Unravelling the process of petroleum hydrocarbon biodegradation in different filter materials of constructed wetlands by stable isotope fractionation and labelling studies.

Maintaining and supporting complete biodegradation during remediation of petroleum hydrocarbon contaminated groundwater in constructed wetlands is vital for the final destruction and removal of contaminants. We aimed to compare and gain insight into biodegradation and explore possible limitations in different filter materials (sand, sand amended with biochar, expanded clay). These filters were collected from constructed wetlands after two years of operation and batch experiments were conducted using two stable isotope techniques; (i) carbon isotope labelling of hexadecane and (ii) hydrogen isotope fractionation of decane. Both hydrocarbon compounds hexadecane and decane were biodegraded. The mineralization rate of hexadecane was higher in the sandy filter material (3.6 µg CO2 g-1 day-1) than in the expanded clay (1.0 µg CO2 g-1 day-1). The microbial community of the constructed wetland microcosms was dominated by Gram negative bacteria and fungi and was specific for the different filter materials while hexadecane was primarily anabolized by bacteria. Adsorption / desorption of petroleum hydrocarbons in expanded clay was observed, which might not hinder but delay biodegradation. Very few cases of hydrogen isotope fractionation were recorded in expanded clay and sand & biochar filters during decane biodegradation. In sand filters, decane was biodegraded more slowly and hydrogen isotope fractionation was visible. Still, the range of observed apparent kinetic hydrogen isotope effects (AKIEH = 1.072-1.500) and apparent decane biodegradation rates (k = - 0.017 to - 0.067 day-1) of the sand filter were low. To conclude, low biodegradation rates, small hydrogen isotope fractionation, zero order mineralization kinetics and lack of microbial biomass growth indicated that mass transfer controlled biodegradation.

Determination of carbon isotope enrichment factors of cis-dichloroethene after precursor amendment.

RATIONALE: Bacterial reductive dechlorination of the groundwater contaminant tetrachloroethene (PCE) involves the formation of lower chlorinated metabolites. Metabolites can be instantaneously formed and consumed in this sequential process; quantification and validation of their isotopic effects conventionally rely on separate laboratory microcosm studies. Here, we present an evaluation method enabling the determination of the carbon isotope enrichment factor (ε) for the intermediate cis-dichloroethene (cis-DCE) by a single laboratory microcosm study initially amending the precursor PCE only. METHODS: Environmental samples harboring organohalide-respiring bacteria were incubated under anaerobic conditions and then successively and repeatedly amended with PCE and cis-DCE in two separate laboratory microcosm studies. Reductive dechlorination was monitored by analyzing liquid samples using Purge-and-Trap gas chromatography isotope ratio mass spectrometry GC/MS-C/IRMS. The prerequisites of the presented evaluation method are mass and δ-value balancing. The evaluation method was validated by agglomerative hierarchical classification of Rayleigh plot data points. RESULTS: The sample-sensitive range of εcis-DCE extended from -10.6 ± 0.2‰ to -26.8 ± 0.6‰ (R2 ≥98%). The maximum standard deviations of εcis-DCE were ±1.8‰ for single microcosms, ±1.8‰ for replicates and ±1.0‰ for the compiled replicate data of PCE and cis-DCE amendments. A linear regression of the εcis-DCE for replicates obtained by each amendment study showed a slope of 95% (5 of the 7 data points are within a 95% confidence interval), demonstrating factor congruency and the practicability of the evaluation method. CONCLUSIONS: We found metabolite degradation and formation to be sequential but also stepwise during bacterial reductive dechlorination. The stepwise phases of the degradation of the intermediate eliminate the impact of instantaneous precursor degradation. These stepwise sections were used to determine εcis-DCE -values. Our results showed the validity of εcis-DCE -values over a wide range at initial precursor degradation (PCE). The presented evaluation method could substantially decrease lab costs for microcosm studies designed for εcis-DCE determinations. Moreover, the results indicated that the evaluation method can be applied to other PCE-metabolites.

Transpiration and metabolisation of TCE by willow plants - a pot experiment.

Willows were grown in glass cylinders filled with compost above water-saturated quartz sand, to trace the fate of TCE in water and plant biomass. The experiment was repeated once with the same plants in two consecutive years. TCE was added in nominal concentrations of 0, 144, 288, and 721 mg l(-1). Unplanted cylinders were set-up and spiked with nominal concentrations of 721 mg l(-1) TCE in the second year. Additionally, (13)C-enriched TCE solution (δ(13)C = 110.3 ‰) was used. Periodically, TCE content and metabolites were analyzed in water and plant biomass. The presence of TCE-degrading microorganisms was monitored via the measurement of the isotopic ratio of carbon ((13)C/(12)C) in TCE, and the abundance of (13)C-labeled microbial PLFAs (phospholipid fatty acids). More than 98% of TCE was lost via evapotranspiration from the planted pots within one month after adding TCE. Transpiration accounted to 94 to 78% of the total evapotranspiration loss. Almost 1% of TCE was metabolized in the shoots, whereby trichloroacetic acid (TCAA) and dichloroacetic acid (DCAA) were dominant metabolites; less trichloroethanol (TCOH) and TCE accumulated in plant tissues. Microbial degradation was ruled out by δ(13)C measurements of water and PLFAs. TCE had no detected influence on plant stress status as determined by chlorophyll-fluorescence and gas exchange.

The relative abundance and seasonal distribution correspond with the sources of polycyclic aromatic hydrocarbons (PAHs) in the surface sediments of Chenab River, Pakistan.

Chenab River is one of the most important rivers of Punjab Province (Pakistan) that receives huge input of industrial effluents and municipal sewage from major cities in the Central Punjab, Pakistan. The current study was designed to evaluate the concentration levels and associated ecological risks of USEPA priority polycyclic aromatic hydrocarbons (PAHs) in the surface sediments of Chenab River. Sampling was performed from eight (n = 24) sampling stations of Chenab River and its tributaries. We observed a relatively high abundance of ∑16PAHs during the summer season (i.e. 554 ng g(-1)) versus that in the winter season (i.e. 361 ng g(-1)), with an overall abundance of two-, five- and six-ring PAH congeners. Results also revealed that the nitrate and phosphate contents in the sediments were closely associated with low molecular weight (LMW) and high molecular weight (HMW) PAHs, respectively. Source apportionment results showed that the combustion of fossil fuels appears to be the key source of PAHs in the study area. The risk quotient (RQ) values indicated that seven PAH congeners (i.e. phenanthrene, anthracene, fluoranthene, pyrene, benzo(a)pyrene, chrysene and benzo(a)anthracene) could pose serious threats to the aquatic life of the riverine ecosystem in Pakistan.

Diverse bacteria colonizing leaves and the rhizosphere of lettuce degrade azoxystrobin.

Concerns about the possible effects of pesticide residues on both the environment and human health have increased worldwide. Bioremediation by the use of microorganisms to degrade or remove these residues has emerged as a powerful technology. However, the knowledge about the potential of different microorganisms for pesticide degradation is limited. This study focused on the isolation and characterisation of bacterial strains with the potential to degrade the active fungicide ingredient azoxystrobin. Potential degrading bacteria were tested in vitro and in the greenhouse, and the genomes of the best degrading strains were sequenced and analysed. We identified and characterised 59 unique bacterial strains, which were further tested in vitro and in greenhouse trials for their degradation activity. The best degraders from a foliar application trial in the greenhouse were identified as Bacillus subtilis strain MK101, Pseudomonas kermanshahensis strain MK113 and Rhodococcus fascians strain MK144 and analysed by whole genome sequencing. Genome analysis revealed that these three bacterial strains encode several genes predicted to be involved in the degradation of pesticides e.g., benC, pcaG, pcaH, however we could not find any specific gene previously reported to be involved in azoxystrobin degradation e.g., strH. Genome analysis pinpointed to some potential activities involved in plant growth promotion.

Transcriptome analysis of nitrate assimilation in Aspergillus nidulans reveals connections to nitric oxide metabolism.

Nitrate is a dominant form of inorganic nitrogen (N) in soils and can be efficiently assimilated by bacteria, fungi and plants. We studied here the transcriptome of the short-term nitrate response using assimilating and non-assimilating strains of the model ascomycete Aspergillus nidulans. Among the 72 genes positively responding to nitrate, only 18 genes carry binding sites for the pathway-specific activator NirA. Forty-five genes were repressed by nitrate metabolism. Because nirA(-) strains are N-starved at nitrate induction conditions, we also compared the nitrate transcriptome with N-deprived conditions and found a partial overlap of differentially regulated genes between these conditions. Nitric oxide (NO)-metabolizing flavohaemoglobins were found to be co-regulated with nitrate assimilatory genes. Subsequent molecular characterization revealed that the strongly inducible FhbA is required for full activity of nitrate and nitrite reductase enzymes. The co-regulation of NO-detoxifying and nitrate/nitrite assimilating systems may represent a conserved mechanism, which serves to neutralize nitrosative stress imposed by an external NO source in saprophytic and pathogenic fungi. Our analysis using membrane-permeable NO donors suggests that signalling for NirA activation only indirectly depends on the nitrate transporters NrtA (CrnA) and NrtB (CrnB).

Inhibition and stimulation of two perchloroethene degrading bacterial cultures by nano- and micro-scaled zero-valent iron particles.

The pollutant perchloroethene (PCE) can often be found at urban contaminated sites. Thus in-situ clean-up methods, like remediation using zero valent iron (ZVI) or bacterial dechlorination, are preferred. During the remediation with ZVI particles anaerobic corrosion occurs as an unwanted, particle consuming side reaction with water. However, in this reaction H2 is formed, which is usually scarce during anaerobic microbial dechlorination. Dehalococcoides needs H2 for cell growth using it as an electron donor to dechlorinate chlorinated hydrocarbons. Combining application of ZVI with bacterial dechlorination can turn ZVI in a H2 donor leading to a more controllable bacterial dechlorination, a smaller amount of ZVI suspension and decreased remediation costs. In this study nano- and micro scaled ZVI particles (nZVI, mZVI) were combined in microcosms with two dechlorinating bacterial cultures. The two cultures showed different dechlorination behaviors with ethene and cis-DCE as final products. Phospholipid fatty acids (PLFA) associated with Dehalococcoides (18:1w7, 18:1w7c, 10:Me16:0) and Geobacteriaceae (16,1w7c; 15:0; 16:0) have been found in both bacterial cultures, slight differences in their abundance could explain the different dechlorinating behaviors. The combination of both bacterial cultures with mZVI led to a stimulated dechlorination process leading to about two times higher kobs for PCE dechlorination (0.01-0.05 h-1). In the otherwise cis-DCE accumulating culture complete dechlorination to ethene was achieved. While addition of nZVI inhibited both cultures. Combined with nZVI the completely dechlorinating culture produced lower amounts of dechlorinated products (3.2 µmol) as compared to the single biotic treatment (5.1 µmol). Combining the incompletely dechlorinating culture with nZVI significantly reduced the kobs,PCE (single: 8 × 10-3 ± 3 × 10-4 h-1; combination: 5 × 10-3 ± 2 × 10-4 h-1). H2 produced by nZVI and mZVI was utilized by both bacterial cultures. The particle size, resulting specific surface areas, agglomeration tendencies and reactivity appears to be crucial for the effect on microbial cells.

Synergistic effects of microbial anaerobic dechlorination of perchloroethene and nano zero-valent iron (nZVI) - A lysimeter experiment.

Perchloroethene (PCE) is a hazardous and persistent groundwater pollutant. Both treatment with nanoscaled zero-valent iron (nZVI) and biological degradation by bacteria have downsides. Distribution of nZVI underground is difficult and a high percentage of injected nZVI is consumed by anaerobic corrosion, forming H2 rather than being available for PCE dechlorination. On the other hand, microbial PCE degradation can suffer from the absence of H2. This can cause the accumulation of the hazardous metabolites cis-1,2-dichloroethene (DCE) or vinylchloride (VC). The combination of chemical and biological PCE degradation is a promising approach to overcome the disadvantages of each method alone. In this lysimeter study, artificial aquifers were created to test the influence of nZVI on anaerobic microbial PCE dechlorination by a commercially available culture containing Dehalococcoides spp. under field-like conditions. The effect of the combined treatment was investigated with molasses as an additional electron source and after cessation of molasses addition. The combination of nZVI and the Dehalococcoides spp. containing culture led to a PCE discharge in the lysimeter outflow that was 4.7 times smaller than that with nZVI and 1.6 times smaller than with bacterial treatment. Moreover, fully dechlorinated end-products showed an 11-fold increase compared to nZVI and a 4.2-fold increase compared to the microbial culture. The addition of nZVI to the microbial culture also decreased the accumulation of hazardous metabolites by 1.7 (cis-DCE) and 1.2 fold (VC). The stimulatory effect of nZVI on microbial degradation was most obvious after the addition of molasses was stopped.

Core-Shell Fe/FeS Nanoparticles with Controlled Shell Thickness for Enhanced Trichloroethylene Removal.

Zero-valent iron nanoparticles (nZVI) treated by reduced sulfur compounds (i.e., sulfidated nZVI, S-nZVI) have attracted increased attention as promising materials for environmental remediation. While the preparation of S-nZVI and its reactions with various groundwater contaminants such as trichloroethylene (TCE) were already a subject of several studies, nanoparticle synthesis procedures investigated so far were suited mainly for laboratory-scale preparation with only a limited possibility of easy and cost-effective large-scale production and FeS shell property control. This study presents a novel approach for synthesizing S-nZVI using commercially available nZVI particles that are treated with sodium sulfide in a concentrated slurry. This leads to S-nZVI particles that do not contain hazardous boron residues and can be easily prepared off-site. The resulting S-nZVI exhibits a core-shell structure where zero-valent iron is the dominant phase in the core, while the shell contains mostly amorphous iron sulfides. The average FeS shell thickness can be controlled by the applied sulfide concentration. Up to a 12-fold increase in the TCE removal and a 7-fold increase in the electron efficiency were observed upon amending nZVI with sulfide. Although the FeS shell thickness correlated with surface-area-normalized TCE removal rates, sulfidation negatively impacted the particle surface area, resulting in an optimal FeS shell thickness of approximately 7.3 nm. This corresponded to a particle S/Fe mass ratio of 0.0195. At all sulfide doses, the TCE degradation products were only fully dechlorinated hydrocarbons. Moreover, a nearly 100% chlorine balance was found at the end of the experiments, further confirming complete TCE degradation and the absence of chlorinated transformation products. The newly synthesized S-nZVI particles thus represent a promising remedial agent applicable at sites contaminated with TCE.

Soil amendments and cultivar selection can improve rice yield in salt-influenced (tsunami-affected) paddy fields in Sri Lanka.

The tsunami disaster in the Indian Ocean in December 2004 caused devastation of agricultural soils by salt water over wide areas. Many rice fields located close to the coast were affected by the flood of seawater. Electric conductivity (EC) of soils in tsunami-affected rice fields was found to be higher compared to unaffected fields 2 years after the tsunami. Four soil amendments (gypsum, dolomite, cinnamon ash and rice-husk-charcoal) were tested for their influence on improving the yield parameters of rice grown in a tsunami-affected and a non-affected area. Yield parameters were compared with an untreated control of the same cultivar (AT362) and with a salt resistant rice variety (AT354). The salt resistant variety had the highest grain yield. The two amendments gypsum and rice-husk-charcoal led to an increase in grain yield compared to the untreated control, whereas dolomite and cinnamon ash had no significant effect on grain yield.

Differentiation between physical and chemical effects of oil presence in freshly spiked soil during rhizoremediation trial.

Petroleum contamination and its remediation via plant-based solutions have got increasing attention by environmental scientists and engineers. In the current study, the physiological and growth responses of two diesel-tolerant plant species (tolerance limit: 1500-2000 mg/kg), Italian ryegrass (Lolium multiflorum) and Birdsfoot trefoil (Lotus corniculatus), have been investigated in vegetable oil- and diesel oil-amended soils. A long-term (147-day) greenhouse pot experiment was conducted to differentiate the main focus of the study: physical and chemical effects of oil (vegetable and diesel) in freshly spiked soils via evaluating the plant performance and hydrocarbon degradation. Moreover, plant performance was evaluated in terms of seed germination, plant shoot biomass, physiological parameters, and root biomass. Addition of both diesel oil and vegetable oil in freshly spiked soils showed deleterious effects on seedling emergence, root/shoot biomass, and chlorophyll content of grass and legume plants. Italian ryegrass showed more sensitivity in terms of germination rate to both vegetable and diesel oil as compared to non-contaminated soils while Birdsfoot trefoil reduced the germination rate only in diesel oil-impacted soils. The results of the current study suggest that both physical and chemical effects of oil pose negative effects of plant growth and root development. This observation may explain the phenomenon of reduced plant growth in aged/weathered contaminated soils during rhizoremediation experiments.

Impact of sorption processes on PCE concentrations in organohalide-respiring aquifer sediment samples.

The evaluation of groundwater contaminant e.g. tetrachloroethene (PCE) degradation processes requires complete quantification of and pathway analysis of the groundwater contaminant under investigation. For example the reduction of PCE concentrations in the groundwater by unknown dissolution and/or sorption processes will impede interpretation of the fate and behaviour of such contaminants. In the present study PCE dissolution and sorption processes during anaerobic microbial degradation of chlorinated ethenes were investigated. For this purpose, microcosms were prepared using sediment samples from a PCE-contaminated aquifer, which in previous studies had demonstrated anaerobic organohalide respiration of PCE. Solid/water distribution coefficients (kd) of PCE were determined and validated by loss-on-ignition (LOI) and PCE sorption experiments. The determined kd magnitudes indicated methodological congruency, yielding values for sediment samples within a range of 1.15±0.02 to 5.93±0.34L·kg-1. The microcosm experiment showed lower PCE concentrations than expected, based on spiked PCE and observed anaerobic microbial degradation processes. Nevertheless the amount of PCE spike added was completely recovered albeit in the form of lower chlorinated metabolites. A delay due to dissolution processes was not responsible for this phenomenon. Sorption to sediments could only partially explain the reduction of PCE in the water phase. Accordingly, the results point to reversible sorption processes of PCE, possibly onto bacterial cell compartments and/or exopolymeric substances.

Degradation of polycyclic aromatic hydrocarbons in a mixed contaminated soil supported by phytostabilisation, organic and inorganic soil additives.

In soil, mixed contamination with potentially toxic trace elements and polycyclic aromatic hydrocarbons (PAHs) may persist for a long time due to strong adsorption to the soil matrix and to its toxicity to microorganism. We conducted an incubation batch experiment to test the effect of soil amendments (biochar, gravel sludge, iron oxides) on the immobilisation of trace elements. To monitor microbial degradation, a 13C-PHE (phenanthrene) label was introduced to soil for 13C-PLFA (phospholipid fatty acid) analysis. Soil amendments increased soil pH, reduced mobility of NH4NO3-extractable trace elements Cd and Zn, and increased mobile Cu. A small consortium of PHE degraders was identified mainly in the microbial groups of gram-negative bacteria and actinomycetes. The degradation process of PHE peaked 9days after incubation start. PAH concentrations remained constant in the soil within the 30-day incubation, except for the easily available 13C-PHE in the amended treatment. In order to test the effect of plants and soil amendments under more realistic conditions, we also conducted an outdoor pot experiment with black locust (Robinia pseudoacacia Nyirsegi). Furthermore, soil amendments increased the mobility of soil Cu and As and decreased the mobility of Cd, Pb and Sb. The uptake of trace elements to leaves was low. Σ 16 U.S. EPA PAHs were significantly reduced only in the combined treatment of black locust and soil amendments after 12months of plant growth. Soil amendment-assisted phytoremediation showed a high efficiency in PAH dissipation and may be a useful remediation technique for mixed contaminated soils.

Investigations of microbial degradation of polycyclic aromatic hydrocarbons based on 13C-labeled phenanthrene in a soil co-contaminated with trace elements using a plant assisted approach.

Co-contaminations of soils with organic and inorganic pollutants are a frequent environmental problem. Due to their toxicity and recalcitrance, the heterogeneous pollutants may persist in soil. The hypothesis of this study was that degradation of polycyclic aromatic hydrocarbons (PAHs) is enhanced if heavy metals in soil are immobilized and their bioavailability reduced. For metal immobilization and enhanced biodegradation, distinct mineral and organic soil amendments (iron oxides, gravel sludge, biochar) were deployed in an incubation batch experiment. The second part of the experiment consisted of a greenhouse pot experiment applying fast-growing and pollution-tolerant woody plants (willow and black locust). Soil amendments initially immobilized NH4NO3-extractable zinc, cadmium, and lead; after 100 days of incubation, soil amendments showed reductions only for cadmium and a tendency to enhance arsenic mobility. In order to monitor the remediation success, a 13C-phenanthrene (PHE) label was applied. 13C-phospholipid fatty acid analysis (13C-PLFA) further enabled the identification of PHE-degrading soil microorganisms. Both experiments exhibited a similar PLFA profile. Gram-negative bacteria (esp. cy17:0, 16:1ω7 + 6, 18:1ω7c) were the most significant microbial group taking up 13C-PHE. Plants effectively increased the label uptake by gram-positive bacteria and increased the biomass of the fungal biomarker, although their contribution to the degradation process was minor. Plants tended to prolong PAH dissipation in soil; at the end of the experiment, however, all treatments showed equally low total PAH concentrations in soil. While black locust plants tended not to take up potentially toxic trace elements, willows accumulated them in their leaves. The results of this study show that the chosen treatments did not enhance the remediation of the experimental soil.

Microbe and plant assisted-remediation of organic xenobiotics and its enhancement by genetically modified organisms and recombinant technology: A review.

Environmental problems such as the deterioration of groundwater quality, soil degradation and various threats to human, animal and ecosystem health are closely related to the presence of high concentrations of organic xenobiotics in the environment. Employing appropriate technologies to remediate contaminated soils is crucial due to the site-specificity of most remediation methods. The limitations of conventional remediation technologies include poor environmental compatibility, high cost of implementation and poor public acceptability. This raises the call to employ biological methods for remediation. Bioremediation and microbe-assisted bioremediation (phytoremediation) offer many ecological and cost-associated benefits. The overall efficiency and performance of bio- and phytoremediation approaches can be enhanced by genetically modified microbes and plants. Moreover, phytoremediation can also be stimulated by suitable plant-microbe partnerships, i.e. plant-endophytic or plant-rhizospheric associations. Synergistic interactions between recombinant bacteria and genetically modified plants can further enhance the restoration of environments impacted by organic pollutants. Nevertheless, releasing genetically modified microbes and plants into the environment does pose potential risks. These can be minimized by adopting environmental biotechnological techniques and guidelines provided by environmental protection agencies and other regulatory frameworks. The current contribution provides a comprehensive overview on enhanced bioremediation and phytoremediation approaches using transgenic plants and microbes. It also sheds light on the mitigation of associated environmental risks.

Isotopic effects of PCE induced by organohalide-respiring bacteria.

Reductive dechlorination performed by organohalide-respiring bacteria (OHRB) enables the complete detoxification of certain emerging groundwater pollutants such as perchloroethene (PCE). Environmental samples from a contaminated site incubated in a lab-scale microcosm (MC) study enable documentation of such reductive dechlorination processes. As compound-specific isotope analysis is used to monitor PCE degradation processes, nucleic acid analysis-like 16S-rDNA analysis-can be used to determine the key OHRB that are present. This study applied both methods to laboratory MCs prepared from environmental samples to investigate OHRB-specific isotope enrichment at PCE dechlorination. This method linkage can enhance the understanding of isotope enrichment patterns of distinct OHRB, which further contribute to more accurate evaluation, characterisation and prospection of natural attenuation processes. Results identified three known OHRB genera (Dehalogenimonas, Desulfuromonas, Geobacter) in diverse abundance within MCs. One species of Dehalogenimonas was potentially involved in complete reductive dechlorination of PCE to ethene. Furthermore, the isotopic effects of PCE degradation were clustered and two isotope enrichment factors (ε) (- 11.6‰, - 1.7‰) were obtained. Notably, ε values were independent of degradation rates and kinetics, but did reflect the genera of the dechlorinating OHRB.

Are the biological properties of kaempferol determined by its oxidation products?

Although flavonoid molecules have attracted considerable interest in recent years because of their antioxidant effect, there are considerable differences in their chemical properties. Electron paramagnetic resonance (EPR) spectroscopy was used to compare the oxidative free radical chemistry of two such molecules, kaempferol and luteolin, which have the same empirical formula but differ in the position of one OH group. Whereas the basic flavonoid structure remains intact in luteolin, structural changes occur in kaempferol after one-electron oxidation. Autoxidation of kaempferol in alkaline solution and oxidation by at pH 7 led to rapid fragmentation. In contrast, oxidation by horseradish peroxidase/hydrogen peroxide, xanthine/xanthine oxidase (X/XO) or a Fenton reaction system produced a radical whose structure appeared to be based on dimerisation of either the original or a fragment of the flavonoid. Hence, the biological properties of kaempferol are likely to be determined by the chemistry of its oxidation products.

Free radical generation in Pinus sylvestris and Larix decidua seeds primed with polyethylene glycol or potassium salt solutions.

Electron paramagnetic resonance (EPR) spectra of Pinus sylvestris and Larix decidua seeds show that priming with PEG+200 mg kg(-1) gibberelic acid (GA(3)) results in appreciably higher free radical contents than in unprimed control samples. Only relatively minor changes in the free radical levels were observed in seeds primed with K(+) salts. However, both priming treatments have been reported previously to result in faster germination rates compared to controls without changing the germination percentage. In measurements on individual seeds of L. decidua, there were no significant differences between the mean free radical levels in viable and non-viable seeds within each treatment group. Thus, the elevation in free radical levels in the PEG+GA(3) treatments appear to be a direct consequence of the priming treatment and do not correspond to the initiation of germination.

Electron efficiency of nZVI does not change with variation of environmental parameters.

Nanoscale zero-valent iron particles (nZVI) are already applied for in-situ dechlorination of halogenated organic contaminants in the field. We performed batch experiments whereby trichloroethene (TCE) was dehalogenated by nZVI under different environmental conditions that are relevant in practice. The tested conditions include different ionic strengths, addition of polyelectrolytes (carboxymethylcellulose and ligninsulphonate), lowered temperature, dissolved oxygen and different particle contents. Particle properties were determined by Mössbauer spectroscopy, XRD, TEM, SEM, AAS and laser obscuration time measurements. TCE dehalogenation and H2 evolution were decelerated by reduced ionic strength, addition of polyelectrolytes, temperature reduction, the presence of dissolved oxygen and reduced particle content. The partitioning of released electrons between reactions with the contaminant vs. with water (selectivity) was low, independent of the tested conditions. Basically out of hundred electrons that were released via nZVI oxidation only 3.1±1.4 were used for TCE dehalogenation. Even lower selectivities were observed at TCE concentrations below 3.5 mg l(-1), hence particle modifications and/or combination of nZVI with other remediation technologies seem to be necessary to reach target concentrations for remediation. Our results suggest that selectivity is particle intrinsic and not as much condition dependent, hence particle synthesis and potential particle modifications of nZVI particles may be more important for optimization of the pollutant degradation rate, than tested environmental conditions.