration mechanisms and their interaction with the microflora mean that the biofilter is defined as a complex and structured ecosystem. As a result, in addition to operating conditions, research into the microbial ecology of biofilters is required in order better to optimise the management of such biological treatment systems.

Aerobiosis↗

Two-dimensional gas chromatography coupled to triple quadrupole mass spectrometry for the unambiguous determination of atropisomeric polychlorinated biphenyls in environmental samples.

Two-dimensional gas chromatography coupled to triple quadrupole mass spectrometry (GC-GC-MS-MS) is presented as a robust, selective and sensitive method for the accurate quantification of enantiomeric fractions (EFs) of atropisomeric polychlorinated biphenyls (PCBs) in a range of environmental matrices. The method was subjected to a set of quality control and validation experiments. EF precision of the atropisomers 95, 149, 132, and 174 ranged from 0.7 to 1.4% in soil samples. Quantified amounts of these congeners in soil and compost samples (59<n<100) were on average within 3-26% of the numbers gathered with conventional (achiral) bench top GC-MS. However, significant deviations were observed between EFs obtained by GC-GC-MS-MS and those by GC-GC-MS, GC-MS-MS, and GC-MS. The method was successfully applied to a variety of sample types, such as soil, air, herring, human milk, and, for the first time, compost. EFs in soils are shown to vary with land use and correlations with soil pH were observed. EFs in compost samples were mostly close to, or racemic.

Biodegradation, Environmental↗

Evaluation and comparison of multimedia mass balance models of chemical fate: application of EUSES and ChemCAN to 68 chemicals in Japan.

The European Union System for Evaluation of Substances (EUSES) and the ChemCAN chemical fate model are applied to describe the fate of 68 chemicals on two spatial scales in Japan. Emission information on the chemicals has been obtained from Japan's Pollutant Release and Transfer Registry and available monitoring data gathered from government reports. Environmental concentrations calculated by the two models for the four primary environmental media of air, water, soil and sediment agree within a factor of 3 for over 70% of the data, and within a factor of 10 for over 87% of the data. Reasons for certain large discrepancies are discussed. Concentrations calculated by the models are generally consistent with the lower range of concentrations that are observed in the environment. Agreement between modeled and observed concentrations is considerably improved by including an estimate of the advective input of chemicals in air from outside Japan. The agreement between the EUSES and ChemCAN models suggests that results of individual chemical assessments are not likely to be significantly affected by the choice of chemical fate model. Primary sources of discrepancy between modeled and observed concentrations are believed to be uncertainties in emission rates, degradation half-lives, and the lack of data on advective inflow of contaminants in air.

Biodegradation, Environmental↗

Ecotoxicological evaluation of in situ bioremediation of soils contaminated by the explosive 2,4,6-trinitrotoluene (TNT).

To evaluate the environmental relevance of in situ bioremediation of contaminated soils, effective and reliable monitoring approaches are of special importance. The presented study was conducted as part of a research project investigating in situ bioremediation of topsoils contaminated by the explosive 2,4,6-trinitrotoluene (TNT). Changes in soil toxicity within different experimental fields at a former ordnance factory were evaluated using a battery of five bioassays (plant growth, Collembola reproduction, soil respiration, luminescent bacteria acute toxicity and mutagenicity test) in combination to chemical contaminant analysis. Resulting data reveal clear differences in sensitivities between methods with the luminescent bacteria assay performed with soil leachates as most sensitive toxicity indicator. Complete test battery results are presented in so-called soil toxicity profiles to visualise and facilitate the interpretation of data. Both biological and chemical monitoring results indicate a reduction of soil toxicity within 17 months of remediation.

Analysis of Variance↗

Influence of environmental factors on 2,4-dichlorophenoxyacetic acid degradation by Pseudomonas cepacia isolated from peat.

A Pseudomonas cepacia, designated strain BRI6001, was isolated from peat by enrichment culture using 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole carbon source. BRI6001 grew at up to 13 mM 2,4-D, and degraded 1 mM 2,4-D at an average starting population density as low as 1.5 cells/ml. Degradation was optimal at acidic pH, but could also be inhibited at low pH, associated with chloride release from the substrate, and the limited buffering capacity of the growth medium. The only metabolite detected during growth on 2,4-D was 2,4-dichlorophenol (2,4-DCP), and degradation of the aromatic nucleus was by intradiol cleavage. Growth lag times prior to the on-set of degradation, and the total time required for degradation, were linearly related to the starting population density and the initial 2,4-D concentration. BRI6001, grown on 2,4-D, oxidized a variety of structurally similar chlorinated aromatic compounds accompanied by stoichiometric chloride release.

2,4-Dichlorophenoxyacetic Acid↗

Stimulation and molecular characterization of bacterial perchlorate degradation by plant-produced electron donors.

Root homogenate from poplar trees (Populus deltoides x nigra DN34, Imperial Carolina) stimulated perchlorate degradation in microcosms of soil and water samples collected at a perchlorate contaminated site, the Longhorn Army Ammunition Plant (LHAAP), located outside Karnack, Texas. Direct use of root products by perchlorate-degrading bacteria was shown for the first time as six pureculture bacteria isolated from LHAAP perchlorate-degrading microcosms degraded perchlorate when given root products as the sole exogenous source of carbon and electron donor. Nonenriched environmental consortia were able to utilize root products for perchlorate degradation, regardless of prior exposure to perchlorate. Microcosms that contained perchlorate-contaminated groundwater (MW-3) or uncontaminated surface water (Harrison Bayou) as inoculum degraded approximately 240 and 160 mg L(-1) perchlorate, respectively, using root products (approximately 440 mg L(-1) as COD) over 38 days. The predominant bacterial species in these aqueous microcosms, identified by DGGE, depended only upon the source inoculum as similar sequences were obtained whether root products or lactate was the electron donor. Sequences from DGGE bands that matched species within Dechloromonas, a genus consisting of many perchlorate degraders, were identified in all perchlorate-degrading microcosms. This study demonstrates the ability of root products to drive perchlorate respiration by bacteria and the potential for successful achievement of perchlorate rhizodegradation using in situ phytoremediation.

Anaerobiosis↗

Degradation of environmental pollutants by Trametes trogii.

The ability of the ligninolytic fungus Trametes trogii to degrade in vitro different xenobiotics (PCBs, PAHs and dyes) was evaluated. Either 200 ppm of a PCB mixture (Aroclor 1150) or 160 ppm of an industrial PAH mixture (10% V/V of PAHs, principal components hexaethylbenzene, naphthalene, 1-methyl naphthalene, acenaphthylene, anthracene, fluorene and phenanthrene), were added to trophophasic and idiophasic cultures growing in a nitrogen limited mineral medium (glucose/asparagine) and in a complex medium (malt extract/glucose). Gas-liquid chromatography proved that within 7 to 12 d more than 90% of the organopollutants added were removed. The decrease in absorbance at 620 nm demonstrated that cultures of this fungus were able to transform 80% of the dye Anthraquinone-blue (added at a concentration of 50 ppm) in 1.5 h. Enzyme estimations indicated high activity of laccase (up to 0.55 U/mL), as well as lower production of manganese-peroxidase. Laccase activity, detected in all the conditions assayed, could be implicated in the degradation of these organopollutants. Considering the results obtained, T. trogii seems promising for detoxification.

Aroclors↗

Bioremediation of creosote-contaminated soil in South Africa by landfarming.

AIMS: To determine the combined effects of biostimulation and bioaugmentation in the landfarming of a mispah form (lithosol; food and Agriculture Organisation (FAO)) soil contaminated with >310000 mg kg-1 creosote with a view to developing a bioremediation technology for soils heavily contaminated with creosote. METHODS AND RESULTS: The excavated soil was mixed with 2500 kg ha-1 dolomitic lime and 2000 kg ha-1 mono-ammonium phosphate (MAP) before spreading over a treatment bed of shale reinforced with clay. Sewage sludge (500 kg) was ploughed into 450 m3 of contaminated soil in the second and sixth months of treatment. A further 1000 kg ha-1 MAP was added to the soil at the end of the fifth month. Moisture was maintained at 70% field capacity. Total creosote was determined by the US Environmental Protection Agency (EPA) method 418.1 and concentrations of selected creosote components were determined by gas chromatography/flame ionisation detection (GC/FID). Total creosote was reduced by more than 90% by the 10th month of landfarming. The rate of reduction in creosote concentration was highest after the addition of sewage sludge. The three-ring PAHs were more slowly removed than naphthalene and the phenolic compounds. The four- and five-ring PAHs, although persist until the end of treatment, were reduced by 76-87% at the end of the experiment. CONCLUSIONS: A combination of biostimulation and bioaugmentation during landfarming could enhance the bioremediation of soils heavily contaminated with creosote. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides information on the management of a combination of biostimulation and bioaugmentation during landfarming, and contributes to the knowledge and database necessary for the development of a technology for bioremediating creosote-contaminated land.

Agriculture↗

Prion degradation in soil: possible role of microbial enzymes stimulated by the decomposition of buried carcasses.

This study is part of a European project focused on understanding the biotic and abiotic mechanisms involved in the retention and dissemination of transmissible spongiform encephalopathies (TSE) infectivity in soil in order to propose practical recommendations to limit environmental contamination. A 1-year field experiment was conducted with lamb carcasses buried in a pasture soil at three depths (25, 45, and 105 cm). Microbial community response to carcasses was monitored through the potential proteolytic activity and substrate induced respiration (SIR). Soil above carcasses and control soil exhibited low proteolytic capacity, whatever the depth of burial. Contrastingly, in soil beneath the carcasses, proteolysis was stimulated. Decomposing carcasses also stimulated SIR, i.e., microbial biomass, suggesting that proteolytic populations specifically developed on lixiviates from animal tissues. Decomposition of soft tissues occurred within 2 months at subsurface while it lasted at least 1 year at deeper depth where proteolytic activities were season-dependent. The ability of soil proteases to degrade the beta form of prion protein was shown in vitro and conditions of burial relevant to minimize the risk of prion protein dissemination are discussed.

Animals↗

Bioremediation: an important alternative for soil and industrial wastes clean-up.

Industrial and environmental biotechnology are going to new paths, resulting in processes with "clean technologies", with the maximum production and the less residues. Technologies of remediation and bioremediation are continuously being improved using genetically modified microorganisms or those naturally occurring, to clean residues and contaminated areas from toxic organics. Bioremediation of soils, water and marine environments has many advantages but at the same time it is a challenge for the researchers and engineers. Consequently, it is extremely important to carry out feasibility study based on pilot-testing before starting a remediation project in order to determine the best conditions for the process. The article presents a brief review of bioremediation including the description of the different methods applied to soil and industrial wastes, and, finally, some experiences of solid-state fermentation in relation to bioremediation.

Bacteria↗