Mikrobielle Sanierung von 2,4,6-Trinitrotoluol (TNT) kontaminierten Böden

Within the limits of a feasability study abouton-site bioremediation methods for TNT-contaminated soils, composting was chosen as a very promising and cheap method. This method was critically compared with those described in the literature and was primarily rated under ecotoxicological aspects. The investigated location is the former munition plant «Tanne» in the aerea of Clausthal-Zellerfeld in Lower Saxony, Germany. To estimate the autochtonic microflora, we assessed the number of aerobic heterotrophic bacteria and determined their respiration activity in soils. In addition, we isolated bacteria and examined their capacity to metabolize TNTin vitro. Both the amount of autochtonic microrganisms (4.7×10⁸ to 1.2×10¹⁰ colony forming units (cfu)/kg dryweight) as well as their respiration activity did not correlate with the concentrations of nitrotoluenes in the soils. With high contaminated soil (20 g TNT/kg dry weight) we carried out a small compost in the range of 10 liters. During 28 days of composting TNT-concentration decrease over 90% and only minor amounts of monoaminodinitrotoluenes were generated. However, an acidic pretreatment of the compost material at the end of the reaction showed that TNT could be partially resolved under these extreme conditions and that an ecotoxicological risk may still exist. Possible changes in the realization of the composting process in order to make sure that the contaminants are savely bound to the humin matrix are discussed.     

Anaerobic incorporation of the radiolabeled explosive TNT and metabolites into the organic soil matrix of contaminated soil after different treatment procedures

Four bioreactor designs were performed to evaluate the level of incorporation of 14C-labeled 2,4,6-trinitrotoluene (TNT) and metabolites into the organic soil matrix of different anaerobically treated contaminated soils. The contaminated soils were amended with molasses slivers (80:20% per weight) as auxiliary substrate to enhance microbial activity. After 5 weeks (bioreactors 1 and 2), 8 weeks (bioreactor 3) and 12 weeks (bioreactor 4) of anaerobic incubation, we determined 41%, 58%, 72%, and 54%, respectively, of the initially applied radioactivity immobilized in various soil fractions. After alkaline hydrolyses of the solvent-extracted soils, low quantities of radiolabel were found in the humic and fulvic acid fractions, whereas the bulk of 14C activity was found to be strongly bound to the humin fraction (solid soil residues). The amounts of solvent extractable radioactivity were 53%, 40%, 16%, and 29% for bioreactors 1, 2, 3, and 4, respectively. The level of TNT transformation at the end of the experiments was within 90-94%. Regarding the results presented in this study, we can assume that there is the possibility of high incorporation levels of TNT metabolites into the soil organic matrix mediated by microbial cometabolism under strictly anoxic conditions.     

TNT in Komposten und Flüssigkultur

The formation of aromatic amines was investigated using a summarized test (NEDA-test) during the composting of 2,4,6-trinitrotoluene (TNT) contaminated soil. In this test, the aromatic amines were diazolated and then coupled to N-1-Naphthyl-ethylenediamine-dihydrochloride (NEDA) to yield an azo dye which can be monitored photometrically. The test was calibrated for known TNT-metabolites with an active amine-group. Liquid samples from composting- and liquid-culture-experiments were analyzed by HPLC for these known metabolites. Moreover, the samples were monitored by the NEDA-test and the expected extinction of the TNT-metabolites found with amine function were extrapolated with the help of calibration curves. It was shown that substantial differences are obvious between the monitored and extrapolated values. After separation into polar and non-polar aromatic amines, it became clear that these differences are made by the polar aromatic amines. Polar aromatic amines, which are not detectable by presently available analytical tests, were generated during the composting of TNT-contaminated soils. Contaminated stagnant water, which was generated during anaerobization of a compost prephase, was treated aerobically for 70 days in a biofermenter. During this treatment TNT and its known metabolites were eliminated almost entirely. Simultaneously, the toxicity in the Lumis Tox-test decreased drastically. In striking contrast, the sum of aromatic amines decreased only to a minor extent. Moreover, the percentage of polar compounds from total amount of aromatic amines increased drastically from 48% to more than 95%. At present, the chemical identification of these polar compounds is still missing and is the object for further research.     

Identification of oxidized TNT metabolites in soil samples of a former ammunition plant

Water extracts of soil samples of the former ammunition plant “Tanne” near Clausthal-Zellerfeld, Lower Saxony, Germany, were investigated for highly polar oxidized 2,4,6-trinitrotoluene (TNT) metabolites. 0.4 to 9.0 mg/kg dry soil 2,4,6-trinitrobenzoic acid (TNBA) and 5.8 to 544 mg/kg dry soil 2-amino-4,6-dinitrobenzoic acid (2-ADNBA) were found. In addition to the oxidized metabolites, TNT, 4- and 2-aminodinitrotoluene (4- and 2-ADNT), and 2,4-dinitrotoluene (2,4-DNT) were extractable with water. Most interestingly, in one sample, 2-ADNBA represented the main contaminant. The origin of the oxidized nitroaromatics is unknown at this time. They might be generated chemically or photochemically. Furthermore, a biological synthesis seems possible.