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Cloning, sequencing, and characterization of the hexahydro-1,3,5-Trinitro-1,3,5-triazine degradation gene cluster from Rhodococcus rhodochrous.

Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) is a high explosive which presents an environmental hazard as a major land and groundwater contaminant. Rhodococcus rhodochrous strain 11Y was isolated from explosive contaminated land and is capable of degrading RDX when provided as the sole source of nitrogen for growth. Products of RDX degradation in resting-cell incubations were analyzed and found to include nitrite, formaldehyde, and formate. No ammonium was excreted into the medium, and no dead-end metabolites were observed. The gene responsible for the degradation of RDX in strain 11Y is a constitutively expressed cytochrome P450-like gene, xplA, which is found in a gene cluster with an adrenodoxin reductase homologue, xplB. The cytochrome P450 also has a flavodoxin domain at the N terminus. This study is the first to present a gene which has been identified as being responsible for RDX biodegradation. The mechanism of action of XplA on RDX is thought to involve initial denitration followed by spontaneous ring cleavage and mineralization.

Amino Acid Sequence↗

Incorporating Monte Carlo analysis into multimedia environmental fate models.

We have incorporated a Monte Carlo module into a steady state nonequilibrium level III fugacity model in order to account for variability in degradation half-lives and observe the effects that this variability has on the model outputs. This method leads to a distribution of important output parameters such as the overall persistence time, which can be used in decision-making processes or risk assessment. For herbicides such as atrazine, where the emission rate primarily is to the soil compartment, it is shown that the main contribution to the variability in overall persistence time is the soil degradation half-life.

Atmosphere↗

Prediction of pesticide behavior in soil by means of simple field tests.

A simple method is given for forecasting the reaction under different soil and site conditions of 106 pesticides with known binding strength by humus and clay in relation to pH and with known volatility and aerobic and anaerobic degradability. The method is based on easy-to-determine soil characteristics (texture, humus content, pH, groundwater table) and meteorological data (mean temperature, climatic water balance). It is possible to use this method to predict the environmental risk, especially the risk of contamination, and the groundwater pollution potential of a particular pesticide for a particular site in the field. In addition, recommendations for food contamination are given. This simple approach is suitable for use in on-site consultations. A reduction in pesticide input into near-surface groundwater can be expected when the recommendations derived from the model are applied. The use of this pragmatic model by farmers increases the awareness of the vulnerability of the groundwater recourse.

Adsorption↗

Bioreductive deposition of platinum nanoparticles on the bacterium Shewanella algae.

An environmentally friendly method using the metal ion-reducing bacterium Shewanella algae was proposed to deposit platinum nanoparticles. Resting cells of S. algae were able to reduce aqueous PtCl(6)(2-) ions into elemental platinum at room temperature and neutral pH within 60min when lactate was provided as the electron donor. Biogenic platinum nanoparticles of about 5nm were located in the periplasm--a preferable, cell surface location for easy recovery of biogenic nanoparticles.

Biodegradation, Environmental↗

An easy colorimetric assay for screening and qualitative assessment of deiodination and dehalogenation by bacterial cultures.

Halogenated organic substances are among the main environmental concerns. A number of micro-organisms are able to dehalogenate these compounds. However, the methods for the assessment of micro-organismal ability to dehalogenate are expensive and require complex instrumentation. Here, an easy colorimetric assay for the screening and assessment of the ability of bacterial cultures to deiodinate, and potentially dehalogenate, chemical substances is proposed. The method is based on the oxidation of iodide, released due to biotransformation, to iodine followed by a subsequent detection of iodine by a classical reaction with starch.

Anabaena↗

Removal of Cu, Cr, Ni, Zn, and Cd from electroplating wastes and synthetic solutions by vermicompost of cattle manure.

This study was undertaken to evaluate the retention of Cu, Cr, Ni, Zn, and Cd under laboratory conditions from synthetic solution and electroplating wastes by vermicompost. A glass column was loaded with vermicompost, and metal solutions were passed through it. Metal concentrations were then measured in the eluate in order to evaluate the amounts retained by the vermicompost. Measurements of pH, metal concentrations, moistness, organic matter and ash contents, and infrared and XRD spectroscopy were used for vermicompost characterisation. Vermicompost residues obtained from this process were used for plant nutrition in eroded soil collected from a talus near a highway. Metal retention (in g of metal/kg of vermicompost) from effluents ranged from 2 for Cr and Zn to 4 in the case of Ni. In synthetic solutions, the values for metal retention were 4 for Cd and Zn, 6 for Cu and Ni, and 9 for Cr. The results also showed that metal concentrations in the purified effluents were below the maximum values established for waste discharges into rivers by the Brazilian Environmental Standards. The relatively high available Cd concentration of the vermicompost residue resulted in plant damage. This effect was attributed to the presence of Cd in the synthetic solution passed through the vermicompost. The data obtained do not give a complete picture of using vermicompost in cultivated lands, but such values as are determined do show that it can be suitable to remove heavy metals from industrial effluents.

Animals↗

Dendroremediation of trinitrotoluene (TNT). Part 1: Literature overview and research concept.

BACKGROUND, AIM AND SCOPE: For decades, very large areas of former military sites have been contaminated diffusely with the persistent nitroaromatic explosive 2,4,6-trinitrotoluene (TNT). The recalcitrance of the environmental hazard TNT is to a great extent due to its particulate soil existence, which leads to slow but continuous leaching processes. Although improper handling during the manufacture of TNT seems to be a problem of the past in developed countries, environmental deposition of TNT and other explosives is still going on unfortunately, resulting from thousands of unexploded ordnance or low order explosions at munitions test areas and at current battlefields. OBJECTIVE: Sustainable phytoremediation strategies for explosives in Germany, which intend to use trees to decontaminate soil and groundwater ('dendroremediation'), have to consider that most of the former German military sites are already covered with woodlands, mainly with conifer stands. Therefore, parallel investigation of the remediation potential is necessary for both of the selected hybrids of fast growing broadleaf trees, which are waiting for planting and forest conifers, which have already proven for decades that they are able to grow on explosive contaminated sites. MAIN FEATURES: A short literature review is given regarding phytoremediation of TNT with herbaceous plants and some general aspects of dendroremediation are discussed. Furthermore, an overview of our TNT-dendroremediation research network is introduced, which has the strategic goal to make dendroremediation more calculable for a series of potent trees for site-adapted in situ application and for the assessment of tree remediation potentials in natural attenuation processes. RESULTS AND DISCUSSION: Some of our methods, results and conclusions yet unpublished are presented. For a preliminary calculation of area-related annual TNT dendroremediation potential of five-year-old trees, the following values were assessed: Salix EW-13 6.0, Salix EW-20 8.5, Populus ZP-007 4.2, Betula pendula 5.2, Picea abies 1.9 and Pinus sylvestris 0.8 g m(-2) a(-1). For a 45-year-old spruce forest, an annual natural attenuation potential of 4.2 g TNT m(-2) a(-1) was found. CONCLUSION, RECOMMENDATIONS AND PERSPECTIVE: Our main results deliver quantitative proposals for dendroremediation strategies in situ and provide decision aids. Also aspects of growth of raw materials for energy production are considered. Our dendroremediation research concept for TNT and its congeners can be easily completed for other trees of interest and it can also be applied to herbaceous plants. Knowing the current bottlenecks of phytoremediation and considering the known environmental behaviour of other contaminants, elements of our methodological approach may be easily adapted to those pollutant groups, e.g. for pesticides, pharmaceuticals, PAHs, chlorinated recalcitrants and, with some restrictions, to inorganics and to multiple contaminations. Our dynamical dendrotolerance test systems will help to predict tree growth on polluted areas. To provide some light into the black box of TNT dendroremediation, experimental data regarding the uptake, distribution and degradation of [14C]-TNT in mature tree tissues will be reported in the second part of this publication.

Biodegradation, Environmental↗

Effect of environmental parameters on bacterial degradation of Bunker C oil, Crude oils, and hydrocarbons.

Mixed microbial cultures, previously enriched on Bunker C fuel oil, grew on and degraded Bunker C fuel oil at temperatures ranging from 5 to 28 C. At 15 C, 41 to 85% of the benzene-soluble components of Bunker C disappeared after incubation for 7 days; at 5 C the values ranged from 21 to 52% after 14 days of incubation. A Nocardia sp. isolated from a culture enriched on Bunker C oil grew on Venezuelan crude oil, Bunker C, hexadecane, and a hydrocarbon mixture at temperatures of 5 and 15 C. The 10-C decrease in temperature resulted in an average 2.2-fold decrease in generation time of the bacteria. Gas-liquid chromatographic measurements of Venezuelan and Arabian crude oils which had been incubated with the Nocardia sp. showed significant degradation of the n-alkane portion and the chromatographically unresolved components of the oils. The concentration of elemental nitrogen required to bring about the disappearance of 1 mg of hexadecane by the Nocardia sp. was 0.5 mg. The results confirm suggestions that the rate of natural biodegradation of oil in marine temperate-to-polar zones is probably limited by low temperatures and phosphorus concentrations, but suggest that the concentrations of nitrogen occurring naturally are probably not rate-limiting factors.

Alkanes↗

Fate and occurrence of X-ray contrast media in the environment.

Interest in the presence of pharmaceuticals in the environment has recently increased. Despite continuous research efforts there is still a large gap in our knowledge of their fate and effects on the ecosystem. This review covers current information on the occurrence of iodinated X-ray contrast media (ICM) in the environment and developments in the analysis of these highly polar organic micropollutants in aqueous environmental samples. Findings from monitoring surveys conducted on wastewater-treatment plants (WWTP), surface waters, and drinking waters are compiled, and strategies for removal of the compounds in WWTP and waterworks using advanced treatment are reported. Characteristics and advantages of different compound-specific or element-specific mass spectrometric techniques used to monitor ICM in the environment are compared, and applications in elucidation of the structures of biotransformation products, generated in laboratory-scale experiments that simulate sewage treatment or river water/sediment systems, are described.

Animals↗

Recent developments in molecular techniques for identification and monitoring of xenobiotic-degrading bacteria and their catabolic genes in bioremediation.

The pollution of soil and water with xenobiotics is widespread in the environment and is creating major health problems. The utilization of microorganisms to clean up xenobiotics from a polluted environment represents a potential solution to such environmental problems. Recent developments in molecular-biology-based techniques have led to rapid and accurate strategies for monitoring, discovery and identification of novel bacteria and their catabolic genes involved in the degradation of xenobiotics. Application of these techniques to bioremediation has also improved our understanding of the composition, phylogeny, and physiology of metabolically active members of the microbial community in the environment. This review provides an overview of recent developments in molecular-biology-based techniques and their application in bioremediation of xenobiotics.

Bacteria↗

Arsenic resistance and removal by marine and non-marine bacteria.

Arsenic resistance and removal was evaluated in nine bacterial strains of marine and non-marine origins. Of the strains tested, Marinomonas communis exhibited the second-highest arsenic resistance with median effective concentration (EC(50)) value of 510 mg As l(-1), and was capable of removing arsenic from culture medium amended with arsenate. Arsenic accumulation in cells amounted to 2290 microg As g(-1) (dry weight) when incubated on medium containing 5 mg As l(-1) of arsenate. More than half of the arsenic removed was related to metabolic activity: 45% of the arsenic was incorporated into the cytosol fraction and 10% was found in the lipid-bound fraction of the membrane, with the remaining arsenic considered to be adsorbed onto the cell surface. Potential arsenic resistance and removal were also examined in six marine and non-marine environmental water samples. Of the total bacterial colony counts, 28-100% of bacteria showed arsenic resistance. Some of the bacterial consortia, especially those from seawater enriched with arsenate, exhibited higher accumulated levels of arsenic than M. communis under the same condition. These results showed that arsenic resistant and/or accumulating bacteria are widespread in the aquatic environment, and that arsenic-accumulating bacteria such as M. communis are potential candidates for bioremediation of arsenic contaminated water.

Arsenates↗

Genetically engineered phytoremediation: one man's trash is another man's transgene.

Plant-based environmental remediation, or phytoremediation, has been widely pursued in recent years as a favorable clean-up technology and is an area of intensive scientific investigation. For the vast majority of field applications, vegetative 'phyto-crops' are selected specifically for their capacity for site decontamination and not for additional concurrent or post-remediation utility. By contrast, a recent publication by Ellis and colleagues highlights potential anti-carcinogenic uses for plants genetically engineered primarily for detoxification of selenium-polluted soils and sediments.

Anticarcinogenic Agents↗

Operational characteristics of effective removal of H2S and NH3 waste gases by activated carbon biofilter.

Simultaneous removal of hydrogen sulfide (H2S) and ammonia (NH3) gases from gaseous streams was studied in a biofilter packed with granule activated carbon. Extensive studies, including the effects of carbon (C) source on the growth of inoculated microorganisms and gas removal efficiency, product analysis, bioaerosol emission, pressure drop, and cost evaluation, were conducted. The results indicated that molasses was a potential C source for inoculated cell growth that resulted in removal efficiencies of 99.5% for H2S and 99.2% for NH3. Microbial community observation by scanning electron microscopy indicated that granule activated carbon was an excellent support for microorganism attachment for long-term waste gas treatment. No disintegration or breakdown of biofilm was found when the system was operated for 140 days. The low bioaerosol concentration emitted from the biofilter showed that the system effectively avoided the environmental risk of bioaerosol emission. Also, the system is suitable to apply in the field because of its low pressure drop and treatment cost. Because NH3 gas was mainly converted to organic nitrogen, and H2S gas was converted to elemental sulfur, no acidification or alkalinity phenomena were found because of the metabolite products. Thus, the results of this study demonstrate that the biofilter is a feasible bioreactor in the removal of waste gases.

Adsorption↗

Phytoremediation potential of Spirulina (Arthrospira) platensis: biosorption and toxicity studies of cadmium.

This study examines the possibility of using Spirulina (Arthrospira) platensis TISTR 8217 to remove low concentrations of cadmium (less than 100 mg/l) from wastewater. The cyanobacteria were exposed to six different cadmium concentrations for 96 h, and the growth rate was determined using an optical density at 560 nm. The inhibiting concentration (IC50) was estimated using probit analysis. The IC50 at 24, 48, 72, and 96 h were 13.15, 16.68, 17.28, and 18.35 mg/l Cd, respectively. Cellular damage was studied under a light microscope and a transmission electron microscope. Swollen cells and fragmented filaments were observed. Cell injury increased with increasing concentrations of cadmium. Ultrastructural changes were observed in the algae exposed to cadmium concentrations both close to IC50 (14.68 mg/l) and at IC50 (18.35 mg/l). The alterations induced by cadmium were disintegration and disorganization of thylakoid membranes, presence of large intrathylakoidal space, increase of polyphosphate bodies, and cell lysis. In addition, the cadmium adsorption by algal cells was studied. Environmental factors were found to have an effect on biosorption. The uptake of cadmium was not affected by the temperature of the solution, but the sorption was pH dependent. The optimum pH for biosorption of algal cells was 7. The cadmium uptake process was rapid, with 78% of metal sorption completed within 5 min. The sorption data fit well to the Langmuir isotherm. The maximum adsorption capacity for S. platensis was 98.04 mg Cd per g biomass.

Adsorption↗

Sorption of phenanthrene to environmental black carbon in sediment with and without organic matter and native sorbates.

Strong sorption to soot- and charcoal-like material (collectively termed black carbon or BC) in soils and sediments is possibly the reason for recent observations of elevated geosorbent-water distribution ratios, slow desorption, limited uptake, and restricted bioremediation. We evaluated the role of environmental BC in the sorption of phenanthrene (PHE) to a polluted lake sediment from a Rhine River sedimentation area. Sorption isotherms were determined over a wide concentration range (0.0005-6 microg/ L) for the original sediment (with organic matter or OM, native sorbates, and BC), sediment from which we had stripped > 90% of the native sorbates (only OM and BC), and sediment combusted at 375 degrees C (only BC). The sorption isotherms of the original and stripped sediments were almost linear (Freundlich coefficient or n(F) > 0.9), whereas the isotherm of the BC remaining after the sediment combustion was highly nonlinear (n(F) = 0.54). At low concentrations (ng/L range), PHE sorption to BC in the combusted sediment was found to exceed the total PHE sorption in the original and stripped sediments. This implies that it may not be possible to use a BC-water sorption coefficient measured in combusted sediment to estimate total sorption to the original sediment. This "intrinsic" BC-water sorption coefficient after combustion was calculated to be 9 times larger than the "environmental" one in the untreated sediment. Competition between the added PHE and the native PAHs and/or OM may explain this difference. It appears that, at low aqueous PHE concentrations (ng/L and below), BC is the most important geosorbent constituent with respect to sorption. At higher concentrations (microg/L), BC sorption sites become saturated and BC sorption is overwhelmed by sorption to the other OM constituents. Because sorption is a central process affecting contaminant behavior and ecotoxicity, understanding this process can strongly contribute to risk assessment and fate modeling.

Adsorption↗

Needs and strategies for genetic control: municipal wastes.

Municipal wastewater, sludge, and refuse constitute major material flows and area of public concern. Substantial lists of desired improvements in the biological treatment of each waste can be formulated. Many improvements can be brought about by better application of biological and process fundamentals; widespread application of good process control and existing microorganisms is still not a reality. Nevertheless, several improvements seem amenable to genetic control techniques. Whether the genetic control technique involves selection of a novel microorganism or genetic manipulation of conventional or novel microorganisms, two key requirements stand out. First, we must recognize the physiological factor that provides the desired function. Second, we must recombine the genes for the desired function into a microorganism that can thrive in a treatment process. For environmental applications, identifying the physiological effect and an appropriate microorganism are the most critical steps. However, they are probably the least well developed, as biochemists and genetic engineers have concentrated mainly on isolating, recombining, and expressing relatively well known genes into convenient laboratory strains. Because of the many different steps associated with applications of genetic control to municipal-waste treatment, the successful application requires coordination of biochemistry, microbiology, and process engineering. No one discipline has all the requisite tools and insight to do the whole job.

Bacterial Physiological Phenomena↗

Isolation of a member of Acinetobacter species involved in atrazine degradation.

Contribution of Acinetobacter genus in the degradation of atrazine and its analogs is reported here. An interesting bacterial isolate capable of degrading atrazine as high as 250 ppm was isolated from a soil heavily contaminated with atrazine. The permissible level of atrazine in drinking water is 3 ppb and hence use of a strain capable of atrazine degradation as high as 250 ppm would be of immense help for rapid environmental cleanup. This isolate was found to be capable of best growth at 37 degrees C and at pH inclined towards the alkaline side. It was found that atrazine was utilized as a carbon and not as a nitrogen source. Acinetobacter species was also active on other triazine pesticides, viz., simazine, terbutryn, cyanazine, and prometon. There are very few reports on the degradation of atrazine by any member of this genus and hence this could lead to new degradation pathways and new metabolites.

Acinetobacter↗

DsrB gene-based DGGE for community and diversity surveys of sulfate-reducing bacteria.

A denaturing gradient gel electrophoresis (DGGE) method was developed to assess the diversity of dsrB (dissimilatory sulfite reductase beta-subunit)-genes in sulfate-reducing communities. For this purpose a PCR primer pair was optimized for the amplification of a approximately 350 bp dsrB gene fragment that after DGGE gel electrophoresis enabled us to discriminate between dsrB genes of different SRB-subgroups,-genera and -species. The dsrB-DGGE method revealed considerable genetic diversity when applied to DNA extracts obtained from aquifer samples that were derived from monitoring wells of an in situ metal precipitation (ISMP) pilot project conducted at the site of a non-ferrous industry or from environmental heavy metal contaminated samples. The sequences of the excised and sequenced DGGE bands represented dsrB genes of different SRB-subgroups,-genera and -species, thus confirming the broad applicability of the PCR primer pair. Linking the results of the physico-chemical follow-up of the field and lab experiments to the dsrB-DGGE data will provide a better understanding of the contribution of the SRB populations to the ongoing ISMP processes.

Base Sequence↗