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Comparison of combined and separated biological aerated filter (BAF) performance for pre-denitrification/nitrification of municipal wastewater.

The performance of two systems of semi-industrial up-flow biological aerated filters (BAF) with pre-denitrification followed by nitrification was studied and compared under various operating and loading conditions. The first system consisted of two separate reactors for the denitrification and the nitrification step, whereas in the second system the aerobic nitrification zone was packed on top of the anoxic denitrification zone in one reactor. The second system potentially offers substantial savings in investment costs and space requirements for a large scale treatment plant. Regarding the elimination of carbonaceous pollution and denitrification the systems did not show significant differences. However, nitrification in the combined system suffered from the mixing of different biocenosis by daily backwashing and was reduced to 50-70% of the separated system's performance. Factors such as oxygen concentration, raw water composition and loading rates affected both systems' nitrification rates in similar ways. Since it is impossible to optimise the nitrification and denitrification processes separately, the combined system should only be considered for large scale applications if space is very scarce and if a stable raw water composition can be expected. If strict limit values for nitrate have to be met in the effluent, a combination of pre- and post-denitrification is advantageous and advisable.

Bacteria, Aerobic↗

Nitrification-denitrification via nitrite for nitrogen removal from high nitrogen soybean wastewater with on-line fuzzy control.

This study is to demonstrate the performance of a new technology for complete nitrogen removal from soybean wastewater with high nitrogen concentrations. Nitrification-denitrification via nitrite was performed in three kinds of operating condition, i.e. nitrification-denitrification via nitrite under traditional SBR process, nitrification-denitrification via nitrite by alternating oxic/anoxic under fixed-time control and nitrification-denitrification via nitrite by alternating oxic/anoxic under on-line fuzzy control. The best one is nitrification-denitrification via nitrite by alternating oxic/anoxic under on-line fuzzy control. It not only improves both nitrification and denitrification rates and decreases total reaction time, but also saves the amount of added carbon source and alkalinity. In addition, reaction time can be shortened and operation cost can be saved by using real-time fuzzy control aeration and mixing time. So this method enhances the efficiency and the stability of nitrogen removal, and reduces operating costs and construction investment in the process of wastewater treatment.

Agriculture↗

Chemical denitrification for nitrogen removal from landfill leachate.

A new system that removes nitrogen from landfill leachate and other waste waters with similar properties has been proposed with nitritation (i.e. oxidation of ammonium to nitrite) of half of the influent ammonium followed by chemical denitrification with a reaction between equal amounts of ammonium and nitrite to form nitrogen gas. Chemical denitrification occurs at high concentrations and the reactions were studied in combination with a concentration step. Studied concentration methods were freezing/thawing and evaporation/drying. Chemical denitrification is well-known in inorganic chemistry and has been observed in natural systems. Studies in laboratory were focused on chemical denitrification and showed that nearly complete removal of soluble nitrogen can be obtained in evaporation/drying of water solutions or leachate with equal amounts of ammonium and nitrite. Freezing/drying was less efficient with a removal of about 50-60% at high initial concentrations. Chemical denitrification is much influenced by concentration, pH-value, temperature and some compounds in leachate have an inhibiting effect on the reaction. Factors as safety (ammonium nitrite as a salt is explosive above 60 degrees C) and possible side-reactions as formation of ammonia and nitrogen oxides must be carefully evaluated before use in full-scale. Conductivity is a suitable parameter to follow-up the chemical denitrification process.

Ammonia↗

Denitrification of high concentrations of nitrites and nitrates in synthetic medium with different sources of organic carbon. III. Methanol.

The denitrification of nitrites and nitrates (1000 mg N/l) in medium containing methanol as a source of organic carbon was studied. Continuous cultures of mixed population of autochtonic microflora from bottom sludge of nitrogenous wastewater reservoir were set up in a chemostat-type column and packed bed reactor. The efficiency of denitrification of nitrates in packed bed reactor was 506.7 mg N/l/h whereas denitrification of nitrites was from 8.7 to 16.0 mg N/l/h depending on the granulation of the filing material. In the latter case 83% nitrogen was removed from the medium. One of the factors causing low efficiency of denitrification of nitrites is excessive alkalization of the medium in the bed. The use of a three-step bed with adjusted pH resulted in complete denitrification of nitrites with efficiency 60 mg N/l/h. The bacteria inside the bed were dominated by Paracoccus denitrificans and by Pseudomonas aeruginosa when nitrates were present. The sensitivity of P. denitrificans to high concentrations of nitrites seems to be the second factor contributing to low efficiency of denitrification with methanol as organic substrate.

Culture Media↗

Building biofilms for saline hydrogenotrophic denitrification from contrasting origins: Convergent acclimation, divergent performance.

Hydrogenotrophic denitrification is promising for deep nitrogen removal from saline, low-C/N wastewaters, but rapid establishment of stable biofilms at high salinity remains challenging. Here, two saline-adapted inocula from two representative, functionally contrasting habitats-a functionally-diversified inoculum from mangrove sediment and a functionally-focused inoculum from seabed sediment-were acclimated in parallel H2-based membrane biofilm reactors at constant 3.5% salinity. The Diverse-derived biofilm required 80 d to reach steady state and achieved only partial denitrification with 61.1% nitrate removal and considerable nitrite accumulation. In contrast, the Focus-derived biofilm rapidly established complete denitrification within ∼40 d, which was maintained for >50 d, with effluent NOx- below 1 mg-N·L-1 and 98.7% nitrate removal. Microbiome analyses showed that identical operation promoted convergence in community structure and enriched similar community-level functional potentials. However, genome-resolved analysis revealed distinct source-dependent functional organization among dominant microbial populations. Complete denitrifiers co-encoding denitrifying, hydrogenotrophic, and autotrophic functions were preferentially enriched in the Focus-derived biofilm, whereas these functions remained partitioned among different dominant populations in the Diverse-derived biofilm, coinciding with less complete denitrification. These findings indicate that saline hydrogenotrophic denitrification performance depends not only on which functions are enriched at the community level, but also on how key functions become organized among microbial populations, providing a previously overlooked criterion for inoculum selection in saline biological nitrogen control.

Complete denitrification↗

Rhamnolipid production by Pseudomonas aeruginosa under denitrification: effects of limiting nutrients and carbon substrates.

Being biosurfactants, rhamnolipids create severe foaming when produced in aerobic Pseudomonas aeruginosa fermentation. The necessary reduction of aeration causes oxygen limitation and restricts cell and product concentrations. In this study, we evaluate the new strategy of rhamnolipid production under denitrification conditions. Because hydrocarbons used in earlier aerobic fermentations were not metabolizable in the absence of oxygen, other potential C substrates were examined, including palmitic acid, stearic acid, oleic acid, linoleic acid, glycerol, vegetable oil, and glucose. All were found able to support cell growth under anaerobic denitrification. The growth on the two solid substrates (palmitic acid and stearic acid) was slower but could be enhanced substantially by initial addition of rhamnolipids (0.06 g/L). The effects of different limiting nutrients (N, P, S, Mg, Ca, and Fe) were also investigated. The commonly used N limitation could not be adopted in the denitrifying fermentation because the nitrate added for anaerobic respiration would also be assimilated for growth. P limitation was most effective, giving four- to fivefold higher specific productivity than the conventional N limitation. S limitation was comparable to N limitation; Mg limitation was much poorer. Ca and Fe were ineffective in limiting cell growth. The new strategy was further evaluated in a P-limited fermentation with palmitic acid as the substrate. The fermentation was first carried out under denitrification and later switched to aerobic condition. The specific productivity under denitrification was found to be about one-third that of the aerobic condition. The denitrification process was, however, free of foaming or respiratory limitation. Much higher cell concentrations may be employed to attain higher volumetric productivity and product concentrations, for more economical product recovery and/or purification.

Aerobiosis↗

Simultaneous nitrification, denitrification, and phosphorus removal in a lab-scale sequencing batch reactor.

Simultaneous nitrification and denitrification (SND) via the nitrite pathway and anaerobic-anoxic-enhanced biological phosphorus removal (EBPR) are two processes that can significantly reduce the energy and COD demand for nitrogen and phosphorus removal. The combination of these two processes has the potential of achieving simultaneous nitrogen and phosphorus removal with a minimal requirement for COD. A lab-scale sequencing batch reactor (SBR) was operated in alternating anaerobic-aerobic mode with a low dissolved oxygen (DO) concentration (0.5 mg/L) during the aerobic period, and was demonstrated to accomplish nitrification, denitrification, and phosphorus removal. Under anaerobic conditions, COD was taken up and converted to polyhydroxyalkanoates (PHAs), accompanied by phosphorus release. In the subsequent aerobic stage, PHA was oxidized and phosphorus was taken up to <0.5 mg/L by the end of the cycle. Ammonia was also oxidized during the aerobic period, but without accumulation of nitrite or nitrate in the system, indicating the occurrence of simultaneous nitrification and denitrification. However, off-gas analysis showed that the final denitrification product was mainly nitrous oxide (N(2)O), not N(2). Further experimental results demonstrated that nitrogen removal was via nitrite, not nitrate. These experiments also showed that denitrifying glycogen-accumulating organisms (DGAOs), rather than denitrifying polyphosphate-accumulating organisms (DPAOs), were responsible for the denitrification activity.

Acetic Acid↗

Evolution of bacterial denitrification and denitrifier diversity.

Little is known about the role of nitrate in evolution of bacterial energy-generating mechanisms. Denitrifying bacteria are commonly regarded to have evolved from nitrate-respiring bacteria. Some researchers regard denitrification to be the precursor of aerobic respiration; others feel the opposite is true. Currently recognized denitrifying bacteria such as Hyphomicrobium, Paracoccus, Pseudomonas and Thiobacillus form a very diverse group. However, inadequate testing procedures and uncertain taxonomic identification of many isolates may have overstated the number of genera with species capable of denitrification. Nitrate reductases are structurally similar among denitrifying bacteria, but distinct from the enzymes in other nitrate-reducing organisms. Denitryfying bacteria have one of two types of nitrite reductase, either a copper-containing enzyme or an enzyme containing a cytochrome cd moiety. Both types are distinct from other nitrate reductases. Organisms capable of dissimilatory nitrate reduction are widely distributed among eubacterial groups defined by 16S ribosomal RNA phylogeny. Indeed, nitrate reduction is an almost universal property of actinomycetes and enteric organisms. However, denitrification is restricted to genera within the purple photosynthetic group. Denitrification within the genus Pseudomonas is distributed in accordance with DNA and RNA homology complexes. Denitrifiers seem to have evolved from a common ancestor within the purple photosynthetic bacterial group, but not from a nitrate-reducing organism such as those found today. Although denitrification seems to have arisen at the same time as aerobic respiration, the evolutionary relationship between the two cannot be determined at this time.

Bacteria↗

Oxygen requirement for denitrification by the fungus Fusarium oxysporum.

The effects of dioxygen (O2) on the denitrification activity of the fungus Fusarium oxysporum MT-811 in fed-batch culture in a stirred jar fermentor were examined. The results revealed that fungal denitrifying activity requires a minimal amount of O2 for induction, which is repressed by excess O2. The optimal O2 supply differed between the denitrification substrates : 690 micromol O2 x h(-1) (g dry cell wt.)(-1) for nitrate (NO3-) and about 250 micromol O2 x h(-1) (g dry cell wt.)(-1) for nitrite (NO2-). The reduction of NO3- required more O2 than that of NO2- . With an optimal O2 supply, 80% and 52% of nitrogen atoms in NO3- and NO2-, respectively, were recovered as the denitrification product N2O. These features of F. oxysporum differ from those of bacterial denitrifiers that work exclusively under anoxic conditions. The denitrification activity of F. oxysporum MT-811 mutants with impaired NO3- assimilation was about double that of the wild-type strain, suggesting competition for the substrate between assimilatory and dissimilatory types of NO3- reduction. These results showed that denitrification by F. oxysporum has unique features, namely, a minimal O2 requirement and competition with assimilatory NO3-.

Aerobiosis↗

Use of pyrite for pH control during hydrogenotrophic denitrification using metallic iron as the ultimate electron donor.

Maintenance of stable pH through provision of adequate buffering is of importance to many pollutant removal processes where either acid or base is produced as a reaction product. The objective of this study was to evaluate the suitability of pyrite (FeS2) as an in situ buffering agent for arresting pH increase during metallic iron assisted hydrogenotrophic denitrification. Pyrite is considered promising for this purpose because it is a mineral which is unstable under moderately reducing, i.e., anoxic conditions, where such denitrification takes place, and therefore expected to consume hydroxide ions produced due to hydrogenotrophic denitrification reactions and get oxidized to ferrous hydroxide Fe(OH)2. The theoretical basis for this buffering action was established through chemical speciation studies using the chemical speciation software, MINEQL+. Experimental evaluation of the buffering efficiency of pyrite showed that it was effective in arresting pH increase associated with denitrification in both batch systems and during flow through reactive porous media. Further, addition of pyrite had no demonstrable toxic effect on the denitrifying microorganisms, though elevated sulfate concentration was seen in the effluent after denitrification.

Ammonia↗

Electrochemical effect on denitrification in different microenvironments around anodes and cathodes.

A bio-anode reactor and a bio-cathode reactor were developed to investigate the microenvironments around anodes and cathodes and their effects on denitrification. With an applied current of 40 mA, the oxidation-reduction potentials (ORPs) in the bio-cathode and bio-anode reactors were 100-200 mV lower and 50 mV higher, respectively, than that in the control reactor (a normal bio-reactor). The cathode reaction enhanced denitrification and the anode reaction inhibited denitrification. At 40 mA, the denitrification rate in the bio-cathode reactor was 55.1% higher than that in the control reactor. At 75 mA, the denitrification rate in the bio-anode reactor was just 33.5% of that in control reactor. Electric current of less than 20 mA had no effect on the most probable number (MPN) of denitrifiers, but at 75 mA, the MPN of denitrifiers decreased by 90% in the bio-anode reactor. In the bio-cathode reactor, the MPN of denitrifiers increased more than 100% for the lower ORP environment produced by a cathode reaction at 75 mA.

Bacteria↗

Analysis of denitrification genes and comparison of nosZ, cnorB and 16S rDNA from culturable denitrifying bacteria in potato cropping systems.

Bacterial denitrification in agricultural soils is a major source of nitrous oxide, a potent greenhouse gas. This study examined the culturable bacterial population of denitrifiers in arable field soils in potato (Solanum tuberosum L.) production and denitrification genes (nir, nor and nos) and 16S rDNA in those isolates. Enrichments for culturable denitrifiers yielded 31 diverse isolates that were then analysed for denitrification genes. The nitrous oxide reductase (nosZ) gene was found in all isolates. The majority of isolates ( approximately 90%) contained the cnorB nitric oxide reductase gene, with the remainder containing the qnorB gene. Nitrite reductase genes (nirS and nirK) were amplifiable from most of the isolates, and were segregated between species similar to previously isolated denitrifiers. Isolated strains were preliminarily identified using fatty acid methyl ester analysis and further identified using 16S rDNA sequencing. The majority of isolates (21) were classified as Pseudomonas sp., with smaller groups of isolates being most similar to Bosea spp. (4), Achromobacter spp. (4) and two isolates closely related to Sinorhizobium/Ensifer spp. Phylogenetic trees were compared among nosZ, cnorB and 16S rDNA genes for a subset of Pseudomonas strains. The trees were mostly congruent, but some Pseudomonas sp. isolates grouped differently depending on the gene analysed, indicating potential horizontal gene transfer of denitrification genes. Although Bosea spp. are known denitrifiers, to the best of our knowledge this is the first report of isolation and sequencing of denitrification genes from this bacterial genus.

Bacteria↗

Seawater denitrification in a closed mesocosm by a submerged moving bed biofilm reactor.

The performance of a submerged moving bed biofilm reactor (MBBR) for the denitrification of seawater in a 3.25 million closed circuit mesocosm was investigated at pilot scale, using methanol as a carbon source at various C/N ratios. Nitrate accumulation in closed systems where water changes are expensive and problematic may cause toxicity problems to marine life. Seawater was pretreated in a recirculated fixed bed to remove oxygen prior to the denitrification step. The 110l MBBR was partly filled (25%) with spherical positively buoyant polyethylene carriers with an effective surface area of approximately 100 m2 m(-3), which represents 35% of the total surface area. Carriers were maintained submerged by a conical grid and circulated by the downflow jet of an eductor. The MBBR mixing system was designed to prevent dead mixing zones and carrier fouling to avoid sulfate reduction while treating seawater containing as high as 2150 mg SO4-Sl(-1). NO3-N reduction from 53 to as low as 1.7+/-0.7 mg l(-1) and a maximum denitrification rate of 17.7+/-1.4 g Nm(-2) d(-1) were achieved at 4.2-4.3 applied COD/N (w/w) ratio. Methanol consumption corresponded to denitrification stoichiometric values, indicating the absence of sulfate reduction. Denitrification rates and effluent residual dissolved organic carbon were proportional to the C/N ratio. Such reactors could be scaled up in closed systems where water changes must be minimized.

Biofilms↗

Design optimization of a self-cleaning moving-bed bioreactor for seawater denitrification.

The hydrodynamic behavior and denitrification capacity of a down flow self-cleaning moving bed bioreactor (MBBR) using floating packed-bed carriers were investigated. Water mixing in the MBBR was determined by tracer studies, which showed a completely stirred tank reactor. Eleven different types of patented plastic packed-bed carriers were selected based on their density, shape and specific surface area. Video studies of the carriers were used to determine operating conditions and select the Cascade 1A and Tri-Pack 1 carriers for denitrification experiments. Both carriers showed a similar performance level in terms of the denitrification capacity of the resulting MBBR. The improvement in liquid circulation is thought to explain the high denitrification rates that reached 27 gNm(-2)d(-1) and the maintenance of a thin biofilm. Increasing the overall liquid velocity profile led to an increase of up to 30% in the denitrification rate in conditions with a 1-month-old biofilm. This MBBR design seemed to control biofilm development and could easily be scaled up to denitrify seawater or freshwater systems.

Biofilms↗

Kinetic model of autotrophic denitrification in sulphur packed-bed reactors.

Autotrophic denitrification of synthetic wastewater by Thiobacillus denitrificans in upflow sulphur packed-bed reactors was studied in order to establish the process kinetics for prediction of effluent concentration. Elemental sulphur particles of different size served as energy substrate as well as the physical support for the microbial biofilm. Experiments were performed under operating conditions of (i) different flow rates at constant influent nitrate concentration; and (ii) different influent nitrate concentrations at constant flow rate. The experimental results show that autotrophic denitrification rates in upflow sulphur packed-bed reactors can be described by a half-order kinetic model for biofilms. It was found that the half-order kinetic constants of upflow packed-bed reactors are 2.94-3.60, 1.47-2.04, and 1.12-1.29 mg1/2/L1/2 h for sulphur particle sizes of 2.8-5.6, 5.6-11.2, and 11.2-16 mm, respectively. The half-order kinetic constants could be related to the specific surface area of the reactor media by a simple equation. Successful application of the half-order reaction rate model was demonstrated for an actual wastewater (nitrified leachate). A comparison with the literature showed that the half-order reaction rate constants for autotrophic denitrification using elemental sulphur are approximately one order of magnitude lower than those of heterotrophic denitrification. An improved stoichiometric equation for autotrophic denitrification using elemental sulphur as electronic donor is also proposed.

Biodegradation, Environmental↗

Investigation of the denitrification activity of immobilized Pseudomonas butanovora cells in the presence of different organic substrates.

An investigation was made of the effects of variation of the hydraulic retention time (HRT) and the concentrations of three different carbon sources (succinic acid, ethanol and acetic acid) on the denitrification activity of immobilized Pseudomonas butanovora cells. The highest denitrification activity was in all cases measured at a C: N ratio of 6: and a relatively low HRT (2.5 h). The highest denitrification rates were 1.17 kg NO3--N m(-3) d(-1) for succinic acid, 1.63 kg NO3--Nm(-3) d(-1) for ethanol and 1.53kg NO3--Nm(-3) d(-1) for acetic acid. At the same C:N ratios, ethanol and acetic acid proved to be better substrates for the reduction of nitrate and nitrite. The determined optimum C: N ratios were 1.78 +/- 0.31, 0.95 +/- 0.17, 1.76 +/- 0.42 for succinic acid, ethanol and acetic acid, respectively. The optimum C:N ratios for the different substrates did not change in response to an increased flow rate. At a C: N ratio of 3:1 and a HRT of 1.5 h, the immobilized cells did not retain their activity. Apart from the difference in the effectivity between the electron donors, the main influence on the denitrification rate was exerted by the flow rate The results of this study demonstrated that Pseudomonas butanovora can utilize all three of these carbon sources to achieve a high rate of denitrification.

Acetic Acid↗

Combined bioelectrochemical and sulfur autotrophic denitrification for drinking water treatment.

A combined bioelectrochemical and sulfur autotrophic denitrification process for drinking water treatment was put forward and investigated extensively in this paper. In this new process, the bioelectrochemical denitrification was carried out in the upper part of the reactor while sulfur denitrification in the lower part. The H+ produced in Sulfur Part could be consumed by hydrogen denitrification in Bioelectrochemical Part. Therefore, the limestone for pH adjustment in Sulfur Part was not necessary in this combined process, which avoided the problem of hardness increase. The sulfate accumulation in this combined reactor was less than that of the sulfur limestone autotrophic denitrification system. The effluent from two parts was kept neutral at optimum operation conditions. When the influent nitrate was 30 mg-N/L, the reactor could be operated efficiently at the hydraulic retention time ranged from 1.9 to 5h (corresponding minimum current was 16-3 mA), i.e. the effluent NO3(-)-N removal ranged from 90% to 100% without nitrite accumulation and the effluent sulfate concentration was lower than 170 mg/L. The maximum volume-loading rate of the reactor was 0.381 kg NO3(-)-N/(m3d). The biomass and scanning electron microscope micrographs of Sulfur Part were also analyzed.

Bacteria↗

Kinetics of reductive denitrification by nanoscale zero-valent iron.

Zero-valent iron powder (Fe0) has been determined to be potentially useful for the removal of nitrate in the water environment. This research is aimed at subjecting the kinetics of denitrification by nanoscale Fe0 to an analysis of factors affecting the chemical denitrification of nitrate. Nanoscale iron particles with a diameter in the range of 1-100 nm, which are characterized by the large BET specific surface area to mass ratio (31.4 m2/g), removed mostly 50, 100, 200, and 400 mg/l of nitrate within a period of 30 min with little intermediates. Compared with microscale (75-150 microm) Fe0, end product is not ammonia but N2 gas. Kinetics analysis from batch studies revealed that the denitrification reaction with nanoscale Fe0 appeared to be a pseudo first-order with respect to substrate and the observed reaction rate constant (k(obs)) varied with iron content at a relatively low degree of application. The effects of mixing intensity (rpm) on the denitrification rate suggest that the denitrification appears to be coupled with oxidative dissolution of iron through a largely mass transport-limited surface reaction (<40 rpm).

Iron Compounds↗