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Anaerobic oxidation of [1,2- 14 C]dichloroethene to 14 CO 2 under Mn(IV)-reducing conditions was demonstrated. The results indicate that oxidative degradation of partially chlorinated solvents like dichloroethene can be significant even under anoxic conditions and demonstrate the potential importance of Mn(IV) reduction for remediation of chlorinated groundwater contaminants.
Life in the presence of high salt concentrations is compatible with life in the absence of oxygen. Halophilic and halotolerant anaerobic prokaryotes are found both in the archaeal and in the bacterial domain, and they display a great metabolic diversity. Many of the representatives of the Halobacteriales (Archaea), which are generally considered aerobes, have the potential of anaerobic growth. Some can use alternative electron acceptors such as nitrate, fumarate, dimethylsulfoxide or trimethylamine-N-oxide Halobacterium salinarum can also grow fermentatively on L-arginine, and bacteriorhodopsin-containing cells may even grow anaerobically, energized by light. Obligatory anaerobic halophilic methanogenic Archaea also exist. The bacterial domain contains many anaerobic halophiles, including sulfate reducers. There is also a group of specialized obligatory anaerobic Bacteria, phylogenetically clustering in the low G+C branch of the Firmicutes. Most representatives of this group (order Haloanaerobiales , families Haloanaerobiaceae and Halobacteroidaceae ) are fermentative, using a variety of carbohydrates and amino acids. One species combines the potential for anaerobic growth at high salt concentrations with a preference for high temperatures. Others are homoacetogens; Acetohalobium arabaticum can grow anaerobically as a chemolithotroph, producing acetate from hydrogen and CO 2 . The Haloanaerobiales accumulate high concentrations of K + and Cl - in their cytoplasm, thereby showing a strategy of salt adaptation similar to that used by the Halobacteriales . Recently a new representative of the Haloanaerobiales was isolated from bottom sediments of the Dead Sea (strain DSSe1), which grows anaerobically by oxidation of glycerol to acetate and CO 2 while reducing selenate to selenite and elementary selenium. Other electron acceptors supporting anaerobic growth of this strain are nitrate and trimethylamine- N -oxide. The versatility of life at high salt concentrations with respect to the variety of substrates used, the types of dissimilatory metabolism, and the diversity of potential electron acceptors has important implications for the potential for life in hostile environments lacking oxygen and high in salt, implications that may also be relevant to astrobiology.
Anaerobic digestion has been suggested as an intervention to attenuate antibiotic resistance genes (ARGs) in livestock manure but supporting data have typically been collected at laboratory scale. Few studies have quantified ARG fate during full-scale digestion of livestock manure. We sampled untreated manure and digestate from seven full-scale mesophilic dairy manure digesters to assess ARG fate through each system. Samples were collected biweekly from December through August (i.e., winter, spring, and summer; n = 235 total) and analyzed by quantitative polymerase chain reaction for intI1 , erm (B), sul1 , tet (A), and tet (W). Concentrations of intI1 , sul1 , and tet (A) decreased during anaerobic digestion, but their removal was less extensive than expected based on previous laboratory studies. Removal for intI1 during anaerobic digestion equaled 0.28 ± 0.03 log 10 units (mean ± SE), equivalent to only 48% removal and notable given intI1 ’s role in horizontal gene transfer and multiple resistance. Furthermore, tet (W) concentrations were unchanged during anaerobic digestion ( p > 0.05), and erm (B) concentrations increased by 0.52 ± 0.03 log 10 units (3.3-fold), which is important given erythromycin's status as a critically important antibiotic for human medicine. Seasonal log 10 changes in intI1 , sul1 , and tet (A) concentrations were ≥50% of corresponding log 10 removals by anaerobic digestion, and variation in ARG and intI1 concentrations among digesters was quantitatively comparable to anaerobic digestion effects. These results suggest that mesophilic anaerobic digestion may be limited as an intervention for ARGs in livestock manure and emphasize the need for multiple farm-level interventions to attenuate antibiotic resistance.
To test the effects of altered hydrology on organic soil decomposition, we investigated CO 2 and CH 4 production potential of rich-fen peat (mean surface pH = 6.3) collected from a field water table manipulation experiment including control, raised and lowered water table treatments. Mean anaerobic CO 2 production potential at 10 cm depth (14.1 ± 0.9 μmol C g −1 d −1 ) was as high as aerobic CO 2 production potential (10.6 ± 1.5 μmol C g −1 d −1 ), while CH4 production was low (mean of 7.8 ± 1.5 nmol C g −1 d −1 ). Denitrification enzyme activity indicated a very high denitrification potential (197 ± 23 μg N g −1 d −1 ), but net NO - 3 reduction suggested this was a relatively minor pathway for anaerobic CO 2 production. Abundances of denitrifier genes ( nirK and nosZ ) did not change across water table treatments. SO 2- 4 reduction also did not appear to be an important pathway for anaerobic CO 2 production. The net accumulation of acetate and formate as decomposition end products in the raised water table treatment suggested that fermentation was a significant pathway for carbon mineralization, even in the presence of NO - 3 . Dissolved organic carbon (DOC) concentrations were the strongest predictors of potential anaerobic and aerobic CO 2 production. Across all water table treatments, the CO 2 :CH 4 ratio increased with initial DOC leachate concentrations. While the field water table treatment did not have a significant effect on mean CO 2 or CH 4 production potential, the CO 2 :CH 4 ratio was highest in shallow peat incubations from the drained treatment. These data suggest that with continued drying or with a more variable water table, anaerobic CO 2 production may be favored over CH 4 production in this rich fen. Future research examining the potential for dissolved organic substances to facilitate anaerobic respiration, or alternative redox processes that limit the effectiveness of organic acids as substrates in anaerobic metabolism, would help explain additional uncertainty concerning carbon mineralization in this system.
The potential for anaerobic benzene oxidation in the Fe(III)-reduction zone of petroleum-contaminated aquifers was evaluated. Sediments were incubated under strict anaerobic conditions without any amendments in order to simulate in situ conditions. [ 14 C]Benzene was not oxidized to 14 CO 2 at most sites examined, which is consistent with previous studies that have found that benzene tends to persist in unamended, anaerobic aquifer materials and/or long periods of time are required in order to adapt the microbial population for benzene degradation. However, at one site located in Bemidji, MN, [ 14 C]benzene was oxidized to 14 CO 2 in unamended sediments without an apparent lag, suggesting that benzene was anaerobically degraded in situ. Benzene was not significantly oxidized in sediments from nearby Fe(III)-reducing sites nor in sediments collected from an uncontaminated background site in the same aquifer. Culturing and 16S rRNA-based molecular studies of the Bemidji aquifer demonstrated that while all sites contained similar numbers of Fe(III)-reducing microorganisms closely related to Geothrix fermentens , the site at which anaerobic benzene degradation was observed was greatly enriched with microorganisms in the family Geobacteraceae . This study provides the first data consistent with in situ anaerobic oxidation of benzene to carbon dioxide in the Fe(III)-reducing zone of a petroleum-contaminated aquifer and suggests that comparative studies on the size of the Geobacteraceae community in petroleum-contaminated aquifers might aid in the location of zones in which benzene degradation coupled to Fe(III) reduction is taking place.
Kinetic isotopic fractionation of carbon and nitrogen during RDX (hexahydro-1,3,5-trinitro-1,3,5-triazine) biodegradation was investigated with pure bacterial cultures under aerobic and anaerobic conditions. Relatively large bulk enrichments in 15 N were observed during biodegradation of RDX via anaerobic ring cleavage (ε 15 N = −12.7‰ ± 0.8‰) and anaerobic nitro reduction (ε 15 N = −9.9‰ ± 0.7‰), in comparison to smaller effects during biodegradation via aerobic denitration (ε 15 N = −2.4‰ ± 0.2‰). 13 C enrichment was negligible during aerobic RDX biodegradation (ε 13 C = −0.8‰ ± 0.5‰) but larger during anaerobic degradation (ε 13 C = −4.0‰ ± 0.8‰), with modest variability among genera. Dual-isotope ε 13 C/ε 15 N analyses indicated that the three biodegradation pathways could be distinguished isotopically from each other and from abiotic degradation mechanisms. Compared to the initial RDX bulk δ 15 N value of +9‰, δ 15 N values of the NO 2 − released from RDX ranged from −7‰ to +2‰ during aerobic biodegradation and from −42‰ to −24‰ during anaerobic biodegradation. Numerical reaction models indicated that N isotope effects of NO 2 − production were much larger than, but systematically related to, the bulk RDX N isotope effects with different bacteria. Apparent intrinsic ε 15 N-NO 2 − values were consistent with an initial denitration pathway in the aerobic experiments and more complex processes of NO 2 − formation associated with anaerobic ring cleavage. These results indicate the potential for isotopic analysis of residual RDX for the differentiation of degradation pathways and indicate that further efforts to examine the isotopic composition of potential RDX degradation products (e.g., NO x ) in the environment are warranted.
Enrichment cultures that anaerobically degraded oxalate were obtained from lake sediment inocula. From these, 5 pure cultures of anaerobic oxalate-degrading bacteria were isolated and partially characterized. The isolates were Gram-negative, non-sporeforming, non-motile, obligate anaerobes. Oxalate was required for growth and was stoichiometrically converted to formate; 14 CO 2 was also recovered when 14 C-oxalate was added. Maximal growth occurred when the oxalate concentration was 50 mM. Acetate stimulated growth in the presence of oxalate, however, 14 C-experiments indicated that acetate was only utilized for cell carbon. The isolates were either spiral-shaped or rod-shaped organisms. The first morphotype grew much more slowly than the second and exhibited 13-fold lower cell yields. These isolates represent a new strain of oxalate-degrading bacteria. The second morphotype was similar to the anaerobic oxalate-degrading bacteria previously found in rumen. This report extends the known habitats in which anaerobic oxalate-degrading organisms have been found to include aquatic sediments.
Background. Arsenic speciation in environmental samples is essential for studying toxicity, mobility and bio-transformation of As in aquatic and terrestrial environments. Although the inorganic species As(III) and As(V) have been considered dominant in soils and sediments, organisms are able to metabolize inorganic forms of arsenic into organo-arsenic compounds. Arsenosugars and methylated As compounds can be found in terrestrial organisms, but they generally occur only as minor constituents. We investigated the dynamics of arsenic species under anaerobic conditions in soils surrounding gold mining areas from Minas Gerais State, Brazil to elucidate the arsenic biogeochemical cycle and water contamination mechanisms. Methods. Surface soil samples were collected at those sites, namely Paracatu Formation, Banded Iron Formation and Riacho dos Machados Sequence, and incubated in CaCl2 2.5 mmol L-1 suspensions under anaerobic conditions for 1, 28, 56 and 112 days. After that, suspensions were centrifuged and supernatants analyzed for soluble As species by IC-ICPMS and HPLC-ICPMS. Results. Easily exchangeable As was mainly arsenite, except when reducible manganese was present. Arsenate was mainly responsible for the increase in soluble arsenic due to the reductive dissolution of either iron or manganese in samples from the Paracatu Formation and Riacho dos Machados Sequence. On the other hand, organic species of As dominated in samples from the Banded Iron Formation during anaerobic incubation. Discussion. Results are contrary to the expectation that, in anaerobic environments, As release due to the reductive dissolution of Fe is followed by As(V) reduction to As(III). The occurrence of organo-arsenic species was also found to be significant to the dynamics of soluble arsenic, mainly in soils from the Banded Iron Formation (BIF), under our experimental conditions. Conclusions. In general, As(V) and organic As were the dominant species in solution, which is surprising under anaerobic conditions in terrestrial environments. The unexpected occurrence of organic species of As was attributed to enrollment of ternary organic complexes or living organisms such as algae or cyanobacteria. Perspectives. These findings are believed to be useful for remediation strategies in mine-affected regions, as the organic As species are in general considered to be less toxic than inorganic ones and even As(V) is considered less mobile and toxic than As(III). ?? 2007 ecomed publishers (Verlagsgruppe Hu??thig Jehle Rehm GmbH).
We isolated an obligately anaerobic halophilic bacterium from the Dead Sea that grew by respiration of selenate. The isolate, designated strain DSSe-1, was a gram-negative, non-motile rod. It oxidized glycerol or glucose to acetate+CO2 with concomitant reduction of selenate to selenite plus elemental selenium. Other electron acceptors that supported anaerobic growth on glycerol were nitrate and trimethylamine-N-oxide; nitrite, arsenate, fumarate, dimethylsulfoxide, thiosulfate, elemental sulfur, sulfite or sulfate could not serve as electron acceptors. Growth on glycerol in the presence of nitrate occurred over a salinity range from 100 to 240 g/l, with an optimum at 210 g/l. Analysis of the 16S rRNA gene sequence suggests that strain DSSe-1 belongs to the order Halanaerobiales, an order of halophilic anaerobes with a fermentative or homoacetogenic metabolism, in which anaerobic respiratory metabolism has never been documented. The highest 16S rRNA sequence similarity (90%) was found with Acetohalobium arabaticum (X89077). On the basis of physiological properties as well as the relatively low homology of 16S rRNA from strain DSSe-1 with known genera, classification in a new genus within the order Halanaerobiales, family Halobacteroidaceae is warranted. We propose the name Selenihalanaerobacter shriftii. Type strain is strain DSSe-1 (ATCC accession number BAA-73).
The potential use of iron(III) oxide to stimulate in-situ hydrocarbon degradation in anaerobic petroleum-contaminated harbor sediments was investigated. Previous studies have indicated that Fe(III)-reducing bacteria (FeRB) can oxidize some electron donors more effectively than sulfate- reducing bacteria (SRB). In contrast to previous results in freshwater sediments, the addition of Fe(III) to marine sediments from San Diego Bay, CA did not switch the terminal electron-accepting process (TEAP) from sulfate reduction to Fe-(III) reduction. Addition of Fe(III) also did not stimulate anaerobic hydrocarbon oxidation. Exposure of the sediment to air [to reoxidize Fe(II) to Fe(III)] followed by anaerobic incubation of the sediments, resulted in Fe-(III) reduction as the TEAP, but contaminant degradation was not stimulated and in some instances was inhibited. The difference in the ability of FeRB to compete with the SRB in the different sediment treatments was related to relative population sizes. Although the addition of Fe(III) did not stimulate hydrocarbon degradation, the results presented here as well as other recent studies demonstrate that there may be significant anaerobic hydrocarbon degradation under sulfate-reducing conditions in harbor sediments.
Antibiotics are used to maintain healthy livestock and to promote weight gain in concentrated animal feed operations. Antibiotics rarely are metabolized completely by livestock and, thus, are often present in livestock waste and in waste-treatment lagoons. The introduction of antibiotics into anaerobic lagoons commonly used for swine waste treatment has the potential for negative impacts on lagoon performance, which relies on a consortium of microbes ranging from fermentative microorganisms to methanogens. To address this concern, the effects of eight common veterinary antibiotics on anaerobic activity were studied. Anaerobic microcosms, prepared from freshly collected lagoon slurries, were amended with individual antibiotics at 10 mg/L for the initial screening study and at 1, 5, and 25 mg/L for the dose-response study. Monitored metabolic indicators included hydrogen, methane, and volatile fatty acid concentrations as well as chemical oxygen demand. The selected antibiotics significantly inhibited methane production relative to unamended controls, thus indicating that antibiotics at concentrations commonly found in swine lagoons can negatively impact anaerobic metabolism. Additionally, historical antibiotic usage seems to be a potential factor in affecting methane production. Specifically, less inhibition of methane production was noted in samples taken from the lagoon with a history of multiple-antibiotic use.
The alluvial aquifer adjacent to Norman Landfill, OK, provides an excellent natural laboratory for the study of anaerobic processes impacting landfill-leachate contaminated aquifers. We collected groundwaters from a transect of seven multilevel wells ranging in depth from 1.3 to 11 m that were oriented parallel to the flow path. The center of the leachate plume was characterized by (1) high alkalinity and elevated concentrations of total dissolved organic carbon, reduced iron, and methane, and (2) negligible oxygen, nitrate, and sulfate concentrations. Methane concentrations and stable carbon isotope (δ 13 C) values suggest anaerobic methane oxidation was occurring within the plume and at its margins. Methane δ 13 C values increased from about −54‰ near the source to >−10‰ downgradient and at the plume margins. The isotopic fractionation associated with this methane oxidation was −13.6 ± 1.0‰. Methane 13 C enrichment indicated that 80−90% of the original landfill methane was oxidized over the 210-m transect. First-order rate constants ranged from 0.06 to 0.23 per year, and oxidation rates ranged from 18 to 230 μM/y. Overall, hydrochemical data suggest that a sulfate reducer-methanogen consortium may mediate this methane oxidation. These results demonstrate that natural attenuation through anaerobic methane oxidation can be an important sink for landfill methane in aquifer systems.
Benzene has often been observed to be resistant to microbial degradation under anoxic conditions. A number of recent studies, however, have demonstrated that anaerobic benzene utilization can occur. This study extends the previous reports of anaerobic benzene degradation to sediments that varied with respect to contamination input, predominant redox condition, and salinity. In spite of differences in methodology, microbial degradation of benzene was noted in slurries constructed with sediments from various geographical locations and range from aquifer sands to fine-grained estuarine muds, under methanogenic, sulfate-reducing, and iron-reducing conditions. In aquifer sediments under methanogenic conditions, benzene loss was concomitant with methane production, and microbial utilization of [ 14 C]benzene yielded 14 CO 2 and 14 CH 4 . In slurries with estuarine and aquifer sediments under sulfate-reducing conditions, the loss of sulfate in amounts consistent with the stoichiometric degradation of benzene or the conversion of [ 14 C]benzene to 14 CO 2 indicates that benzene was mineralized. Benzene loss also occurred in the presence of Fe(III) in sediments from freshwater environments. Microbial benzene utilization, however, was not observed under denitrifying conditions. These results indicate that the potential for the anaerobic degradation of benzene, which was once thought to be resistant to non-oxygenase attack, exists in a variety of aquatic sediments from widely distributed locations.
Monomethylarsonic acid (MMAV) and dimethylarsinic acid (DMAV) are extensively utilized as pesticides, introducing large quantities of arsenic into the environment. Once released into the environment, these organoarsenicals are subject to microbial reactions. Aerobic biodegradation of MMAV and DMAV has been evaluated, but little is known about their fate in anaerobic environments. The objective of this study was to evaluate the biotransformation of MMAV and DMAV in anaerobic sludge. Biologically mediated conversion occurred under methanogenic or sulfate-reducing conditions but not in the presence of nitrate. Monomethylarsonous acid (MMAIII) was consistently observed as an important metabolite of MMAV degradation, and it was recovered in molar yields ranging from 5 to 47%. The main biotransformation product identified from DMAV metabolism was MMAV, which was recovered in molar yields ranging from 8 to 65%. The metabolites indicate that reduction and demethylation are important steps in the anaerobic bioconversion of MMAV and DMAV, respectively. ?? 2006 American Chemical Society.
Iron mineral transformations occurring in hydrocarbon-contaminated sites are linked to the biodegradation of the hydrocarbons. At a hydrocarbon-contaminated site near Bemidji, Minnesota, USA, measurements of magnetic susceptibility (MS) are useful for monitoring the natural attenuation of hydrocarbons related to iron cycling. However, a transient MS, previously observed at the site, remains poorly understood and the iron mineral phases acting as reactants and products associated with this MS perturbation remain largely unknown. To address these unknowns, we acquired mineral magnetism measurements, including hysteresis loops, backfield curves, and isothermal remanent magnetizations on sediment core samples retrieved from the site and magnetite-filled mineral packets installed within the aquifer. Our data show that the core samples and magnetite packs display decreasing magnetization with time and that this loss in magnetization is accompanied by increasing bulk coercivity consistent with decreased average grain size and/or partial oxidation. Low-temperature magnetometry on all samples displayed behavior consistent with magnetite, but samples within the plume also show evidence of maghemitization. This interpretation is supported by the occurrence of shrinkage cracks on the surface of the grains imaged via scanning electron microscopy. Magnetite transformation to maghemite typically occurs under oxic conditions, here, we propose that maghemitization occurs within the anoxic portions of the plume via microbially mediated anaerobic oxidation. Mineral dissolution also occurs within the plume. Microorganisms capable of such anaerobic oxidation have been identified within other areas at the Bemidji site, but additional microbiological studies are needed to link specific anaerobic iron oxidizers with this loss of magnetization.
A two-dimensional, multispecies reactive solute transport model with sequential aerobic and anaerobic degradation processes was developed and tested. The model was used to study the field-scale solute transport and degradation processes at the Bemidji, Minnesota, crude oil spill site. The simulations included the biodegradation of volatile and nonvolatile fractions of dissolved organic carbon by aerobic processes, manganese and iron reduction, and methanogenesis. Model parameter estimates were constrained by published Monod kinetic parameters, theoretical yield estimates, and field biomass measurements. Despite the considerable uncertainty in the model parameter estimates, results of simulations reproduced the general features of the observed groundwater plume and the measured bacterial concentrations. In the simulation, 46% of the total dissolved organic carbon (TDOC) introduced into the aquifer was degraded. Aerobic degradation accounted for 40% of the TDOC degraded. Anaerobic processes accounted for the remaining 60% of degradation of TDOC: 5% by Mn reduction, 19% by Fe reduction, and 36% by methanogenesis. Thus anaerobic processes account for more than half of the removal of DOC at this site.
The potential contribution of microbial metabolism to the magnetization of sediments has only recently been recognized. In the presence of oxygen, magnetotactic bacteria can form intracellular chains of magnetite while using oxygen or nitrate as the terminal electron acceptor for metabolism 1 . The production of ultrafine-grained magnetite by magnetotactic bacteria in surficial aerobic sediments may contribute significantly to the natural remanent magnetism of sediments 2–4 . However, recent studies on iron reduction in anaerobic sediments suggested that bacteria can also generate magnetite in the absence of oxygen 5 . We report here on a sediment organism, designated GS-15, which produces copious quantities of ultrafine-grained magnetite under anaerobic conditions. GS-15 is not magnetotactic, but reduces amorphic ferric oxide to extracellular magnetite during the reduction of ferric iron as the terminal electron acceptor for organic matter oxidation. This novel metabolism may be the mechanism for the formation of ultrafine-grained magnetite in anaerobic sediments, and could account for the accumulation of magnetite in ancient iron formations and hydrocarbon deposits.
Collectively, marine sediments comprise the largest reservoir of methane on Earth. The flux of methane from the sea bed to the overlying water column is mitigated by the sulphate-dependent anaerobic oxidation of methane by marine microbes within a discrete sedimentary horizon termed the sulphate–methane transition zone. According to conventional isotope systematics, the biological consumption of methane leaves a residue of methane enriched in 13 C (refs 1–3). However, in many instances the methane within sulphate–methane transition zones is depleted in 13 C, consistent with the production of methane, and interpreted as evidence for the intertwined anaerobic oxidation and production of methane 4–6 . Here, we report results from experiments in which we incubated cultures of microbial methane consumers with methane and low levels of sulphate, and monitored the stable isotope composition of the methane and dissolved inorganic carbon pools over time. Residual methane became progressively enriched in 13 C at sulphate concentrations above 0.5 mM, and progressively depleted in 13 C below this threshold. We attribute the shift to 13 C depletion during the anaerobic oxidation of methane at low sulphate concentrations to the microbially mediated carbon isotope equilibration between methane and carbon dioxide. We suggest that this isotopic e ect could help to explain the 13 C-depletion of methane in subseafloor sulphate–methane transition zones.