Eija Schultz

Behavior of titanium dioxide nanoparticles in Lemna minor growth test conditions

Titanium dioxide nanoparticles (TiO(2) NPs) have raised concern of environmental risks due to their widespread applications, but little is known about the potential toxicity of TiO(2) NPs to aquatic plants. The aim of this work was to study the effects of TiO(2) NPs on Lemna minor and to study the behavior of TiO(2) NPs under modified ISO 20079 test conditions. TiO(2) NPs had a tendency to aggregate in ISO (Steinberg) growth medium, but modification of the standard growth medium enabled the exposure of L. minor to TiO(2) NPs. By dilution of the growth medium (1:10), and exposure under semi-static conditions with medium renewal every second or third day, the size of TiO(2) particles remained rather stable throughout the test period. TiO(2) NPs showed no adverse effect on the growth rate or chlorophyll a content of L. minor, even at a high exposure concentration of 5 mg L(-1) and extended exposure time of 14 days. TiO(2) NPs attached onto L. minor cell walls, but no cellular uptake was observed. Although TiO(2) NPs were not toxic to L. minor, the potential transfer of TiO(2) NPs in aquatic food chains, e.g. attached to the plant leaves and other biological surfaces may be of importance, causing exposure of other organisms and contributing to the environmental fate of nanoparticles.

Comparisons of terrestrial and aquatic bioassays for oil-contaminated soil toxicity

BackgroundPetroleum products are widely used in various sections of industry and they are one of the most abundant sources of environmental contamination. These products are classified by their physico-chemical properties such as boiling point, density and viscosity. Oil contamination in the environment is primarily evaluated by measuring the chemical concentrations of petroleum products in the solid or water phase. The results of chemical analyses do not correspond directly with the harmful environmental effects of petroleum products on the soil flora and fauna, because the interactions between oil compounds and the production of their methabolites in soil are not measured in chemical assessments. These kinds of effects of complex chemical mixtures in soil can be estimated by bioassays. Therefore, ecotoxicological tests are important for estimating soil quality in the risk assessment of oil-contaminated soil sites.ObjectivesThe objective of this study was to examine the oil-contaminated soil site of a closed petrol station with both chemical and ecotoxicological methods. The goals of this study were to compare the sensitivity of the terrestrial and aquatic bioassays and to compare the toxicity responses of aquatic bioassays determined from three different extraction procedures. In addition, our aim was to characterise a cost-effective battery of bioassays that could be applied to a comparison of oil-contaminated soils. It was in our interest to investigate oil-contaminated soil with oil concentrations of 2500–12000 mg/kg and to find out the possible differences between terrestrial and aquatic toxicity tests.MethodsSix soil samples from a closed petrol station were examined for toxicity with terrestrial and aquatic tests. Terrestrial tests includedEnchytraeus albidus survival and reproduction assays and seed germination assays using wheat, cress, lettuce, and red clover seeds and growth inhibition assays of onions. The toxicities of the water-extractable fractions of the soil samples obtained from three different extractions were tested with aquatic bioassays based on plants (onion and duckweed growth inhibition tests), microbes (luminescent bacteria test), and enzyme inhibition (reverse electron transport test, RET). Chemical analyses of the solid samples were carried out simultaneously. Results. Oil concentrations ranged from 2500 to 12000 mg/kg, BTEX varied from 300 to 2800 mg/kg, and fuel additives: MTBE and TAME from 0.0 1 to 260 mg/kg. Only the sample contain-ing 12000 mg/kg oil had a significant toxic impact on all test organisms. Soil samples with oil concentrations 2500–6200 mg/ kg had no or only slight adverse effects on the test organisms with one exception, theE. albidus reproduction test. TheE. albidus survival and reproduction tests were the most sensitive bioassays of the terrestrial tests, and the luminescent bacteria test of the aquatic tests.

Estrogenic activity in Finnish municipal wastewater effluents.

Effluents from wastewater treatment plants (WWTPs) are a major source of estrogenic compounds to the aquatic environment. In the present work, estrogenic activities of effluents from eight municipal WWTPs in Finland were studied. The main objectives of the study were to quantify the concentrations of selected estrogenic compounds, to evaluate their contribution to estrogenic potency and to test the feasibility of the commercial bioassays for wastewater analysis. The effluent samples were analyzed by two in vitro tests, i.e. ERα-CALUX(®) and ELISA-E2, and by liquid chromatography mass spectrometry for six estrogenic compounds: estrone (E1), 17β-estradiol (E2), estriol (E3), 17α-ethinylestradiol (EE2), 17α-estradiol and bisphenol A (BPA). Estrogenic effects were found in all of the effluent samples with both of the bioassays. The concentrations measured with ELISA-E2 (8.6-61.6 ng/L) were clearly higher but exhibited a similar pattern than those with chemical analysis (E2 <limit of quantification - 6.8 ng/L) and ERα-CALUX(®) (0.8-29.7 ng E2 EEQ/L). Due to the concentrations under limit of quantification, the evaluation of the chemical contribution to estrogenic potency was possible only for E1 and BPA, which contributed less than 10% to the observed effects, except in one sample with a high BPA contribution (17%). The contribution of E2 was significant in two samples where it was detected (28% and 67%). The results demonstrated that more comprehensive information on potential estrogenic activity of wastewater effluents can be achieved by using in vitro biotests in addition to chemical analysis and their use would be beneficial in monitoring and screening purposes.

Composting of forest industry waste water sludges for agricultural use

Abstract The feasibility of composting of pulp and paper industry sludges was studied. The aim was to study the progress of the composting process and the toxicity and applicability of the sludges in agriculture. Five different sludges were used: two biosludges and three biosludge and primary sludge mixtures. The sludges were composted in seven piles, two smaller of them founded with leachates collection systems. The piles were monitored throughout the composting period by physical measurements, chemical analyses and toxicity tests. Also the lignin concentrations were measured from the sludges and from the soil as well as the amount of mycelium. The results showed that all sludges were easily composted and they will be suitable for the use as soil improvement in agricultural plant production. Toxicity of the sludges decreased during composting and all the sludges were proved to be non-toxic at the end of the composting. Heavy metal concentrations were low in all sludges and leachates. The leachates were also non-toxic and the concentrations of nutrients were low.

Industrial waste leachates: toxicity detection with microbiotests and biochemical tests

This study deals with the toxicity of the leachates of different solid industrial wastes. The leachates represent 4 types of industrial waste: oily waste resulting from machining of metal blades, fly ash from the power plant of a mill producing viscose fibres, de-inking waste from a secondary pulp-production process, and jarosite, a byproduct from a nickel and cobalt mill. Several bioassays were performed on the leachates: the submitochondrial reverse electron transfer test (RET), the Lemna minor growth inhibition test, seed germination tests and 4 commercial microbiotests (MetPLATE, MetPAD, ToxiChromotest and Mutatox). The leachate samples differed in their toxicity profiles, thus confirming the differences observed in their chemical composition. In decreasing order, the toxicity of the wastes was as follows: jarosite, fly ash, oily waste from metal blades and de-inking-waste.

Assessment of Ecological Risks at Former Landfill Site Using TRIAD Procedure and Multicriteria Analysis

Old industrial landfills are important sources of environmental contamination in Europe, including Finland. In this study, we demonstrated the combination of TRIAD procedure, multicriteria decision analysis (MCDA), and statistical Monte Carlo analysis for assessing the risks to terrestrial biota in a former landfill site contaminated by petroleum hydrocarbons (PHCs) and metals. First, we generated hazard quotients by dividing the concentrations of metals and PHCs in soil by the corresponding risk-based ecological benchmarks. Then we conducted ecotoxicity tests using five plant species, earthworms, and potworms, and determined the abundance and diversity of soil invertebrates from additional samples. We aggregated the results in accordance to the methods used in the TRIAD procedure, conducted rating of the assessment methods based on their performance in terms of specific criteria, and weighted the criteria using two alternative weighting techniques to produce performance scores for each method. We faced problems in using the TRIAD procedure, for example, the results from the animal counts had to be excluded from the calculation of integrated risk estimates (IREs) because our reference soil sample showed the lowest biodiversity and abundance of soil animals. In addition, hormesis hampered the use of the results from the ecotoxicity tests. The final probabilistic IREs imply significant risks at all sampling locations. Although linking MCDA with TRIAD provided a useful means to study and consider the performance of the alternative methods in predicting ecological risks, some uncertainties involved still remained outside the quantitative analysis.

Evaluation of biodegradation of nonylphenol ethoxylate and lignin by combining toxicity assessment and chemical characterization

The aerobic biodegradation of commercial nonylphenol ethoxylate (NPE) mixture and alkali lignin was studied using the OECD headspace test accompanied by the simultaneous measurement of ecotoxicity directly from the biodegradation liquors and by the follow-up of the chemical composition of the studied chemicals. NPE degradation was dependent on the inoculum source: approximately 40% of NPE was mineralized into CO(2) during the 4-week experiment when inoculum from Helsinki City wastewater treatment plant (WWTP) was used, and only 12% was mineralized when inoculum from Jyväskylä City WWTP was used. Chemical analyses revealed a shift in the ethoxylate chain length from longer to shorter soon after the beginning of the NPE biodegradation tests. At the same time also toxicity (reverse electron transport assay, RET) and estrogenic activity (human estrogen receptor yeast) measured directly from the biodegradation liquors decreased. In case of alkali lignin, approximately 11% was mineralized in the test and chemical analysis showed in maximum a 30% decrease in lignin concentration. Toxicity of lignin biodegradation liquors started to decrease in the beginning of the test, but became more toxic towards the end of the test again. Especially RET assay proved to be sensitive enough for measuring toxicity changes directly from biodegradation liquors, although a concentrating treatment of the liquors is recommended for a more detailed characterization and identification of toxic metabolites.

Biological effects of high molecular weight lignin derivatives.

A number of high molecular weight (HMW) lignin derivatives possessing varied chemical properties were screened for their biological effects in order to obtain more information on the possible structural features of HMW lignin-related effects. The studied compounds were both commercial and in-house extracted lignin derivatives. Bioassays used include reverse electron transport (RET), Vibrio fischeri, Daphnia magna, and juvenile rainbow trout (Oncorhynchus mykiss) hepatocytes. The studied lignin derivatives inhibited the in vitro systems and luminescence of V. fischeri bacteria to some extent-daphnids were not affected. It seems that, at least in the RET assay, certain pH-dependent functional groups in lignin may be of importance regarding the biological effects.

More accuracy to the EROD measurements--the resorufin fluorescence differs between species and individuals.

Ethoxyresorufin-O-deethylase (EROD) activity is a biomarker of exposure to planar aromatic hydrocarbons, and it is often measured from the S9 fraction. The effect of the liver S9 fraction of seven boreal freshwater fish species on the fluorescence of resorufin was studied. The S9 fractions diminished resorufin fluorescence by 40-80%, and there were large differences between species. Thus, using a resorufin standard curve without the S9 fraction leads to a large underestimation of the EROD activity. Therefore a microwell plate EROD method was developed that takes into account the effect of each sample on resorufin fluorescence. At least two mechanisms were involved in the decrease of the fluorescence: opaqueness of the sample, and enzymes (DT-diaphorase and plausibly NADPH-CYP450 oxidoreductase) that reduce resorufin to a non-fluorescent form.

Weight-of-evidence approach in assessment of ecotoxicological risks of acid sulphate soils in the Baltic Sea river estuaries.

Acidity and leaching of metals from acid sulphate soils (ASSs) impair the water quality of receiving surface waters. The largest ASS areas in Europe are found in the coasts of the northern Baltic Sea. We used weight-of-evidence (WoE) approach to assess potential risks in 14 estuary sites affected by ASS in the Gulf of Finland, northern Baltic Sea. The assessment was based on exposure and effect profiles utilizing sediment and water metal concentrations and concurrent pH variation, sediment toxicity tests using the luminescent bacterium Vibrio fischeri and the midge Chironomus riparius, and the ecological status of benthic macroinvertebrate communities. Sediment metal concentrations were compared to national sediment quality criteria/guidelines, and water metal concentrations to environmental quality standards (EQSs). Hazard quotients (HQs) were established for maximum aluminium, cadmium and zinc concentrations at low pH based on applicable US EPA toxicity database. Sediment metal concentrations were clearly elevated in most of the studied estuaries. The EQS of cadmium (0.1 μg/l) was exceeded in 3 estuaries out of 14. The pH-minima were below the national threshold value (5.5) between good and satisfactory water quality in 10 estuaries. V. fischeri bioluminescence indicated toxicity of the sediments but toxic response was not observed in the C. riparius emergence test. Benthic invertebrate communities were deteriorated in 6 out of 14 sites based on the benthic invertebrate quality index. The overall ecotoxicological risk was assessed as low in five, moderate in three and high in five of the estuary sites. The risk assessment utilizing the WoE approach indicated that harmful effects of ASSs are likely to occur in the Baltic Sea river estuaries located at the ASS hotspot area.