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.     

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.     

Erratum

Murine monoclonal antibodies (mabs) are produced in either mouse ascites or bioreactors (spinner culture, stirred-tank reactor, airlift reactor, hollow-fiber reactor). Human mabs are produced solely in bioreactors. Encapsulation represents a special technology. Hybridoma cells have to be adapted prior to growth in bioreactors. Of crucial importance is the construction of over-producing cell lines by cell- and gene-technological methods. Manipulated cell lines often produce modified mabs.     

The genetic population structure of the gray mouse lemur (<Emphasis Type="Italic">Microcebus murinus), </Emphasis>a basal primate from Madagascar

The genetic structure of a population is closely connected to fundamental evolutionary processes and aspects of social behavior. Information on genetic structure is therefore instrumental for the interpretation of social behavior and evolutionary reconstructions of social systems. Gray mouse lemurs (Microcebus murinus) are basal primates endemic to Madagascar whose social organization is characterized by solitary foraging at night and communal resting during the day. Conflicting reports about population structure based on behavioral observations led us to examine the genetic structure of one population in detail in order to: (1) identify natural genetic units in this solitary primate, and (2) to test the assumption of current models of primate social evolution that solitary primates are organized in matrilines. DNA was extracted from tissue samples of 85 individuals from Kirindy forest to determine their variability at a 530 bp fragment of the mitochondrial D-loop and at six microsatellite loci. We found that this population was characterized by a great general diversity among mtDNA haplotypes, a pronounced sex difference in mtDNA haplotype diversity and spatial clustering of females with a particular haplotype, but low average relatedness among members of haplotype clusters. Specifically, we identified 13 different haplotypes, which were unevenly distributed among individuals. About 80% of all individuals, most of which were females or juvenile males, shared a single haplotype. Rare haplotypes were almost exclusively represented by single adult males, who apparently migrated into this population. One other haplotype was represented by a small group of females living at one edge of the study area. Microsatellite analysis revealed above-average relatedness among females with overlapping home ranges, as well as no signs of inbreeding, implying that male dispersal results in high levels of gene flow among matrilineal groups. We conclude that gray mouse lemur populations are hierarchically organized in small family units of closely related females that form stable sleeping groups, several of which are connected through a common mtDNA haplotype and form spatially distinct clusters. The presence of such matrilines supports a basic assumption of current models of primate social evolution.     

Sperm usage in honey bees

Sperm usage was investigated in a naturally mated honey bee queen. We collected worker progeny arising from eggs that were laid sequentially during three sampling periods. Paternity was determined by analysis of three polymorphic microsatellite loci, leading to the conclusion that the queen had mated with seven males. Direct analysis of the sperm from the spermatheca revealed no evidence that sperm from additional males was present inside the spermatheca. Frequencies of different subfamilies differed significantly and ranged from 3.8% to 27.3%. In the short term, the frequencies of subfamilies among the eggs laid did not change over time. The frequency of eggs of a particular subfamily was statistically independent of the previous egg's subfamily. Thus, there is no evidence for non-random fine-scale sperm usage, and we estimate the effect of sperm clumping to be less than 6%. We conclude that the sperm is mixed completely inside the queen's spermatheca. Our results suggest that taking brood samples from comb cells next to each other is a statistically correct way of independent sampling of subfamilies at a given time in honey bee colonies. Furthermore, any bias in subfamily frequencies in offspring queens due to sperm usage can be excluded. However, the analyses of progeny samples taken 12 months apart do not allow us to exclude moderate fluctuations of subfamily frequencies in the long-term. Received: 11 August 1997 / Accepted after revision: 14 November 1997