Indigenous Sediment Microbial Activity in Response to Nutrient Enrichment and Plant Growth Following a Controlled Oil Spill on a Freshwater Wetland

The population density and activity of a microbial community associated with the sediment and rhizosphere of an intertidal freshwater wetland dominated by Scirpus pungens was monitored before and following the application of weathered Mesa light crude oil and fertilizers. The influence of nutrient enrichment (fertilizers) and plant growth on oil degradation rates was determined from the resulting data. The study plots (four blocks of replicates) were subjected to five treatments: oil only (natural attenuation); oil plus ammonium nitrate and phosphate, with regular cropping of the plants; oil plus ammonium nitrate and phosphate; oil plus sodium nitrate and phosphate; no oil, ammonium nitrate and phosphate. The plots were regularly monitored in the field for gas production (carbon dioxide and nitrous oxide), and samples were collected for laboratory analysis of denitrification activity, aliphatic and aromatic hydrocarbon degradation activity, and total heteroptrophic bacteria. The viable bacterial population density increased during the first 4 weeks in oiled and unoiled experimental plots that were fertilized. In contrast, population densities in untreated areas remained relatively unchanged throughout the monitoring period. The microbial population demonstrated a rapid and sustained increase in naphthalene mineralization activity in plots that were both fertilized and oiled. Hexadecane mineralization activity increased in response to fertilizer application, with ammonium nitrate causing a larger increase than sodium nitrate. A very significant difference observed in the mineralization of hexadecane was that the surface sediments were much more active than the subsurface sediments. This difference became even more pronounced in the second year of monitoring, even though the treatment regime had been discontinued. This compartmentalization of mineralization activity was not observed for naphthalene. Following fertilizer application, field and laboratory evaluation of nitrogen metabolism in the sediments indicated significant denitrification activity that was not adversely affected by oiling. The results demonstrated that the application of fertilizers stimulated the activities of indigenous hydrocarbon-degrading and denitrifying bacteria, and the presence of oil either enhanced or had no detrimental effect on these activities. As a remediation strategy, the application of fertilizers to a wetland shoreline following an oil spill would promote the growth of indigenous plants and their associated microbial flora, resulting in increased metabolic activity and the potential for increased oil degradation activity.     

Bioavailability to Fish of Sediment PAH as an Indicator of the Success of In Situ Remediation Treatments at an Experimental Oil Spill

A weathered medium crude oil was applied to experimental plots of Scirpus pungens (Three-square Bulrush) in a freshwater wetland to determine the efficacy of strategies for shoreline oil spill bioremediation based on nutrient enrichment (bioremediation) and plant growth (phytoremediation). Plots were unoiled, oiled with no added nutrients, or oiled with repeated applications of phosphate and nitrate fertilizers. Following initial treatments, the experimental plots were raked to simulate the activity of wave action on oil penetration, and plants in one fertilized plot were cut repeatedly. The sediments were sampled at regular intervals for 15 months after oiling, and the loss of oil was assessed by 4-day laboratory tests of polynuclear aromatic hydrocarbon (PAH) bioaccumulation by trout, as demonstrated by increases in activity of liver cytochrome P450 (CYP1A) enzymes. Oil alone, oil mixed with sediments in the lab, and oiled sediments from treated plots all induced CYP1A activity relative to untreated controls, indicating the presence and bioavailability of PAH. Induction did not vary with nutrient treatments, but declined by 80% within 15 months of oiling, and chemical analyses indicated equivalent losses of hydrocarbons in sediment. These results demonstrate that bioavailable PAHs persisted in measurable quantities for at least 1.25 years following oiling, and that stimulation of plant growth did not affect the rate of oil disappearance. The controlling factors were likely weathering and sediment movement.     

Microbial Dynamics during Bioremediation of a Crude Oil-Contaminated Coastal Wetland

In 1996, a controlled crude oil application was conducted at a Texas intertidal, coastal wetland to determine the effectiveness of two biostimulation treatments in these sensitive areas. An inorganic nutrient treatment and inorganic nutrient plus a potential electron acceptor (nitrate) treatment were examined. As part of this research, polycyclic aromatic hydrocarbon (PAH)-degrading, aliphatic-degrading, and total heterotrophic microbial numbers were monitored. Using a randomized, complete block design consisting of 21 plots, microbial data from biostimulation treatment plots were statistically compared to oiled control plots to assess treatment differences. Sediment samples from all plots receiving oil showed exponential increases in the numbers of aliphatic (n-alkane) and PAH-degrading microorganisms. This increase was observed at both 0 to 5 cm and 5 to 10 cm sample depths. Statistical analysis, however, revealed no significant differences in the numbers of aliphatic-degrading or PAH-degrading microorganisms between treatment plots and oiled control plots or between treatments on any sample day. The numbers of PAH- and aliphatic-degrading microorganisms returned to near pre-application levels by the end of the monitoring period. Ratios of hydrocarbon-degrading microbes to total heterotrophs also increased as a result of the oil application and returned to pre-application levels by the end of the monitoring period. Overall, the populations of hydrocarbon-degrading microorganisms illustrated a well-documented response to crude oil. However, the addition of the biostimulation treatments did not significantly increase the numbers of aliphatic-degrading, PAH-degrading, or total heterotrophic microorganisms over populations on control plots.     

Microbial Population Analysis as a Measure of Ecosystem Restoration

During a controlled oil spill study in a freshwater wetland, four methods were used to track changes in microbial populations in response to in situ remediation treatments, including nutrient amendments and the removal of surface vegetation. Most probable number (MPN) estimates of alkane and aromatic hydrocarbon degraders showed divergence of the alkane and aromatic degrading populations during the first summer of the experiment. Alkane degraders increased in all plots by 1.5 orders of magnitude and aromatic degraders increased in oiled plots by 3.5 orders of magnitude. Phospholipid fatty acid (PLFA) analysis of biomass and community composition showed no essential differences among treatments. Denaturing gradient gel electrophoresis (DGGE) analysis of the sediment microbial community showed some differences in specific populations of organisms with respect to oiled and unoiled plots. Some organisms were only found in the oiled plots. Sediment toxicity measured against single celled algae showed that the oiled sediments were toxic into the second year of the study, but that nutrient addition relieved the toxicity more rapidly than natural attenuation of the oil.     

Microbial populations and hydrocarbon biodegradation potentials in fertilized shoreline sediments affected by the T/V Exxon Valdez oil spill.

The effort of clean up the T/V Exxon Valdez oil spill in Prince William Sound, Alaska, included the use of fertilizers to accelerate natural microbial degradation of stranded oil. A program to monitor various environmental parameters associated with this technique took place during the summer of 1990. Microbiological assays for numbers of heterotrophic and oil-degrading microbes and their hydrocarbon mineralization potentials were performed in support of this program. Fertilizer addition resulted in higher hexadecane and phenanthrene mineralization potentials on treated plots than on untreated reference plots. Microbial numbers in treated and reference surface sediments were not significantly different immediately after the first nutrient application in May 1990. However, subsurface sediments from treated plots had higher numbers of hydrocarbon degraders than did reference sediments shortly after treatment. The second application of fertilizer, later in summer, resulted in surface and subsurface increases in numbers of hydrocarbon degraders with respect to reference sediments at two of the three study sites. Elevated mineralization potentials, coupled with increased numbers of hydrocarbon degraders, indicated that natural hydrocarbon biodegradation was enhanced. However, these microbiological measurements alone are not sufficient to determine in situ rates of crude oil biodegradation.     

Microbial populations and hydrocarbon biodegradation potentials in fertilized shoreline sediments affected by the T/V Exxon Valdez oil spill.

The effort of clean up the T/V Exxon Valdez oil spill in Prince William Sound, Alaska, included the use of fertilizers to accelerate natural microbial degradation of stranded oil. A program to monitor various environmental parameters associated with this technique took place during the summer of 1990. Microbiological assays for numbers of heterotrophic and oil-degrading microbes and their hydrocarbon mineralization potentials were performed in support of this program. Fertilizer addition resulted in higher hexadecane and phenanthrene mineralization potentials on treated plots than on untreated reference plots. Microbial numbers in treated and reference surface sediments were not significantly different immediately after the first nutrient application in May 1990. However, subsurface sediments from treated plots had higher numbers of hydrocarbon degraders than did reference sediments shortly after treatment. The second application of fertilizer, later in summer, resulted in surface and subsurface increases in numbers of hydrocarbon degraders with respect to reference sediments at two of the three study sites. Elevated mineralization potentials, coupled with increased numbers of hydrocarbon degraders, indicated that natural hydrocarbon biodegradation was enhanced. However, these microbiological measurements alone are not sufficient to determine in situ rates of crude oil biodegradation.     

Microbial degradation of acenaphthene and naphthalene under denitrification conditions in soil-water systems

This study examined the microbial degradation of acenaphthene and naphthalene under denitrification conditions at soil-to-water ratios of 1:25 and 1:50 with soil containing approximately 10(5) denitrifying organisms per g of soil. Under nitrate-excess conditions, both acenaphthene and naphthalene were degraded from initial aqueous-phase concentrations of about 1 and several mg/liter respectively, to nondetectable levels (less than 0.01 mg/liter) in less than 9 weeks. Acclimation periods of 12 to 36 days were observed prior to the onset of microbial degradation in tests with soil not previously exposed to polycyclic aromatic hydrocarbon (PAH) compounds, whereas acclimation periods were absent in tests with soil reserved from prior PAH degradation tests. It was judged that the apparent acclimation period resulted from the time required for a small population of organisms capable of PAH degradation to attain sufficient densities to exhibit detectable PAH reduction, rather than being a result of enzyme induction, mutation, or use of preferential substrate. About 0.9% of the naturally occurring soil organic carbon could be mineralized under denitrification conditions, and this accounted for the greater proportion of the nitrate depletion. Mineralization of the labile fraction of the soil organic carbon via microbial denitrification occurred without an observed acclimation period and was rapid compared with PAH degradation. Under nitrate-limiting conditions the PAH compounds were stable owing to the depletion of nitrate via the more rapid process of soil organic carbon mineralization. Soil sorption tests showed at the initiation of a test that the total mass of PAH compound was divided in comparable proportions between solute in the aqueous phase and solute sorbed on the solid phase. The microbial degradation of the PAH compound depends on the interrelationships between (i) the desorption kinetics and the reversibility of desorption of sorbed compound from the soil, (ii) the concentration of PAH-degrading microorganisms, and (iii) the competing reaction for nitrate utilization via mineralization of the labile fraction of naturally occurring soil organic carbon.     

Microbial degradation of acenaphthene and naphthalene under denitrification conditions in soil-water systems.

This study examined the microbial degradation of acenaphthene and naphthalene under denitrification conditions at soil-to-water ratios of 1:25 and 1:50 with soil containing approximately 10(5) denitrifying organisms per g of soil. Under nitrate-excess conditions, both acenaphthene and naphthalene were degraded from initial aqueous-phase concentrations of about 1 and several mg/liter respectively, to nondetectable levels (less than 0.01 mg/liter) in less than 9 weeks. Acclimation periods of 12 to 36 days were observed prior to the onset of microbial degradation in tests with soil not previously exposed to polycyclic aromatic hydrocarbon (PAH) compounds, whereas acclimation periods were absent in tests with soil reserved from prior PAH degradation tests. It was judged that the apparent acclimation period resulted from the time required for a small population of organisms capable of PAH degradation to attain sufficient densities to exhibit detectable PAH reduction, rather than being a result of enzyme induction, mutation, or use of preferential substrate. About 0.9% of the naturally occurring soil organic carbon could be mineralized under denitrification conditions, and this accounted for the greater proportion of the nitrate depletion. Mineralization of the labile fraction of the soil organic carbon via microbial denitrification occurred without an observed acclimation period and was rapid compared with PAH degradation. Under nitrate-limiting conditions the PAH compounds were stable owing to the depletion of nitrate via the more rapid process of soil organic carbon mineralization. Soil sorption tests showed at the initiation of a test that the total mass of PAH compound was divided in comparable proportions between solute in the aqueous phase and solute sorbed on the solid phase. The microbial degradation of the PAH compound depends on the interrelationships between (i) the desorption kinetics and the reversibility of desorption of sorbed compound from the soil, (ii) the concentration of PAH-degrading microorganisms, and (iii) the competing reaction for nitrate utilization via mineralization of the labile fraction of naturally occurring soil organic carbon.     

Structure and activity of the denitrifying community in a maize-cropped field fertilized with composted pig manure or ammonium nitrate

One alternative to mineral fertilization is to use organic fertilizers. Our aim was to compare the impacts of 7-year applications of composted pig manure and ammonium nitrate on the structure and activity of the denitrifying community. Mineralization and organization of N, denitrification rates and N2O/N2 ratio were also investigated. Fourteen months after the last application, the potential denitrifying activity (+319%), N mineralization (+110%) and organization (+112%) were higher under pig compost than under ammonium nitrate fertilization. On the other hand, the N2O/(N2O+N2) ratio was lower (P<0.05, n=5) under organic fertilization. These effects of organic fertilization were in accordance with its higher total carbon content and microbial biomass. Fingerprints and clone library analyses showed that the structure of the denitrifying community was affected by the fertilization regime. Our results reveal that organic or mineral fertilizer applications could affect both structure and activity of the denitrifying community, with a possible influence on in situ N2O fluxes. These effects of the fertilization regime persisted for at least 14 months after the last application.     

Microbial Population Changes during Bioremediation of an Experimental Oil Spill

Three crude oil bioremediation techniques were applied in a randomized block field experiment simulating a coastal oil spill. Four treatments (no oil control, oil alone, oil plus nutrients, and oil plus nutrients plus an indigenous inoculum) were applied. In situ microbial community structures were monitored by phospholipid fatty acid (PLFA) analysis and 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) to (i) identify the bacterial community members responsible for the decontamination of the site and (ii) define an end point for the removal of the hydrocarbon substrate. The results of PLFA analysis demonstrated a community shift in all plots from primarily eukaryotic biomass to gram-negative bacterial biomass with time. PLFA profiles from the oiled plots suggested increased gram-negative biomass and adaptation to metabolic stress compared to unoiled controls. DGGE analysis of untreated control plots revealed a simple, dynamic dominant population structure throughout the experiment. This banding pattern disappeared in all oiled plots, indicating that the structure and diversity of the dominant bacterial community changed substantially. No consistent differences were detected between nutrient-amended and indigenous inoculum-treated plots, but both differed from the oil-only plots. Prominent bands were excised for sequence analysis and indicated that oil treatment encouraged the growth of gram-negative microorganisms within the α-proteobacteria and Flexibacter-Cytophaga-Bacteroides phylum. α-Proteobacteria were never detected in unoiled controls. PLFA analysis indicated that by week 14 the microbial community structures of the oiled plots were becoming similar to those of the unoiled controls from the same time point, but DGGE analysis suggested that major differences in the bacterial communities remained.     

Microbial population changes during bioremediation of an experimental oil spill.

Three crude oil bioremediation techniques were applied in a randomized block field experiment simulating a coastal oil spill. Four treatments (no oil control, oil alone, oil plus nutrients, and oil plus nutrients plus an indigenous inoculum) were applied. In situ microbial community structures were monitored by phospholipid fatty acid (PLFA) analysis and 16S rDNA PCR-denaturing gradient gel electrophoresis (DGGE) to (i) identify the bacterial community members responsible for the decontamination of the site and (ii) define an end point for the removal of the hydrocarbon substrate. The results of PLFA analysis demonstrated a community shift in all plots from primarily eukaryotic biomass to gram-negative bacterial biomass with time. PLFA profiles from the oiled plots suggested increased gram-negative biomass and adaptation to metabolic stress compared to unoiled controls. DGGE analysis of untreated control plots revealed a simple, dynamic dominant population structure throughout the experiment. This banding pattern disappeared in all oiled plots, indicating that the structure and diversity of the dominant bacterial community changed substantially. No consistent differences were detected between nutrient-amended and indigenous inoculum-treated plots, but both differed from the oil-only plots. Prominent bands were excised for sequence analysis and indicated that oil treatment encouraged the growth of gram-negative microorganisms within the alpha-proteobacteria and Flexibacter-Cytophaga-Bacteroides phylum. alpha-Proteobacteria were never detected in unoiled controls. PLFA analysis indicated that by week 14 the microbial community structures of the oiled plots were becoming similar to those of the unoiled controls from the same time point, but DGGE analysis suggested that major differences in the bacterial communities remained.     

Nutrient retention and release in a floodplain's aquifer and in the hyporheic zone of a lowland river

Fertilizer applications and other non-point sources result in an increasing diffuse N and P pollution of receiving waters degrading water quality by eutrophication with several adverse impacts. Floodplains are regarded as reactive interfaces between uplands and receiving waters. In the present study groundwater quality on its subsurface flow from an upland area through a lowland floodplain towards the receiving water body of the Spree River was monitored biweekly over 2 years with two transects of 18 groundwater observation wells. Within the floodplain reaction rates of the nutrients are unevenly distributed. On a scale smaller than the floodplain, the hyporheic zone is regarded as reactive interface with unproportional high reaction rates. Therefore, phosphate and dissolved iron were measured with high spatial resolution in the pore water of the riverbed and the oxbow bed to investigate turnover processes and their small-scale spatial variability at the immediate surface–subsurface interface. The biogeochemical composition of subsurface water is characterized by little temporal variability while spatial heterogeneity is high on the hectametre scale of the study site as well as on the centimetre scale of the bed sediments. Nitrate is eliminated very efficiently by denitrification in the anoxic aquifer of the floodplain while ammonium and phosphate concentrations increase under anoxic conditions. Phosphate and ammonium originate from the mineralization of organic matter and phosphate is additionally released by reductive dissolution of iron-bound phosphorus and weathering of bedrock. Sorption–desorption processes equalize temporal fluctuations of phosphate concentrations. Phosphate uptake by plants is assumed as an important process at only one of the groundwater observation wells. Redox conditions required for a phosphate sink are opposite to those involved in nitrate removal by denitrification. Thus, redox patchiness of floodplain aquifers favours nitrate and phosphate removal, i.e. a temporal and spatial sequence of anoxic and oxic conditions eliminates nitrogen and causes phosphate storage. On the groundwater's path from the upland to the river further phosphate is released in the bed sediments. It originates from previously settled particulate compounds containing phosphorus. While the release of iron-bound phosphorus clearly predominates in the riverbed sediments the mineralization of organic matter is an important additional phosphorus release process in the oxbow bed sediments.     

The influence of an invasive plant on denitrification in an urban wetland

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Denitrification in Marl and Peat Sediments in the Florida Everglades

The potential for denitrification in marl and peat sediments in the Shark River Slough in the Everglades National Park was determined by the acetylene blockage assay. The influence of nitrate concentration on denitrification rate and N₂O yield from added nitrate was examined. The effects of added glucose and phosphate and of temperature on the denitrification potential were determined. The sediments readily denitrified added nitrate. N₂O was released from the sediments both with and without added acetylene. The marl sediments had higher rates than the peat on every date sampled. Denitrification was nitrate limited; however, the yields of N₂O amounted to only 10 to 34% of the added nitrate when 100 μM nitrate was added. On the basis of measured increases in ammonium concentration, it appears that the balance of added nitrate may be converted to ammonium in the marl sediment. The sediment temperature at the time of sampling greatly influenced the denitrification potential (15-fold rate change) at the marl site, indicating that either the number or the specific activity of the denitrifiers changed in response to temperature fluctuations (9 to 25°C) in the sediment. It is apparent from this study that denitrification in Everglades sediments is not an effective means of removing excess nitrogen which may be introduced as nitrate into the ecosystem with supply water from the South Florida watershed and that sporadic addition of nitrate-rich water may lead to nitrous oxide release from these wetlands.     

Soil amendments promote denitrification in restored wetlands

Wetlands perform important ecosystem functions, including improvement of water quality through the process of denitrification. To offset the negative environmental impact of replacing wetlands with agriculture and development, the United States has a policy requiring that losses in wetland area are compensated for through wetland restoration elsewhere. However, these restored wetlands may require decades to achieve functional equivalency to natural wetlands. We evaluated the efficacy of using carbon amendments during restoration to promote denitrification potential in four restored wetlands in central New York State, United States. The amendments were straw, topsoil, and biochar, chosen to range along a gradient of carbon lability. Soil samples collected 6 years after restoration were analyzed for denitrification potential and associated soil properties, including soil carbon and nitrogen, pH, microbial biomass carbon and nitrogen, carbon lability, and potential net nitrogen mineralization and nitrification. Compared to unamended control plots, denitrification potential was approximately 3 times higher in straw‐amended plots, 8 times higher in topsoil‐amended plots, and 11 times higher in biochar‐amended plots. Denitrification potential positively correlated with both soil organic carbon and microbial biomass nitrogen, suggesting that the use of soil amendments in restorations can help stimulate the development of denitrification potential by facilitating the suite of carbon and nitrogen cycling processes that underlie this function. However, denitrification potential in a nearby natural reference wetland was at least 50 times higher than it was in the restored wetland plots, highlighting the limitations of using wetland restoration to compensate for the loss of natural wetlands.     

Anaerobic naphthalene degradation by microbial pure cultures under nitrate-reducing conditions

Pure bacterial cultures were isolated from a highly enriched denitrifying consortium previously shown to anaerobically biodegrade naphthalene. The isolates were screened for the ability to grow anaerobically in liquid culture with naphthalene as the sole source of carbon and energy in the presence of nitrate. Three naphthalene-degrading pure cultures were obtained, designated NAP-3-1, NAP-3-2, and NAP-4. Isolate NAP-3-1 tested positive for denitrification using a standard denitrification assay. Neither isolate NAP-3-2 nor isolate NAP-4 produced gas in the assay, but both consumed nitrate and NAP-4 produced significant amounts of nitrite. Isolates NAP-4 and NAP-3-1 transformed 70 to 90% of added naphthalene, and the transformation was nitrate dependent. No significant removal of naphthalene occurred under nitrate-limited conditions or in cell-free controls. Both cultures exhibited partial mineralization of naphthalene, representing 7 to 20% of the initial added (14)C-labeled naphthalene. After 57 days of incubation, the largest fraction of the radiolabel in both cultures was recovered in the cell mass (30 to 50%), with minor amounts recovered as unknown soluble metabolites. Nitrate consumption, along with the results from the (14)C radiolabel study, are consistent with the oxidation of naphthalene coupled to denitrification for NAP-3-1 and nitrate reduction to nitrite for NAP-4. Phylogenetic analyses based on 16S ribosomal DNA sequences of NAP-3-1 showed that it was closely related to Pseudomonas stutzeri and that NAP-4 was closely related to Vibrio pelagius. This is the first report we know of that demonstrates nitrate-dependent anaerobic degradation and mineralization of naphthalene by pure cultures.     

The influence of nitrate on the phosphorus flux to and from oxygen depleted lake sediments

Addition of nitrate to an oxygen depleted sediment leads to a stimulation of the mineralization process if a major part of the nitrate is dissimilatorily reduced. This may cause an increased release of phosphate from some sediments. Nitrate, however, maintains a high redox potential at the sediment surface and thus prevents a release of iron-bound phosphorus. These two counteracting effects of nitrate addition to sediment-water systems were demonstrated in laboratory experiments. A high supply of nitrate to a phosphorus-saturated sediment caused an increased release of phosphate and ammonium once the nitrate had been used up. However, from other sediments there was no or very little release of phosphate but an increased release of ammonium caused by high nitrate dissimilation, probably due to the very high capacity of these sediments to bind liberated phosphate or due to storage of phosphorus in an increased bacterial biomass. Phosphorus fractionation, before and after the experiments with the sediments, provided information on transfers to, from and within the sediments.     

Community analysis of arbuscular mycorrhizal fungi and bacteria in the maize mycorrhizosphere in a long-term fertilization trial

In this study, we investigated the impact of organic and mineral fertilizers on the community composition of arbuscular mycorrhizal (AM) fungi and bacteria in the mycorrhizosphere of maize in a field experiment established in 1956, in south-east Sweden. Roots and root-associated soil aggregates were sampled four times during the growing season in 2005, in control plots and in plots amended with calcium nitrate, ammonium sulphate, green manure, farmyard manure or sewage sludge. Fungi in roots were identified by cloning and sequencing, and bacteria in soil aggregates were analysed by terminal-restriction fragment length polymorphism, cloning and sequencing. The community composition of AM fungi and bacteria was significantly influenced by the different fertilizers. Changes in microbial community composition were mainly correlated with changes in pH induced by the fertilization regime. However, other factors, including phosphate and soil carbon content, also contributed significantly to these changes. Changes in bacterial community composition and a reduction in bacterial taxon richness throughout the growing season were also manifest. The results of this study highlight the importance and significant effects of the long-term application of different fertilizers on edaphic factors and specific groups of fungi and bacteria playing a key role in arable soils.     

The response of denitrifiers in a sandy loam soil affected by a long-term fertilization to organic carbon and nitrate

The response of denitrifiers to carbon in the form of glucose (Glc-C) and nitrate (NO ₃ ⁻ -N) amendments was studied in four differently fertilized plots of sandy-loam soil. Two basically different characteristics of denitrification activity were determined: (1) potential denitrification measured as nitrous oxide production during 1-d incubation in the presence of acetylene, and (2) denitrifying enzyme activity determined in soil slurries as a N₂O production in the presence of acetylene and chloramphenicol during 1 h of incubation. Potential denitrification was strongly influenced by both Glc-C and NO ₃ ⁻ -N amendments in their various combinations, but was also affected by the fertilization practice. The response of denitrifiers to Glc-C and NO ₃ ⁻ -N was generally lower in unfertilized and surprisingly also in highly fertilized soils than in organically and moderately fertilized soils. Denitrifying enzyme activity was stimulated by the fertilization and was, in contrast to potential denitrification, the highest in highly fertilized soil. The results indicate that although active denitrifiers were present in a highly fertilized soil, their ability to develop under optimal conditions was decreased (being similar to that of denitrifiers in unfertilized soil). This suggests long-term changes in soil microbial community in a highly fertilized soil, presumably connected to changes in soil chemistry caused by fertilization.     

Linkages between organic matter mineralization and denitrification in eight riparian wetlands

Denitrification (N₂ production) and oxygen consumption rates were measured at ambient field nitrate concentrations during summer in sediments from eight wetlands (mixed hardwood swamps, cedar swamps, heath dominated shrub wetland, herbaceous peatland, and a wetland lacking live vegetation) and two streams. The study sites included wetlands in undisturbed watersheds and in watersheds with considerable agricultural and/or sewage treatment effluent input. Denitrification rates measured in intact cores of water-saturated sediment ranged from 20 to 260 mol N m^-2 h^-1 among the three undisturbed wetlands and were less variable (180 to 260 mol N M^-2 h^-1) among the four disturbed wetlands. Denitrification rates increased when nitrate concentrations in the overlying water were increased experimentally (1 up to 770 M), indicating that nitrate was an important factor controlling denitrification rates. However, rates of nitrate uptake from the overlying water were not a good predictor of denitrification rates because nitrification in the sediments also supplied nitrate for denitrification. Regardless of the dominant vegetation, pH, or degree of disturbance, denitrification rates were best correlated with sediment oxygen consumption rates (r ² = 0.912) indicating a relationship between denitrification and organic matter mineralization and/or sediment nitrification rates. Rates of denitrification in the wetland sediments were similar to those in adjacent stream sediments. Rates of denitrification in these wetlands were within the range of rates previously reported for water-saturated wetland sediments and flooded soils using whole core¹⁵N techniques that quantify coupled nitrification/denitrification, and were higher than rates reported from aerobic (non-saturated) wetland sediments using acetylene block methods.