The comparison of rapid bioassays for the assessment of urban groundwater quality

Groundwater is a complex mixture of chemicals that is naturally variable. Current legislation in the UK requires that groundwater quality and the degree of contamination are assessed using chemical methods. Such methods do not consider the synergistic or antagonistic interactions that may affect the bioavailability and toxicity of pollutants in the environment. Bioassays are a method for assessing the toxic impact of whole groundwater samples on the environment. Three rapid bioassays, Eclox, Microtox and ToxAlert, and a Daphnia magna 48-h immobilisation test were used to assess groundwater quality from sites with a wide range of historical uses. Eclox responses indicated that the test was very sensitive to changes in groundwater chemistry; 77% of the results had a percentage inhibition greater than 90%. ToxAlert, although suitable for monitoring changes in water quality under laboratory conditions, produced highly variable results due to fluctuations in temperature and the chemical composition of the samples. Microtox produced replicable results that correlated with those from D. magna tests.     

Impact of Bioaugmentation with a Consortium of Bacteria on the Remediation of Wastewater-Containing Hydrocarbons (5 pp)

Goals, Scope and Background It has been observed that hydrocarbon treated wastewaters still contain high COD and a number of intermediates. This suggests that the required catabolic gene pool for further degradation might be absent in the system or, that its titer value is not significant enough. By providing the desired catabolic potential, the overall efficiency of the treatment system can be improved. This study aims to demonstrate this concept by bioaugmentation of a lab-scale reactor treating refinery wastewater with a consortium having the capacity to complement the alkB genotype to the available microbial population. Methods Two reactors were set up using activated biomass collected from a refinery treatment plant and operated at a continuous mode for a period of 8 weeks. The feed to both reactors was kept constant. Crude oil was spiked regularly. One reactor was bioaugmented with a consortium previously described for crude oil spill remediation. The efficiency of the bioaugmented reactor was demonstrated by reduced COD. The changes in the microbial population over a period of time were analyzed by RAPD. Catabolic activity of the biomass in both reactors was monitored by PCR. The presence of the catabolic loci was confirmed by Southern Hybridization. Results and Discussion 52.2% removal of COD was observed in the bioaugmented reactor while only 15.1% reduction of COD was observed in the reactor without bioaugmentation. The change in microbial population can be seen from the 4th week, which also corresponds to improved catabolic activity. The presence of the bedA locus was seen in all samples, which indicates the presence of aromatic degraders, but the appearance of the alkB locus, from the 6th week onwards, which was observed only in the samples from the bioaugmented reactor. The results suggest that the gene pool of the bioaugmented reactor has catabolic loci that can degrade accumulated intermediates, thus improving the efficiency of the system. Conclusions In this study, improvement of efficiency of bioremediation was demonstrated by addition of catabolic loci that are responsible for degradation. Bioaugmentation was carried out in biomass that was collected from an ETP (effluent treatment plant) treating hydrocarbon containing wastewater to study the strategies for improvement of the treatment system. Biostimulation, only marginally improved the efficiency, when compared to bioaugmentation. The improved efficiency was demonstrated by COD removal. The presence of the alkB locus suggests the importance of a catabolic gene pool that acts on accumulated intermediates. It is well documented that straight chain aliphatics and intermediates of aromatic compounds after ring cleavage, accumulate in refinery wastewater systems, thereby hindering further degradation of the wastewater. Supplementation of a catabolic gene pool that treats the lower pathway compounds and alkanes will improve the overall efficiency. In this study, results suggest that the alkB locus can also be used to monitor the degradative mode of the activated biomass. Recommendations and Perspective . Pollution from petroleum and petroleum products around the globe are known to have grave consequences on the environment. Bioremediation, using activated sludge, is one option for the treatment of such wastes. Effluent treatment plants are usually unable to completely degrade the wastewater being treated in the biological unit (the aerator chambers). The efficiency of degradation can be improved by biostimulation and bioaugmentation. This study demonstrates the improved efficiency of a treatment system for wastewater containing hydrocarbons by bioaugmentation of a consortium that supports degradation. Further experiments on a pilot scale are recommended to assess the use of bioaugmentation on a large scale. The use of molecular tools, like DNA probes for alkB, to monitor the system also needs to be explored.     

Modification of Langmuir isotherm in solution systems--definition and utilization of concentration dependent factor

The Langmuir isotherm, originally derived for the adsorption of gas molecules on solid surfaces, was modified to fit the adsorption isotherm of solutes onto solid surfaces in solution systems. The aim of this modification is based on the fact that direct application of the Langmuir isotherm to solution systems often leads to poor data fitting. In the present communication, it is shown that the level of data fitting to the Langmuir isotherm of literature data can be improved by a simple modification introducing a concentration dependent factor, X. The key concept of the modification lies in that the concentration of solute affects both adsorption and desorption stages. As a first approximation, we adopted a single-term polynomial for both processes of adsorption and desorption. Based on reanalysis of literature data of adsorption in solution, we confirmed that indeed the modified Langmuir isotherm more accurately describes the experimental observations. Furthermore, we proposed that the concentration dependent factor could be associated with the surface heterogeneity index that was introduced in a few other modified Langmuir isotherms. Some advantages and limitations of proposed modified Langmuir isotherm are also discussed.     

Use of tree rings to investigate the onset of contamination of a shallow aquifer by chlorinated hydrocarbons

Oaks (Quercus velutina Lam.) growing over a shallow aquifer contaminated by chlorinated hydrocarbons were studied to determine if it was possible to estimate the approximate year that contamination began. The annual rings of some trees downgradient from the contaminant release site contained elevated concentrations of chloride possibly derived from dechlorination of contaminants. Additionally, a radial-growth decline began in these trees at approximately the same time that chloride became elevated. Growth did not decline in trees that contained smaller concentrations of chloride. The source of elevated chloride and the corresponding reductions in tree growth could not be explained by factors other than contamination. On the basis of tree-ring evidence alone, the release occurred in the late 1960s or early 1970s. Contaminant release at a second location apparently occurred in the mid- to late 1970s, suggesting that the area was used for disposal for at least 5 years and possibly longer.     

Assessment of bioconcentration and secondary poisoning of surfactants

The relevance of the bioconcentration behaviour of surfactants for the secondary poisoning assessment and for the risk characterisation in the bird and mammalian food chain has been investigated. The approach used is described in the recently revised EU Technical Guidance Document for the Risk Assessment of Substances. The results demonstrate that, based on experimentally derived bioconcentration factors, environmental concentrations and effects in animals, there is a clear level of safety for both linear alkylbenzene sulphonate (LAS) and alcohol ethoxylates (AE), the most important surfactants by volume. To assess other surfactants used in detergents, a bioconcentration factor that would need to be attained for secondary poisoning to be of concern has been estimated from predicted environmental concentrations and known long-term effects data in animals. Based on the known structural similarity of these surfactants to LAS and AE and the ubiquitous nature of the enzymatic systems that are present in biotransformation processes in organisms, it is concluded that bioconcentration of these surfactants to these levels is highly unlikely. Therefore the potential for secondary poisoning effects of these surfactants is extremely low.     

A spectroscopic study of the bromination of the endocrine disruptor ethynylestradiol

Bromination of ethynylestradiol (EE2) was studied using fluorescence and conventional and stopped-flow absorbance spectroscopy. EE2 transformations in the presence of bromine were determined to proceed through sequential reactions. The first step is very rapid and results in the formation of a brominated cyclohexadienone that is susceptible to reduction but in oxidative conditions it undergoes a transformation into monobromo-EE2 in a first-order OH- -accelerated reaction. The transformation from monobromo-EE2 to dibromo-EE2 follows a similar mechanism. The rates of reactions of bromine-substituted EE2 species are orders of magnitude lower than that of the parent compound indicating that these species can persist in conditions typical for water treatment operations.     

Evaluation of the exposure methodology for risk-benefit assessment of seafood consumption

This study evaluated the methodology of using existing, international data to calculate the intake of nutrients and contaminants via seafood consumption on a national level. This was done by combining French seafood consumption data with two different datasets. The results showed that a significant correlation was found between the results obtained by using different data sets and that a high percentage of the population was classified in the same category (above or below the intake reference values). As such, the general conclusion that can be drawn from the results: 'whether a part of the population does exceed the toxicological reference values for certain contaminants and whether a part of the population does not reach the recommended intake for certain nutrients' remains the same for the two approaches. Therefore, it can be concluded that the existing, international probability distributions are useful when aiming to perform a cost-effective risk benefit evaluation of seafood consumption on national or regional level. When this evaluation does indicate that more detailed results are needed because a certain health risk exists, it can be decided that it is worth to spend more time and money for the analyses of national samples in order to collect more detailed results.     

Source and persistence of pesticides in a semi-confined chalk aquifer of southeast England

Pesticide contamination in groundwater is an increasing problem that poses a significant long-term threat to water quality. Following the detection of elevated concentrations of diuron in boreholes in a semi-confined chalk aquifer from southeast England, a sampling programme was undertaken. Between 2003 and 2004 diuron was observed in 90% of groundwaters analysed. In 60% of groundwater samples metabolites of diuron were more prevalent than the parent compound. Longer-term (1989-2005) monitoring shows that pollution of the aquifer by atrazine, simazine, and more recently diuron, shows a positive correlation with periods of high groundwater levels. Results from groundwater residence time indicators suggest that the highest diuron concentrations are associated with waters containing the greatest proportion of recent recharge. There is some evidence to indicate that diuron occurrence can be spatially related to areas of urban and industrial development and is probably correlated with amenity usage.     

Variability of dust iron solubility in atmospheric waters: Investigation of the role of oxalate organic complexation

Atmospheric dust deposition is a major external iron source for remote surface ocean waters. Organic complexation is known to play a role in the dissolution of iron-containing minerals. In this paper, we present our study on the effect of oxalate on dust iron solubility in simulated rainwater. Our results reveal that the solubility of iron carried by analogs of different African dusts varies with the dust source. Our experiments indicate a positive linear correlation between iron solubility and oxalate concentration. Soluble iron (SFe) increases from 0.0025(±0.0005)% to 0.26(±0.01)% of total iron, considering all dust sources and with oxalate concentrations ranging from 0 to 8 μM. These results show that the observed variability of iron solubility in aerosols collected over the Atlantic Ocean is, at least partly, due to an increase in dust iron solubility, with the presence of oxalate complexation, rather than to the presence of more soluble anthropogenic iron. Considering the mineralogical composition of those particles, experiments with pure minerals (hematite, goethite and illite) were performed to study the dissolution process. We found that oxalate promotes the solubility of iron contained in clay and hence confirmed that more than 95% of SFe from soil dust is provided by clay (illite). This experimental work enables us to establish a parameterization of iron solubility in dust as a function of oxalate concentration and based on the individual iron solubility of pure iron-bearing minerals usually present in dust particles. Finally, our results emphasize that oxalate contributes to iron solubility on the same order of magnitude as the acid processes. Organic complexation appears to be a process that increases iron solubility and likely enhances the bioavailability of iron from dust.     

Modifications of the sex pheromonal communication of trichogramma brassicae by a sublethal dose of deltamethrin

Pyrethroid insecticides, by their intensive use and their persistence that can exceed 3 months, largely contribute to the environmental pollution. In this work, we determined the effects of a very low dose of deltamethrin on the sex pheromonal communication of Trichogramma brassicae. The dose used was a dose that would theoretically kill one insect over 1000 (an LD 0.1). We found that this dose slightly but very significantly increased the arrestment behavior of treated males responding to the female pheromone. On the other hand when females were exposed to the same dose of insecticide, the response of males to their pheromone was very significantly decreased. In Trichogramma, like in other insects, sex pheromonal communication probably involves nervous transmissions both for the reception and the emission of the pheromone. Then, the sublethal effects of deltamethrin, observed in this work, are certainly due to multiple actions of this insecticide on nervous transmissions. Trichogramma is a beneficial insect that contributes to the control of pest populations of moths. Actions of this insecticide at a dose that can correspond to environmental pollution could be a real threat to the equilibrium of these populations.     

Assessing the risk of impact of farming intensification on calcareous grasslands in Europe: a quantitative implementation of the MIRABEL framework

Intensification of farming practices is still a major driver of biodiversity loss in Europe, despite the implementation of policies that aim to reverse this trend. A conceptual framework called MIRABEL was previously developed that enabled a qualitative and expert-based assessment of the impact of agricultural intensification on ecologically valuable habitats. We present a quantitative update of the previous assessment that uses newly available pan-European spatially explicit data on pressures and habitats at risk. This quantitative assessment shows that the number of calcareous grasslands potentially at risk of eutrophication and overgrazing is rapidly increasing in Europe. Decreases in nitrogen surpluses and stocking densities that occurred between 1990 and 2000 have rarely led to values that were below the ecological thresholds. At the same time, a substantial proportion of calcareous grassland that has so far experienced low values for indicators of farming intensification has faced increases between 1990 and 2000 and could well become at high risk from farming intensification in the near future. As such, this assessment is an early warning signal, especially for habitats located in areas that have traditionally been farmed extensively. When comparing the outcome of this assessment with the previous qualitative MIRABEL assessment, it appears that if pan-European data are useful to assess the intensity of the pressures, more work is needed to identify regional variations in the response of biodiversity to such pressures. This is where a qualitative approach based on regional expertise should be used to complement data-driven assessments.     

Economic Impact of Mass Production of Alternative Low Emission Automotive Power Systems

The study considers a range of possible effects on the transportation industry, satellite industries, the labor market, and the economy which may be anticipated in the event mass production of unconventional low emission automotive propulsion systems should occur, whether as a consequence of federal intervention, or not. A postulated 1 976 Otto Cycle Internal Combustion Engine equipped with a dual catalyst manifold reactor and other “conventional” emission control devices was compared in detail with a Regenerative Free Turbine Engine and a Rankine Cycle Engine, as specified by the contracting agency. Manufacturing costs, operating and ownership costs, consumer demand, inter-industry effects, employment, resource requirements, and international trade implications were analyzed in depth under a number of plausible sets of policy constraints and parametric variations. Principal conclusions are that conversion over a 10 year period is feasible, that manufacturing cost differentials are less critical than fuel consumption and cost differentials, that industry/employment impacts are minor, and that resource/trade effects are dominated by petroleum imports. Implications for federal policy are discussed.     

The fate of some components of acidic deposition in ombrotrophic mires

The ability of ombrotrophic Sphagnum species to immobilise inorganic nitrogen deposited from the atmosphere was investigated in a series of simple lysimeter experiments. In an unpolluted mire, Sphagnum fuscum mats retained all the nitrogen deposited to them in natural precipitation events. Sphagnum capillifolium mats, transplanted from an unpolluted site to the polluted southern Pennines in England, also initially retained a large proportion of deposited nitrate and ammonium. However, a laboratory experiment demonstrated that high rates of nitrogen supply cause a loss of the ability of the moss to retain nitrate, which suggests that this may occur as a result of increased nitrogen deposition in polluted regions, resulting in increased nitrate availability in the peat. Investigation of the volume of precipitation and amounts of sulphate and chloride passing through the Sphagnum mats in the southern Pennines, as compared to that collected in adjacent bulk deposition gauges, showed that conventional deposition monitoring grossly underestimates rates of deposition to vegetation. Efficient trapping of occult and dry deposition by the moss led to much greater volumes of precipitation and amounts of sulphate and chloride being measured in throughflow than in bulk precipitation samples. Physiological response of S. fuscum to occult precipitation and heavy rainfall was investigated by measuring nitrate reductase activity induced in the moss by nitrate supplied in 'fine mist' and 'large droplet' applications of solutions to moss in the field. Greater response was shown to occult deposition, suggesting that this form of precipitation may be important in vegetation damage in polluted regions.     

Distribution of heavy metal contents of urban soils in parks of Seville

Several metals in urban soils of Seville were extracted by two methods, which respectively give estimates of the available and ‘quasi total’ contents. Although the soils did not show strong heterogeneity in their general properties, high dispersion of the contents in those metals with greater relative availability, Cu, Pb and Zn, as compared to others suggested that pollution with these three metals could occur in some sampling sites. It was shown that the contents in these metals tend to increase as the historic quarters of the city are approached. Using reference values given by the Québec Ministry of Environment it was shown that those green areas closer to the historic centre present contents in Pb, Zn and particularly Cu that often exceed the acceptable limits for residential, recreational and institutional sites. From background contents for Seville soils estimated from a park located on the outskirts, a pollution load index (PLI) for each sampling site was calculated for the set of these three metals. It was shown that the PLI tends to increase as traffic density increases and as distance from main traffic decreases, but poor regressions were obtained, suggesting that other variables different from traffic should be considered.     

Degradation of para-hydroxybenzoic acid by means of mixed microbial cultures

Olive mill wastewater contains some phenolic compounds that cause antibacterial activity of a kind that prevents biological treatment without previous dilution. Among these phenolic compounds, p-hydroxybenzoic acid (PHB) is considered to be one of the most representative. This work examines the biodegradation of PHB by aerobic microbial mixed cultures previously acclimatized to glucose, which was used as an easily biodegradable model compound. Microbial growth runs were carried out in a batch reactor in the PHB concentration range of 200-1000 mg/L. In all the runs the PHB proved to be completely degradable. The specific growth rates obtained were in the range of 0.16-0.35 l/h. Experimental runs showed that the functional relationship between the specific growth rate and PHB concentration was that proposed by Monod. The kinetic constants of the Monod equation (mu(max) and K(S)) and biomass yield coefficient (Y) were determined experimentally. With the parameter values thus obtained, a mathematical model that also takes account of the duration of the lag phase was employed to describe both the microbial growth and the consumption of PHB. The concentration values of the model fit well with the data obtained experimentally.     

No enhancement in bioconcentration of organic contaminants by low levels of DOM.

The aim of the present work was to systematically study the effect of low concentrations of dissolved organic matter (DOM) on the bioconcentration of organic contaminants, in order to show whether the phenomenon of enhanced bioconcentration factors (BCFs), that has been reported in the literature, is generally found at low levels of DOM or if BCF enhancements are more likely due to a random scatter in the experimental data. The first part of the study tested the hypothesis that low levels of DOM affect the uptake kinetics of organic contaminants, leading to transient enhancements of BCFs, relative to DOM-free controls, which could have been reported as BCF enhancements in short-term studies. We found that the presence of low concentrations of two different types of DOM consistently decreased the bioconcentration of benzo[a]pyrene (BaP) in the water flea Daphnia magna at all exposure times (1-24 h), and that no transient BCF enhancements occurred. The second part of the study systematically investigated if low concentrations of DOM from a wide range of different aquatic systems can cause enhancements in the bioconcentration of organic contaminants. Water fleas were exposed to combinations of four different organic contaminants (BaP, tetrachlorobiphenyl, pentachlorophenol and naphthalene) with low concentrations of 12 different types of DOM that had been collected from various regions throughout Europe. In several of the DOM treatments, we found mean BCFs being higher than mean BCFs in the controls (especially for naphthalene). This shows that the experimental setup used in this study (and similarly in previous studies) can produce seeming BCF enhancements at low concentrations of DOM. However, statistical analyses showed that treatment means were not significantly different from control means. Thus, this systematic study suggests that the BCF enhancements that have been reported in the literature are more likely the result of random, experimental variations than the result of a systematic enhancement of bioconcentration.     

The impact of additives found in industrial formulations of TCE on the wettability of sandstone

The wettability of aquifer rocks is a key physical parameter which exerts an important control on the transport, residual trapping, distribution and eventual fate of chlorinated hydrocarbon solvents (CHSs) released into the subsurface. Typically chlorinated solvents are assumed to be non-wetting in water saturated rocks and unconsolidated sediments. However industrially formulated solvent products are often combined with basic additives such as alkylamines to improve their performance; and the mineral surfaces of aquifer rocks and sediments usually possess a range of acid and hydrogen-bonding adsorption sites. The presence of these sites provides a mechanism whereby the basic additives in CHSs can be adsorbed at the solvent phase/solid phase interface. Given the amphiphilic molecular structure of these additives, this may result in changes in the wetting conditions of the solid phase. The aim of this study was therefore to test this conjecture for two classes of additives (alkylamines and quaternary ammonium salts) that are often encountered in industrial solvent formulations. Wettability assessments were made on sandstone cores by means of measurements of spontaneous and forced water drainage and spontaneous and forced water imbibition and through contact angle measurements on a smooth quartz surface. No solvent/additive combination produced solvent wetting conditions, though dodecylamine and octadecylamine significantly reduced the water wetting preference of sandstone which frequently resulted in neutral wetting conditions. The large volume of spontaneous water drainage observed in wettability experiments involving cetyltrimethylammonium bromide and octadecyltrimethylammonium bromide, suggested that the sandstone cores in these tests remained strongly water wetting. However equilibrium static contact angles of around 60 degrees were measured on quartz suggesting that the sandstone surfaces should be close to neutral wetting conditions. This paradox was finally resolved by noting that contact between the solvent mixture and water in the sandstone core resulted in a final solvent phase which had an extremely low interfacial tension. It is therefore suspected that the observed spontaneous drainage of solvent from the core was driven by gravitational and buoyancy forces rather than strong water wetting conditions. Finally it was noted that the mobilisation of iron oxide coatings from the sandstone surface had a considerable influence in reducing the interfacial tension and in the formation and stabilisation of TCE/water emulsions.     

Optimization of Height of Release of Air Pollutants

A number of key projects in the Environmental Protection Agency (EPA) particulate R&;D program having applicability to industry are presented. For electrostatic precipitators (ESP) there is presented the result of work on large diameter discharge electrodes which provide a decrease in penetration of up to a factor of 4 when compared to conventional small diameter electrodes. Also discussed is the multistage ESP which provides a collection efficiency that would require a collecting plate area 4 or 5 times larger with conventional ESP technology. The E-SOX technology makes use of the multistage concept to free up space in the ESP for SO₂ removals of up to 90%. Electrostatically augmented fabric filtration provides a reduction in pressure drop of about 5 0% as compared to conventional fabric filtration. Wind tunnel modeling of windbreaks for material storage piles indicates a potential for providing engineering design data that would allow significant emission reduction caused by wind erosion     

A study of 2,4,6-trinitrotoluene inhibition of benzo[a]pyrene uptake and activation in a microbial mutagenicity assay

A number of in vitro and in vivo studies have determined that binary and complex mixtures may interact to produce a toxicity that could not be predicted based on the individual chemicals. The present study was conducted with a binary mixture of model compounds to investigate possible interactions affecting their mutagenicity. The compounds included Benzo[a]pyrene (BAP), a polycyclic aromatic hydrocarbon that is an indirect-acting mutagen of great environmental concern, and 2,4,6-Trinitrotoluene (TNT), a nitro-aromatic compound that is a direct-acting mutagen frequently found as a soil contaminant at munitions sites. This study indicated that a binary mixture of BAP and TNT failed to induce the positive mutagenic response in Salmonella typhimurium strain TA98 characteristic of either compound alone. Spectrofluorometric analysis of BAP, and kinetic analyses of ³HBAP uptake in the presence or absence of TNT using TA98 cells that were treated or untreated with activated rat liver microsomes were performed. In cells preloaded with BAP, cellular BAP fluorescence was rapidly suppressed in the presence of TNT. Mass spectroscopy of BAP and TNT mixtures revealed a number of products, believed to be the result of complexation and nitration, that may account for the antagonistic action of TNT on BAP-induced mutagenicity in TA98 cells. Further, kinetic studies indicated that TNT inhibited the incorporation of BAP into cells.     

Model-based experimental design for assessing effects of mixtures of chemicals

We exposed flour beetles (Tribolium castaneum) to a mixture of four poly aromatic hydrocarbons (PAHs). The experimental setup was chosen such that the emphasis was on assessing partial effects. We interpreted the effects of the mixture by a process-based model, with a threshold concentration for effects on survival. The behavior of the threshold concentration was one of the key features of this research. We showed that the threshold concentration is shared by toxicants with the same mode of action, which gives a mechanistic explanation for the observation that toxic effects in mixtures may occur in concentration ranges where the individual components do not show effects. Our approach gives reliable predictions of partial effects on survival and allows for a reduction of experimental effort in assessing effects of mixtures, extrapolations to other mixtures, other points in time, or in a wider perspective to other organisms.