Integral assessment of estrogenic potentials in sediment-associated samples

Background, aim, and scope  The enzyme-linked receptor assay (ELRA) detects estrogenic and anti-estrogenic effects at the molecular level of receptor binding and is a useful tool for the integrative assessment of ecotoxicological potentials caused by hormonally active agents (HAA) and endocrine disrupting compounds (EDC). The main advantage of the ELRA is its high sample throughput and its robustness against cytotoxicity and microbial contamination. After a methodological adaptation to salinity of the ELRA, according to the first part of this study, which increased its salinity tolerance and sensitivity for 17-β-estradiol, the optimised ELRA was used to investigate 13 native sediments characterised by different levels of salinity and chemical contamination. The applicability of the ELRA for routine analysis in environmental assessment was evaluated. Salinity is often a critical factor for bioassays in ecotoxicological sediment assessment. Therefore, salinity of the samples was additionally adjusted to different levels to characterise its influence on elution and binding processes of receptor-binding substances. Materials and methods  The ELRA was carried out with the human estrogen receptor α (ER) in a 96-well microplate format using the experimental setup known from the competitive immunoassay based on ligand–protein interaction. It is an important improvement that a physiologically relevant receptor was used as a linking protein instead of an antibody. The microplates were coated with a 17-β-estradiol-BSA conjugate, and dilution series of estradiol and of native sediment samples were added and incubated with the ER. After a washing step, a biotinylated mouse anti-ER antibody was added to each well. Receptor binding to estradiol, agonistic and antagonistic receptor binding, were determined by a streptavidin-POD-biotin complex with subsequent measurement of the peroxidase activity at the wavelength of 450 nm using a commercial ELISA multiplate reader. The sediment elutriates and pore water samples of sediments were tested in a dilution series to evaluate at which dilution step the receptor-binding potential ends. In the elution process (see Section 2.1 to 2.2), a method was developed to adjust the salinity to the levels of the reference testings, which offers an appropriate option to adjust the salinity in both directions. Statistical evaluation was made with a combination of the Mann–Whitney U test and the pT-method. Results  This part of the study characterised the environmental factor ‘salinity’ for prospective applications of the ELRA. Using reference substances such as 17-β-estradiol, the ELRA showed sigmoid concentration-effect relations over a broad range from 0.05 μg/l to 100 μg/l under physiological conditions. After methodological optimisation, both sensitivity and tolerance of the assay against salinity could be significantly raised, and the ELRA became applicable under salinity conditions up to concentrations of 20.5‰. The mean relative inter-test error (n = 3) was around 11% with reference substances and below 5% for single sediments elutriates in three replicates each. For sediment testings, the pore water and different salinity-adjusted elutriates of 13 sediments were used. A clear differentiation of the receptor-binding potential could be reached by application of the pT-method. Thereby, pT-values from one to six could be assigned to the sediments, and the deviation caused by the different salinity conditions was one pT-value. The mean standard deviation in the salinity adaptation procedure of the elutriates was below 5%. Discussion  Although the ELRA has already been used for assessments of wastewater, sludge and soil, its applicability for samples to different salinity levels has not been investigated so far. Even if the ELRA is not as sensitive as the E-screen or the YES-assay, with regard to reference substances like 17-β-estradiol, it is a very useful tool for pre-screening, because it is able to integrate both estrogenic as well as anti-estrogenic receptor-binding effects. According to the results of sediment testing, and given the integrative power to detect different directions of effects, the ELRA shows sufficient sensitivity and salinity tolerance to discriminate receptor-binding potentials in environmental samples. Conclusions  The optimised ELRA assay is a fast, cost-effective, reliable and highly reproducible tool that can be used for high-throughput screening in a microplate format in detecting both estrogenic and anti-estrogenic effects. Additionally, the ELRA is robust against microbial contaminations, and is not susceptible towards cytotoxic interferences like the common cell-culture methods. The general applicability and sufficient sensitivity of the ELRA was shown in freshwater environments. Marine and brackish samples can be measured up to salinity levels of 20.5‰. Recommendations and perspectives  In view of the proven sensitivity, functionality and the fastness of the ELRA, it is recommendable to standardise the test method. At the moment, no adequate in vitro test procedure exists which is standardised to DIN or ISO levels. The E-screen and the yeast estrogen/androgen screens (YES/YAS) sometimes underlie strong cytotoxic effects, as reported in the first part of this study. Further development of an ELRA assay using human androgen receptors appears to be very promising to gain information about androgenic and anti-androgenic effects, too. This would offer a possibility to use the ELRA as a fast and reliable pre-screening tool for the detection of endocrine potentials, thus minimising time and cost-expensive animal experiments.     

Activated carbon amendment to sequester PAHs in contaminated soil: a lysimeter field trial

Activated carbon (AC) amendment is an innovative method for the in situ remediation of contaminated soils. A field-scale AC amendment of either 2% powder or granular AC (PAC and GAC) to a PAH contaminated soil was carried out in Norway. The PAH concentration in drainage water from the field plot was measured with a direct solvent extraction and by deploying polyoxymethylene (POM) passive samplers. In addition, POM samplers were dug directly in the AC amended and unamended soil in order to monitor the reduction in free aqueous PAH concentrations in the soil pore water. The total PAH concentration in the drainage water, measured by direct solvent extraction of the water, was reduced by 14% for the PAC amendment and by 59% for GAC, 12 months after amendment. Measurements carried out with POM showed a reduction of 93% for PAC and 56% for GAC. The free aqueous PAH concentration in soil pore water was reduced 93% and 76%, 17 and 28 months after PAC amendment, compared to 84% and 69% for GAC. PAC, in contrast to GAC, was more effective for reducing freely dissolved concentrations than total dissolved ones. This could tentatively be explained by leaching of microscopic AC particles from PAC. Secondary chemical effects of the AC amendment were monitored by considering concentration changes in dissolved organic carbon (DOC) and nutrients. DOC was bound by AC, while the concentrations of nutrients (NO(3), NO(2), NH(4), PO(4), P-total, K, Ca and Mg) were variable and likely affected by external environmental factors.     

Passive dosing: an approach to control mutagen exposure in the Ames fluctuation test

By using dialysis equilibrium experiments, the sorption of a branched nonylphenol isomer [4-(1-ethyl-1,3-dimethylpentyl)-phenol] (NP111) on various humic acids (HAs) isolated from river sediments and two reference HAs was studied. The HAs were characterized by solid-state ¹³C direct polarization/magic angle spinning nuclear magnetic resonance (¹³C DP/MAS NMR) spectroscopy. Sorption isotherms of NP111 on HAs were described by a linear model. The organic carbon-normalized sorption coefficient (K_OC) ranged from 2.3 × 10³ to 1.5 × 10⁴ L kg⁻¹. Interestingly, a clear correlation between K_OC value and alkyl C content was observed, indicating that the aliphaticity of HAs markedly dominates the sorption of NP111. These new mechanistic insights about the NP111 sorption indicate that the fate of nonylphenols in soil or sediment depends not only on the content of HA, but also on its structural composition.     

Biotoxizit?tstests mit PAK Die Nodulation in der Leguminosen-Rhizobien-Symbiose als sensibler Wirkungsparameter

Im Zusammenhang mit der Umweltchemikalienprüfung und der Untersuchung und Kontrolle kontaminierter B?den besteht gro?er Bedarf an Testverfahren mit h?heren Pflanzen, die schnell, einfach und preisgünstig durchzuführen sind und bereits bestehende Testverfahren nach OECD- und BBA-Richtlinien sinnvoll erg?nzen k?nnen. Dieser Beitrag stellt einen Biotoxizit?tstest vor, der die Nodulation in der Leguminosen-Kn?llchen-Symbiose als sensiblen Wirkungsparameter für die Belastung mit polyzyklischen aromatischen Kohlenwasserstoffen (PAK) benutzt. Fremdstoff, Pflanze und Mikroorganismus werden dabei auf einer festen Matrix in Kontakt miteinander gebracht; das axenische System erlaubt die Untersuchung der Fremdstoffwirkung sowohl auf den Gesamtorganismus Pflanze, als auch auf die Interaktion von Pflanze und Mikroorganismus (Kn?llchenbildung). In Dose-Response-Versuchen zeigte sich der Wirkungsparameter Nodulation, gemessen als Anzahl Wurzelkn?llchen pro Pflanze, sensibler als andere pflanzliche Wachstumsparameter. There is great need for toxicity tests with higher plants for the investigation and monitoring of chemicals in the environment — especially for tests which are quick, easy, and inexpensive. Such tests should add to and improve current guidelines, e.g., OECD- and BBA-Guidelines. This article describes the nodulation of leguminous plants withRhizobium bacteria used in a bioassay with polycyclic aromatic hydrocarbons (PAH). Plants growing on a solid medium were exposed simultaneously to bacteria and PAH. This axenic culture system allows to observe the impact of PAH on plant growth and the interaction between plant and bacteria (nodulation). Dose-response-experiments indicated that nodulation measured as number of nodules per plant, was more sensitive to the inhibition by PAH than other plant growth parameters.     

Quantification of nitrate leaching from German forest ecosystems by use of a process oriented biogeochemical model

Simulations with the process oriented Forest-DNDC model showed reasonable to good agreement with observations of soil water contents of different soil layers, annual amounts of seepage water and approximated rates of nitrate leaching at 79 sites across Germany. Following site evaluation, Forest-DNDC was coupled to a GIS to assess nitrate leaching from German forest ecosystems for the year 2000. At national scale leaching rates varied in a range of 0–>80 kg NO₃–N ha⁻¹ yr⁻¹ (mean 5.5 kg NO₃–N ha⁻¹ yr⁻¹). A comparison of regional simulations with the results of a nitrate inventory study for Bavaria showed that measured and simulated percentages for different nitrate leaching classes (0–5 kg N ha⁻¹ yr⁻¹:66% vs. 74%, 5–15 kg N ha⁻¹ yr⁻¹:20% vs. 20%, >15 kg N ha⁻¹ yr⁻¹:14% vs. 6%) were in good agreement. Mean nitrate concentrations in seepage water ranged between 0 and 23 mg NO₃–N l⁻¹.     

Resilience to natural hazards: How useful is this concept?

Resilience is widely seen as a desirable system property in environmental management. This paper explores the concept of resilience to natural hazards, using weather-related hazards in coastal megacities as an example. The paper draws on the wide literature on megacities, coastal hazards, hazard risk reduction strategies, and resilience within environmental management. Some analysts define resilience as a system attribute, whilst others use it as an umbrella concept for a range of system attributes deemed desirable. These umbrella concepts have not been made operational to support planning or management. It is recommended that resilience only be used in a restricted sense to describe specific system attributes concerning (i) the amount of disturbance a system can absorb and still remain within the same state or domain of attraction and (ii) the degree to which the system is capable of self-organisation. The concept of adaptive capacity, which has emerged in the context of climate change, can then be adopted as the umbrella concept, where resilience will be one factor influencing adaptive capacity. This improvement to conceptual clarity would foster much-needed communication between the natural hazards and the climate change communities and, more importantly, offers greater potential in application, especially when attempting to move away from disaster recovery to hazard prediction, disaster prevention, and preparedness.     

Is there a Relation Between Ecological Practices and Spirituality? The Case of Benedictine Monasteries

For decades there has been a controversial debate over how far religious faith communities are specifically engaged in ecological practices (EP). Therefore we studied four Austrian and two German Benedictine monasteries religious ethics and spirituality as a means of a driving force for initiating EP. We draw upon theories of organizational learning processes and capacity-building of sustainability to interpret our empirical findings. The majority of monasteries are highly engaged in EP, initiated either as an outcome of individual activities or through a specific mostly informally acting group, but rarely an organizationally or systematically integrated goal of the monasteries, or a focus of capacity building. Monasteries follow a technical and economic decision-making process in implementing EP. Spirituality plays a limited role in the initiation of EP. The environment of monasteries–acceptance or critique against ecological practices–influences the monasteries decisions. Institutionalization of ecological practices into the monasteries organizational structure is rare.