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PubMed1978

Simultaneous determinations of nitrification and nitrate reduction in coastal sediments by a 15N dilution technique.

Nitrification, the oxidation of ammonia to nitrite and nitrate, and nitrate reduction by bacteria in coastal sediments of Mangoku-Ura and Odawa Bay were simultaneously determined by a 15N dilution technique. In muddy sediments of Mangoku-Ura, nitrate reduction proceeded at a rate of 10(-2) to 10 X 10(-2) microgram-atoms of N/g per h. Nitrification was far less intensive. Denitrification, or N2 production from nitrate, accounted for about 30% of the nitrate reduction. A simultaneous occurrence of nitrification and nitrate reduction with a similar rate of 10(-2) microgram-atoms of N/g per h was demonstrated in sandy sediment collected from a Zostera bed of Odawa Bay.

Ammonia↗
PubMed1982

Effects of polychlorinated biphenyls and environmental biotransformation products on aquatic nitrification.

The effects of polychlorinated biphenyls (PCBs) on nitrification were examined for pure cultures and natural reservoir samples. PCBs at concentrations greater than 10 microgram liter-1 inhibited nitrification, principally ammonium oxidation, in one of two natural reservoir environments. However, this inhibition could not be reproduced in pure high-cell-density cultures or in previously contaminated reservoir waters. A PCB environmental biotransformation product, p-chlorophenylglyoxylic acid, and p-chloromandelic acid had no effect on nitrification.

Biotransformation↗
PubMed1983

Estimates of denitrification and nitrification in coastal and estuarine sediments.

Denitrification and nitrification in sediments of Tama Estuary and Odawa Bay, Japan, were investigated by the combined use of a continuous-flow sediment-water system and a N tracer technique. At Odawa Bay, the nitrification rate was comparable to the nitrate reduction rate, and 70% of the N(2) evolved originated from nitrogenous oxides (nitrate and nitrite) which were produced by the action of nitrifying bacteria in the sediments. At Tama Estuary, the nitrate reduction rate was 11 to 17 times higher than the nitrification rate, and nitrogenous oxides derived from ammonium accounted for only 6 to 9% of the N(2) evolution by denitrification.

Journal Article↗
PubMed1984

Heterotrophic nitrification among denitrifiers.

Twelve denitrifying bacteria representing six genera were tested for an ability to nitrify pyruvic oxime heterotrophically. Six of these bacteria exhibited appreciable nitrification activity, yielding as much as 5.8 mM nitrite and little or no nitrate when grown in a mineral salts medium containing 7 mM pyruvic oxime and 0.05% yeast extract. Of the six active bacteria, four (Pseudomonas denitrificans, Pseudomonas aeruginosa, and two strains of Pseudomonas fluorescens) could grow on yeast extract but not pyruvic oxime, one (Pseudomonas aureofaciens) could grow slowly on pyruvic oxime, and one (Alcaligenes faecalis) could apparently grow on pyruvic oxime in the presence of yeast extract but not in its absence. Eight of the twelve bacteria in the resting state could oxidize hydroxylamine to nitrite, and P. aureofaciens was remarkably active in this regard. In general, those denitrifiers active in the nitrification of pyruvic oxime or hydroxylamine or both are abundant in soils. A possible advantage of having nitrification and denitrification capabilities in the same organism is discussed.

Alcaligenes↗
PubMed1984

Identification of heterotrophic nitrification in a sierran forest soil.

A potential for heterotrophic nitrification was identified in soil from a mature conifer forest and from a clear-cut site. Potential rates of NO(2) production were determined separately from those of NO(3) by using acetylene to block autotrophic NH(4) oxidation and chlorate to block NO(2) oxidation to NO(3) in soil slurries. Rates of NO(2) production were similar in soil from the forest and the clear-cut site and were strongly inhibited by acetylene. The rate of NO(3) production was much greater than that of NO(2) production, and NO(3) production was not significantly affected by acetylene or chlorate. Nitrate production was partially inhibited by cycloheximide, but was not significantly reduced by streptomycin. Neither the addition of ammonium nor the addition of peptone stimulated NO(3) production. N labeling of the NH(4) pool demonstrated that NO(3) was not coming from NH(4). The potential for heterotrophic nitrification in these forest soils was greater than that for autotrophic nitrification.

Journal Article↗
PubMed1989

Effect of fluoride on nitrification of a concentrated industrial waste.

The potential for biological nitrification of an industrial waste containing 4,000 mg of ammonia N (NH(4)-N) and 10,000 mg of fluoride per liter was investigated. Ammonium sulfate and sodium fluoride were tested in various combinations of 100 to 2,000 mg of NH(4)-N per liter and 0 to 5,000 mg of F per liter in suspended-growth stirred-tank reactors containing enriched cultures of nitrifying bacteria from a municipal sewage treatment plant. The stirred-tank reactors were fed once per day at a constant hydraulic retention period and cell retention time of 10 days. Temperature was 23 degrees C, and pH was 7.0 to 7.5. Clarified secondary effluent was used to make up feeds and to provide minor nutrients. Steady-state data, confirmed by mass balances, were obtained after five to six retention periods. In the absence of fluoride, nitrification efficiency was near 100% for up to 500 mg of NH(4)-N per liter. The influence of fluoride was studied at a low ammonia concentration (100 mg/liter) and exerted no significant effect on nitrification at concentrations of up to 200 mg/liter. Maximum effect of fluoride was reached at 800 mg of F per liter, and no greater inhibition was observed for up to 5,000 mg of F per liter. At the highest concentrations studied, ion pairing of ammonium and fluoride may exert a significant effect on kinetic coefficients. Kinetic analyses showed maximum specific substrate removal rates (q(max)) of NH(4)-N to be about 2.3 mg of N per mg of volatile suspended solids per day in the absence of fluoride and 0.85 mg of N per mg of volatile suspended solids per day in the presence of fluoride. The form of inhibition due to the presence of fluoride was shown to be not competitive, conforming to a mixed inhibition model.

Journal Article↗
PubMed1992

Denitrification, dissimilatory reduction of nitrate to ammonium, and nitrification in a bioturbated estuarine sediment as measured with N and microsensor techniques.

Nitrogen and oxygen transformations were studied in a bioturbated (reworked by animals) estuarine sediment (Norsminde Fjord, Denmark) by using a combination of N isotope (NO(3)), specific inhibitor (C(2)H(2)), and microsensor (N(2)O and O(2)) techniques in a continuous-flow core system. The estuarine water was NO(3) rich (125 to 600 muM), and NO(3) was consistently taken up by the sediment on the four occasions studied. Total NO(3) uptake (3.6 to 34.0 mmol of N m day) corresponded closely to N(2) production (denitrification) during the experimental steady state, which indicated that dissimilatory, as well as assimilatory, NO(3) reduction to NH(4) was insignificant. When C(2)H(2) was applied in the flow system, denitrification measured as N(2)O production was often less (58 to 100%) than the NO(3) uptake because of incomplete inhibition of N(2)O reduction. The NO(3) formed by nitrification and not immediately denitrified but released to the overlying water, uncoupled nitrification, was calculated both from NO(3) dilution and from changes in NO(3) uptake before and after C(2)H(2) addition. These two approaches gave similar results, with rates ranging between 0 and 8.1 mmol of N m day on the four occasions. Attempts to measure total nitrification activity by the difference between NH(4) fluxes before and after C(2)H(2) addition failed because of non-steady-state NH(4) fluxes. The vertical distribution of denitrification and oxygen consumption was studied by use of N(2)O and O(2) microelectrodes. The N(2)O profiles measured during the experimental steady state were often irregularly shaped, and the buildup of N(2)O after C(2)H(2) was added was much too fast to be described by a simple diffusion model. Only bioturbation by a dense population of infauna could explain these observations. This was corroborated by the relationship between diffusive and total fluxes, which showed that only 19 to 36 and 29 to 62% of the total O(2) uptake and denitrification, respectively, were due to diffusion-reaction processes at the regular sediment surface, excluding animal burrows.

Journal Article↗
PubMed1995

Differential inhibition by allylsulfide of nitrification and methane oxidation in freshwater sediment.

Addition of nitrapyrin, allylthiourea, C(inf2)H(inf2), and CH(inf3)F to freshwater sediment slurries inhibited CH(inf4) oxidation and nitrification to similar extents. Dicyandiamide and allylsulfide were less inhibitory for CH(inf4) oxidation than for nitrification. Allylsulfide was the most potent inhibitor of nitrification, and the estimated 50% inhibitory concentrations for this process and CH(inf4) oxidation were 0.2 and 121 (mu)M, respectively. At a concentration of 2 (mu)M allylsulfide, growth and CH(inf4) oxidation activity of Methylosinus trichosporium OB3b were not inhibited. Allylsulfide at 200 (mu)M inhibited the growth of M. trichosporium by approximately 50% but did not inhibit CH(inf4) oxidation activity. Nitrite production by cells of M. trichosporium was not significantly affected by allylsulfide, except at a concentration of 2 mM, when growth and CH(inf4) oxidation were also inhibited by about 50%. Methane monooxygenase activity present in soluble fractions of M. trichosporium was not inhibited significantly by allylsulfide at either 200 (mu)M or 2 mM. These results suggest that the partial inhibition of CH(inf4) oxidation in sediment slurries by high allylsulfide concentrations may be caused by an inhibition of the growth of methanotrophs rather than an inhibition of methane monooxygenase activity specifically. We conclude that allylsulfide is a promising tool for the study of interactions of methanotrophs and nitrifiers in N cycling and CH(inf4) turnover in natural systems.

Journal Article↗
PubMed1998

A bioluminescence assay using Nitrosomonas europaea for rapid and sensitive detection of nitrification inhibitors.

An expression vector for the luxAB genes, derived from Vibrio harveyi, was introduced into Nitrosomonas europaea. Although the recombinant strain produced bioluminescence due to the expression of the luxAB genes under normal growing conditions, the intensity of the light emission decreased immediately, in a time-and dose-dependent manner, with the addition of ammonia monooxygenase inhibitors, such as allylthiourea, phenol, and nitrapyrin. When whole cells were challenged with several nitrification inhibitors and toxic compounds, a close relationship was found between the change in the intensity of the light emission and the level of ammonia-oxidizing activity. The response of bioluminescence to the addition of allylthiourea was considerably faster than the change in the ammonia-oxidizing rate, measured as both the O2 uptake and NO2- production rates. The bioluminescence of cells inactivated by ammonia monooxygenase inhibitor was recovered rapidly by the addition of certain substrates for hydroxylamine oxidoreductase. These results suggested that the inhibition of bioluminescence was caused by the immediate decrease of reducing power in the cell due to the inactivation of ammonia monooxygenase, as well as by the destruction of other cellular metabolic pathways. We conclude that the assay system using luminous Nitrosomonas can be applied as a rapid and sensitive detection test for nitrification inhibitors, and it will be used to monitor the nitrification process in wastewater treatment plants.

Enzyme Inhibitors↗
PubMed2005

Autotrophic ammonia-oxidizing bacteria contribute minimally to nitrification in a nitrogen-impacted forested ecosystem.

Deposition rates of atmospheric nitrogenous pollutants to forests in the San Bernardino Mountains range east of Los Angeles, California, are the highest reported in North America. Acidic soils from the west end of the range are N-saturated and have elevated rates of N-mineralization, nitrification, and nitrate leaching. We assessed the impact of this heavy nitrogen load on autotrophic ammonia-oxidizing communities by investigating their composition, abundance, and activity. Analysis of 177 cloned beta-Proteobacteria ammonia oxidizer 16S rRNA genes from highly to moderately N-impacted soils revealed similar levels of species composition; all of the soils supported the previously characterized Nitrosospira clusters 2, 3, and 4. Ammonia oxidizer abundance measured by quantitative PCR was also similar among the soils. However, rates of potential nitrification activity were greater for N-saturated soils than for soils collected from a less impacted site, but autotrophic (i.e., acetylene-sensitive) activity was low in all soils examined. N-saturated soils incubated for 30 days with ammonium accumulated additional soluble ammonium, whereas less-N-impacted soils had a net loss of ammonium. Lastly, nitrite production by cultivated Nitrosospira multiformis, an autotrophic ammonia-oxidizing bacterium adapted to relatively high ammonium concentrations, was significantly inhibited in pH-controlled slurries of sterilized soils amended with ammonium despite the maintenance of optimal ammonia-oxidizing conditions. Together, these results showed that factors other than autotrophic ammonia oxidizers contributed to high nitrification rates in these N-impacted forest soils and, unlike many other environments, differences in nitrogen content and soil pH did not favor particular autotrophic ammonia oxidizer groups.

Ammonia↗
PubMed1971

Nitrification of aspartate by Aspergillus flavus.

Heterotrophic conversion of l-aspartic acid to nitrification products by Aspergillus flavus was studied in a replacement incubation system. Numerous amino acids supported nitrification; aspartate and glutamate were about equivalent as the best sources of nitrate. Addition of sodium bicarbonate to the incubation system substantially enhanced nitrate formation for all nitrifiable amino acids except aspartic acid, but the basis for the bicarbonate effect is obscure. The yield of nitrate from l-aspartate was not approached by forms of aspartic acid resulting from substitution on the beta carbon, the amino nitrogen, or the gamma carboxyl group or by aspartate presented as the d-configuration. There was no relationship between nitrate formation and the occurrence of such possible intermediates as nitrite, bound hydroxylamine, ammonia, aspergillic acid, and beta-nitropropionic acid. Uniformly labeled (14)C-l-aspartate that was nitrified in replacement incubation led to no accumulation of label in possible nitrification products in the culture filtrate. Label was found in components of the mycelium after acid hydrolysis, with heaviest accumulation in what appeared to be glucosamine and an unidentified compound, possibly acetylglucosamine. Detectable label was redistributed into serine, glycine, and threonine.

Amino Acids↗
PubMedDate not supplied

Potential nitrification and denitrification on different surfaces in a constructed treatment wetland.

Improved understanding of the importance of different surfaces in supporting attached nitrifying and denitrifying bacteria is essential if we are to optimize the N removal capacity of treatment wetlands. The aim of this study was therefore to examine the nitrifying and denitrifying capacity of different surfaces in a constructed treatment wetland and to assess the relative importance of these surfaces for overall N removal in the wetland. Intact sediment cores, old pine and spruce twigs, shoots of Eurasian watermilfoil (Myriophyllum spicatum L.), and filamentous macro-algae were collected in July and November 1999 in two basins of the wetland system. One of the basins had been constructed on land that contained lots of wood debris, particularly twigs of coniferous trees. Potential nitrification was measured using the isotope-dilution technique, and potential denitrification was determined using the acetylene-inhibition technique in laboratory microcosm incubations. Nitrification rates were highest on the twigs. These rates were three and 100 times higher than in the sediment and on Eurasian watermilfoil, respectively. Potential denitrification rates were highest in the sediment. These rates were three times higher than on the twigs and 40 times higher than on Eurasian watermilfoil. The distribution of denitrifying bacteria was most likely due to the availability of organic material, with higher denitrification rates in the sediment than on surfaces in the water column. Our results indicate that denitrification, and particularly nitrification, in treatment wetlands could be significantly increased by addition of surfaces such as twigs.

Bacteria↗
PubMedDate not supplied

Pilot-scale evaluation of separate-stage nitrification using an attached-growth, moving-bed media process.

Submerged attached-growth processes, both fixed and moving bed, are becoming more popular. These processes may or may not be used in combination with suspended-growth treatment. The objective of this project was to evaluate a tertiary attached-growth, moving-bed media nitrification system based on the ammonia-nitrogen removal rates and effluent concentrations that could be achieved. A pilot-scale system for nitrification of secondary municipal wastewater effluent was operated for 142 days. The minimum aeration requirements for media mixing provided sufficient dissolved oxygen. The generation of biological solids from nitrification was insignificant. Design effluent ammonia-nitrogen concentrations (<6 mg/L) were achieved at influent loading rates greater than design (1.18 g ammonia-nitrogen/[m2 x d]) coincident with detention times shorter than design conditions (4 hours). Empirical relationships for the ammonia-nitrogen removal rate and effluent concentration as a function of influent ammonia-nitrogen loading were developed. Detention time is believed to be an important parameter affecting these relationships.

Ammonia↗
PubMed1993

Simultaneous nitrification and denitrification using immobilized microorganisms.

Nitrogen removal from wastewaters is a multiple step process in which nitrification is often a problem due to the slow growth rates of the nitrifying bacteria. By immobilization of these bacteria, nitrification can be efficiently accomplished in compact reactors. In this paper, the possibilities of integrated nitrification and denitrification within a single reactor system are evaluated. Two main systems are studied: a) Nitrosamines europaea and Pseudomonas denitrificans co-immobilized in a a gel bead and operated in an air-lift reactor; b) the same bacteria separately immobilized and operated in different compartments of a multiple gas-lift reactor.

Ammonia↗
PubMed2001

Exceptionally high-rate nitrification in sequencing batch reactors treating high ammonia landfill leachate.

The nitrogen removal capacity of a suspended culture system treating mature landfill leachate was investigated. Leachate containing high ammonium levels of 300-900 mg N/L was nitrified in a bench scale sequencing batch reactor. Leachate from four different landfills was treated over a two year period for the removal of nitrogen. In this time, a highly specific nitrifying culture was attained that delivered exceptionally high rates of ammonia removal. No sludge was wasted from the system to increase the throughput and up to 13 g/L of MLSS was obtained. Settleability of the purely nitrifying biomass was excellent with SVI less than 40 mL/g, even at the high sludge concentrations. Nitrification rates up to 246 mg N/(L h) (5.91 g N/(L d)) and specific nitrification rates of 36 mg N/(gVSS h) (880 mg N/(gVSS d)) were obtained. The loading to the system at this time allowed complete nitrification of the leachate with a hydraulic retention time of only 5 hours. Following these successful treatability studies, a full-scale plant was designed and built at one of the landfills investigated.

Ammonia↗
PubMed2001

How partial nitrification could improve reclaimed wastewater transport in long pipes.

Reclaimed wastewater transport is studied in a concrete-lined cast iron pipe, where a nitrification-denitrification process occurs. The pipe is part of the Reuse System of Reclaimed Wastewater of South Tenerife (Spain), 0.6 m in diameter and 61 km long. In order to improve wastewater quality, at 10 km from the inlet there is injection of fresh water saturated in dissolved oxygen (DO), after which a fast nitrification process usually appears (less than two hours of space time). The amount of oxidized nitrogen compounds produced varies between 0.8 and 4.4 mg/l NOx(-)-N. When DO has disappeared, a denitrification process begins. The removal of nitrite is complete at the end of the pipe, whereas the nitrate does not disappear completely, leaving a concentration of about 0.4-0.5 mg/l. For a COD/NOx(-)-N ratio higher than 5, a first order nitrification rate in NOx(-)-N has resulted, with the constant k20 = 0.079 h-1, for a NOx(-)-N concentration range of 0.8-4.4 mg/l. Finally the following temperature dependency for the first order denitrification rate constant has been found: k = k20 x 1 x 15T-20. Although nitrogen could be used as nutrient in the agricultural reuse, its removal from reclaimed wastewater could be useful in order to diminish the chlorine needs for reclaimed wastewater disinfection.

Agriculture↗
PubMed1996

Nitrification and ammonification in aquatic systems.

Nitrification is an essential step in the nitrogen cycle of natural systems because it links organic matter degradation to fixed nitrogen loss. Ammonium released by ammonification is oxidized to nitrate by nitrification, and can then be reduced to dinitrogen gas by denitrification, resulting in net loss of fixed nitrogen from the system. Whether organic matter degradation results in net ammonium release depends largely on the quality of the organic substrate and interactions among members of the microbial community involved in nitrogen and organic matter cycling. In sediments, nitrogen cycle processes depend on the supply of organic matter and oxygen from overlying water. The nature of the net flux (which direction and which form of nitrogen) is a function of closely coupled reactions (ammonification-nitrification-denitrification) in the nitrogen cycle.

Ammonia↗