Two Pilot Plant Reactors Designed for the In Situ Bioremediation of Chlorobenzene-contaminated Ground Water: Hydrogeological and Chemical Characteristics and Bacterial Consortia

The SAFIRA in situ pilot plant in Bitterfeld, Saxonia-Anhalt, Germany, currently serves as the test site for eight different in situ approaches to remediate anoxic chlorobenzene (CB)-contaminated ground water. Two reactors, both filled with originallignite-containing aquifer material, are designed for themicrobiological in situ remediation of the ground water bythe indigenous microbial consortia. In this study, thehydrogeological, chemical and microbiological conditions of theinflowing ground water and reactor filling material are presented,in order to establish the scientific basis for the start of thebioremediation process itself. The reactors were put intooperation in June 1999. In the following, inflow CB concentrationsin the ground water varied between 22 and 33 mg L^-1; achemical steady state for CB in both reactors was reachedafter 210 till 260 days operation time. The sediments werecolonized by high numbers of aerobic, iron-reducing anddenitrifying bacteria, as determined after 244 and 285 days ofoperation time. Furthermore, aerobic CB-degrading bacteria weredetected in all reactor zones. Comparative sequence analysis of16S rDNA gene clone libraries suggest the dominance of Proteobacteria (Comamonadaceae, Alcaligenaceae, Gallionella group, Acidithiobacillus) and members of theclass of low G+C gram-positive bacteria in the reactorsediments. In the inflowing ground water, sequences withphylogenetic affiliation to sulfate-reducing bacteria andsequences not affiliated with the known phyla of Bacteria, were found.     

Untersuchungen zum ökotoxikologischen Schädigungspotenzial und Erosionsrisiko von kontaminierten Sedimenten in staugeregelten Flüssen

Particle-bound pollutants accumulate in river-bottom sediments, a process which results in a significant decrease in the ecotoxicological availability of toxicants for the majority of aquatic organisms. Under normal hydrologic conditions, the release of contaminants from bottom sediments is usually of minor importance. In contrast, flood events may remobilize highly contaminated sediments via in-stream erosion. The objective of this study was to develop a combined ecotoxicological and hydraulic approach to elucidate the ecotoxicological implications associated with the risk of erosion of contaminated sediments. This integrated strategy was applied to the lock-regulated Neckar river in Southern Germany. Both the bottom-sediment cores and suspended matter from two intensive flood events were investigated. Sediment samples below an erosional unconformity showed a sharp increase in the ecotoxicological load. Moreover, it was found that major flood events (HQ₅ and higher) could possibly erode even very old, well-consolidated and highlycontaminated sediments. The suspended matter of the high discharge events investigated (return periods of 15 to 20 years) exerted significantly higher cytotoxicity and mutagenicity than a moderate flood with a 1-year return period. These findings support the conclusion that the observed ecotoxicological effects during major floods may at least in part be due to the in-stream erosion of highly contaminated bottom sediments.     

Dioxinbilanz für Hamburg

The sources and reservoirs of polychlorinated dibenzo-p-dioxins and dibenzofurans (PCDD/PCDF) in the city of Hamburg, Germany, in the year 1992 were identified and quantified combining analytical data of PCDD/PCDF with statistical data on annual fluxes of materials and products. Fluxes and amounts of PCDD/PCDF were estimated for the environmental compartments water, air, and soil. Dioxin emissions originating from industrial, commercial or private activities were also calculated. Together with the analysis of the specific regional nutrition data, the human exposurevia the various pathways could be determined. It could be shown that the total PCDD/PCDF intake of individuals in Hamburg was about the same as the national average. High annual fluxes of PCDD/PCDF connected with the trade of industrial products and the waste management and disposal: system in Hamburg obviously had no influence on the human intake of PCDD/PCDF.     

Changes in toxicity and genotoxicity of industrial sewage sludge samples containing nitro- and amino-aromatic compounds following treatment in bioreactors with different oxygen regimes

Goals, Scope and Background

From 2005, deposition of organic waste will be banned in Sweden. Likewise, in Germany and Austria, similar bans are being planned, and further countries will probably follow. Thus, there is a need to develop new methods and to refine established techniques for sludge management in the whole of the European Union. For this end, there is also an urgent need for appropriate ecotoxicological approaches to elucidate and assess the hazard potential of sewage sludge. Therefore, the present study was designed to assess the capacity of various established sludge treatment methods using different oxygen regimes to degrade recalcitrant nitro-substituted organic compounds and reduce their toxicity. Sewage sludge samples from a wastewater treatment plant in Sweden (Cambrex Karlskoga AB, industrial area Björkborn) receiving wastewater from industries manufacturing pharmaceutical substances, chemical intermediates and explosives were processed with different sludge treatment methods. Among other treatment methods, bioreactors (for anaerobic and aerobic sludge treatment) were used. In the present investigation, a battery ofin vitro bioassays was employed to compare the cytotoxic and genotoxic potentials of different fractions of sludge samples in order to elucidate whether the treatments were suitable to reduce the toxicity of the sludge.

Methods

In order to investigate the cytotoxicity of the extracts of treated and untreated sludge samples, the acute cytotoxicity test with the permanent cell line RTL-W1 was used. Genotoxicity was tested by means of the comet assay (single cell gel electro-phoresis) with RTL-W1cells, and mutagenicity was assessed with the Ames test using the Salmonella typhimurium strains TA98, TA98NR and TA100. Sludge toxicity was tested in different fractions of organic extracts produced by acetone and hexane extractions. The subsequent clean-up procedure (silica gel chro-matography and elution with hexane and dichloromethane) resulted in two fractions, a lipophilic hexane-fraction and a semi-lipophilic dichloromethane-fraction. For the genotoxicity and mutagenicity tests, these fractions were reunited at equal ratios.

Results and Discussion

The acute cytotoxicity test with RTL-Wl cells revealed a high cytotoxic potential for the semi-li-pophilic DM-fractions of all sludge samples with NR₅₀ values (= effective concentration for 50% cell death in the neutral red test) from 8.9 up to 20 mg sludge d.w./ml medium. A low cytotoxic potential for the hexane fractions of the untreated sludge samples (NR₅₀ 400 to < 400 mg sludge d.w./ml medium) was observed, whereas the hexane fractions of the treated sludge samples showed elevated cytotoxicity increasing further with treatment in the bioreactors. The comet assay indicated that three out of eight of the reunited fractions had a significant genotoxic potential. Whereas the genotoxic potential of one sample treated anaerobically was very high with an induction factor of 11.6, a similar sample (taken from the same anaerobic reactor four months later) and one untreated sample showed lower potentials. The samples treated in another anaerobic bioreactor as well as the samples treated aerobically showed no genotoxic potential. Results indicate that aerobic treatment was basically adequate for reducing the genotoxicity of the sludge, whereas anaerobic treatment was only partly useful for reduction of genotoxicity. The Ames test revealed a very high mutagenic potential for the reunited fractions of the untreated sludge samples with strain TA98 (maximum induction factors (IF_max up to 45) and a relatively high potential for one of the samples treated aerobically (S2, IF_max = 18 (TA98, S9-)), thus documenting the suitability of both anaerobic and aerobic treatments to reduce the mutagenicity of the samples, however, with the aerobic treatment being less effective. Conclusions. Overall, none of the microbiological treatments for wastewater sludge in bioreactors was found to be ideal for general toxicity reduction of the sludge samples. Whereas cytotoxicity of the sludge increased or levelled off in most cases following either treatment, genotoxicity both increased or decreased after anaerobic treatment, depending on the specific sample. However, mutagenicity could generally be reduced by anaerobic treatment and, to a lesser degree, by aerobic treatment. Recommendationsand Perspectives. The complex modification of the diverse damage potentials of sludge sample extracts by use of anin vitro biotest battery following treatment for toxicity reduction in bioreactors showed that considerations of different toxicological endpoints is essential for an adequate hazard assessment. Whereas in the case of cytotoxicity reduction, the reactors proved ineffective, mutagenicity could be reduced significantly at least in some cases in this case study.