Toxic effects of chemical warfare agent mixtures on the mussel Mytilus trossulus in the Baltic Sea: A laboratory exposure study.

Baltic blue mussels (Mytilus trossulus) were implemented to assess potential toxicity, health impairments and bioaccumulation of dumped chemical warfare agents on marine benthic organisms. Mussels were collected from a pristine cultivation side and exposed under laboratory conditions to different mixtures of chemical warfare agents (CWAs) related phenyl arsenic compounds, Clark I and Adamsite as well as chloroacetophenone. Using a multi-biomarker approach, mussels were assessed thereafter for effects at different organisational levels ranging from geno-to cytotoxic effects, differences in enzyme kinetics and immunological responses. In an integrated approach, chemical analysis of water and tissue of the test organisms was performed in parallel. The results show clearly that exposed mussels bioaccumulate the oxidized forms of chemical warfare agents Clark I, Adamsite (DAox and DMox) and, to a certain extent, also chloroacetophenone into their tissues. Adverse effects in the test organisms at subcellular and functional level, including cytotoxic, immunotoxic and oxidative stress effects were visible. These acute effects occurred even at the lowest test concentration.

Toxicity and accumulation of trinitrotoluene (TNT) and its metabolites in Atlantic salmon alevins exposed to an industrially polluted water.

A pond in an industrial area in Sweden was selected to study adverse effects on salmon alevins from 2,4,6-trinitrotoluene (TNT)-contaminated water. Chemical screening revealed heavy contamination of TNT and its degradation products, 2-amino-4,6-dinitrotoluene (2-ADNT) and 4-amino-2,6-dinitrotoluene (4-ADNT), ranging from 0.05 to 230 g/kg in the sediment (dry weight) within the water system. Pond water contained 3 mg/L TNT. A dilution series of pond water mixed with tap water revealed increased death frequency in alevins down to fivefold dilution (approximate 0.4 mg TNT/L). Uptake was concentration dependent, reaching 7, 9, and 22 µg/g tissue for TNT, 2-ADNT, and 4-ADNT at the highest test concentration. A time-dependent uptake of TNT and its degradation products was found at a water concentration of 0.08 mg TNT/L. Degradation products of TNT showed a more efficient uptake compared to native TNT, and accumulation of 4-ADNT was more pronounced during the late phase of the 40-d exposure study. Bioconcentration factors (BCF) (0.34, 52, and 134 ml/g for TNT, 2-ADNT, and 4-ADNT, respectively) demonstrated a significant uptake of the metabolite 4-ADNT in alevin tissue. Disturbed physiological conditions and delayed development in alevins were not studied, but may not be excluded even at 125-fold diluted pond water (0.016 mg TNT/L). BCF data indicated that bioaccumulation of TNT metabolites need to be considered in TNT chronic toxicity. Fish species and age differences in the accumulation of TNT metabolites need to be further studied.

The effect of subsurface military detonations on vadose zone hydraulic conductivity, contaminant transport and aquifer recharge.

Live fire military training involves the detonation of explosive warheads on training ranges. The purpose of this experiment is to evaluate the hydrogeological changes to the vadose zone caused by military training with high explosive ammunition. In particular, this study investigates artillery ammunition which penetrates underground prior to exploding, either by design or by defective fuze mechanisms. A 105 mm artillery round was detonated 2.6 m underground, and hydraulic conductivity measurements were taken before and after the explosion. A total of 114 hydraulic conductivity measurements were obtained within a radius of 3m from the detonation point, at four different depths and at three different time periods separated by 18months. This data was used to produce a three dimensional numerical model of the soil affected by the exploding artillery round. This model was then used to investigate potential changes to aquifer recharge and contaminant transport caused by the detonating round. The results indicate that an exploding artillery round can strongly affect the hydraulic conductivity in the vadose zone, increasing it locally by over an order of magnitude. These variations, however, appear to cause relatively small changes to both local groundwater recharge and contaminant transport.

Ecotoxicological assessment and evaluation of a pine bark biosorbent treatment of five landfill leachates.

When selecting a landfill leachate treatment method the contaminant composition of the leachate should be considered in order to obtain the most cost-effective treatment option. In this study the filter material pine bark was evaluated as a treatment for five landfill leachates originating from different cells of the same landfill in Sweden. The objective of the study was to determine the uptake, or release, of metals and dissolved organic carbon (DOC) during a leaching test using the pine bark filter material with the five different landfill leachates. Furthermore the change of toxicity after treatment was studied using a battery of aquatic bioassays assessing luminescent bacteria (Vibrio fischeri) acute toxicity (30-min Microtox®), immobility of the crustacean Daphnia magna, growth inhibition of the algae Pseudokirchneriella subcapitata and the aquatic plant Lemna minor; and genotoxicity with the bacterial Umu-C assay. The results from the toxicity tests and the chemical analysis were analyzed in a Principal Component Analysis and the toxicity of the samples before and after treatment was evaluated in a toxicity classification. The pine bark filter material reduced the concentrations of metal contaminants from the landfill leachates in the study, with some exceptions for Cu and Cd. The Zn uptake of the filter was high for heavily contaminated leachates (≥73%), although some desorption of zinc occurred in less contaminated waters. Some of the leachates may require further treatment due to discharge into a natural recipient in order to reduce the risk of possible biological effects. The difference in pH changes between the different leachates was probably due to variations in buffering capacity, affected by physicochemical properties of the leachate. The greatest desorption of phenol during filtration occurred in leachates with high conductivity or elevated levels of metals or salts. Generally, the toxicity classification of the leachates implies that although filter treatment with pine bark removes metal contaminants from the leachates effectively, it does not alter leachate toxicity noticeably. The leachates with the highest conductivity, pH and metal concentrations are most strongly correlated with an increased toxic response in the score plots of both untreated and treated leachates. This is in line with the toxicity classification of the leachate samples. The results from this study highlight the importance of evaluating treatment efficiency from the perspective of potential recipient effects, rather than in terms of residual concentrations of individual contaminants when treating waters with a complex contamination matrix, such as landfill leachates.

Interactions between pH, potassium, calcium, bromide, and phenol and their effects on the bioluminescence of Vibrio fischeri.

Little attention has been paid to how the light produced by the bacterium Vibrio fischeri in the Microtox assay is dependent on the concentration of essential ions such as sodium and potassium, and whether the concentrations of these ions affect the sensitivity of the test system to toxic chemicals. Five selected factors, pH, potassium (K(+)), calcium (Ca(2+)), bromide (Br(-)), and phenol (Phe), were simultaneously varied over a set of systematically planned experiments according to a D-optimal design that supported the estimation of a model with linear, quadratic, and two-factor interatcions of the studied factors. The bacterial light production represented by the gamma values in the Microtox assay for the 24 selected combinations of factors was measured at 5 and 15 min. The gamma values varied from negative to positive values greater than 1, indicating stimulation and inhibition of bacterial light production, respectively. The relationship between the gamma values and the factor settings was investigated with multiple linear regression. After 5 min of exposure, the light production was significantly affected by linear and quadratic terms for K(+), pH, and Phe and an interaction between pH and Phe. The situation was more complex after 15 min of exposure, since in addition significant interactions were found for K x Phe and Ca x pH. The tolerance of V. fischeri to Phe was enhanced by increasing the K and Ca concentrations. Data indicate that the ion composition and pH of the sample, as well as the diluents, need to be considered when the toxicity of salts, water samples, and extracts of sediments and soils are tested using commercially certified toxicity test kits.

Environmental hazard screening of a metal-polluted site using pressurized liquid extraction and two in vitro bioassays.

Rapid screening methods can improve the cost effectiveness, throughput, and quality of risk assessments of contaminated sites. In the present case study, the objective was to evaluate a combination of pressurized liquid extraction and 2 in vitro bioassays for the hazard assessment of surface soil sampled from 46 points across a pyrotechnical industrial site. Pressurized liquid extraction was used to rapidly produce soil-water extracts compatible with 2 high-capacity bioassays. Hazard assessment using combined toxicological and chemical screening revealed zones with relatively high potential risks of metal pollution. Multivariate data analysis provided indications that significant inhibition in the bioassays was correlated with levels of metals in the extracts, suggesting an elevated toxic potential from certain metals. Low pH and high concentrations of dissolved organic carbon were associated with increased cytotoxicity of extracts, indicating that these factors influence metal bioavailability. The cytotoxicity observed was more strongly correlated to metal concentrations in the extracts than in the soil, suggesting that measurements of total metal concentrations in soils do not provide good indications of the soil's potential toxicity.

Improving soil investigations at brownfield sites using a flexible work strategy and screening methods inspired by the US Environmental Protection Agency's triad approach.

Investigations of polluted brownfield sites and sample analyses are expensive, and the resulting data are often of poor quality. Efforts are needed, therefore, to improve the methods used in investigations of brownfield sites to both reduce costs and improve the quality of the results. One approach that could be useful for both of these purposes is the triad strategy, developed by the US Environmental Protection Agency, in which managing uncertainty is a central feature. In the investigations reported here, a field study was conducted to identify possible ways in which uncertainties could be managed in practice. One example considered involves optimizing the uncertainty by adjusting the sizes of samples and the efforts expended in analytical work according to the specific aims of the project. In addition, the potential utility of several toxicity assessment methods for screening sites was evaluated. As well as presenting the results of these assessments, in this contribution we discuss ways in which a flexible work strategy and screening methods inspired of the triad philosophy could be incorporated into the Swedish approach to remediate brownfield sites. A tiered approach taking advantage of field and screening methods is proposed to assess brownfield sites focusing on the response and acceptable uncertainty that are required for the task.

Potential toxic effect on aquatic fauna by the dwarf shrub Empetrum hermaphroditum.

The common evergreen dwarf shrub Empetrum hermaphroditum has influence on the functioning of boreal terrestrial ecosystems in northern Sweden. The negative effects of E. hermaphroditum are partly attributed to the production of the dihydrostilbene, batatasin-III, which is released from leaves and litter by rain and snowmelt. In this study, we investigated whether batatasin-III is carried by runoff into streams and lakes during the snowmelt period and whether it is also potentially hazardous to aquatic fauna. Sampling of water from streams and a lake for which the surrounding terrestrial vegetation is dominated by E. hermaphroditum was done during the snowmelt period in May 1993 and in 1998, and analyzed for batatasin-III. Using 24- and 48-hr standard toxicity tests, we analyzed toxicity to brown trout (Salmo trutta) alevins and juvenile water fleas (Daphnia magna). Toxicity (proportion of dead individuals) to trout was tested at pH 6.5 and compared with that of a phenol within a range of concentrations. In the toxicity (proportion of immobilized individuals) test on D. magna, the interactive effect of pH (pH 5.5-7.0) was included. Concentration of batatasin-III was generally higher in 1998 than in 1993 and showed peak levels during snowmelt. Concentration in ephemeral runnels > the lake > streams running through clear-cuts dominated by E. hermaphroditum > control streams lacking adjacent E. hermaphroditum vegetation. The maximum concentration of batatasin-III found was 1.06 mg l(-1). The proportion of dead yolk sac alevins increased significantly (P < 0.001) with increasing concentrations of batatasin-III and time of exposure. After 24 hr, EC50 was 10 mg l(-1). It was 2 mg l(-1) after 48 hr. The effect of phenol was negligible, indicating a specific phytotoxic effect of the bibenzyl structure of batatasin-III. The proportion of mobile D. magna became significantly smaller (P < 0.001) with increasing concentrations of batatasin-III, with decreasing pH, and with increasing exposure time. EC50 varied between 7 and 17 mg l(-1) at pH 5.5 and 7.0, respectively. After 24 hr EC50 decreased and was 2.5 at pH 5.5 and 12 mg l(-1) at pH 7.0. The levels of batatasin-III found in the field samples were below the lowest EC50 in acute toxicity tests. However, in view of the interactive effect of pH and exposure time, this study suggests that this stable plant metabolite may impose a lethal effect on the aquatic fauna in small streams.