Acetate Mineralization Research Articles

Sulfidic acetate mineralization at 45°C by an aquifer microbial community: key players and effects of heat changes on activity and community structure.

High-Temperature Aquifer Thermal Energy Storage (HT-ATES) is a sustainable approach for integrating thermal energy from various sources into complex energy systems. Temperatures ≥45°C, which are relevant in impact zones of HT-ATES systems, may dramatically influence the structure and activities of indigenous aquifer microbial communities. Here, we characterized an acetate-mineralizing, sulfate-reducing microbial community derived from an aquifer and adapted to 45°C. Acetate mineralization was strongly inhibited at temperatures ≤25°C and 60°C. Prolonged incubation at 12°C and 25°C resulted in acetate mineralization recovery after 40-80 days whereas acetate was not mineralized at 60°C within 100 days. Cultures pre-grown at 45°C and inhibited for 28 days by incubation at 12°C, 25°C, or 60°C recovered quickly after changing the temperature back to 45°C. Phylotypes affiliated to the order Spirochaetales and to endospore-forming sulfate reducers of the order Clostridiales were highly abundant in microcosms being active at 45°C highlighting their key role. In summary, prolonged incubation at 45°C resulted in active microbial communities mainly consisting of organisms adapted to temperatures between the typical temperature range of mesophiles and thermophiles and being resilient to temporary heat changes.

Gradual development of ammonia-induced syntrophic acetate-oxidizing activities under mesophilic and thermophilic conditions quantitatively tracked using multiple isotopic approaches

Insights into microbiota adaptation to increased ammonia stress, and identification of indicator microorganisms can help to optimize the operation of anaerobic digesters. To identify microbial indicators and investigate their metabolic contribution to acetoclastic methanogenesis (AM), syntrophic acetate oxidation (SAO) or hydrogenotrophic methanogenesis (HM), 40 anaerobic batch reactors fed with acetate of 110 mmol/L were set up at NH4+-N concentrations of 0.14 g/L, 5.00 g/L or 7.00 g/L, inoculated with thermophilic or mesophilic microbiota with or without pre-exposure to ammonia stress. Four stable carbon isotope probing approaches were applied in parallel, with [1,2-13C]-CH3COOH, [2-13C]-CH3COOH, [13C]NaHCO3 or non-labeled CH3COOH used individually. The last three approaches were used to quantify the methanogenic pathways by tracking labeled 13C or natural 13C signatures in the resulting CH4 and CO2, and consistently detected the dynamic transition of dominant pathways from AM to SAO-HM under ammonia stress. Results of quantitative PCR and fluorescence in-situ hybridization illustrated the procedure, acetotrophic methanogens being outcompeted by acetate-oxidizing syntrophs. The first and last isotope-labeling approaches were designed to probe the active acetate-mineralizing microbes with DNA-SIP. Known acetate-oxidizing bacteria like Syntrophaceticus and Tepidanaerobacter, as well as novel members of Pseudomonas, Bacillus and Symbiobacteraceae were detected, with Methanoculleus as the predominant H2/CO2-utilizing partner. Using NanoSIMS, some bacterial cells were observed to be fixing CO2 from [13C]NaHCO3. In this study, Methanosaeta was only active with ammonia < 200 mg-N/L; the syntrophs catalyzing SAO-HM started to compete with AM-conducting Methanosarcina at intermediate concentrations of ammonia, i.e. 200–500 mg-N/L, and outcompeted the acetotrophic methanogens with ammonia > 500 mg-N/L. Under ammonia stress, diverse known and novel microbial taxa were involved in acetate mineralization, comparable with those identified in previous studies.

Temperature management potentially affects carbon mineralization capacity and microbial community composition of a shallow aquifer.

High-temperature aquifer thermal energy storage (HT-ATES) is a promising technique to reduce the CO2 footprint of heat supply in the frame of transitioning to renewable energies. However, HT-ATES causes temperature fluctuations in groundwater ecosystems potentially affecting important microbial-mediated ecosystem services. Hence, assessing the impact of increasing temperatures on the structure and functioning of aquifer microbiomes is crucial to evaluate potential environmental risks associated with HT-ATES. In this study, we investigated the effects of temperature variations (12-80°C) on microbial communities and their capacity to mineralize acetate in aerobically incubated sediment sampled from a pristine aquifer. Compared to natural conditions (12°C), increased acetate mineralization rates were observed at 25°C, 37°C and 45°C, whereas mineralization was decelerated at 60°C and absent at 80°C. Sequencing of 16S rRNA genes revealed that the bacterial diversity in acetate-amended and non-acetate-amended sediments decreased with rising temperatures. Distinct communities dominated by bacterial groups affiliated with meso- and thermophilic bacteria established at 45°C and 60°C, respectively, while the number of archaeal phylotypes decreased. The changes in microbial diversity observed at 45°C and 60°C indicate a potential loss of ecosystem functioning, functional redundancy and resilience, while heat storage at 80°C bears the risk of ecological collapse.

Effect of Temperature on Acetate Mineralization Kinetics and Microbial Community Composition in a Hydrocarbon-Affected Microbial Community During a Shift From Oxic to Sulfidogenic Conditions.

Aquifer thermal energy storage (ATES) allows for the seasonal storage and extraction of heat in the subsurface thus reducing reliance on fossil fuels and supporting decarbonization of the heating and cooling sector. However, the impacts of higher temperatures toward biodiversity and ecosystem services in the subsurface environment remain unclear. Here, we conducted a laboratory microcosm study comprising a hydrocarbon-degrading microbial community from a sulfidic hydrocarbon-contaminated aquifer spiked with 13C-labeled acetate and incubated at temperatures between 12 and 80°C to evaluate (i) the extent and rates of acetate mineralization and (ii) the resultant temperature-induced shifts in the microbial community structure. We observed biphasic mineralization curves at 12, 25, 38, and 45°C, arising from immediate and fast aerobic mineralization due to an initial oxygen exposure, followed by slower mineralization at sulfidogenic conditions. At 60°C and several replicates at 45°C, acetate was only aerobically mineralized. At 80°C, no mineralization was observed within 178 days. Rates of acetate mineralization coupled to sulfate reduction at 25 and 38°C were six times faster than at 12°C. Distinct microbial communities developed in oxic and strictly anoxic phases of mineralization as well as at different temperatures. Members of the Alphaproteobacteria were dominant in the oxic mineralization phase at 12–38°C, succeeded by a more diverse community in the anoxic phase composed of Deltaproteobacteria, Clostridia, Spirochaetia, Gammaproteobacteria and Anaerolinea, with varying abundances dependent on the temperature. In the oxic phases at 45 and 60°C, phylotypes affiliated to spore-forming Bacilli developed. In conclusion, temperatures up to 38°C allowed aerobic and anaerobic acetate mineralization albeit at varying rates, while mineralization occurred mainly aerobically between 45 and 60°C; thermophilic sulfate reducers being active at temperatures > 45°C were not detected. Hence, temperature may affect dissolved organic carbon mineralization rates in ATES while the variability in the microbial community composition during the transition from micro-oxic to sulfidogenic conditions highlights the crucial role of electron acceptor availability when combining ATES with bioremediation.

Linking microbial Sphagnum degradation and acetate mineralization in acidic peat bogs: from global insights to a genome-centric case study.

Ombrotrophic bogs accumulate large stores of soil carbon that eventually decompose to carbon dioxide and methane. Carbon accumulates because Sphagnum mosses slow microbial carbon decomposition processes, leading to the production of labile intermediate compounds. Acetate is a major product of Sphagnum degradation, yet rates of hydrogenotrophic methanogenesis far exceed rates of aceticlastic methanogenesis, suggesting that alternative acetate mineralization processes exist. Two possible explanations are aerobic respiration and anaerobic respiration via humic acids as electron acceptors. While these processes have been widely observed, microbial community interactions linking Sphagnum degradation and acetate mineralization remain cryptic. In this work, we use ordination and network analysis of functional genes from 110 globally distributed peatland metagenomes to identify conserved metabolic pathways in Sphagnum bogs. We then use metagenome-assembled genomes (MAGs) from McLean Bog, a Sphagnum bog in New York State, as a local case study to reconstruct pathways of Sphagnum degradation and acetate mineralization. We describe metabolically flexible Acidobacteriota MAGs that contain all genes to completely degrade Sphagnum cell wall sugars under both aerobic and anaerobic conditions. Finally, we propose a hypothetical model of acetate oxidation driven by changes in peat redox potential that explain how bogs may circumvent aceticlastic methanogenesis through aerobic and humics-driven respiration.

Actinorhizal Alder Phytostabilization Alters Microbial Community Dynamics in Gold Mine Waste Rock from Northern Quebec: A Greenhouse Study.

Phytotechnologies are rapidly replacing conventional ex-situ remediation techniques as they have the added benefit of restoring aesthetic value, important in the reclamation of mine sites. Alders are pioneer species that can tolerate and proliferate in nutrient-poor, contaminated environments, largely due to symbiotic root associations with the N2-fixing bacteria, Frankia and ectomycorrhizal (ECM) fungi. In this study, we investigated the growth of two Frankia-inoculated (actinorhizal) alder species, A. crispa and A. glutinosa, in gold mine waste rock from northern Quebec. Alder species had similar survival rates and positively impacted soil quality and physico-chemical properties in similar ways, restoring soil pH to neutrality and reducing extractable metals up to two-fold, while not hyperaccumulating them into above-ground plant biomass. A. glutinosa outperformed A. crispa in terms of growth, as estimated by the seedling volume index (SVI), and root length. Pyrosequencing of the bacterial 16S rRNA gene for bacteria and the ribosomal internal transcribed spacer (ITS) region for fungi provided a comprehensive, direct characterization of microbial communities in gold mine waste rock and fine tailings. Plant- and treatment-specific shifts in soil microbial community compositions were observed in planted mine residues. Shannon diversity and the abundance of microbes involved in key ecosystem processes such as contaminant degradation (Sphingomonas, Sphingobium and Pseudomonas), metal sequestration (Brevundimonas and Caulobacter) and N2-fixation (Azotobacter, Mesorhizobium, Rhizobium and Pseudomonas) increased over time, i.e., as plants established in mine waste rock. Acetate mineralization and most probable number (MPN) assays showed that revegetation positively stimulated both bulk and rhizosphere communities, increasing microbial density (biomass increase of 2 orders of magnitude) and mineralization (five-fold). Genomic techniques proved useful in investigating tripartite (plant-bacteria-fungi) interactions during phytostabilization, contributing to our knowledge in this field of study.

Evaluation of the ecotoxicological impact of the organochlorine chlordecone on soil microbial community structure, abundance, and function.

The insecticide chlordecone applied for decades in banana plantations currently contaminates 20,000 ha of arable land in the French West Indies. Although the impact of various pesticides on soil microorganisms has been studied, chlordecone toxicity to the soil microbial community has never been assessed. We investigated in two different soils (sandy loam and silty loam) exposed to different concentrations of CLD (D0, control; D1 and D10, 1 and 10 times the agronomical dose) over different periods of time (3, 7, and 32 days): (i) the fate of chlordecone by measuring (14)C-chlordecone mass balance and (ii) the impact of chlordecone on microbial community structure, abundance, and function, using standardized methods (-A-RISA, taxon-specific quantitative PCR (qPCR), and (14)C-compounds mineralizing activity). Mineralization of (14)C-chlordecone was inferior below 1 % of initial (14)C-activity. Less than 2 % of (14)C-activity was retrieved from the water-soluble fraction, while most of it remained in the organic-solvent-extractable fraction (75 % of initial (14)C-activity). Only 23 % of the remaining (14)C-activity was measured in nonextractable fraction. The fate of chlordecone significantly differed between the two soils. The soluble and nonextractable fractions were significantly higher in sandy loam soil than in silty loam soil. All the measured microbiological parameters allowed discriminating statistically the two soils and showed a variation over time. The genetic structure of the bacterial community remained insensitive to chlordecone exposure in silty loam soil. In response to chlordecone exposure, the abundance of Gram-negative bacterial groups (β-, γ-Proteobacteria, Planctomycetes, and Bacteroidetes) was significantly modified only in sandy loam soil. The mineralization of (14)C-sodium acetate and (14)C-2,4-D was insensitive to chlordecone exposure in silty loam soil. However, mineralization of (14)C-sodium acetate was significantly reduced in soil microcosms of sandy loam soil exposed to chlordecone as compared to the control (D0). These data show that chlordecone exposure induced changes in microbial community taxonomic composition and function in one of the two soils, suggesting microbial toxicity of this organochlorine.

Carbon source preference in chemosynthetic hot spring communities.

Rates of dissolved inorganic carbon (DIC), formate, and acetate mineralization and/or assimilation were determined in 13 high-temperature (>73 °C) hot springs in Yellowstone National Park (YNP), Wyoming, in order to evaluate the relative importance of these substrates in supporting microbial metabolism. While 9 of the hot spring communities exhibited rates of DIC assimilation that were greater than those of formate and acetate assimilation, 2 exhibited rates of formate and/or acetate assimilation that exceeded those of DIC assimilation. Overall rates of DIC, formate, and acetate mineralization and assimilation were positively correlated with spring pH but showed little correlation with temperature. Communities sampled from hot springs with similar geochemistries generally exhibited similar rates of substrate transformation, as well as similar community compositions, as revealed by 16S rRNA gene-tagged sequencing. Amendment of microcosms with small (micromolar) amounts of formate suppressed DIC assimilation in short-term (<45-min) incubations, despite the presence of native DIC concentrations that exceeded those of added formate by 2 to 3 orders of magnitude. The concentration of added formate required to suppress DIC assimilation was similar to the affinity constant (K(m)) for formate transformation, as determined by community kinetic assays. These results suggest that dominant chemoautotrophs in high-temperature communities are facultatively autotrophic or mixotrophic, are adapted to fluctuating nutrient availabilities, and are capable of taking advantage of energy-rich organic substrates when they become available.

Microbiological Reduction of Sb(V) in Anoxic Freshwater Sediments

Microbiological reduction of millimolar concentrations of Sb(V) to Sb(III) was observed in anoxic sediments from two freshwater settings: (1) a Sb- and As-contaminated mine site (Stibnite Mine) in central Idaho and 2) an uncontaminated suburban lake (Searsville Lake) in the San Francisco Bay Area. Rates of Sb(V) reduction in anoxic sediment microcosms and enrichment cultures were enhanced by amendment with lactate or acetate as electron donors but not by H2, and no reduction occurred in sterilized controls. Addition of 2-(14)C-acetate to Stibnite Mine microcosms resulted in the production of (14)CO2 coupled to Sb(V) reduction, suggesting that this process proceeds by a dissimilatory respiratory pathway in those sediments. Antimony(V) reduction in Searsville Lake sediments was not coupled to acetate mineralization and may be associated with Sb-resistance. The microcosms and enrichment cultures also reduced sulfate, and the precipitation of insoluble Sb(III)-sulfide complexes was a major sink for reduced Sb. The reduction of Sb(V) by Stibnite Mine sediments was inhibited by As(V), suggesting that As(V) is a preferred electron acceptor for the indigenous community. These findings indicate a novel pathway for anaerobic microbiological respiration and suggest that communities capable of reducing high concentrations of Sb(V) commonly occur naturally in the environment.

Distribution of CO2 fixation and acetate mineralization pathways in microorganisms from extremophilic anaerobic biotopes

Extremophilic anaerobes are widespread in saline, acid, alkaline, and high or low temperature environments. Carbon is essential to living organisms and its fixation, degradation, or mineralization is driven by, up to now, six metabolic pathways. Organisms using these metabolisms are known as autotrophs, acetotrophs or carbon mineralizers, respectively. In anoxic and extreme environments, besides the well-studied Calvin-Benson-Bassham cycle, there are other five carbon fixation pathways responsible of autotrophy. Moreover, regarding carbon mineralization, two pathways perform this key process for carbon cycling. We might imagine that all the pathways can be found evenly distributed in microbial biotopes; however, in extreme environments, this does not occur. This manuscript reviews the most commonly reported anaerobic organisms that fix carbon and mineralize acetate in extreme anoxic habitats. Additionally, an inventory of autotrophic extremophiles by biotope is presented.

Activity and Distribution of Thermophilic Prokaryotes in Hydrothermal Fluid, Sulfidic Structures, and Sheaths of Alvinellids (East Pacific Rise, 13°N)

Processes of inorganic carbon assimilation, methanogenesis, sulfate reduction, and acetate oxidation to CO(2) occurring in samples from the East Pacific Rise at 13°N were traced, using radioisotopically labeled substrates, at temperatures ranging from 65 to 100°C. Molecular hydrogen stimulated lithotrophic methanogenesis and sulfate reduction but inhibited inorganic carbon assimilation. Active mineralization of acetate was observed in an organic-rich Alvinella-associated system at 80°C. Members of the Thermococcales were the most numerous hyperthermophilic archaea in these samples, their density achieving 10(8) cells per cm(3), while the numbers of cultured hydrogen-utilizing thermophilic lithotrophs were several orders of magnitude lower.

The application of a simplified method to map the aerobic acetate mineralization rates at the groundwater table of the Netherlands

A simplified method is used to assess the microbial activity of subsoils and soils across a broad geographic scale. Acetate was selected because it is a major intermediate in catabolic biochemical pathways. In order to get minimal disturbance, only a small amount of tritium labelled acetate and water is added to the subsoil material. After an incubation time, the subsoil material is separated from the water by centrifugation and the formed tritium labelled water is separated from the remaining acetate by evaporation. The data of 128 locations in the Netherlands were plotted in a soil map and were also compared with the depth, dry weight, electric conductivity, pH and nitrate concentration. The peat areas consisted of limed meadows with a high groundwater level whereas the sand areas often showed deeper groundwater levels and a lower pH. The subsoils at the groundwater table of the peat areas, which are in contact with soil air, showed a higher mineralization rate compared with the surface soils in our study. In contrast, the mineralization rate of the subsoil at the groundwater table of sandy soils showed on average a factor 30 lower rate. Nevertheless, the self purification capacity of the subsoil can be vital under weather conditions where the surface soil becomes less active.

Sulfide and ammonium oxidation, acetate mineralization by denitrification in a multipurpose UASB reactor

The physiological and kinetic behavior of a denitrifying granular sludge exposed to different sulfide loading rates (55-295 mg/L d) were evaluated in a UASB reactor fed with acetate, ammonium and nitrate. At any sulfide loading rates, the consumption efficiencies of sulfide, acetate and ammonium were above 95%, while nitrate consumption efficiencies were around 62-72%. At the highest sulfide loading rate the ammonium was used as electron donor for N(2) production. The increase of sulfide loading rate also affected the fate of sulfide oxidation, since elemental sulfur was the main end product instead of sulfate. However, the lithotrophic denitrifying kinetic was not affected. FISH oligonucleotide probes for Thiobacillus denitrificans, Thiomiscropira denitrificans, genus Paracoccus and Pseudomonas spp. were used to follow the microbial ecology. The results of this work have shown that four pollutants could simultaneously be removed, namely, sulfide, ammonium, acetate and nitrate under well defined denitrifying conditions.

Organic compounds that reach subsoil may threaten groundwater quality; effect of benzotriazole on degradation kinetics and microbial community composition

Toxic compounds in soils threaten groundwater quality in two ways: as potential contaminants themselves, and by retarding the microbial degradation of other organic compounds, thus enhancing their deep penetration. Benzotriazole (BTA) is a chemical with versatile industrial applications, used in large quantities worldwide, and represents a potential threat to the environment due to its apparent toxicity and recalcitrance. When used as an additive in aircraft deicing/antiicing fluid on airports, substantial spills of these mixtures and jet fuel will inevitably reach the soil. We have investigated the subsoil (1–2 m depth) microbial degradation and growth on four relevant organic substrates found in airport run-off (acetate, formate, glycol and toluene) in the presence of concentrations of BTA which can be found in airport run-off. Monitoring CO 2 evolution showed growth-dependent degradation rates for all substrates (sigmoid CO 2 accumulation curves), which were significantly affected by BTA. The mineralization of acetate was only moderately retarded and only by the highest BTA concentration used (400 mg l −1 in soil solution); formate and glycol mineralization was substantially retarded at 200 mg l −1, and toluene mineralization already at 10 mg l −1 BTA. Mass balances (fraction of added C recovered as CO 2) suggested that the microbial growth yield (g biomass-C formed per g substrate C) was severely reduced with increasing concentrations of BTA. The analysis of phospholipid fatty acids (PLFA) demonstrated that Gram-negative bacteria were dominating among the organisms growing on all four substrates. The total amount of PLFA increased with approximately 1000 pmol PLFA g −1 soil in response to a dose of 0.93 μmol glycol-C g −1 soil, but this increase was gradually reduced with increasing BTA concentrations. This was in agreement with C mass balances based on CO 2 measurements, verifying that BTA severely reduced the growth yields. The response of individual PLFA's to BTA and substrates demonstrated that non-growing organisms were largely unaffected (i.e. the PLFA's of which the absolute amounts did not increase in response to substrates were not affected by BTA), whereas those which were growing on the added substrates were uniformly reduced by BTA (all the PLFA's which increased in response to the substrates were negatively affected by BTA). The results suggest that BTA functions as an uncoupler, i.e. a substance that reduces the yield of ATP per mole of substrate used, or that the defence mechanisms represent a large energy burden to all microbial cells.

Dynamics and influencing factors of heterotrophic bacterial utilization of acetate in constructed wetlands treating woodwaste leachate

14C-acetate was used as a tracer of substrates for heterotrophic bacteria to investigate the dynamics and influencing factors of bacterial activity in surface flow wetlands treating woodwaste leachate. Epiphytic assimilation and mineralization of acetate increased considerably over three weeks of microbial colonization. No significant longitudinal variation in acetate uptake was found. The temporal variation of bacterial activity was neither correlated to water temperature nor acetic acid concentration. Mineralization percentage varied significantly with acetate uptake rate. Acetate uptake rates of bacterioplankton (408±258 μg L −1 h −1), epiphyton (67.7±41.7 mg m −2 h −1) and sedimentary bacteria (26.7±25.6 μg g −1 h −1) were influenced by the concentrations of organic substrates and inorganic nutrients. Sedimentary bacteria contributed to the majority (55–73%) of the total heterotrophic acetate uptake, while epiphytic bacteria played only a minor role (1–3%). The acetate mineralization percentage of sediment (16%) was much lower than that of water (55%) and epiphyton (64%). Addition of ammonium nitrate fertilizer increased acetate uptake rates significantly, especially for sediment. No remarkable changes in mineralization percentage and the relative importance of water, sediment and epiphyton were found due to fertilization.

The relation between extrapolated risk, expressed as potentially affected fraction, and community effects, expressed as pollution‐induced community tolerance

The results of toxicity tests can be used to calculate the potentially affected fraction (PAF) of species in an ecosystem at a given pollutant concentration using statistical extrapolation methods. The PAF curve indicates the fraction of species from the original community that may become inhibited at each elevated pollutant concentration and is a measure of the ecotoxicological risk. Pollution-induced community tolerance (PICT) is a true community response that is measured under controlled conditions in the laboratory, using organisms from contaminated field sites. Microorganisms from experimental field plots with added Zn were exposed to various concentrations of Zn in the laboratory and the mineralization of 14C acetate was monitored. Microorganisms from plots with Zn concentrations above 124 mg/kg showed a significant increase in the effect concentration 10% (EC10) and, therefore, had a significant PICT. The pore-water concentrations of Zn in these field soils were in the same magnitude as the EC10 of the microorganisms from these soils. The PAF curve was calculated from previously reported toxicity tests with five different microbial species using the average and the standard deviation of the logarithmically transformed EC10 values. The average sensitivity of this PAF curve was similar to the EC50 of the acetate mineralization curve from the field plot without added Zn2+, but the PAF curve was less steep. Our experiments indicated that 27 to 84% of the original microbial species were inhibited at Zn concentrations from 334 to 1,858 mg/kg soil, respectively. Our results suggest that the PICT method can now also be used to quantify the fraction of the original species composition that is inhibited at a specific pollutant concentration.

THE RELATION BETWEEN EXTRAPOLATED RISK, EXPRESSED AS POTENTIALLY AFFECTED FRACTION, AND COMMUNITY EFFECTS, EXPRESSED AS POLLUTION-INDUCED COMMUNITY TOLERANCE

The results of toxicity tests can be used to calculate the potentially affected fraction (PAF) of species in an ecosystem at a given pollutant concentration using statistical extrapolation methods. The PAF curve indicates the fraction of species from the original community that may become inhibited at each elevated pollutant concentration and is a measure of the ecotoxicological risk. Pollution-induced community tolerance (PICT) is a true community response that is measured under controlled conditions in the laboratory, using organisms from contaminated field sites. Microorganisms from experimental field plots with added Zn were exposed to various concentrations of Zn in the laboratory and the mineralization of 14C acetate was monitored. Microorganisms from plots with Zn concentrations above 124 mg/kg showed a significant increase in the effect concentration 10% (EC10) and, therefore, had a significant PICT. The pore-water concentrations of Zn in these field soils were in the same magnitude as the EC10 of the microorganisms from these soils. The PAF curve was calculated from previously reported toxicity tests with five different microbial species using the average and the standard deviation of the logarithmically transformed EC10 values. The average sensitivity of this PAF curve was similar to the EC50 of the acetate mineralization curve from the field plot without added Zn2+, but the PAF curve was less steep. Our experiments indicated that 27 to 84% of the original microbial species were inhibited at Zn concentrations from 334 to 1,858 mg/kg soil, respectively. Our results suggest that the PICT method can now also be used to quantify the fraction of the original species composition that is inhibited at a specific pollutant concentration.

A comparison between toxicity tests using single species and a microbial process

In this study the sensitivity of the acetate mineralization process performed by five strains of micro-organisms in soil for the toxicants Zn 2+ or PCP was calculated from the sensitivity of the contributing species. The species used were a fungus ( Aspergillus niger CBS 121.49), an actinomycete ( Streptomyces lividans 66), two Gram-negative Pseudomonas putida strains (MT-2 and DSM 50026) and a Gram-positive strain Rhodococcus erythropolis A177. For zinc the EC10 of the process performed by the five strains together was 77 mg/kg whereas for pentachlorophenol it was 2 mg/kg. The EC10 of the process was compared with the EC50 of the most sensitive species contributing to the process. P. putida MT2 was the most zinc sensitive strain (EC50 = 22 mg Zn/kg) and A. niger was the most sensitive strain for pentachlorophenol (EC50 = 1.4 mg/kg). This shows that a 10% inhibition of a process can be accompanied by a more than 50% inhibition of the most sensitive species.

Acetate Fate in Organic and Conventionally Managed Soils

AbstractOrganic management practices and mixed‐crop rotations have beneficial impacts on biologically active soil organic matter (SOM); however, the specific impacts of these practices on SOM composition and turnover characteristics are not known. The effect of organic and conventional management on soil C cycling was assessed using soils collected from Rodale's Farming Systems Trial experiment, which included organic‐manure amended, organic‐legume cover cropped, and conventionally managed‐inorganically fertilized farming systems. Soil respiration rates, atom % 13C, and cross polarization 13C nuclear magnetic resonance (CP‐MAS 13C NMR) were used to trace the fate of 13C‐labeled acetate added to soils incubated for 2 and 13 wk. Microbial activity and C turnover rates were initially greater in the manure‐amended soil than in the other two soils. Acetate mineralization lagged in the cover‐cropped soil during the first 2 wk. Low atom % 13C in the conventional soil showed that, even though its soil respiration rates, and therefore overall microbial activity, equaled rates in the cover‐cropped soil, microorganisms present in this soil metabolized significantly more of the applied acetate. By the end of the experiment, C cycling characteristics of the cover‐cropped soil were similar to the manure‐amended soil; NMR spectra indicated 13C was incorporated into new SOM in both of these soils. Soil respiration and 13C assimilation rates in the conventional soil remained relatively low. Microorganisms in the organic soils mineralized and respired more native C and assimilated more 13C than the conventional soil. Conversely, the conventional soil mineralized more and assimilated less 13C than the organic soils. Differences observed in this laboratory experiment may or may not reflect on C cycling occurring in the field.

Rapid Toluene Mineralization by Aquifer Microorganisms at Adak, Alaska: Implications for Intrinsic Bioremediation in Cold Environments

Sediments from a relatively cold (5 °C), petroleum hydrocarbon-contaminated aquifer in Adak, AK, mineralized [ 14 C]toluene at an aerobic rate (16.3% day -1 at 5 °C) comparable to that (5.1% day -1 at 20 °C) of sediments from a more temperate aquifer at Hanahan, SC. In addition, rates of overall microbial metabolism in sediments from the two aquifers, as estimated by [1- 14 C]acetate mineralization, were similar (∼10.6% h -1 ) at their respective in situ temperatures. These results are not consistent with the common assumption that biodegradation rates in cold groundwater systems are depressed relative to more temperate systems. Furthermore, these results suggest that intrinsic bioremediation of petroleum hydrocarbon contaminants in cold groundwater systems may be technically feasible, in some cases.