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On-line estimation of biodegradation in an unsaturated soil.

The objective of this study was to develop a model-based estimator of biodegradation in unsaturated soil. This would allow real-time assessment of the efficiency of treatment bioprocesses, such as bioventilation and biopile, and eventually permit optimization through the implementation of control strategies. Based on a reduced-order model, an asymptotic observer was designed to estimate on-line the contaminant concentration, using carbon dioxide measurement. Two observer-based estimators were built to approximate: (1) the specific microbial growth rate; and (2) the biocontact kinetics representing the soil resistance to contaminant biodegradation. State observers and parameter estimators were confronted with the experimental results of biodegradation in microcosms. Hexadecane was used as the model compound, representing petroleum hydrocarbons. Three water contents, corresponding to 20%, 50% and 80% of the water-holding capacity, were tested. The asymptotic observer is able to predict hexadecane depletion with an error on the overall time trajectories of 13%, 8% and 4% for the dry, intermediate and wet soils, respectively, which is acceptable given that all the biokinetic parameters were identified from a biodegradation experiment in liquid phase. The observer-based estimator of the specific microbial growth rate, based on the CO(2) measurement, was successfully calibrated using the off-line measurements of hexadecane as validation data, and allowed estimation of the time when biodegradation switched from a microbial to a biocontact limitation. The biocontact kinetics was also identified on-line, using an estimator based on the hexadecane not in biocontact. These results are very encouraging with respect to the potential for on-line assessment of the performance of treatment bioprocesses in unsaturated soils.

Alkanes↗

Contribution of the Earthworm Lumbricus rubellus (Annelida, Oligochaeta) to the Establishment of Plasmids in Soil Bacterial Communities.

The contribution of the earthworm Lumbricus rubellus in spreading plasmids from a nonindigenous bacterial species to the soil microbial community was studied with Escherichia coli strains as donor organisms. The selected donor strains harbored marker-gene tagged plasmids with different transfer properties and host ranges. Prototrophic benzoate degrading indigenous bacteria were analyzed as potential recipients. In filter-mating experiments, donor strains were mixed with bacterial cell consortia extracted from earthworm casts (feces) and incubated on nutrient agar at 28 degrees C. Transfer was detected with the broad host range IncP plasmid pRP4luc; with the IncQ plasmid, pSUP104luc, but only when it was present in a mobilizing donor strain; and with the transposon delivery vector pUTlux. No transfer was detected with the nonmobilizable pUCluc and the mobilizable pSUP202luc, both of narrow host range. In microcosm studies with E. coli inoculated soil incubated at 12 degrees C, transconjugants were only detected in casts of L. rubellus but not in bulk soil, indicating that the gut passage was a precondition for plasmid transfer. Plasmid pRP4luc was transferred at higher frequencies than detected in filter mating. Results of the filter matings were confirmed except that transfer of pUTlux could not be detected. The majority of transconjugants isolated in this study lost their acquired plasmid upon further cultivation. Stable transconjugants, however, were obtained and identified at the 16S rRNA gene level as members of the b- and g-subgroups of Proteobacteria. Incubation of E. coli and selected transconjugants in soil microcosms with L. rubellus demonstrated that the gut passage resulted in a slight but significant reduction of ingested cells. In contrast to the donor strains, however, the population sizes of transconjugants in bulk soil and in casts did not decrease over time. This demonstrated that the transferred plasmids had established themselves in the soil microbial community.

Journal Article↗

Implementing community-based education: essential elements and recommendations.

Two elements are essential for implementing community-based educational programs: a vision of how community experiences fit into the training of the health professionals of the future and a local environment that will support innovation, change, and growth. Change cannot occur unless very basic assumptions regarding medical education are challenged. What is "quality education"? Can programs oriented toward tertiary and specialty care adequately provide the training that should be the core of a 3-year general training program? Do schools and programs select and train physicians to function within the microcosm of the academic center, or do they prepare physicians to manage the country's health care needs? National consensus has had little influence over local environments. Each medical school, training program, and teaching facility must reexamine its values and its culture. Each must have a vision of the physicians of the future and a commitment to train them appropriately. The role of vision and culture in creating successful programs has been clearly described. Several key elements have been found to be consistent with success, the first being vision. Four basic principles will bring the vision to fruition: (1) preserving core values while still stimulating progress; (2) emphasizing the process by which programs are created, implemented, and changed rather than the product; (3) avoiding the "tyranny of the or," learning to be inclusive with a broad vision rather than limited to an "either-or" approach; and (4) aligning the process, management, and values in working toward envisioned progress for the future. Effective leadership is essential for a group or organization to accomplish its mission, as is an organizational structure that aligns responsibility, authority, resources, and accountability.

Community Medicine↗

The ecology of energy and nutrient fluxes in hemlock forests invaded by hemlock woolly adelgid.

The hemlock woolly adelgid (HWA, Adelges tsugae Annand) is currently causing a severe decline in vitality and survival of eastern hemlock in North American forests. We analyzed the effects of light HWA infestation on vertical energy and nutrient fluxes from the canopy to the forest floor. Canopy throughfall, litter lysimeters, and laboratory litter microcosms were used to examine the effects of HWA-affected and unaffected throughfall on litter type, leachate, and litter chemistry. Early in the season adelgid infestation caused higher dissolved organic carbon (DOC; +24.6%), dissolved organic nitrogen (DON; +28.5%), and K (+39.3%) fluxes and lower inorganic nitrogen fluxes (-39.8%) in throughfall and in adjacent litter solutions collected beneath infested compared to uninfested trees. Needle litter collected beneath uninfested hemlock had significantly lower N concentrations compared to needles collected beneath infested trees, while no difference in N concentrations was found in birch litter. Bacteria were significantly more abundant on hemlock and birch litter beneath infested trees, while yeasts and filamentous fungi showed no consistent response to HWA throughfall. Litter microcosms showed that less DOC was leaching from birch than from hemlock needles when exposed to HWA throughfall. Overall, NH4-N and DON leachate concentrations were higher from birch than from hemlock litter. Thus, HWA-affected throughfall leads to qualitative and quantitative differences in nitrogen export from the litter layer. The N concentration of hemlock litter did not change with time, but the N concentration in birch litter increased significantly during the course of the experiment, especially when HWA-affected throughfall was applied. We suggest a nonlinear conceptual model for the temporal and vertical transition of energy and nutrient fluxes relative to progressing HWA infestation from a pure hemlock to a birch/maple-dominated forest. Progressive needle loss and changes in needle chemistry are likely to produce a humped-shaped DOC curve, while N fluxes initially decrease as infestation continues but rise eventually with hemlock decline and immigration of hardwood species. These findings suggest that it is necessary to understand the biology and specific physiological/trophic effects of exotic pests on their hosts and associated ecosystem processes in order to decipher the temporal dynamics, direction of change, and functional consequences.

Animals↗

The shape of public health policy: the Australian experience.

Public health policy is shaped by many factors. A brief historical reflection is given on policy development in Australia to illustrate the various influences on health policy. Medical technology; ethical trade-offs; environmental, social, and political imperatives; popular movements; and changing patterns of disease; as well as market forces have helped to shape Australian contemporary public health policy. These multiple and often competing forces, however, can work against individual consumer choice in health care decisions. This article demonstrates through the eyes of history the factors that shape public health policy. As Australia has a short history compared to most industrialized democratic societies and can be viewed as a microcosm, it is used as the exemplar.

Australia↗

Comparative oxidation and net emissions of methane and selected non-methane organic compounds in landfill cover soils.

The surface emissions of methane (CH4) and non-methane organic compounds (NMOCs) were determined at two different areas at a French landfill: a permanently covered and fully vegetated area (40 cm coarse sand + 80 cm of loam) and a temporarily covered area (40 cm of coarse sand). The 37 NMOCs quantified in the landfill gas samples included alkanes (C1-C10), alkenes (C1-C4), halogenated hydrocarbons (including (H)CFCs), and aromatic hydrocarbons. Both positive and negative CH4 fluxes ranging from -0.01 to 0.008 g m(-2) d(-1) were measured from the permanently covered cell. However, high spatial variation was observed, and a hot spot with a high flux (10 g m(-2) d(-1)) was identified. A higher CH4 emission occurred from the temporarily covered cell (CH4 flux of 49.9 g m(-2) d(-1)) as compared to the permanently covered cell. The NMOC fluxes from the permanently covered zone were all very small with both positive and negative fluxes in the order of 10(-7) to 10(-5) g m(-2) d(-1). Higher and mainly positive NMOC fluxes in the order of 10(-5) to 10(-4) g m(-2) d(-1) were obtained from the temporarily covered zone. The lower emission from the permanently covered and fully vegetated cell was attributable to the thicker soil layer, which functions as microbial habitat for methanotrophic bacteria. The NMOC oxidation capacity was investigated in soil microcosms incubated with CH4. Maximal oxidation rates for the halogenated aliphatic compounds varied between 0.06 and 8.56 microg (g of soil)(-1) d(-1). Fully substituted hydrocarbons (tetrachloromethane, perchloroethylene, CFC-11, CFC-12, and CFC-113) were not degraded in the presence of CH4 and O2. Benzene and toluene were rapidly degraded, giving very high maximal oxidation rates (28 and 39 microg (g of soil)(-1) d(-1)). On the basis of the emission measurements and the batch experiments conducted, a general pattern was observed between emissions and biodegradability of various NMOCs. The emissions mainly consisted of compounds that were not degradable or slowly degradable, while an uptake of easily degradable compounds was registered. As an example, perchloroethylene, trichloromethane, CFC-11, and CFC-12 were emitted, while atmospheric consumption of aromatic hydrocarbons and lower chlorinated hydrocarbons such as vinyl chloride, dichloromethane, and chloromethane was observed. This study demonstrates that landfill soil covers show a significant potential for CH4 oxidation and co-oxidation of NMOCs. Under certain conditions, landfills may even function as sinks for CH4 and selected NMOCs, like aromatic hydrocarbons and lower chlorinated compounds.

Air Pollution↗

Gene transfer in marine water column and sediment microcosms by natural plasmid transformation.

We investigated the possibility for natural transformation in the marine environment by using broad-host-range plasmid multimers and a high-frequency-of-transformation (HFT) Vibrio strain as the recipient. Water and sediment samples were taken from Tampa Bay, the eastern Gulf of Mexico, the Florida Shelf near Miami, and the Bahamas Bank. In water column microcosms, transformation frequencies ranged from 1.7 x 10 to 2.7 x 10 transformants per recipient, with highest frequencies occurring when low levels of nutrients (peptone and yeast extract) were added. The presence of the ambient community either reduced transformation frequency by an order of magnitude or had no effect. In sterile sediments, nutrient additions had no consistent effect on transformation, with transfer frequencies similar to those observed in the water column. Transformation was not observed in any sediment experiment when the ambient microbial community was present. These findings are the first report of natural plasmid transformation in seawater and in the presence of the ambient microbial community. This process may be a mechanism for the acquisition of small, nonconjugative plasmids, which are commonly found in aquatic bacteria. Our data also suggest that natural transformation may be more likely to occur in the water column than in native marine sediments, contradicting prior conclusions based on studies with sterile sediments.

Journal Article↗

Characterization and identification of numerically abundant culturable bacteria from the anoxic bulk soil of rice paddy microcosms.

Most-probable-number (liquid serial dilution culture) counts were obtained for polysaccharolytic and saccharolytic fermenting bacteria in the anoxic bulk soil of flooded microcosms containing rice plants. The highest viable counts (up to 2.5 x 10(8) cells per g [dry weight] of soil) were obtained by using xylan, pectin, or a mixture of seven mono- and disaccharides as the growth substrate. The total cell count for the soil, as determined by using 4', 6-diamidino-2-phenylindole staining, was 4.8 x 10(8) cells per g (dry weight) of soil. The nine strains isolated from the terminal positive tubes in counting experiments which yielded culturable populations that were equivalent to about 5% or more of the total microscopic count population belonged to the division Verrucomicrobia, the Cytophaga-Flavobacterium-Bacteroides division, clostridial cluster XIVa, clostridial cluster IX, Bacillus spp., and the class Actinobacteria. Isolates originating from the terminal positive tubes of liquid dilution series can be expected to be representatives of species whose populations in the soil are large. None of the isolates had 16S rRNA gene sequences identical to 16S rRNA gene sequences of previously described species for which data are available. Eight of the nine strains isolated fermented sugars to acetate and propionate (and some also fermented sugars to succinate). The closest relatives of these strains (except for the two strains of actinobacteria) were as-yet-uncultivated bacteria detected in the same soil sample by cloning PCR-amplified 16S rRNA genes (U. Hengstmann, K.-J. Chin, P. H. Janssen, and W. Liesack, Appl. Environ. Microbiol. 65:5050-5058, 1999). Twelve other isolates, which originated from most-probable-number counting series indicating that the culturable populations were smaller, were less closely related to cloned 16S rRNA genes.

Alphaproteobacteria↗

Unparallel diversification in bacterial microcosms.

Adaptive speciation has gained popularity as a fundamental process underlying the generation of diversity. We tested whether populations respond to similar forms of disruptive selection by diversifying in similar or parallel ways by investigating diversified populations of Escherichia coli B evolved in glucose and glucose-acetate environments. In both environments, the populations have differentiated into two phenotypes, named for their characteristic colony morphologies: large (L) and small (S). Each type is heritable and this polymorphism (or 'diversified pair') appears to be maintained by negative frequency dependence. The L and S phenotypes from different environments are convergent in their colony morphology and growth characteristics. We tested whether diversification was parallel by conducting competition experiments between L and S types from different environments. Our results indicate that replicate diversified pairs from different environments have not diversified in parallel ways and suggest that subtle differences in evolutionary environment can crucially affect the outcome of adaptive diversification.

Acetates↗

A study of the degradation of phenoxyacid herbicides at different sites in a limestone aquifer.

The biodegradation of three phenoxyalkanoic acid herbicides, viz. MCPA, dichlorprop and mecoprop, under aerobic and anaerobic conditions was investigated using microcosm techniques. The field studies were conducted in a limestone aquifer that had suffered contamination from leaking landfill sites in which phenoxyalkanoic acid herbicides (mostly mecoprop) had undergone disposal. The results from in situ field and laboratory microcosms indicated that under microbially active aerobic conditions the biotransformation of all three herbicides was rapid and that lag phases were short. Under fully aerobic conditions the concentration of each of the three herbicides was reduced from 2,000 micrograms/l to below the detection limit (approx. 10 micrograms/l) of the HPLC system, used for their analysis, within 14 days. However, under microbially active anaerobic conditions no degradation of the herbicides could be discerned over the 100-200 day duration of the experiments. This finding has significant implications for the disposal of phenoxyalkanoic acid herbicides particularly in situations where any resulting leachate may find its way into underlying water resources.

2,4-Dichlorophenoxyacetic Acid↗

Persistence of testosterone and 17beta-estradiol in soils receiving swine manure or municipal biosolids.

Natural and synthetic steroidal hormones can be carried to agricultural soil through fertilization with municipal biosolids, livestock manure, or poultry manure. The persistence and pathways of dissipation of [4-(14)C]-testosterone and of [4-(14)C]-17beta-estradiol in organic-amended soils were investigated using laboratory microcosms. Testosterone dissipation was investigated over a range of amendment concentrations, temperatures, and soil types. Under all conditions the parent compound and transformation products were dissipated within a few days. Addition of swine manure slurry to soil hastened the transformation of testosterone and 17beta-estradiol to the corresponding less hormonally active ketones, 4-androstene-3,17-dione and estrone. Two other testosterone transformation products, 5alpha-androstan-3,17-dione and 1,4-androstadiene-3,17-dione, were also detected. Experiments with sterilized soil and sterilized swine manure slurry suggested that the transformation of (14)C-labeled hormonal parent compounds was mainly caused by microorganisms in manure slurry, while mineralization of the hormones to (14)CO(2) required viable soil microorganisms. Organic amendments transiently inhibited the mineralization of [4-(14)C]-testosterone, perhaps by inhibiting soil microorganisms, or by enhancing sorption and reducing the bioavailability of testosterone or transformation products. Overall, organic amendments influenced the pathways and kinetics of testosterone and estradiol dissipation, but did not increase their persistence.

Animals↗

Effects of atrazine and nicosulfuron on phytoplankton in systems of increasing complexity.

We have tested the sensitivity of phytoplankton to the herbicides atrazine and nicosulfuron in experiments conduced in increasingly complex systems, from single strain phytoplankton cultures (microplates) to mesocosms mimicking whole ecosystems. The endpoints used to assess sensitivity to atrazine and nicosulfuron were total biomass increase, photosynthetic efficiency, and community diversity, depending on the system considered. Nicosulfuron appeared to be very much less toxic to phytoplankton than atrazine, in accord with the planned changes in agricultural practices to reduce the effects of surface water contamination on aquatic biota. Nevertheless, nicosulfuron had significant effects in some systems (principally microcosms), whereas the single monocultures were almost insensitive to it. This points out the inaccuracy of standardized toxicity test on phytoplanktonic algae alone for predicting the effects of xenobiotics on natural communities and the need for tests in microcosms and mesocosms to obtain reliable evidence about the toxicity of a given chemical on freshwater aquatic ecosystems.

Atrazine↗

Ecological effects of spring and late summer applications of lambda-cyhalothrin on freshwater microcosms.

The aim of the study was to compare the effects of the pyrethroid insecticide lambda-cyhalothrin (treated at 10, 25, 50, 100, 250 ng active ingredient a.i./L) on a drainage ditch ecosystem in spring and late summer. Microcosms (water volume approximately 430 L) were established using enclosures in a 50-cm-deep experimental ditch system containing communities typical of macrophyte-dominated freshwater ecosystems. Effects on macroinvertebrates, zooplankton, phytoplankton, macrophytes, and community metabolism were assessed and evaluated using univariate and multivariate statistical techniques. The macroinvertebrate community responded most clearly to treatment and, as anticipated, insects and crustaceans were among the most sensitive organisms. Statistical analysis showed that the underlying community structure was significantly different between the spring and summer experiments. However, the most sensitive species (Chaoborus obscuripes and Gammarus pulex) were abundant in spring as well as in late summer. In spring and late summer, only slight and transient effects were observed at the community level in the 10-ng/L treatment. Overall, the study did not show substantial differences in the responses of sensitive taxa between spring and late summer treatments, and effects thresholds were similar irrespective of season of treatment.

Animals↗

Biodegradation of 1,4-dioxane in planted and unplanted soil: effect of bioaugmentation with Amycolata sp. CB1190.

1,4-Dioxane is one of the most recalcitrant and toxic contaminants in the subsurface. This study investigated the potential to enhance dioxane biodegradation in both planted and unplanted soil, by adding the dioxane-degrading actinomycete, Amycolata sp. CB1190. Dioxane was not removed within 120 days in sterile controls or in viable microcosms not amended with CB 1190. Poplar root extract (40 mg/L as COD) stimulated dioxane degradation in bioaugmented soil, and 100 mg/L dioxane were removed within 45 days. Other co-substrates that enhanced dioxane degradation by CB1190 include tetrahydrofuran (THF) and 1-butanol, while glucose and soil extract did not affect dioxane degradation. The stimulatory effect of THF was partly due to enhanced enzyme induction, while that of root extract and 1-butanol was attributed to additional growth of CB1190. In another experiment with dioxane added at 10 mg/kg-soil. reactors planted with hybrid poplar trees removed (by evapotranspiration and biodegradation in the root zone) more dioxane within 26 days than unplanted reactors, regardless of whether CB1190 was added. Nevertheless, CB1190 enhanced mineralization of [14C]-dioxane in all experiments. This enhancement was more pronounced in unplanted soil because plant uptake reduced the availability of dioxane for microbial degradation. These results suggest that bioaugmented phytoremediation is an attractive alternative to remove dioxane from shallow contaminated sites.

Actinomycetales↗

Effects of mercury contamination on the culturable heterotrophic, functional and genetic diversity of the bacterial community in soil.

This study investigates the effect of mercury contamination on the culturable heterotrophic, functional and genetic diversity of the bacterial community in soil. The changes in diversity were monitored in soil microcosms, enriched with 25 &mgr;g Hg(II) g(-1) soil, over a period of 3 months. The culturable heterotrophic diversity was investigated by colony morphology and colony appearance on solid LB medium. Functional diversity was analysed as sole carbon utilisation patterns in ECOplates. Genetic diversity was measured as bands on denaturing gradient gel electrophoresis (DGGE) gels obtained by purification of total soil DNA and amplification of bacterial 16S rDNA fragments by polymerase chain reaction. Concentrations of bioavailable and total mercury were measured throughout the experiment. The effect on the culturable heterotrophic and genetic diversity was very similar, showing an immediate decrease after mercury addition but then slowly increasing throughout the entire experimental period. Pre-exposure levels were not reached within the time span of this investigation. The DGGE band pattern indicated that a shift in the community structure was responsible for recovered diversity. When analysed by Shannon-Weaver indices, functional diversity was found to increase almost immediately after mercury addition and to remain at a level higher than the control soil for the rest of the experiment. The fraction of culturable heterotrophic bacteria increased from 1% to 10% of the total bacterial number as a result of mercury addition, and the mercury-resistant population increased to represent the entire heterotrophic population.

Journal Article↗

Oxygen-sensing reporter strain of Pseudomonas fluorescens for monitoring the distribution of low-oxygen habitats in soil.

The root-colonizing bacterium Pseudomonas fluorescens CHA0 was used to construct an oxygen-responsive biosensor. An anaerobically inducible promoter of Pseudomonas aeruginosa, which depends on the FNR (fumarate and nitrate reductase regulation)-like transcriptional regulator ANR (anaerobic regulation of arginine deiminase and nitrate reductase pathways), was fused to the structural lacZ gene of Escherichia coli. By inserting the reporter fusion into the chromosomal attTn7 site of P. fluorescens CHA0 by using a mini-Tn7 transposon, the reporter strain, CHA900, was obtained. Grown in glutamate-yeast extract medium in an oxystat at defined oxygen levels, the biosensor CHA900 responded to a decrease in oxygen concentration from 210 x 10(2) Pa to 2 x 10(2) Pa of O(2) by a nearly 100-fold increase in beta-galactosidase activity. Half-maximal induction of the reporter occurred at about 5 x 10(2) Pa. This dose response closely resembles that found for E. coli promoters which are activated by the FNR protein. In a carbon-free buffer or in bulk soil, the biosensor CHA900 still responded to a decrease in oxygen concentration, although here induction was about 10 times lower and the low oxygen response was gradually lost within 3 days. Introduced into a barley-soil microcosm, the biosensor could report decreasing oxygen concentrations in the rhizosphere for a 6-day period. When the water content in the microcosm was raised from 60% to 85% of field capacity, expression of the reporter gene was elevated about twofold above a basal level after 2 days of incubation, suggesting that a water content of 85% caused mild anoxia. Increased compaction of the soil was shown to have a faster and more dramatic effect on the expression of the oxygen reporter than soil water content alone, indicating that factors other than the water-filled pore space influenced the oxygen status of the soil. These experiments illustrate the utility of the biosensor for detecting low oxygen concentrations in the rhizosphere and other soil habitats.

Bacterial Proteins↗

Polyhydroxyalkanoate degradation is associated with nucleotide accumulation and enhances stress resistance and survival of Pseudomonas oleovorans in natural water microcosms.

Pseudomonas oleovorans GPo1 and its polyhydroxyalkanoic acid (PHA) depolymerization-minus mutant, GPo500 phaZ, residing in natural water microcosms, were utilized to asses the effect of PHA availability on survival and resistance to stress agents. The wild-type strain showed increased survival compared to the PHA depolymerase-minus strain. The appearance of a round cellular shape, characteristic of bacteria growing under starvation conditions, was delayed in the wild type in comparison to the mutant strain. Percent survival at the end of ethanol and heat challenges was always higher in GPo1 than in GPo500. Based on these results and on early experiments (H. Hippe, Arch. Mikrobiol. 56:248-277, 1967) that suggested an association of PHA utilization with respiration and oxidative phosphorylation, we investigated the association between PHA degradation and nucleotide accumulation. ATP and guanosine tetraphosphate (ppGpp) production was analyzed under culture conditions leading to PHA depolymerization. A rise in the ATP and ppGpp levels appeared concomitant with PHA degradation, while this phenomenon was not observed in the mutant strain unable to degrade the polymer. Complementation of the phaZ mutation restored the wild-type phenotype.

Biodegradation, Environmental↗

Isotopic fractionation indicates anaerobic monochlorobenzene biodegradation.

The concentration and isotopic composition of monochlorobenzene (MCB) was monitored in the plume of an anaerobic, contaminated aquifer in Bitterfeld, Germany. An enrichment in the carbon isotopic composition of more than 4 delta units was found at the fringes of the plume relative to the center (-26.5 %), suggesting the occurrence of in situ biodegradation of MCB. A similar enrichment was measured in a detailed cross-section of the plume and in depth-specific samples obtained in a multilevel sampling well. The latter samples gave a good correlation of MCB concentrations and respective isotopic composition according to the Rayleigh equation. On the other hand, batch experiments using the aerobic MCB-degrading strains Ralstonia sp. DSM 8910, Acidovorax facilis UFZ B517, Rhodococcus erythropolis UFZ B528, and Pseudomonas veronii UFZ B547 showed that the known aerobic pathway initiated by dioxygenases does not result in a significant isotopic fractionation. Thus, a novel anaerobic pathway resulting in an isotopic fractionation appears to be the predominant process of MCB degradation in this aquifer. The study also clearly demonstrates the usefulness of isotopic fractionation analysis to prove biodegradation directly in the field, even when microcosm studies are not available and a metabolic pathway has not yet been elucidated.

Anaerobiosis↗