Anaerobic Enrichment Cultures Research Articles

Compound-specific isotope analysis as a tool to characterize biodegradation of ethylbenzene.

This study applied one- and two-dimensional compound-specific isotope analysis (CSIA) for the elements carbon and hydrogen to assess different means of microbial ethylbenzene activation. Cultures incubated under nitrate-reducing conditions showed significant carbon and highly pronounced hydrogen isotope fractionation of comparable magnitudes, leading to nearly identical slopes in dual-isotope plots. The results imply that Georgfuchsia toluolica G5G6 and an enrichment culture dominated by an Azoarcus species activate ethylbenzene by anaerobic hydroxylation catalyzed by ethylbenzene dehydrogenase, similar to Aromatoleum aromaticum EbN1. The isotope enrichment pattern in dual plots from two strictly anaerobic enrichment cultures differed considerably from those for benzylic hydroxylation, indicating an alternative anaerobic activation step, most likely fumarate addition. Large hydrogen fractionation was quantified using a recently developed Rayleigh-based approach considering hydrogen atoms at reactive sites. Data from nine investigated microbial cultures clearly suggest that two-dimensional CSIA in combination with the magnitude of hydrogen isotope fractionation is a valuable tool to distinguish ethylbenzene degradation and may be of practical use for monitoring natural or technological remediation processes at field sites.

Enriching for microbial reductive dechlorination of polychlorinated dibenzo-p-dioxins and dibenzofurans

Anaerobic enrichment cultures derived from contaminated Kymijoki River sediments dechlorinated 1,2,3,4-tetrachlorodibenzofuran (1,2,3,4-tetra-CDF), octachlorodibenzofuran (octa-CDF) and 1,2,3,4-tetrachlorodibenzo-p-dioxin (1,2,3,4-tetra-CDD). 1,2,3,4-tetra-CDF was dechlorinated via 1,2,3-, 2,3,4-, and 1,3,4/1,2,4-tri-CDFs to 1,3-, 2,3-, and 2,4-di-CDFs and finally to 4-mono-CDF. The dechlorination rate of 1,2,3,4-tetra-CDF was generally slower than that of 1,2,3,4-tetra-CDD. The rate and extent of 1,2,3,4-tetra-CDD dechlorination was enhanced by addition of pentachloronitrobenzene (PCNB) as a co-substrate. Dechlorination of spiked octa-CDF was observed with the production of hepta-, hexa-, penta- and tetra-CDFs over 6 months. Two major phylotypes of the Chloroflexi community showed an increase, one of which was identical to the Dehalococcoides mccartyi Pinellas subgroup. A set of twelve putative reductive dehalogenase (rdh) genes increased in abundance with addition of 1,2,3,4-tetra-CDF, 1,2,3,4-tetra-CDD and/or PCNB. This information will aid in understanding how indigenous microbial communities impact the fate of PCDFs and in developing strategies for bioremediation of PCDD/F contaminated sediments.

Adaptation of soil microbes during establishment of microbial fuel cell consortium fed with lactate

We report the development of microbial populations and changes in their electrochemical production during a 2-month study of a two-chamber microbial fuel cell (MFC). The original inoculum was taken from anaerobic enrichment cultures with soil as the inoculum, and lactate as the exogenous electron donor. Power density (PD), coulombic production (CP), and coulombic efficiency (CE) increased rapidly, reaching high values (320 mW m(-3), 65 Q, and 12.5%, respectively) in 12-16 days. Under these conditions, several major microbial taxa dominated the anode population. The medium solution in the cathode chamber decreased with aeration, resulting in a decrease in PD to 55 mW m(-3) at day 20. Refilling the cathode chamber around day 30 resulted in restoration of the PD, CP and CE to values equal to or greater than those previously observed. However, after the change in conditions, a marked change in community structure was observed, and high levels of acetate were seen in the anode chamber of the fuel cell for the first time. At day 35, a series of lactate concentrations were used, beginning with low levels and increasing to the 20 mM level originally used (day 46), the PD decreased but was stable at 150 mW m(-3) and the acetate concentration in the anode stabilized at about 35 mM. Under these conditions, new major population structures, which were closely related to Propionibacterium, Clostridium, and uncultured bacteria, were observed in the anode. These results suggested that the flexibility of community structure was important for sustainable electricity production.

A defined growth medium with very low background carbon for culturingClostridium thermocellum

A growth medium was developed for cultivation of Clostridium thermocellum ATCC 27405 in which "background" carbon present in buffers, reducing agents, chelating agents, and growth factors was a small fraction of the carbon present in the primary growth substrate. Background carbon was 1.6% of primary substrate carbon in the low-carbon (LC) medium, whereas it accounts for at least 40% in previously reported media. Fermentation of cellulose in LC medium was quite similar to Medium for Thermophilic Clostridia (MTC), a commonly used growth medium that contains background carbon at 88% of primary substrate carbon. Of particular note, we found that the organism can readily be cultivated by eliminating some components, lowering the concentrations of others, and employing a tenfold lower concentration of reducing agent. As such, we were able to reduce the amount of background carbon 55-fold compared to MTC medium while reaching the same cell biomass concentration. The final mass ratios of the products acetate:ethanol:formate were 5:3.9:1 for MTC and 4.1:1.5:1 for LC medium. LC medium is expected to facilitate metabolic studies involving identification and quantification of extracellular metabolites. In addition, this medium is expected to be useful in studies of cellulose utilization by anaerobic enrichment cultures obtained from environmental inocula, and in particular to diminish complications arising from metabolism of carbon-containing compounds other than cellulose. Finally, LC medium provides a starting point for industrial growth media development.

Desulfuribacillus alkaliarsenatis gen. nov. sp. nov., a deep-lineage, obligately anaerobic, dissimilatory sulfur and arsenate-reducing, haloalkaliphilic representative of the order Bacillales from soda lakes

An anaerobic enrichment culture inoculated with a sample of sediments from soda lakes of the Kulunda Steppe with elemental sulfur as electron acceptor and formate as electron donor at pH 10 and moderate salinity inoculated with sediments from soda lakes in Kulunda Steppe (Altai, Russia) resulted in the domination of a Gram-positive, spore-forming bacterium strain AHT28. The isolate is an obligate anaerobe capable of respiratory growth using elemental sulfur, thiosulfate (incomplete reduction) and arsenate as electron acceptor with H2, formate, pyruvate and lactate as electron donor. Growth was possible within a pH range from 9 to 10.5 (optimum at pH 10) and a salt concentration at pH 10 from 0.2 to 2 M total Na+ (optimum at 0.6 M). According to the phylogenetic analysis, strain AHT28 represents a deep independent lineage within the order Bacillales with a maximum of 90 % 16S rRNA gene similarity to its closest cultured representatives. On the basis of its distinct phenotype and phylogeny, the novel haloalkaliphilic anaerobe is suggested as a new genus and species, Desulfuribacillus alkaliarsenatis (type strain AHT28T = DSM24608T = UNIQEM U855T).Electronic supplementary materialThe online version of this article (doi:10.1007/s00792-012-0459-7) contains supplementary material, which is available to authorized users.

Degradation of Triclosan under Aerobic, Anoxic, and Anaerobic Conditions

Triclosan (2, 4, 4'-trichloro-2'-hydroxyl diphenyl ether) is a broad-spectrum antimicrobial agent present in a number of house hold consumables. Aerobic and anaerobic enrichment cultures tolerating triclosan were developed and 77 bacterial strains tolerating triclosan at different levels were isolated from different inoculum sources. Biodegradation of triclosan under aerobic, anoxic (denitrifying and sulphate reducing conditions), and anaerobic conditions was studied in batch cultures with isolated pure strains and enrichment consortium developed. Under aerobic conditions, the isolated strains tolerated triclosan up to 1 g/L and degraded the compound in inorganic-mineral-broth and agar media. At 10 mg/L level triclosan, 95 ± 1.2% was degraded in 5 days, producing phenol, catechol and 2, 4-dichlorophenol as the degradation products. The strains were able to metabolize triclosan and its degradation products in the presence of monooxygenase inhibitor 1-pentyne. Under anoxic/anaerobic conditions highest degradation (87%) was observed in methanogenic system with acetate as co-substrate and phenol, catechol, and 2, 4-dichlorophenol were among the products. Three of the isolated strains tolerating 1 g/L triclosan were identified as Pseudomonas sp. (BDC 1, 2, and 3).

メタン生成嫌気ベンゼン分解集積培養系に及ぼす有機酸と温度の影響

メタン生成嫌気ベンゼン分解集積培養系に及ぼす有機酸と温度の影響

Use of γ-hexachlorocyclohexane as a terminal electron acceptor by an anaerobic enrichment culture

The use of γ-hexachlorocyclohexane (HCH) as a terminal electron acceptor via organohalide respiration was demonstrated for the first time with an enrichment culture grown in a sulfate-free HEPES-buffered anaerobic mineral salts medium. The enrichment culture was initially developed with soil and groundwater from an industrial site contaminated with HCH isomers, chlorinated benzenes, and chlorinated ethenes. When hydrogen served as the electron donor, 79–90% of the electron equivalents from hydrogen were used by the enrichment culture for reductive dechlorination of the γ-HCH, which was provided at a saturation concentration of approximately 10mg/L. Benzene and chlorobenzene were the only volatile transformation products detected, accounting for 25% and 75% of the γ-HCH consumed (on a molar basis), respectively. The enrichment culture remained active with only hydrogen as the electron donor and γ-HCH as the electron acceptor through several transfers to fresh mineral salts medium for more than one year. Addition of vancomycin to the culture significantly slowed the rate of γ-HCH dechlorination, suggesting that a Gram-positive organism is responsible for the reduction of γ-HCH. Analysis of the γ-HCH dechlorinating enrichment culture did not detect any known chlororespiring genera, including Dehalobacter. In bicarbonate-buffered medium, reductive dechlorination of γ-HCH was accompanied by significant levels of acetogenesis as well as methanogenesis.

Biogenic hydrogen and methane production from Chlorella vulgaris and Dunaliella tertiolecta biomass

BackgroundMicroalgae are a promising feedstock for biofuel and bioenergy production due to their high photosynthetic efficiencies, high growth rates and no need for external organic carbon supply. In this study, utilization of Chlorella vulgaris (a fresh water microalga) and Dunaliella tertiolecta (a marine microalga) biomass was tested as a feedstock for anaerobic H2 and CH4 production.ResultsAnaerobic serum bottle assays were conducted at 37°C with enrichment cultures derived from municipal anaerobic digester sludge. Low levels of H2 were produced by anaerobic enrichment cultures, but H2 was subsequently consumed even in the presence of 2-bromoethanesulfonic acid, an inhibitor of methanogens. Without inoculation, algal biomass still produced H2 due to the activities of satellite bacteria associated with algal cultures. CH4 was produced from both types of biomass with anaerobic enrichments. Polymerase chain reaction-denaturing gradient gel electrophoresis profiling indicated the presence of H2-producing and H2-consuming bacteria in the anaerobic enrichment cultures and the presence of H2-producing bacteria among the satellite bacteria in both sources of algal biomass.ConclusionsH2 production by the satellite bacteria was comparable from D. tertiolecta (12.6 ml H2/g volatile solids (VS)) and from C. vulgaris (10.8 ml H2/g VS), whereas CH4 production was significantly higher from C. vulgaris (286 ml/g VS) than from D. tertiolecta (24 ml/g VS). The high salinity of the D. tertiolecta slurry, prohibitive to methanogens, was the probable reason for lower CH4 production.

Arsenic precipitation by an anaerobic arsenic-respiring bacterial strain isolated from the polluted sediments of Orbetello Lagoon, Italy

To isolate and characterize an anaerobic bacterial strain from the deeper polluted lagoon sediment able to use as electron acceptors [As(V)] and sulfate (SO4(2-)), using lactate as an electron donor. Methods for isolation from polluted lagoon sediments included anaerobic enrichment cultures in the presence of As(V) and SO4(2-). Reduction of As(V) to As(III) was observed during the growth of the bacterial strain, and the final concentration of As(III) was lower than the initial As(V) one, suggesting the immobilization of As(III) in the yellow precipitate. The precipitate was identified by energy dispersive spectroscopy X-ray as arsenic sulfide. Scanning electron microscopy (SEM) revealed rod-shaped bacterial cells embedded in the precipitate, where net-like formations strictly related to the bacterial cells were visible. The surface of the precipitate showed the adhesion of bacterial cells, forming clusters. Transmission electron microscopy (TEM) also highlighted precipitates inside the bacterial cells and on their surface. Following 16S rRNA sequencing, the bacterial strain 063 was assigned to the genus Desulfosporosinus. This study reports, for the first time, the isolation from the polluted lagoon sediments of a strain capable of respiring and using As(V) and SO4(2-) as electron acceptors with lactate as the sole carbon and energy source with the formation of an arsenic sulfide precipitate. The identification of these properties provides novel insight into the possible use of the anaerobic strain in bioremediation processes and also adds to the knowledge on the biogeochemical cycling of arsenic.

Anaerobic phototrophic nitrite oxidation by Thiocapsa sp. strain KS1 and Rhodopseudomonas sp. strain LQ17

In anaerobic enrichment cultures for phototrophic nitrite-oxidizing bacteria from different freshwater sites, two different cell types, i.e. non-motile cocci and motile, rod-shaped bacteria, always outnumbered all other bacteria. Most-probable-number (MPN) dilution series with samples from two freshwater sites yielded only low numbers (<or=3x10(3) cm(-3)) of phototrophic nitrite oxidizers. Slightly higher numbers (about 10(4) cm(-3)) were found in activated sewage sludge. Anaerobic phototrophic oxidation of nitrite was studied with two different isolates, the phototrophic sulfur bacterium strain KS1 and the purple nonsulfur bacterium strain LQ17, both of which were isolated from activated sludge collected from the municipal sewage treatment plant in Konstanz, Germany. Strain KS1 converted 1 mM nitrite stoichiometrically to nitrate with concomitant formation of cell matter within 2-3 days, whereas strain LQ17 oxidized only up to 60 % of the given nitrite to nitrate within several months with the concomitant formation of cell biomass. Nitrite oxidation to nitrate was strictly light-dependent and required the presence of molybdenum in the medium. Nitrite was oxidized in both the presence and absence of oxygen. Nitrite inhibited growth at concentrations higher than 2 mM. Hydroxylamine and hydrazine were found to be toxic to the phototrophs in the range 5-50 muM and did not stimulate phototrophic growth. Based on morphology, substrate-utilization pattern, in vivo absorption spectra, and 16S rRNA gene sequence similarity, strain KS1 was assigned to the genus Thiocapsa and strain LQ17 to the genus Rhodopseudomonas. Also, Thiocapsa roseopersicina strains DSM 217 and DSM 221 were found to oxidize nitrite to nitrate with concomitant growth. We conclude that the ability to use nitrite phototrophically as electron donor is widespread in nature, but low MPN counts indicate that its contribution to nitrite oxidation in the studied habitats is rather limited.

Desulfurispira natronophila gen. nov. sp. nov.: an obligately anaerobic dissimilatory sulfur-reducing bacterium from soda lakes

Anaerobic enrichment cultures with elemental sulfur as electron acceptor and either acetate or propionate as electron donor and carbon source at pH 10 and moderate salinity inoculated with sediments from soda lakes in Kulunda Steppe (Altai, Russia) resulted in the isolation of two novel members of the bacterial phylum Chrysiogenetes. The isolates, AHT11 and AHT19, represent the first specialized obligate anaerobic dissimilatory sulfur respirers from soda lakes. They use either elemental sulfur/polysulfide or arsenate as electron acceptor and a few simple organic compounds as electron donor and carbon source. Elemental sulfur is reduced to sulfide through intermediate polysulfide, while arsenate is reduced to arsenite. The bacteria belong to the obligate haloalkaliphiles, with a pH growth optimum from 10 to 10.2 and a salt range from 0.2 to 3.0 M Na+ (optimum 0.4–0.6 M). According to the phylogenetic analysis, the two strains were close to each other, but distinct from the nearest relative, the haloalkaliphilic sulfur-reducing bacterium Desulfurispirillum alkaliphilum, which was isolated from a bioreactor. On the basis of distinct phenotype and phylogeny, the soda lake isolates are proposed as a new genus and species, Desulfurispira natronophila (type strain AHT11T = DSM22071T = UNIQEM U758T).Electronic supplementary materialThe online version of this article (doi:10.1007/s00792-010-0314-7) contains supplementary material, which is available to authorized users.

Biodegradation of naphthalene, phenanthrene, and pyrene under anaerobic conditions

The biodegradation of naphthalene, phenanthrene, and pyrene were studied in batch reactors using an anaerobic acetate-fed enrichment culture developed from the Greater Peoria Sanitary District Wastewater Treatment Facility. Baseline kinetic experiments showed that maximum specific substrate utilization for the acetate-fed enrichment culture was 2.72 mg acetate/mg cell/day and the endogenous decay coefficient was 0.043 day−1. Maximum specific substrate utilization rates in mg substrate/mg cell/day for naphthalene, phenanthrene and pyrene were 0.57, 0.009 and 0.007, respectively. The half saturation constant KS in mg /L were 0.075, 0.01 and less than 0.01, respectively. The endogenous decay coefficient (in units of day−1) in the presence of naphthalene, phenanthrene and pyrene were 0.06, 0.07 and 0.04, respectively. Results from this study indicated that naphthalene and phenanthrene were more easily biodegradable than pyrene. These results suggest that technologies using anaerobic biodegradation using a relatively simple enrichment culture from a wastewater treatment plant may be successful in treating PAH contaminated wastes.

Sulfate-Mediated Bacterial Population Shift in a Hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-Degrading Anaerobic Enrichment Culture

ABSTRACT The effects of sulfate on the population dynamics of an anaerobic hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX)-degrading consortium were studied using terminal restriction fragment length polymorphism (T-RFLP) analysis. One hundred percent of the initial RDX was degraded in the sulfate-amended culture within 3 days of incubation. In the sulfate-unamended cultures, 35% of the initial RDX remained after 3 days and 8% after 7 days of incubation. Based on the T-RFLP distribution of the community 16S rDNA genes, the microcosm consisted predominantly of two organisms, a Geobacter sp. (78%) and an Acetobacterium sp. (14%). However, in the presence of sulfate, both species decreased to less than 3% of the total population within 3 days and an unclassified Clostridiaceae became the dominant organism at 40% the total fragment distribution. This indicated the explosive-degrading consortium had greater diversity than initially perceived and rapidly adapted to a readily available electron acceptor, which in turn stimulated RDX degradation.

Characterization of a Dehalobacter Coculture That Dechlorinates 1,2-Dichloroethane to Ethene and Identification of the Putative Reductive Dehalogenase Gene

Dehalobacter and "Dehalococcoides" spp. were previously shown to be involved in the biotransformation of 1,1,2-trichloroethane (1,1,2-TCA) and 1,2-dichloroethane (1,2-DCA) to ethene in a mixed anaerobic enrichment culture. Here we report the further enrichment and characterization of a Dehalobacter sp. from this mixed culture in coculture with an Acetobacterium sp. Through a series of serial transfers and dilutions with acetate, H(2), and 1,2-DCA, a stable coculture of Acetobacterium and Dehalobacter spp. was obtained, where Dehalobacter grew during dechlorination. The isolated Acetobacterium strain did not dechlorinate 1,2-DCA. Quantitative PCR with specific primers showed that Dehalobacter cells did not grow in the absence of a chlorinated electron acceptor and that the growth yield with 1,2-DCA was 6.9 (+/-0.7) x 10(7) 16S rRNA gene copies/mumol 1,2-DCA degraded. PCR with degenerate primers targeting reductive dehalogenase genes detected three distinct Dehalobacter/Desulfitobacterium-type sequences in the mixed-parent culture, but only one of these was present in the 1,2-DCA-H(2) coculture. Reverse transcriptase PCR revealed the transcription of this dehalogenase gene specifically during the dechlorination of 1,2-DCA. The 1,2-DCA-H(2) coculture could dechlorinate 1,2-DCA but not 1,1,2-TCA, nor could it dechlorinate chlorinated ethenes. As a collective, the genus Dehalobacter has been show to dechlorinate many diverse compounds, but individual species seem to each have a narrow substrate range.

Ecophysiology of “ Halarsenatibacter silvermanii ” Strain SLAS-1 T , gen. nov., sp. nov., a Facultative Chemoautotrophic Arsenate Respirer from Salt-Saturated Searles Lake, California

Searles Lake occupies a closed basin harboring salt-saturated, alkaline brines that have exceptionally high concentrations of arsenic oxyanions. Strain SLAS-1(T) was previously isolated from Searles Lake (R. S. Oremland, T. R. Kulp, J. Switzer Blum, S. E. Hoeft, S. Baesman, L. G. Miller, and J. F. Stolz, Science 308:1305-1308, 2005). We now describe this extremophile with regard to its substrate affinities, its unusual mode of motility, sequenced arrABD gene cluster, cell envelope lipids, and its phylogenetic alignment within the order Halanaerobacteriales, assigning it the name "Halarsenatibacter silvermanii" strain SLAS-1(T). We also report on the substrate dynamics of an anaerobic enrichment culture obtained from Searles Lake that grows under conditions of salt saturation and whose members include a novel sulfate reducer of the order Desulfovibriales, the archaeon Halorhabdus utahensis, as well as a close homolog of strain SLAS-1(T).

Heterotrophic denitrification at extremely high salt and pH by haloalkaliphilic Gammaproteobacteria from hypersaline soda lakes

In this paper we describe denitrification at extremely high salt and pH in sediments from hypersaline alkaline soda lakes and soda soils. Experiments with sediment slurries demonstrated the presence of acetate-utilizing denitrifying populations active at in situ conditions. Anaerobic enrichment cultures at pH 10 and 4 M total Na+ with acetate as electron donor and nitrate, nitrite and N2O as electron acceptors resulted in the dominance of Gammaproteobacteria belonging to the genus Halomonas. Both mixed and pure culture studies identified nitrite and N2O reduction as rate-limiting steps in the denitrification process at extremely haloalkaline conditions.Electronic supplementary materialThe online version of this article (doi:10.1007/s00792-008-0166-6) contains supplementary material, which is available to authorized users.

Sulfidogenesis under extremely haloalkaline conditions by Desulfonatronospira thiodismutans gen. nov., sp. nov., and Desulfonatronospira delicata sp. nov. - a novel lineage of Deltaproteobacteria from hypersaline soda lakes

High rates of sulfidogenesis were observed in sediments from hypersaline soda lakes. Anaerobic enrichment cultures at 2 M Na(+) and pH 10 inoculated with sediment samples from these lakes produced sulfide most actively with sulfite and thiosulfate as electron acceptors, and resulted in the isolation of three pure cultures of extremely natronophilic sulfidogenic bacteria. Strain ASO3-1 was isolated using sulfite as a sole substrate, strain AHT 8 with thiosulfate and formate, and strain AHT 6 with thiosulfate and acetate. All strains grew in a mineral soda-based medium by dismutation of either sulfite or thiosulfate, as well as with sulfite, thiosulfate and sulfate as acceptors, and H(2) and simple organic compounds as electron donors. The acetyl-CoA pathway was identified as the pathway for inorganic carbon assimilation by strain ASO3-1. All strains were obligately alkaliphilic, with an optimum at pH 9.5-10, and grew in soda brines containing 1-4 M total Na(+) (optimum at 1.0-2.0 M). The cells accumulated high amounts of the organic osmolyte glycine betaine. They formed a new lineage within the family Desulfohalobiaceae (Deltaproteobacteria), for which the name Desulfonatronospira gen. nov. is proposed. Strains ASO3-1(T) and AHT 8 from Kulunda Steppe formed Desulfonatronospira thiodismutans sp. nov., and strain AHT 6(T) from Wadi al Natrun is suggested as Desulfonatronospira delicata sp. nov.

Microbial Dehalogenation of Trichlorinated Dibenzo-p-dioxins by a Dehalococcoides-Containing Mixed Culture Is Coupled to Carbon Isotope Fractionation

An anaerobic enrichment culture reductively dehalogenated 1,2,4- and 1,2,3-trichlorodibenzo-p-dioxin (TrCDD) almost exclusively at peripheral positions forming the main products 1,3-dichloro-(DiCDD) and 2-monochlorodibenzo-p-dioxin (MCDD) from 1,2,4-TrCDD and 2,3-DiCDD from 1,2,3-TrCDD. Dehalococcoides was monitored in the mixed culture by quantitative real-time PCR. A yield of 2.5 x 10(8) to 2.75 x 10(8) copies of 16S rRNA genes per micromole of chloride released suggested growth by dehalorespiration with dibenzo-p-dioxins. For the analysis of carbon isotope fractionation, the dioxin congeners were isolated by solid-phase microextraction (SPME) from the headspace of the cultures. The delta13C composition of 1,2,4-TrCDD did not change remarkably during the course of reductive dehalogenation; however, the intermediate 1,3-DiCDD became enriched, and the final product 2-MCDD significantly depleted in 13C with a discrimination of 2.5-3.6 per thousand between 1,3-DiCDD and 2-MCDD. 1,2,3-TrCDD and its main product 2,3-DiCDD became slightly enriched in 13C, whereas the formed low concentrations of 2-MCDD were depleted in 13C by 5.5-4.8 per thousand. This study demonstrates carbon isotope fractionation during sequential reductive dehalogenation of chlorinated dibenzo-p-dioxins, whereby isotope fractionation upon dehalogenation of the intermediate was substantial. This can provide a basis for the development of a new method to monitor the fate of dioxins in the environment using compound specific stable isotope analyses.

Investigation of Reductive Dechlorination Supported by Natural Organic Carbon

AbstractBecause remediation timeframes using monitored natural attenuation may span decades or even centuries at chlorinated solvent sites, new approaches are needed to assess the long‐term sustainability of reductive dechlorination in ground water systems. In this study, extraction procedures were used to investigate the mass of indigenous organic carbon in aquifer sediment, and experiments were conducted to determine if the extracted carbon could support reductive dechlorination of chloroethenes. Aquifer sediment cores were collected from a site without an anthropogenic source of organic carbon where organic carbon varied from 0.02% to 0.12%. Single extraction results showed that 1% to 28% of sediment‐associated organic carbon and 2% to 36% of the soft carbon were removed depending on nature and concentration of the extracting solution (Nanopure water; 0.1%, 0.5%, and 1.0% sodium pyrophosphate; and 0.5 N sodium hydroxide). Soft carbon is defined as organic carbon oxidized with potassium persulfate and is assumed to serve as a source of biodegradable carbon within the aquifer. Biodegradability studies demonstrated that 20% to 40% of extracted organic carbon was biodegraded aerobically and anaerobically by soil microorganisms in relatively brief tests (45 d). A five‐step extraction procedure consisting of 0.1% pyrophosphate and base solutions was investigated to quantify bioavailable organic carbon. Using the extracted carbon as the sole electron donor source, tetrachloroethene was transformed to cis‐1,2‐dichloroethene and vinyl chloride in anaerobic enrichment culture experiments. Hydrogen gas was produced at levels necessary to sustain reductive dechlorination (&gt;1 nM).