Bakterielle Biotestverfahren zur Bestimmung der Toxizität von Abfällen

For a risk assessment of wastes, fast and sensitive screening methods are required to detect possible toxic effects. In this study, different leachability methods (DIN 38414, part 4; EPA 1310; EPA 1320) were used to prepare aqueous leachates from different wastes; these leachates were rested in different bacterial toxicity test systems for possible toxic effects. As bacterial toxicity tests, the luminescent bacteria test and the growth inhibition test with activated sludge bacteria were used. The test systems showed a good agreement of the results from the different leaching methods, thereby indicating the reliability of the test systems used. The luminescent bacteria inhibition test showed a higher sensitivity than the growth inhibition test which is in good accordance with literature data. We conclude that both test systems are well suited to assess the toxic potential of aqueous waste leachates.     

Toxizitätsänderungen in Photoreaktionen von Azaarenen

The azaarenes and the structurally analogous PAH accompany each other and occur ubiquitously in nature. Photochemical conversions like direct photolysis or the photoreaction with nitrate are important natural degradation reactions of azaarenes. Furthermore, photoreactions with chlorine and bromine radicals can be expected in nature. The reactions generate a large number of oxidized, nitrated and halogenated products with unknown toxicity. In this paper, a first screening of changes is reported in acute toxicity during the course of the photochemical reactions. As test system luminescent bacteria are used. For quinoline, isoquinoline and phthalazine the investigations show different changes of the toxicity of the reaction mixtures in dependence on the respective reaction type. The toxicity of the reaction mixtures of quinazoline and quinoxaline increases in all reactions investigated. Furthermore the reactions of bromine radicals with azaarenes also show an increase of the toxicity in all cases. Compared with the azaarenes in all reactions products with significantly higher toxicity are formed.     

Neue Bewertung bei den Toxizitätsäquivalenten für Dioxine/Furane und für PCB durch die WHO

While the large amount of data available makes it possible to provide a statement concerning the effects of PCDD/PCDF on the WHO TEF-reappraisal regarding the observance of dioxin emission values from waste incineration, similar values for PCBs could not be determined accurately or could only be estimated roughly for the WHO, because only limited measurement values are available for these substances. Thanks to the present availability of such measurement values from the MVA in Bielefeld-Herford, Germany and the MVR in Hamburg, Germany, it is now possible to determine a direct relationship between the low PCDD/PCDF values and the 12 different PCBs which are taken into consideration by the WHO. As the results of these investigations have shown, the statement in Section 2 claiming that there are generally only very low levels of PCDD/PCDF emissions stemming from waste incineration plants, and that the 12 additional PCBs considered by the WHO as well as the WHO TEFs lead to no relevant increase in the evaluation/calculation of the actual emission values nor to a value above the dioxin (I-TEQ) threshold value as established in the 17th BImSchV (Germany Federal regulations responsible for protecting the population from emissions), can be confirmed. A dominant role of the WHO PCB-TEQs is seen to be played by the PCB-126. The exhaust gas measurements in waste incineration plants might therefore already be sufficient if merely the dioxin-like PCB-126 values were to be used in the calculation of the dioxin I-TEQ.     

Existierende dioxininventare weltweit und neue methodik zur erstellung von vergleichbaren und vollständigen emissionsinventaren

Presently, there are less than 20 inventories to estimate releases of dioxins and furans. According to a recent survey, 12,900 g TEQ of these unwanted byproducts are emitted into the atmosphere by 16 countries. Highest emissions are from the densely populated industrialized countries of the Northern hemisphere — Japan and the United States of America. Whereas the metal producing and recycling industry is the sector with the highest dioxin emissions in Europe, waste incineration is considered to be the major source in many other countries. Measures to reduce dioxin emissions have resulted in strong downward trends, as shown e.g. in Germany and Japan; however, potentials for further reduction have been identified. So far, countries have utilized own methods to calculate their dioxin emissions and the majority has addressed releases to air only. The future Stockholm Convention on POPs will require to continuously reduce dioxin emissions. In order to assist countries in inventory making, UNEP has produced the Toolkit, a methodology to establish comparable dioxin inventories that address releases to air, water and land, with products and in residues.     

Bestimmung von Phthalsäureestern in Lebensmitteln, Frauenmilch, Hausstaub und Textilien

25 food samples, 5 mother’s milk specimens, 4 dust samples, and 16 textiles were analysed for phthalic acid esters. Phthalic acid esters were detected in all samples, with di (2-ethylhexyl) phthalate (DEHP) and di-n-butyl phthalate (DBP) being the most abundant phthalates. Raw milk samples revealed average concentrations of total phthalate of 0.1 mg/kg. Retail milk did not contain higher loads than raw milk, even storing samples until their “best-before” date did not result in elevated levels. Skimmed milk was less contaminated than whole milk. The higher concentrations of DEHP and total phthalates in cream samples are due to their higher fat content. With concentrations up to 1.54 mg/kg, ground hazelnuts, almonds, and nutmeg in plastic packagings showed relatively high levels. In infant food, only traces of DEHP and DBP could be found while other phthalic acid esters were not detectable. The mother’s milk samples also exhibited only low amounts of approx. 0.1 mg/kg, thus indicating that there is no accumulation of phthalate esters in the human body. Extraordinarily high concentrations were found in dust samples; with the levels ranging from 300 to 5370 mg/kg and DEHP being the major compound. This leads to the conclusion that the air path must play a considerable role in the transfer of phthalic acid esters. As dust contains considerable amounts of textile fibres, textiles were also included in the investigation. The phthalic ester levels in the textile samples ranged from 3.42 to 34.44 mg/kg. Therefore, the high phthalate contaminations of dust cannot be explained by textile fibres.     

Analyse „stetigkeitsbasierter“ Methoden zur Bestimmung von Stofffrachten in Fließgewässern

Aim and background

Increasing ecological awareness puts the load as a tool for water quality assessment in the foreground. Therefore it is very desirable to find optimal procedures for any given situation on basis of existing data. Criteria will be derived to fill that desire.

Method

The load for a water ingredient is an integrated entity of time-continuous loads (product of concentration and flow) for a given time, i.?e. a year (yearly load). Normally you can only observe a sample (in a statistical meaning) of these data. Therefore, load calculations vary with the concrete sample given. Each such calculation represents an estimation of the load, afflicted with some uncertainty. This uncertainty can be quantified, at least approximatively, by adding variance (scatter around “expected” mean) and the square of bias (divergence of “expected” mean from true mean). This quantity is called “mean squared error”. The methodological results will be illustrated by examples.

Focus

The main focus lies on the methodologies, which, besides data of concentrations observed discretely in time, additionally use quasi-continuously recorded data of other variates, namely the flow. For that situation, statistical models are developed and the load derived and estimated according to existing data. Hereby it is seen that all model based estimation formulae can be described as standard sampling formulae plus some additional terms which stem from comparison of some measurements (i.?e. mean) for quasi-continuous and discrete samples. This way a “continuity-correction” is performed on the standard formulae for the sampling case. This method is called “continuity-based” load determination.

Results

A surprising result is that no complicated mathematical-statistical models are necessary to determine the load. A simple linear concentration- or load-flow-model is sufficient. Additional variates lead, most of the time, to a larger mean squared error in load estimation, even when it is well known that they cannot detoriate the model fit. Only models which describe the concentration extraordinarily precise, near perfection, result in a smaller error.

Implications

It is enough to use one of the given formulae with some “continuity correction” to calculate a load. That can be done without using any software.     

Screening zur Mobilisierbarkeit von Kupfer aus Böden und Schlämmen

In the presented paper, the pHstat-procedure is compared with four simple extraction procedures. These procedures make use of the following extraction solutions: distilled water (based on DIN 38414 part 4, DIN-S4), 1 mol/L ammonium nitrate, 1 mol/L ammonium acetate at pH 7.0 and 1 mol/L sodium acetate at pH 5.0, respectively. The extraction procedures were applied to 10 different soil samples and 2 sludges. The DIN-S4-procedure shows the lowest amounts of mobilized copper. The highest parts are extracted by sodium acetate solution. For the procedures investigated, the mobilized amounts extracted by ammonium acetate solution are most comparable with those of the pHstat-procedure. The simple sodium acetate procedure is useful for screening to select significant samples from a large number of soils. Thus, the high effort caused by the pHstat-procedure can be reduced to a minimum through the application of this screening procedure. The application of this screening, however, is difficult for samples containing large parts of organic compounds, e.g. sewage sludge. These samples show a different extracting behaviour.     

Photometrische Bestimmung der Zytotoxizität von Chemikalien

Two test methods are presented to assess the cytotoxicity of chemicals. On one hand the protozoonTetrahymena pyriformis and on the other the bacteriumPseudomonas aeruginosa is cultured in defined media containing the test chemicals. Test parameter is the quantity of living cells surviving exposure. The conversion of the tetrazolium salt MTT into formazan is equivalent to cell viability. A mixture of detergent and isopropanol is used to dissolve the blue formazan which is not soluble in water, prior to photometric measurement. For cetyltrimethylammonium bromide, benzyldimethyldodecylammonium chloride, dodecylbenzenesulfonic acid sodium salt, potassium dichromate, 4-toluenesulfonic acid sodium salt, and 4-nitrophenol the EC50-values (mol/l) in theTetrahymena pyriformis- (Pseudomonas aeruginosa) test system were: 10^?5,81 (10^?5,08); 10^?4,68) (10^?3,80; 10^?4,50 (10^?4,14); 10^?3,97 (10^?5,11); 10^?2,71 (10^?2,50) and 10^?4,00 (10^?3,64). — With the assays described here it is possible to measure a large number of samples by quick and simple means.     

Berücksichtigung von Variabilität und Unsicherheit in quantitativen Risikoabschätzungen (QRA)

Using the example of residential living on a contaminated site, a probabilistic exposure assessment is performed with variability and uncertainty being modelled separately. Probability distributions are used in the exposure model in order to characterize person-related variables (e.g. body weight) only; chemical-specific parameters are being held constant. In addition, uncertainty concerning one selected variable (soil ingestion rate) was modelled. Comparing these results to conventional “worst case” estimates, we find those estimates located in the uppermost range of the probabilistic estimates. The worst case estimates tend to be highly conservative and possibly unrealistic.     

Entwicklung und Einsatz eines standardisierten Labortests für Dungkäfer (Aphodius constans) zur Prüfung der Ökotoxizität von Veterinärpharmaka

Goal, Scope, and Background

According to European legislation, an environmental risk assessment of veterinary pharmaceuticals for dung beetles is required if the substance is a parasiticide for the treatment of pasture animals. However, the demonstration of the environmental safety of those substances for dung beetles is strongly hampered by the fact that no standardized test system is available so far. Therefore, starting with recommendations from the SETAC advisory group DOTTS (Dung Organism Toxicity Test Standardization), a test system using the widespread temperate dung beetle species Aphodius constans was developed.

Methods

Using experiences gained at the University of Montpellier (e.g. in field studies) and in other terrestrial tests, details of culturing and testing of A. constans were investigated in a project sponsored by the German Environment Protection Agency (UBA). Basically, the survival of young larvae of A. constans exposed to the test substance spiked into formulated (i.e. dried, grounded and re-wetted) or fresh dung is measured over a period of three weeks. Using the model substance dimethoate (an insecticide required as positive control in tests with the predatory mite Hypoaspis aculeifer) the suitability of the new test was checked. Afterwards, the new test method was used to determine the toxicity of four common veterinary pharmaceuticals (ivermectin, dicyclanil, moxidectin, praziquantel) in formulated and fresh dung. Finally, the effects of ivermectin on the dung beetles were determined in a test using fresh dung from treated cattle.

Results and Conclusions

For Dmethoate LC₅₀-values were found in a range of 1–3 mg a.s./kg dung (dew.), depending on the dung type. Praziquantel did not show eirecis in the tested concentration range (i.e. up to 1000 mg a.s./kg Dung (DW). The LC₅₀-values of the other three pharmaceuticals were very low (<6 mg a.s./kg dung (DW)). With the exception of Dicyclanil no differences between formulated and fresh dung were identified. In the case of ivermectin the LC₅₀-value were even lower: about 0.5 mg a.s./kg dung (DW). No differences between the results of test in which the substance was spiked into dung and those tests which were performed with dung from treated cattle were found. These results prove the sensitivity as well as the robustness of this test method.

Recommendations and Perspectives

Based on the experiences reported here, it is recommended to standardize this test in an international ring test so that it can be incorporated in the risk assessment process for veterinary pharmaceuticals as described in the currently developed international registration guidelines.     

Bestimmung von leicht freisetzbarem Cyanid in Böden und Wässern mit der Mikrodestillation

Easy-liberatable cyanide has to be analysed if soils and waters are contaminated by cyanide. The aim of this study was to determine easy-liberatable cyanide in these environmental samples using a micro-distillation apparatus by means of a modified digestion vessel. Pure aqueous solutions of different cyanide species, five contaminated and uncontaminated soil samples and two water samples were analysed by micro-distillation according to the German standards. Recovery was determined by analysis of spiked samples. When using the modified digestion vessels, the pH of 4 which is demanded by the standard method can be adjusted. The recovery of potassium cyanide and weakly-complexed zinc-cyanide ranged from 93–101% for standards, and from 87–98% for spiked samples. In contrast, the recovery of strong iron-cyanide complexes was below 4% both for pure solutions and spiked samples. The precision of the method expressed as a relative standard deviation was 25% in cases of very low easy-liberatable cyanide contents (< 1 mg CN kg^?1) and below 12% in case of high easy-liberatable cyanide contents (> 1 mg CN kg^?1) for contaminated soils. The determination of easily-liberatable cyanide in soils and waters using micro-distillation combined with the modified digestion sample is an alternative to other distillation methods.     

Verteilung und Abnahme von bodennahem Ozon in städtischen und ländlichen Regionen

The compilation of ozone data for the federal states of Hesse and Northrhine-Westphalia (NRW) in Germany indicated that the concentration of ozone level slightly decreased during the years 1990–1998. The average concentration of ozone over forest areas is significantly higher than over cities. Only the maximum figures in the years approached one another. However, values passing the legal thresholds (180 μg ozone/m³) were two to three-fold higher over forests than over cities. The ozone concentration in air is inversely proportional to the traffic density. It is suggested that the lower NOx concentration over the forest than over cities is involved in the maintenance of the higher ozone-concentration over forest areas. In the cities, the ozone is reduced by NO to almost zero at night, whereas it is reduced by only about 50% over forests with lower NO concentrations. This reduction is only partially compensated in connection with the photolysis of NO₂ and the subsequent oxidation of O₂ to O₃ during the day. The ozone situation is principally the same in the federal states of Hesse and Northrhine-Westphalia.     

Identifizierung toxischer Verbindungen in Sedimenten: Ansätze zur Integration von Wirkung und Bioverfügbarkeit

Sediments play an important role for aquatic ecosystem functions. However, they also act as sink, storage and source of lipophilic toxicants and metals. Effect-directed analysis (EDA) is a powerful tool to identify compounds causing adverse effects. In order to avoid misinterpretations and biased prioritization bioavailability needs to be considered together with effects. Bioavailability is a complex process finally resulting in the transfer of a so far particle bound molecule to the target location within the organism where it causes an effect. In order to operationalize this concept for EDA it can be divided into several partial processes. These include desorption from sediments and thus bioaccessibility, equilibrium partitioning of desorbable compounds between sediment, water and organisms driven by activity, and toxicological bioavailability as a result of toxicokinetics including resorption, transport, metabolization and excretion. Bioaccessibility is based on desorption kinetics and can be simulated with mild extraction methods e.?g. using TENAX. Equilibrium partitioning can be simulated with partition-based dosing techniques. First results with these approaches indicate that consideration of bioavailability increases the significance of polar sediment-associated toxicants relative to classical non-polar contaminants such as polycyclic aromatic hydrocarbons.     

Ansätze zur regionalen Quantifizierung von Methan aus Pflanzen

Background

Methane as a gas influencing our climate is of high importance according to the Kyoto-Protocol. The more precisely we can determine anthropogenic and biogenic sources, the more effective measures we can take to reduce the gas. Direct emission of methane from plants apparently contributes significantly to the total emission but has not been accounted for in emission balances until now.

Methods

A model was created to calculate methane emission for a short time period in regional resolution for Lower Austria and to relate the results to known literature.

Results and Discussion

First model results suggest that the amount of methane released from plants in relation to emissions from other known sources are of importance.

Perspectives

The uncertainties associated with both measurements and quantification require reduction by further research.     

Untersuchung von Abfällen mit biologischen Testverfahren zur Bewertung der Wassergefährdung

Wastes from three different types of waste treatment facilities were examined with bioassays to determine their hazard potential to waters (→Part I). All examined wastes showed toxic effects and have to be classified as hazardous to waters according to section 19g of the German Federal Water Management Act. The toxicity is probably caused by heavy metals in the leachates, especially copper. An evaluation pattern to classify wastes in the German system of Water Hazard Classes is presented. According to this proposal, a classification of the examined wastes into Water Hazard Class 1 seems to be appropriate. This classification does not describe the hazards resulting from the regular disposal on a landfill or from the utilisation of the treated wastes.     

Bestimmung von Nitrodiphenylaminen und verwandten Verbindungen im Sickerwasser einer Rüstungsaltlast

Goal, Scope and Background

Nitrodiphenylamines can be found at abandoned military sites where the explosive 2,2′,4,4′,6,6′-hexanitrodiphenylamine (hexyl) or diphenylamine as a stabilizer of smokeless powder was handled. Aim of the present study is the development of HPLC methods for the analysis of contaminants which can be expected in particular under aerobic conditions at premises which are contaminated by nitrodiphenyl-amines and related compounds.

Methods

A list of 13 compounds which can be expected at these explosives factories was deduced from literature studies. HPLC methods were developed for the analytes. Water samples from the unsaturated zone of a former hexyl-producing factory were collected by suction cups and the material was analysed according to these procedures to demonstrate the practicability of the new methods and to verify the existence of the postulated compounds in the environment of the former nitration plant.

Results and Conclusion

The new HPLC methods are suitable for the exploration of sites contaminated by nitrodiphenylamines. Beside some hexyl its intermediates 1-chloro-2,4-dinitrobenzene, 2,4-dinitrodiphenylamine und 2,2′,4,4′-tetranitrodiphenylamine were identified in the leachate water.

Recommendation and Perspective

It is advisable to include at least these 4 compounds in the examination of former hexyl-producing plants. Several unknown peaks were observed in the HPLC-chromatogram. It is recommendable to perform further investigations of the unidentified compounds to compile a final list of analytes for military sites polluted by nitrodiphenylamines.     

Das Biologische System der Elemente (BSE): Eine modelltheoretische Betrachtung zur Essentialität von chemischen Elementen

The correlation of data on the elements obtained by instrumental chemical analysis of highly representative plant samples led to the establishment of a first Biological System of Elements (BSE) in 1994. Including other physiological parameters like availability of chemical elements, transport, storage, etc. it is clear, that aside from the bioinorganic aspects of coordination chemistries of given elements, features of essentiality or toxicity depend on their interference with enzymatic processes. Notably, the latter are not to be considered as purely catalytic transformations, but are instead related to the reproduction of organisms in a direct or indirect manner — as autocatalytic: a process whereby a protein containing some metal or burdened in function by its presence is indirectly involved in its own reproduction. Stoichiometric Network Analysis (SNA) explicitly deals with the general dynamics of such autocatalytic systems. Given there is a relationship between the kinds of metal or metalloid species and the key biological/biochemical transformations to be promoted using them — a relationship which is the topic of bioinorgnaic chemistry-, and that biochemistry is in effect about systems which can reproduce and thus behave autocatalytically, one can expect SNA to yield statements on the basic features of biology and biochemistry as well.