Genetic engineering in the improvement of plants for phytoremediation of metal polluted soils

Metal concentrations in soils are locally quite high, and are still increasing due to many human activities, leading to elevated risk for health and the environment. Phytoremediation may offer a viable solution to this problem, and the approach is gaining increasing interest. Improvement of plants by genetic engineering, i.e. by modifying characteristics like metal uptake, transport and accumulation as well as metal tolerance, opens up new possibilities for phytoremediation. So far, only a few cases have been reported where one or more of these characteristics have been successfully altered; e.g. mercuric ion reduction causing improved resistance and phytoextraction, and metallothionein causing enhanced cadmium tolerance. These, together with other approaches and potentially promising genes for transformation of target plants are discussed.     

Cu accumulation in the earthworm Dendrobaena veneta in a heavy metal (Cu, Pb, Zn) contaminated site compared to Cu accumulation in laboratory experiments

Earthworms (Dendrobaena veneta) were exposed to heavy metal contaminated soil from a field site under both laboratory and field conditions. In the laboratory study, D. veneta were analysed for Cu after 2 weeks' exposure. The tissue Cu concentration (Cu(W)) increased proportionally with the total extractable soil Cu content (Cu(T)) in soils that contained less than 150 mg Cu kg(-1). In D. veneta exposed to soils containing more than Cu(T) = 150 mg kg(-1), however, we observed no further increase in Cu(W). In a 64-m(2) field plot at a contaminated site, 81 soil samples were taken and analysed for Cu, Pb, Zn and pH. In June 1994, 1000 D. veneta were homogeneously distributed in the plot and sampled after 1, 2 and 5 weeks to analyse for Cu. Additionally, 2000 D. veneta were introduced in September 1994 and sampled after 1, 2 and 4 weeks. At locations where earthworms were removed, the soil Cu content was estimated by disjunctive kriging. Cu concentration factors (ratio of Cu(W) to Cu(T)) in the field experiment were in excellent agreement with those of the laboratory experiment. This indicates that, with a sufficiently high sampling density, it may be feasible to predict earthworm Cu accumulation in the field using a concentration factor determined in the laboratory.     

The influence of time on lead toxicity and bioaccumulation determined by the OECD earthworm toxicity test

Internationally agreed standard protocols for assessing chemical toxicity of contaminants in soil to worms assume that the test soil does not need to equilibrate with the chemical to be tested prior to the addition of the test organisms and that the chemical will exert any toxic effect upon the test organism within 28 days. Three experiments were carried out to investigate these assumptions. The first experiment was a standard toxicity test where lead nitrate was added to a soil in solution to give a range of concentrations. The mortality of the worms and the concentration of lead in the survivors were determined. The LC50s for 14 and 28 days were 5311 and 5395 microgPb g(-1)soil respectively. The second experiment was a timed lead accumulation study with worms cultivated in soil containing either 3000 or 5000 microgPb g(-1)soil. The concentration of lead in the worms was determined at various sampling times. Uptake at both concentrations was linear with time. Worms in the 5000 microg g(-1) soil accumulated lead at a faster rate (3.16 microg Pb g(-1)tissue day(-1)) than those in the 3000 microg g(-1) soil (2.21 microg Pb g(-1)tissue day(-1)). The third experiment was a timed experiment with worms cultivated in soil containing 7000 microgPb g(-1)soil. Soil and lead nitrate solution were mixed and stored at 20 degrees C. Worms were added at various times over a 35-day period. The time to death increased from 23 h, when worms were added directly after the lead was added to the soil, to 67 h when worms were added after the soil had equilibrated with the lead for 35 days. In artificially Pb-amended soils the worms accumulate Pb over the duration of their exposure to the Pb. Thus time limited toxicity tests may be terminated before worm body load has reached a toxic level. This could result in under-estimates of the toxicity of Pb to worms. As the equilibration time of artificially amended Pb-bearing soils increases the bioavailability of Pb decreases. Thus addition of worms shortly after addition of Pb to soils may result in the over-estimate of Pb toxicity to worms. The current OECD acute worm toxicity test fails to take these two phenomena into account thereby reducing the environmental relevance of the contaminant toxicities it is used to calculate.     

Effect of formaldehyde on biofilm activity and morphology in an ultracompact biofilm reactor for carbonaceous wastewater treatment.

The effects of formaldehyde on biofilm morphology and biomass activity were investigated in an ultracompact biofilm reactor (UCBR) for carbonaceous wastewater treatment. The wastewater contained a fixed amount of glucose (with a chemical oxygen demand concentration of 600 mg/L) and an increasing concentration of formaldehyde (ranging from 21.4 to 271.1 mg/L). An influent formaldehyde concentration higher than 75 mg/L could facilitate filamentous growth (on biofilm) control and lead to a higher biofilm density, which is desirable as it enhanced the UCBR performance stability. However, at an influent formaldehyde concentration higher than 214.4 mg/L, biomass production was inhibited and deteriorations of biofilm morphology and biomass activity were observed. This study showed that it was desirable to maintain an influent formaldehyde concentration lower than 202.2 mg/L, as this concentration could achieve a good biofilm morphology while not inhibiting its microbial activity.     

Potential role of fire retardant-treated polyurethane foam as a source of brominated diphenyl ethers to the US environment

Five tetra- to hexabrominated diphenyl ether (BDE) congeners (BDE-47, -99, -100, -153 and -154) are the most frequently reported in wildlife and humans. The commercial penta-BDE product, used predominantly to flame-retard polyurethane foam, consists primarily of these same congeners. In 1999, North American demand accounted for 98% of the total global penta-market of 8500 metric tons. Frogs, housed with flame retardant-treated polyurethane foam as a dry substrate, accumulated 10,100 microg/kg (wet weight) of the above BDEs. Crickets kept therein as food contained 14,400 microg/kg. The crickets are believed to have browsed directly on the foam and, in turn, were consumed by the frogs. BDE congener composition in all three matrices matched that of the penta-commercial product. Similar congeners were also observed in soil and stream sediments collected near a polyurethane foam manufacturing plant. Summed concentrations of BDE-47, -99 and -100, the dominant congeners observed in these samples, ranged from < 1 to 132 microg/kg (dry weight basis). Sunfish fillets obtained from a nearby, off-site pond contained a total of 624 microg/kg (lipid basis). Sewage treatment plant (STP) sludge exhibited these same congeners at 1370 microg/kg (dry weight). BDE-209, the fully brominated congener predominant in the commercial deca-BDE product, was also present at 1470 microg/kg. While no known polyurethane foam manufacturers discharged to this plant, the distribution pattern of the low brominated congeners in the sludge matched that of the penta-product. After four weeks of exposure to ambient outdoor conditions, the surface of flame-retarded polyurethane foam became brittle and began to disintegrate. Subsequent dispersal of these penta-containing foam fragments may be one mechanism by which these BDEs reach the environment.     

Individual PCBs as predictors for concentrations of non and mono-ortho PCBs in human milk

32 Dutch human milk samples were analyzed for PCBs with either HRGC-ECD or HRGC-LRMS in the NCI mode. Samples were collected from three different locations in The Netherlands: Amsterdam, Rotterdam and Groningen. Quantitatively, no differences could be observed between the three localities, while in addition the congener specific pattern showed a striking similarity for all individual samples. Only principal component analysis revealed slight individual differences. Based on similarities in the PCB profiles, linear relationships were calculated between 2,3′4,4′,5-PnCB (#118) or 2,2′4,4′5,5′HxCB (#153) and the most relevantnon andmonoortho PCBs exhibiting dioxinlike activity. These PCBs included 2,3,3′,4,4′-PnCB (#105), 3,3′,4,4′5-PnCB (#126) 2,3,3′,4,4′,5-HxCB (#156), 2,3,3′,4,4′,5′-HxCB (#157), 2,3′,4,4′,5,5′-HxCB (#167) and 3,3′,4,4′,5′5-HxCB (#169). Good linear relationships were observed between individual PCBs. Based on the results of this study, PCB #118 can be used to predict concentrations of the PCBs #105 and #126. PCB #153 can be used as a predictor for the PCBs #156, #157, #167 and #169, but also for the total toxic equivalencies (TEQs) ofnon andmonoortho PCBs present in human milk. This method using certain PCBs as predictors for other toxicological relevant congeners, can be useful and cost effective, e.g. for epidemiological studies. However, before applied a number of conditions should be met. These are:

1. A stable composition of the PCB matrix should be established.
2. A possible time dependent change in composition of the matrix should first be excluded when used over different time periods.

     

Intercomparison of reactive transport models applied to UO2 oxidative dissolution and uranium migration

Oxidative dissolution of uranium dioxide (UO₂) and the subsequent migration of uranium in a subsurface environment and an underground waste disposal have been simulated with reactive transport models. In these systems, hydrogeological and chemical processes are closely entangled and their interdependency has been analyzed in detail, notably with respect to redox reactions, kinetics of mineralogical evolution and hydrodynamic migration of species of interest.Different codes, where among CASTEM, CHEMTRAP and HYTEC, have been used as an intercomparison and verification exercise. Although the agreement between codes is satisfactory, it is shown that the discretization method of the transport equation (i.e. finite elements (FE) versus mixed-hybrid FE and finite differences) and the sequential coupling scheme may lead to systematic discrepancies.     

Solvent and surfactant enhanced solubilization, stabilization, and degradation of amitraz

In an effort to help with the development of effective dip vat management and waste disposal strategies this study determined how solution properties such as pH, buffer composition, ionic strength, temperature, solubility in organic solvents and the addition of commonly used solubilizing agents influenced the hydrolysis of amitraz. Amitraz degrade by means of hydrolysis described by a pseudo-first order rate process and a type ABCD pH rate profile. Hydrolysis increased with temperature and was fastest at low pH, slowest at neutral to slightly alkaline pH, and slightly increased above pH 10. However, buffer concentration and ionic strength influenced the hydrolysis rate and had to be accounted for before constructing a pH rate profile. Hydrolysis seems to depend on the dielectric constant of solvent mixtures and was fastest in water, slower in propylene glycol and ethanol solutions, and slowest in DMSO mixtures. In surfactant solutions, anionic micelles enhanced and cationic micelles retarded the hydrolysis rate. The magnitude of micellar effects decreased with increasing concentrations of the surfactants. The increased solubility and faster hydrolysis of amitraz in the sodium lauryl sulfate solutions showed that anionic surfactants potentially could be used for cleaning up amitraz spills, because it both solubilized the drug and catalyzed hydrolysis.