Journal of Environmental Quality 32:423-431 (2003)
© 2003 American Society of Agronomy, Crop Science Society of America, and Soil Science Society of America
TECHNICAL REPORTS
Atmospheric Pollutants and Trace Gases
Nitrous Oxide, Nitric Oxide, and Nitrogen Dioxide Fluxes from Soils after Manure and Urea Application
Hiroko Akiyama* and
Haruo Tsuruta
National Institute for Agro-Environmental Sciences (NIAES), 3-1-1 Kannondai, Tsukuba, Ibaraki 305-8604, Japan
* Corresponding author (ahiroko{at}affrc.go.jp)
Received for publication June 6, 2002.
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ABSTRACT
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Nitrous oxide is a greenhouse gas, and NO and NO2 play a key role in atmospheric chemistry. Nitrous oxide, NO, and NO2 fluxes from fertilized soils were measured six times per day by an automated flux monitoring system for one year, beginning on 21 May 1998. Pac choi (Brassica spp.) was cultivated for two months, and the plots were left fallow the remainder of the year. Two types of manure, poultry manure (PM) and swine manure (SM), and a chemical fertilizer, urea, were applied to the soil. The total amount of nitrogen applied in each case was 15 g N m-2. The total fluxes from PM, SM, and urea for the year were 184, 61.3, and 44.8 mg N m-2 for N2O, respectively; 9.95, 16.6, and 148 mg N m-2 for NO, respectively; and -6.21, -7.23, and -7.84 mg N m-2 for NO2, respectively. A negative correlation was found between the NO flux and the NO concentration of the chamber air just after the chamber was closed, when a flux from the atmosphere to soil was observed for 10 months. The mean gross NO production, the NO uptake rate constant, and the apparent compensation point for this period were 0.79 to 0.95 µg N m-2 h-1, 120 to 128 L m-2 h-1, and 5.65 to 7.35 ppbv, respectively.
Abbreviations: PM, poultry manure SM, swine manure WFPS, water-filled pore space
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INTRODUCTION
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NITROUS OXIDE (N2O) is a greenhouse gas and also participates in the destruction of the stratospheric ozone. Nitric oxide (NO) is a precursor of tropospheric ozone, which is a greenhouse gas formed by photochemical reaction. Nitrogen dioxide (NO2) is derived from NO by photochemical reaction, and is a precursor of nitric acid, which is a major component of acid rain. Nitric oxide and NO2 play a key role in atmospheric chemistry (Williams et al., 1992). It is important to measure those three gases simultaneously to understand the whole picture of nitrogen exchange between the soil and the atmosphere.
Agricultural soil is a major source of N2O, accounting for approximately 6.2 Tg N2O-N yr-1, which is 35% of the global annual emission (Kroeze et al., 1999), and it is also considered a major source of NO, accounting for approximately 5.5 Tg NO-N yr-1 (Davidson and Kingerlee, 1997). Nitrous oxide and NO are produced in soils by the microbial processes of nitrification and denitrification after the application of nitrogen fertilizer or organic matter to the field. Nitrification is an aerobic process, and denitrification is an anaerobic process (Davidson, 1991; Granli and Bockman, 1994).
Input of organic matter to the agricultural field is one important N2O source. After organic matter is added to soil, the C source for denitrification is increased, microbial activity is enhanced, O2 is consumed, and anaerobic microsites can develop (Granli and Bockman, 1994). Considerable N2O emissions from animal waste have been estimated. Bouwman et al. (1995) estimated the global N2O emission from animal waste to be 1.0 Tg N2O-N yr-1, based on an estimated production of 100 Tg animal waste N yr-1 and a 1% N2O emission rate from animal waste. They claim that their estimate is conservative because it is based on only a few measurements of urine or manure-treated soils and animals in stables. Input of organic waste to the agricultural soil would also be a NO source. However, there are few reports of NO emission from soils to which manure has been applied (Paul et al., 1993; Harrison et al., 1995).
The purpose of this study was to investigate N2O, NO, and NO2 fluxes from an Andosol field after the application of various manures.
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MATERIALS AND METHODS
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Field Management
The experiment was performed in the lysimeter fields at Tsukuba, Japan. Each lysimeter had an area of 9 m2 (3 x 3 m) and a depth of 1.2 m. The soil was an Andisol, which is formed by the weathering of volcanic ash under well-drained conditions, and which covers about 50% of the total area of upland fields in Japan. It is characterized by a low bulk density (Classification Committee of Cultivated Soils, 1996) and is favorable for maintaining aerobic conditions. The properties of Andisols have been previously described by Akiyama et al. (2000).
The three treatments, prepared in duplicate for this experiment, were as follows:
- Poultry manure (PM): the poultry manure was dried and granulated but did not contain bedding. It was obtained from a commercial poultry farm.
- Swine manure (SM): the swine manure did not contain straw and was obtained from the Ibaraki Prefecture swine farming experimental station. It was made by using a composting machine with automated mixing and aeration. After a 2-wk composting process, the manure was dried and powdered.
- Urea: A chemical N fertilizer was used for comparison with the manure treatments.
Pac choi was grown from 2 June to 14 July 1998 (48 d). The manure or fertilizer application date was 2 June 1998. Equal amounts of total nitrogen (15 g N m-2) were applied in each of the three treatments. The manure or fertilizer was broadcast and then incorporated. The properties of the manures are shown in Table 1. No vegetation remained on the field after the harvest.
Measurements
Nitrous oxide, NO, and NO2 fluxes were measured six times per day by with an automated flux monitoring system (Akiyama et al., 2000) for a year from 21 May 1998 to 20 May 1999. The system used six closed polycarbonate chambers with a cross-sectional area of 8100 cm2 and a height of 45 cm. For the flux measurement, the lid of each chamber was closed automatically for 30 min, and the air inside was drawn to the analysis room through a 10-m-long Teflon tube. Four air samples were taken every 8.5 min during closing time.
The gas sample was analyzed immediately after the sampling. The concentration of N2O was determined with a gas chromatograph (GC) equipped with an electron capture detector (ECD) (GC-14B; Shimadzu Corp., Kyoto, Japan). The concentrations of NO and NO2 were determined with a chemiluminescence NOx analyzer (Model 17; Thermo Environmental Instruments, Franklin, MA). The NO2 concentration was determined as: [NO2] = [NOx] - [NO].
The soil mineral nitrogen from the 0- to 5-cm depth was analyzed. For this analysis, 15-g samples of fresh soil were extracted with 100 mL of KCl solution (100 g KCl L-1). Nitrate was analyzed by the coppercadmium reduction method, and NH+4 was analyzed by the indophenol blue method in a continuous flow analyzer (TRRACS; Bran+Luebbe, Nordersterdt, Germany). The volumetric water content was measured twice a day at 2, 5, 10, and 15-cm depths with time domain reflectometry (TDR) moisture sensors (CS615; Campbell Scientific Instruments, Logan, UT), and the absolute value of the soil moisture content was determined with a calibration curve for Andisols (Hatano et al., 1995). The soil temperature at 2-, 5-, and 10-cm depths and the air temperature were also monitored every 10 min.
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RESULTS AND DISCUSSION
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Nitrous Oxide, Nitric Oxide, and Nitrogen Dioxide Fluxes
Nitrous oxide peaks (Fig. 1)
were observed for 2 weeks to 1 mo, and NO peaks (Fig. 2)
were observed for 1 to 2 weeks, just after the manure or fertilizer application.

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Fig. 1. Seasonal variation in the N2O flux (daily mean) from soil after the application of manure or urea. The treatments were poultry manure (PM; broken line), swine manure (SM; solid line), and urea (dotted line). F = fertilizer or manure application, H = harvest.
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Fig. 2. Seasonal variation in the NO flux (daily mean) from soil after the application of manure or urea. The treatments were poultry manure (PM; broken line), swine manure (SM; solid line), and urea (dotted line). F = fertilizer or manure application, H = harvest.
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During the cultivation period, the total N2O emissions from PM- and SM-amended plots were 592 and 163%, respectively, of that from urea-amended plots (Table 2a). During the cultivation period, the total NO emissions from PM- and SM-amended plots were 8 and 11%, respectively, of that from urea-amended plots. Nitrous oxide emission from the PM plot during the cultivation period was significantly higher than that from the SM plot, although the N contents of the applied manures and the soil mineral N of those plots were the same. Nitrous oxide and NO emissions for the period of noncultivation did not differ significantly between the plots (Table 2b).
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Table 2. Total N2O, NO, and NO2 fluxes during (a) the period of cultivation of pac choi, (b) the nonvegetation periods, and (c) the entire year (means ± SD). The treatments were poultry manure (PM), swine manure (SM), and urea.
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From the urea plot, the total NO emission during the cultivation period was significantly higher (p < 0.01) than the total N2O emission (Table 2a). We also found previously dominant NO emission from a urea plot in an Andisol field (Akiyama et al., 2000; Akiyama and Tsuruta, 2002). On the other hand, from the PM plot, the total NO emission during the cultivation period was significantly lower (p < 0.05) than the total N2O emission (Table 2a). This may be because the application of manure caused anaerobic microsites to develop in the soil with the result that denitrification was enhanced (Granli and Bockman, 1994). In a laboratory study, the NO to N2O emission ratio was greater than unity under nitrifying conditions and less than unity under denitrifying conditions (Anderson and Levine, 1986). During the same period, the soil mineral nitrogen content suggested that nitrification had occurred after manure application (Fig. 3) . Nitrous oxide emissions from nitrification can also increase as aeration becomes restricted (Granli and Bockman, 1994). Nitrification and denitrification probably take place simultaneously. Although the contribution of nitrification and denitrification was not determined in this experiment, the model to simulate the contribution of nitrification and denitrification for NO and N2O emissions was developed (Parton et al., 2001).

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Fig. 3. (a) Soil NO-3 and (b) NH+4 during the cultivation period. The treatments were poultry manure (PM; triangle), swine manure (SM; square), and urea (circle). F = fertilizer or manure application, H = harvest, DS = dry soil.
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Nitrogen dioxide was absorbed by all of the plots (Fig. 4
, Table 2); however, the absorption rate of 6.2 to 7.8 mg N m-2 yr-1 was much smaller than that reported previously by Akiyama and Tsuruta (2002), who estimated an absorption rate of 32.5 mg N m-2 yr-1. A negative correlation was found between NO2 flux and NO2 concentration just after the chamber was closed (Fig. 5b)
. No difference in the NO2 absorption rate was observed among the three plots, and no clear seasonal pattern in the NO2 flux was observed. No relationships between the NO2 flux and the water-filled pore space (WFPS) or temperature were observed. Nitrogen dioxide uptake has also been observed by Akiyama and Tsuruta (2002), Yamulki et al. (1997), Skiba et al. (1992), and Hirose and Tsuruta (1996). It has been reported that NO2 deposition to the soil is modulated by physicochemical factors rather than by biological factors (Baumgartner et al., 1992; Gasche and Papen, 1999), whereas N2O and NO fluxes are highly affected by biological factors (Williams et al., 1992). A series of chemical reactions, in which emitted NO is oxidized to NO2, followed by NO2 deposition onto the soil and leaf surfaces, is thought to occur in both agricultural and nonagricultural ecosystems. Such deposition processes must be taken into account to assess properly the effect of NO emissions from the soil (Williams et al., 1992).

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Fig. 4. Seasonal variation in the NO2 flux (daily mean) from soil after the application of manure or urea. The treatments were poultry manure (PM; broken line), swine manure (SM; solid line), and urea (dotted line). F = fertilizer or manure application, H = harvest.
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Fig. 5. Representative example of the relationship between (a) the NO-N flux and NO-N concentration and (b) the NO2N flux and NO2N concentration just after the chamber was closed. Measurements were made between 21 June 1998 and 30 Apr. 1999. The treatment was poultry manure (PM). Solid line: regression (a) y = -0.149x + 0.974, R2 = 0.62, p < 0.01; (b) y = -0.29x + 0.087, R2 = 0.560, p < 0.01.
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Note that NO2 fluxes reported here were measured by the closed chamber method. Thus, the calculated NO2 fluxes were highly dependent on the concentration in the chamber air just after the chamber was closed, and NO2 deposition fluxes may thus have been underestimated.
Apparent Nitric Oxide Compensation Point
The net flux of NO between soil and atmosphere is the result of simultaneous production and uptake processes:
where J = net NO flux between soil and atmosphere; P = NO production rate; k = uptake rate constant; m = NO mixing ratio; and mc = NO compensation point. The apparent NO compensation point is defined as NO mixing ratio in ambient air, at which the net flux between soil and atmosphere is zero (Slemr and Seiler, 1991; Gasche and Papen, 1999, Conrad et al., 1991; Godde and Conrad, 1998; Bollmann et al., 1999). The partial pressure of NO in the soil reaches a steady state value when the rates of NO formation and its simultaneous destruction are equal. Deposition of NO occurs if the mixing ratio of NO in the air is higher than this steady state partial pressure in the soil, and emission occurs if it is lower. When NO flux rates (y axis) are plotted against NO mixing ratios (x axis), the apparent compensation concentration (mc) is given by the intercept of the regression line with the x axis (Fig. 5a). From the intercept of the regression line with the y axis, the gross NO production (P) can be calculated, whereas the slope of the regression line represents the NO uptake rate constant (k) (Slemr and Seiler, 1991; Gasche and Papen, 1999; Conrad et al., 1991).
In this experiment, NO uptake was observed, except for the first month after manure or fertilizer application. For the period from 21 June 1998 to 30 Apr. 1999, when the flux from atmosphere to soil was observed, a negative correlation was found between NO flux and the NO concentration in the chamber air measured just after the chamber was closed (Fig. 5a). The mean gross NO production, the NO uptake rate constant, and the apparent compensation did not differ among the plots (Table 3). Their values during this period were similar to those that we reported previously (Akiyama and Tsuruta, 2002). Slemr and Seiler (1991) reported that the compensation point depends on environmental variables, such soil temperature and water content. In this experiment, however, no clear seasonal pattern of those values and no relationships between them and WFPS or temperature were observed. Although soil was a net sink for NO from 21 June 1998 to 30 Apr. 1999, it was a net source after organic matter or fertilizer application, from 21 May to 20 June 1998, as well as for the year (Table 2).
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Table 3. Apparent NO compensation points (mc), NO uptake rate constant (k), and mean gross NO production rate (P) from 21 June 1998 to 30 Apr. 1999.
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Most of the reported values of NO compensation points from field measurements were of the same order of magnitude as the ambient NO mixing ratio, ranging from 0 to 75 ppbv (Slemr and Seiler, 1991; Kim et al., 1994; Johansson and Granat, 1984; Gasche and Papen, 1999). The apparent compensation point in this experiment (5.657.35 ppbv) was also within this range (Table 3).
Nitric oxide consumption is considered to be a biological process, and Johansson and Galbally (1984) showed that no uptake of NO occurs in autoclaved soil. Nitric oxide consumption has been reported both in heterotrophic microorganisms (Dunfield and Knowles, 1998; Conrad, 1996) and autotrophic nitrite oxidizers (Conrad, 1996).
Diel Pattern of Nitrous Oxide and Nitric Oxide Fluxes
A diel pattern was observed in N2O and NO fluxes in all of the plots on most days during the peak emission period, which was similar to the pattern of air temperature fluctuation (Fig. 6) . No clear time difference between the diel peak of N2O and NO fluxes was observed. The diel pattern in the N2O and NO fluxes after the peak emission period was not as clear as peak emission period. No diel pattern in the NO2 flux was observed during the entire observation period (data not shown). We obtained similar results in our previous experiments (Akiyama et al., 2000; Akiyama and Tsuruta, 2002).

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Fig. 6. (a) Air temperature (solid line) and rainfall (vertical bars) from 7 to 13 June 1998. Representative example of diel variations during peak emission period in (b) N2O and (c) NO fluxes. Treatment was urea.
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Figure 6 shows a representative example of diel pattern in N2O and NO fluxes during peak emission period from the urea plot. In addition to temperature, rainfall events and soil mineral N content affected N2O and NO fluxes. A small rainfall of 0.5 mm on 9 June did not affect either N2O or NO. In contrast, when a rainfall of 27.5 mm was observed on 10 June, the NO flux decreased but the N2O flux was not affected. After the rainfall of 10 June, N2O emission gradually decreased. Nitric oxide emission on 11 June increased but was lower than that before the rainfall and then decreased on 12 June. The soil mineral N content from the 0- to 5-cm depth was also decreased after 10 June (Fig. 3). Nitrous oxide and NO emission did not respond to the temperature or a rainfall of 9 mm on 13 June, when the mineral N content of surface soil was low. Nitrous oxide and NO emissions generally increase with soil mineral N content (Granli and Bockman, 1994; Davidson and Verchot, 2000; Skiba et al., 1997; Skiba and Smith, 2000). Nitrous oxide and NO emissions generally increase with temperature (Skiba et al., 1997; Granli and Bockman, 1994; Martin et al., 1998), and the diel variation can be partly explained by variations in temperature (Granli and Bockman, 1994). Rainfall events also affect N2O and NO emissions by increasing WFPS, which controls NO to N2O emission ratio (Davidson, 1991; Davidson and Verchot, 2000; Parton et al., 2001). The large increase in NO fluxes following rainfall events when the soil had been dry before the rainfall events was also reported (Martin et al., 1998; Yienger and Levy, 1995; Skiba et al., 1997), although it was not observed in this experiment.
Soil Mineral Nitrogen
Soil NH+4 increased just after manure or fertilizer application in all of the plots, and NH+4 decreased and NO-3 increased during the peak N2O and NO emission periods (Fig. 3). This indicates that nitrification contributed to N2O and NO production. The NO to N2O emission ratio (5.17) suggests that nitrification was the main source of production in the urea plot (Table 2), because the NO to N2O emission ratio is greater than unity under nitrifying conditions and less than unity under denitrifying conditions, according to a laboratory study (Anderson and Levine, 1986). Nitrification and denitrification probably occur simultaneously, and the application of manure would develop anaerobic microsites in the soil, which would enhance denitrification (Granli and Bockman, 1994).
The soil mineral nitrogen content at the 0- to 5-cm depth decreased after 10 June, possibly because of NO-3 leaching by rainfall. Total rainfall from 10 to 14 June was 61 mm. The enhancement of NO fluxes also stopped at that time, although N2O emission from the PM and SM plots continued. This could be because rainfall stimulated N2O emission but not NO emission. High WFPS could also help create anaerobic conditions around the manure and stimulate N2O production. Hosen et al. (2000) described a model that NO emission decreased dramatically but N2O emission was not affected, when the N2O and NO production site became deeper. This could also explain why the N2O emission period was longer than that for NO.
Relationship between the Flux Ratio of Nitric Oxide to Nitrous Oxide and Water-Filled Pore Space
In the PM and urea plots, a negative correlation was found between water-filled pore space (WFPS) at the 5-cm depth (Fig. 7)
and the flux ratio of NO-N to N2O-N during the peak emission period (from 715 June 1998) (Fig. 8a,c)
. Similar results were obtained in our previous experiments (Akiyama et al., 2000; Akiyama and Tsuruta, 2002). Davidson (1991) described a model according to which NO emission is expected to be higher than N2O emission when WFPS is lower (WFPS = 2050%), and N2O emission is expected to be higher than NO emission when WFPS is higher (WFPS > 60%). Our experimental result, which was obtained in a light-textured Andisol, was consistent with this model. Davidson and Verchot (2000) and Parton et al. (2001) also reported a negative relationship between WFPS and the flux ratio of NO-N to N2O-N.

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Fig. 7. Seasonal variations in water-filled pore space (WFPS) at 2 cm (broken line; daily mean), 5 cm (solid line; daily mean), and rainfall (vertical bars; daily mean).
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Fig. 8. Relationships between the flux ratio of NO-N to N2O-N and water-filled pore space (WFPS) during the peak emission period (from 715 June 1998) (daily mean). The treatments were (a) poultry manure (PM), (b) swine manure (SM), and (c) urea. Solid line: regression (a) y = -0.0046x + 0.238, R2 = 0.453, p < 0.05; (c) y = -0.659x + 33.4, R2 = 0.441, p < 0.05.
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After the peak emission period, however, no correlation was seen between the flux ratio and WFPS in any treatment group (data not shown). In the SM plot, no correlation was observed during any period (Fig. 8b). In addition, no correlation was seen between the NO2 flux and WFPS during any period (data not shown).
Relationship between Soil Temperature and Nitrous Oxide and Nitric Oxide Fluxes
A clear seasonal temperature effect on N2O and NO fluxes was not observed. We found no correlation between soil and air temperatures (Fig. 9)
and N2O, NO, and NO2 fluxes (Fig. 1, 2, and 4), although a relationship was reported previously (Akiyama and Tsuruta, 2002). This kind of correlation can be seen only when other factors such as WFPS and mineral nitrogen content, which also affect N2O and NO emissions, are relatively constant.

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Fig. 9. Seasonal variation in soil temperature at 5 cm (broken line; daily mean) and air temperature (solid line; daily mean).
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Plant Nitrogen Uptake Rate
Nitrogen recovered by pac choi was 54.5 to 66.3% of applied N (Table 4), and total N2O-N, NO-N, and NO2N fluxes for the year were only 0.47 to 1.23% of applied N (Table 2c). Thus about 45 to 32% of applied N was not recovered in this experiment. Note that the plant N uptake from soil organic matter mineralization was not taken into account and net N uptake from applied N would be much lower than this calculation. A large part of applied N was lost possibly by NO-3 leaching. A 3-yr monitoring from 1997 to 1999 in a nearby Andisol field showed that the amount of rainfall was much greater than evapotranspiration, and downward water flow ranged from 227 to 762 mm yr-1 (Hasegawa and Eguchi, 2002). Ammonia volatilization would be another reason for N loss. Bouwman et al. (1997) estimated that average NH3 losses from urea and animal waste applications were about 15 and 20%, respectively, of applied N in the temperate zone. Ammonia loss from fertilizer is affected by the type of fertilizer, soil pH and application method (Bouwman et al., 1997). In this experiment, both urea and manure were incorporated, and soil pH was 5.9; thus, NH3 loss would be lower than the average emission rate. A part of applied N would also be lost through N2 emission by denitrification. Del Grosso et al. (2000) described a model according to which N2 to N2O ratio was less than unity, when WFPS was lower than 80%. Water-filled pore space was lower than 80% throughout the monitoring period (Fig. 7); thus, N2 emission would be a small part of N loss.
The N uptake rate was lowest (p < 0.05) in the SM plot (Table 4), and it would be necessary to apply chemical fertilizer along with SM to improve the quality of the harvest. In that case, the emissions would be comparable with or even higher than those of the other two plots, although the sum of N2O-N and NO-N emissions from the SM plot was the lowest among the three treatments in this experiment (Table 2).
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CONCLUSIONS
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Nitrous oxide, NO, and NO2 fluxes from an Andisol field after the application of poultry and swine manures and urea were investigated. Our study showed that manure is an important source of N2O and NO emissions. Especially, N2O emission from poultry manure was significantly higher than that from urea. Although soil to which fertilizer or manure had been applied was a net source of NO, NO uptake was not observed except for the first month after the manure or fertilizer application. This shows that Andisols can be an important sink for NO, especially in nonagricultural lands. Soil was a net sink for NO2, and manure or fertilizer application did not affect its uptake.
Nitrous oxide and NO emissions from an Andisol amended with urea were 0.19 and 0.99% of applied N, respectively (Table 2). Nitrous oxide and NO emissions from the Japanese Andisol field amended with chemical fertilizer ranged from 0.06 to 0.28%, and from 0.026 to 1.19% of applied N, respectively (Akiyama et al., 2000; Akiyama and Tsuruta, 2002; Hou and Tsuruta, 2002; Yan et al., 2001). In contrast, N2O and NO emissions from a Costa Rican Andisol amended with chemical fertilizer were higher than those from the Japanese Andisol, and ranged from 0.5 to 5%, and from 0.6 to 5.7% of applied N, respectively (Weitz et al., 1999; Weitz et al., 2001; Veldkamp and Keller, 1997). It should be noted that all of the emission data from Japanese Andisols were obtained within Tsukuba area, although Andisols cover about 50% of total area of upland fields in Japan. Soil temperatures after the fertilization were similar in the Japanese site and in the Costa Rican site. The difference of N2O emission rates may be partly explained by WFPS (3565% in Japan vs. 5098% in Costa Rica). The difference of NO emissions, however, cannot be explained by WFPS. More study is needed to investigate the differences.
It is hard to investigate the effect of manure on the atmospheric environment because a wide variety of manure is used, and also because the chemical composition of manure is varied. Thus, the effect of manure application on N2O and NO emissions from agricultural fields has not been investigated to the same degree as that of chemical fertilizer application. More study is needed to determine the environmental effect of organic matter added to agricultural fields.
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