Barbara Morasch

Degradation of o-xylene and m-xylene by a novel sulfate-reducer belonging to the genus Desulfotomaculum

A strictly anaerobic bacterium, strain OX39, was isolated with o-xylene as organic substrate and sulfate as electron acceptor from an aquifer at a former gasworks plant contaminated with aromatic hydrocarbons. Apart from o-xylene, strain OX39 grew on m-xylene and toluene and all three substrates were oxidized completely to CO2. Induction experiments indicated that o-xylene, m-xylene, and toluene degradation were initiated by different specific enzymes. Methylbenzylsuccinate was identified in supernatants of cultures grown on o-xylene and m-xylene, and benzylsuccinate was detected in supernatants of toluene-grown cells, thus indicating that degradation was initiated in all three cases by fumarate addition to the methyl group. Strain OX39 was sensitive towards sulfide and depended on Fe(II) in the medium as a scavenger of the produced sulfide. Analysis of the PCR-amplified 16S rRNA gene revealed that strain OX39 affiliates with the gram-positive endospore-forming sulfate reducers of the genus Desulfotomaculum and is the first hydrocarbon-oxidizing bacterium in this genus.

Quantification of biodegradation for o-xylene and naphthalene using first order decay models, Michaelis-Menten kinetics and stable carbon isotopes.

At a former wood preservation plant severely contaminated with coal tar oil, in situ bulk attenuation and biodegradation rate constants for several monoaromatic (BTEX) and polyaromatic hydrocarbons (PAH) were determined using (1) classical first order decay models, (2) Michaelis-Menten degradation kinetics (MM), and (3) stable carbon isotopes, for o-xylene and naphthalene. The first order bulk attenuation rate constant for o-xylene was calculated to be 0.0025 d(-1) and a novel stable isotope-based first order model, which also accounted for the respective redox conditions, resulted in a slightly smaller biodegradation rate constant of 0.0019 d(-1). Based on MM-kinetics, the o-xylene concentration decreased with a maximum rate of k(max)=0.1 microg/L/d. The bulk attenuation rate constant of naphthalene retrieved from the classical first order decay model was 0.0038 d(-1). The stable isotope-based biodegradation rate constant of 0.0027 d(-1) was smaller in the reduced zone, while residual naphthalene in the oxic part of the plume further downgradient was degraded at a higher rate of 0.0038 d(-1). With MM-kinetics a maximum degradation rate of k(max)=12 microg/L/d was determined. Although best fits were obtained by MM-kinetics, we consider the carbon stable isotope-based approach more appropriate as it is specific for biodegradation (not overall attenuation) and at the same time accounts for the dominant electron-accepting process. For o-xylene a field based isotope enrichment factor epsilon(field) of -1.4 could be determined using the Rayleigh model, which closely matched values from laboratory studies of o-xylene degradation under sulfate-reducing conditions.

Intrinsic biodegradation potential of aromatic hydrocarbons in an alluvial aquifer – Potentials and limits of signature metabolite analysis and two stable isotope-based techniques

Three independent techniques were used to assess the biodegradation of monoaromatic hydrocarbons and low-molecular weight polyaromatic hydrocarbons in the alluvial aquifer at the site of a former cokery (Flémalle, Belgium). Firstly, a stable carbon isotope-based field method allowed quantifying biodegradation of monoaromatic compounds in situ and confirmed the degradation of naphthalene. No evidence could be deduced from stable isotope shifts for the intrinsic biodegradation of larger molecules such as methylnaphthalenes or acenaphthene. Secondly, using signature metabolite analysis, various intermediates of the anaerobic degradation of (poly-) aromatic and heterocyclic compounds were identified. The discovery of a novel metabolite of acenaphthene in groundwater samples permitted deeper insights into the anaerobic biodegradation of almost persistent environmental contaminants. A third method, microcosm incubations with 13C-labeled compounds under in situ-like conditions, complemented techniques one and two by providing quantitative information on contaminant biodegradation independent of molecule size and sorption properties. Thanks to stable isotope labels, the sensitivity of this method was much higher compared to classical microcosm studies. The 13C-microcosm approach allowed the determination of first-order rate constants for 13C-labeled benzene, naphthalene, or acenaphthene even in cases when degradation activities were only small. The plausibility of the third method was checked by comparing 13C-microcosm-derived rates to field-derived rates of the first approach. Further advantage of the use of 13C-labels in microcosms is that novel metabolites can be linked more easily to specific mother compounds even in complex systems. This was achieved using alluvial sediments where 13C-acenaphthyl methylsuccinate was identified as transformation product of the anaerobic degradation of acenaphthene.

Anaerobic degradation of p-xylene by a sulfate-reducing enrichment culture

A strictly anaerobic enrichment culture was obtained with p-xylene as organic substrate and sulfate as electron acceptor from an aquifer at a former gasworks plant contaminated with aromatic hydrocarbons. p-Xylene was completely oxidized to CO2. The enrichment culture depended on Fe(II) in the medium as a scavenger of the produced sulfide. 4-Methylbenzylsuccinic acid and 4-methylphenylitaconic acid were identified in supernatants of cultures indicating that degradation of p-xylene was initiated by fumarate addition to one of the methyl groups. Therefore, p-xylene degradation probably proceeds analogously to toluene degradation by Thaueraaromatica or anaerobic degradation pathways for o- and m-xylene.

Benzene dynamics and biodegradation in alluvial aquifers affected by river fluctuations

The spatial distribution and temporal dynamics of a benzene plume in an alluvial aquifer strongly affected by river fluctuations was studied. Benzene concentrations, aquifer geochemistry datasets, past river morphology, and benzene degradation rates estimated in situ using stable carbon isotope enrichment were analyzed in concert with aquifer heterogeneity and river fluctuations. Geochemistry data demonstrated that benzene biodegradation was on-going under sulfate reducing conditions. Long-term monitoring of hydraulic heads and characterization of the alluvial aquifer formed the basis of a detailed modeled image of aquifer heterogeneity. Hydraulic conductivity was found to strongly correlate with benzene degradation, indicating that low hydraulic conductivity areas are capable of sustaining benzene anaerobic biodegradation provided the electron acceptor (SO4 (2-) ) does not become rate limiting. Modeling results demonstrated that the groundwater flux direction is reversed on annual basis when the river level rises up to 2 m, thereby forcing the infiltration of oxygenated surface water into the aquifer. The mobilization state of metal trace elements such as Zn, Cd, and As present in the aquifer predominantly depended on the strong potential gradient within the plume. However, infiltration of oxygenated water was found to trigger a change from strongly reducing to oxic conditions near the river, causing mobilization of previously immobile metal species and vice versa. MNA appears to be an appropriate remediation strategy in this type of dynamic environment provided that aquifer characterization and targeted monitoring of redox conditions are adequate and electron acceptors remain available until concentrations of toxic compounds reduce to acceptable levels.

Grundwassergefährdungspotenzial von Additiven in Wärmeträgerflüssigkeiten aus Erdwärmesonden

ZusammenfassungMit der steigenden Anzahl installierter Erdwärmesonden erhöht sich das Risiko einer Leckage bei welcher Wärmeträgerflüssigkeiten ins Grundwasser gelangen und eine lokale Verschmutzung verursachen können. In dieser Studie wurde das Grundwassergefährdungspotenzial von Additiven abgeschätzt, die in den Wärmeträgerflüssigkeiten enthalten sind. Um dies zu erreichen, war gesichertes Wissen über die chemische Identität der Additive sowie ihre Einsatzhäufigkeit und -konzentration notwendig. Eine Datenerhebung bei Herstellern und Vertreibern ergab 35 verschiedene Additive, die sich gemäß ihrer Funktion in Korrosionsinhibitoren, Alkalien, Farbstoffe, Lösungsmittel, Duftstoffe, Antischaummittel und Tenside gliedern. Das Grundwassergefährdungspotenzial der Additive wurde basierend auf ihrer Einsatzhäufigkeit und ihres Umweltverhaltens (Mobilität, Abbaubarkeit und aquatische Toxizität) abgeschätzt. Als potenziell problematisch für Grundwassersysteme müssen die Korrosionsinhibitoren Benzotriazol, Tolyltriazol, 2-Ethylhexanoat, Benzoat und Sebacat angesehen werden.AbstractThe number of installed vertical borehole heat exchangers is continuing to grow continuously which increases the risk of a local groundwater contamination with heat exchanger fluids due to a system leakage. This study evaluates the groundwater pollution potential of organic additives applied in heat exchanger fluids. In the first step of the project a data survey was carried out to collect detailed information about the identity, frequency of use and concentrations of the applied additives. In total, 35 different additives were disclosed by producers and/or distributors of heat exchanger fluids. The additives were classified as corrosion inhibitors, alkalis, dyes, solvents, scentsflavours, defoamers, and surfactants. The groundwater pollution potential of the additives was assessed based on their frequency of usage in the declared fluids and on their environmental fate (mobility, persistence and aquatic toxicity). Based on our analysis, the corrosion inhibitors benzotriazole, tolyltriazole, 2-ethylhexanoate, benzoate and sebacate exhibit a potentially exhibit a significant risk for groundwater contamination.