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PubMed1996

The effect of antibiotics on nitrification processes. Batch assays.

The effect of different antibiotics at several concentrations of ampicillin (0-250 mg/L), benzylpenicillin (0-250 mg/L), novobiocine (0-150 mg/L), oxytetracycline (0-250 mg/L), and chloramphenicol (0-50 mg/L) on a stabilized nitrifying sludge was evaluated under aerated and lithoautotrophic conditions. No effect resulting from the presence of antibiotics on the biomass and nitrate production was noticed. The specific growth rate and volumetric nitrification rate average values for the controls were 8.28 x 10-3/h-1 and 2.74 x 10-3 g/L.h, respectively. Similar rate values were found when different kinds of antibiotic and concentrations were tested. These results may be explained by the nature of the floc or the instability of the antibiotics.

Aerobiosis↗
PubMed2001

Inhibition of aerobic growth and nitrification of bacteria in sewage sludge by antibacterial agents.

Toxicity of antibacterial agents on environmentally relevant bacteria was investigated using activated sludge. The growth and nitrifying inhibiting effects for activated sludge of benzyl penicillin (penicillin G) (BP), tetracycline (TC), chlortetracycline (CTC), oxytetracycline (OTC), olaquindox (O), streptomycin (ST), tiamulin (TI), tylosin (TYL) sulfadiazine (SDZ), metronidazole (MET), and oxolinic acid (OXA) was investigated. Studies were performed in accordance to the ISO 15522 (1999) and ISO 9509 (1989) test guidelines, respectively. The toxicity (EC50 value, mg/L) found with the ISO 15522 was in decreasing EC50 values; O (95.7), BP (84.6), TYL (54.7), TI (14.3), TC (2.2), OTC (1.2), ST (0.47), CTC (0.40), and OXA (0.1). No observed effect concentrations (NOECs) (mg/L) of 100 and 60, respectively was found for MET and SDZ. Triplicate tests assessing the effects of the antibacterial agents on the nitrification rate gave indications only as the level of increased or decreased rate. More accurate data for the inhibition of Nitrosomonas europaea was found with a suspended culture of the nitrifying bacteria. The toxicity (EC50 value, mg/L) found was in decreasing EC50 values; TI (23.3), SDZ (17.0), TC (4.0), OTC (1.7), OXA (1.0), CTC (0.64), O (0.03), ST (0.02). For MET and TYL, NOECs (mg/L) of 100 and 50 were found, respectively. The antibacterial agents were also assessed using a pour plate method with both (separately tested) activated sludge bacteria and N. europeae showing to be 5 to 10 times more potent to most agents except SDZ, TI, and MET.

Ammonia↗
PubMed2006

Nitrogen removal in a wastewater treatment plant through biofilters: nitrous oxide emissions during nitrification and denitrification.

In order to estimate N(2)O emissions from immersed biofilters during nitrogen removal in tertiary treatments at urban wastewater treatment plants (WWTPs), a fixed culture from the WWTP of "Seine Centre" (Paris conurbation) was subjected to lab-scale batch experiments under various conditions of oxygenation and a gradient of methanol addition. The results show that during nitrification, N(2)O emissions are positively related to oxygenation (R (2) = 0.99). However, compared to the rates of ammonium oxidation, the percentage of emitted N(2)O is greater when oxygenation is low (0.5-1 mgO(2) L(-1)), representing up to 1% of the oxidized ammonium (0.4% on average). During denitrification, the N(2)O emission reaches a significant peak when the quantity of methanol allows denitrification of between 66% and 88%. When methanol concentrations lead to a denitrification of close to 100%, the flows of N(2)O are much lower and represent on average 0.2% of the reduced nitrate. By considering these results, we can estimate, the emissions of N(2)O during nitrogen removal, at the "Seine Centre" WWTP, to approximately 38 kgN-N(2)O day(-1).

Bioreactors↗
PubMed2007

Short-cut nitrification of domestic wastewater in a pilot-scale A/O nitrogen removal plant.

The feasibility of nitrite accumulation in a pilot-scale A/O (anoxic/oxic) nitrogen removal plant treating domestic wastewater was investigated at various dissolved oxygen (DO) concentrations and pH levels. The results showed that the pH was not a useful operational parameter to realize nitrite accumulation. Significant nitrite accumulation was observed at the low DO concentration range of 0.3-0.8 mg/l and the maximum nitrite accumulation ratio of about 90% occurred at a DO concentration of 0.6 mg/l. This suggests a reduction of 22% in the oxygen consumption, and therefore a considerable saving in aeration. However, the nitrite accumulation was destroyed at the high DO concentration and the resumption was very slow. In addition, the average ammonia removal efficiency reached as high as 93% at the low DO level. Moreover, experimental results indicated that nitrogen could be removed by simultaneous nitrification and denitrification (SND) via nitrite in the aerobic zones at the low DO concentration, with the efficiency of 6-12%.

Bacteria, Aerobic↗
PubMed2006

Molecular analysis of the nitrogen cycle in deep-sea microorganisms from the Nankai Trough: genes for nitrification and denitrification from deep-sea environmental DNA.

Nitrification and denitrification are bacterial functions, which are important for the global nitrogen cycle. Thus, it is important to study the diversity and distribution of bacteria in the environment, which are involved in the nitrogen cycle on the earth. Ammonia monooxygenase encoded by the amoA gene and nitrite reductase encoded by nirK or nirS are essential enzymes for nitrificaton and denitrification, respectively. These genes can be used as markers for the identification of organisms in the nitrogen cycle. In this study, we identified amoA (42 clones) and nirS (98 clones) genes in parallel from samples recovered from the deep-sea of the Nankai Trough. Genes for nirK could not be amplified from these samples. The obtained amoA sequences were not so closely related to those of amoA genes from previously isolated environmental organisms and those of genes from environmental DNAs. On the other hand, the nirS genes sequenced showed some relationship to some extent with the latter genes. However, some of the newly sequenced genes formed clusters, which contained no previously identified genes on a phylogenetic tree. These are likely present in specific denitrifiers from the deep-sea. The results of this study further suggest that nitrifiers and denitrifiers live in the same area of the Nankai Trough and the nitrogen cycle exists even in the deep-sea.

DNA↗
PubMed2005

Nitrification and denitrification in high-strength ammonium by Alcaligenes faecalis.

Alcaligenes faecalis sp. No. 4, that has the ability of heterotrophic nitrification and aerobic denitrification in high-strength ammonium at about 1200 mg-N/l, converted about one-half of removed NH4+-N to intracellular nitrogen and nitrified only 3% of the removed NH4+. From the nitrogen balance, 40-50% of removed NH4+-N was estimated to be denitrified. Production of N2 was confirmed by GC-MS and 90% of denitrified products was N2. The maximum ammonium removal rate, 29 mg-N/l h and its denitrification rate in aerated batch experiments, were 5-40 times higher than those of other bacteria with the same ability.

Aerobiosis↗
PubMed2006

Kinetic analysis of the inhibitory effect of trichloroethylene (TCE) on nitrification in cometabolic degradation.

In this study, the inhibitory effect of TCE on nitrification process was investigated with an enriched nitrifier culture. TCE was found to be a competitive inhibitor of ammonia oxidation and the inhibition constant (K(I)) was determined as 666-802 microg/l. The TCE affinity for the AMO enzyme was significantly higher than ammonium. The effect of TCE on ammonium utilization was evaluated with linearized plots of Monod equation (e.g., Lineweaver-Burk, Hanes-Woolf and Eadie-Hofstee plots) and non-linear least square regression (NLSR). No significant differences were found among these data evaluation methods in terms of kinetic parameters obtained.

Ammonia↗
PubMed1988

Experimental work related to nitrogen deposition, nitrification and soil acidification--a case study.

The leaching of major ions has been studied since August 1986 in two plots with different nitrogen fertilization levels and in a control plot in a 29-year-old stand of Norway spruce (Picea abies Karst.) in south-central Sweden. The fertilization started in 1967. The two fertilizer levels, both of which have caused a significant stimulation of the tree growth, correspond to an annual input of approximately 35 kg N ha(-1) and 75 kg N ha(-1) respectively, as NH4NO3. Phosphorus fertilizer is also applied. Field and laboratory incubations performed during 1986 showed that nitrification mainly occurs in the plot with the highest fertilization level, in accordance with the fact that nitrate could not be detected in the soil water in the other two plots. Fertilization has increased the ionic strength of the soil solution due mainly to sulphate in the phosphate fertilizer, but also nitrate at the highest fertilization level. This has caused an increase in total aluminium and a decline in pH. The preliminary data reported here are compared with results obtained at Swedish field sites with moderate to high levels of nitrogen deposition.

Journal Article↗
PubMed1990

An examination of proton translocation and energy conservation during heterotrophic nitrification.

Whether selected heterotrophic nitrifiers, as do the autotrophs, conserve energy during the oxidation of their nitrogenous substrates was studied. The examination of proton translocation of four different bacterial nitrifiers capable of pyruvic oxime [(PO), CH3-C(NOH)-COOH] nitrification and by an NH4+ oxidizing Arthrobacter sp. was initiated. Three of the PO nitrifying bacteria, all pseudomonads, oxidize hydroxylamine (NH2OH) at a greater rate than PO and yielded only stoichiometric protons when NH2OH was the reductant. The fourth bacterium, Alcaligenes faecalis ATCC 8750, an adept PO oxidizer, does not appreciably oxidize NH2OH. The bacterium displayed----H+NH2OH ratios far less than if NH2OH was stoichiometrically converted to nitrite. When given NH4+, the Arthrobacter sp. yielded proton translocation patterns which were inconsistent with the metabolic data collected concerning NH4+ oxidation. Thus no data was collected which supported energy conservation via proton translocation by these heterotrophic nitrifiers.

Alcaligenes↗
PubMed2005

Aerobic nitrification-denitrification by heterotrophic Bacillus strains.

Twenty-four Bacillus strains predominantly outgrown in a night soil treatment system were isolated and characterized. Under various culture conditions, cell interactions took place among them and cell population changed. Maximum removal of NH4+-N and cell production by the isolates occurred under the conditions of 30% DO and C/N ratio of 8. Five dominant isolates were identified to be species of Bacillus cereus, Bacillus subtilis and Bacillus licheniformis with similarities of 78-94%. Additions of 0.8% peptone and 0.3% yeast extract to a basal medium influenced the growth of isolates and the removal of NH4+-N in flask culture. Metal ions such as Ca2+, Fe2+ and Mg2+ had a similar effect. The specific growth rates of the five isolates were found to be in a range of 0.43-0.55 h(-1). During the flask experiment of nitrogen removal under aerobic growth conditions, active nitrification by the isolates occurred largely in 1h with a decrease of COD and alkalinity reduced to only 74.6% of theoretical value. From the nitrogen balance, the percentage of nitrogen lost in the flask culture was estimated to be 33.0%, which was presumed to convert to N2 gas. This conversion of ammonia to N2 without formation of nitrous oxide under aerobic growth conditions was confirmed by GC analysis. From all the results, it has been found that the Bacillus strains were able to occur simultaneously aerobic nitrification/denitrification and the B3 process using the Bacillus strains seemed to possess some economic advantages.

Aerobiosis↗
PubMed2005

Simultaneous nitrification and formaldehyde biodegradation in an activated sludge unit.

The simultaneous removal of formaldehyde and ammonium in a lab-scale activated sludge unit was investigated. The unit was operated at a hydraulic retention time of 2.4 days with an ammonium concentration in the influent of 350 mg NH4+-N/L, maintaining the ammonium loading rate at 0.15 g NH4+-N/Ld during the operation time. However, the applied organic loading rate was increased stepwise by increasing the formaldehyde concentration from 26 up to 3168 mg/L, corresponding to 0.01-1.40 g COD/Ld. High formaldehyde removal efficiencies, around 99.5% (+/-0.38), were maintained at all the formaldehyde concentrations. Ammonium removal was also very high during the operation period, around 99.9% (+/-0.01). The ammonium concentration in the effluent was lower than 0.1 mg NH4+-N/L at all applied organic loading rates, indicating that there was no inhibition of nitrification by formaldehyde.

Ammonia↗
PubMed2005

Mitigating nitrous oxide and methane emissions from soil in rice-wheat system of the Indo-Gangetic plain with nitrification and urease inhibitors.

Mitigation of methane (CH4) and nitrous oxide (N2O) emissions from soil is important to reduce the global warming. Efficacy of five nitrification inhibitors, i.e. neem (Azadirachta melia) cake, thiosulphate, coated calcium carbide, neem oil coated urea and dicyandiamide (DCD) and one urease inhibitor, hydroquinone, in mitigating N2O and CH4 emissions from fertilized soil was tested in rice-wheat system in the Indo-Gangetic plains. The closed chamber technique was used for the collection of gas samples, which were analyzed using gas chromatography. Reduction in N2O emission on the application of nitrification/urease inhibitors along with urea ranged from 5% with hydroquinone to 31% with thiosulphate in rice and 7% with hydroquinone to 29% with DCD in wheat crop. The inhibitors also influenced the emission of CH4. While application of neem coated urea, coated calcium carbide, neem oil and DCD reduced the emission of CH4; hydroquinone and thiosulphate increased the emission when compared to urea alone. However, the global warming potential was lower with the inhibitors (except hydroquinone) as compared to urea alone, suggesting that these substances could be used for mitigating greenhouse gas emission from the rice-wheat systems.

Acetylene↗
PubMed2005

Identifying causes for N2O accumulation in a lab-scale sequencing batch reactor performing simultaneous nitrification, denitrification and phosphorus removal.

The recently described process of simultaneous nitrification, denitrification and phosphorus removal (SNDPR) has a great potential to save capital and operating costs for wastewater treatment plants. However, the presence of glycogen-accumulating organisms (GAOs) and the accumulation of nitrous oxide (N(2)O) can severely compromise the advantages of this process. In this study, these two issues were investigated using a lab-scale sequencing batch reactor performing SNDPR over a 5-month period. The reactor was highly enriched in polyphosphate-accumulating organisms (PAOs) and GAOs representing around 70% of the total microbial community. PAOs were the dominant population at all times and their abundance increased, while GAOs population decreased over the study period. Anoxic batch tests demonstrated that GAOs rather than denitrifying PAOs were responsible for denitrification. N(2)O accumulated from denitrification and more than half of the nitrogen supplied in a reactor cycle was released into the atmosphere as N(2)O. After mixing SNDPR sludge with other denitrifying sludge, N(2)O present in the bulk liquid was reduced immediately if external carbon was added. We therefore suggest that the N(2)O accumulation observed in the SNDPR reactor is an artefact of the low microbial diversity facilitated by the use of synthetic wastewater with only a single carbon source.

Bacteria, Aerobic↗
PubMed2007

Dynamics of changes in methanogenesis and associated microflora in a flooded alluvial soil following repeated application of dicyandiamide, a nitrification inhibitor.

Influence of repeated application of the nitrification inhibitor dicyandiamide (DCD), on CH(4) production and associated microflora in a flooded alluvial soil, was investigated in a laboratory incubation study. Application of DCD at the time of soil incubation resulted in a substantial reduction in CH(4) production (31% over that of untreated control). Second repeat application of DCD, on the contrary, annulled the inhibitory effect on CH(4) production, restoring it to the level of unamended soil. Application of the third dose of DCD maintained CH(4) production almost to the same extent as that of second application. The alleviation of the initial inhibitory effect of DCD on CH(4) production was linked to the enhanced degradation of DCD following its repeated application to the flooded soil. Admittedly, abatement of the initial inhibitory effect of DCD on CH(4) production in soil repeatedly amended with DCD was also related to the inhibition of CH(4)-oxidizing bacterial population and noticeable stimulation of heterotrophic bacterial population. Results suggest that repeat application of DCD with fertilizer-N to flooded rice soils might not be effective in controlling CH(4) production under field condition.

Anaerobiosis↗
PubMed2006

Why is metabolic labour divided in nitrification?

Winogradsky discovered in 1890 that nitrification is carried out in two consecutive steps by two distinct groups of bacteria: ammonia-oxidizing bacteria and nitrite-oxidizing bacteria. An explanation for this division of labour is offered based on the kinetic theory of optimal design of metabolic pathways, which postulates the existence of an optimal length for a pathway that maximizes the rate of ATP production. Shortening long pathways could, therefore, increase growth rate. However, this would reduce growth yield if the shorter pathway has fewer ATP-generating steps. High yields would be advantageous when bacteria grow in clonal clusters, as is typical for biofilms. It is postulated that bacteria that completely oxidize ammonia to nitrate exist in such environments.

Ammonia↗
PubMed2006

Simultaneous nitrification and p-cresol oxidation in a nitrifying sequencing batch reactor.

The tolerance, kinetic behavior and oxidizing ability of a nitrifying sludge exposed to different initial concentrations of p-cresol (25-150mg/l) were evaluated in a sequencing batch reactor (SBR) fed with 200mg NH(4)(+)-N/ld. The nitrifying SBR operated up to 300mg/ld of p-cresol, achieving simultaneously the complete ammonium oxidation to nitrate and the total consumption of p-cresol and its transitory intermediates from the culture. p-Cresol induced a significant decrease in the values for specific rates of ammonium consumption, showing that the ammonium oxidation pathway was mainly inhibited. After 7 months of operation in SBR, the specific rates of NH(4)(+)-N oxidation, NO(3)(-)-N formation, and total organic carbon consumption were 0.6g NH(4)(+)-N/g microbial protein-Nh, 0.3g NO(3)(-)-N/g microbial protein-Nh, and 0.24g total organic carbon/g microbial protein h, respectively. The microbial growth rate was always low (maximum value of 12.2+/-0.4mg protein-N/ld) and settleability of the sludge was good with sludge volume index values lower than 21ml/g. The oxidation of p-cresol and its intermediates was carried out faster throughout the cycles and nitrification inhibition decreased with the number of cycles.

Bioreactors↗
PubMed2001

Nitrification and denitrification as a source for NO and NO2 production in high-strength wastewater.

Laboratory and half-technical scale experiments were performed to evaluate nitric oxide (NO) and nitrogen dioxide (NO2) production during biological N-elimination from wastewater with high ammonium concentration (about 700 mg N L-1). In a laboratory scale bioreactor with biomass retention, the ammonia oxidizer Nitrosomonas europaea and the denitrifier Paracoccus denitrificans were grown as reference organisms in co-culture in order to simulate the nitrifying and denitrifying community of wastewater treatment plants. Synthetic wastewater and sludge liquor from the municipal wastewater treatment plant in Lueneburg (Germany) were used. In the laboratory scale reactor, during the treatment of synthetic wastewater, 0.28% of the oxidized ammonium-N was released as NO-N by a pure culture of Nitrosomonas. A simultaneously nitrifying and denitrifying co-culture only released 0.04 to 0.2%. NO2 formation was not observed. NO production was much higher in sludge liquor. A pure culture of Nitrosomonas produced 0.52% NO + NO2-N (= NOx-N), a co-culture of Nitrosomonas and Paracoccus even 1.64% NOx-N. The production rate strongly depended on the media and the organisms used. In a co-culture of N. europaea and P denitrificans, Nitrosomonas was shown to be the most efficient NO producer. NO production increased with ammonium oxidation rate and with nitrite concentration of the medium. In synthetic wastewater, NO production was not influenced by reduced oxygen content. However, in sludge liquor NO production rate increased with decreasing O2 concentration. Here, for the first time, the formation of significant amounts of NO2 during simultaneous nitrification/denitrification could be demonstrated. In half-technical scale experiments, only 0.07% of the oxidized ammonium-N was released as NO-N from the nitrification stage. NO2 was not detectable. Release of nitric oxide from the denitrification stage was mainly diffusion limited and the amount produced did not exceed 0.0001%. A calculation on the basis of the results presented, revealed that biological treatment of nitrogen-rich wastewater is not a significant source for pollution of the atmosphere with NOx in industrial areas.

Biodegradation, Environmental↗
PubMed2002

Model-based evaluation of temperature and inflow variations on a partial nitrification-ANAMMOX biofilm process.

A mathematical model describing nitrification (nitritification plus nitratification) and anaerobic ammonium oxidation (ANAMMOX) combined in a biofilm reactor was developed. Based on this model, a previously proposed one-reactor completely autotrophic ammonium removal over nitrite (CANON) process was evaluated for its temperature dependency and behaviour under variable inflow. The temperature-dependency of growth rates of the involved organisms is described by an Arrhenius-type equation. If temperature decreases, the activities of the involved organisms decrease. This means that thicker biofilms are needed or the ammonium surface load (ASL) to the biofilm should be decreased to maintain full N-removal at lower temperatures. Although the growth rate of nitrite oxidisers is higher than that of ammonium oxidisers at lower temperatures, these organisms can be effectively competed out due to a lower oxygen affinity. Variable inflow or dissolved oxygen (DO) concentration negatively affect the N-removal efficiency due to an unbalance between applied ASL load and required oxygen concentration. A variation of the dissolved oxygen concentration in a small range (+/- 0.2g O2/m3) has no significant influence on the process performance, which means that requirements on electrode sensitivity and a DO control scheme are not too stringent. A variable ASL has obvious influence on the process performance, at both constant and variable DO. A good adjustment of DO in accordance with the variable ASL is needed to optimise the N-removal efficiency. At T = 20 degrees C, an N-removal efficiency of 88% is possible at ASL = 0.5 g NH4+ - N/mr2 d, in a biofilm of at least 0.7 mm thickness and a DO level of 0.3 g O2/m3 in the bulk liquid.

Biofilms↗