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. 相似文献
In this chapter, the concept of exposure assessment and its evolution is introduced, and evaluated by critically appraising the pertinent literature as it applies to exposures to Particulate Matter (PM). Exposure measurement or estimation methodologies and models are reviewed. Three exposure/measurement methodologies are assessed. Estimation methods focus on source evaluation and attribution, sources include those outdoors and indoors as well as in occupational and in-transit environments. Fate and transport models and their inputs are addressed to estimate concentrations outdoors and indoors; source attribution techniques help focus on the contributing sources. Activity pattern techniques are also reviewed and their use in exposure models to estimate inhalation exposure to PM is presented. Deterministic, regression and other stochastic models of exposure to PM are reviewed and evaluated. Strengths, limitations, assumptions and affirmations of the use of exposure assessment as an integral component of risk assessment and risk management are discussed in the conclusions and discussions section of this work. 相似文献
Goal, Scope and Background In order to evaluate the estrogenic activity of sediments and XAD water extracts of selected sites of the catchment area
of the River Neckar, a river system in Southern Germany, an integrative assessment approach was used to assess the ecological
hazard potential of endocrine-disrupting compounds in sediment and water.
Methods The approach is based on estrogen receptor-mediated vitellogenin synthesis induced in isolated hepatocytes of rainbow trout
and quantified in a non-radioactive dot blot/RNAse protection-assay in parallel to comprehensive chemical analyses of estrogenic
substances.
Results and Discussion Numerous investigated extracts revealed an estrogen activity comparable to that of the positive control (1 nM 17?-estradiol
corresponding to 270 ng/L in the test medium). Based on a concentration factor of 30 in the extracts and a recovery of XAD
resins of approximately 80 %, 17?-estradiol equivalent concentrations between 20 and 26.7 ng/L could be calculated downstream
of a sewage treatment plant (< 0.1 ng/L for a reference site). A comparison of the bioassay-derived Bio-TEQs (toxicity equivalents)
and the Chem-TEQs revealed a high correlation with a Pearson coefficient of 0.85, indicating that the same ranking of the
samples could be obtained with respect to the endocrine disrupting potential with both chemical and bioanalytical analysis.
However, the TEQ concentrations computed from chemical analyses were significantly lower than the bioassay-derived TEQ concentrations.
In fact, in none of the samples, more than 14 % of the vitellogenin-inducing potency could be attributed to the substances
(steroids, alkylphenols, bisphenol A, diethylstilbestrol) analyzed.
A comparison of the endocrine disrupting potential of sediments extracted by the solvents acetone and methanol revealed lower
biological effects for acetone-extracted samples. Possible reasons may be a masking of endocrine effects in acetone extracts
by cytotoxicity, a low extraction efficiency of the solvent acetone, or anti-estrogen potencies of some extracted sediment
compounds.
Using a mass balance approach, the contribution of the compounds analyzed chemically (Chem-TEQs) to the total endocrine activity
(Bio-TEQs) was calculated. Based on the very low detection limits, particularly of the steroids with their high TEF factors,
results revealed that a calculation of the Chem-TEQs is associated with considerable scale inaccuracy: Whereas only 7-15 %
of the biological effectiveness (Bio-TEQs) could be explained by endocrine substances identified above the detection limits,
the assumption of concentrations slightly below the given detection limits would result in a significant over estimation (137-197
%) of the Bio-TEQs. Even the interassay variation of the dot blot assay with different fish donors for primary hepatocyte
(factor 2 - 2.5) is relatively low, when compared to the large range of the Chem-TEQ concentrations (factor 20) obtained when
applying different modes of calculation.
Conclusions and Outlook Overall, only a minor portion of the endocrine activity detected by bioassays could be linked to compounds identified by
chemical analysis. In vitro assays for assessment of endocrine activities are useful as sensitive integrating methods that
provide quantitative estimates of the total activity of particular receptor-mediated responses. Although discrepancies may
also result from different bioanalytical approaches, it is overall likely that bioanalytical and not chemical analytical approaches
give the correct estimate of endocrine disrupting potencies in environmental samples. As a conclusion, assessment of endocrine
disruption based on chemical analysis alone does not appear sufficient and further research into the spectrum of substances
with potential endocrine activity as well as into additive or even synergistic effects in complex environmental samples is
urgently needed. 相似文献
The fate of the explosive 2,4,6-TNT in plants is of major interest. Therefore, a method was developed to analyse TNT and derivatives in plant tissue. The method was utilized to investigate the uptake and metabolism of TNT inMedicago sativa andAllium schoenoprasum grown in hydroponic cultures containing TNT levels of 0.1 to 10 mg/1. Detectable concentrations of nitrotoluenes were significantly higher inAllium schoenoprasum than inMedicago sativa. The uptake of TNT in plants was directly related to the initial TNT level. The principal nitroaromatic components in roots and shoots of both plant species were identified as 4-ADNT and 2-ADNT in equal amounts, with substantially less TNT. 相似文献
Monitoring of immission of persistent organic pollutants in the industrialized area of Volta Redonda (V.R.) and in the National Park of Itatiaia (PNI) in southeast Brazil was performed using an endemic bromeliad species as biomonitor and measuring bulk deposition rates of polychlorinated biphenyls (PCB) and polycyclic aromatic hydrocarbons (PAH). For the sum of PCB, overall deposition rates were between 17 and 314 ng/(m2 day) in winter and between 43 and 81 ng/(m2 day) in summer, respectively. Deposition rates of dioxin-like PCBs ranged from 0.14 to 2.8 pg WHO-TEQ/(m2 day) in winter and from 0.90 to 4.3 pg WHO-TEQ/(m2 day) in summer. PCB deposition rates (total PCB and WHO-TEQ) were in the same range in winter in V.R. and PNI. In summer, contamination levels in V.R. were 6-10-folds higher than in PNI. PCB concentrations in biomonitor samples from V.R. and PNI were in the same range in summer and in winter. Concentrations of total PCB ranged from 14 to 95 microg/kg dry matter (d.m.) in winter and from 18 to 27 microg/kg d.m. in summer, respectively. The TEQ values were between 1.7 and 4.1 ng WHO-TEQ/kg d.m. in winter and between 1.9 and 2.9 ng WHO-TEQ/kg d.m. in summer. PCB concentrations of di-ortho PCB but not of non-ortho PCB were a factor of 2-4 lower in summer in both areas. PCB congener profiles resembled those from technical formulations. The profiles shifted to the higher chlorinated congeners in summer, probable due to revolatilisation of the lighter components at higher temperatures. PCB profiles in biomonitor resembled those from deposition samples and the shift to the heavier congeners in summer was even more pronounced. PAH deposition rates were in a similar range in both areas (131-2415 ng/(m2 day)). PAH levels in biomonitor samples from V.R. were about one order of magnitude higher than in samples from PNI indicating the impact of local sources. PAH profiles revealed stationary thermal processes as main source of contamination in V.R. whereas in PNI, biomass burning seems to be the main contamination source. 相似文献
A new analytical method using accelerated solvent extraction was developed for the determination of 10 particle-associated polar and semipolar pesticides. In addition, six deuterated analogues of the target compounds were evaluated as internal standards. The method yielded acceptable accuracy (73–103% recovery) and precision (<25% relative standard deviation) for eight compounds. Using size exclusion chromatography (SEC) as cleanup step resulted in higher recoveries compared to solid phase extraction (SPE) cleanup.
Deuterated standards with 10 or more deuterium atoms performed well as internal standards concerning similar recovery and correlation with the target analytes.
The method was employed to extract particle-associated pesticides from 16 streams located in an area with intense agriculture in France. Acetochlor, pirimicarb, tebuconazole, fenpropidin, -endosulfan and chlorfenvinphos were detected at concentrations up to 1 mg kg−1 dry weight. A comparison with aquatic toxicity data indicated potential risk to the benthic fauna exposed to these concentrations of pirimicarb, -endosulfan and chlorfenvinphos.
We suggest that the method presented here be used for the extraction and quantitation of particle-associated polar pesticides. 相似文献
In this study, metals (Be, Cr, Mn, Fe, Ni, Cu, Zn, Ag, Cd, Pb and Hg) in the fine-grained fraction (<63 microm) from 12 sites at different locations in northern San Francisco Bay over a year period from March 2000 to March 2001 were analyzed after acid extraction. The results showed that metal concentrations in the sediments varied from site to site, whereas some of them were found elevated with respect to the sediment of Tomales Bay, CA, which has little contamination history, indicating an enrichment of the metals in the sediment samples analyzed. Sediment toxicity and bioaccumulation evaluation by a clam species, Macoma nasuta, exposed to the sediment samples collected from the six sampling sites was carried out. The results showed that the sediment samples tested significantly reduced clam survival. Toxicity of the sediments to the clam was, in part, related to elevated metal concentrations in the sediments. In order to examine geochemistry of the metals and to understand potential correlations between metal concentrations and geochemical matrix elements of the sediments, bioavailability and toxicity of the metals, detailed analysis of metal concentrations associated with total organic carbon and the Fe-oxy-hydroxides in the sediment samples was performed. The analysis showed that sediment geochemistry appeared to influence metal bioavailability and may have important impacts on the toxicity of these metals to the clam. 相似文献
Surface water methane (CH4) and nitrous oxide (N2O) concentrations and fluxes were investigated in two subtropical coastal embayments (Bramble Bay and Deception Bay, which are part of the greater Moreton Bay, Australia). Measurements were done at 23 stations in seven campaigns covering different seasons during 2010-2012. Water-air fluxes were estimated using the Thin Boundary Layer approach with a combination of wind and currents-based models for the estimation of the gas transfer velocities. The two bays were strong sources of both CH4 and N2O with no significant differences in the degree of saturation of both gases between them during all measurement campaigns. Both CH4 and N2O concentrations had strong temporal but minimal spatial variability in both bays. During the seven seasons, CH4 varied between 500% and 4000% saturation while N2O varied between 128 and 255% in the two bays. Average seasonal CH4 fluxes for the two bays varied between 0.5 ± 0.2 and 6.0 ± 1.5 mg CH4/(m2·day) while N2O varied between 0.4 ± 0.1 and 1.6 ± 0.6 mg N2O/(m2·day). Weighted emissions (t CO2-e) were 63%-90% N2O dominated implying that a reduction in N2O inputs and/or nitrogen availability in the bays may significantly reduce the bays' greenhouse gas (GHG) budget. Emissions data for tropical and subtropical systems is still scarce. This work found subtropical bays to be significant aquatic sources of both CH4 and N2O and puts the estimated fluxes into the global context with measurements done from other climatic regions. 相似文献
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. 相似文献
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. 相似文献