Effect of bioturbation on denitrification in a marine sediment from the West Mediterranean littoral

Thein vitro effect ofNereis diversicolor on denitrification has been studied in PVC tubes filled with a coastal marine sediment defaunated by sieving. The first aim of the experiment was to determine the effect of sediment defaunation on denitrification (denitrifying population and Denitrifying Enzyme Assays). Sieving induced a loss of 70% of the initial DEA. The number of denitrifying bacteria was 10 times lower than inin situ sediment. In the top two centimetres, the DEA rose by 75% of its initial value, after 82 days. Polychaetes were only added after a return to near pre-disturbance levels to ensure that our data on the effects of their addition would not be disturbed by changes in the sediment. Introduction of Polychaetes increased the denitrifying population and DEA in the first layer (0–2 cm) of the sediment after 15 days. After 45 days, the surface of the polychaete burrows in sediment was 1.3 to 1.5 times higher than after 15 days, resulting in an increase in solute exchange between seawater and the top layer of sediment. An inhibitory effect of oxygen on denitrification was detected in the uppermost layer only.     

Similar temperature responses suggest future climate warming will not alter partitioning between denitrification and anammox in temperate marine sediments

Removal of biologically available nitrogen (N) by the microbially mediated processes denitrification and anaerobic ammonium oxidation (anammox) affects ecosystem N availability. Although few studies have examined temperature responses of denitrification and anammox, previous work suggests that denitrification could become more important than anammox in response to climate warming. To test this hypothesis, we determined whether temperature responses of denitrification and anammox differed in shelf and estuarine sediments from coastal Rhode Island over a seasonal cycle. The influence of temperature and organic C availability was further assessed in a 12‐week laboratory microcosm experiment. Temperature responses, as characterized by thermal optima (T_opt) and apparent activation energy (E_a), were determined by measuring potential rates of denitrification and anammox at 31 discrete temperatures ranging from 3 to 59 °C. With a few exceptions, T_opt and E_a of denitrification and anammox did not differ in Rhode Island sediments over the seasonal cycle. In microcosm sediments, E_a was somewhat lower for anammox compared to denitrification across all treatments. However, T_opt did not differ between processes, and neither E_a nor T_opt changed with warming or carbon addition. Thus, the two processes behaved similarly in terms of temperature responses, and these responses were not influenced by warming. This led us to reject the hypothesis that anammox is more cold‐adapted than denitrification in our study system. Overall, our study suggests that temperature responses of both processes can be accurately modeled for temperate regions in the future using a single set of parameters, which are likely not to change over the next century as a result of predicted climate warming. We further conclude that climate warming will not directly alter the partitioning of N flow through anammox and denitrification.     

Simultaneous occurrence of denitrification and nitrate ammonification in sediments of the French Mediterranean Coast

Dissimilatory nitrate reductions in coastal marine sediment of Carteau Cove (French Mediterranean Coast) were studied between April 1993 and July 1994. Simultaneous determination of denitrification and dissimilatory nitrate reduction to ammonium was achieved by using a combination of acetylene blockage and 15N techniques. After short incubations (maximum 5 h), a part of 15N labelled nitrate added to the sediment was recovered as ammonium without incorporation in organic matter. The result indicate that a fraction of nitrate was reduced to ammonium by a dissimilatory mechanism instead of denitrifying. Denitrifying and nitrate ammonifying activities ranged from 0 to 19.8 μmol l-1 d-1 and from 2.3 to 83.2 μmol l-1 d-1, respectively. Denitrification rates were highest in early spring whereas nitrate ammonification were highest in fall. The recovery of nitrate reduced as N2O-N plus ammonium was between 40 and 100%, the highest nitrogen losses were recorded in July. Depending on the station and time of year denitrification accounted for between 0 and 43% of the total nitrate reduction whereas dissimilatory nitrate reduction to ammonium (DNRA) accounted for between 18 and 100%. The reduction rate data suggest that the pathway of nitrate reduction to ammonium may be important in coastal sediments. This revised version was published online in July 2006 with corrections to the Cover Date.     

The denitrification potential and density of denitrifying bacteria in the bottom sediments of Matupi Harbor (Papua New Guinea)

The denitrification potential and density of denitrifying bacteria were determined in suboxidized sediments of Matupi Harbor (Papua New Guinea). The sediments were characterized by low denitrification potentials, not exceeding 0.8 nM/(g h). The maximum density of denitrifying bacteria was recorded in the upper sediment horizons and was 10⁴ cells/g of wet ground.     

Methanogenesis,sulfate reduction and crude oil biodegradation in hot Alaskan oilfields

Petrochemical and geological evidence suggest that petroleum in most reservoirs is anaerobically biodegraded to some extent. However, the conditions for this metabolism and the cultivation of the requisite microorganisms are rarely established. Here, we report on microbial hydrocarbon metabolism in two distinct oilfields on the North Slope of Alaska (designated Fields A and B). Signature anaerobic hydrocarbon metabolites were detected in produced water from the two oilfields offering evidence of in situ biodegradation activity. Rate measurements revealed that sulfate reduction was an important electron accepting process in Field A (6–807 µmol S l^?1 day^?1), but of lesser consequence in Field B (0.1–10 µmol S l^?1 day^?1). Correspondingly, enrichments established at 55°C with a variety of hydrocarbon mixtures showed relatively high sulfate consumption but low methane production in Field A incubations, whereas the opposite was true of the Field B enrichments. Repeated transfer of a Field B enrichment showed ongoing methane production in the presence of crude oil that correlated with ≥ 50% depletion of several component hydrocarbons. Molecular‐based microbial community analysis of the methanogenic oil‐utilizing consortium revealed five bacterial taxa affiliating with the orders Thermotogales, Synergistales, Deferribacterales (two taxa) and Thermoanaerobacterales that have known fermentative or syntrophic capability and one methanogen that is most closely affiliated with uncultured clones in the H₂‐using family Methanobacteriaceae. The findings demonstrate that oilfield‐associated microbial assemblages can metabolize crude oil under the thermophilic and anaerobic conditions prevalent in many petroleum reservoirs.     

Bioturbation and the role of microniches for sulfate reduction in coastal marine sediments

The effects of bioturbation in marine sediments are mainly associated with an increase in oxic and oxidized zones through an influx of oxygen‐rich water deeper into the sediment and the rapid transport of particles between oxic and anoxic conditions. However, macrofaunal activity also can increase the occurrence of reduced microniches and anaerobic processes, such as sulfate reduction. Our goal was to determine the two‐dimensional distribution of microniches associated with burrows of a ghost shrimp (Neotrypaea californiensis) and to determine microbial activities. In laboratory experiments, detailed measurements of sulfate reduction rates (SRR) were measured by injecting, in a 1 cm grid, radiolabelled sulfate directly into a narrow aquarium (40 cm × 30 cm × 3 cm) containing the complex burrow of an actively burrowing shrimp. Light‐coloured oxidized burrow walls, along with black reduced microniches, were clearly visible through the aquarium walls. Direct injection of radiotracers allowed for whole‐aquarium incubation to obtain two‐dimensional documentation of sulfate reduction. Results indicated SRR were up to three orders of magnitude higher (140–790 nmol SO₄^2? cm^?3 day^?1) in reduced microniches associated with burrows when compared with the surrounding sediment. Additionally, some of the subsurface sulfate‐reducing microniches associated with the burrow system appeared to be zones of dinitrogen fixation. Bioturbation may also lead to decreased sulfate reduction in other microniches and the sum of the activity in all microniches might not result in a total increase of sulfate reduction compared with non‐bioturbated control sediments.     

Effects of specific inhibitors on anammox and denitrification in marine sediments

The effects of three metabolic inhibitors (acetylene, methanol, and allylthiourea [ATU]) on the pathways of N2 production were investigated by using short anoxic incubations of marine sediment with a 15N isotope technique. Acetylene inhibited ammonium oxidation through the anammox pathway as the oxidation rate decreased exponentially with increasing acetylene concentration; the rate decay constant was 0.10+/-0.02 microM-1, and there was 95% inhibition at approximately 30 microM. Nitrous oxide reduction, the final step of denitrification, was not sensitive to acetylene concentrations below 10 microM. However, nitrous oxide reduction was inhibited by higher concentrations, and the sensitivity was approximately one-half the sensitivity of anammox (decay constant, 0.049+/-0.004 microM-1; 95% inhibition at approximately 70 microM). Methanol specifically inhibited anammox with a decay constant of 0.79+/-0.12 mM-1, and thus 3 to 4 mM methanol was required for nearly complete inhibition. This level of methanol stimulated denitrification by approximately 50%. ATU did not have marked effects on the rates of anammox and denitrification. The profile of inhibitor effects on anammox agreed with the results of studies of the process in wastewater bioreactors, which confirmed the similarity between the anammox bacteria in bioreactors and natural environments. Acetylene and methanol can be used to separate anammox and denitrification, but the effects of these compounds on nitrification limits their use in studies of these processes in systems where nitrification is an important source of nitrate. The observed differential effects of acetylene and methanol on anammox and denitrification support our current understanding of the two main pathways of N2 production in marine sediments and the use of 15N isotope methods for their quantification.     

Potential denitrification in submerged natural and impacted sediments of Lake Batata,an Amazonian lake

Lake Batata is one of many clear water lakes located on the floodplain of the Trombetas River in the northern Brazilian Amazon. Lake Batata is distinctly different from other lakes of this region because, for a period of 10 years, its waters received tremendous amounts of aluminum ore tailings from a bauxite mining operation. Approximately 30% of the sediments of the upper basin of the 2100 hectare lake were covered by tailings before dumping was curtailed. The goal of this research was to identify factors controlling denitrification in the natural and impacted sediments of Lake Batata. Rates of denitrification in sediments were estimated in the laboratory by the acetylene blockage method. Denitrification was measured under four conditions: without amendment; amended with glucose; amended with nitrate; and amended with glucose and nitrate. Denitrification was observed only in assays amended with nitrate suggesting that availability of nitrate is a principle factor for controlling denitrification in the sediments of Lake Batata. Effects of nitrate amendments are most pronounced when the water level is low, i.e. during the hydroperiods of draw-down and low-water.     

Measurement of bacterial sulfate reduction in sediments: Evaluation of a single-step chromium reduction method

A procedure which includes the Total Reduced Inorganic Sulfur (TRIS) in a single distillation step is described for the radiotracer measurement of sulfate reduction in sediments. The TRIS includes both Acid Volatile Sulfide (AVS: H₂S + FeS) and the remaining Chromium Reducible Sulfur (CRS: S⁰, FeS₂). The single-step distillation was simpler and faster than the consecutive distillations of AVS and CRS. It also resulted in higher (4–50%) sulfate reduction rates than those obtained from the sum of³⁵S in AVS and CRS. The difference was largest when the sediment had been dried after AVS but before CRS distillation. Relative to the³⁵S-AVS distillation alone, the³⁵S-TRIS single-step distallation yielded 8–87% higher reduction rates. The separation and recovery of FeS, S⁰ and FeS₂ was studied under three distillation conditions: 1) cold acid, 2) cold acid with Cr²⁺, and 3) hot acid with Cr²⁺. The FeS was recovered by cold acid alone while pyrite was recovered by cold acid with Cr²⁺. A smaller S⁰ fraction, presumably of the finer crystal sizes, was recovered also in the cold acid with Cr²⁺ while most of the S⁰ required hot acid with Cr²⁺ for reduction to H₂S.     

Major substrates for microbial sulfate reduction in the sediments of Ise Bay, Japan

To clarify the anaerobic microbial interactions in the process of carbon mineralization in marine eutrophic environments, the microbial sulfate reduction and methane production rates were examined in coastal marine sediments of Ise Bay, Japan, in autumn 1990. Sulfate reduction rates (51–210 nmol ml⁻¹ day⁻¹ at 24°C) were much higher than the methane production ones (<1.78 nmol ml⁻¹ day⁻¹) in the surface sediments (top 2 cm) at the six stations surveyed (water depth: 10.7–23.3 m). Substrates for sulfate-reducing bacteria (SRB) were estimated after the addition of a specific inhibitor for SRB (20 mmol l⁻¹ molybdate) into the sediment slurry, from the substrate accumulation rates. In the presence of the inhibitor, sulfate reduction was completely stopped and volatile fatty acids (mainly acetate) were accumulated, although hydrogen was not. Methane production occurred markedly accompanied by consumption of the accumulated acetate from the third day after the addition of molybdate. The maximum rate of methane production was 1.2–1.9 μmol ml⁻¹ day⁻¹, which was similar to those in highly polluted freshwater sediments such as the Tama River, Tokyo, Japan. These results show that acetate is a common major substrate for sulfate reduction and methane production, and SRB competitively inhibit potential acetoclastic methanogenesis in coastal sediments. Methanogens may potentially inhabit the sediments at low levels of population density and activity.     

Competition between nitrate and nitrite reduction in denitrification by Pseudomonas fluorescens

A pure culture of Pseudomonas fluorescens was used as a model system to study the kinetics of denitrification. An exponentially growing culture was harvested and resuspended in an anoxic acetate solution buffered with K/Na phosphate at pH values of 6.6, 7.0, 7.4, and 7.8. The temperature was kept at 28 degrees C in all assays. Nitrate pulses of approximately 0.2 mg N/L caused nitrite to accumulate due to a faster rate of nitrate reduction over nitrite reduction. The rate of nitrate reduction was observed to depend on its concentration as predicted by the Michaelis-Menten equation. At nonlimiting nitrate concentrations, nitrite reduction was described by the same equation. Otherwise, nitrite reduction also depended on nitrate concentration. Consequently, nitrate and nitrite reductions compete with each other for the oxidation of common electron donors. A kinetic model for nitrate competitive inhibition of nitrite reduction is proposed. The model was used to interpret the nitrate and nitrite profiles observed at the four pH values: the optimum pH value was 7.0 in both cases; the affinity for nitrite was also not affected by the medium pH in the range of values 6.6 to 7.4 (K(mNO(3) ) = 0.04 mg N/L); the affinity for nitrite was also not affected by the medium pH in the range of values 6.6 to 7.4 (K(mNO(2) ) = 0.06 mg N/L), but it decreased sharply for the pH value of 7.8. Although the ratio between the two maximum reduction rates (V(max NO(2) )/V(max NO(3) )) is constant, nitrite accumulation depends on the medium pH value. Therefore, the regulation mechanism that shifts the electron flow between the two terminal reductases is readily reversible and does not change their relative maximum reduction rates. (c) 1995 John Wiley & Sons, Inc.     

Assessment of heavy metal pollution and ecological risk in marine sediments (A case study: Persian Gulf)

This study was made to determine the pollution status and potential ecological risk of heavy metals in sediment of Persian Gulf. Surface sediments were collected seasonally by Peterson grab, and the concentrations of heavy metals were measured by using inductively coupled plasma–optical emission spectrometry (ICP–OES). The range concentrations obtained in mg/kg were 10,800–22,400 for Fe, 5.32–10.12 for Pb, 24.63–42.38 for Ni, 22.52–39.46 for Cu, and 31.64–47.20 for Cr. The concentrations of Pb, Ni, Cu, and Cr have been found lower than the Interim Sediment Quality Guidelines and probable effect level values suggesting that heavy metal contents in sediments from area of study would not be expected to cause adverse biological effects on the biota. The obtained enrichment factor values for various metals were between minimal enrichment (Pb = 0.5) and extremely enrichment (Cu = 3.11). The values of I_geo for Pb, Ni, Cu, and Cr were characterized under no pollution (0). The highest value of potential ecological risk index (RI) (8.36) was observed at St. 4 while the lowest value (5.25) was detected at station 6. Based on potential ecological RI, the Persian Gulf had low ecological risk.     

Effect of the cathode potential and sulfate ions on nitrate reduction in a microbial electrochemical denitrification system

Recently, bioelectrochemical systems have been demonstrated as advantageous for denitrification. Here, we investigated the nitrate reduction rate and bacterial community on cathodes at different cathode potentials [?300, ?500, ?700, and ?900 mV vs. standard hydrogen electrode (SHE)] in a two-chamber microbial electrochemical denitrification system and effects of sulfate, a common nitrate co-contaminant, on denitrification efficiency. The results indicated that the highest nitrate reduction rates (3.5 mg L^?1 days^?1) were obtained at a cathode potential of ?700 mV, regardless of sulfate presence, while a lower rate was observed at a more negative cathode potential (?900 mV). Notably, although sulfate ions generally inhibited nitrate reduction, this effect was absent at a cathode potential of ?700 mV. Polymerase chain reaction–denaturing gradient gel electrophoresis revealed that bacterial communities on the graphite-felt cathode were significantly affected by the cathode potential change and sulfate presence. Shinella-like and Alicycliphilus-like bacterial species were exclusively observed on cathodes in reactors without sulfate. Ochrobactrum-like and Sinorhizobium-like bacterial species, which persisted at different cathode potentials irrespective of sulfate presence, were shown to contribute to bioelectrochemical denitrification. This study suggested that a cathode potential of around ?700 mV versus SHE would ensure optimal nitrate reduction rate and counteract inhibitory effects of sulfate. Additionally, sulfate presence considerably affects denitrification efficiency and microbial community of microbial electrochemical denitrification systems.     

Anaerobic ammonium oxidation linked to sulfate and ferric iron reduction fuels nitrogen loss in marine sediments

Availability of fixed nitrogen is a pivotal driver on primary productivity in the oceans, thus the identification of key processes triggering nitrogen losses from these ecosystems is of major importance as they affect ecosystems function and consequently global biogeochemical cycles. Denitrification and anaerobic ammonium oxidation coupled to nitrite reduction (Anammox) are the only identified marine sinks for fixed nitrogen. The present study provides evidence indicating that anaerobic ammonium oxidation coupled to the reduction of sulfate, the most abundant electron acceptor present in the oceans, prevails in marine sediments. Tracer analysis with ¹⁵N-ammonium revealed that this microbial process, here introduced as Sulfammox, accounts for up to 5 μg ¹⁵N₂ produced g^?1 day^?1 in sediments collected from the eastern tropical North Pacific coast. Raman and X-ray diffraction spectroscopies revealed that elemental sulfur and sphalerite (ZnFeS) were produced, besides free sulfide, during the course of Sulfammox. Anaerobic ammonium oxidation linked to Fe(III) reduction (Feammox) was also observed in the same marine sediments accounting for up to 2 μg ¹⁵N₂ produced g^?1 day^?1. Taxonomic characterization, based on 16S rRNA gene sequencing, of marine sediments performing the Sulfammox and Feammox processes revealed the microbial members potentially involved. These novel nitrogen sinks may significantly fuel nitrogen loss in marine environments. These findings suggest that the interconnections among the oceanic biogeochemical cycles of N, S and Fe are much more complex than previously considered.     

Sulfte-reduction process in sediments of Lake Kinneret,Israel

Monomictic Lake Kinneret is stratified during summer and autumn, resulting in a hypolimnion rich in H₂S (3–7 mg 1^–1). In winter and spring every year a bloom of dinoflagallate Peridinium gatunense produces an average biomass of 150000 ton wet weight. Part of this biomass sinks to the hypolimnion and sediments where it is decomposed and mineralized, with some of the mineralization due to the activity of sulfate-reducing bacteria (SRB). The sulfate-reduction potential of the upper sediment layer at the deepest part of the lake (42 m) was measured. The activity of the enzyme arylsulfatase was also monitored. Rates of sulfate-reduction ranged from a minimum of 12 nmoles SO_f4 ^p2–-reduced cm^–3 day^–1 in December before lake overturn to a maximum of 1673 nmoles SO_f4 ^p2– reduced cm^–3 day^–1 in July during stratification. These rates are considerably higher than those recorded from other freshwater lakes in the world and are probably limited more by the availability of organic matter than by sulfate concentrations.     

不同类型原油污染土壤生物修复技术研究

对不同类型原油污染土壤在实用规模的预制床上采用堆制技术进行生物修复 .通过投加肥料、菌剂、控制水分和pH ,可使微生物获得较好的生态环境 .当稀油、高凝油、特稠油和稠油污染的土壤中原油总量为 2 5 .8～ 77.2 g·kg-1土时 ,经过近 2个月的运行 ,石油总量的去除率可达 38.37%～ 5 6 .74 % .石油中芳烃、沥青和胶质混合物是制约石油快速降解的主要因素 .在处理过程中筛选出石油降解的优势菌株 ,其中有 6株真菌、6株细菌和 1株放线菌 .研究结果为石油污染土壤异位生物修复技术实用化提供了理论依据 .