Modes of action of nitrification inhibitors

 In recent years, substantial progress has been made towards understanding the modes of action for the specific inhibition of autotrophic NH₃ oxidation. This has included demonstrating that NH₃ monooxygenase (AMO) has a broad substrate range for catalytic oxidation, and the inhibitory effects of many compounds are due to competition for the active site. Other compounds, such as acetylenes, are oxidized by the normal catalytic cycle of AMO to highly reactive products which covalently bind the enzyme causing irreversible inhibition. Substantial evidence has shown the important role of Cu in the activity of AMO, and indicated that a large class of compounds containing thiono-S inhibit AMO activity by binding with Cu within the active site. Heterocyclic N compounds form another important class of nitrification inhibitors with little known about their mode of action, although evidence suggests that their inhibitory influence is closely related to the presence of ring N. Received: 21 July 1998     

Some furfural derivatives as nitrification inhibitors

Three series of furfural derivatives, namely N-O-furfural oxime ethers, furfural Schiff bases (furfurylidene anilines), and furfural chalcones, have been synthesized and evaluated for nitrification inhibition activity in laboratory incubation studies in typic Ustocrept soil. Furfural oxime ethers and furfural Schiff bases showed potential activity, but furfural chalcones were only mildly active. N-O-ethyl furfural oxime among the oxime ethers, and furfurylidine-4-chloroaniline among the furfural Schiff bases, performed the best. These two compounds showed more than 50% nitrification inhibition on the 45th day at 5% dose as compared to 73% inhibition by nitrapyrin. Activity of furfural oxime ethers decreased with an increase in carbon atoms in the N-O-alkyl side chain. Introduction of a chlorine atom in the phenyl ring of furfurylidene anilines increased the persistence of their activity. N-O-Ethyl furfural oxime and furfurylidine-4-chloroaniline coated urea performed at par with their application in solution form. Ethyl and N-O-isopropyl oxime, as well as chloro- and nitro- substituted Schiff bases, did not reveal any phytotoxicity (adverse effect on germination) on chickpea seeds (Cicer arietinum) even at the highest dose (40 ppm, soil basis).     

Effect of nitrification inhibitors on rice protein

Abstract

The effects of ammonium sulfate and urea nitrogen (150 kg N/ha) applied with three levels (5, 10 and 15% of N) of the nitrification inhibitors karanjin and nitrapyrin on grain protein of rice (Oryza sativa L. cv. Bala) were studied in pot experiment. Karanjin at the 10 and 15 per cent levels and nitrapyrin at the 10 per cent level significantly increased grain protein. Rice protein levels were highest at the 15 per cent karanjin level.     

Decomposition rate of dicyandiamide and nitrification inhibition

Abstract

Degradation of dicyandiamide (DCD) was assayed in laboratory studies at 8, 15, and 22 C in a Decatur silt loam and in a Norfolk loamy sand. Dicyandiamide was very short lived at 22 C, with half‐lives of 7.4 and 14.7 days in the Decatur and Norfolk soils, respectively. In the Norfolk soil at 8 C, half‐life increased to 52.2 days. In a nitrificaton study of both soils at 22 C, 80 mg (NH₄)₂SO₄‐N kg^‐1 of soil was applied with 20 mg DCD‐N kg^‐1 of soil and 100 mg kg^‐1 (NH₄)₂S0₄‐N was added with 5% nitrapyrin. Distinct lag phases preceded zero order nitrification with the inhibitor treatments. Lag periods were 2 and 2.6 times the half life of DCD in the degradation study for Decatur and Norfolk soils, respectively. Like most nitrification inhibitors, the effectiveness of DCD decreases with increasing temperature. In the Norfolk loamy sand, nitrification inhibition by DCD was equal to nitrapyrin for up to 42 days, but in Decatur silt loam, DCD was less potent to nitrapyrin as a nitrification inhibitor.     

Growth of comammox Nitrospira is inhibited by nitrification inhibitors in agricultural soils

Purpose

The discovery of comammox Nitrospira being capable of complete oxidising ammonia to nitrate radically challenged the conventional concept of two-step nitrification. However, the response of comammox Nitrospira to nitrification inhibitors (NIs) and their role in soil nitrification remain largely unknown, which has hindered our ability to predict the efficiency of NIs in agroecosystems.

Materials and methods

We evaluated the effect of four NIs, 2-chloro-6-(trichloromethyl) pyridine (nitrapyrin), 3,4-dimethylpyrazole phosphate (DMPP), allylthiourea (ATU) and dicyandiamide (DCD) on the growth of comammox Nitrospira, ammonia-oxidising archaea (AOA) and ammonia-oxidising bacteria (AOB) in two pasture and arable soils.

Results and discussion

The amendment of nitrogen fertiliser significantly increased soil nitrate concentrations over time, indicating a sustaining nitrification activity in both soils. The addition of all the four NIs effectively reduced the production of nitrate in both soils, but to varying degrees during incubation. The abundances of comammox Nitrospira clade A were significantly increased by addition of nitrogen fertilisers and significantly impeded by the four NIs in the pasture soil, but their abundances were only remarkably hindered by nitrapyrin in the arable soil. All the four NIs obviously inhibited the AOB abundances in both soils. Except for DMPP, the other three NIs effectively suppressed the AOA abundances in both soils.

Conclusions

We provided new evidence that growth of comammox Nitrospira clade A can be stimulated by nitrogen fertilisers and inhibited by various nitrification inhibitors, suggesting their potential role in nitrification of agricultural soils.

     

Effect of urease inhibitors on nitrification in soil

Abstract. This paper describes experiments on the inhibitory effects of hydroquinone (HQ), phenylphos-phorodiamidate (PPDA) and N-butyl phosphorothioic triamide (NBPT) on the nitrification in soil. Incubations were carried out at 2/3 field capacity at 25°C of soil samples to which either ammonium or nitrite was added together with inhibitors. Addition of PPDA or NBPT did not influence the oxidation of ammonium. HQ. however, retarded the process significantly, and also the accumulation of nitrite. This was confirmed in experiments whereby added nitrite was followed. Some of the differences could be explained by changes in the soil pH. During incubation the evolution of the total mineral nitrogen was not importantly altered by addition of the inhibitors.     

Potentials of nitrification inhibitors in modern N-fertilizer management

Modern agricultural practices require a new concept of N-fertilizer management in order to optimize N-utilization and avoid N-losses. Nitrification inhibitors or ?N-stabilizers”? fit very good into this conception. Dicyandiamide (DCD) is an efficient nitrification inhibitor and blocks the first step of nitrification for 1–3 months (depending on temperature). This effect is bacteriostatic (not bactericidal) and does not affect other (esp. C-heterotrophic) soil microorganisms (?biological activity”?). DCD is a non-toxic, water soluble compound and will be degraded to CO₂, NH₃ and H₂O without any residues. There are various possibilities to use DCD: addition to liquid manure temporarily prevents oxidation of ammonium nitrogen e.g. of slurry or waste water from potato starch production. In combination with inorganic fertilizers like ammonium sulfate or urea (with 10% of total-N) it enables the farmer to control NH₄-supply to crop plants in certain stages of growth and to gain certain operational advantages by less frequently split applications of N especially on sand and rendzina soils. Thus, the systematic use of nitrification inhibitors not only represents a progress in agricultural technique but also helps to substantially reduce risks concerning pollution of surface and ground waters that are sometimes inevitable consequences of agricultural production.     

Nanoclay polymer composites loaded with urea and nitrification inhibitors for controlling nitrification in soil

Nanoclay polymer composites (NCPCs) were synthesized with partially neutralized acrylic acid and bentonites and loaded with urea and nitrification inhibitors (NIs) to act as a slow release carrier of nitrogen (N). The resulting product was characterized by Fourier transform infrared spectroscopy (FTIR), scanning electron microscopy (SEM) and X-ray diffraction (XRD). The XRD of NCPCs revealed that the bentonite layers were completely exfoliated and dispersed in the composite after the polymerization. The water absorbency of pure polymer (Acrylic acid + Acrylamide) was 197.53 g g^?1 and 137.75 g g^?1 by nanocomposite (8% nanobentonite) in distilled water. The nitrification inhibition ability of these NCPCs was evaluated by incubation study for 60 days in laboratory at 28°C and 50% water-holding capacity. The Schiff base -NCPCs were most effective at inhibiting nitrification (30–87%) compared to dicyandiamide and Neem oil. A column study was performed to know the movement of NH₄-N and NO₃-N at three different depths. Result showed that the Schiff base -NCPC decreased nitrate movement by 78.5% at the depth of 5 cm in soil column. The slow release of nitrogen and good water retention capacity confirmed that these NCPCs can be viably exploited for application in agriculture.     

Suppression of methane oxidation in aerobic soil by nitrogen fertilizers,nitrification inhibitors,and urease inhibitors

Concentrations of CH₄, a potent greenhouse gas, have been increasing in the atmosphere at the rate of 1% per year. The objective of these laboratory studies was to measure the effect of different forms of inorganic N and various N-transformation inhibitors on CH₄ oxidation in soil. NH ₄ ⁺ oxidation was also measured in the presence of the inhibitors to determine whether they had differential activity with respect to CH₄ and NH ₄ ⁺ oxidation. The addition of NH₄Cl at 25 g N g^-1 soil strongly inhibited (78–89%) CH₄ oxidation in the surface layer (0–15 cm) of a fine sandy loam and a sandy clay loam (native shortgrass prairie soils). The nitrification inhibitor nitrapyrin (5 g g^-1 soil) inhibited CH₄ oxidation as effectively as did NH₄Cl in the fine sandy loam (82–89%), but less effectively in the sandy clay loam (52–66%). Acetylene (5 mol mol^-1 in soil headspace) had a strong (76–100%) inhibitory effect on CH₄ consumption in both soils. The phosphoroamide (urease inhibitor) N-(n-butyl) thiophosphoric triamide (NBPT) showed strong inhibition of CH₄ consumption at 25 g g^-1 soil in the fine sandy loam (83%) in the sandy clay loam (60%), but NH ₄ ⁺ oxidation inhibition was weak in both soils (13–17%). The discovery that the urease inhibitor NBPT inhibits CH₄ oxidation was unexpected, and the mechanism involved is unknown.     

Persistence of effects of nitrification inhibitors added to soils

Abstract

The persistence of the effects of four nitrification inhibitors (2‐ethynylpyridine, nitrapyrin, etridiazole, 3‐methylpyrazole‐l‐carboxamide) on nitrification in soil was assessed by measuring the ability of two soils to nitrify NH₄ ⁺ [added as (NH₄)₂SO₄] after they had been treated with 5 μg inhibitor g^‐1 soil and incubated at 10, 20, or 30°C for 0, 21, 42, 84, 126, or 168 days. The soils used differed markedly in organic‐matter content (1.2 and 4.2% organic C). The data obtained showed that the persistence of the effects of the inhibitors studied decreased markedly with increase in soil temperature from 10 to 30°C and that, whereas the initial inhibitory effects of the test compounds on nitrification were greatest with the soil having the lower organic‐matter content, the persistence of their effects at 20 or 30°C was greatest with the soil having the higher organic‐matter content. The inhibitory effects of 2‐ethynylpyridine and etridiazole on nitrification were considerably more persistent than those of nitrapyrin or 3‐methylpyrazole‐l‐carboxamide and were significant even after incubation of inhibitor‐treated soil at 20°C for 168 days.     

Stimulation of heterotrophic nitrification and N2O production,inhibition of autotrophic nitrification in soil by adding readily degradable carbon

Journal of Soils and Sediments - This study aimed to test the hypothesis that readily degradable Carbon (C) has contrasting effect on soil N autotrophic and heterotrophic nitrification, can...     

Mineralization and nitrification in three Malaysian forest soils

Examination of three forest soils from Malaysia using the soil incubation technique suggests that nitrification was not inhibited in these oligotrophic soils. Nitrification rates were between 40 and 750 ngN produced g^?1 dry weight soil day^?1 of incubation. Addition of phenolic metabolites (tannic acid) and leaf filtrates from hill and lowland forest litter did not significantly inhibit nitrification. Addition of sucrose (1% w/w carbon source) decreased nitrification but not ammonification.     

Evaluation of nitrification inhibitors for use under tropical conditions

Abstract

Six simple chemical compounds were evaluated for inhibition of nitrification. The compounds of o‐nitroaniline, m‐nitroaniline, o‐nitrophenol, benzotriazole, and 2,4,6‐trinitrophenol were compared with dicyandiamide by adding 10 μg inhibitor/g of a sandy loam soil, incubating for 30 days at 30°C and analyzing for NH⁺ ₄, NO^‐ ₂, and NO^‐ ₃ formation. The results indicated that benzotriazole, o‐nitrophenol, and m‐nitroaniline were very effective nitrification inhibitors.     

Effects of nitrification inhibitors on denitrification of nitrate in soil

Summary The influence of 28 nitrification inhibitors on denitrification of nitrate in soil was studied by determining the effects of different amounts of each inhibitor on the amounts of nitrate lost and the amounts of nitrite, N₂O and N₂ produced when soil samples were incubated anaerobically after treatment with nitrate or with nitrate and mannitol. The inhibitors used included nitrapyrin (N-Serve), etridiazole (Dwell), potassium azide, 2-amino-4-chloro-6-methylpyrimidine (AM), sulfathiazole (ST), 4-amino-1,2,4-triazole(ATC),2,4-diamino-6-trichloromethyl-s-triazine (CL-1580), potassium ethylxanthate, guanylthiourea (ASU), 4-nitrobenzotrichloride, 4-mesylbenzotrichloride, sodium thiocarbonate (STC), phenylmercuric acetate (PMA), and dicyandiamide (DCD).Only one of the nitrification inhibitors studied (potassium azide) retarded denitrification when applied at the rate of 10 g g^–1 soil, and only two (potassium azide and 2,4-diamino-6-trichloromethyl-s-triazine) inhibited denitrification when applied at the rate of 50 g g^–1 soil. The other inhibitors either had no appreciable effect on denitrification, or enhanced denitrification, when applied at the rate of 10 or 50 g g^–1 soil, enhancement being most marked with 3-mercapto-1,2,4-triazole. Seven of the inhibitors (potassium azide, sulfathiazole, potassium ethylxanthate, sodium isopropylxanthate, 4-nitrobenzotrichloride, sodium thiocarbonate, and phenylmercuric acetate) retarded denitrification when applied at the rate of 50 g g^–1 soil to soil that had been amended with mannitol to promote microbial activity.Reports that nitrapyrin (N-Serve) and etridiazole (Dwell) inhibit denitrification when applied at rates as low as 0.5 g g^–1 soil could not be confirmed. No inhibition of denitrification was observed when these compounds were applied at the rate of 10 g g^–1 soil, and enhancement of denitrification was observed when they were applied at the rate of 50 or 100 g g^–1 soil.     

Effects of nitrification inhibitors on germination of various seeds in soil

Summary The effects of 19 nitrificiation inhibitors on germination of seeds in soil were investigated. The nitrification inhibitors tested were sodium azide, potassium azide, potassium ethyl xanthate, nitrapyrin (N-Serve), etridiazole (Dwell), 3-mercapto-1,2,4-triazole (MT), 2-amino-4-chloro-6-methylpyrimidine (AM), 2,4-diamino-6-trichloromethyl-s-triazine, 2-mercaptobenzothiazole (MBT), 4-amino-1,2,4-triazole (ATC), sodium thiocarbonate (STC), guanylthiourea (ASU), thiourea (TU), dicyandiamide (DCD), sulfathiazole (STC), phenylacetylene, 2-ethynyl-pyridine, 3-methylpyrazole-l-carboxamide (MPC), and ammonium thiosulfate (ATS). Germination tests were performed with seeds of alfalfa (Medicago sativa L.), wheat (Triticum aestivum L.), rye (Secale cereale L.), barley (Hordeum vulgare L.), sorghum [Sorghum bicolor (L.) Moench], oats (Avena sativa L.), and corn (Zea mays). Only 2 of the 19 nitrification inhibitors studied (potassium azide and sodium azide) reduced germination of the seeds tested when applied at the rate of 12.5 g g^–1. The other inhibitors studied had no effect on the germination of wheat, alfalfa, barley, corn, oat, rye, or sorghum seeds when they were applied at the rate of 125 g g^–1 soil, and most of them had no effect on seed germination when applied at the rate of 625 g g^–1 soil.     

Effects of nitrification inhibitors on transformations of urea nitrogen in soils

The effects of three patented nitrification inhibitors on transformations of urea N in soils were studied by determining the effects of these compounds (10 μg/g of soil) on urea hydrolysis, ammonia volatilization. and production of ammonium, nitrite, and nitrate in soils incubated under aerobic conditions (30°C, 60% WHC) after treatment with urea (400 μg of urea N/g of soil). The inhibitors used (N-Serve, ATC, and CL-1580) had little, if any, effect on urea hydrolysis, but they retarded nitrification of the ammonium formed by urea hydrolysis and increased gaseous loss of urea N as ammonia. They also decreased the amount of (urea + exchangeable ammonium + nitrite + nitrate) — N found in urea-treated soils after various times.Two of the soils used accumulated substantial amounts of nitrite(> 160 μg of nitrite N/g of soil) when incubated under aerobic conditions after treatment with urea. Addition of nitrification inhibitors to these soils eliminated or substantially reduced nitrite accumulation and greatly retarded nitrate formation, but had little, if any, effect on the recovery of urea N as (urea + exchangeable ammonium + nitrite + nitrate + ammonia) — N after various times. This finding and other observations reported indicate that the “nitrogen deficits” observed in studies of urea N transformations in soils may not largely be due to gaseous loss of urea N through chemodenitrification and are at least partly due to volatilization and fixation of the ammonium formed by urea hydrolysis in soils. The work reported also indicates that N-Serve and other nitrification inhibitors may prove useful for reduction of the nitrite toxicity problems associated with the use of urea as a fertilizer but that application of such inhibitors in conjunction with fertilizer urea, when surface applied, may promote gaseous loss of urea N as ammonia.     

The effect of three fungicides on nitrification and ammonification in soil

Three fungicides, Captan, Thiram and Verdasan were added at varying concentrations to soil amended with ammonium sulphate, and their effect upon nitrification and ammonification was studied over 28 days. Two general effects of addition of fungicides on nitrification were apparent. At very low concentrations all three fungicides stimulated or did not affect this process. The stimulation was most marked after treatment with Thiram at 10 μg a.i./g soil. At higher concentrations the fungicides led to a progressive decrease in nitrate production. The concentration at which nitrification was inhibited was for Verdasan 10 μg, Thiram 100 μg and Captan above 250 μg a.i./g soil.At low concentration all three fungicides did not greatly affect ammonification. At increasing concentrations, however, there was a marked increase of NH⁺₄-N, compared with the controls. The lowest rates of application of the three fungicides resulted in most nitrification and least ammonification. The results are discussed in relation to the differential effects of the fungicides on the soil microbial population.     

Release of mineral N from soils: Influence of inhibitors of nitrification

Nitrification inhibitors (N-Serve, ATC, and CS₂) were added to soils without N fertilizers. While the amount of nitrification of NH₄⁺-N was reduced, so was the amount of ammonification of soil N. This effect was greater with ATC and CS₂ than with N-Serve. In three field experiments, the application in the fall of ATC at 22 kg ha^?1 mixed into the soil reduced the loss of soil mineral N in early spring. Apparently, the inhibition suppressed both ammonification and nitrification of soil N during the winter, and consequently there was less NO^?₃ in soil when the wet period occurred in the spring.     

Differential inhibition of human platelet aggregation by selected Allium thiosulfinates

Thiosulfinates (TSs) have been implicated as a principle source of the antiplatelet property of raw onion and garlic juice. The in vitro responses of human platelets to dosages of four TSs were measured using whole blood aggregometry and compared by regression analysis. Of the compounds evaluated, methyl methane-TS (MMTS), propyl propane-TS (PPTS), and 2-propenyl 2-propene-TS (allicin) are present in freshly cut Allium vegetables, whereas ethyl ethane-TS (EETS) has not been detected. All TSs were synthesized using a model reaction system. PPTS and allicin had the strongest antiplatelet activity at 0.4 mM, inhibiting aggregation by 90 and 89%, respectively. At the same concentration, EETS and MMTS were significantly weaker, inhibiting 74 and 26%, respectively. Combinations of TSs were not additive in their inhibition of aggregation, indicating that the antiplatelet potential of Allium extracts cannot be easily predicted by quantifying organosulfur components. EETS, PPTS, and allicin were significantly more potent platelet inhibitors than aspirin at nearly equivalent concentrations.     

The effect of biochar loading rates on soil fertility,soil biomass,potential nitrification,and soil community metabolic profiles in three different soils

Purpose

Biochar is increasingly being used as a soil amendment to both increase soil carbon storage and improve soil chemical and biological properties. To better understand the shorter-term (10 months) impacts of biochar on selected soil parameters and biological process in three different textured soils, a wide range of loading rates was applied.

Materials and methods

Biochar derived from eucalypt green waste was mixed at 0, 2.5, 5, 10 % (wt/wt) with a reactive black clay loam (BCL), a non-reactive red loam (RL) and a brown sandy loam (BSL) and placed in pots exposed to the natural elements. After 10 months of incubation, analysis was performed to determine the impacts of the biochar rates on the different soil types. Also, microbial biomass was estimated by the total viable counts (TVC) and DNA extraction. Moreover, potential nitrification rate and community metabolic profiles were assayed to evaluate microbial function and biological process in biochar-amended soils.

Results and discussion

The results showed that biochar additions had a significant impact on NH₄ and NO₃, total C and N, pH, EC, and soil moisture content in both a soil type and loading-dependent manner. In the heavier and reactive BCL, no significant impact was observed on the available P and K levels, or the total exchangeable base cations (TEB) and CEC. However, in the other lighter soils, biochar addition had a significant effect on the exchangeable Al, Ca, Mg, and Na levels and CEC. There was a relatively limited effect on microbial biomass in amended soils; however, biochar additions and its interactions with different soils reduced the potential nitrification at the higher biochar rate in the two lighter soils. Community metabolic profile results showed that the effect of biochar on carbon substrate utilization was both soil type and loading dependent. The BCL and BSL showed reduced rates of substrate utilization as biochar loading levels increased while the opposite occurred for the RL.

Conclusions

This research shows that biochar can improve soil carbon levels and raise pH but varies with soil type. High biochar loading rates may also influence nitrification and the function and activity of microbial community in lighter soils.